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Sample records for fluorescence hplc dad

  1. Determination of 4-ethylphenol and 4-ethylguaiacol in wines by LC-MS-MS and HPLC-DAD-fluorescence.

    Science.gov (United States)

    Caboni, Pierluigi; Sarais, Giorgia; Cabras, Marco; Angioni, Alberto

    2007-09-05

    Volatile phenols produced by Brettanomyces dekkera have been associate with off-flavors of wines. A versatile liquid chromatography-tandem mass spectrometry together with an HPLC-DAD-fluorescence methods were developed for the quantitation of two phenols, 4-ethylphenol (4EP) and 4-ethylguaiacol (4EG), in red and white wines. For LC-MS-MS analysis, fortified wines were directly injected after a dilution with methanol, and levels of phenols were measured by monitoring the multiple reaction (MRM) transitions of precursor ions mass charge (m/z) 121 --> 106 for 4EP and (m/z) 151 --> 136 for 4EG. Qualitative and quantitative confirmation data were acquired simultaneously by monitoring alternative MRM transitions following an external standard method. Calibration was linear over a working range of 10 and 5000 microg/L. Limit of determination (LOD) and limit of quantification (LOQ) were 10 and 50 microg/L for both 4EG and 4EP. HPLC analysis phenols were separated with a gradient system of acetonitrile-water and detected using a diode array detector (DAD) at 280 nm, and for the fluorescence analysis, excitation and emission wavelengths of 260 and 305 nm were used. Quantitative analysis of 4EP and 4EG was performed by the standard addition method to avoid matrix interferences. Calibration was linear over a concentration range from 10 to 5000 microg/L for HPLC-DAD, from 1 to 10,000 microg/L for 4EP, and from 10 to 10,000 for 4EG for fluorescence analysis. LOD and LOQ for the DAD analysis were 10 and 50 microg/L for both 4EG and 4EP. For fluorescence analysis, LOD and LOQ were 1 and 5 microg/L for 4EP and 10 and 50, respectively, for 4EG. The proposed methods can be easily used for the qualitative and quantitative determination of 4EP and 4EG in wines affected by microbial contamination with yeasts of the Brettanomyces genus.

  2. Rapid screening and identification of compounds with DNA-binding activity from Folium Citri Reticulatae using on-line HPLC-DAD-MS(n) coupled with a post column fluorescence detection system.

    Science.gov (United States)

    Fu, Qingrong; Zhang, Cangman; Lin, Zongtao; Sun, Hongyang; Liang, Yi; Jiang, Haixiu; Song, Zhiling; Wang, Hong; Chen, Shizhong

    2016-02-01

    To study the interactions between natural compounds and deoxyribonucleic acid (DNA), a method has been established combining a high-performance liquid chromatography-diode array detector-multi-stage mass spectrometer with a fluorescence detector (HPLC-DAD-MS(n)-FLD). The FLD was used to monitor fluorescence intensity of the ethidium bromide-DNA (EB-DNA) complex when a compound separated by HPLC was introduced. This novel method was used to simultaneously obtain the HPLC fingerprint, UV spectra, MS(n) fragments and DNA-binding activity profile of various components in Folium Citri Reticulatae. As a result, 35 compounds were identified, of which 25 were found in the extract of Folium Citri Reticulatae for the first time, and 33 compounds showed DNA-binding activities, with the most active being feruloylhexaric and p-coumaroylhexaric acids. In addition, the precision, stability and reproducibility of this method were validated by two positive controls, quercetin and hesperidin. This new on-line method is accurate, precise and reliable for further high-throughput screening of DNA-binding compounds from food samples and other complex matrices.

  3. HPLC-DAD determination of imidacloprid in onion

    OpenAIRE

    Mandić Aljoša; Lazić Sanja; Inđić Dušanka

    2003-01-01

    Imidacloprid is an insecticide most commonly used on vegetables, potato sugar beet, fruit, cereal, maize and rice. Imidacloprid residue has been determined in spiked onion and in onion samples. Sample preparation consisted of dichlormethane extraction of imidacloprid from onion, followed by purification of the obtained extract on a LC-Florisil disposable cartridge. The HPLC-DAD method bas been developed on reversed-phase for separation of imidacloprid with a mixture of 0.01 M phosphate buffer...

  4. Rapid analysis of components in Rhizoma Anemarrhenae by HPLC-DAD-MS and HPLC-DAD-TOFMS

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    A global quality control method based on high performance liquid chromatography(HPLC)coupled with diode array detection(DAD),single quadrupole mass spectrometry(MS)and time-of-flight mass spectrometry(TOFMS)was developed for simultaneous determination of seven major components(mangiferin,neomangiferin,timosaponin E1,timosaponin E,timosaponin BⅡ,timosaponin BⅢ,and timosaponin AⅢ)and identification of most components in extracts of Rhizoma Anemarrhenae(RA).HPLC analysis was performed on an Agilent SB-C18 colu...

  5. Application of SPE-HPLC-DAD and SPE-TLC-DAD to the determination of pesticides in real water samples.

    Science.gov (United States)

    Tuzimski, Tomasz

    2008-10-01

    Planar chromatography with diode array scanning (TLC-DAD) and high-performance chromatography with diode array detection (HPLC-DAD) were used to screen water samples for pesticides. Pesticides were enriched from lake water samples by SPE on C18/SDB-1, C18, C18 Polar Plus and cyanopropyl (CN) cartridges. The recovery rates were high for all extraction materials except for all pesticides on CN cartridges, for which the values were lower. SPE was used not only for preconcentration of analytes but also for their fractionation. The analytes were eluted first with methanol and then with dichloromethane. Methanol eluates were analysed by HPLC-DAD, the dichloromethane eluates with TLC-DAD. The method was validated for precision, repeatability and accuracy. The calibration plots were linear between 0.1 and 50.0 microg/mL for all pesticides, the correlation coefficients, r, were between 0.9992 and 1.000 as determined by HPLC-DAD. In the TLC experiments, the best fit for the calibration lines was found when the calibration data were analysed using a second-degree polynomial regression. Calibration plots lay between 0.1 and 17 microg/spot for all pesticides, the correlation coefficients, r, were between 0.9974 and 0.9997 determined by TLC-DAD. The LOD was between 0.04 and 0.65 microg/spot (TLC-DAD) and between 0.02 and 3.68 microg/mL (HPLC-DAD).

  6. Simultaneous determination of 11 antibiotics and their main metabolites from four different groups by reversed-phase high-performance liquid chromatography-diode array-fluorescence (HPLC-DAD-FLD) in human urine samples.

    Science.gov (United States)

    Fernandez-Torres, R; Consentino, M Olías; Lopez, M A Bello; Mochon, M Callejon

    2010-05-15

    A new, accurate and sensitive reversed-phase high-performance liquid chromatography (RP-HPLC) as analytical method for the quantitative determination of 11 antibiotics (drugs) and the main metabolites of five of them present in human urine has been worked out, optimized and validated. The analytes belong to four different groups of antibiotics (sulfonamides, tetracyclines, penicillins and anphenicols). The analyzed compounds were sulfadiazine (SDI) and its N(4)-acetylsulfadiazine (NDI) metabolite, sulfamethazine (SMZ) and its N(4)-acetylsulfamethazine (NMZ), sulfamerazine (SMR) and its N(4)-acetylsulfamerazine (NMR), sulfamethoxazole (SMX), trimetroprim (TMP), amoxicillin (AMX) and its main metabolite amoxicilloic acid (AMA), ampicillin (AMP) and its main metabolite ampicilloic acid (APA), chloramphenicol (CLF), thiamphenicol (TIF), oxytetracycline (OXT) and chlortetracycline (CLT). For HPLC analysis, diode array (DAD) and fluorescence (FLD) detectors were used. The separation of the analyzed compounds was conducted by means of a Phenomenex Gemini C(18) (150mm x 4.6mm I.D., particle size 5microm) analytical column with LiChroCART LiChrospher C(18) (4mm x 4mm, particle size 5microm) guard column. Analyzed drugs were determined within 34min using formic acid 0.1% in water and acetonitrile in gradient elution mode as mobile phase. A linear response was observed for all compounds in the range of concentration studied. Two procedures were optimized for sample preparation: a direct treatment with methanol and acetonitrile and a solid phase extraction procedure using Bond Elut Plexa columns. The method was applied to the determination of the analytes in human urine from volunteers under treatment with different pharmaceutical formulations. This method can be successfully applied to routine determination of all these drugs in human urine samples.

  7. HPLC-DAD and HPLC-ESI-MS analyses of Tiliae flos and its preparations.

    Science.gov (United States)

    Karioti, A; Chiarabini, L; Alachkar, A; Fawaz Chehna, M; Vincieri, F F; Bilia, A R

    2014-11-01

    In the present study extensive HPLC-DAD, HPLC-ESI-MS and NMR analyses were undertaken in the aqueous preparations (decoctions, infusions) and tinctures of Tilia platyphyllos Scop inflorescences. The aim of this work was to examine in depth the qualitative and quantitative profile of the investigated preparations, which find until today wide applications in pharmaceutical and cosmetic industry, and to propose a validated method for their quality control. An HPLC-DAD-ESI-MS method was developed and optimised for the quantitative determination of the constituents. Marker constituents of Tiliae flos are the flavonoids, while the volatile content is also used for the quality control. However, the analyses of the non-volatile fraction gave complex chromatographic fingerprints containing simple phenolics and low molecular weight procyanidins. The use of different HPLC columns permitted a good separation of the constituents and enabled their quantitation, while HPLC-MS analyses permitted the detection of procyanidin oligomers. Overall, 31 constituents were detected and identified. Extensive preparative chromatographic investigations and 2D-NMR analyses allowed the characterisation of procyanidins as epicatechin derivatives. Finally, the HPLC method was validated and complied with ICH guidelines. This is the first report of detailed analysis of the chemical composition of Tiliae flos.

  8. Forensic intoxication with clobazam: HPLC/DAD/MSD analysis.

    Science.gov (United States)

    Proença, Paula; Teixeira, Helena; Pinheiro, João; Marques, Estela P; Vieira, Duarte Nuno

    2004-07-16

    Clobazam (Castillium, Urbanil), a benzodiazepine often used as an anxiolytic and in the treatment of epilepsy, is considered a relatively safe drug. The authors present a fatal case with a 49-year-old female, found dead at home. She had been undergoing psychiatric treatment and was a chronic alcoholic. The autopsy findings were unremarkable, except for multivisceral congestion, steatosis and a small piece of a plastic blister pack in the stomach. Bronchopneumonia, bronchitis and bronchiolitis were also diagnosed. Anhigh-performance liquid chromatography (HPLC)/diode array detector (DAD)/mass spectrometry detection (MSD) with electrospray method was developed in order to detect, confirm and quantify clobazam in the post-mortem samples. In the chromatographic separation, a reversed-phase column C18 (2.1 x 150 mm, 3.5 microm) was used with a mobile phase of methanol and water, at a 0.25 ml/min flow rate. Carbonate buffer (pH 10.5) and 20 microl of prazepam (100 microg/ml) as internal standard were added to the samples. A simple and reliable liquid-liquid extraction method for the determination of clobazam in post-mortem samples was described. Calibration curves for clobazam were performed in blood, achieving linearity between 0.01 and 10 microg/ml and a detection limit of 1.0 ng/ml. The clobazam concentration found in post-mortem blood was 3.9 microg/ml, higher than the reported therapeutic concentration (0.1-0.4 microg/ml). The simultaneous acquisition by photodiode array detection and mass spectrometry detection results allowed benzodiazepines to be identified with sufficient certainty. An examination of all the available information suggested that death resulted from respiratory depression due to clobazam toxicity.

  9. ANALYSIS OF THREE FLAVONOIDS INSCUTELLARIA BAICALENSIS GEORGI BY HPLC-DAD-MS%HPLC-DAD-MS法定量分析黄芩中3种黄酮类化合物

    Institute of Scientific and Technical Information of China (English)

    徐杨璐; 赵胜男; 申川川; 李守拙

    2014-01-01

    Objective:To compare the determination of three lfavonoids inScutellaria baicalensis Georgi by high performance liquid chromatography with diode array detection and mass spectrometer (HPLC-DAD-MS) and high performance liquid chromatography with diode array detection (HPLC-DAD).Methods: HPLC-DAD-MS and HPLC-DAD were respectively applied to determineBaicalin,Wogonoside and Wogonin inScutellaria baicalensis Georgi. Results:The content ofBaicalin, Wogonoside, Wogonin by HPLC-DAD-MS were 6.61%, 73.98% and 10.90% less than HPLC-DAD. Among them, the difference ofWogonoside content was fairly larger.Conclusions: Compared with HPLC-DAD, HPLC-DAD-MS is more accurate, simple and convenient. So HPLC-DAD-MS can be used for the determination of Baicalin, Wogonoside, WogonininScutellaria baicalensis Georgi.%目的::分析比较高效液相色谱-二极管阵列检测器-质谱(HPLC-DAD-MS)法和高效液相色谱-二极管阵列检测器(HPLC-DAD)法检测黄芩中3种黄酮类化合物的结果。方法:分别采用HPLC-DAD-MS法和HPLC-DAD法检测黄芩中3种黄酮类化合物黄芩苷、汉黄芩苷、汉黄芩素的含量。结果:HPLC-DAD-MS法测得黄芩苷、汉黄芩苷、汉黄芩素的含量分别比HPLC-DAD法平均少6.61%、73.98%和10.90%,其中汉黄芩苷的含量相差较大。结论:与HPLC-DAD法相比,HPLC-DAD-MS法含量测定结果更准确,且操作简便、重现性好,可用于黄芩中黄芩苷、汉黄芩苷、汉黄芩素的含量测定。

  10. Screening of Satureja subspicata Vis. Honey by HPLC-DAD, GC-FID/MS and UV/VIS: Prephenate Derivatives as Biomarkers

    National Research Council Canada - National Science Library

    Jerković, Igor; Kranjac, Marina; Marijanović, Zvonimir; Zekić, Marina; Radonić, Ani; Tuberoso, Carlo Ignazio Giovanni

    2016-01-01

    ...%. Bioprospecting of the samples was performed by HPLC-DAD, GC-FID/MS, and UV/VIS. Prephenate derivatives were shown to be dominant by the HPLC-DAD analysis, particularly phenylalanine (167.8 mg/kg...

  11. HPLC-DAD-MS identification of bioactive secondary metabolites from Ferula communis roots.

    Science.gov (United States)

    Arnoldi, Lolita; Ballero, Mauro; Fuzzati, Nicola; Maxia, Andrea; Mercalli, Enrico; Pagni, Luca

    2004-06-01

    A simple HPLC method was developed to distinguish between 'poisonous' and 'non-poisonous' chemotypes of Ferula communis. The method was performed on a C8 reverse phase analytical column using a binary eluent (aqueous TFA 0.01%-TFA 0.01% in acetonitrile) under gradient condition. The two chemotypes showed different fingerprints. The identification of five coumarins and eleven daucane derivatives by HPLC-diode array detection (HPLC-DAD) and HPLC-MS is described. A coumarin, not yet described, was detected.

  12. A Decomposition Model for HPLC-DAD Data Set and Its Solution by Particle Swarm Optimization

    Directory of Open Access Journals (Sweden)

    Lizhi Cui

    2014-01-01

    Full Text Available This paper proposes a separation method, based on the model of Generalized Reference Curve Measurement and the algorithm of Particle Swarm Optimization (GRCM-PSO, for the High Performance Liquid Chromatography with Diode Array Detection (HPLC-DAD data set. Firstly, initial parameters are generated to construct reference curves for the chromatogram peaks of the compounds based on its physical principle. Then, a General Reference Curve Measurement (GRCM model is designed to transform these parameters to scalar values, which indicate the fitness for all parameters. Thirdly, rough solutions are found by searching individual target for every parameter, and reinitialization only around these rough solutions is executed. Then, the Particle Swarm Optimization (PSO algorithm is adopted to obtain the optimal parameters by minimizing the fitness of these new parameters given by the GRCM model. Finally, spectra for the compounds are estimated based on the optimal parameters and the HPLC-DAD data set. Through simulations and experiments, following conclusions are drawn: (1 the GRCM-PSO method can separate the chromatogram peaks and spectra from the HPLC-DAD data set without knowing the number of the compounds in advance even when severe overlap and white noise exist; (2 the GRCM-PSO method is able to handle the real HPLC-DAD data set.

  13. Screening natural antioxidants in peanut shell using DPPH-HPLC-DAD-TOF/MS methods.

    Science.gov (United States)

    Qiu, Jiying; Chen, Leilei; Zhu, Qingjun; Wang, Daijie; Wang, Wenliang; Sun, Xin; Liu, Xiaoyong; Du, Fangling

    2012-12-15

    Peanut shell, a byproduct in oil production, is rich in natural antioxidants. Here, a rapid and efficient method using DPPH-HPLC-DAD-TOF/MS was used for the first time to screen antioxidants in peanut shell. The method is based on the hypothesis that upon reaction with 1, 1-diphenyl-2-picrylhydrazyl (DPPH), the peak areas of compounds with potential antioxidant activities in the HPLC chromatogram will be significantly reduced or disappeared, and the identity confirmation could be achieved by HPLC-DAD-TOF/MS technique. With this method, three compounds possessing potential antioxidant activities were found abundantly in the methanolic extract of peanut shell. They were identified as 5,7-dihydroxychromone, eriodictyol, and luteolin. The contents of these compounds were 0.59, 0.92, and 2.36 mg/g, respectively, and luteolin possessed the strongest radical scavenging capacity. DPPH-HPLC-DAD-TOF/MS assay facilitated rapid identification and determination of natural antioxidants in peanut shell, which may be helpful for value-added utilization of peanut processing byproducts.

  14. An HPLC-DAD method to quantification of main phenolic compounds from leaves of Cecropia Species

    OpenAIRE

    Geison M. Costa; Caroline F. Ortmann; Eloir P. Schenkel; Reginatto,Flávio H.

    2011-01-01

    An efficient and reproducible HPLC-DAD method was developed and validated for the simultaneous quantification of major compounds (chlorogenic acid, isoorientin, orientin and isovitexin) present in the leaves of two Cecropia species, C. glaziovii and C. pachystachya. From the leaves of C. glaziovii and C. pachystachya were isolated the C-glycosylflavones isoorientin and isovitexin and identified on both species chlorogenic acid (3-O-caffeoylquinic acid) and the O-glycosylflavonol isoquercitrin...

  15. A Decomposition Model for HPLC-DAD Data Set and Its Solution by Particle Swarm Optimization

    OpenAIRE

    Lizhi Cui; Zhihao Ling; Josiah Poon; Poon, Simon K.; Junbin Gao; Paul Kwan

    2014-01-01

    This paper proposes a separation method, based on the model of Generalized Reference Curve Measurement and the algorithm of Particle Swarm Optimization (GRCM-PSO), for the High Performance Liquid Chromatography with Diode Array Detection (HPLC-DAD) data set. Firstly, initial parameters are generated to construct reference curves for the chromatogram peaks of the compounds based on its physical principle. Then, a General Reference Curve Measurement (GRCM) model is designed to transform these p...

  16. Comparative evaluation of an ISO 3632 method and an HPLC-DAD method for safranal quantity determination in saffron.

    Science.gov (United States)

    García-Rodríguez, M Valle; López-Córcoles, Horacio; Alonso, Gonzalo L; Pappas, Christos S; Polissiou, Moschos G; Tarantilis, Petros A

    2017-04-15

    The aim of this work was a comparison of the ISO 3632 (2011) method and an HPLC-DAD method for safranal quantity determination in saffron. Samples from different origins were analysed by UV-vis according to ISO 3632 (2011) and by HPLC-DAD. Both methods were compared, and there was no correlation between the safranal content obtained by UV-vis and HPLC-DAD. An over-estimation in the UV-vis experiment was observed, which was related to the cis-crocetin esters content, as well as other compounds. The results demonstrated that there was no relationship between ISO quality categories and safranal content using HPLC-DAD. Therefore, HPLC-DAD might be preferable to UV-vis for determining the safranal content and the classification of saffron for commercial purposes. In addition, HPLC-DAD was adequate for determining the three foremost parameters that define the quality of saffron (crocetin esters, picrocrocin and safranal); therefore, this approach could be included in the ISO 3632 method (2011).

  17. Extraction, Separation, and Identification of Phenolic Compounds in Virgin Olive Oil by HPLC-DAD and HPLC-MS

    Science.gov (United States)

    Tasioula-Margari, Maria; Tsabolatidou, Eleftheria

    2015-01-01

    The aim of this study was to evaluate the recovery of individual phenolic compounds extracted from virgin olive oil (VOO), from different Greek olive varieties. Sufficient recoveries (90%) of all individual phenolic compounds were obtained using methanol as an extraction solvent, acetonitrile for residue solubilization, and two washing steps with hexane. Moreover, in order to elucidate structural characteristics of phenolic compounds in VOO, high performance liquid chromatography with a diode array detector (HPLC-DAD) at 280 and 340 nm and HPLC coupled to electrospray ionization mass spectrometry (HPLC-ESI-MS) in the negative-ion mode were performed. The most abundant phenolic compounds were oleuropein derivatives with m/z 319 and 377 and ligstroside derivatives with m/z 303, 361. Lignans, such as 1-acetoxypinoresinol and pinoresinol were also present in substantial quantities in the phenolic fraction. However, pinoresinol was co-eluted with dialdehydic form of ligstroside aglycone (DAFLA) and it was not possible to be quantified separately. The phenolic extracts, obtained from different VOO samples, yielded similar HPLC profiles. Differences, however, were observed in the last part of the chromatogram, corresponding to isomers of the aldehydic form of ligstroside aglycone. Oxidized phenolic products, originating from secoiridoids, were also detected. PMID:26783843

  18. Extraction, Separation, and Identification of Phenolic Compounds in Virgin Olive Oil by HPLC-DAD and HPLC-MS

    Directory of Open Access Journals (Sweden)

    Maria Tasioula-Margari

    2015-08-01

    Full Text Available The aim of this study was to evaluate the recovery of individual phenolic compounds extracted from virgin olive oil (VOO, from different Greek olive varieties. Sufficient recoveries (90% of all individual phenolic compounds were obtained using methanol as an extraction solvent, acetonitrile for residue solubilization, and two washing steps with hexane. Moreover, in order to elucidate structural characteristics of phenolic compounds in VOO, high performance liquid chromatography with a diode array detector (HPLC-DAD at 280 and 340 nm and HPLC coupled to electrospray ionization mass spectrometry (HPLC-ESI-MS in the negative-ion mode were performed. The most abundant phenolic compounds were oleuropein derivatives with m/z 319 and 377 and ligstroside derivatives with m/z 303, 361. Lignans, such as 1-acetoxypinoresinol and pinoresinol were also present in substantial quantities in the phenolic fraction. However, pinoresinol was co-eluted with dialdehydic form of ligstroside aglycone (DAFLA and it was not possible to be quantified separately. The phenolic extracts, obtained from different VOO samples, yielded similar HPLC profiles. Differences, however, were observed in the last part of the chromatogram, corresponding to isomers of the aldehydic form of ligstroside aglycone. Oxidized phenolic products, originating from secoiridoids, were also detected.

  19. Extraction, Separation, and Identification of Phenolic Compounds in Virgin Olive Oil by HPLC-DAD and HPLC-MS.

    Science.gov (United States)

    Tasioula-Margari, Maria; Tsabolatidou, Eleftheria

    2015-08-13

    The aim of this study was to evaluate the recovery of individual phenolic compounds extracted from virgin olive oil (VOO), from different Greek olive varieties. Sufficient recoveries (90%) of all individual phenolic compounds were obtained using methanol as an extraction solvent, acetonitrile for residue solubilization, and two washing steps with hexane. Moreover, in order to elucidate structural characteristics of phenolic compounds in VOO, high performance liquid chromatography with a diode array detector (HPLC-DAD) at 280 and 340 nm and HPLC coupled to electrospray ionization mass spectrometry (HPLC-ESI-MS) in the negative-ion mode were performed. The most abundant phenolic compounds were oleuropein derivatives with m/z 319 and 377 and ligstroside derivatives with m/z 303, 361. Lignans, such as 1-acetoxypinoresinol and pinoresinol were also present in substantial quantities in the phenolic fraction. However, pinoresinol was co-eluted with dialdehydic form of ligstroside aglycone (DAFLA) and it was not possible to be quantified separately. The phenolic extracts, obtained from different VOO samples, yielded similar HPLC profiles. Differences, however, were observed in the last part of the chromatogram, corresponding to isomers of the aldehydic form of ligstroside aglycone. Oxidized phenolic products, originating from secoiridoids, were also detected.

  20. Determination of acequinocyl and hydroxyacequinocyl on fruits and vegetables by HPLC-DAD.

    Science.gov (United States)

    Caboni, Pierluigi; Sarais, Giorgia; Melis, Marinella; Cabras, Marco; Cabras, Paolo

    2004-11-03

    A method for determining residues of the new reduced-risk pesticide acequinocyl and its deacetylated derivative hydroxyacequinocyl on fruits and vegetables (grapes, lemons, pears, and tomatoes) by HPLC is described. The pesticides were extracted from the fruits and vegetables with hexane and ethyl acetate solution (1:1, v/v), determined by HPLC-DAD at 250 nm and confirmed by LC/MS. No cleanup was necessary. This method is characterized by recoveries (0.01-4 mg/kg) > 77%, while the coefficient of variation was determined to be less than 11%. The limit of quantitation for both acequinocyl and hydroxyacequinocyl was 0.01 mg/kg for all matrixes.

  1. Analysis of bioactivities and chemical composition of Ziziphus joazeiro Mart. using HPLC-DAD.

    Science.gov (United States)

    Brito, Sharlene M O; Coutinho, Henrique D M; Talvani, Andre; Coronel, Cathia; Barbosa, Andreza G R; Vega, Celeste; Figueredo, Fernando G; Tintino, Saulo R; Lima, Luciene F; Boligon, Aline A; Athayde, Margareth L; Menezes, Irwin R A

    2015-11-01

    The aim of this study was to evaluate the chemical profile and antioxidant, antimicrobial and antiparasitic activities of the hydroalcoholic extract of the leaves of Ziziphus joazeiro Mart. (HELZJ). The antioxidant DPPH and FRAP assays and chemical profile were determined by colorimetric methods and HPLC/DAD. The antiparasitic, antibiotic and antibiotic-modifying activity were evaluated by microdilution assays. The HPLC-DAD assay showed the presence of mostly tannins and flavonoids, such as caffeic acid and quercetin. The levels of polyphenols and flavonoids were 183.136 mg/g extract and 7.37 mg/g extract, respectively. DPPH and FRAP showed low antioxidant activity for the extract. The antibacterial and antifungal activities were not of clinical relevance, showing MIC>1024 μg/mL. However, synergism was observed between HELZJ and the antibiotics amikacin and gentamicin, which resulted in decreased bacterial drug resistance. EHFZJ showed low toxicity in fibroblasts in vitro, while antiparasitic results against Trypnosoma cruzi, Leishmania braziliensis and Leishmania infantum were not clinically relevant. Thus, our results indicate that Z. joazeiro Mart. (HELZJ) could be a source of plant-derived natural products that could lead to the development of promising new antibiotic compounds for infectious diseases.

  2. Validation of AN Hplc-Dad Method for the Classification of Green Teas

    Science.gov (United States)

    Yu, Jingbo; Ye, Nengsheng; Gu, Xuexin; Liu, Ni

    A reversed phase high performance liquid chromatography (RP-HPLC) separation coupled with diode array detection (DAD) and electrospray ionization mass spectrometer (ESI/MS) was developed and optimized for the classification of green teas. Five catechins [epigallocatechin (EGC), epigallocatechin gallate (EGCG), epicatechin (EC), gallocatechin gallate (GCG), epicatechin gallate (ECG)] had been identified and quantified by the HPLC-DAD-ESI/MS/MS method. The limit of detection (LOD) of five catechins was within the range of 1.25-15 ng. All the analytes exhibited good linearity up to 2500 ng. These compounds were considered as chemical descriptors to define groups of green teas. Chemometric methods including principal component analysis (PCA) and hierarchical cluster analysis (HCA) were applied for the purpose. Twelve green tea samples originating from different regions were subjected to reveal the natural groups. The results showed that the analyzed green teas were differentiated mainly by provenance; HCA afforded an excellent performance in terms of recognition and prediction abilities. This method was accurate and reproducible, providing a potential approach for authentication of green teas.

  3. Methodological Establishment of HPLC-DAD for Determination of 10 Active Ingredients in Wuweizi

    Directory of Open Access Journals (Sweden)

    Feng YAO

    2015-09-01

    Full Text Available Objective: To establish high performance liquid chromatography-diode array detection (HPLC-DAD method for the determination of active ingredients of multiple lignans in Nanwuweizi (Kadsura longepedunculata and Beiwuweizi (Fructus Schisandrae Chinensis so as to set up the standard for the quality control of Wuweizi (Schisandra chinensis. Methods: Brava BDS C18 column (250 mm×4.6 mm, 5 μm was applied, with detection condition as follows: column temperature: 30℃; mobile phase: water (A-acetonitrile (B; gradient elution: 0.8 mL/min and detection wavelength: 225 nm.Results: Ten ingredients in Beiwuweizi and 5 in Nanwuweizi were completely separately within 75 min. Peak area was in favorable linear relationship with concentration. In addition, the recovery rate was 96.87-103.29% and relative standard deviation (RSD was 0.68-2.09%.Conclusion: HPLC-DAD is simple and reliable with favorable repeatability, so it can be used for the determination of multiple lignans in Wuweizi.

  4. Methodological Establishment of HPLC-DAD for Determination of 10 Active Ingredients in Wuweizi

    Institute of Scientific and Technical Information of China (English)

    YAO Feng; CHEN Da-shuai

    2015-01-01

    Objective: To establish high performance liquid chromatography-diode array detection (HPLC-DAD) method for the determination of active ingredients of multiple lignans in Nanwuweizi (Kadsura longepedunculata) and Beiwuweizi (Fructus Schisandrae Chinensis) so as to set up the standard for the quality control of Wuweizi (Schisandra chinensis). Methods: Brava BDS C18 column (250 mm×4.6 mm, 5 μm) was applied, with detection condition as follows: column temperature: 30℃; mobile phase: water (A)-acetonitrile (B); gradient elution: 0.8 mL/min and detection wavelength: 225 nm. Results: Ten ingredients in Beiwuweizi and 5 in Nanwuweizi were completely separately within 75 min. Peak area was in favorable linear relationship with concentration. In addition, the recovery rate was 96.87%-103.29% and relative standard deviation (RSD) was 0.68%-2.09%. Conclusion: HPLC-DAD is simple and reliable with favorable repeatability, so it can be used for the determination of multiple lignans in Wuweizi.

  5. Stability study of Prulifloxacin and Ulifloxacin in human plasma by HPLC-DAD.

    Science.gov (United States)

    Locatelli, Marcello; Cifelli, Roberta; Carlucci, Giuseppe; Romagnoli, Annalisa

    2016-01-01

    A new and specific HPLC-DAD method for the direct determination of Prulifloxacin and its active metabolite, Ulifloxacin, in human plasma has been developed. Plasma samples were analysed after a simple solid phase extraction (SPE) clean-up using a new HILIC stationary phase based high-performance liquid chromatography (HPLC) column and an ammonium acetate buffer (5 mM, pH 5.8)/acetonitrile (both with 1% Et(3)N, v/v) mobile phase in isocratic elution mode, with Danofloxacin as the internal standard. Detection was performed using DAD from 200 to 500 nm and quantitative analyses were carried out at 278 nm. The LOQ of the method was 1 μg/mL of the cited analytes and the calibration curve showed a good linearity up to 25 μg/mL. For both analytes the precision (RSD%) and the trueness (bias%) of the method fulfil with International Guidelines. The method was applied for stability studies, at three QC concentration levels, in human plasma samples stored at different temperature of + 25, + 4 and -20 °C in order to evaluate plasma stability profiles.

  6. 苦茶Camellia assamica var.kucha茶多酚的HPLC-DAD/MS分析%HPLC-DAD/MS Analysis of Tea Polyphenols in Camellia assamica var. kucha

    Institute of Scientific and Technical Information of China (English)

    王冬梅; 杜丽丽; 卢嘉丽; 石祥刚; 宋晓虹; 杨得坡; 叶创兴

    2006-01-01

    苦茶是一种特殊的茶组植物,它以1,3,7,9-四甲基尿酸为主要的嘌呤生物碱.本文采用高效液相色谱-二极管阵列检测/质谱联用(HPLC-DAD/MS)的方法对苦茶水提液中的茶多酚和嘌呤生物碱进行了定性分析,检测到3种嘌呤生物碱、7种儿茶素类化合物和2种非儿茶素类茶多酚(没食子酰奎宁酸酯和咖啡酰奎宁酸酯).同时与传统绿茶在儿茶素类化合物的组成和含量上进行了比较,结果显示,苦茶中总儿茶素含量(13.82%)远高于传统绿茶(7.37%),但各儿茶素类化合物的相对组成比在两种茶中极其相近,均以酯型儿茶素为主.

  7. Investigation of red natural dyes used in historical objects by HPLC-DAD-MS.

    Science.gov (United States)

    Karapanagiotis, Ioannis; Chryssoulakis, Yannis

    2006-01-01

    High performance liquid chromatography (HPLC) with UV-Vis Diode Array Detection (DAD) and electrospray mass spectrometric (ESI-MS) method was utilized for the identification of coloring components of madder, Armenian and Mexican cochineal, lac dye, brazilwood, safflower and dragon blood--probably the most important red natural dyestuffs found in objects of the cultural heritage. UV-Vis detection limits in the range of 0.2-0.6 ng for carminic acid, alizarin and purpurin were achieved using a gradient elution of H2O-0.01% TFA and CH3CN-0.01% TFA. ESI mass spectrometer was also used, as a supportive detection method to the standard DAD, for further analysis of the tested materials, with the ability to analyze dyestuffs as small as one milligram. The presence of madder was revealed in two historical (Hellenistic and Roman period) samples, found in the Mediterranean area, by identifying purpurin in both of them. Munjistin was also identified in one of the samples (Hellenistic period) while alizarin was not detected, raising questions regarding the exact madder type, utilized in the historical samples.

  8. Analysis of clobazam and its active metabolite norclobazam in plasma and serum using HPLC/DAD.

    Science.gov (United States)

    Akerman, K K

    1996-11-01

    This report describes a simple reversed-phase high-performance liquid chromatographic (HPLC) method with automated solid-phase extraction (SPE) for analysing clobazam and norclobazam concentrations in human serum or plasma. For the HPLC analysis the samples and standards are prepared with an ASPEC automatic sample preparer using 100-mg Bond-Elut C-18 solid-phase extraction columns. The HPLC method is an isocratic method with a mobile phase of acetonitrile:methanol:10 mmol l-1 dipotassium hydrogen phosphate, pH 3.7 (30:2:100), at a flow rate of 1.5 ml min-1. The benzodiazepines are detected with a diode array detector (DAD) at 240 nm and the peak purity analyses are performed at 210-365 nm. The recovery is over 97% for both analytes, and it is independent of the drug concentration. The intra-assay CVs vary between 0.7 and 2.2% and inter-assay CVs between 3.8 and 4.6% at therapeutic drug concentrations. The detection limit is 15 nmol l-1. The assay is linear from 30 to 20,000 nmol l-1 (clobazam) and from 170 to 105,000 nmol l-1 (norclobazam). This method leads to a very good separation of norclobazam from carbamazepine and phenytoin. None of the anti-epileptic or antidepressant drugs tested interfere with the assay.

  9. HPLC-DAD-ESI-MS/MS screening of bioactive components from Rhus coriaria L. (Sumac) fruits.

    Science.gov (United States)

    Abu-Reidah, Ibrahim M; Ali-Shtayeh, Mohammed S; Jamous, Rana M; Arráez-Román, David; Segura-Carretero, Antonio

    2015-01-01

    Rhus coriaria L. (sumac) is an important crop widely used in the Mediterranean basin as a food spice, and also in folk medicine, due to its health-promoting properties. Phytochemicals present in plant foods are in part responsible for these consequent health benefits. Nevertheless, detailed information on these bioactive compounds is still scarce. Therefore, the present work was aimed at investigating the phytochemical components of sumac fruit epicarp using HPLC-DAD-ESI-MS/MS in two different ionisation modes. The proposed method provided tentative identification of 211 phenolic and other phyto-constituents, most of which have not been described so far in R. coriaria fruits. More than 180 phytochemicals (tannins, (iso)flavonoids, terpenoids, etc.) are reported herein in sumac fruits for the first time. The obtained results highlight the importance of R. coriaria as a promising source of functional ingredients, and boost its potential use in the food and nutraceutical industries.

  10. An improvement of window factor analysis for resolution of noisy HPLC-DAD data

    Institute of Scientific and Technical Information of China (English)

    邵学广; 林祥钦; 邵利民; 李梅青

    2002-01-01

    Window factor analysis (WFA) is a powerful tool in analyzing evolutionary process. However, it was found that window factor analysis is much sensitive to the noise involved in original data matrix. An error analysis was done with the fact that the concentration profiles resolved by the conventional window factor analysis are easily distorted by the noise reserved by the abstract factor analysis (AFA), and a modified algorithm for window factor analysis was proposed. Both simulated and experimental HPLC-DAD data were investigated by the conventional and the improved methods. Results show that the improved method can yield less noise-distorted concentration profiles than the conventional method, and the ability for resolution of noisy data sets can be greatly enhanced.

  11. Chemical fingerprinting of Gardenia jasminoides Ellis by HPLC-DAD-ESI-MS combined with chemometrics methods.

    Science.gov (United States)

    Han, Yan; Wen, Jun; Zhou, Tingting; Fan, Guorong

    2015-12-01

    A fingerprint analysis method has been developed for characterization and discrimination of Gardenia jasminoides Ellis from different areas. The chemometrics methods including similarity evaluation, principal components analysis (PCA) and hierarchical clustering analysis (HCA) were introduced to identify more useful chemical markers for improving the quality control standard of dried ripe fruits of G. jasminoides Ellis. Then the selected chemical markers were analyzed by high performance liquid chromatography-diode array detection-electrospray ionization mass spectrometry (HPLC-DAD-ESI-MS) qualitatively and quantitatively. 23 characteristic peaks were assigned while 19 peaks of them were identified by comparing retention times, UV and MS spectra with authentic compounds or literature data. Moreover, 14 of them were determined quantitatively which could effectively evaluate the quality of G. jasminoides Ellis. This study was expected to provide comprehensive information for the quality evaluation of G. jasminoides Ellis, which would be a valuable reference for further study and development of this herb and related medicinal products.

  12. HPLC-DAD determination of mepivacaine in cerebrospinal fluid from a fatal case.

    Science.gov (United States)

    Nieddu, Maria; Boatto, Gianpiero; Serra, Domenico; Soro, Aldo; Lorenzoni, Salvatore; Lubinu, Francesco

    2007-09-01

    A fatal case involving mepivacaine-induced epidural anesthesia is described. The pathological findings were typical of cardiac shock from ischemic origin. Cerebrospinal fluid (CSF) was obtained several hours after death and mepivacaine was identified by gas chromatography-mass spectrometry (GC-MS). Its concentration was determined by high performance liquid chromatography with diode array detection (HPLC-DAD). Extraction from CSF was performed by deproteinization with acetonitrile. The mepivacaine concentration in the sample was 264 microg/mL. Concentrations of mepivacaine in CSF following epidural anesthesia are not reported in literature to our knowledge. This is the first reported case of death in which the mepivacaine concentration in CSF has been determined.

  13. Qualitative and quantitative analysis of chemical constituents of Centipeda minima by HPLC-QTOF-MS & HPLC-DAD.

    Science.gov (United States)

    Chan, Chi-On; Jin, Deng-Ping; Dong, Nai-Ping; Chen, Si-Bao; Mok, Daniel Kam Wah

    2016-06-05

    A high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (HPLC-QTOF-MS) method in both positive and negative ion modes was established to investigate the major constituents in the ethanolic extract of Centipeda minima (EBSC). Twelve common components including flavones and their glycosides, phenolic and polyphenolic acids, and sesquiterpene lactone were identified in ten batches of samples based on comparison with the retention time and accurate mass of external standards (mass accuracy within 3ppm) or the fragmentation patterns of tandem MS. Meanwhile, a simple, accurate and reliable HPLC-DAD method was also developed to determine the content of 10 chemical markers simultaneously. Results obtained from method validations including linearity, accuracy and precision showed that this new method is reliable and robust. Isochlorogenic acid A and brevilin A were found to be the most abundant in the ethanol extract of EBSC and could be served as markers for quality control of EBSC. Copyright © 2016 Elsevier B.V. All rights reserved.

  14. Determination of some psychotropic drugs in serum and saliva samples by HPLC-DAD and HPLC MS.

    Science.gov (United States)

    Petruczynik, A; Wróblewski, K; Szultka-Młyńska, M; Buszewski, B; Karakuła-Juchnowicz, H; Gajewski, J; Morylowska-Topolska, J; Waksmundzka-Hajnos, M

    2016-08-05

    A simple, rapid and sensitive HPLC-DAD method has been developed and validated for the simultaneous determination of seven psychotropic drugs (risperidone, citalopram, clozapine,quetiapine, levomepromazine, perazine and aripiprazole) in human serum or saliva samples. The chromatographic analyses were performed on a XSELECT CSH Phenyl-Hexyl column with a mobile phase containing methanol, acetate buffer at pH 3.5 and 0.025mL(-1) diethylamine. The influence of concentration of methanol in injection samples and injection volume on peak symmetry and system efficiency was examined.The full separation of all investigated drugs, good peaks' symmetry and simultaneously high systems efficiency were obtained in applied chromatographic system. The method is suitable for the analysis of investigated drugs in human plasma or saliva for psychiatric patients for control of pharmacotherapy, particularly in combination therapy. HPLC-MS was applied for verification of the presence of drugs and their metabolites in serum and saliva samples from patients.

  15. HPLC-DAD-MS analysis of dyes identified in textiles from Mount Athos.

    Science.gov (United States)

    Mantzouris, Dimitrios; Karapanagiotis, Ioannis; Valianou, Lemonia; Panayiotou, Costas

    2011-03-01

    Organic colorants contained in 30 textiles (16th to early 20th century) from the monastery of Simonos Petra (Mount Athos) have been investigated using high-performance liquid chromatography equipped with diode-array detection and mass spectrometry (HPLC-DAD-MS). The components of natural dyes identified in samples treated by the standard HCl dyestuff extraction method were: alizarin, apigenin, butein, carminic acid, chrysoeriol, dcII, dcIV, dcVII, ellagic acid, emodin, fisetin, flavokermesic acid, fustin, genistein, haematein derivative (Hae'), indigotin, indirubin, isoliquiritigenin, isorhamnetin, kaempferide, kaempferol, kermesic acid, luteolin, naringenin, purpurin, quercetin, rhamnazin, rhamnetin, sulfuretin, and type B and type C compounds (last two are markers for Caesalpinia trees). Early, semi-synthetic dyes, for example indigo carmine, fuchsin components, and rhodamine B were identified in objects dated late 19th to early 20th century. A dyestuff extraction method which involves use of TFA, instead of HCl, was applied to selected historical samples, showing that the mild method enables efficient extraction of weld (Reseda luteola L.) and dyer's broom (Genista tinctoria L.) glycosides. The marker compound (Hae') for logwood (Haematoxylum campechianum L.) identification after treatment with HCl was investigated by liquid chromatography coupled to mass spectrometry (LC-MS) in negative electrospray ionization (LC-MS-ESI(-)) mode. LC-MS in negative atmospheric pressure chemical ionization (LC-MS-APCI(-)) mode was used, probably for the first time, to investigate cochineal (Dactylopius coccus Costa) samples. Positive electrospray ionization (LC-MS-ESI(+)) mode was used for identification of fuchsin components. Detailed HPLC-DAD studies were performed on young fustic (Cotinus coggygria Scop.) and Persian berries (Rhamnus trees).

  16. HPLC-DAD-ESI-MS Analysis of Flavonoids from Leaves of Different Cultivars of Sweet Osmanthus

    Directory of Open Access Journals (Sweden)

    Yiguang Wang

    2016-09-01

    Full Text Available Osmanthus fragrans Lour. has traditionally been a popular ornamental plant in China. In this study, ethanol extracts of the leaves of four cultivar groups of O. fragrans were analyzed by high-performance liquid chromatography coupled with diode array detection (HPLC-DAD and high-performance liquid chromatography with electrospray ionization and mass spectrometry (HPLC-ESI-MS. The results suggest that variation in flavonoids among O. fragrans cultivars is quantitative, rather than qualitative. Fifteen components were detected and separated, among which, the structures of 11 flavonoids and two coumarins were identified or tentatively identified. According to principal component analysis (PCA and hierarchical cluster analysis (HCA based on the abundance of these components (expressed as rutin equivalents, 22 selected cultivars were classified into four clusters. The seven cultivars from Cluster III (‘Xiaoye Sugui’, ‘Boye Jingui’, ‘Wuyi Dangui’, ‘Yingye Dangui’, ‘Danzhuang’, ‘Foding Zhu’, and ‘Tianxiang Taige’, which are enriched in rutin and total flavonoids, and ‘Sijigui’ from Cluster II which contained the highest amounts of kaempferol glycosides and apigenin 7-O-glucoside, could be selected as potential pharmaceutical resources. However, the chemotaxonomy in this paper does not correlate with the distribution of the existing cultivar groups, demonstrating that the distribution of flavonoids in O. fragrans leaves does not provide an effective means of classification for O. fragrans cultivars based on flower color.

  17. Simultaneous determination of prenylflavonoid and hop bitter acid in beer lee by HPLC-DAD-MS.

    Science.gov (United States)

    Kao, T H; Wu, G Y

    2013-11-15

    An HPLC-DAD-MS method with high accuracy and precision was developed for determination of prenylflavonoids and hop bitter acids in beer lee, a by-product from beer brewing process. Four prenylflavonoids and nine hop bitter acids can be simultaneously separated in 29 min using a Thermo HyPURITY C18 column in combination with diode array dectector and mass spectrometer with HPLC solvent gradient system of phosphoric acid aqueous solution at pH 1.6 and acetonitrile at a flow rate of 1.5 mL/min and detection wavelength at 314 nm. Beer lee is found to contain isoxanthohumol (36.2 μg/g), xanthohumol (29.6 μg/g), 8-prenylnaringenin (7.84 μg/g), 6-prenylnaringenin (19.2 μg/g), cohumulone (44.7 μg/g), humulone (123 μg/g), adhumulone (21.8 μg/g), colupulone (44.2 μg/g), lupulone (33.2 μg/g), and adlupulone (5.76 μg/g).

  18. HPLC-DAD-ESI-MS Analysis of Flavonoids from Leaves of Different Cultivars of Sweet Osmanthus.

    Science.gov (United States)

    Wang, Yiguang; Fu, Jianxin; Zhang, Chao; Zhao, Hongbo

    2016-09-14

    Osmanthus fragrans Lour. has traditionally been a popular ornamental plant in China. In this study, ethanol extracts of the leaves of four cultivar groups of O. fragrans were analyzed by high-performance liquid chromatography coupled with diode array detection (HPLC-DAD) and high-performance liquid chromatography with electrospray ionization and mass spectrometry (HPLC-ESI-MS). The results suggest that variation in flavonoids among O. fragrans cultivars is quantitative, rather than qualitative. Fifteen components were detected and separated, among which, the structures of 11 flavonoids and two coumarins were identified or tentatively identified. According to principal component analysis (PCA) and hierarchical cluster analysis (HCA) based on the abundance of these components (expressed as rutin equivalents), 22 selected cultivars were classified into four clusters. The seven cultivars from Cluster III ('Xiaoye Sugui', 'Boye Jingui', 'Wuyi Dangui', 'Yingye Dangui', 'Danzhuang', 'Foding Zhu', and 'Tianxiang Taige'), which are enriched in rutin and total flavonoids, and 'Sijigui' from Cluster II which contained the highest amounts of kaempferol glycosides and apigenin 7-O-glucoside, could be selected as potential pharmaceutical resources. However, the chemotaxonomy in this paper does not correlate with the distribution of the existing cultivar groups, demonstrating that the distribution of flavonoids in O. fragrans leaves does not provide an effective means of classification for O. fragrans cultivars based on flower color.

  19. Antioxidant capacity and phenolic compounds of Lonicerae macranthoides by HPLC-DAD-QTOF-MS/MS.

    Science.gov (United States)

    Hu, Xin; Chen, Lin; Shi, Shuyun; Cai, Ping; Liang, Xuejuan; Zhang, Shuihan

    2016-05-30

    Lonicerae macranthoides with strong antioxidant activity is commonly used in traditional Chinese medicine and folk tea/beverage. However, detailed information about its antioxidant activity and bioactive compounds is limited. Then at first, we comparatively evaluated total phenolic content (TPC), total flavonoid content (TFC) and antioxidant activities of water extract, petroleum ether, ethyl acetate and n-butanol fractions of L. macranthoides. Ethyl acetate fraction exhibited the highest level of TPC (207.38 mg GAE/g DW), TFC (53.06 mg RE/g DW) and the best DPPH scavenge activity and reducing power. n-Butanol fraction showed the best ABTS(+) and O2(-) scavenging activities. Interestingly, water extract, ethyl acetate and n-butanol fractions showed stronger antioxidant activities than positive control, butylated hydroxytoluene (BHT). After that, thirty-one antioxidant phenolic compounds, including twenty-two phenolic acids and nine flavonoids, were screened by DPPH-HPLC experiment and then identified using HPLC-DAD-QTOF-MS/MS. It is noted that twenty-one compounds (1, 3-4, 6-17, 19, 23, 26, 28-29, and 31), as far as was known, were discovered from L. macranthoide for the first time, and eleven of them (3-4, 10-17, and 23) were reported in Lonicera species for the first time. Results indicated that L. macranthoides could serve as promising source of rich antioxidants in foods, beverages and medicines for health promotion.

  20. Improved quantification of pyrogenic carbon in soils and sediments by a HPLC-DAD method

    Science.gov (United States)

    Wiedemeier, D. B.; Hilf, M. D.; Smittenberg, R. H.; Schmidt, M. W. I.

    2012-04-01

    Fire-derived (pyrogenic) carbon (PyC) is produced by the incomplete combustion of biomass, for example during wildfires. It can persist in the environment for a long time due to its relative resistance against biological and chemical breakdown. Its accurate quantification in soils and sediments is of great interest because the slow turn-over of PyC has implications for the global carbon cycle and carbon budget calculations. Moreover, PyC in pedological and sedimentological records can be used to reconstruct wildfire history or to investigate historical periods like the industrialization. A whole suite of PyC quantification methods exists because PyC is not a defined chemical structure but rather a continuum of thermally altered biomass. The benzene polycarboxylic acids (BPCA) analysis is a molecular marker method that was shown to give conservative estimates of PyC quantity in soils. In addition, it yields qualitative information about the degree of aromaticity and condensation of PyC. The commonly used BPCA method consists in digesting samples with nitric acid that breaks down the PyC into a suite of BPCAs, which are cleaned, derivatized and finally analyzed by gas chromatography-flame ionization detection (GC-FID). Here, we present a modified BPCA method for soils and sediments that uses a high performance liquid chromatography system coupled to diode array detection (HPLC-DAD). We demonstrate that this method greatly enhances the reproducibility of PyC quantification in soil and sediment samples while significantly reducing analysis time. Moreover, much less sample material is needed for precise PyC quantification and we show that the HPLC-DAD method yields consistently higher PyC contents than the GC-FID method. Additionally, the modified method also facilitates δ13C and 14C measurements of the PyC fraction in these complex matrix samples. The isotopic information further improves the assessment of PyC budgets in the environment and the reconstruction of past

  1. Simultaneous determination of metronidazole and tinidazole in plasma by using HPLC-DAD coupled with second-order calibration

    Institute of Scientific and Technical Information of China (English)

    Li Qun Ouyang; Hai Long Wu; Ya Juan Liu; Jian Yue Wang; Yong Jie Yu; Hong Yan Zou; Ru Qin Yu

    2010-01-01

    A method using HPLC-DAD coupled with second-order calibration was developed to simultaneously determine metronidazole and tinidazole in plasma samples in this paper.The second-order calibration method based on APTLD(alternating penalty trilinear decomposition)algorithm was proposed to analyze the three-way HPLC-DAD data from both standard and prediction samples,which makes it possible that calibration can be performed even in the presence of unknown interferences with a simple and green chromatographic condition and short analysis time.The results showed that good recoveries were obtained although the chromatographic and spectral profiles of the analytes of interest as well as background were partially overlapped with each other in plasma samples.

  2. Comparative HPLC/ESI-MS and HPLC/DAD study of different populations of cultivated, wild and commercial Gentiana lutea L.

    Science.gov (United States)

    Mustafa, Ahmed M; Caprioli, Giovanni; Ricciutelli, Massimo; Maggi, Filippo; Marín, Rosa; Vittori, Sauro; Sagratini, Gianni

    2015-05-01

    The root of Gentiana lutea L., famous for its bitter properties, is often used in alcoholic bitter beverages, food products and traditional medicine to stimulate the appetite and improve digestion. This study presents a new, fast, and accurate HPLC method using HPLC/ESI-MS and HPLC/DAD for simultaneous analysis of iridoids (loganic acid), secoiridoids (gentiopicroside, sweroside, swertiamarin, amarogentin) and xanthones (isogentisin) in different populations of G.lutea L., cultivated in the Monti Sibillini National Park, obtained wild there, or purchased commercially. Comparison of HPLC/ESI-MS and HPLC/DAD indicated that HPLC/ESI-MS is more sensitive, reliable and selective. Analysis of twenty samples showed that gentiopicroside is the most dominant compound (1.85-3.97%), followed by loganic acid (0.11-1.30%), isogentisin (0.03-0.48%), sweroside (0.05-0.35%), swertiamarin (0.08-0.30%), and amarogentin (0.01-0.07%). The results confirmed the high quality of the G.lutea cultivated in the Monti Sibillini National Park.

  3. Study on chromatographic fingerprint of sarcandra glabra (Thunb.) by microwave-assisted extraction coupled to HPLC/DAD

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    Microwave-assisted extraction(MAE)was used for extraction of effective components of sarcandra glabra(Thunb.),and then chromatographic fingerprint of sarcandra glabra(Thunb.)was studied by high performance liquid chromatography/diode array detector(HPLC/DAD).The conditions of MAE were optimized by an orthogonal experiment,and then the authentication and validation of the chromatographic fingerprint were conducted.Nine peaks were identified as common peaks in the fingerprint chromatograms,and isofraxidin was...

  4. HPLC-DAD-ELSD Combined Pharmacodynamics and Serum Medicinal Chemistry for Quality Assessment of Huangqi Granule.

    Directory of Open Access Journals (Sweden)

    Huaguo Chen

    Full Text Available To more scientifically and reasonably control the quality of Huangqi Granules, preliminary studies on the pharmacodynamics and serum pharmacochemistry of this medicine were performed. DPPH and MTT experiments showed that water extracts of Huangqi Granules had good antioxidant activity and increased immunity. Timed blood samples collected 5 min, 15 min, and 30 min after oral administration of a set amount of Huangqi Granules were collected and tested using UPLC-ESI-MS/MS. As a result, calycosin-7-O-β-D-glucoside, ononin, calycosin, astragaloside IV, and formononetin were found to exist in rat blood after dosing, indicating that the five chemical compounds might have pharmacological activity, and based on this result, they were designated biomarkers for quality control of Huangqi Granules. Consequently, a simple, rapid and efficient method was developed in the present study for the simultaneous determination of the five characteristic compounds in Huangqi Granules using HPLC-DAD-ELSD.The separation was performed using an Agilent Hypersil ODS column (4.6 × 250 mm, 5 μm at 30 ℃. The mobile phase was composed of water (solvent A and acetonitrile (solvent B with a flow rate of 1 mL/min. The drift tube temperature of the ELSD system was set to 85 ℃, and the nitrogen pressure was 3.5 bar.All five characteristic compounds had good linear behavior with r2 values greater than 0.9972. The recoveries varied from 96.31% to 101.22%. Subsequently, the developed method was applied to evaluate the quality of Huangqi Granules from different batches, and hierarchical clustering analysis (HCA was used to analyze the classification of the samples based on the values of the five compounds.The established HPLC method combined with HCA proved to be effective to evaluate the quality of Huangqi Granules.

  5. A Validated HPLC-DAD Method for Simultaneous Determination of Etodolac and Pantoprazole in Rat Plasma

    Directory of Open Access Journals (Sweden)

    Ali S. Abdelhameed

    2014-01-01

    Full Text Available A simple, sensitive, and accurate HPLC-DAD method has been developed and validated for the simultaneous determination of pantoprazole and etodolac in rat plasma as a tool for therapeutic drug monitoring. Optimal chromatographic separation of the analytes was achieved on a Waters Symmetry C18 column using a mobile phase that consisted of phosphate buffer pH~4.0 as eluent A and acetonitrile as eluent B in a ratio of A : B, 55 : 45 v/v for 6 min, pumped isocratically at a flow rate of 0.8 mL min−1. The eluted analytes were monitored using photodiode array detector set to quantify samples at 254 nm. The method was linear with r2=0.9999 for PTZ and r2=0.9995 for ETD at a concentration range of 0.1–15 and 5–50 μgmL−1 for PTZ and ETD, respectively. The limits of detection were found to be 0.033 and 0.918 μgmL−1 for PTZ and ETD, respectively. The method was statistically validated for linearity, accuracy, precision, and selectivity following the International Conference for Harmonization (ICH guidelines. The reproducibility of the method was reliable with the intra- and interday precision (% RSD <7.76% for PTZ and <7.58 % for ETD.

  6. Quantititative determination of lycorine and galanthamine in Galanthus trojanus and G. cilicicus by HPLC-DAD.

    Science.gov (United States)

    Kaya, Gulen Irem; Polat, Derya Cicek; Sarikaya, Buket; Onur, Mustafa Ali; Somer, Nehir Unver

    2014-08-01

    Lycorine and galanthamine have various biological activities. A reliable HPLC method coupled with DAD detection was developed and validated for the determination of galanthamine and lycorine in Galanthus trojanus and G. cilicicus. A simple method for the extraction of the alkaloids in low-mass plant samples was employed utilizing columns pre-packed with diatomaceous earth (Extrelut). This method was applied to the aerial parts and bulbs of G. trojanus and G. cilicicus (Amaryllidaceae) collected during the flowering season. The chromatographic separation was performed using an isocratic system with a mobile phase of trifluoroacetic acid-water-acetonitrile (0.01:92.5:7.5) applied at a flow rate of 1 mL min(-1) and using a diode array detector. Validation procedures showed that the method was specific, accurate and precise. The highest amount of lycorine (0.012%) was detected in the bulbs of G. trojanus collected from Can (Canakkale), whereas the aerial parts of this species collected from Bayramiç (Canakkale) was not found to contain this alkaloid. In G. cilicicus samples, lycorine was only determined in the bulbs, giving yields of 0.004%; galanthamine yields were between 0.015-0.016%, but none of the G. trojanus samples contained this latter alkaloid.

  7. Determination of EDTA in feed and premix formulations by HPLC-DAD.

    Science.gov (United States)

    Chiumiento, Francesco; D'Aloise, Antonio; Marchegiani, Francesca; Melai, Valeria

    2015-05-15

    A simple analytical method for the quantitative determination of ethylenediaminetetraacetic acid (EDTA) in feed and premix formulations was developed and validated. The method involves an extraction with an acidic ferric chloride solution, to quantitatively convert EDTA species in the samples into the Fe(III)-EDTA complex, and its subsequent detection by Ion-Pair-Reversed Phase-High Performance Liquid Chromatography-Diode Array Detection (IP-RP-HPLC-DAD). A robust validation procedure was performed according to the Decision 2002/657/EC at concentrations ranging from 25 to 100 mg kg(-1) on sample. Good recoveries (85.6-92.8%) were obtained; repeatability of the method was in the range of 1.3-8.0%, with an intermediate precision ranging from 6.0% to 8.6%, both of them expressed as relative standard deviation (RSD). No interfering species hindered the straightforward detection of EDTA. Hence, the proposed method can be adopted for an effective and rapid routine analysis of products for livestock.

  8. An HPLC-DAD method to quantification of main phenolic compounds from leaves of Cecropia species

    Energy Technology Data Exchange (ETDEWEB)

    Costa, Geison M.; Ortmann, Caroline F.; Schenkel, Eloir P.; Reginatto, Flavio H., E-mail: freginatto@hotmail.co [Universidade Federal de Santa Catarina (UFSC), Florianopolis (Brazil). Centro de Ciencias da Saude. Dept. de Ciencias Farmaceuticas

    2011-07-01

    An efficient and reproducible HPLC-DAD method was developed and validated for the simultaneous quantification of major compounds (chlorogenic acid, isoorientin, orientin and isovitexin) present in the leaves of two Cecropia species, C. glaziovii and C. pachystachya. From the leaves of C. glaziovii and C. pachystachya were isolated the C-glycosylflavones isoorientin and isovitexin and identified on both species chlorogenic acid (3-O-caffeoylquinic acid) and the O-glycosylflavonol isoquercitrin. The C-glycosylflavone orientin was isolated only from C. pachystachya. Chlorogenic acid was the major compound in both species (11.1 mg g{sup -1} of extract of C. glaziovii and 27.2 mg g{sup -1} of extract of C. pachystachya) and for the flavonoids quantified, isovitexin was the main C-glycosylflavonoid for C. glaziovii (4.6 mg g{sup -1} of extract) and isoorientin the main one for C. pachystachya (17.3 mg g{sup -1} of extract). (author)

  9. Quality Evaluation and Regional Analysis of Psoraleae Fructus by HPLC-DAD-MS/MS plus Chemometrics

    Institute of Scientific and Technical Information of China (English)

    YAN Dong-mei; CHANG Yan-xu; KANG Li-yuan; GAO Xiu-mei

    2010-01-01

    Objective To evaluate the quality of Psoralea corylifolia collected from 12 provinces of China.Methods An HPLCDAD-MS/MS method was used to identify,determine,and estimate 14 representative bioactive compounds in P.corylifolia.Then on the basis of the content data,the chemometrics method was used to differentiate 20 samples from different regions.Results The quality of P.corylifolia from 12 different provinces of China was evaluated by this method.Though the samples showed similar profiles,content of the detected markers varied significantly in different regions and batches.According to the results of the hierarchical cluster analysis and principle component analysis,it can be concluded that the samples from different origins could be clustered reasonably into two groups,as well as successfully distinguished.Conclusion A simple and reliable new method which used HPLC-DAD-MS/MS and chemometrics has been developed to characterize,classify,and control the quality of P.corylifolia.

  10. Determination of relative response factors of the opium alkaloids with HPLC-DAD

    Directory of Open Access Journals (Sweden)

    Jelena Acevska

    2012-02-01

    Full Text Available In this work, a convenient method for determination of relative UV response factors (RRFs of morphine, codeine, thebaine, oripavine, noscapine and papaverine by high performance liquid chromatography (HPLC equipped with a diode array detector (DAD was presented. Pholcodine was selected as the reference compound for calculating the relative response factors of the alkaloids. The separation of all seven compounds was obtained with optimized gradient elution with high pH value of the mobile phase on a reversed phase column with bidentate C18-C18 bonding technology. The RRFs of the alkaloids were determinate by three different approaches: ‘regression analysis/mass concentration’, ‘regression analysis/molar concentration’and ‘detector sensitivity’ approaches. The ‘regression analysis/molar concentration’ approach gave the accurate approximation of the exact amount of the substance that enters in the detector and the statistically relevant calculation includes several points of different concentrations (at least five, which makes this approach most advantageous one. This method is suitable for quality assessment of the standardised opium dry extract, raw opium and standardised opium tincture by quantitative analysis of not only morphine and codeine as indicated in the respective European Pharmacopoeia monographs, but as well as the major impurities that originate from opium poppy Papaver somniferum L. (Papaveraceae.

  11. Development of complementary HPLC-DAD/APCI MS methods for chemical characterization of pharmaceutical packaging materials.

    Science.gov (United States)

    Petruševski, V; Jolevska, S T; Ribarska, J T; Chachorovska, M; Petkovska, A; Ugarković, S

    2016-05-30

    The chemical characterization of plastics for pharmaceutical packaging has been subject to ever increasing regulatory scrutiny, the reasons for which being: a) plastic additives and degradation products can be extremely hazardous to the patients' health (especially patients on chronic therapy) and b) they offer no therapeutic or formulatory benefit whatsoever. The last decade has seen the issuing of several books, monographs and guidelines dealing with extractables and leachables, however the amount of scientific work done so far is still fairly small (the majority of it performed by only a few research groups), with only a small number of methods published in the literature. This work focuses on developing a set of two complementary HPLC-DAD/APCI MS methods for simultaneous separation, detection, identification and quantification of a wide variety of packaging additives and degradants, the second method specifically targeting a group of compounds known as polymeric hindered amine light stabilizers (HALS), which are known to be notoriously difficult to separate and analyze with standard analytical techniques. The methods are capable of detecting plastic additives present in low ppb concentrations, from samples extracted in solvents with various polarities and pH values. Both methods were developed and optimized using system suitability mixtures comprised of 9 additives commonly encountered in plastic materials, and their practical applicability tested on a variety of extracts from low-density polyethylene (LDPE) and polypropylene (PP), where several additives were successfully separated, detected and identified.

  12. HPLC-DAD-ESI/MS(n) profiling of phenolic compounds from Lathyrus cicera L. seeds.

    Science.gov (United States)

    Ferreres, F; Magalhães, S C Q; Gil-Izquierdo, A; Valentão, P; Cabrita, A R J; Fonseca, A J M; Andrade, P B

    2017-01-01

    Lathyrus cicera L. seeds are of interest for food and feed purposes. Despite the recognized antioxidant activity of the seeds, arising from the phenolic fraction, their phenolic compounds have not been studied in depth yet. Therefore, to determine the phenolics profile of these seeds, a target analysis was performed using high-performance liquid chromatography coupled to photodiode-array detection and electrospray ionization/ion trap mass spectrometry (HPLC-DAD-ESI/MS(n)). Thirty-seven glycosylated flavonoids were identified for the first time in the seeds of this species and, according to their MS fragmentation, clustered in flavonol-3-O-di-/tri-glycosides-7-O-rhamnosides and other flavonol-glycosides, and flavonol-3-O-(cinnamoyl)glycoside-7-O-rhamnosides, flavonol-3-O-(dihydrophaseoyl, cinnamoyl)glycoside-7-O-rhamnosides and flavonol-3-O-(malonyl)glycoside-7-O-rhamnosides. Glycosides of kaempferol were the main flavonoids found (10 non-acylated and 21 acylated), followed by those of quercetin (3) and those of isorhamnetin, apigenin and luteolin (1). The most abundant flavonols were identified as kaempferol-3-O-(2-hexosyl)hexoside-7-O-rhamnosides. The methodology used allowed to increase the knowledge on a relevant phytochemical class of seeds from L. cicera.

  13. Analysis of benzo[c]phenanthridine alkaloids in Eschscholtzia californica cell culture using HPLC-DAD and HPLC-ESI-MS/MS.

    Science.gov (United States)

    Son, Seok Young; Rhee, Hong Soon; Lee, Min Woo; Park, Jong Moon

    2014-01-01

    Effective HPLC-DAD and HPLC-ESI-MS/MS methods have been developed for the analysis of eight benzo[c]phenanthridine alkaloids (sanguinarine, chelirubine, macarpine, chelerythrine, dihydrosanguinarine, dihydrochelirubine, dihydromacarpine and dihydrochelerythrine), which are important metabolites in Eschscholtzia californica cell culture. By adopting a ternary gradient pump system, the dihydro-form alkaloids hardly separable from each other could be successfully separated, and all the target alkaloids could be simultaneously quantified with the LOD values of 0.01-0.79 μg/mL and the LOQ values of 0.03-3.59 μg/mL. This HPLC-DAD method was further confirmed by HPLC-ESI-MS/MS system in multiple reaction monitoring mode. Each separated HPLC peak was identified as the target alkaloid, showing its relevant ionized molecule and selected fragment ion. By applying the established method, alkaloid production during the E. californica cell culture could be successfully monitored and some valuable information on its metabolism could be deduced.

  14. QuEChERS-HPLC-DAD method for sulphonamides in chicken breast

    Directory of Open Access Journals (Sweden)

    Simone Caetani Machado

    2013-03-01

    Full Text Available The development of a QuEChERS-HPLC-DAD method using a Lichrospher 60 RP-Select B column (250 x 4.6 mm x 5 µm at 40ºC, mobile phase constituted by phosphate buffer:acetonitrile (75:25, v/v at a initial flow rate of 0.5 mL min-1, increased by 1.2 mL min-1 and at 265 nm is presented for simultaneous determination of sulphadiazine, sulphametoxipiridazine and sulphamethoxazole in chicken breast samples. QuEchERS is inexpensive, fast and easy, and the extraction of the analytes of the matrix was successfully employed. In addition, the method presented linearity, in the range of 25, 50, 100, 150, 175, and 200 µg kg-1, precision, selectivity and sensitivity. The intraday precision (RSD % for QuEChERS method was between 3.6-10.8 (SDZ, 6.9-14.1 (SPZ and 1.9-10.9 (SMX and interday precision (RSD% was between 1.5-9.7, 1.7-4.1 and 2.1-10.2, respectively. Results of accuracy (bias were in the range of -8.6 to +11.9 %. Therefore, the validated method is clearly useful for the practical residue monitoring of the drugs evaluated in chicken samples, as all the values were within the acceptable criteria used for food safety. Of 6 samples analyzed, none of them showed contamination of the sulphonamides studied at detectable levels.O desenvolvimento de um método QuEChERS-HPLC-DAD usando uma coluna Lichrospher RP-60 Select B (250 x 4,6 mm x 5 µm a 40 ºC, fase móvel constituída por tampão de fosfato: acetonitrila (75:25, v/v a uma vazão inicial de 0,5 mL min-1, aumentando 1,2 mL min-1 e a 265 nm é apresentado para a determinação simultânea de sulfadiazina, sulfametoxipiridazina e sulfametoxazol em amostras de peito de frango. O QuEChERS é barato, rápido e fácil, e a extração dos analitos da matriz foi empregada com sucesso. Além disso, o método apresentou linearidade, na faixa de 25, 50, 100, 150, 175 e 200 µg kg-1, precisão, seletividade e sensibilidade. A precisão intradia (RSD % para o método QuEChERS foi entre 3,6-10,8 (SDZ, 6,9-14,1 (SPZ

  15. 不同品种石斛的HPLC-DAD-ELSD指纹图谱研究%Study on HPLC-DAD-ELSD Fingerprint in Different Species of Dendrobii Caulis

    Institute of Scientific and Technical Information of China (English)

    石媛慧; 郭力; 许莉; 郝普彦; 陈佳江; 韦练; 罗方利; 张廷模; 韩瑜

    2013-01-01

    目的:采用HPLC-DAD-ELSD建立石斛指纹图谱,并对金钗、鼓槌、叠鞘、细茎和反瓣等5种石斛的指纹图谱进行对比研究。方法:采用Dikma C18(250 mm ×4.6 mm,5µm)色谱柱,流动相为乙腈-0.5%冰乙酸水溶液,梯度洗脱,柱温30℃,流速1 mL·min-1,DAD串联蒸发光散射检测器检测。结果:建立5种石斛的HPLC指纹图谱,并标定9个指纹峰,5种石斛相似度较低。结论:建立的指纹图谱方法稳定、可靠、重现性好,可直接反映石斛品种与化学成分变化的关系。%This study was aimed to set up the fingerprint chromatograms of chemical principles of Dendrobii Caulis by HPLC-DAD-ELSD. This analysis was performed at 30°C on a Dikma Cl8 column (250 mm × 4.6 mm, 5 µm) with a mobile phase consisting of acetonitrile-0.5% glacial acetic acid, gradient elution, at the flow rate of 1.0 mL•min-1 with DAD and ELSD detection. The HPLC fingerprint of Dendrobii Caulis was established. And 9 fingerprint peaks were marked. The similarity among different species was low. This method was stable, reliable and repeatable. It is useful for the study on the relationship between chemical components and species of Dendrobii Cauli.

  16. Investigation of the Key Pharmacological Activities of Ficus racemosa and Analysis of Its Major Bioactive Polyphenols by HPLC-DAD

    Science.gov (United States)

    Sumi, Salma Akter; Siraj, Md. Afjalus; Hossain, Amir; Mia, Md. Sagir; Afrin, Seagufta

    2016-01-01

    Objective. Oxidative stress leads to numerous physiological disorders including infectious diseases, inflammation, and cancer. The present study was carried out to investigate antioxidant, antibacterial, and cytotoxic activity of methanol crude extract of leaves and fruits of the Ficus racemosa (LCME and FCME, resp.) and to analyse its major bioactive polyphenols by HPLC-DAD. Methods. Antioxidant capacity of the extracts was evaluated by DPPH free radical scavenging, reducing power, total phenolic, total flavonoid, total tannin content assay, superoxide radical, hydroxyl radical, and hydrogen peroxide scavenging assay. Identification and quantification of bioactive polyphenols were done by HPLC-DAD method. Antibacterial activity was tested by “disc diffusion” method. Brine shrimp lethality assay was carried out to check the cytotoxic potential. Result. Both LCME and FCME showed DPPH scavenging ability and concentration dependent reducing power activity. They had phenolic content, flavonoid content, and tannin content. Both the extracts showed superoxide radical scavenging ability, hydroxyl radical scavenging ability, and hydrogen peroxide scavenging ability. HPLC analysis of LCME and FCME indicated the presence of significant amount of gallic acid along with other phenolic constituents. Conclusion. Significant amount of gallic acid along with other phenolic constituents might have played an important role in the observed antioxidant, antibacterial, and cytotoxic activity. PMID:28105059

  17. [Study on limit detection of flavones in diterpene ginkgolides meglumine injection materials by LC-MS and HPLC-DAD].

    Science.gov (United States)

    Bi, Sen; Li, Yan-jing; Huang, Wen-zhe; Kang, Dan-yu; Ding, Gang; Xiao, Wei

    2015-08-01

    Limit test of flavones in diterpene ginkgolides meglumine injection materials by UV-Vis and HPLC-DAD method was studied in this essay. The HPLC-DAD method has lower LOD (about 1% of the UV-Vis), that is, the sensitivity is higher than UV-Vis method. Through the analysis of the kinds of flavonoids ingredients in the samples by LC-MS, the three compounds with highest contents are kaempferol, quercetin and isorhamnetin. Kaempferol, quercetin and isorhamnetin were chosen as reference compounds for HPLC analysis, and the HPLC separation analysis was carried on an Agilent Eclipse plus C18 column (4.6 mm x 250 mm, 5 μm) with methanol and water containing 0.4% phosphoric acid (50: 50) as mobile phase, and the flow rate was 1.0 mL x min(-1). The detection wavelength was set at 360 nm. This method has good specificity, precision and reproducibility. The LODs of quercetin, kaempferide and isorhamnetin were 27.6, 22.3, 29.5 μg x L(-1). The average recovery was 87.9% (RSD 3.3%), 91.7% (RSD 3.1%), 88.3 (RSD 1.3%) for quercetin, kaempferide and isorhamnetin, respectively. Based on the 10 batches of sample results and sensitivity of different HPLC, the content of total flavonoids ingredients of diterpene ginkgolides meglumine injection materials was limited no more than 2 x 10(-5). This method is simple, quick and has good maneuverability, and could be used to the limit test of flavonoids in the diterpene ginkgolides meglumine injection materials.

  18. Simultaneous qualitative and quantitative analysis of phenolic acids and flavonoids for the quality control of Apocynum venetum L. leaves by HPLC-DAD-ESI-IT-TOF-MS and HPLC-DAD.

    Science.gov (United States)

    An, Haijuan; Wang, Hong; Lan, Yuexiang; Hashi, Yuki; Chen, Shizhong

    2013-11-01

    A reliable method based on high performance liquid chromatography-diode array detector-electrospray ionization-ion trap-time of flight-mass spectrometry (HPLC-DAD-ESI-IT-TOF-MS) was developed for the identification of phenolic acids and flavonoids in Apocynum venetum L. leaves and its adulterant, Pocynum hendersonii (Hook. f.) Woodson leaves. A total of 21 compounds were identified or tentatively identified, including 4 phenolic acids and 17 flavonoids. 3-O-caffeoylquinic acid (3-CQA) and caffeic acid were detected for the first time in A. venetum leaves; 4-O-caffeoylquinic acid (4-CQA), 3-CQA, caffeic acid, quercetin-3-O-(6"-O-malonyl)-galactoside, quercetin-3-O-(6"-O-malonyl)-glucoside, kaempferol-3-O-(6"-O-malonyl)-glucoside, kaempferol-3-O-(6"-O-acetyl)-glucoside, and kaempferol-3-O-dihexoside were detected for the first time in P. hendersonii leaves. Cluster analysis was employed to analyze 24 batches of A. venetum leaves and 5 batches of P. hendersonii leaves collected from various regions in China. The analysis, which was based on the 21 compounds, indicated that profiles of these compounds were distinct between the two species, and among A. venetum leaf samples from different origins. 18 of these 21 compounds were selected as the markers and simultaneously analyzed by HPLC-DAD for the first time. The quantitative analytical method was validated and subsequently applied to the comprehensive quality evaluation of 24 batches of A. venetum leaves.

  19. Qualitative and quantitative measurement of cannabinoids in cannabis using modified HPLC/DAD method.

    Science.gov (United States)

    Patel, Bhupendra; Wene, Daniel; Fan, Zhihua Tina

    2017-08-10

    This study presents an accurate and high throughput method for the quantitative determination of various cannabinoids in cannabis plant material using high pressure liquid chromatography (HPLC) with a diode array detector (DAD). Sample extraction and chromatographic analysis conditions for the measurement of cannabinoids in the complex cannabis plant material matrix were optimized. The Agilent Poroshell 120 SB-C18 column provided high resolution for all target analytes with a short run time (10minutes) given the core shell technology. The aqueous buffer mobile phase was optimized with ammonium acetate at pH 4.75. The change in the mobile phase and the new column ensured a separation between cannabidiol (CBD and cannabigerol (CBG) along with cannabigerol and tetrahydrocannabinolic acid (THCA), which were not well separated by previous publications, improved buffering capacity, and provided analytical performance stability. Moreover, baseline drifting was significantly minimized by the use of a low concentration buffer solution (25mM ammonium acetate). In addition, evaporation and reconstitution of the sample residue with a methanol-organic pure (OP) water solution (65:35) significantly reduced the matrix interference. The modified extraction produced good recoveries (>91%) for each of the eight cannabinoids. The optimized method was validated for specificity, linearity, sensitivity, precision, accuracy, and stability. The combined relative standard deviation (%RSD) for intra-day and inter-day precision for all eight analytes varied from 2.5% to 5.2% and 0.28% to 5.5%, respectively. The %RSD for the repeatability study varied from 1.1% to 5.5%. The recoveries from spiked cannabis matrix samples were greater than 90% for all analytes, except delta-8-tetrahydrocannabinol (Δ(8)-THC), which was 80%. The recoveries varied from 81% to 107% with a precision of 0.7-8.1%RSD. Delta-9-tetrahydrocannabinol (Δ(9)-THC) in all of the cannabis samples (n=635) was less than 10

  20. HPLC-DAD Phenolic Characterization and Antioxidant Activities of Ripe and Unripe Sweet Orange Peels

    Science.gov (United States)

    Omoba, Olufunmilayo Sade; Obafaye, Rebeccah Olajumoke; Salawu, Sule Ola; Boligon, Aline Augusti; Athayde, Margareth Linde

    2015-01-01

    Phenolic compounds of unripe and ripe sweet orange peels were determined using a high-performance liquid chromatography separation method with diode array detector (HPLC-DAD). The in vitro antioxidant properties and the EC50 (concentration required to obtain a 50% antioxidant effect) values were also determined. The predominant phenolic compounds were quercitrin, rutin, and quercetin with values of 18.77 ± 0.01 mg/mL, 18.65 ± 0.03 mg/mL, and 10.39 ± 0.01 mg/mL respectively in unripe orange peel and 22.61 ± 0.01 mg/mL, 17.93 ± 0.03 mg/mL, and 14.03 ± 0.02 mg/mL respectively in ripe orange peel. The antioxidant properties revealed 2,2′-azino-bis(3-ethyl benzothiazoline-6-sulfonic acid) diammonium salt (ABTS) scavenging ability of both unripe and ripe orange peels respectively as 14.68 ± 0.01 and 16.89 ± 0.02 mmol TEAC/g, the Ferric Reducing Antioxidant Properties (FRAP) as 70.69 ± 0.01 and 91.38 ± 0.01 mg gallic acid equivalents/100g, total phenol content as 5.27 ± 0.03 and 9.40 ± 0.01 mg gallic acid equivalents/g and total flavonoid content as 3.30 ± 0.30 and 4.20 ± 0.02 mg quercetin equivalent/g. The antioxidant assays showed enhanced potency of extract from ripe orange peel with EC50 values of 2.71 ± 0.03 mg/mL for 2,2-diphenyl-1-picrylhydrazyl (DPPH), 0.67 ± 0.03 mg/mL for hydroxyl radicals (OH*), 0.57 ± 0.02 mg/mL for Fe2+ chelation, and 0.63 ± 0.06 mg/mL for malondialdehyde (MDA), and was more potent than unripe orange peel. PMID:26783839

  1. Quantitative analysis combined with chromatographic fingerprint for comprehensive evaluation of Xiaoer Chaigui Tuire granules by HPLC-DAD.

    Science.gov (United States)

    Liu, Hong-Ming; Nie, Lei

    2015-01-01

    Quantitative analysis of eight major components combined with chromatographic fingerprint based on high performance liquid chromatography coupled with diode array detector (HPLC-DAD) was developed for the quality evaluation of Xiaoer Chaigui Tuire granules (XCTG), a traditional Chinese medicine (TCM) preparation. Each compound was analyzed by comparing its retention time and UV spectrum of each chromatographic peak with the corresponding retention time and UV spectrum of each standard compound. Baseline separation was achieved on an Agilent Zorbax SB-C18 column with gradient elution of acetonitrile and 0.1% (v/v) phosphoric acid. The developed method was validated by linearity, precision, repeatability, stability and recovery and was subsequently applied to quality evaluation of 12 batches of XCTG with similarity analysis, principal component analysis and cluster analysis. Quantitative analysis combined with HPLC fingerprint could offer an efficient, reliable and practical approach for quality evaluation of XCTG.

  2. HPLC-DAD-ESI-MS analysis of phenolic compounds during ripening in exocarp and mesocarp of tomato fruit.

    Science.gov (United States)

    Carrillo-López, Armando; Yahia, Elhadi

    2013-12-01

    Identification of phenolic compounds was done by means of liquid chromatography (HPLC) coupled to mass spectrometry (MS) using the electrospray ionization interface (ESI). Quantification of phenolic compounds was carried out by using HPLC with diode array detector (DAD) in exocarp and mesocarp of tomato fruit at 6 different ripeness stages (mature-green, breakers, turning, pink, light-red, and red). Several phenolic compounds were identified including chlorogenic acid, caffeic acid, p-coumaric acid, ferulic acid, and rutin and some combined phenolic acids were tentatively identified, mainly glycosides, such as caffeoyl hexose I, caffeoyl hexose II, caffeoylquinic acid isomer, dicaffeoylquinic acid, p-coumaroyl hexose I, p-coumaroyl hexose II, feruloyl hexose I, feruloyl hexose II, siringyl hexose, and caffeoyl deoxyhexose hexose. Fruit exocarp had higher quantities of total soluble phenolics (TSP) compared to mesocarp. During ripening, TSP increased in both exocarp and mesocarp, mainly in exocarp. While rutin increased, chlorogenic acid decreased in both tissues: exocarp and mesocarp.

  3. HPLC-DAD-ESI/MS(n) analysis of phenolic compounds for quality control of Grindelia robusta Nutt. and bioactivities.

    Science.gov (United States)

    Ferreres, Federico; Grosso, Clara; Gil-Izquierdo, Angel; Valentão, Patrícia; Azevedo, Carolina; Andrade, Paula B

    2014-06-01

    The phenolic composition of herbal tea (HT) and hydromethanolic extract (HME) obtained from Grindelia robusta Nutt. was studied by HPLC-DAD-ESI/MS(n). Thirty six flavonoids and hydroxycinnamic acids were detected, from which thirty are described for the first time in this species. Quantification by HPLC-DAD showed that diosmetin-7-O-glucuronide-3'-O-pentoside+apigenin-7-O-glucuronide-4'-O-pentoside, apigenin-7-O-glucuronide+diosmetin-7-O-glucuronide and 3,5-dicaffeoylquinic acid+3,4-dicaffeoylquinic acid were the major compounds. Since the health-promoting effects of natural phenolic compounds against brain disorders is of increasing interest, HT and HME were also tested against oxygen and nitrogen reactive species and against enzymes related with Alzheimer's disease and depression. Extracts displayed strong in vitro scavenging activity and monoamine oxidase-A (MAO-A) inhibitory activity. The anti-MAO-A capacity was observed at non-toxic concentrations for SH-SY5Y human neuroblastoma cell line, reinforcing the benefits of G. robusta HT. However, no protection against hydrogen peroxide treatment was observed.

  4. A Simple HPLC-DAD Method for the Analysis of Melamine in Protein Supplements: Validation Using the Accuracy Profiles

    Directory of Open Access Journals (Sweden)

    Domenico Montesano

    2013-01-01

    Full Text Available The study presents a fully validated simple high-performance liquid chromatography method with diode array detection (HPLC-DAD, able to accurately determine the melamine, fraudulently added, in protein supplements, commonly used from healthy adults to enhance exercise or sport performance. The validation strategy was intentionally oriented towards routine use and the reliability of the method rather than extreme performance. For this reason, validation by accuracy profile, including estimation of uncertainty, was chosen. This procedure, based on the concept of total error (bias + standard deviation, clearly showed that this method was able to determine melamine over the range of 0.05–3.0 mg Kg−1, selected by taking into account the maximum residue levels (MRLs proposed by European legislation to distinguish between the unavoidable background presence of melamine and unacceptable adulteration. The accuracy profile procedure established that at least 95% of the future results obtained with the proposed method would be within the ±15% acceptance limits of the validated HPLC-DAD method over the whole defined concentration range.

  5. Rapid Identification and Assignation of the Active Ingredients in Fufang Banbianlian Injection Using HPLC-DAD-ESI-IT-TOF-MS.

    Science.gov (United States)

    Li, Sensen; Lin, Zongtao; Jiang, Haixiu; Tong, Lingkun; Wang, Hong; Chen, Shizhong

    2016-08-01

    Fufang Banbianlian Injection (FBI) is a well-known traditional Chinese medicine formula composed of three herbal medicines. However, the systematic investigation on its chemical components has not been reported yet. In this study, a high-performance liquid chromatography combined with diode-array detector, and coupled to an electrospray ionization with ion-trap time-of-flight mass spectrometry (HPLC-DAD-ESI-IT-TOF-MS) method, was established for the identification of chemical profile in FBI. Sixty-six major constituents (14 phenolic acids, 14 iridoids, 20 flavonoids, 2 benzylideneacetone compounds, 3 phenylethanoid glycosides, 1 coumarin, 1 lignan, 3 nucleosides, 1 amino acids, 1 monosaccharides, 2 oligosaccharides, 3 alduronic acids and citric acid) were identified or tentatively characterized by comparing their retention times and MS spectra with those of standards or literature data. Finally, all constituents were further assigned in the individual herbs (InHs), although some of them were from multiple InHs. As a result, 11 compounds were from Lobelia chinensis Lour, 33 compounds were from Scutellaria barbata D. Don and 38 compounds were from Hedyotis diffusa Willd. In conclusion, the developed HPLC-DAD-ESI-IT-TOF-MS method is a rapid and efficient technique for analysis of FBI sample, and could be a valuable method for the further study on the quality control of the FBI.

  6. Comparison of piracetam measured with HPLC-DAD, HPLC-ESI-MS, DIP-APCI-MS, and a newly developed and optimized DIP-ESI-MS.

    Science.gov (United States)

    Lenzen, Claudia; Winterfeld, Gottfried A; Schmitz, Oliver J

    2016-06-01

    The direct inlet probe-electrospray ionization (DIP-ESI) presented here was based on the direct inlet probe-atmospheric pressure chemical ionization (DIP-APCI) developed by our group. It was coupled to an ion trap mass spectrometer (MS) for the detection of more polar compounds such as degradation products from pharmaceuticals. First, the position of the ESI tip, the gas and solvent flow rates, as well as the gas temperature were optimized with the help of the statistic program Minitab® 17 and a caffeine standard. The ability to perform quantitative analyses was also tested by using different concentrations of caffeine and camphor. Calibration curves with a quadratic calibration regression of R (2) = 0.9997 and 0.9998 for caffeine and camphor, respectively, were obtained. The limit of detection of 2.5 and 1.7 ng per injection for caffeine and camphor were determined, respectively. Furthermore, a solution of piracetam was used to compare established analytical methods for this drug and its impurities such as HPLC-diode array detector (DAD) and HPLC-ESI-MS with the DIP-APCI and the developed DIP-ESI. With HPLC-DAD and 10 μg piracetam on column, no impurity could be detected. With HPLC-ESI-MS, two impurities (A and B) were identified with only 4.6 μg piracetam on column, while with DIP-ESI, an amount of 1.6 μg piracetam was sufficient. In the case of the DIP-ESI measurements, all detected impurities could be identified by MS/MS studies. Graphical Abstract Scheme of the DIP-ESI principle.

  7. Analysis of black carbon molecular markers by two chromatographic methods (GC-FID and HPLC-DAD)

    Science.gov (United States)

    Schneider, Maximilian P. W.; Smittenberg, Rienk H.; Dittmar, Thorsten; Schmidt, Michael W. I.

    2010-05-01

    The analysis of benzenepolycarboxylic acids (BPCA) as a quantitative measure for black carbon (BC) in soil and sediment samples is a well-established method [1, 2]. Briefly, the oxidation of polycondensated BC molecules forms seven molecular markers, which can be assigned to BC, and which subsequently can be quantified by GC-FID (gas chromatography with flame ionization detector). Recently this method has been refined for BC quantification in seawater samples measuring BPCA on HPLC-DAD (High performance liquid chromatography with diode array detector) [3]. However, a systematic comparison of BC as determined by both analytical techniques would be essential to the calculation of global BC budgets, but is lacking. Here we present data for the systematic comparison of the two BPCA methods, both for quantity and quality. We prepared chars under well-defined laboratory conditions. Chestnut hardwood chips and rice straw were pyrolysed at temperatures between 200 and 1000°C under constant N2 stream. The BC contents of the chars have been analysed using the BPCA extraction method followed by either GC-FID or HPLC-DAD quantification [4]. It appears that the GC-FID method yields systematically lower concentrations of BPCA in the chars compared to the HPLC-DAD method. Possible reasons for the observed difference are i) higher losses of sample material during preparation for GC-FID; ii) different quality of the linear regression used for quantification; iii) incomplete derivatisation of B5CA and B6CA, which is needed for GC-FID analysis. In a next step, we will test different derivatisation procedures (methylation with dimethyl sulfate or diazomethane, and silylation) for their influence on the GC-FID results. The aim of this study is to test if black carbon can be quantified in soil, sediment and water samples using one single method - a crucial step when attempting a global BC budget. References: [1] Brodowski, S., Rodionov, A., Haumeier L., Glaser, B., Amelung, W. (2005

  8. Simultaneous determination of eight phenolic acids, five saponins and four tanshinones for quality control of compound preparations containing Danshen-Sanqi herb-pair by HPLC-DAD

    Directory of Open Access Journals (Sweden)

    Hong Yao

    2017-01-01

    Abbreviations used: DS: Salviae miltiorrhizae; SQ: Panaxnotoginseng; HPLC: high-performance liquid chromatography; DAD: diode array detector; LOD: limit of detection; LOQ: limit of quantification; TCMs: Traditional Chinese medicines; GDDP: Guanxin Danshen dripping pills; FDDP: Fufang Danshen dripping pills; FDT: Fufang Danshen tablets; FDC: Fufang Danshen capsules; GP: Guanxin pills

  9. HPLC-DAD-MS鉴定注射用雷贝拉唑钠杂质及其特征谱研究%Study on the impurities in rabeprazole sodium for injection using HPLC-DAD-MS

    Institute of Scientific and Technical Information of China (English)

    孙国祥; 丁楠; 王真; 宋良伟

    2011-01-01

    目的 利用高效液相色谱-二极管阵列检测器-质谱联用方法对注射用雷贝拉唑钠中杂质成分进行分析和结构推断.方法 采用CenturySILc18 BDS色谱柱(250 mm×4.6 mm,5 μm),以磷酸盐缓冲液(磷酸氢二钠1.119 g和磷酸二氢钠0.179 g水溶液)-乙腈(体积比60:40)为流动相,检测波长286 nm,流速1.0 mL·min-1,柱温(35±0.15)℃,进样量10μL,检测雷贝拉唑钠及其杂质;通过DAD检测器和质谱,获得在线紫外光谱图和质谱数据.结果 通过在线紫外光谱图和质谱结果推断出可能含有的杂质结构,并测定了主要杂质含量限度.结论 高效液相色谱-二极管阵列检测器-质谱联用技术可用于注射用雷贝拉唑钠中杂质的定性推断和定量测定.%Objective To analyze and infer the impurities' structures in rabeprazole sodium for injection by HPLC-DAD-MS. Methods The operation was carried out on a CenturySIL C18 BDS column (250 mm × 4. 6 mm,5 μm), with the mobile phase consisting of a mixture of phosphate buffer ( disodium hydrogen phosphate 1. 119 g and sodium dihydrogen phosphate 0. 179 g)-acetonitrile (V: V = 60: 40). The detection wavelength was set at 286 nm, the flow rate was 1.0 mL· min-1, the column temperature was ( 35 ± 0. 15)℃ and the injectionvolume was 10 μL,the rabeprazole sodium and its impurities were detected,the online UV spectrum and MS data were obtained by using DAD detector and MS. Results According to the results of online UV spectra and mass spectrometry ,the feasible impurities' structures were inferred and the limits of the main impurities were determined. Conclusions The method of HPLC-DAD-MS can be used to identify the impurities in rabeprazole sodium for injection.

  10. Assessing Jasminum grandiflorum L. authenticity by HPLC-DAD-ESI/MS(n) and effects on physiological enzymes and oxidative species.

    Science.gov (United States)

    Ferreres, Federico; Grosso, Clara; Gil-Izquierdo, Angel; Valentão, Patrícia; Andrade, Paula B

    2014-01-01

    The dried flower buds of Jasminum grandiflorum L. are widely consumed as infusion and used in traditional medicine for psychiatric disorders. It is important to have a well-established method for the chemical characterization of J. grandiflorum since there are resemblances with a toxic species, Gelsemium sempervirens (L.) Jaume Saint-Hilaire. HPLC-DAD-ESI/MS(n) analysis allowed identifying six phenolic compounds for the first time. Moreover, the evaluation of in vitro activity against central nervous system (CNS) related enzymes was undertaken for the first time, as well as against reactive species in order to support the efficacy towards CNS disorders. Copyright © 2013 Elsevier B.V. All rights reserved.

  11. Simultaneous determination of nine components in Qingkailing injection by HPLC/ELSD/DAD and its application to the quality control.

    Science.gov (United States)

    Yan, Shikai; Luo, Guoan; Wang, Yiming; Cheng, Yiyu

    2006-03-01

    High-performance liquid chromatography coupled with photo diode array detection and evaporative light scattering detection (HPLC/DAD/ELSD) was established to simultaneously determine nine ingredients in Qingkailing injection. Four wavelengths at 240, 254, 280 and 330 nm, respectively, were chosen as the monitoring wavelength to determine two nucleosides (uridine and adenosine), geniposide, baicalin and two organic acids (chlorogenic acid and caffeic acid), and an evaporative light scattering detector combined was employed to determine three steroids (cholic acid, ursodeoxycholic acid and hyodeoxycholic acid). This assay was fully validated in respect to precision, repeatability and accuracy. The proposed method was successfully applied to quantify the nine ingredients in 19 different Qingkailing injection samples and by principal component analysis (PCA) and hierarchical clustering analysis (HCA), it demonstrated significant variations in the content of these compounds in the samples from different manufacturers and preparation procedures. This method could be readily utilized as a quality control method for traditional Chinese medicine (TCM).

  12. Determination of free and esterified carotenoid composition in rose hip fruit by HPLC-DAD-APCI(+)-MS.

    Science.gov (United States)

    Zhong, Lijie; Gustavsson, Karl-Erik; Oredsson, Stina; Głąb, Bartosz; Yilmaz, Jenny Lindberg; Olsson, Marie E

    2016-11-01

    Rose hip fruit, which contains high concentration of carotenoids is commonly used for different food products in Europe and it is considered to have medical properties. In this study, a simple, rapid and efficient HPLC-DAD-APCI(+)-MS method was developed and applied to identify and quantify the carotenoids in rose hip fruit of four rose species, including both unsaponified and saponified extract. In the unsaponified extract 23 carotenoid esters were detected, in which either rubixanthin ester or violaxanthin ester was the dominant component of the ester composition. In the saponified extract 21 carotenoids, including 11 xanthophylls and 10 carotenes were detected. This is the first time the total carotenoid composition, including the carotenoid esters in rose hip fruit were identified and quantified. This work reveals the potential of rose hip fruit to be utilized as a healthy dietary material and give chemical information for the possible future development in the pharmacology field.

  13. Effect of the addition of flavan-3-ols on the HPLC-DAD salivary-protein profile.

    Science.gov (United States)

    Quijada-Morín, Natalia; Crespo-Expósito, Carlos; Rivas-Gonzalo, Julián C; García-Estévez, Ignacio; Escribano-Bailón, María Teresa

    2016-09-15

    The interaction between monomeric flavan-3-ols and salivary proteins has been studied using HPLC-DAD. A chromatographic method has been described and seven protein fractions were collected. The peptides and proteins present in each fraction have been identified using nLC-MS-MS analysis. The interaction between saliva and catechin, epicatechin and gallocatechin has been studied. These compounds interact in a discriminated way with salivary proteins: catechin causes a decrease of some fractions, epicatechin causes the decrease or increase of fractions while gallocatechin seems to cause an increase of two fractions. This variable behavior is explained, for the decrease in the chromatographic area, by the precipitation of salivary proteins and, for the increase of the area, by the formation of soluble complexes and/or for the formation of new peaks.

  14. GC-FID and HPLC-DAD Methods for the Determination of Menadione Sodium Bisulphite Directly and by Converting Menadione Sodium Bisulphite to Menadione in Pharmaceutical Preparation.

    Science.gov (United States)

    Demirkaya-Miloglu, Fatma; Kadioglu, Yucel; Senol, Onur

    2014-01-01

    was performed in both direct analysis of MSB and analysis of MN by converting MSB to MN with sodium carbonate. GC-FID method was carried out on the HP-5 capillary column GC-FID and HPLC-DAD methods were developed for determination of menadione (MN) and menadione sodium bisulphite (MSB). By means of each method, quantitative analysis of MSB in commercial pharmaceutical using nitrogen gas. HPLC-DAD method was achieved on the reversed phase C8 column by using a mobile phase consisting methanol and water. The calibration curves of GC-FID and HPLC-DAD for both analytes were linear in the same concentration range (0.5-20 μg/mL). Both methods were validated in terms of precision, accuracy, recovery and limits of detection (LOD) and quantitation (LOQ). Although LOD values of HPLC-DAD method (0.010 μg/mL for MN and 0.005 μg/mL for MSB) is lower than obtained values with GC-FID method (0.04 μg/mL for MN and 0.06 μg/mL for MSB), both methods gave similar and favorable results in terms of precision and accuracy. The Student's t-test was applied to investigate the significant of the different between the results of MSB determination with direct analysis of MSB and analysis of MN by converting MSB to MN by means of GC-FID and HPLC-DAD method in dosage form.

  15. HPLC-DAD and UV-spectrophotometry for the determination of lychnopholide in nanocapsule dosage form: validation and application to release kinetic study.

    Science.gov (United States)

    Branquinho, Renata Tupinambá; Mosqueira, Vanessa Carla Furtado; Kano, Eunice Kazue; de Souza, Jacqueline; Dorim, Diego Dias Ramos; Saúde-Guimarães, Dênia Antunes; de Lana, Marta

    2014-01-01

    Simple and sensitive methods using high-performance liquid chromatography-diode array detection (HPLC-DAD) and ultraviolet (UV)-spectrophotometry were developed and compared to quantify lychnopholide (LYC) in poly-ε-caprolactone nanocapsules and to study its release kinetics. Both methods were validated concerning their specificity, linearity, limits of detection and quantification, precision, accuracy and stability. HPLC-DAD analyses were conducted using an RP C18 column, isocratic elution with a methanol-water (60:40 v/v) mobile phase at 0.8 mL/min flow rate and detection at 265 nm. The linear response (r(2) > 0.999) was obtained within a concentration range of 2-25 µg/mL using HPLC-DAD and 5-40 µg/mL using spectrophotometry. Intra-day and inter-day precision were obtained with low relative standard deviation values. The accuracy of the methods was within the range 98-101% for HPLC-DAD and from 96-100% for UV-spectrophotometry. Both methods were suitable to be applied for the determination of drug loading percentage (>96%) and encapsulation efficiency (>90%). Furthermore, the sensitivity of HPLC-DAD method allows studies of LYC release/dissolution in sink conditions. LYC presented 100% dissolution after 24 h, whereas only 60% of LYC was released from the nanocapsule dosage form, with no burst effect. The methods fulfilled all validation parameters evaluated for LYC quantification in the polymeric nanocapsules and have proven to be accurate, selective and sensitive in the previously mentioned applications.

  16. Phytochemical analysis of Rosa hybrida cv. 'Jardin de Granville' by HPTLC, HPLC-DAD and HPLC-ESI-HRMS: polyphenolic fingerprints of six plant organs.

    Science.gov (United States)

    Riffault, Ludivine; Destandau, Emilie; Pasquier, Laure; André, Patrice; Elfakir, Claire

    2014-03-01

    The Rosa hybrida cultivar 'Jardin de Granville', a delicate clear pink flower, is here investigated through a progressive analytical strategy using complementary phytochemical screening methods in order to characterize the polyphenol content of several parts of the plant. The microwave hydro-ethanolic extract analysis of six different parts of the plant, carried out by High Performance Thin Layer Chromatography (HPTLC) and High Performance Liquid Chromatography coupled with a Diode Array Detector (HPLC-DAD) enabled initial identification of the polar molecular families present in each organ, namely tannins and flavonoids (quercetin and kaempferol derivatives). The HPLC fingerprints displayed different profiles for each organ, attesting to the original composition and potential valuation of the different plant parts. More detailed analyses of the extracts were then carried out by High Performance Liquid Chromatography coupled with electrospray ionization (ESI) mass spectrometry with a Q-TOF analyzer (ESI-HR-Q-TOF). Around 60 compounds were identified, mainly gallo-tannins, ellagi-tannins, catechin derivatives and glycoside derivatives of quercetin and kaempferol. Some compounds such as hyperoside or ellagic acid appeared to be ubiquitous and were found in abundance in each plant part. Woods were the richest organ in catechin and proanthocyanidin derivatives while kaempferol derivatives were more numerous and abundant in bud and flower parts. Copyright © 2014 Elsevier Ltd. All rights reserved.

  17. Identification and Analysis of Amygdalin, Neoamygdalin and Amygdalin Amide in Different Processed Bitter Almonds by HPLC-ESI-MS/MS and HPLC-DAD.

    Science.gov (United States)

    Xu, Shuya; Xu, Xinfang; Yuan, Shaoxiong; Liu, Huan; Liu, Mengnan; Zhang, Ying; Zhang, Hui; Gao, Yan; Lin, Ruichao; Li, Xiangri

    2017-08-30

    Processing is a traditional pharmacy technology based on traditional Chinese medicine theory. The traditional Chinese medicine (TCM) ingredients should be processed before being used as a medicine. Processed bitter almonds are widely used in the clinic in TCM for the treatment of cough and asthma. In this work the amygdalin profile of three producing areas in China was determined, with respect to three differently processed bitter almond products: raw, stir-fried and scalded. Identification of the compounds was done by using high performance liquid chromatography coupled to electrospray ionization mass spectrometry (HPLC-ESI-MS/MS). Results indicated that amygdalin, neoamygdalin and amygdalin amide were identified in the different processed bitter almonds. Meanwhile, amygdalin was used as a standard to calculate the quantification of amygdalin and the concentration ratio of neoamygdalin and total amygdalin by HPLC-DAD. The data suggested that composition of amygdalin isomers in bitter almonds was influenced by the processing method. It also gives a new understanding of the processing principle of bitter almonds. Moreover, the classification of different processed bitter almonds can be achieved on the basis of amygdalin isomers levels.

  18. Characterisation of Phenolic Compounds in South African Plum Fruits (Prunus salicina Lindl. using HPLC Coupled with Diode-Array, Fluorescence, Mass Spectrometry and On-Line Antioxidant Detection

    Directory of Open Access Journals (Sweden)

    Dalene de Beer

    2013-05-01

    Full Text Available Phenolic compounds are abundant secondary metabolites in plums, with potential health benefits believed to be due to their antioxidant activity, amongst others. Phenolic characterisation of South African Prunus salicina Lindl. plums is necessary to fully evaluate their potential health benefits. An HPLC method using diode-array detection (DAD for quantification of phenolic compounds was improved and fluorescence detection (FLD was added for quantification of flavan-3-ols. Validation of the HPLC-DAD-FLD method showed its suitability for quantification of 18 phenolic compounds, including flavan-3-ols using FLD, and phenolic acids, anthocyanins and flavonols using DAD. The method was suitable for characterisation of the phenolic composition of 11 South African plum cultivars and selections, including various types with yellow and red skin and flesh. The method was used in conjunction with mass spectrometry (MS to identify 24 phenolic compounds. Neochlorogenic acid and cyanidin-3-O-glucoside were the major compounds in most of the plums, while cyanidin-3-O-glucoside was absent in Sun Breeze plums with yellow skin and flesh. Post-column on-line coupling of the ABTS•+ scavenging assay with HPLC-DAD enabled qualitative evaluation of the relative contribution of individual phenolic compounds to the antioxidant activity. The flavan-3-ols, neochlorogenic acid and cyanidin-3-O-glucoside displayed the largest antioxidant response peaks.

  19. A Comparative Study of Newly Developed HPLC-DAD and UHPLC-UV Assays for the Determination of Posaconazole in Bulk Powder and Suspension Dosage Form

    Directory of Open Access Journals (Sweden)

    Dalia A. Hamdy

    2014-01-01

    Full Text Available Objective. To develop and compare HPLC-DAD and UHPLC-UV assays for the quantitation of posaconazole in bulk powder and suspension dosage form. Methods. Posaconazole linearity range was 5–50 μg/mL for both assays. For HPLC-DAD assay, samples were injected through Zorbax SB-C18 (4.6 × 250 mm, 5 μm column. The gradient elution composed of the mobile phase acetonitrile: 15 mM potassium dihydrogen orthophosphate (30 : 70 to 80 : 20, linear over 7 minutes pumped at 1.5 mL/min. For UHPLC-UV assay, samples were injected through Kinetex-C18 (2.1 × 50 mm, 1.3 μm column. The mobile phase composed of acetonitrile: 15 mM potassium dihydrogen orthophosphate (45 : 55 pumped isocratically at 0.4 mL/min. Detection wavelength was 262 nm in both methods. Results. The run time was 11 and 3 minutes for HPLC-DAD and UHPLC-UV assays, respectively. Both assays were linear (r2>0.999 with CV% and % error of the mean <3%. Limits of detection and quantitation were 0.82 and 2.73 μg/mL for HPLC-DAD and 1.04 and 3.16 μg/mL for UHPLC-UV, respectively. The methods quantitated PSZ in suspension dosage form with no observable interferences. Conclusions. Both assays were proven sensitive and selective according to ICH guidelines. UHPLC-UV assay exhibited some economic and chromatographic separation superiority.

  20. Rapid separation and identification of 54 major constituents in Buyang Huanwu decoction by ultra-fast HPLC system coupled with DAD-TOF/MS.

    Science.gov (United States)

    Liu, E-Hu; Qi, Lian-Wen; Peng, Yong-Bo; Cheng, Xiao-Lan; Wu, Qian; Li, Ping; Li, Chang-Yin

    2009-08-01

    Buyang Huanwu Decoction (BYHWD), is a well-known traditional Chinese preparation consisting of Radix Astragali, Radix Angelicae Sinensis, Rhizoma Ligustici Chuanxiong, Radix paeoniae Rubra, Flos Carthami, Semen Persicae and Lumbricus. An ultra-fast high-performance liquid chromatography (HPLC) method coupled with diode array detection (DAD) and electrospray ionization time-of-flight mass spectrometry (ESI-TOF/MS) has been developed for rapid separation and structural identification of constituents in BYHWD. Using an ultra-fast HPLC system with short columns (4.6 x 50 mm, 1.8 microm), the total analysis time for this complex prescription is less than 30 min. With various fragmentor voltages in TOF/MS, accurate mass measurements (less than 5 ppm error) for molecular ions and characteristic fragment ions could represent reliable identification criteria for these compounds. Fifty-four major constituents from BYHWD sample, including four C-glycosyl quinochalcones, four flavonoid O-glycosides, sixteen isoflavones, six monoterpene glycosides, eight saponins, four organic acids and five amino acids, were identified or tentatively characterized based on their retention times, DAD and TOF/MS data. All the compounds were further assigned in the seven individual crude drugs. In conclusion, the ultra-fast HPLC with DAD-TOF/MS is a highly useful and efficient technique to separate and identify constituents in complex matrices of herbal medicines or preparations.

  1. Chemometrics enhanced HPLC-DAD performance for rapid quantification of carbamazepine and phenobarbital in human serum samples.

    Science.gov (United States)

    Vosough, Maryam; Ghafghazi, Shiva; Sabetkasaei, Masoumeh

    2014-02-01

    This paper describes development and validation of a simple and efficient bioanalytical procedure for simultaneous determination of phenobarbital and carbamazepine in human serum samples using high performance liquid chromatography with photodiode-array detection (HPLC-DAD) regarding a fast elution methodology in less than 5 min. Briefly, this method consisted of a simple deproteinization step of serum samples followed by HPLC analysis on a Bonus-RP column using an isocratic mode of elution with acetonitrile/K2HPO4 (pH=7.5) buffer solution (45:55). Due to the presence of serum endogenous components as non-calibrated components in the sample, second-order calibration based on multivariate curve resolution-alternating least squares (MCR-ALS), has been applied on a set of absorbance matrices collected as a function of retention time and wavelengths. Acceptable resolution and quantification results were achieved in the presence of matrix interferences and the second-order advantage was fully exploited. The average recoveries for carbamazepine and phenobarbital were 89.7% and 86.1% and relative standard deviation values were lower than 9%. Additionally, computed elliptical joint confidence region (EJCR) confirmed the accuracy of the proposed method and indicated the absence of both constant and proportional errors in the predicted concentrations. The developed method enabled the determination of the analytes in different serum samples in the presence of overlapped profiles, while keeping experimental time and extraction steps at minimum. Finally, the serum concentration levels of carbamazepine in three time intervals were reported for morphine-dependents who had received carbamazepine for treating their neuropathic pain. © 2013 Elsevier B.V. All rights reserved.

  2. ¹H NMR and HPLC/DAD for Cannabis sativa L. chemotype distinction, extract profiling and specification.

    Science.gov (United States)

    Peschel, Wieland; Politi, Matteo

    2015-08-01

    The medicinal use of different chemovars and extracts of Cannabis sativa L. requires standardization beyond ∆9-tetrahydrocannabinol (THC) with complementing methods. We investigated the suitability of (1)H NMR key signals for distinction of four chemotypes measured in deuterated dimethylsulfoxide together with two new validated HPLC/DAD methods used for identification and extract profiling based on the main pattern of cannabinoids and other phenolics alongside the assayed content of THC, cannabidiol (CBD), cannabigerol (CBG) their acidic counterparts (THCA, CBDA, CBGA), cannabinol (CBN) and cannflavin A and B. Effects on cell viability (MTT assay, HeLa) were tested. The dominant cannabinoid pairs allowed chemotype recognition via assignment of selective proton signals and via HPLC even in cannabinoid-low extracts from the THC, CBD and CBG type. Substantial concentrations of cannabinoid acids in non-heated extracts suggest their consideration for total values in chemotype distinction and specifications of herbal drugs and extracts. Cannflavin A/B are extracted and detected together with cannabinoids but always subordinated, while other phenolics can be accumulated via fractionation and detected in a wide fingerprint but may equally serve as qualitative marker only. Cell viability reduction in HeLa was more determined by the total cannabinoid content than by the specific cannabinoid profile. Therefore the analysis and labeling of total cannabinoids together with the content of THC and 2-4 lead cannabinoids are considered essential. The suitability of analytical methods and the range of compound groups summarized in group and ratio markers are discussed regarding plant classification and pharmaceutical specification.

  3. Simultaneous quantification of delta-9-THC, THC-acid A, CBN and CBD in seized drugs using HPLC-DAD.

    Science.gov (United States)

    Ambach, Lars; Penitschka, Franziska; Broillet, Alain; König, Stefan; Weinmann, Wolfgang; Bernhard, Werner

    2014-10-01

    An HPLC-DAD method for the quantitative analysis of Δ(9)-tetrahydrocannabinol (THC), Δ(9)-tetrahydrocannabinolic acid-A (THCA-A), cannabidiol (CBD), and cannabinol (CBN) in confiscated cannabis products has been developed, fully validated and applied to analyse seized cannabis products. For determination of the THC content of plant material, this method combines quantitation of THCA-A, which is the inactive precursor of THC, and free THC. Plant material was dried, homogenized and extracted with methanol by ultrasonication. Chromatographic separation was achieved with a Waters Alliance 2695 HPLC equipped with a Merck LiChrospher 60 RP-Select B (5μm) precolumn and a Merck LiChroCart 125-4 LiChrospher 60 RP-Select B (5μm) analytical column. Analytes were detected and quantified using a Waters 2996 photo diode array detector. This method has been accepted by the public authorities of Switzerland (Bundesamt für Gesundheit, Federal Office of Public Health), and has been used to analyse 9092 samples since 2000. Since no thermal decarboxylation of THCA-A occurs, the method is highly reproducible for different cannabis materials. Two calibration ranges are used, a lower one for THC, CBN and CBD, and a higher one for THCA-A, due to its dominant presence in fresh plant material. As provider of the Swiss proficiency test, the robustness of this method has been tested over several years, and homogeneity tests even in the low calibration range (1%) show high precision (RSD≤4.3%, except CBD) and accuracy (bias≤4.1%, except CBN).

  4. Reversed Phase HPLC-DAD Profiling of Carotenoids, Chlorophylls and Phenolic Compounds in Adiantum capillus-veneris Leaves

    Directory of Open Access Journals (Sweden)

    Alam Zeb

    2017-04-01

    Full Text Available Adiantum capillus-veneris is important endangered fern species with several medicinal properties. In this study, the leaves samples were extracted and separated using reversed phase HPLC with DAD for carotenoids, chlorophylls and phenolic compounds. Separation of carotenoids and chlorophylls were carried out using a tertiary gradient system of water, MTBE and methanol-water, while a binary gradient system of methanol-water-acetic acid was used for phenolic profiling. Results revealed eight carotenoids, four pheophytins, and two chlorophylls. Lutein (806.0 μg/g, chlorophyll b′ (410.0 μg/g, chlorophyll a (162.4 μg/g, 9′-Z-neoxanthin (142.8 μg/g and all-E-violaxanthin (82.2 μg/g were present in higher amounts. The relatively high amounts of lutein may be one of the key indicator of beneficial antioxidant properties. The phenolic profile revealed a total of 13 compounds, namely 4-hydroxybenzoic acid, chlorogenic acid, caftaric acid, kaempferol glycosides, p-coumaric acid, rosmarinic acid, 5-caffeoylquinic acid, and quercetin glycosides. Kaempferol-3-sophorotrioside (58.7 mg/g, chlorogenic acid (28.5 mg/g, 5-O-caffeoylquinic acid (18.7 mg/g, coumaric acid (11.2 mg/g, and its derivative (33.1 mg/g were present in high amounts. These results suggest that the reversed phase HPLC profiling of Adiantum leaves provides a better understanding in to the actual composition of bioactive compounds, which may be responsible for the potential medicinal properties. Adiantum leaves rich in important bioactive phytochemicals can be used as a possible source of nutraceuticals or as a functional food ingredient.

  5. Reversed Phase HPLC-DAD Profiling of Carotenoids, Chlorophylls and Phenolic Compounds in Adiantum capillus-veneris Leaves

    Science.gov (United States)

    Zeb, Alam; Ullah, Fareed

    2017-04-01

    Adiantum capillus-veneris is important endangered fern species with several medicinal properties. In this study, the leaves samples were extracted and separated using reversed phase HPLC with DAD for carotenoids, chlorophylls and phenolic compounds. Separation of carotenoids and chlorophylls were carried out using a tertiary gradient system of water, MTBE and methanol-water, while a binary gradient system of methanol-water-acetic acid was used for phenolic profiling. Results revealed eight carotenoids, four pheophytins and two chlorophylls. Lutein (806.0 µg/g), chlorophyll b' (410.0 µg/g), chlorophyll a (162.4 µg/g), 9'-Z-neoxanthin (142.8 µg/g) and all-E-violaxanthin (82.2 µg/g)) were present in higher amounts. The relatively high amounts of lutein may be one of the key indicator of beneficial antioxidant properties. The phenolic profile revealed a total of thirteen compounds, namely p-hydroxybenzoic acid, chlorogenic acid, caftaric acid, kaempferol glycosides, p-coumaric acid, rosmarinic acid, 5-caffeoylquinic acid, and quercetin glycosides. Kaempferol-3-sophorotrioside (58.7 mg/g), chlorogenic acid (28.5 mg/g), 5-O-caffeoylquinic acid (18.7 mg/g), coumaric acid (11.2 mg/g) and its derivative (33.1 mg/g) were present in high amounts. These results suggest that the reversed phase HPLC profiling of adiantum leaves provides a better understanding in to the actual composition of bioactive compounds, which may be responsible for possible medicinal properties. Adiantum leaves rich in important bioactive phytochemicals can be used as a potential source of nutraceuticals or as a functional food ingredient.

  6. Complete separation of urinary metabolites of xylene in HPLC/DAD using β-cyclodextrin: application for biological monitoring.

    Science.gov (United States)

    Caporossi, Lidia; De Rosa, Mariangela; Papaleo, Bruno

    2010-10-01

    To determine the biomarkers of exposure to xylene, urinary 2-, 3- and 4-methyl-hippuric acids, a new HPLC/DAD analytical method has been developed, which uses β-cyclodextrin as an additive for elution; its complexing abilities are exploited to achieve complete chromatographic separation of the three isomers. The mobile phase was a 3% aqueous solution of β-cyclodextrin, pH 3, and methanol, 80:20, in isocratic conditions, with a flow rate of 1 mL/min. To optimize quantitative analysis three wavelengths were employed for detection: λ=198 nm, λ=200 nm, and λ=202 nm. SPE was applied for the extraction from urine samples of analytes. Validation parameters show recoveries always above 82%; LOD was set at 1 μg/mL with an LOQ of 3 μg/mL. The linear dynamic range (from 4 to 100 μg/mL) showed excellent correspondence. This method is rapid and inexpensive and can be applied to several samples simultaneously using a manifold for SPE extraction. The analytes were separated completely and could be fully quantified. The method was used for the analysis of urine samples from 54 workers exposed to xylene in hospital laboratories and showed a good applicability while allowing quantification even at low doses. Copyright © 2009 Elsevier B.V. All rights reserved.

  7. In Vitro Anticariogenic Effects of Drymocallis rupestris Extracts and Their Quality Evaluation by HPLC-DAD-MS3 Analysis

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    Sebastian Granica

    2013-07-01

    Full Text Available In this study, for the first time, we investigated in vitro inhibitory effects of Drymocallis rupestris extracts and their subfractions obtained with solvents of different polarity (aqueous, 50% ethanolic, diethyl ether, ethyl acetate and n-butanolic against bacterial viability and caries virulence factors of Streptococcus spp. strains. The diethyl ether subfraction (PRU2 showed bacteriostatic and bactericidal activity against mutans streptococci, with minimum inhibitory concentrations (MICs in the range of 0.75–1.5 mg/mL and minimum bactericidal concentrations (MBCs in the range of 1.5–3 mg/mL. Furthermore, PRU2 inhibited biofilm formation by Streptococci in a dose-dependent manner. It was also found that all five D. rupestris preparations exhibited diverse inhibitory effects on de novo synthesis of water-insoluble and water-soluble α-d-glucans by glucosyltransferases of the mutans group streptococci. The phytochemical profile of investigated samples was determined by spectrophotometric and chromatographic (HPLC-DAD-MS3 methods. The high polyphenol (total phenol, phenolic acids, tannins, proantocyanidins, and flavonoids contents were found which correlated with anticariogenic activity of the analyzed samples. The results demonstrate that D. rupestris extracts and their subfractions could become useful supplements for pharmaceutical products as a new anticariogenic agent in a wide range of oral care products. Further studies are necessary to clarify which phytoconstituents of D. rupestris are responsible for anticaries properties.

  8. RP-HPLC-DAD method for the determination of phenylepherine, paracetamol, caffeine and chlorpheniramine in bulk and marketed formulation

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    A.P. Dewani

    2014-11-01

    Full Text Available A simple, specific and accurate isocratic RP-HPLC-DAD method was developed for the simultaneous determination of phenylephrine, paracetamol, caffeine and chlorpheniramine in bulk and tablet dosage form. The four contents are present in variable concentrations and have variable chromatographic behavior making the process of analysis very difficult. For present studies a reversed-phase C-18 column (150 mm × 4.5 mm i.d., particle size 5 μm with mobile phase consisting of acetonitrile, methanol and 10 Mm phosphate buffer 16:22:62 (v/v (pH of buffer 2.5 ± 0.02, adjusted with ortho phosphoric acid was used. The flow rate was 1.0 ml/min and eluents were monitored at 280 nm. The mean retention times of phenylephrine, paracetamol, caffeine and chlorpheniramine were found to be 1.8, 3.1, 5.2 and 10.9 min, respectively. The method was validated in terms of linearity, range, specificity, accuracy, precision and robustness. The proposed method was successfully applied to the estimation of phenylephrine, paracetamol, caffeine and chlorpheniramine in combined tablet dosage form.

  9. Phlorotannin Extracts from Fucales Characterized by HPLC-DAD-ESI-MSn: Approaches to Hyaluronidase Inhibitory Capacity and Antioxidant Properties

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    Patrícia Valentão

    2012-12-01

    Full Text Available Purified phlorotannin extracts from four brown seaweeds (Cystoseira nodicaulis (Withering M. Roberts, Cystoseira tamariscifolia (Hudson Papenfuss, Cystoseira usneoides (Linnaeus M. Roberts and Fucus spiralis Linnaeus, were characterized by HPLC-DAD-ESI-MSn. Fucophloroethol, fucodiphloroethol, fucotriphloroethol, 7-phloroeckol, phlorofucofuroeckol and bieckol/dieckol were identified. The antioxidant activity and the hyaluronidase (HAase inhibitory capacity exhibited by the extracts were also assessed. A correlation between the extracts activity and their chemical composition was established. F. spiralis, the species presenting higher molecular weight phlorotannins, generally displayed the strongest lipid peroxidation inhibitory activity (IC50 = 2.32 mg/mL dry weight and the strongest HAase inhibitory capacity (IC50 = 0.73 mg/mL dry weight. As for superoxide radical scavenging, C. nodicaulis was the most efficient species (IC50 = 0.93 mg/mL dry weight, followed by F. spiralis (IC50 = 1.30 mg/mL dry weight. These results show that purified phlorotannin extracts have potent capabilities for preventing and slowing down the skin aging process, which is mainly associated with free radical damage and with the reduction of hyaluronic acid concentration, characteristic of the process.

  10. A novel application of microwave-assisted extraction of polyphenols from brewer's spent grain with HPLC-DAD-MS analysis.

    Science.gov (United States)

    Moreira, Manuela M; Morais, Simone; Barros, Aquiles A; Delerue-Matos, Cristina; Guido, Luís F

    2012-05-01

    This paper reports a novel application of microwave-assisted extraction (MAE) of polyphenols from brewer's spent grains (BSG). A 2(4) orthogonal composite design was used to obtain the optimal conditions of MAE. The influence of the MAE operational parameters (extraction time, temperature, solvent volume and stirring speed) on the extraction yield of ferulic acid was investigated through response surface methodology. The results showed that the optimal conditions were 15 min extraction time, 100 °C extraction temperature, 20 mL of solvent, and maximum stirring speed. Under these conditions, the yield of ferulic acid was 1.31 ± 0.04% (w/w), which was fivefold higher than that obtained with conventional solid-liquid extraction techniques. The developed new extraction method considerably reduces extraction time, energy and solvent consumption, while generating fewer wastes. HPLC-DAD-MS analysis indicated that other hydroxycinnamic acids and several ferulic acid dehydrodimers, as well as one dehydrotrimer were also present, confirming that BSG is a valuable source of antioxidant compounds.

  11. Comparison of Active Ingredient Content of Nanwuweizi and Beiwuweizi Collected in Different Seasons Detected by HPLC-DAD

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    Feng YAO

    2015-12-01

    Full Text Available Abstract Objective: To compare the active ingredient content of Nanwuweizi (Kadsura longepedunculata and Beiwuweizi (Fructus Schisandrae Chinensis collected in different seasons. Methods: Microwave-assisted extraction was applied to extract the active ingredients in Nanwuweizi and Beiwuweizi, with methanol as the solvent. High performance liquid chromatography-diode array detection ℃; mobile phase: water (A - acetonitrile (B; gradient elution: 0.8 mL/min; and detection wavelength: 225 nm. (HPLC-DAD method was used to analyze the content of each ingredient, with chromatographic conditions as follows: Brava BDS C18 column (250 mm×4.6 mm, 5 μm; column temperature: 30 Results: All ingredients waiting for detection reached their peaks within 75 min, with excellent degree of separation and higher number of theoretical plates. Standard curves were drawn, and the mean value of phase relationship of each ingredient was >0.999. The content of active ingredients in lignans of Nanwuweizi and Beiwuweizi showed tendency of increase first and then decreased. In addition, all ingredients in Beiwuweizi reached their peaks in September while those in Nanwuweizi in August. Conclusion: This method can be used to detect the content of multiple ingredients in Wuweizi (Schisandra chinensis simultaneously because it is simple in operation, precise in results and excellent in repeatability.

  12. Simultaneous Determination of Dexamethasone, Ondansetron, Granisetron, Tropisetron, and Azasetron in Infusion Samples by HPLC with DAD Detection

    Science.gov (United States)

    Chen, Fu-chao; Wang, Lin-hai; Guo, Jun; Shi, Xiao-ya

    2017-01-01

    A simple and rapid high-performance liquid chromatography with diode array detector (HPLC-DAD) method has been developed and validated for simultaneous quantification of five antiemetic agents in infusion samples: dexamethasone, ondansetron, granisetron, tropisetron, and azasetron. The chromatographic separation was achieved on a Phenomenex C18 column (4.6 mm × 150 mm, 5 μm) using acetonitrile-50 mM KH2PO4 buffer-triethylamine (25 : 74 : 1; v/v; pH 4.0). Flow rate was 1.0 mL/min with a column temperature of 30°C. Validation of the method was made in terms of specificity, linearity, accuracy, and intra- and interday precision, as well as quantification and detection limits. The developed method can be used in the laboratory to routinely quantify dexamethasone, ondansetron, granisetron, tropisetron, and azasetron simultaneously and to evaluate the physicochemical stability of referred drugs in mixtures for endovenous use. PMID:28168082

  13. Qualitative HPLC-DAD/ESI-TOF-MS Analysis, Cytotoxic, and Apoptotic Effects of Croatian Endemic Centaurea ragusina L. Aqueous Extracts.

    Science.gov (United States)

    Radan, Mila; Carev, Ivana; Tešević, Vele; Politeo, Olivera; Čulić, Vedrana Čikeš

    2017-09-01

    Centaurea ragusina L., an endemic Croatian plant species, revealed a good cytotoxic activity of aqueous extracts (AE) on human bladder (T24) and human glioblastoma (A1235) cancer cell lines. The chemical constituents were tentatively identified using high performance liquid chromatography HPLC-DAD/ESI-TOF-MS in negative ionization mode. The main compounds of herba extract were sesquiterpene lactones: solstitialin A 3,13-diacetate and epoxyrepdiolide; organic acid: quinic acid. The main compounds of flower extract were organic acids: quinic acid, citric acid, and malic acid; sesquiterpene lactone: cynaropicrin; phenolic compounds: chlorogenic acid and phenylpropanoid: syringin. The AE of C. ragusina were investigated for correlation of their effects on human bladder (T24) and human glioblastoma (A1235) cancer cell lines using the MTT assay. Although both extracts showed significant dose- and time-dependent cytotoxic activity against both cancer cell lines, the flower extract exhibited slightly higher activity. In order to determine type of cell death induced by treatment, cell lines were exposed subsequently to a treatment with both flower and herba AE. The majority of the cells died by induced apoptosis treatment. Flower AE (26.25%), compared to a leaf AE (22.15%) showed slightly higher percentage of an apoptosis in T24 cells, when compared to a non-treated cells (0.04%). © 2017 Wiley-VHCA AG, Zurich, Switzerland.

  14. Quantitative and qualitative analysis of common peaks in chemical fingerprint of Yuanhu Zhitong tablet by HPLC-DAD-MS/MS

    Institute of Scientific and Technical Information of China (English)

    Dao-Quan Tang; Xiao-Xiao Zheng; Xu Chen; Dong-Zhi Yang; Qian Du

    2014-01-01

    A quality control (QC) strategy for quantitative and qualitative analysis of “common peaks” in chemical fingerprint was proposed to analyze Yuanhu Zhitong tablet (YZT), using high performance liquid chromatography with diode array detector and tandem mass spectrometry (HPLC-DAD-MS/MS). The chromatographic separation was achieved on an Agilent Eclipse plus C18 column with a gradient elution using a mixture of 0.4‰ ammonium acetate aqueous (pH 6.0 adjusted with glacial acetic acid) and acetonitrile. In chemical fingerprint, 40 peaks were assigned as the “common peaks”. For quantification of “common peaks”, the detection wavelength was set at 254 nm, 270 nm, 280 nm and 345 nm, respectively. The method was validated and good results were obtained to simultaneously determine 10 analytes (protopine, jatrorrhizine, coptisine, palmatine, berberine, xanthotoxin, bergapten, tetrahydropalmatine, imperatorin and isoimperatorin). For qualification of “common peaks”, 33 compounds including 10 quantitative analytes were identified or tentatively characterized using LC-MS/MS. These results demonstrated that the present approach may be a powerful and useful tool to tackle the complex quality issue of YZT.

  15. Nutritional Composition and Antioxidant Capacity in Edible Flowers: Characterisation of Phenolic Compounds by HPLC-DAD-ESI/MSn

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    Inmaculada Navarro-González

    2014-12-01

    Full Text Available Edible flowers are commonly used in human nutrition and their consumption has increased in recent years. The aim of this study was to ascertain the nutritional composition and the content and profile of phenolic compounds of three edible flowers, monks cress (Tropaeolum majus, marigold (Tagetes erecta and paracress (Spilanthes oleracea, and to determine the relationship between the presence of phenolic compounds and the antioxidant capacity. Proximate composition, total dietary fibre (TDF and minerals were analysed according to official methods: total phenolic compounds (TPC were determined with Folin-Ciocalteu’s reagent, whereas antioxidant capacity was evaluated using Trolox Equivalent Antioxidant Capacity (TEAC and Oxygen Radical Absorbance Capacity (ORAC assays. In addition, phenolic compounds were characterised by HPLC-DAD-MSn. In relation to the nutritional value, the edible flowers had a composition similar to that of other plant foods, with a high water and TDF content, low protein content and very low proportion of total fat—showing significant differences among samples. The levels of TPC compounds and the antioxidant capacity were significantly higher in T. erecta, followed by S. oleracea and T. majus. Thirty-nine different phenolic compounds were tentatively identified, with flavonols being the major compounds detected in all samples, followed by anthocyanins and hydroxycynnamic acid derivatives. In T. erecta small proportions of gallotannin and ellagic acid were also identified.

  16. A rapid, one step preparation for measuring selected free plus SO2-bound wine carbonyls by HPLC-DAD/MS.

    Science.gov (United States)

    Han, Guomin; Wang, Hua; Webb, Michael R; Waterhouse, Andrew L

    2015-03-01

    Carbonyl compounds are produced during fermentation and chemical oxidation during wine making and aging, and they are important to wine flavor and color stability. Since wine also contains these compounds as α-hydroxysulfonates as a result of their reaction with sulfur dioxide, an alkaline pre-treatment requiring oxygen exclusion has been used to release these bound carbonyls for analysis. By modifying the method to hydrolyze the hydroxysulfonates with heating and acid in the presence of 2,4-dinitrophenylhydrazine (DNPH), the carbonyl compounds are simultaneously and quickly released and derivatized, resulting in a simpler and more rapid method. In addition, the method avoids air exclusion complications during hydrolysis by the addition of sulfur dioxide. The method was optimized for temperature, reaction time, and the concentrations of DNPH, sulfur dioxide and acid. The hydrazones were shown to be stable for 10 h, adequate time for chromatographic analysis by HPLC-DAD/MS. This method is demonstrated for 2-ketoglutaric acid, pyruvic acid, acetoin and acetaldehyde, wine carbonyls of very different reactivities, and it offers good specificity, high recovery and low limits of detection. This new rapid, simple method is demonstrated for the measurement of carbonyl compounds in a range of wines of different ages and grape varieties. Copyright © 2014 Elsevier B.V. All rights reserved.

  17. Nutritional composition and antioxidant capacity in edible flowers: characterisation of phenolic compounds by HPLC-DAD-ESI/MSn.

    Science.gov (United States)

    Navarro-González, Inmaculada; González-Barrio, Rocío; García-Valverde, Verónica; Bautista-Ortín, Ana Belén; Periago, María Jesús

    2014-12-31

    Edible flowers are commonly used in human nutrition and their consumption has increased in recent years. The aim of this study was to ascertain the nutritional composition and the content and profile of phenolic compounds of three edible flowers, monks cress (Tropaeolum majus), marigold (Tagetes erecta) and paracress (Spilanthes oleracea), and to determine the relationship between the presence of phenolic compounds and the antioxidant capacity. Proximate composition, total dietary fibre (TDF) and minerals were analysed according to official methods: total phenolic compounds (TPC) were determined with Folin-Ciocalteu's reagent, whereas antioxidant capacity was evaluated using Trolox Equivalent Antioxidant Capacity (TEAC) and Oxygen Radical Absorbance Capacity (ORAC) assays. In addition, phenolic compounds were characterised by HPLC-DAD-MSn. In relation to the nutritional value, the edible flowers had a composition similar to that of other plant foods, with a high water and TDF content, low protein content and very low proportion of total fat-showing significant differences among samples. The levels of TPC compounds and the antioxidant capacity were significantly higher in T. erecta, followed by S. oleracea and T. majus. Thirty-nine different phenolic compounds were tentatively identified, with flavonols being the major compounds detected in all samples, followed by anthocyanins and hydroxycynnamic acid derivatives. In T. erecta small proportions of gallotannin and ellagic acid were also identified.

  18. Rapid and quantitative determination of 10 major active components in Lonicera japonica Thunb. by ultrahigh pressure extraction-HPLC/DAD

    Science.gov (United States)

    Fan, Li; Lin, Changhu; Duan, Wenjuan; Wang, Xiao; Liu, Jianhua; Liu, Feng

    2015-01-01

    An ultrahigh pressure extraction (UPE)-high performance liquid chromatography (HPLC)/diode array detector (DAD) method was established to evaluate the quality of Lonicera japonica Thunb. Ten active components, including neochlorogenic acid, chlorogenic acid, 4-dicaffeoylquinic acid, caffeic acid, rutin, luteoloside, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C, and quercetin, were qualitatively evaluated and quantitatively determined. Scanning electron microscope images elucidated the bud surface microstructure and extraction mechanism. The optimal extraction conditions of the UPE were 60% methanol solution, 400 MPa of extraction pressure, 3 min of extraction time, and 1:30 (g/mL) solid:liquid ratio. Under the optimized conditions, the total extraction yield of 10 active components was 57.62 mg/g. All the components showed good linearity (r2 ≥ 0.9994) and recoveries. This method was successfully applied to quantify 10 components in 22 batches of L. japonica samples from different areas. Compared with heat reflux extraction and ultrasonic-assisted extraction, UPE can be considered as an alternative extraction technique for fast extraction of active ingredient from L. japonica.

  19. Simultaneous determination of potassium guaiacolsulfonate, guaifenesin, diphenhydramine HCl and carbetapentane citrate in syrups by using HPLC-DAD coupled with partial least squares multivariate calibration.

    Science.gov (United States)

    Dönmez, Ozlem Aksu; Aşçi, Bürge; Bozdoğan, Abdürrezzak; Sungur, Sidika

    2011-02-15

    A simple and rapid analytical procedure was proposed for the determination of chromatographic peaks by means of partial least squares multivariate calibration (PLS) of high-performance liquid chromatography with diode array detection (HPLC-DAD). The method is exemplified with analysis of quaternary mixtures of potassium guaiacolsulfonate (PG), guaifenesin (GU), diphenhydramine HCI (DP) and carbetapentane citrate (CP) in syrup preparations. In this method, the area does not need to be directly measured and predictions are more accurate. Though the chromatographic and spectral peaks of the analytes were heavily overlapped and interferents coeluted with the compounds studied, good recoveries of analytes could be obtained with HPLC-DAD coupled with PLS calibration. This method was tested by analyzing the synthetic mixture of PG, GU, DP and CP. As a comparison method, a classsical HPLC method was used. The proposed methods were applied to syrups samples containing four drugs and the obtained results were statistically compared with each other. Finally, the main advantage of HPLC-PLS method over the classical HPLC method tried to emphasized as the using of simple mobile phase, shorter analysis time and no use of internal standard and gradient elution. Copyright © 2010 Elsevier B.V. All rights reserved.

  20. Carotenoids from Foods of Plant, Animal and Marine Origin: An Efficient HPLC-DAD Separation Method

    Science.gov (United States)

    Strati, Irini F.; Sinanoglou, Vassilia J.; Kora, Lintita; Miniadis-Meimaroglou, Sofia; Oreopoulou, Vassiliki

    2012-01-01

    Carotenoids are important antioxidant compounds, present in many foods of plant, animal and marine origin. The aim of the present study was to describe the carotenoid composition of tomato waste, prawn muscle and cephalothorax and avian (duck and goose) egg yolks through the use of a modified gradient elution HPLC method with a C30 reversed-phase column for the efficient separation and analysis of carotenoids and their cis-isomers. Elution time was reduced from 60 to 45 min without affecting the separation efficiency. All-trans lycopene predominated in tomato waste, followed by all-trans-β-carotene, 13-cis-lutein and all-trans lutein, while minor amounts of 9-cis-lutein, 13-cis-β-carotene and 9-cis-β-carotene were also detected. Considering the above findings, tomato waste is confirmed to be an excellent source of recovering carotenoids, especially all-trans lycopene, for commercial use. Xanthophylls were the major carotenoids of avian egg yolks, all-trans lutein and all-trans zeaxanthin in duck and goose egg yolk, respectively. In the Penaeus kerathurus prawn, several carotenoids (zeaxanthin, all-trans-lutein, canthaxanthin, cryptoxanthin, optical and geometrical astaxanthin isomers) were identified in considerable amounts by the same method. A major advantage of this HPLC method was the efficient separation of carotenoids and their cis-isomers, originating from a wide range of matrices.

  1. Applications of a DAD-HPLC method for determination of loratadine on biological samples

    Directory of Open Access Journals (Sweden)

    Pavalache Georgeta

    2015-06-01

    Full Text Available The aim of research is to assess the active substance by a HPLC method for the separation and quantitative determination of loratadine. The method has been developed and validated on the standard solutions, in previous research. The current study was undertaken to present the results obtained from loratadine determination in biological samples (human serum, urine and breast milk. These results may be applicable on patients with different physiological conditions (aging, pregnancy or recently giving birth, etc. and pathological conditions which may interfere with the metabolism of loratadine. The used HPLC method detected loratadine concentrations in human serum samples, respectively urine samples, at 2 hours after drug administration. The method detected traces of loratadine which passed into breast milk, as well. Data were statistically interpreted using MED CALC 10.2 software. These results show that the applied method can be used for quantitative analysis of loratadine in biological fluids (all permissible limits of quality specifications being in the range 95- 105%.

  2. Computer-aided method for identification of major flavone/flavonol glycosides by high-performance liquid chromatography-diode array detection-tandem mass spectrometry (HPLC-DAD-MS/MS).

    Science.gov (United States)

    Wang, Zhengfang; Lin, Longze; Harnly, James M; Harrington, Peter de B; Chen, Pei

    2014-11-01

    A new computational tool is proposed here for tentatively identifying major (UV quantifiable) flavone/flavonol glycoside peaks of high performance liquid chromatogram (HPLC)-diode array detection (DAD)-tandem mass spectrometry (MS/MS) profiles based on a MATLAB-based script implementing an in-house algorithm. The HPLC-DAD-MS/MS profiles of red onion, Chinese lettuce, carrot leaf, and celery seed extracts were analyzed by the proposed computer-aided screening method for identifying possible flavone/flavonol glycoside peaks from the HPLC-UV and MS total ion current (TIC) chromatograms. The number of identified flavone/flavonol glycoside peaks of the HPLC-UV chromatograms is four, four, six, and nine for red onion, Chinese lettuce, carrot leaf, and celery seed, respectively. These results have been validated by human(s) experts. For the batch processing of nine HPLC-DAD-MS/MS profiles of celery seed extract, the entire script execution time was within 15 s while manual calculation of only one HPLC-DAD-MS/MS profile by a flavonoid expert could take hours. Therefore, this MATLAB-based screening method is able to facilitate the HPLC-DAD-MS/MS analysis of flavone/flavonol glycosides in plants to a large extent.

  3. A Comparative Study of Newly Developed HPLC-DAD and UHPLC-UV Assays for the Determination of Posaconazole in Bulk Powder and Suspension Dosage Form.

    Science.gov (United States)

    Hamdy, Dalia A; Belal, Tarek S

    2014-01-01

    Objective. To develop and compare HPLC-DAD and UHPLC-UV assays for the quantitation of posaconazole in bulk powder and suspension dosage form. Methods. Posaconazole linearity range was 5-50 μg/mL for both assays. For HPLC-DAD assay, samples were injected through Zorbax SB-C18 (4.6 × 250 mm, 5 μm) column. The gradient elution composed of the mobile phase acetonitrile: 15 mM potassium dihydrogen orthophosphate (30 : 70 to 80 : 20, linear over 7 minutes) pumped at 1.5 mL/min. For UHPLC-UV assay, samples were injected through Kinetex-C18 (2.1 × 50 mm, 1.3 μm) column. The mobile phase composed of acetonitrile: 15 mM potassium dihydrogen orthophosphate (45 : 55) pumped isocratically at 0.4 mL/min. Detection wavelength was 262 nm in both methods. Results. The run time was 11 and 3 minutes for HPLC-DAD and UHPLC-UV assays, respectively. Both assays were linear (r (2) > 0.999) with CV% and % error of the mean DAD and 1.04 and 3.16 μg/mL for UHPLC-UV, respectively. The methods quantitated PSZ in suspension dosage form with no observable interferences. Conclusions. Both assays were proven sensitive and selective according to ICH guidelines. UHPLC-UV assay exhibited some economic and chromatographic separation superiority.

  4. Extraction optimization of coumarins from radix angelicae pubescentis by HPLC-DAD coupled with uniform design

    Institute of Scientific and Technical Information of China (English)

    GUO Fang-qiu; HUANG Lan-fang; ZHOU Huo-fei; JIANG Qiu-ju

    2006-01-01

    Uniform design was used to optimize extraction condition of direct refluence extraction of coumarins from the Chinese traditional medicine of radix angelicae pubescentis(Duhuo); the sum peak area of coumarins separated with high performance liquid chromatography(HPLC) at detection wavelength of 320 nm was considered as detection index, two factors of solvent concentration and extraction time were mainly studied at extraction temperature of 85 ℃ and a volume ratio of solvent to sample of 10: 1. Optimal subclass, quadric polynomial step by step aggression and neural network method were applied to process the experimental results. The results show that the first and second methods give the same factors combination (concentration of ethanol: 95 %, extraction time: 3.6 h) and the second method is much better than the first one. The extraction model is consequently developed.

  5. HPLC-DAD phenolic profile, cytotoxic and anti-kinetoplastidae activity of Melissa officinalis.

    Science.gov (United States)

    Cunha, Francisco; Tintino, Saulo R; Figueredo, Fernando; Barros, Luiz; Duarte, Antonia E; Vega Gomez, Maria Celeste; Coronel, Cathia Cecilia; Rolón, Mírian; Leite, Nadghia; Sobral-Souza, Celestina E; Brito, S V; Waczuc, Emily Pansera; Boligon, Aline Augusti; Athayde, Margareth; Kamdem, Jean Paul; Coutinho, Henrique Douglas Melo; Franco, Jéferson

    2016-09-01

    Context Melissa officinalis subsp. inodora Bornm. (Lamiaceae) has been used since ancient times in folk medicine against various diseases, but it has not been investigated against protozoa. Objective To evaluate the activities of M. officinalis against Leishmania braziliensis, Leishmania infantum and Trypanosoma cruzi as well as its cytotoxicity in fibroblast cell line. Materials and methods The fresh leaves were chopped into 1 cm(2) pieces, washed and macerated with 99.9% of ethanol for 72 h at room temperature. Antiparasitic activity of M. officinalis was accessed by direct counting of cells after serial dilution, while the cytotoxicity of M. officinalis was evaluated in fibroblast cell line (NCTC929) by measuring the reduction of resazurin. The test duration was 24 h. High-performance liquid chromatography (HPLC) was used to characterise the extract. Results The extract at concentrations of 250 and 125 μg/mL inhibited 80.39 and 54.27% of promastigote (LC50  value = 105.78 μg/mL) form of L. infantum, 80.59 and 68.61% of L. brasiliensis (LC50 value  = 110.69 μg/mL) and against epimastigote (LC50 value  = 245.23 μg/mL) forms of T. cruzi with an inhibition of 54.45 and 22.26%, respectively, was observed. The maximum toxicity was noted at 500 μg/mL with 95.41% (LC50  value = 141.01 μg/mL). The HPLC analysis identified caffeic acid and rutin as the major compounds. Discussion The inhibition of the parasites is considered clinically relevant (officinalis can be considered a potential alternative source of natural products with antileishmania and antitrypanosoma activities.

  6. Rapid profiling of phenolic compounds of green and fermented Bergenia crassifolia L. leaves by UPLC-DAD-QqQ-MS and HPLC-DAD-ESI-QTOF-MS.

    Science.gov (United States)

    Salminen, Juha-Pekka; Shikov, Alexander N; Karonen, Maarit; Pozharitskaya, Olga N; Kim, Jorma; Makarov, Valery G; Hiltunen, Raimo; Galambosi, Bertalan

    2014-01-01

    Bergenia crassifolia L., Saxifragaceae, is an evergreen perennial plant known in traditional medicine of Russia, Mongolia and China. Polyphenols are responsible for the number of pharmacological effects of Bergenia. UPLC-DAD-QqQ-MS and LC-DAD-ESI-QTOF-MS were used for the rapid profiling of phenolic compounds, mainly hydrolysable tannins. Green leaves consisted of 55% ellagitannins, 29% gallic acid derivatives and 11% flavonoids, with the remaining gallic acid, arbutin, bergenin and caffeoyl quinic acid. In fermented leaves, 31% of gallic acid was found, followed with 28% ellagitannins, 18% gallic acid derivatives and 18% flavonoids, with the remaining caffeoyl quinic acid, bergenin and arbutin. Tellimagrandin I, pedunculagin, caffeoyl quinic acid, monogalloyl quinic acid, 1-O-galloylglucose and 1,2,6-tri-O-galloylglucose were identified for the very first time.

  7. Qualitative analysis of Psoraleae Fructus by HPLC-DAD/TOF-MS fingerprint and quantitative analysis of multiple components by single marker.

    Science.gov (United States)

    Luan, Lianjun; Shen, Xiaoyu; Liu, Xuesong; Wu, Yongjiang; Tan, Manliang

    2017-08-04

    A variety of bioactive substances may account for the recognized efficacy and wide clinical application of Psoraleae Fructus in China. A high-performance liquid chromatography-diode array detector (HPLC-DAD) fingerprint method was developed to present the comprehensive phytochemical profile of the crude drug. Thirteen major compounds were separated and identified by HPLC coupled with time-of-flight mass spectrometry (HPLC/TOF-MS), namely psoralenoside (PO), isopsoralenoside (IPO), psoralen (PS), isopsoralen (IPS), neobavaisoflavone (NBF), bavachin (BC), corylin (CN), bavachromene (BCM), psoralidin (PD), isobavachalcone (IBC), bacachinin (BCN), corylifol A (CA) and bakuchiol (BK). Then quantitative analysis of multiple components by single marker (QAMS) was applied in content determination of PO, IPO, PS, IPS, BC, IBC, BCN, CA and BK, with NBF as the internal standard. The calculation results indicated no significant difference from the traditional external standard method (p > 0.05, RSD chemical compositions, especially when there is a shortage of reference substances. In conclusion, simultaneous qualitative and quantitative analysis of Psoraleae Fructus may be fulfilled through the newly proposed method of QAMS combined with HPLC-DAD/TOF-MS fingerprint. Copyright © 2017 John Wiley & Sons, Ltd.

  8. Spondias tuberosa (Anacardiaceae) leaves: profiling phenolic compounds by HPLC-DAD and LC-MS/MS and in vivo anti-inflammatory activity.

    Science.gov (United States)

    da Silva Siqueira, Emerson Michell; Félix-Silva, Juliana; de Araújo, Lorena Maria Lima; Fernandes, Julia Morais; Cabral, Bárbara; Gomes, Jacyra Antunes Dos Santos; de Araújo Roque, Alan; Tomaz, José Carlos; Lopes, Norberto Peporine; de Freitas Fernandes-Pedrosa, Matheus; Giordani, Raquel Brandt; Zucolotto, Silvana Maria

    2016-10-01

    Spondias tuberosa is a medicinal plant used by several local communities in northeast Brazil to treat infections, digestive disorders and inflammatory conditions. The study aimed to identify and quantify the major phenolic in hydroethanolic extract of leaves from S. tuberosa and to evaluate its anti-inflammatory potential. The chemical profile of extract was analyzed by HPLC-DAD and HPLC-MS. The in vivo anti-inflammatory activity was investigated in carrageenan-induced hind paw edema and peritonitis models in mice. Identified and quantified through HPLC-DAD or HPLC-MS analyses of S. tuberosa extract were the following compounds: chlorogenic acid, caffeic acid, rutin and isoquercitrin. The inflammatory response to carrageenan was significantly reduced in both models by S. tuberosa extract. In hind paw edema, the edematogenic response was reduced by up to 63.6% and the myeloperoxidase activity was completely inhibited. In the peritonitis model, the total cell migration into the peritoneal cavity was reduced by up to 65%. The results obtained give evidence of the anti-inflammatory action of S. tuberosa and suggest the potential therapeutic benefit of this plant on inflammatory conditions. The chlorogenic acid, caffeic acid, rutin and isoquercitrin identified and quantified in S. tuberosa leaves enable us to suggest that these compounds could be used as chemical markers for quality control of derivative products from this species. Copyright © 2016 John Wiley & Sons, Ltd.

  9. Separation of positional CPP isomers by chiral HPLC-DAD of seized tablets.

    Science.gov (United States)

    Schürenkamp, Jennifer; Beike, Justus; Pfeiffer, Heidi; Köhler, Helga

    2011-01-01

    Meta-chlorophenylpiperazine, one of the synthetic piperazine-derived designer drugs, is to date controlled as an illicit substance in five European member states. Depending on the position of the chlorine atom, different positional isomers of CPP (ortho-, meta- and para-) are possible. Therefore, there is a need to develop an analytical method for the separation and identification of the three 1-chlorophenylpiperazines in tablets containing CPP. In this work, the position isomers o-, m- and p-CPP were separated by liquid chromatography (HPLC) on a reversed-phase chiral column. Different mobile phase compositions and pH ranges were systematically studied to find optimum chromatographic conditions. Best results were achieved with isocratic mobile phase of triethyl amine buffer and methanol (V/V = 70/30) at pH 9 with a flow rate of 0.8 ml/min. The method was validated in terms of selectivity, linearity, limit of detection and quantification and precision. At last, the developed method was successfully applied on seized ecstasy tablets.

  10. An improved HPLC-DAD method for clavulanic acid quantification in fermentation broths of Streptomyces clavuligerus.

    Science.gov (United States)

    Ramirez-Malule, Howard; Junne, Stefan; López, Carlos; Zapata, Julian; Sáez, Alex; Neubauer, Peter; Rios-Estepa, Rigoberto

    2016-02-20

    Clavulanic acid (CA) is an important secondary metabolite commercially produced by cultivation of Streptomyces clavuligerus (Sc). It is a potent inhibitor of bacterial β-lactamases. In this work, a specific and improved high performance liquid chromatography (HPLC) method, using a C-18 reversed phase column, diode array detector and gradient elution for CA quantification in fermentation broths of Sc, was developed and successfully validated. Samples were imidazole-derivatized for the purpose of creating a stable chromophore (clavulanate-imidazole). The calibration curve was linear over a typical range of CA concentration between 0.2 and 400mg/L. The detection and quantification limits were 0.01 and 0.02mg/L, respectively. The precision of the method was evaluated for CA spiked into production media and a recovery of 103.8%, on average, was obtained. The clavulanate-imidazole complex was not stable when the samples were not cooled during the analysis. The recovery rate was 39.3% on average. This assay was successfully tested for CA quantification in samples from Sc fermentation, using both, a chemically defined and a complex medium.

  11. d-Phenothrin-induced oxidative DNA damage in rat liver and kidney determined by HPLC-ECD/DAD.

    Science.gov (United States)

    Atmaca, Enes; Aksoy, Abdurrahman

    2015-05-01

    The objective of this study was to assess the risk of genotoxicity of d-phenothrin by measuring the oxidative stress it causes in rat liver and kidney. The level of 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG)/10(6) 2'-deoxyguanosine (dG) was measured by using high performance liquid chromatography (HPLC) with a diode array (DAD) and an electrochemical detector (ECD). Sixty male Wistar albino rats were randomly divided into five experimental groups and one control group of 10 rats/group. d-phenothrin was administered intraperitoneally (IP) to the five experimental groups at 25 mg/kg (Group I), 50 mg/kg (Group II), 66.7 mg/kg (Group III), 100 mg/kg (Group IV), and 200 mg/kg (Group V) for 14 consecutive days, and the control group received only the vehicle, dimethyl sulfoxide (DMSO). DNA from samples frozen in liquid nitrogen was isolated with a DNA isolation kit. Following digestion with nuclease P1 and alkaline phosphatase (ALP), hydrolyzed DNA was subjected to HPLC. The dG and 8-oxodG levels were analyzed with a DAD and ECD, respectively. In the experimental groups, the mean 8-oxodG/10(6) dG levels were 48.15 ± 7.43, 68.92 ± 20.66, 82.07 ± 14.15, 85.08 ± 28.50, and 89.14 ± 21.73 in livers and 39.06 ± 7.63, 59.69 ± 14.22, 61.13 ± 17.46, 65.13 ± 23.40, and 72.66 ± 19.04 in kidneys of Groups I, II, III, IV, and V, respectively. The mean 8-oxodG/10(6) dG levels in the control groups were 44.96 ± 12.66 for the liver and 39.07 ± 4.80 for the kidney. A statistically significant (p < 0.05), dose-dependent increase in oxidative DNA damage was observed in both organs of animals exposed to d-phenothrin when compared to controls. Furthermore, the liver showed a significantly higher level of oxidative DNA damage than the kidney (p < 0.01). In conclusion, d-phenothrin administered to rats intraperitoneally for 14 consecutive days generated free radical species in a dose-dependent manner and caused oxidative

  12. Molecularly imprinted pipette-tip solid phase extraction for selective determination of fluoroquinolones in human urine using HPLC-DAD.

    Science.gov (United States)

    de Oliveira, Hanna Leijoto; da Silva Anacleto, Sara; da Silva, Anny Talita Maria; Pereira, Arnaldo César; de Souza Borges, Warley; Figueiredo, Eduardo Costa; Borges, Keyller Bastos

    2016-10-15

    A simple method using HPLC-DAD was developed for the determination of fluoroquinolones in human urine including ciprofloxacin (CIPRO), enrofloxacino (ENRO), marbofloxacino (MARBO) and norfloxacin (NOR). In addition, it was studied the extraction of fluoroquinolones in human urine samples using pipette tip-based molecularly imprinted polymers solid phase extraction (PT-MIPs-SPE). With the goal of finding the best procedure for extraction of four fluoroquinolones in human urine, several parameters that are likely to affect the efficiency of extraction during sample preparation, including the washing solvent, type and volume of eluent, amount of material, the volume of the sample, pH and the ionic strength were systematically optimized. Chromatographic separations of fluoroquinolones were hit within 10min using a Synergi(®) C18 (250×4.6mm, 4μm) column and mobile phase consisting of water (10mM of phosphoric acid, the pH adjusted at 3.29 with triethylamine) : acetonitrile (85.7: 14.3, v/v) at a flow rate of 1.5mLmin(-1). Detection was performed at 290nm. The average extraction recoveries/standard deviation relative to ENRO, CIPRO, NOR and MARBO were 96.40±5.51%, 42.47±4.81%, 41.82±7.99% and 87.49±4.70, respectively. The method was liner from 39 to 1260ngmL(-1) for each fluoroquinolone with correlation coefficient of 0.9904, 0.9910, 0.9914 and 0.9919, to ENRO, CIPRO, NOR and MARBO, respectively. The assays of within-day and between-day precision and accuracy for all analytes were studied at three concentration levels and were lower than 15%. The method was successfully employed in a preliminary cumulative urinary excretion study after administration of CIPRO to a healthy volunteer. Copyright © 2016 Elsevier B.V. All rights reserved.

  13. Optimization of extraction and determination of emodin from Polygonum cuspidatum Sieb. et Zucc. products by HPLC-DAD

    Institute of Scientific and Technical Information of China (English)

    LU Hong-mei; LIANG Yi-zeng; NI Wang-dong; MAN Rui-lin

    2006-01-01

    A uniform experimental design procedure was used to investigate the effects of some operating parameters on the extraction of emodin from Polygonum cuspidatum Sieb. et Zucc. products. Variables tested were volume ratio of material to solvent, size of material, extraction time and temperature and composition of extraction solvent (mixtures of acetone-water). Each variable was tested at seven levels; 7 experiments were performed in random order. Analyses of the extracts were performed by high-performance liquid chromatography with diode array detection(HPLC-DAD).Analytical responses were processed by using a forward regression analysis, in order to find polynomial function describing the relationship between variables and responses. For all the analytes the experimental conditions for providing the highest extraction yield inside the experimental domain considered were found. Finally, a simple, rapid and accurate analytical method was developed for the determination of emodin by high performance liquid chromatography. The separation is achieved within 25 min on an ODS column using methanol and water as gradient mobiles. Emodin can be quantified by using external standard method detecting at 436 nm. Good linearity is obtained with correlation coefficient exceeding 0.9986 and the detection limit and the quantification limit are 1.53 and 3.23 mg/L respectively. This method shows good reproducibility for the quantification of the emodin with intra-day and inter-day relative standard deviation less than 2.3% and 5.6% respectively. Under optimized extraction conditions, the recovery of the standard is 96.5%. The validated method is successfully applied to quantify the emodin in seven Polygonum cuspidatum sieb. Et zucc. products, which provided an idea for the pre-treatment of determination of active compounds in traditional Chinese medicines.

  14. HPLC-DAD Method for the Pharmacokinetic Interaction Study of Atorvastatin with Pioglitazone and Cholestyramine in Wistar Rats.

    Science.gov (United States)

    Sharma, Ritesh N; Pancholi, Shyam S

    2014-01-01

    Carotid intima-media thickness is used as a surrogate marker for cardiovascular complications in diabetes mellitus. The combination of atorvastatin and pioglitazone was found to be effective in reducing the thickness of the carotid intima-media layer. The method of RP-HPLC coupled with a diode array detector (DAD) was developed for the pharmacokinetic interaction study of atorvastatin with pioglitazone and cholestyramine, respectively, in Wistar rats. Atorvastatin (ATR) and pioglitazone (PIO) were resolved on a C18 column with a mobile phase composed of 48% methanol, 19% acetonitrile, and 33% 10 mM ammonium formate (v/v/v; pH 3.5±0.3, by formic acid) and a 260 nm detection wavelength on the diode array detector. The method was validated according to international standards with good reproducibility and linear response; mean (r) 0.9987 and 0.9972 to ATR and PIO, respectively. The coefficients of variation of intra- and interassay precision ranged between 4.95-8.12 and 7.29-9.67, respectively. Pharmacokinetic parameters were determined in rats following an oral administration of atorvastatin in the presence and absence of pioglitazone and also with cholestyramine. Compared with the control given atorvastatin alone, the Cmax and AUC of atorvastatin were merely unchanged in rats with the co-administration of pioglitazone, while they decreased by nearly 21 and 15%, respectively, with the concurrent use of cholestyramine. There were no significant changes in Tmax and the plasma half-life (T1/2 ) of atorvastatin in both cases. The performed experiment demonstrated that the presented method was suitable for the estimation and pharmacokinetic interaction study of atorvastatin with pioglitazone and cholestyramine in Wistar rat plasma.

  15. [Qualitative and quantitative analysis of major constituents of Paeoniae Radix Alba and Paeoniae Radix Rubra by HPLC-DAD-Q-TOF-MS/MS].

    Science.gov (United States)

    Liu, Jie; Chen, Lin; Fan, Cai-rong; Li, Huang; Huang, Ming-qing; Xiang, Qing; Xu, Wen; Xu, Wei; Chu, Ke-dan; Lin, Yu

    2015-05-01

    In order to explore the differences of chemical constituents of Paeoniae Radix Alba and Paeoniae Radix Rubra, a qualitative analytical method of liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (HPLC-Q-TOF-MS/MS) was developed for identification of multi-constituents and an HPLC-DAD analytical method was developed for simultaneously determining 14 major compounds (gallic acid, protocatechuic acid, paeoniflorin sulfonate, protocatechuic aldehyde, methyl gallate, oxypaeoniflorin, catechin, albiflorin, and paeoniflorin, ethyl gallate, benzoic acid, pentagaloylglucose, benzoyl-paeoniflorin, and paeonol) in Paeoniae Radix Alba and Paeoniae Radix Rubra. Q-TOF/MS qualitative analysis was performed under negative ion mode and inferred 38 components of Paeoniae Radix Alba and 30 components of Paeoniae Radix Rubra. HPLC-DAD quantitative method result showed the contents of 8 ingredients were different between Paeoniae Radix Alba and Paeoniae Radix Rubra. The results indicated that the new approach was applicable in qualitative and quantitative quality control of Paeoniae Radix Alba and Paeoniae Radix Rubra.

  16. An insight on the alkaloid content of Capparis spinosa L. root by HPLC-DAD-MS, MS/MS and (1)H qNMR.

    Science.gov (United States)

    Khatib, Mohamad; Pieraccini, Giuseppe; Innocenti, Marzia; Melani, Fabrizio; Mulinacci, Nadia

    2016-05-10

    The Capparis spinosa L. has a wide distribution in the Old World from South Europe, North and East Africa, Madagascar, Southwest and Central Asia to Australia and Oceania. The consolidated traditional use of C. spinosa root as remedy against different pains in human is well known since the antiquity. Various secondary metabolites have been found in caper plant, nevertheless, few studies have been focused to the analysis of root constituents. To date, several free and glycosilated spermidine alkaloids and a more polar alkaloid, the stachydrine, have been isolated from the root of C. spinosa. Aim of this work was to improve the knowledge on the alkaloid content of the root of a Syrian sample of C. spinosa by HPLC-DAD-MS(n) and to propose methods to quantify these molecules in different raw extracts. A decoction, an hydroalcoholic extraction and a fractionation process to selectively recover the spermidine alkaloids were applied. To our knowledge, this is the first HPLC-DAD-MS(n) profile that pointed out the co-presence of stachydrine, several isobaric forms of capparispine and/or capparisine in free and glycosylated forms and some isobars of isocodonocarpine or codonocarpine as monoglycosides in extracts of C. spinosa root. The determination by HPLC/DAD for the spermidine alkaloids expressed as p-OH-coumaric acid gave values up to 3.5mg/g dried root and the stachydrine evaluated by (1)H NMR was close to 12.5mg/g dried root. Overall, the total alkaloids were almost doubled in hydroalcoholic extract with respect to the decoction, and the stachydrine in the cortex was almost double than in the whole root.

  17. HPLC-DAD/MS/MS Analysis of Tea Polyphenols in Leaves of Baiyedancong and Huangguanyin%白叶单枞和黄观音茶树芽叶中茶多酚的HPLC-DAD/MS/MS分析

    Institute of Scientific and Technical Information of China (English)

    卢嘉丽; 王冬梅; 苗爱清; 杨得坡

    2007-01-01

    目的 对两个乌龙茶的代表茶树品种白叶单枞和黄观音芽叶中的茶多酚进行定性、定量比较分析.方法 茶样经体积分数50%乙腈室温超声提取后进行HPLC-DAD/MS/MS分析;采用ODS柱,流动相为乙腈和1%甲酸水溶液,进行线性梯度洗脱,扫描波长范围为200~700 nm;ESI离子化,正负离子同时检测,m/z扫描范围为150~1 500 amu,MS2碰撞能量为35 eV.结果 通过与8种儿茶素对照品的比较分析,在这两个茶树品种的芽叶中鉴定出了9种儿茶素类化合物,2种嘌呤生物碱以及2种非儿茶素类茶多酚,并测定了各儿茶素类化合物的含量,回收率在97.11%~99.73%之间.结论 两个茶树品种芽叶中的儿茶素类化合物组成种类基本相近,白叶单枞中酯型儿茶素的含量比黄观音高,而非酯型儿茶素的含量却比较低;(-)-epigallocatechin-3-(3"-O-methyl)gallate和(-)-epigallocatechin-3,5-digallate在我国茶树现有栽培品种中的发现为首次报道.

  18. Simultaneous Quantification of Limonin, Two Indolequinazoline Alkaloids, and Four Quinolone Alkaloids in Evodia rutaecarpa (Juss.) Benth by HPLC-DAD Method

    OpenAIRE

    Pei-ting Zhang; Bi-yan Pan; Qiong-feng Liao; Mei-cun Yao; Xin-jun Xu; Jin-zhi Wan; Dan Liu; Zhi-yong Xie

    2013-01-01

    A simple and efficient HPLC-DAD (225 nm) method was developed and validated for the simultaneous determination of limonin and six key alkaloids (evodiamine, rutaecarpine, 1-methyl-2-undecyl-4(1H)-quinolone, evocarpine, 1-methy-2-[(6Z,9Z)]-6,9-pentadecadienyl-4-(1H)-quinolone, and dihydroevocarpine) in Evodia rutaecarpa (Juss.) Benth, which has been widely used as one of the Traditional Chinese Medicines. The chromatographic separation was carried out on a Hypersil BDS C18 column, and gradient...

  19. Analysis of Metabolites of Bazhen Decoction in Rats by HPLC-DAD-ESI/MSn Technique%基于HPLC-DAD-ESL/MSn技术分析八珍汤在大鼠体内的代谢化学成分

    Institute of Scientific and Technical Information of China (English)

    李文兰; 陈奇; 白晶; 孙志; 杨洋; 季宇彬

    2011-01-01

    OBJECTIVE To identify the prototypes and metabolites of Bazhen Decoction in rat serum after oral administration.METHODS The chemical components of Bazhen Decoction in rat serum samples obtained after oral administration were analyzed by high performance liquid chromatography-electrospray ion trap mass spectrometry (HPLC-ESI-MS) with selected ion monitoring and secondary fragment ion monitoring on line.RFSULTS 22 Constituents, including 16 prototypes and 6 metabolites, were detected in setum samples.The prototypes were atractylenolide Ⅰ , 5-hudroxymethyl-2-furfural, hydroxypaeoniflorin, atractylenolide Ⅲ, albiflorin,paeoniflorin, liquiritin, ferulic acid, ligustilide, ginsenoside Rg1 and ginsenoside Rb1-.The metabolites were paeonimetabolin Ⅰ , 3-0-β-D-18β-glycyrrhetic acid monoglucuronide, glycyrrhetic acid, ginsenoside Rh1 and ginsenoside Rd.CONCLUSION This sudy reveals the material base of the therapeutic effect of Bazhen Decoction, and illustrates the principle of compatibility of traditional Chinese medicine.It also provides theoretic basis for establishing the quality standard of Bazhen Decoction.%目的 鉴定口服八珍汤后大鼠血中原型成分及代谢产物.方法 对八珍汤给药后所得血清样品,应用HPLC-DAD-MS联用技术,在线进行选择离子扫描(SIM)、二级碎片离子全扫描,对八珍汤的体内代谢化学成分进行解析.结果 大鼠口服八珍汤后从血中发现了22个入血成分,其中16个成分为八珍汤所含成分的原型,6个为新产生的代谢产物,其中原型成分分别为白术内酯I、5-羟甲基-2-糖酸、羟基芍药苷、白术内酯皿、芍药内酯苷、芍药苷、甘草苷、阿魏酸、藁本内酯、人参皂苷Rg1,、人参皂苷Rb1;代谢产物分别为芍药苷代谢素I、3-O-β-D-18β-甘草次酸单葡萄糖醛酸苷、甘草次酸、人参皂苷Rh1、人参皂苷Rd.结论 为揭示八珍汤的药效物质基础、阐明中药复方的配伍原则及质量标准制定提供科学的理论依据.

  20. Screening of Satureja subspicata Vis. Honey by HPLC-DAD, GC-FID/MS and UV/VIS: Prephenate Derivatives as Biomarkers.

    Science.gov (United States)

    Jerković, Igor; Kranjac, Marina; Marijanović, Zvonimir; Zekić, Marina; Radonić, Ani; Tuberoso, Carlo Ignazio Giovanni

    2016-03-21

    The samples of Satureja subspicata Vis. honey were confirmed to be unifloral by melissopalynological analysis with the characteristic pollen share from 36% to 71%. Bioprospecting of the samples was performed by HPLC-DAD, GC-FID/MS, and UV/VIS. Prephenate derivatives were shown to be dominant by the HPLC-DAD analysis, particularly phenylalanine (167.8 mg/kg) and methyl syringate (MSYR, 114.1 mg/kg), followed by tyrosine and benzoic acid. Higher amounts of MSYR (3-4 times) can be pointed out for distinguishing S. subspicata Vis. honey from other Satureja spp. honey types. GC-FID/MS analysis of ultrasonic solvent extracts of the samples revealed MSYR (46.68%, solvent pentane/Et2O 1:2 (v/v); 52.98%, solvent CH2Cl2) and minor abundance of other volatile prephenate derivatives, as well as higher aliphatic compounds characteristic of the comb environment. Two combined extracts (according to the solvents) of all samples were evaluated for their antioxidant properties by FRAP and DPPH assay; the combined extracts demonstrated higher activity (at lower concentrations) in comparison with the average honey sample. UV/VIS analysis of the samples was applied for determination of CIE Lab colour coordinates, total phenolics (425.38 mg GAE/kg), and antioxidant properties (4.26 mmol Fe(2+)/kg (FRAP assay) and 0.8 mmol TEAC/kg (DDPH assay)).

  1. Screening and quantitative analysis of antioxidants in the fruits of Livistona chinensis R. Br using HPLC-DAD-ESI/MS coupled with pre-column DPPH assay.

    Science.gov (United States)

    Yao, Hong; Chen, Yan; Shi, Peiying; Hu, Juan; Li, Shaoguang; Huang, Liying; Lin, Jianhua; Lin, Xinhua

    2012-12-15

    In this study, a high performance liquid chromatography-photo diode array detection-electrospray ionization tandem mass spectrometry (HPLC-DAD/ESI-MS) with pre-column DPPH assay is developed for screening the antioxidant components in the fruits of Livistona chinensis R. Br. Accordingly, six antioxidative flavonoids are identified as orientin, isoorientin, vitexin, isovitexin, isorhamnetin-3-O-glucoside and tricin in methanolic extract of L. chinensis fruits, based on their mass spectra and fragmentation patterns. To the best of our knowledge, orientin, isoorientin, isovitexin and isorhamnetin-3-O-glucoside were found firstly in this plant. The free radical scavenging activity of the six antioxidants found is further examined by off-line DPPH assay. The results indicated that the free radical scavenging activity of orientin and isoorientin are stronger than those of two antioxidative drugs, vitamin C and baicalin. In addition, an HPLC-DAD method is firstly established for simultaneous determination of the six antioxidants in L. chinensis fruits. Tricin was found to be the major component in L. chinensis fruits.

  2. Validation of a RP-HPLC-DAD Method for Chamomile (Matricaria recutita Preparations and Assessment of the Marker, Apigenin-7-glucoside, Safety and Anti-Inflammatory Effect

    Directory of Open Access Journals (Sweden)

    Felipe Galeti Miguel

    2015-01-01

    Full Text Available Chamomile is a medicinal plant, which presents several biological effects, especially the anti-inflammatory effect. One of the compounds related to this effect is apigenin, a flavonoid that is mostly found in its glycosylated form, apigenin-7-glucoside (APG, in natural sources. However, the affectivity and safety of this glycoside have not been well explored for topical application. In this context, the aim of this work was to develop and validate a reversed-phase high-performance liquid chromatography (RP-HPLC-DAD method to quantify APG in chamomile preparations. Additionally, the safety and the anti-inflammatory potential of this flavonoid were verified. The RP-HPLC-DAD method was developed and validated with linearity at 24.0–36.0 μg/mL range (r=0.9994. Intra- and interday precision (RSD were 0.27–2.66% and accuracy was 98.27–101.21%. The validated method was applied in the analysis of chamomile flower heads, glycolic extract, and Kamillen cream, supporting the method application in the quality control of chamomile preparations. Furthermore, the APG safety was assessed by MTT cytotoxicity assay and mutagenic protocols and the anti-inflammatory activity was confirmed by a diminished TNF-α production showed by mice macrophages treated with APG following LPS treatment.

  3. Simultaneous extraction, identification and quantification of phenolic compounds in Eclipta prostrata using microwave-assisted extraction combined with HPLC-DAD-ESI-MS/MS.

    Science.gov (United States)

    Fang, Xinsheng; Wang, Jianhua; Hao, Jifu; Li, Xueke; Guo, Ning

    2015-12-01

    A simple and rapid method was developed using microwave-assisted extraction (MAE) combined with HPLC-DAD-ESI-MS/MS for the simultaneous extraction, identification, and quantification of phenolic compounds in Eclipta prostrata, a common herb and vegetable in China. The optimized parameters of MAE were: employing 50% ethanol as solvent, microwave power 400 W, temperature 70 °C, ratio of liquid/solid 30 mL/g and extraction time 2 min. Compared to conventional extraction methods, the optimized MAE can avoid the degradation of the phenolic compounds and simultaneously obtained the highest yields of all components faster with less consumption of solvent and energy. Six phenolic acids, six flavonoid glycosides and one coumarin were firstly identified. The phenolic compounds were quantified by HPLC-DAD with good linearity, precision, and accuracy. The extract obtained by MAE showed significant antioxidant activity. The proposed method provides a valuable and green analytical methodology for the investigation of phenolic components in natural plants.

  4. HPLC-DAD for the determination of three different classes of antifungals: method characterization, statistical approach, and application to a permeation study.

    Science.gov (United States)

    Miron, Diogo; Lange, Alini; Zimmer, Aline R; Mayorga, Paulo; Schapoval, Elfrides E S

    2014-12-01

    This study describes and characterizes methods for high-performance liquid chromatography diode array detection (HPLC-DAD) analysis of formulations containing molecules with antifungal activity of three different classes: terbinafine and butenafine (allylamines), miconazole and fluconazole (azoles), and geraniol, neral and geranial (monoterpenes). All methods used the same chromatographic column (RP18 ), enabling the analysis to be performed in a single batch. The specificity was extensively discussed through the establishment of purity peak methods. The analytical parameters (linearity, precision and accuracy) were calculated and discussed in detail using specific statistical approaches. All substances showed satisfactory results for chromatographic and analytical parameters. Limits of 1.3% to mean repeatability and 2.0% for intermediate precision are suggested as acceptance criteria in validation of methods by HPLC-DAD, in situations where there is no extensive pretreatment of the samples. The methods proved to be robust and significant factors were discussed regarding their influence on chromatographic parameters (retention time, resolution, tailing factor and column efficiency). Finally, the application of the developed methods was demonstrated by the results of a permeation study of the antifungal agents through bovine hoof membranes.

  5. Development and optimisation of an HPLC-DAD-ESI-Q-ToF method for the determination of phenolic acids and derivatives.

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    Annalaura Restivo

    Full Text Available A method for the HPLC-MS/MS analysis of phenols, including phenolic acids and naphtoquinones, using an amide-embedded phase column was developed and compared to the literature methods based on classical C18 stationary phase columns. RP-Amide is a recently developed polar embedded stationary phase, whose wetting properties mean that up to 100% water can be used as an eluent. The increased retention and selectivity for polar compounds and the possibility of working in 100% water conditions make this column particularly interesting for the HPLC analysis of phenolic acids and derivatives. In this study, the chromatographic separation was optimised on an HPLC-DAD, and was used to separate 13 standard phenolic acids and derivatives. The method was validated on an HPLC-ESI-Q-ToF. The acquisition was performed in negative polarity and MS/MS target mode. Ionisation conditions and acquisition parameters for the Q-ToF detector were investigated by working on collision energies and fragmentor potentials. The performance of the method was fully evaluated on standards. Moreover, several raw materials containing phenols were analysed: walnut, gall, wine, malbec grape, French oak, red henna and propolis. Our method allowed us to characterize the phenolic composition in a wide range of matrices and to highlight possible matrix effects.

  6. Validated Stability-Indicating HPLC-DAD Method for the Simultaneous Determination of Diclofenac Sodium and Diflunisal in Their Combined Dosage Form.

    Science.gov (United States)

    Shaalan, Rasha A; Belal, Tarek S

    2013-01-01

    A simple, rapid, and highly selective HPLC-DAD method was developed for the simultaneous determination of diclofenac sodium (DIC) and diflunisal (DIF) in pure form and in their combined formulation. Effective chromatographic separation was achieved using a Zorbax SB-C8 (4.6×250 mm, 5 μm particle size) column with a mobile phase composed of 0.05 M phosphoric acid, acetonitrile, and methanol in the ratio of 40:48:12 (by volume). The mobile phase was pumped isocratically at a flow rate of 1 mL/min, and quantification of the analytes was based on measuring their peak areas at 228 nm. The retention times for diflunisal and diclofenac were about 7.9 and 9.5 min, respectively. The reliability and analytical performance of the proposed HPLC procedure were statistically validated with respect to system suitability, linearity, ranges, precision, accuracy, specificity, robustness, detection, and quantification limits. Calibration curves were linear in the ranges of 5-100 μg/mL for both drugs with correlation coefficients >0.9998. The proposed method proved to be selective and stability-indicating by the resolution of the two analytes from four of their related substances and potential impurities as well as from forced-degradation (hydrolysis, oxidation, photolysis, and dry heat) products. The validated HPLC method was successfully applied to the analysis of DIC and DIF in their combined dosage form (suppositories). The proposed method made use of the diode array detector (DAD) as a tool for peak identity and purity confirmation.

  7. Determination of fourteen polyphenols in pulses by high performance liquid chromatography-diode array detection (HPLC-DAD) and correlation study with antioxidant activity and colour.

    Science.gov (United States)

    Giusti, Federica; Caprioli, Giovanni; Ricciutelli, Massimo; Vittori, Sauro; Sagratini, Gianni

    2017-04-15

    Pulses, which include lentils, beans, chickpeas, peas, and soybeans, provide an important source of proteins, dietary fibers, minerals and vitamins, as well as such important bioactive molecules as polyphenols. The presence of polyphenols is often related to the colour of the pulse and to its antioxidant activity. The aim of this work was to set up a new HPLC-DAD method for simultaneously analysing 14 polyphenolic compounds, including two anthocyanins, in different varieties of pulses and to correlate the polyphenol content with the seed coat colour and the antioxidant activity. The total content of the analysed polyphenols ranged from 3mg/kg for dehulled red lentils to 1630.5mg/kg for ruviotto beans. Samples with dark testa (or seed coat), namely black lentils and diavoli beans, had higher antioxidant activity than those with pale testa, and a positive correlation was found between total phenolic content (TPC) and IC50 for dark coloured varieties.

  8. A Stability-Indicating HPLC-DAD Method for Determination of Ferulic Acid into Microparticles: Development, Validation, Forced Degradation, and Encapsulation Efficiency

    OpenAIRE

    Jessica Mendes Nadal; Maria da Graça Toledo; Yasmine Mendes Pupo; Josiane Padilha de Paula; Paulo Vitor Farago; Sandra Maria Warumby Zanin

    2015-01-01

    A simple stability-indicating HPLC-DAD method was validated for the determination of ferulic acid (FA) in polymeric microparticles. Chromatographic conditions consisted of a RP C18 column (250 mm × 4.60 mm, 5 μm, 110 Å) using a mixture of methanol and water pH 3.0 (48 : 52 v/v) as mobile phase at a flow rate of 1.0 mL/min with UV detection at 320 nm. The developed method was validated as per ICH guidelines with respect to specificity, linearity, limit of quantification, limit of detection, ac...

  9. Identification and characterization of low molecular weight polyphenols in berry leaf extracts by HPLC-DAD and LC-ESI/MS.

    Science.gov (United States)

    Oszmiański, Jan; Wojdyło, Aneta; Gorzelany, Józef; Kapusta, Ireneusz

    2011-12-28

    This paper reports the results of qualitative and quantitative analyses of low molecular weight phenolics from five berry leaf extract using HPLC-DAD and LC-ESI/MS. The identification of the black currant, raspberry, bilberry, honeysuckle, and strawberry leaf phytochemicals was based on the comparison of UV-vis absorption maxima (λ(max)) and mass spectral analysis. The peak identification in samples was also based on comparisons of the retention times (t(R)) of the isolated phytochemical standards. Knowledge of the precise phenolic profile of berry leaves may offer a scientific basis to put the underutilized berry leaves to good use as very cheap raw materials for polyphenol extract production. These studies indicated that these leaves can be used as a good and cheap source of bioactive constituents. These results suggest that berry leaves are a potential source of phenolics and have potential pro-healthy properties to contribute to human health.

  10. Rapid and accurate hplc-dad method for the determination of the herbicide bispyribac-sodium in surface water, and its validation

    Directory of Open Access Journals (Sweden)

    Márcia H. S. Kurz

    2009-01-01

    Full Text Available A new method is described for the determination of the herbicide bispyribac-sodium in surface water, especially from river and irrigated rice water samples. The method involves extraction in solid phase and quantification by high performance liquid chromatography with diode array detection (HPLC-DAD. After optimization of the extraction and separation parameters, the method was validated. The method presented average recoveries of 101.3 and 97.7%, under repeatability and intermediate precision conditions, respectively, with adequate precision (RSD from 0.9 to 7.5%. The method was applied for the determination of bispyribac-sodium in surface water samples with a limit of detection of 0.1 μg L-1.

  11. Analysis of Naturally Occurring Phenolic Compounds in Aromatic Plants by RP-HPLC Coupled to Diode Array Detector (DAD and GC-MS after Silylation

    Directory of Open Access Journals (Sweden)

    Charalampos Proestos

    2013-03-01

    Full Text Available The following aromatic plants of Greek origin, Origanum dictamnus (dictamus, Eucalyptus globulus (eucalyptus, Origanum vulgare L. (oregano, Mellisa officinalis L. (balm mint and Sideritis cretica (mountain tea, were examined for the content of phenolic substances. Reversed phase HPLC coupled to diode array detector (DAD was used for the analysis of the plant extracts. The gas chromatography-mass spectrometry method (GC-MS was also used for identification of phenolic compounds after silylation. The most abundant phenolic acids were: gallic acid (1.5–2.6 mg/100 g dry sample, ferulic acid (0.34–6.9 mg/100 g dry sample and caffeic acid (1.0–13.8 mg/100 g dry sample. (+-Catechin and (−-epicatechin were the main flavonoids identified in oregano and mountain tea. Quercetin was detected only in eucalyptus and mountain tea.

  12. Comprehensive identification of 125 multifarious constituents in Shuang-huang-lian powder injection by HPLC-DAD-ESI-IT-TOF-MS.

    Science.gov (United States)

    Sun, Hongyang; Liu, Meixian; Lin, Zongtao; Jiang, Haixiu; Niu, Yanyan; Wang, Hong; Chen, Shizhong

    2015-11-10

    A high-performance liquid chromatography-diode array detector-electrospray ionization-ion trap-time of flight-mass spectrometry (HPLC-DAD-ESI-IT-TOF-MS) method was established for excellent separation and structural identification of constituents in Shuang-huang-lian powder injection (SHLPI). The typical ultraviolet absorptions, accurate empirical molecular formula and reasonable fragmentation mechanisms of these ingredients were used for their structural elucidation. In consequence, 125 constituents (33 phenolic acids, 29 flavonoids, 32 phenylethanoid glycosides, 15 iridoid glycosides, 8 lignans, 3 amino acids and 2 purines nucleosides, 2 quinoid glycosides and 1 alkylbenzene glycoside) were either unequivocally identified or tentatively characterized by comparing authentic standards or published data. The result showed that this study could provide valuable information for the quality control and further investigation of SHLPI formula.

  13. Simultaneous analysis of anthocyanin and non-anthocyanin flavonoid in various tissues of different lotus (Nelumbo cultivars by HPLC-DAD-ESI-MS(n.

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    Sha Chen

    Full Text Available A validated HPLC-DAD-ESI-MS(n method for the analysis of non-anthocyanin flavonoids was applied to nine different tissues of twelve lotus genotypes of Nelumbo nucifera and N. lutea, together with an optimized anthocyanin extraction and separation protocol for lotus petals. A total of five anthocyanins and twenty non-anthocyanin flavonoids was identified and quantified. Flavonoid contents and compositions varied with cultivar and tissue and were used as a basis to divide tissues into three groups characterized by kaempferol and quercetin derivatives. Influences on flower petal coloration were investigated by principal components analyses. High contents of kaempferol glycosides were detected in the petals of N. nucifera while high quercetin glycoside concentrations occurred in N. lutea. Based on these results, biosynthetic pathways leading to specific compounds in lotus tissues are deduced through metabolomic analysis of different genotypes and tissues and correlations among flavonoid compounds.

  14. A Novel Method HPLC-DAD Analysis of the Contentsof Moutan Cortexand Paeoniae Radix Alba with Similar Constituents-Monoterpene Glycosides in Guizhi Fuling Wan

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    Shuyun Wang

    2014-11-01

    Full Text Available A variety of traditional Chinese medical formulations contain two or more herbs from the same genus or family. Although these herbs may have a similar appearance and constituents, they usually have different pharmacodynamic actions. A series of qualitative and quantitative analysis methods are developed to determine one or more compounds for quality control of medicine. As far as we know, no method has been found to determine the real ratio of the two herbs along with the prescription. In this study, we used HPLC-DAD as a way to determine the content of Moutan cortex (M and Paeoniae radix alba (P in GuizhiFuling Wan (GZFLW. An effective, accurate and reliable HPLC-DAD method was developed for detecting the content of M and P in GZFLW through the analysis of four monoterpeneglycosides, namely, galloylpaeoniflorin (1, paeoniflorin (2, mudanpioside C (3 and benzoylpaeoniflorin (4. Due to the different UV characteristics of the compounds, the detection wavelength was 270 nm for 1 and 2, while 3 and 4 were monitored at 254 nm and 230 nm, respectively. Four equations were put forward to describe the relationship between content of M as well as P and the four monoterpene glycosides in GZFLW. After validation, all the accuracies of the M and P contents in GZFLW were within 10%. The result showed that the method could be successfully applied to analyze the contents of M and P in GZFLW. Moreover, our method may be more widely used to control the quality of proprietary Chinese medicines, especially for those containing the same genus or family herbs, in industrial GMP production.

  15. Combination of hollow fiber liquid phase microextraction followed by HPLC-DAD and multivariate curve resolution to determine antibacterial residues in foods of animal origin.

    Science.gov (United States)

    Tajabadi, Fateme; Ghambarian, Mahnaz; Yamini, Yadollah; Yazdanfar, Najmeh

    2016-11-01

    In the present research, a carrier mediated hollow fiber based liquid-phase microextraction approach (HF-LPME) prior to high performance liquid chromatography-diode array detection (HPLC-DAD) was developed for the simultaneous determination of the antibacterial residues of four tetracyclines (TCs) and five quinolones (QNs), which are commonly used as veterinary medicines. In order to obtain high extraction efficiency, the parameters affecting HF-LPME were optimized using a three-factor and three-level Box-Behnken design under response surface methodology. This method was validated according to the recommendations of the Food and Drug Administration (FDA), and, for the first time, successfully applied to a wide range of animal source food samples such as fish, milk, and honey as well as the liver and muscles of lamb and chicken. Analytical performance was determined in terms of linearity, intra- and inter-assay precision, detection and quantification limits, matrix effect, accuracy, and drug stability in real samples. Detection and quantitation limits for the different antibiotics ranged between 0.5-20ngg(-1) and 1.25-40ngg(-1), respectively. Intra and inter-day repeatability, expressed as the relative standard deviation, were in the ranges of 3.4-10.7% and 5.0-11.5%, respectively. The procedure allows good preconcentration factors of 175-700. The results of the validation process proved that the method is suitable for determining TCs and QNs residues in surveillance programs. Finally, the applicability of the proposed method was successfully confirmed by the extraction and determination of nine antibiotics in various animal source food samples. The importance of this methodology relies on the combination of HF-LPME/HPLC-DAD second-order data with multivariate curve resolution-alternative least squares (MCR-ALS) algorithm, which improves the resolution of some overlapped chromatograms and, hence, increases the accuracy and repeatability of drug determination.

  16. METODOLOGÍA PARA LA DETERMINACIÓN DE RESIDUOS DE FUNGICIDAS BENZIMIDAZÓLICOS EN FRESA Y LECHUGA POR HPLC-DAD.

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    Jose Jairo Dangond Araujo

    2008-03-01

    Full Text Available Los benzimidazoles son fungicidas de acción sistémica muy utilizados en la protección de cultivos de frutas y hortalizas. En este trabajo se validó una metodología para la determinación de residuos de benomyl, carbendazim y tiabendazol, en fresa y lechuga por cromatografía líquida de alta eficiencia con detector de arreglo de diodos (HPLC-DAD. Los residuos de benomyl se determinaron luego de su conversión a carbendazim. La extracción de los residuos de las muestras se realizó con acetato de etilo y la limpieza se llevó a cabo por cromatografía de permeación en gel (GPC. La determinación analítica se realizó por HPLC-DAD en fase reversa. La metodología es selectiva, específica, precisa y exacta. Las curvas de calibración son lineales en un rango de concentración de 1,24 a 6,19 mg/kg con límites de detección de 0,27 y 0,40 mg/kg y límites de cuantificación de 0,85 y 1,35 mg/kg para carbendazim y tiabendazol respectivamente. Los porcentajes de recuperación son del orden del 90%. No se encontraron residuos de estos compuestos en muestras recolectadas en algunos municipios de Cundinamarca, Colombia.

  17. Simultaneous Determination of Eight Phenolic Acids, Five Saponins and Four Tanshinones for Quality Control of Compound Preparations Containing Danshen-Sanqi Herb-pair by HPLC-DAD.

    Science.gov (United States)

    Yao, Hong; Huang, Xiaomei; Li, Shaoguang; Wu, Youjia; Lin, Xinhua; Shi, Peiying

    2017-01-01

    successfully applied to the quality control of five compound preparations containing DS-SQ herb-pair.Additionally, it found that the favorable dosage forms for prescriptions containing DS-SQ herb-pair could be solid preparations. Abbreviations used: DS: Salviae miltiorrhizae; SQ: Panaxnotoginseng; HPLC: high-performance liquid chromatography; DAD: diode array detector; LOD: limit of detection; LOQ: limit of quantification; TCMs: Traditional Chinese medicines; GDDP: Guanxin Danshen dripping pills; FDDP: Fufang Danshen dripping pills; FDT: Fufang Danshen tablets; FDC: Fufang Danshen capsules; GP: Guanxin pills Key Messages: The HPLC-DAD analysis successfully fulfilled the simultaneous determination of 17 compounds (including three types of authentic bioactive components, 8 phenolic acids, 4 tanshinones, and 5 saponins) in DS-SQ herb-pair within 70 min with the routine HPLC for the first time. The results also demonstrated that solid preparations could be the favorable dosage forms for those prescriptions containing DS-SQ herb-pair due to the instability of saponins from SQ, when the components of DS and SQ were coexisting in solution. The study provides a promising tool for quality control of the preparations containing the DS-SQ herb-pair.

  18. Simultaneous Determination of Eight Phenolic Acids, Five Saponins and Four Tanshinones for Quality Control of Compound Preparations Containing Danshen-Sanqi Herb-pair by HPLC-DAD

    Science.gov (United States)

    Yao, Hong; Huang, Xiaomei; Li, Shaoguang; Wu, Youjia; Lin, Xinhua; Shi, Peiying

    2017-01-01

    time by routine HPLC.The presented method was successfully applied to the quality control of five compound preparations containing DS-SQ herb-pair.Additionally, it found that the favorable dosage forms for prescriptions containing DS-SQ herb-pair could be solid preparations. Abbreviations used: DS: Salviae miltiorrhizae; SQ: Panaxnotoginseng; HPLC: high-performance liquid chromatography; DAD: diode array detector; LOD: limit of detection; LOQ: limit of quantification; TCMs: Traditional Chinese medicines; GDDP: Guanxin Danshen dripping pills; FDDP: Fufang Danshen dripping pills; FDT: Fufang Danshen tablets; FDC: Fufang Danshen capsules; GP: Guanxin pills Key Messages: The HPLC-DAD analysis successfully fulfilled the simultaneous determination of 17 compounds (including three types of authentic bioactive components, 8 phenolic acids, 4 tanshinones, and 5 saponins) in DS-SQ herb-pair within 70 min with the routine HPLC for the first time. The results also demonstrated that solid preparations could be the favorable dosage forms for those prescriptions containing DS-SQ herb-pair due to the instability of saponins from SQ, when the components of DS and SQ were coexisting in solution. The study provides a promising tool for quality control of the preparations containing the DS-SQ herb-pair. PMID:28216885

  19. Development and validation of HPLC-DAD-CAD-MS(3) method for qualitative and quantitative standardization of polyphenols in Agrimoniae eupatoriae herba (Ph. Eur).

    Science.gov (United States)

    Granica, Sebastian; Krupa, Katarzyna; Kłębowska, Agnieszka; Kiss, Anna K

    2013-12-01

    A reversed phase high performance liquid chromatography method (HPLC) coupled with a diode array, charged aerosol detector or mass spectrometer was developed for the quantitative and qualitative standardization of Agrimoniae eupatoriae herba (Ph. Eur). Twenty four constituents comprising phenolic acids, flavan-3-ol derivatives, ellagitannin and flavonoids were fully or partially identified. Eight of detected compounds were reported from common agrimony for the first time. Fourteen major polyphenols were quantified using validated HPLC method with UV-vis and corona charged detection. Both detectors were shown to be equal for the quantification of selected polyphenols. Some limitations of universal response of corona charged detector for phenolic compounds were discussed. Using obtained data for DAD detector the sum of tannins, flavonoids and phenolic acids quantified in examined plant material was determined. Investigated samples contained 8.2-10.9mg/g of flavonoids, 6.3-10.9mg/g of tannins (among which agrimoniin was dominating constituent 2.6-5.4mg/g) and 0.6-0.9mg/g of phenolic acids proving that flavonoids should be considered as second major group of constituents in common agrimony.

  20. Simultaneous Quantification of Limonin, Two Indolequinazoline Alkaloids, and Four Quinolone Alkaloids in Evodia rutaecarpa (Juss. Benth by HPLC-DAD Method

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    Pei-ting Zhang

    2013-01-01

    Full Text Available A simple and efficient HPLC-DAD (225 nm method was developed and validated for the simultaneous determination of limonin and six key alkaloids (evodiamine, rutaecarpine, 1-methyl-2-undecyl-4(1H-quinolone, evocarpine, 1-methy-2-[(6Z,9Z]-6,9-pentadecadienyl-4-(1H-quinolone, and dihydroevocarpine in Evodia rutaecarpa (Juss. Benth, which has been widely used as one of the Traditional Chinese Medicines. The chromatographic separation was carried out on a Hypersil BDS C18 column, and gradient elution was employed with a mobile phase containing acetonitrile and water. Contents of the analytes in 18 batches of samples were analyzed by ultrasonic extraction with ethanol and water mixture (80 : 20, v/v followed by HPLC analysis. Separation of the seven analytes was achieved within 60 min with good linearity (. The RSD of both the intraday and interday precision was below 1.85%. The accuracy at different concentrations was within the range of 97.91 to 100.49%. Hierarchical clustering analysis was performed to differentiate and classify the samples based on the contents of the seven constituents. This study indicated that the quality control of E. rutaecarpa could be simplified to the measurement of four constituents, and that limonin, 1-methyl-2-undecyl-4(1H-quinolone, and dihydroevocarpine should also be served as the chemical markers together with evodiamine for the quality control of Evodia rutaecarpa (Juss. Benth.

  1. Development and validation of HPLC-DAD method for the simultaneous determination of amoxicillin, metronidazole and rabeprazole sodium. Application to spiked simulated intestinal fluid samples.

    Science.gov (United States)

    Sabry, S M; Abdel-Hay, M H; Belal, T S; Mahgoub, A A

    2015-09-01

    This work deals with the development, validation and application of an HPLC-DAD method for the determination of a ternary mixture containing amoxicillin (AX), metronidazole (MZ) and the proton pump inhibitor rabeprazole sodium (RB). This triple therapy is used for treatment of Helicobacter pylori infection. Effective chromatographic separation between the three drugs was achieved using Thermo Hypersil BDS-C8 (4.6×250mm, 5μm particle size) column and a mobile phase composed of phosphate buffer pH 7 and acetonitrile (70: 30, by volume). The mobile phase was pumped isocratically at a flow rate of 1 mL/min. Quantification of the analytes was based on measuring their peak areas at 230nm for both AX and RB, and at 319nm for MZ. AX, MZ and RB eluted at retention times 2.36, 3.55 and 8.72min respectively. The reliability and analytical performance of the proposed HPLC procedure were statistically validated with respect to linearity, ranges, precision, accuracy, selectivity, robustness, detection and quantification limits. The linear dynamic ranges were 25-250, 25-250 and 5-50μg/mL for AX, MZ and RB respectively with correlation coefficients>0.9998. The validated method was successfully applied to the analysis of several laboratory-prepared mixtures as well as simulated intestinal fluid samples spiked with the three drugs.

  2. HPLC-DAD-MS/MS profiling of phenolics from Securigera securidaca flowers and its anti-hyperglycemic and anti-hyperlipidemic activities

    Directory of Open Access Journals (Sweden)

    Rana M. Ibrahim

    2015-04-01

    Full Text Available Abstract Securigera securidaca (L. Degen & Döefl., Fabaceae, has been widely used in the Iranian, Indian and Egyptian folk medicine as antidiabetic and anti-hyperlipidemic remedy. Phenolic profiling of the ethanolic extract (90% of the flowers of S. securidaca was performed via HPLC-DAD-MS/MS analysis in the positive and negative ion modes. The total polyphenols and flavonoids in the flowers were determined colorimetrically, and the quantification of their components was carried out using HPLC-UV. Total phenolics and flavonoids estimated as gallic acid and rutin equivalents were 82.39 ± 2.79 mg/g and 48.82 ± 1.95 mg/g of the dried powdered flowers, respectively. HPLC-DAD-MS/MS analysis of the extract allowed the identification of 39 flavonoids and eight phenolic acids. Quantitative analysis of some flavonoids and phenolics (mg/100 g powdered flowers revealed the presence of isoquercetrin (3340 ± 2.1, hesperidin (32.09 ± 2.28, naringin (197.3 ± 30.16, luteolin (10.247 ± 0.594, chlorogenic acid (84.22 ± 2.08, catechin (3.94 ± 0.57 and protocatechuic acid (34.4 ± 0.15, in the extract. Moreover, the acute toxicity, hypoglycemic and hypolipidemic effects of the extract were investigated using alloxan induced diabetes in rats in a dose of 100, 200, and 400 mg/kg bwt. The ethanolic extract was safe up to a dose of 2000 mg/kg. All tested doses of the flower extract showed marked decrease in blood glucose level by 31.78%, 66.41% and 63.8% at 100, 200 and 400 mg/kg bwt, respectively, at p < 0.05. Regarding the anti-hyperlipidemic effect, a dose of 400 mg/kg of the flower extract showed the highest reduction in serum triacylglycerides and total cholesterol levels (68.46% and 51.50%, respectively at p < 0.05. The current study proved the folk use of the flowers of S. securidaca as anti-diabetic and anti-hyperlipidemic agent which could be attributed to its high phenolic content.

  3. Simultaneous Determination of 14 Phenolic Compounds in Grape Canes by HPLC-DAD-UV Using Wavelength Switching Detection

    Directory of Open Access Journals (Sweden)

    Ang Zhang

    2013-11-01

    Full Text Available The paper described a novel chromatographic method for the simultaneous determination of phenolic compounds such as gallic, protocatechuic, vanillic, caffeic, syringic, p-coumaric and salicylic acid, (+-catechin, (‒-epicatechin, rutin, morin, quercetin, coumarin and trans-resveratrol at their maximum absorbance wavelengths (MAW employing reverse-phase high performance liquid chromatography combined with DAD and UV detection via detection wavelength switching. The method was based on MAW acquisition by DAD and quantification by UV. The separation process was performed on a Shim-Pack VP-ODS C18 column (250 mm × 4.6 mm, 5 μm held at 30 °C, utilizing 3.0% acetic acid and acetonitrile as mobile phase at a flow rate of 0.8 mL/min in the gradient elution mode. The method was fully validated in terms of linearity (r2 > 0.9990, 10‒350 mg/L, precision (both intra-day and inter-day RSD < 4.22%, accuracy (97.31%‒104.66%, specificity, robustness (0.59% < RSD < 2.86%, limit of detection and quantification. The switching method significantly improved the sensitivities of most phenolics studied in comparison with the standard constant wavelength detection (280 nm. The proposed method has been successfully applied to the determination of 14 phenolic compounds in 89 varieties of one-year-old Chinese grape one-year-canes. Grape canes contain many phenolics, especially trans-resveratrol, (‒-epicatechin, and (+-catechin.

  4. Development and validation of an HPLC-DAD method for simultaneous determination of cocaine, benzoic acid, benzoylecgonine and the main adulterants found in products based on cocaine.

    Science.gov (United States)

    Floriani, Gisele; Gasparetto, João Cleverson; Pontarolo, Roberto; Gonçalves, Alan Guilherme

    2014-02-01

    Here, an HPLC-DAD method was developed and validated for simultaneous determination of cocaine, two cocaine degradation products (benzoylecgonine and benzoic acid), and the main adulterants found in products based on cocaine (caffeine, lidocaine, phenacetin, benzocaine and diltiazem). The new method was developed and validated using an XBridge C18 4.6mm×250mm, 5μm particle size column maintained at 60°C. The mobile phase consisted of a gradient of acetonitrile and ammonium formate 0.05M - pH 3.1, eluted at 1.0mL/min. The volume of injection was 10μL and the DAD detector was set at 274nm. Method validation assays demonstrated suitable sensitivity, selectivity, linearity, precision and accuracy. For selectivity assay, a MS detection system could be directly adapted to the method without the need of any change in the chromatographic conditions. The robustness study indicated that the flow rate, temperature and pH of the mobile phase are critical parameters and should not be changed considering the conditions herein determined. The new method was then successfully applied for determining cocaine, benzoylecgonine, benzoic acid, caffeine, lidocaine, phenacetin, benzocaine and diltiazem in 115 samples, seized in Brazil (2007-2012), which consisted of cocaine paste, cocaine base and salt cocaine samples. This study revealed cocaine contents that ranged from undetectable to 97.2%, with 97 samples presenting at least one of the degradation products or adulterants here evaluated. All of the studied degradation products and adulterants were observed among the seized samples, justifying the application of the method, which can be used as a screening and quantification tool in forensic analysis.

  5. Quantity and quality of black carbon molecular markers as obtained by two chromatographic methods (GC-FID and HPLC-DAD) - How do results compare?

    Science.gov (United States)

    Schneider, M. P. W.; Smittenberg, R. H.; Dittmar, T.; Schmidt, M. W. I.

    2009-04-01

    Chars produced by wildfires are an important source of black carbon (BC) in the environment. After their deposition on the soil surface they can be distributed into rivers, marine waters and sediments. The analysis of benzenepolycarboxylic acids (BPCAs) as a quantitative measure for black carbon (BC) in soil and sediment samples is a well-established method (Glaser et al., 1998; Brodowski et al., 2005). Briefly, the nitric acid oxidation of fused aromatic ring systems in BC forms eight molecular markers (BPCAs), which can be assigned to BC, and which subsequently can be quantified by GC-FID (gas chromatography with flame ionization detector). Recently, this method was modified for the quantification of BC in seawater samples using HPLC-DAD (High performance liquid chromatography with diode array detector) for the determination of individual BPCAs (Dittmar, 2008). A direct comparison of both analytical techniques is lacking but would be important for future data comparison aimed at the calculation of global BC budgets. Here we present a systematic comparison of the two BPCA quantification methods. We prepared chars under well-defined laboratory conditions. In order to cover a broad spectrum of char properties we used two sources of biomass and a wide range of pyrolysis temperatures. Chestnut hardwood chips (Castanea sativa) and rice straw (Oryza sativa) were pyrolysed at temperatures between 200 and 1000°C under a constant N2 stream. The maximum temperatures were held constant for 5 hours (Hammes et al., 2006). The BC contents of the chars have been analysed using the BPCA extraction method followed by either GC-FID or HPLC-DAD quantification. Preliminary results suggest that both methods yield similar total quantities of BPCA, and also the proportions of the individual markers are similar. Ongoing experiments will allow for a more detailed comparison of the two methods. The BPCA composition of chars formed at different temperatures and from different precursor

  6. Nonanthocyanin secondary metabolites of black raspberry (Rubus occidentalis L.) fruits: identification by HPLC-DAD, NMR, HPLC-ESI-MS, and ESI-MS/MS analyses.

    Science.gov (United States)

    Paudel, Liladhar; Wyzgoski, Faith J; Scheerens, Joseph C; Chanon, Ann M; Reese, R Neil; Smiljanic, Danijela; Wesdemiotis, Chrys; Blakeslee, Joshua J; Riedl, Kenneth M; Rinaldi, Peter L

    2013-12-11

    Nonanthocyanin secondary metabolites potentially contributing to the antiproliferative bioactivity of black raspberry ( Rubus occidentalis L.) fruits were extracted in ethyl acetate and isolated by semipreparative and analytical HPLC and analyzed by NMR, HPLC-ESI-MS, and ESI-MS/MS techniques. Here we present complete and partial structures of a variety of the chemical entities such as quercetin 3-glucoside, quercetin 3-rutinoside, myricetin glucoside, dihydrokaempferol glucoside, benzoic acid β-d-glucopyranosyl ester, 3,4-dihydroxybenzoic acid, epicatechin, caffeic acid, p-coumaric acid, p-coumaryl glucoside, p-coumaryl sugar ester, ellagic acid, methyl ellagic acid acetylpentose, methyl ellagic acid valerylpentose, trans-piceid, phloretin glucoside (phloridzin), dihydrosinapic acid, salicylic acid β-d-glucopyranosyl ester, a salicylic acid derivative without attached sugar, p-alkylphenyl glucoside, and a citric acid derivative. To our knowledge, 15 of these compounds were not previously reported in black raspberry fruits.

  7. 冠心宁注射液中主成分在线HPLC-DAD-CL抗氧化活性测定%On-line antioxidant activity determination of main ingredients in Guan-Xin-Ning injection by HPLC-DAD-CL

    Institute of Scientific and Technical Information of China (English)

    李意; 阮鸣; 罗建光; 孔令义

    2012-01-01

    采用HPLC-DAD-CL方法同时分析了冠心宁注射液中15个成分的抗氧化活性.冠心宁注射液是由丹参和川芎组方的传统中药制剂,主要用于治疗冠心病和心绞痛.该分析方法的条件被优化、验证,并用于14批冠心宁注射液中15个酚酸成分的H2O2抑制率测定,结果显示15个化合物的抗氧化活性具有明显的差异,且丹参素、原儿茶醛和迷迭香酸是冠心宁注射液整体抗氧化活性的主要贡献成分(25.82%-51.19%).此外,通过PCA方法可将14批冠心宁注射液分成5组,此5组与5个生产厂家对应.因此,本论文提出的HPLC-DAD-CL方法简单、快速、成本低,可用于复杂体系的抗氧化成分的筛选、揭示冠心宁注射液的抗氧化机制,亦可从活性角度用于其质量控制.%A novel on-line method using high performance liquid chromatography (HPLC) with a diode array detector (DAD) and a chemiluminescence (CL) detector was successfully developed for simultaneously analyzing the antioxidant efficacies of 15 components in Guan-Xin-Ning (GXN) injection,a traditional Chinese medicinal preparation consisting of Radix Salvia miltiorrhiza and Rhizoma Ligusticum chuanxiong for the treatment of coronary heart disease and angina pectoris.The analytical conditions were optimized,validated and applied to determine H2O2 inhibition rates of the 15 phenolic acids in GXN injections from 14 different production batches.The results showed that antioxidant diversities were found in the 15 compounds and that danshensu,protocatechuic aldehyde and rosmarinic acid were largely responsible for the antioxidant activity of GXN injection (25.82%-51.19%).In addition,14 batches of GXN injections were divided into five groups corresponding with five different factories by principal component analysis (PCA) assay.Therefore,the proposed HPLC-DAD-CL method was simple,rapid and low-cost for screening antioxidant constituents in a complex system,which can be used to reveal

  8. Toxicological screening of human plasma by on-line SPE-HPLC-DAD: identification and quantification of acidic and neutral drugs.

    Science.gov (United States)

    Mut, Ludmila; Grobosch, Thomas; Binscheck-Domaß, Torsten; Frenzel, Wolfgang

    2016-03-01

    A multi-analyte screening method for the quantification of 50 acidic/neutral drugs in human plasma based on on-line solid-phase extraction (SPE)-HPLC with photodiode array detection (DAD) was developed, validated and applied for clinical investigation. Acetone and methanol for protein precipitation, three different SPE materials (two electro-neutral, one strong anion-exchange, one weak cation-exchange) for on-line extraction, five HPLC-columns [one C18 (GeminiNX), two phenyl-hexyl (Gemini C6 -Phenyl, Kinetex Phenyl-Hexyl) and two pentafluorophenyl (LunaPFP(2), KinetexPFP)] for analytical separation were tested. For sample pre-treatment, acetone in the ratio 1:2 (plasma:acetone) showed a better baseline and fewer matrix peaks in the chromatogram than methanol. Only the strong anion-exchanger SPE cartridge (StrataX-A, pH 6) allowed the extraction of salicylic acid. Analytical separation was carried out on a Gemini C6 -Phenyl column (150 × 4.6 mm, 3 µm) using gradient elution with acetonitrile-water 90:10 (v/v) and phosphate buffer (pH 2.3). Linear calibration curves with correlation coefficients r ≥ 0.9950/0.9910 were obtained for 46/four analytes. Additionally, this method allows the quantification of 23 analytes for therapeutic drug monitoring. Limits of quantitation ranged from 0.1 (amobarbital) to 23 mg/L (salicylic acid). Inter-/intra-day precisions of quality control samples (low/high) were better than 13% and accuracy (bias) ranged from -14 to 10%. A computer-assisted database was created for automated detection of 223 analytes of toxicological interests. Four cases of multi-drug intoxications are presented.

  9. Simultaneous determination and assignment of 13 major flavonoids and glycyrrhizic acid in licorices by HPLC-DAD and Orbirap mass spectrometry analyses.

    Science.gov (United States)

    Wei, Shan-Shan; Yang, Min; Chen, Xin; Wang, Qiu-Rong; Cui, Ya-Jun

    2015-03-01

    To determine 13 flavonoids and glycyrrhizic acid in licorice (Glycyrrhiza spp.), several samples from different areas were examined by HPLC-DAD analysis. The analysis was performed on a Zorbax Extend-C18 (250 mm × 4.6 mm, 5 μm) column connected with a Zorbax Extend guard column (20 mm × 4.6 mm, 5 μm). The mobile phase consisted of (A) acetonitrile and (B) 0.026% aqueous H3PO4 (VV) using a gradient elution of 20%-25% A at 0-20 min, 25%-33% A at 20-30 min, 33%-50% A at 30-55 min, 50%-60% A at 55-65 min, and 60% A between 65 min and 80 min, and peaks were detected at 280 nm. The fourteen compounds were assigned by HPLC-Orbitrap MS methods. The regression coefficient for the linear equations for the 14 compounds ranged between 0.9998 and 1. The limits of detection and quantification lay in the range of 0.032-2.461 and 0.154-8.202 μg·mL(1), respectively. The relative recovery rates for the 14 compounds were in the range of 93.90%-106.73% with RSDs being less than 5%. Coefficient variations for intra-day and inter-day precisions were in the range of 0.27%-2.38% and 0.31%-3.51%, respectively. In summary, the validated method was applied to the simultaneous determination of the 14 components in 29 different licorice samples and was proven to be suitable for quality evaluation of licorices and their active fractions.

  10. Estimation of psoralen from herbal formulations containing Psoralea corylifolia using the RP-HPLC-DAD method and its application to a pharmacokinetic study

    Directory of Open Access Journals (Sweden)

    Sunita Shailajan

    2012-01-01

    Full Text Available Background: Psoralea corylifolia L. (Fabaceae, seeds is used in many formulations for the treatment of a wide range of ailments. Aims and Objective: To develop a reverse phase high-performance liquid chromatography-photodiode array detector (RP-HPLC-DAD method for quantitation of psoralen from P. corylifolia and its related formulations. Materials and Methods: Separation and detection of psoralen from various herbal formulations was achieved on reversed phase Cosmosil C 18 column using acetonitrile: Distilled water (40: 60, v/v; flow rate - 1.0 mL/minute and the PDA detector (247 nm. The method was validated as per the norms of the International Conference on Harmonisation (ICH guidelines and applied to study the pharmacokinetics of an oil-based Ayurvedic preparation, in terms of bioavailable psoralen. Results: The HPLC method showed a linear detector response from 20.0 to 5000.0 ng/mL (r 2 =0.9998 for psoralen. The content of psoralen in P. corylifolia and its marketed formulations was determined, which showed remarkable variations as per the nature and complexity of the formulation. The absorption and elimination profile of psoralen, from an oil-based Ayurvedic preparation, was developed on its topical application in rabbits. Psoralen was detected in the plasma 0.25 hours post application of Bakuchi Taila with 0.46% bioavailability. Conclusion: The method was found to be sensitive, accurate and reproducible. Therefore, it can be recommended for marker-based standardisation and quality assurance of P. corylifolia and its formulations. It can also be applied to study the pharmacokinetic profile of the traditional preparations of P. corylifolia.

  11. Metal organic frameworks (MOFs) for magnetic solid-phase extraction of pyrazole/pyrrole pesticides in environmental water samples followed by HPLC-DAD determination.

    Science.gov (United States)

    Ma, Jiping; Yao, Zhidan; Hou, Liwei; Lu, Wenhui; Yang, Qipeng; Li, Jinhua; Chen, Lingxin

    2016-12-01

    Magnetic metal-organic frameworks (MOFs, [MIL-101]) were prepared and used as magnetic solid-phase extraction (MSPE) adsorbents for preconcentration of four kinds of pyrazole/pyrrole pesticides (flusilazole, fipronil, chlorfenapyr, and fenpyroximate) in environmental water samples, followed by high-performance liquid chromatography-diode-array detector (HPLC-DAD) determination. Several variables affecting MSPE efficiency were systematically investigated, including amount of MIL-101, extraction time, sample pH, salt concentration, type of desorption solvent and desorption number of times. Under optimized conditions, excellent linearity was achieved in the range of 5.0-200.0μg/L for flusilazole and fipronil, and 2.0-200.0μg/L for chlorfenapyr and fenpyroximate, with correlation coefficients r>0.9911. Limits of detection and quantification were 0.3-1.5μg/L and 1.0-5.0μg/L, respectively. The intra-day and inter-day precision (relative standard deviation, n=6, %) at three spiked levels were 1.1-5.4% and 3.9-7.8% in terms of peak area, respectively. The method recoveries at three fortified concentration levels ranged from 81.8% to 107.5% for reservoir water samples, 81.0-99.5% for river water samples, and 80.2-106.5% for seawater samples. The developed MOFs based MSPE coupled with HPLC method proved to be a convenient, rapid and eco-friendly alternative to the sensitive determination of pyrazole/pyrrole pesticides with high repeatability and excellent practical applicability.

  12. Investigating the stability of the nonionic surfactants tocopheryl polyethylene glycol succinate and sucrose laurate by HPLC-MS, DAD, and CAD.

    Science.gov (United States)

    Christiansen, Anne; Backensfeld, Thomas; Kühn, Silke; Weitschies, Werner

    2011-05-01

    High-performance liquid chromatography (HPLC) methods using a charged aerosol detector (CAD), a mass selective detector (MSD), and a diode array detector (DAD) were developed to characterize the nonionic surfactants d-α-tocopheryl polyethylene glycol (1000) succinate (TPGS) and Surfhope sugar ester D-1216 (sucrose laurate). The molecular structure and the heterogeneous composition resulting from different isomers and various lengths of polyethylene glycol (PEG) chains make it difficult to develop sensitive and specific analytical methods for both surfactants. Hence, there is lack of knowledge about the stability and grade of impurity of these compounds. Sucrose laurate does not possess any chromophore, thus UV detection is not applicable. Therefore, CAD and MSD have been used for determination. The aim of the study was to characterize these nonionic surfactants and to examine chemical stability at pH 1.0 and 37 °C, simulating harsh gastric conditions. It was shown that both compounds are liable to degradation under these conditions. Sucrose monolaurate exhibited a massive degradation within 8 h incubation due to cleavage of the glycosidic bondage. About 50% of sucrose monolaurate broke down, whereas a marginal amount of 3.4% (± 0.4%) of TPGS degraded into d-α-tocopheryl succinate and the associated PEG chain.

  13. Characterisation of betalain patterns of differently coloured inflorescences from Gomphrena globosa L. and Bougainvillea sp. by HPLC-DAD-ESI-MSn.

    Science.gov (United States)

    Kugler, Florian; Stintzing, Florian C; Carle, Reinhold

    2007-01-01

    In the present study, the betaxanthin (bx) and betacyanin patterns of differently coloured inflorescences from Gomphrena globosa L. and Bougainvillea sp. have been investigated in detail by applying reversed phase high-performance liquid chromatography-diode array detection (HPLC-DAD) coupled with positive ion electrospray mass spectrometry. Histidine-bx was found to be the predominant betaxanthin of Gomphrena globosa inflorescences. Furthermore, arginine-bx was detected as a novel betaxanthin, which to the best of our knowledge has not been reported as a pigment that occurs naturally so far. Dopa-bx was the major betaxanthin of Bougainvillea sp., although several minor betaxanthins were also present, including lysine-bx and putrescine-bx, novel betaxanthins hitherto not observed naturally. Remarkable differences in the betacyanin patterns between the purple, red and orange varieties were observed for both Gomphrena and Bougainvillea inflorescences. Hence, both the betacyanin profiles and the relative betaxanthin:betacyanin ratios determine the broad colour palette of Gomphrena petals and Bougainvillea bracts.

  14. Cleaning validation for residual estimation of olmesartan medoxomil on stainless steel surface of pharmaceutical manufacturing equipments using swab sampling and HPLC-DAD method

    Directory of Open Access Journals (Sweden)

    Nitin Dubey

    2013-06-01

    Full Text Available Prevention of cross contamination with active pharmaceutical ingredient is crucial and requires special attention in pharmaceutical industry. Current method validation describes residual determination of olmesartan medoxomil (OLME on stainless steel surface using swab sampling with a sensitive HPLC-DAD analysis. The acceptance limit was decided as 2 μg swab pro 100 cm2. Cotton swabs impregnated with extraction solution were used to determine residual drug content. Recoveries were 95.81%, 93.06%, and 96%. 24% with RSD below 1.5% at three concentration levels. Residual concentration was found to be linear in the range of 0.557–5.62 μg/mL, when estimated using Phenomenex Luna C18 (25 cm × 5 μm × 4.6 mm i.d. column at 1.0 mL/min flow rate at 258 nm. The mobile phase consisted of a mixture of acetonitrile: methanol: phosphate buffer pH 3.5: tetrahydrofuran (28:13:58:1 v/v/v/v. The LOD and LOQ for OLME were found to be 0.07 and 0.22 μg/mL, respectively. The validated method was found to be simple, selective and sensitive for demonstration of cleaning validation of OLME residues on the stainless steel surface.

  15. HPLC-DAD-MS Profiling of Polyphenols Responsible for the Yellow-Orange Color in Apple Juices of Different French Cider Apple Varieties.

    Science.gov (United States)

    Le Deun, Erell; Van der Werf, Remmelt; Le Bail, Gildas; Le Quéré, Jean-Michel; Guyot, Sylvain

    2015-09-09

    The pigments responsible for the yellow-orange coloration of apple juices have remained largely unknown up to now. Four French cider apple juices were produced in conditions similar to those used in the cider-making industry. The oxidized juices, characterized using the CIE L a b parameters, displayed various colors depending on the apple variety and native phenolic composition. HPLC-DAD-MS revealed contrasting pigment profiles related to oxidized tanning and nontanning molecules. The latter were divided into two groups according to their polarity and their visible spectra. With regard to phenolic classes, flavanol monomers and hydroxycinnamic acids played an essential role in the formation of oxidation products. Interestingly, dihydrochalcones appeared to include precursors of some yellow compounds. Indeed, the yellow pigment phloretin xyloglucoside oxidation product (PXGOPj), derived from phloretin xyloglucoside, was clearly identified in apple juices as a xyloglucose analogue of the yellow pigment phloridzin oxidation product (POPj), previously characterized in a model solution by Le Guernevé et al. (Tetrahedron Lett. 2004, 45 (35), 6673-6677).

  16. Identification of Phenolic Compounds in Red and Green Pistachio (Pistacia vera L.) Hulls (Exo- and Mesocarp) by HPLC-DAD-ESI-(HR)-MS(n).

    Science.gov (United States)

    Erşan, Sevcan; Güçlü Üstündağ, Özlem; Carle, Reinhold; Schweiggert, Ralf M

    2016-07-01

    Phenolic constituents of the nonlignified red and green pistachio hulls (exo- and mesocarp) were assessed by HPLC-DAD-ESI-MS(n) as well as by HR-MS. A total of 66 compounds was identified in the respective aqueous methanolic extracts. Among them, gallic acid, monogalloyl glucoside, monogalloyl quinic acid, penta-O-galloyl-β-d-glucose, hexagalloyl hexose, quercetin 3-O-galactoside, quercetin 3-O-glucoside, quercetin 3-O-glucuronide, and (17:1)-, (13:0)-, and (13:1)-anacardic acids were detected at highest signal intensity. The main difference between red and green hulls was the presence of anthocyanins in the former ones. Differently galloylated hydrolyzable tannins, anthocyanins, and minor anacardic acids were identified for the first time. Pistachio hulls were thus shown to be a source of structurally diverse and potentially bioactive phenolic compounds. They therefore represent a valuable byproduct of pistachio processing having potential for further utilization as raw material for the recovery of pharmaceutical, nutraceutical, and chemical products.

  17. An improved HPLC-DAD method for quantitative comparisons of triterpenes in Ganoderma lucidum and its five related species originating from Vietnam.

    Science.gov (United States)

    Ha, Do Thi; Loan, Le Thi; Hung, Tran Manh; Han, Le Vu Ngoc; Khoi, Nguyen Minh; Dung, Le Viet; Min, Byung Sun; Nguyen, Nguyen Phuong Dai

    2015-01-09

    An HPLC-DAD method for the quality control of wild and cultivated Ganoderma lucidum (Linhzhi) and related species samples was developed and validated. The quantitative determination of G. lucidum and its related species using 14 triterpene constituents, including nine ganoderma acids (compounds 4-12), four alcohols (compounds 13-16), and one sterol (ergosterol, 17) were reported. The standard curves were linear over the concentration range of 7.5-180 µg/mL. The LOD and LOQ values for the analyses varied from 0.34 to 1.41 µg/mL and from 1.01 to 4.23 µg/mL, respectively. The percentage recovery of each reference compound was found to be from 97.09% to 100.79%, and the RSD (%) was less than 2.35%. The precision and accuracy ranged from 0.81%-3.20% and 95.38%-102.19% for intra-day, and from 0.43%-3.67% and 96.63%-103.09% for inter-day, respectively. The study disclosed in detail significant differences between the quantities of analyzed compounds in different samples. The total triterpenes in wild Linhzhi samples were significantly higher than in cultivated ones. The total constituent contents of the five related Linhzhi samples were considerably lower than that in the G. lucidum specimens, except for G. australe as its constituent content outweighed wild Linhzhi's content by 4:1.

  18. Simultaneous determination of protocatechuic acid, syringin, chlorogenic acid, caffeic acid, liriodendrin and isofraxidin in Acanthopanax senticosus Harms by HPLC-DAD.

    Science.gov (United States)

    Li, Qing; Jia, Ying; Xu, Liang; Wang, Xiaohui; Shen, Zhenduo; Liu, Yulei; Bi, Kaishun

    2006-03-01

    A high performance liquid chromatography (HPLC) method was developed for the first time to quantify simultaneously the six major active ingredients in Acanthopanax senticosus (Rupr. et Maxim.) Harms, namely protocatechuic acid, syringin, chlorogenic acid, caffeic acid, liriodendrin and isofraxidin. The analysis was performed by a reverse phase gradient elution with an aqueous mobile phase (containing 0.05% phosphoric acid) modified by acetonitrile and diode-array multiple-wavelength UV detector (DAD). Six regression equations showed good linear relationships between the peak area of each marker and concentration. The recoveries of the markers listed above were 92.3%, 93.9%, 90.3%, 93.1%, 94.3% and 90.7%, respectively. The relative standard deviation of intra-day and inter-day were less than 2.7% and 3.1%, respectively. This method was validated for specificity, accuracy, precision and limits of quantification. Medicinal materials of ten commercial brands were analyzed and found to contain different amounts of the six bioactive markers. The method developed can be used for the quality control of Acanthopanax senticosus (Rupr. et Maxim.) Harms.

  19. Sesamin and sesamolin as unexpected contaminants in various cold-pressed plant oils: NP-HPLC/FLD/DAD and RP-UPLC-ESI/MS(n) study.

    Science.gov (United States)

    Górnaś, Paweł; Siger, Aleksander; Pugajeva, Iveta; Segliņa, Dalija

    2014-04-01

    Thirteen cold-pressed oils (Japanese quince seed, black caraway, flaxseed, rapeseed, hemp, peanut, sunflower, pumpkin, hazelnut, poppy, walnut, almond and sesame oil) manufactured by the same company over a 2-year period (2011-12) were assessed for lipophilic compounds. The presence of sesamin and sesamolin, two characteristic lignans of sesame oil, were detected in all tested plant oils. Both lignans were identified by NP-HPLC/FLD/DAD and confirmed by a RP-UPLC-ESI/MS(n) method. The lowest amount of sesamin and sesamolin was found for Japanese quince seed oil (0.10 and 0.27 mg/100 g), and the highest, excluding sesame oil, for almond oil (36.21 and 105.42 mg/100 g, respectively). The highly significant correlation between sesamolin and sesamin concentrations was found in all samples tested (r = 0.9999; p < 0.00001). These results indicate contamination of cold-pressed oils from the same source. This investigation highlights the fact that increasing the range of products manufactured by the same company can contribute to a lesser regard for the quality of the final product. Moreover, less attention paid to the quality of final product can be related to the health risks of consumers especially sensitive to allergens. Therefore, proper cleaning of processing equipment is needed to prevent cross-contact of cold-pressed oils.

  20. A Stability-Indicating HPLC-DAD Method for Determination of Ferulic Acid into Microparticles: Development, Validation, Forced Degradation, and Encapsulation Efficiency.

    Science.gov (United States)

    Nadal, Jessica Mendes; Toledo, Maria da Graça; Pupo, Yasmine Mendes; Padilha de Paula, Josiane; Farago, Paulo Vitor; Zanin, Sandra Maria Warumby

    2015-01-01

    A simple stability-indicating HPLC-DAD method was validated for the determination of ferulic acid (FA) in polymeric microparticles. Chromatographic conditions consisted of a RP C18 column (250 mm × 4.60 mm, 5 μm, 110 Å) using a mixture of methanol and water pH 3.0 (48 : 52 v/v) as mobile phase at a flow rate of 1.0 mL/min with UV detection at 320 nm. The developed method was validated as per ICH guidelines with respect to specificity, linearity, limit of quantification, limit of detection, accuracy, precision, and robustness provided suitable results regarding all parameters investigated. The calibration curve was linear in the concentration range of 10.0-70.0 μg/mL with a correlation coefficient >0.999. Precision (intraday and interday) was demonstrated by a relative standard deviation lower than 2.0%. Accuracy was assessed by the recovery test of FA from polymeric microparticles (99.02% to 100.73%). Specificity showed no interference from the components of polymeric microparticles or from the degradation products derived from acidic, basic, and photolytic conditions. In conclusion, the method is suitable to be applied to assay FA as bulk drug and into polymeric microparticles and can be used for studying its stability and degradation kinetics.

  1. A Stability-Indicating HPLC-DAD Method for Determination of Ferulic Acid into Microparticles: Development, Validation, Forced Degradation, and Encapsulation Efficiency

    Directory of Open Access Journals (Sweden)

    Jessica Mendes Nadal

    2015-01-01

    Full Text Available A simple stability-indicating HPLC-DAD method was validated for the determination of ferulic acid (FA in polymeric microparticles. Chromatographic conditions consisted of a RP C18 column (250 mm × 4.60 mm, 5 μm, 110 Å using a mixture of methanol and water pH 3.0 (48 : 52 v/v as mobile phase at a flow rate of 1.0 mL/min with UV detection at 320 nm. The developed method was validated as per ICH guidelines with respect to specificity, linearity, limit of quantification, limit of detection, accuracy, precision, and robustness provided suitable results regarding all parameters investigated. The calibration curve was linear in the concentration range of 10.0–70.0 μg/mL with a correlation coefficient >0.999. Precision (intraday and interday was demonstrated by a relative standard deviation lower than 2.0%. Accuracy was assessed by the recovery test of FA from polymeric microparticles (99.02% to 100.73%. Specificity showed no interference from the components of polymeric microparticles or from the degradation products derived from acidic, basic, and photolytic conditions. In conclusion, the method is suitable to be applied to assay FA as bulk drug and into polymeric microparticles and can be used for studying its stability and degradation kinetics.

  2. Residue and dissipation kinetics of thiamethoxam in a vegetable-field ecosystem using QuEChERS methodology combined with HPLC-DAD.

    Science.gov (United States)

    Abd-Alrahman, Sherif H

    2014-09-15

    The dissipation kinetics and residual levels of thiamethoxam in potato and soil under field ecosystem were determined using a QuEChERS method with HPLC-DAD. At fortification levels of 0.05, 0.1, 0.25, 0.5, 1.0 and 5.0 mg kg(-1), it was shown recovery was 99.4% (95.3-103.5%) for potato tubers, and 88.5% (86-91%) for soil with coefficient variation of the method (CV%) was less than 4% in potato tubers, and in soil less than 11%. For repeatability ranged from 1.27% to 4.77%. The LOD and LOQ were estimated to be 0.02 and 0.06 mg kg(-1), respectively. The half-lives were 2.92 and 1.4 days, respectively. The terminal residues of thiamethoxam were below the maximum residue limit (MRL 0.2 mg kg(-1)) after 6 days, which considered to be safe for human beings. These results contribute to establishing the scientific basis of the dosage of thiamethoxam for use in vegetable-field ecosystems.

  3. HPLC-DAD-ESI/MS Identification and Quantification of Phenolic Compounds in Ilex paraguariensis Beverages and On-Line Evaluation of Individual Antioxidant Activity

    Directory of Open Access Journals (Sweden)

    Delia B. Rodriguez-Amaya

    2013-03-01

    Full Text Available “Chimarrão” and “tererê” are maté (dried, toasted and milled Ilex paraguariensis leaves and stemlets beverages widely consumed in South America. This paper describes the application of HPLC-DAD-ESI/MS method for the identification and quantification of caffeoylquinic acids (CQA, flavonol glycosides and purine alkaloids in these beverages. The beverage samples were prepared from commercial lots of maté from Southern Brazil. The caffeoylquinic acids, 4,5-diCQA, 3-CQA, 5-CQA, and 4-CQA were the major compounds, having 238–289, 153–242, 183–263, and 123–188 μg/mL, respectively, for chimarrão and 206–265, 122–218, 164–209, 103–169 μg/mL, respectively, for tererê. Caffeine also had high amounts while glycosides of quercetin and kaempferol were found at much lower levels. The individual antioxidant activity was also determined by an on-line system that measured their ABTS•+ radical scavenging activity, showing that the antioxidant capacity was not proportional to the concentrations of the phenolic compounds. 3-CQA, quercetina-3-O-ramnosylglucoside, and quercetina-3-O-glucoside were the major contributors to the antioxidant capacity, although the quercetin glycosides had concentrations less than 10 times that of 3-CQA.

  4. Studies into the phenolic patterns of different tissues of pineapple (Ananas comosus [L.] Merr.) infructescence by HPLC-DAD-ESI-MS (n) and GC-MS analysis.

    Science.gov (United States)

    Steingass, Christof B; Glock, Mona P; Schweiggert, Ralf M; Carle, Reinhold

    2015-08-01

    In a comprehensive study, more than 60 phenolic compounds were detected in methanolic extracts from different tissues of pineapple infructescence by high-performance liquid chromatography with diode array detection and electrospray ionisation multiple-stage mass spectrometry (HPLC-DAD-ESI-MS (n) ) as well as by gas chromatography-mass spectrometry (GC-MS). The analytical workflow combining both methods revealed numerous compounds assigned for the first time as pineapple constituents by their mass fragmentations. Pineapple crown tissue was characterised by depsides of p-coumaric and ferulic acid. In contrast, major phenolic compounds in pineapple pulp extracts were assigned to diverse S-p-coumaryl, S-coniferyl and S-sinapyl derivatives of glutathione, N-L-γ-glutamyl-L-cysteine and L-cysteine, which were also identified in the peel. The latter was additionally characterised by elevated concentrations of p-coumaric, ferulic and caffeic acid depsides and glycerides, respectively. Two peel-specific cyanidin hexosides were found. Elevated concentrations of isomeric N,N'-diferuloylspermidines may be a useful tool for the detection of fraudulent peel usage for pineapple juice production. Mass fragmentation pathways of characteristic pineapple constituents are proposed, and their putative biological functions are discussed.

  5. Identification of new derivatives of 2-S-glutathionylcaftaric acid in aged white wines by HPLC-DAD-ESI-MS(n).

    Science.gov (United States)

    Cejudo-Bastante, María Jesús; Pérez-Coello, María Soledad; Hermosín-Gutiérrez, Isidro

    2010-11-10

    Glutathione, a natural occurring antioxidant, is a thiol-containing peptide present in grape must and wine. It is able to regenerate the o-diphenol group of enzymatically oxidized trans-caftaric acid, giving rise to 2-S-glutathionyl-trans-caftaric acid (also known as grape reaction product, GRP) and thus inhibiting the browning of wine. The amount of GRP present in a wine provides information on the oxidation history of the wine over its elaboration and aging. GRP has been proved to suffer hydrolysis in model solutions and wines. To know the actual content of glutathione involved in white wine browning inhibition as GRP, the GRP-derived products have been studied in 1-year-aged white wines by HPLC-DAD-ESI-MS(n). Online UV-vis spectra and pseudomolecular ions [(M + H)(+)] obtained by LC-MS supported the formation of some of the expected GRP hydrolysis products, mainly 2-S-glutathionyl-trans-caffeic acid (trans-GSCf), together with minor 2-S-(cysteinylglycyl)-trans-caftaric acid, 2-S-(γ-glutamylcysteinyl)-trans-caftaric acid, and 2-S-cysteinyl-trans-caftaric acid. On the basis of UV-vis and LC-MS(2) spectra, new GRP derivatives in aged white wines have been tentatively characterized for the first time as three monoethyl esters of GRP (GRP-Et) and also the cis isomers of GRP, GSCf, and some of the GRP-Et.

  6. HPLC-DAD-ESI/MS identification and quantification of phenolic compounds in Ilex paraguariensis beverages and on-line evaluation of individual antioxidant activity.

    Science.gov (United States)

    Peres, Renato G; Tonin, Fernando G; Tavares, Marina F M; Rodriguez-Amaya, Delia B

    2013-03-28

    "Chimarrão" and "tererê" are maté (dried, toasted and milled Ilex paraguariensis leaves and stemlets) beverages widely consumed in South America. This paper describes the application of HPLC-DAD-ESI/MS method for the identification and quantification of caffeoylquinic acids (CQA), flavonol glycosides and purine alkaloids in these beverages. The beverage samples were prepared from commercial lots of maté from Southern Brazil. The caffeoylquinic acids, 4,5-diCQA, 3-CQA, 5-CQA, and 4-CQA were the major compounds, having 238-289, 153-242, 183-263, and 123-188 μg/mL, respectively, for chimarrão and 206-265, 122-218, 164-209, 103-169 μg/mL, respectively, for tererê. Caffeine also had high amounts while glycosides of quercetin and kaempferol were found at much lower levels. The individual antioxidant activity was also determined by an on-line system that measured their ABTS•+ radical scavenging activity, showing that the antioxidant capacity was not proportional to the concentrations of the phenolic compounds. 3-CQA, quercetina-3-O-ramnosylglucoside, and quercetina-3-O-glucoside were the major contributors to the antioxidant capacity, although the quercetin glycosides had concentrations less than 10 times that of 3-CQA.

  7. Characterization and quantification of flavonoids and hydroxycinnamic acids in curly kale (Brassica oleracea L. Convar. acephala Var. sabellica) by HPLC-DAD-ESI-MSn.

    Science.gov (United States)

    Olsen, Helle; Aaby, Kjersti; Borge, Grethe Iren A

    2009-04-08

    Kale is a leafy green vegetable belonging to the Brassicaceae family, a group of vegetables including cabbage, broccoli, cauliflower, and Brussels sprouts, with a high content of health-promoting phytochemicals. The flavonoids and hydroxycinammic acids of curly kale ( Brassica oleracea L. ssp. oleracea convar. acephala (DC.) Alef. var. sabellica L.), a variety of kale, were characterized and identified primarily through HPLC-DAD-ESI-MS(n) analysis. Thirty-two phenolic compounds including glycosides of quercetin and kaempferol and derivatives of p-coumaric, ferulic, sinapic, and caffeic acid were tentatively identified, providing a more complete identification of phenolic compounds in curly kale than previously reported. Moreover, three hydroxycinnamic acids and one flavonoid with an unusual high grade of glycosylation, quercetin-3-disinapoyl-triglucoside-7-diglucoside, have been tentatively identified for the first time. The influence of different extraction conditions (extraction method, solvent type, solvent/solid ratio, and duration of extraction) was investigated. The total flavonol and hydroxycinnamic acid contents in curly kale determined as rutin equivalents (RE) were 646 and 204 mg of RE/100 g of fresh weight (fw), respectively. The contents of individual flavonoids ranged from 2 to 159 mg of RE/100 g of fw, with main compounds kaempferol-3-sinapoyl-diglucoside-7-diglucoside (18.7%) and quercetin-3-sinapoyl-diglucoside-7-diglucoside (16.5%). After acidic hydrolysis, two flavonol aglycones were identified in curly kale, quercetin and kaempferol, with total contents of 44 and 58 mg/100 g of fw, respectively.

  8. HPLC-DAD-ESI-MS(2) analytical profile of extracts obtained from purple sweet potato after green ultrasound-assisted extraction.

    Science.gov (United States)

    Zhu, Zhenzhou; Guan, Qingyan; Koubaa, Mohamed; Barba, Francisco J; Roohinejad, Shahin; Cravotto, Giancarlo; Yang, Xinsun; Li, Shuyi; He, Jingren

    2017-01-15

    Ultrasound pre-treatment (UAE) was applied to assist the extraction of valuable compounds (polyphenols (especially anthocyanins), and proteins) from purple sweet potato (PSP). Under optimum conditions (ultrasound time (40min); supplementary hot extraction (80°C) up to 120min; pH: 2.5; ethanol concentration: 58%), the highest concentrations of polyphenols (3.877mg/g), anthocyanins (0.293mg/g), and proteins (0.753mg/g) were found, with minimal specific energy consumption (8406J/mg). Moreover, anthocyanin and non-anthocyanin polyphenols in PSP extract from optimized extraction temperature were identified using HPLC-DAD-ESI-MS(2). The major identified anthocyanins were peonidin-3-caffeoyl-p-hydroxybenzoyl sophoroside-5-glucoside, peonidin-3-(6″-caffeoyl-6‴-feruloyl sophoroside)-5-glucoside, cyanidin-3-caffeoyl-p-hydroxybenzoyl sophoroside-5-glucoside, whereas the major identified non-anthocyanin molecules were quinic acid, chlorogenic acid, caffeic acid, and chlorogenic acid-3-glucose. The amount of the predominant anthocyanin and non-anthocyanin compounds from PSP extract obtained after UAE was higher than that extracted after conventional solvent extraction. The results obtained in this work demonstrated the efficiency of UAE for the recovery of anthocyanins from PSP.

  9. Highly sensitive HPLC-DAD method for the assay of gefitinib in patient plasma and cerebrospinal fluid: application to a blood-brain barrier penetration study.

    Science.gov (United States)

    Fang, Luo; Song, Yu; Weng, Xu; Li, Fanzhu; Xu, Yaping; Lin, Nengming

    2015-12-01

    The quantification of intracranial gefitinib (GEF) exposure is limited owing to the sensitivity of analytical equipment. Although mass spectrometry (MS) is the preferred method because of its high sensitivity, the equipment is not available in many laboratories, especially in developing Asian countries. In this paper, we developed a highly sensitive high performance liquid chromatography-diode array detector (HPLC-DAD) method for the assay of GEF in human cerebrospinal fluid (CSF) and plasma. GEF was extracted from CSF and plasma by solid-phase extraction and liquid-liquid extraction, respectively. The chromatographic separation was performed on a C18 column with gradient elution of 0.1% triethylamine solution and acetonitrile, then finally detected at 344 nm. This method was validated and proved to be highly sensitive with a lower limit of quantitation value of 0.11 ng/mL in CSF and 11 ng/mL in plasma. The blood-brain barrier penetration ratio of GEF ranged from 1.48 to 2.41%. This method provides a reliable MS-independent solution for the quantitation of GEF in patients' CSF and plasma.

  10. Identification and quantification of phenolic compounds of selected fruits from Madeira Island by HPLC-DAD-ESI-MS(n) and screening for their antioxidant activity.

    Science.gov (United States)

    Spínola, Vítor; Pinto, Joana; Castilho, Paula C

    2015-04-15

    Five fruits species commonly cultivated and consumed in Madeira Island (Portugal) were investigated for their phenolic profile by means of reversed phase high-performance liquid chromatography coupled to diode array detection and electrospray ionisation mass spectrometry (HPLC-DAD-ESI/MS(n)) and antioxidant potential. A large number of compounds were characterised, flavonoids and phenolic acids being the major components found in target samples, 39 compounds (flavonoids, phenolic acids, terpenoids, cyanogenic glycosides and organic acids) were identified in cherimoyas, lemons, papayas, passion-fruits and strawberries for the first time. Furthermore, all samples were systematically analysed for their total phenolic and flavonoid contents along with two radical scavenging methods (ABTS and ORAC) for antioxidant activity measurement. Target fruits presented high phenolic contents which is responsible for most of the antioxidant activity against radical reactive species (R(2)>0.80). Quantitative data showed that anthocyanins, in particular pelargonidin-3-O-hexoside (>300 mg/100 mL), present only in strawberries were the compounds in largest amounts but are the ones which contribute less to the antioxidant activity.

  11. Determination of tramadol hydrochloride in ampoule dosage forms by using UV spectrophotometric and HPLC-DAD methods in methanol and water media.

    Science.gov (United States)

    Küçük, Aysel; Kadioğlu, Yücel

    2005-02-01

    Two newly developed simple and sensitive methods for determination of tramadol hydrochloride in ampoule dosage forms were described and validated. Measurements for spectrophotometric method were performed using UV-Vis Spectrophotometer in ranges of 200-400 nm. The solutions of standard and the samples were prepared in methanol and water media and the UV absorption spectrums of tramadol were monitored with maximum absorptions at 275 and 271 nm for both mediums, respectively. The standard calibration curves of tramadol were constructed by plotting absorbance vs. concentration in the concentration range with the final dilution of 10-100 microg ml-1. Reversed phase chromatography for HPLC method was conducted using a Phenomenex Bondclone C18 column with an isocratic mobile phase consisting of 25% acetonitrile in 75% 0.01 M phosphate buffer (pH 3). The effluent was monitored on a DAD detector at 218 nm. Linear response (r>0.99) was observed over the range of 0.5-40 microg ml-1 for methanol and water and run on six different occasions. The methods were applied successfully to pharmaceutical ampoule forms, but also for comparison in two different solvent media. Besides, it was completely validated and proven to be rugged.

  12. Development and validation of carbofuran and 3-hydroxycarbofuran analysis by high-pressure liquid chromatography with diode array detector (HPLC-DAD) for forensic Veterinary Medicine.

    Science.gov (United States)

    Gonçalves, Vagner; Hazarbassanov, Nicolle Queiroz; de Siqueira, Adriana; Florio, Jorge Camilo; Ciscato, Claudia Helena Pastor; Maiorka, Paulo Cesar; Fukushima, André Rinaldi; de Souza Spinosa, Helenice

    2017-09-14

    Agricultural pesticides used with the criminal intent to intoxicate domestic and wild animals are a serious concern in Veterinary Medicine. In order to identify the pesticide carbofuran and its metabolite 3- hydroxycarbofuran in animals suspected of exogenous intoxication a high pressure liquid chromatography with diode array detector (HPLC-DAD) method was developed and validated in stomach contents, liver, vitreous humor and blood. The method was evaluated using biological samples from seven different animal species. The following parameters of analytical validation were evaluated: linearity, precision, accuracy, selectivity, recovery and matrix effect. The method was linear at the range of 6.25-100μg/mL and the correlation coefficient (r(2)) values were >0.9811 for all matrices. The precision and accuracy of the method was determined by coefficient of variation (CV) and the relative standard deviation error (RSE), and both were less than 15%. Recovery ranged from 74.29 to 100.1% for carbofuran and from 64.72 to 100.61% for 3-hydroxycarbofuran. There were no significant interfering peaks or matrix effects. This method was suitable for detecting 25 positive cases for carbofuran amongst a total of 64 animal samples suspected of poisoning brought to the Toxicology Diagnostic Laboratory, School of Veterinary Medicine and Animal Sciences, University of Sao Paulo. Copyright © 2017 Elsevier B.V. All rights reserved.

  13. Characterization of Flavonoids in the Ethomedicine Fordiae Cauliflorae Radix and Its Adulterant Millettiae Pulchrae Radix by HPLC-DAD-ESI-IT-TOF-MSn

    Directory of Open Access Journals (Sweden)

    Lan-Lan Fan

    2013-12-01

    Full Text Available Fordiae Cauliflorae Radix (FC, the root of Fordia cauliflora Hemsl and Millettiae Pulchrae Radix [MP, the root of Millettia pulchra (Benth. Kurz var. laxior (Dunn Z. Wei], which go under the same local name of “Daluosan”, have long been used in Southern China for the treatment of stroke, paralysis, dementia in children, Alzheimer’s disease and other diseases. The same local name and similar functions always confuse users. To further utilize these two ethnodrugs and identify them unambiguously, an HPLC-DAD-ESI-IT-TOF-MSn method was developed to separate and characterize the flavonoids in FC and MP. A total of 41 flavonoids were detected, of which six compounds were identified by comparing their retention time and MS data with those of the reference standards, and the others were tentatively identified based on their tandem mass spectrometry data obtained in the positive ion detection mode. Nineteen of these characterized compounds are reported from these two plants for the first time.

  14. 千里香枝中39个多甲氧基黄酮的HPLC-DAD-ESI-MS/MS分析%Characterization of thirty-nine polymethoxylated flavonoids (PMFs) in the branches of Murraya paniculata by HPLC-DAD-ESI-MS/MS

    Institute of Scientific and Technical Information of China (English)

    张加余; 卢建秋; 高晓燕; 张倩; 李宁; 屠鹏飞; 乔延江

    2013-01-01

    AIM:To investigate the polymethoxylated flavonoids (PMFs) in the branches of Murraya paniculata (L.) Jack.METHODS:A sensitive HPLC-DAD-ESI-MS/MS method was established to screen PMFs in the branches of M.paniculata based on the analysis of six PMF standards in the positive mode by CID-MS/MS.RESULTS:The diagnostic fragmentation pathways for polymethoxylated flavones,polymethoxylated flavanones,polymethoxylated chalcones and PMF glycosides were summarized,respectively.According to the MS fragmentation pathways,39 PMFs,including 24 flavones,10 flavanones or chalcones and 5 PMFs glycosides were screened.CONCLUSION:The results indicated that the developed analytical method could be employed as a rapid,effective technique for the chemical screening of PMFs in TCMs extracts.%目的:定性分析千里香枝中的多甲氧基黄酮类成分.方法:建立一种灵敏的HPLC-DAD-ESI-MS/MS方法,筛选千里香枝中的多甲氧基黄酮.结果:分别研究总结了多甲氧黄酮、二氢黄酮、查耳酮以及多甲氧基黄酮苷的诊断性断裂途径.结合这些特征和EIC-MS/MS实验,筛选了39个多甲氧基黄酮类成分,包括24个多甲氧基黄酮、10个多甲氧基二氢黄酮或查耳酮、5个多甲氧基黄酮苷.结论:本研究为从复杂物质体系中筛选多甲氧基黄酮提供了一种快速有效的方法.

  15. Chemical Characteristics Combined with Bioactivity for Comprehensive Evaluation of Tumuxiang Based on HPLC-DAD and Multivariate Statistical Methods

    Institute of Scientific and Technical Information of China (English)

    Xia Gao; Yu-Ling Ma; Pei Zhang; Xiao-Ping Zheng; Bo-Lu Sun; Fang-Di Hu

    2016-01-01

    Background: The dried roots of Inula helenium L. (IH) and Inula racemosa Hook f. (IR) are used commonly as folk medicine under the name of “tumuxiang (TMX)”. Phenolic acid compounds and their derivatives, as main active constituents in IH and IR, exhibit prominent anti-inflammation effect. Objective: To develop a holistic method based on chemical characteristic and anti-inflammation effect for systematically evaluating the quality of twenty-seven TMX samples (including 18 IH samples and 9 IR samples) from different origins. Methods: HPLC fingerprints data of AL (Aucklandia lappa Decne.) whose dried root was similar with HR was added for classification analysis. The HPLC fingerprints of twenty-seven TMX samples and four AL samples were evaluated using hierarchical clustering analysis (HCA) and principle component analysis (PCA). The spectrum-efficacy model between HPLC fingerprints and anti-inflammatory activities was investigated by principal component regression (PCR) and partial least squares(PLS). Results: All samples were successfully divided into three main clusters and peaks 7, 9, 11, 22, 24 and 26 had a primary contribution to classify these medicinal herbs. The results were in accord with the appraisal results of herbs. The spectrum-efficacy relationship results indicated that citric acid, quinic acid, caffeic acid-β-D-glucopyranoside, chlorogenic acid, caffeic acid, 1,3-O-dicaffeoyl quinic acid, tianshic acid and 3β-Hydroxypterondontic acid had main contribution to anti-inflammatory activities. Conclusion: This comprehensive strategy was successfully used for identification of IH, IR and AL, which provided a reliable and adequate theoretical basis for the bioactivity relevant quality standards and studying the material basis of anti-inflammatory effect of TMX.

  16. HPLC-DAD和HPLC-ESI-MS/MS检测小麦及土壤中申嗪霉素残留方法比较%Determination of Phenazine-1-carboxylic Acid by HPLC-DAD and HPLC-ESI-MS/MS: A Comparison of the Two Methods

    Institute of Scientific and Technical Information of China (English)

    刘钰; 冯义志; 刘伟

    2013-01-01

    [目的]选出更优化的检测小麦、麦苗/麦秆和土壤中申嗪霉素的残留分析方法.[方法]样品经提取、净化后,分别采用DAD检测器和质谱检测器检测.[结果]申嗪霉素在小麦、麦苗/麦秆和土壤中平均添加回收率分别为80.3%~95.8%、71.7%~94.9%和77.2%~101.1%,RSD分别为1.59%~8.03%、1.74%~8.40%和1.29%~8.55%.申嗪霉素用DAD和MS/MS两种方法在小麦、麦苗/麦秆和土壤中LOQ分别为0.01、0.05、0.005 mg/kg和0.005、0.01、0.005 mg/kg.[结论]HPLC-ESI-MS/MS方法操作简便、灵敏度高,更适合检测田间样品中申嗪霉素残留.%[Aims] To select an analytical method for determining phenazine-1-carboxylic acid residues in wheat, seedling/ straw and soil. [Methods] The sample was extracted and purified, followed by DAD detector and HPLC-MS/MS analysis, respectively. [Results] The results showed the average recoveries of phenazine-1-carboxylic acid were 80.3-95.8% in wheat, 71.7-94.9% in seeding/straw and 77.2-101.1% in soil, with relative standard deviations were 1.59-8.03%, 1.74-8.40% and 1.29-8.55%, respectively. The limits of quantification (LOQ) of HPLC-DAD and HPLC-ESI-MS/MS in wheat, seedling/straw and soil were 0.01, 0.05, 0.005 mg/kg and 0.005, 0.01, 0.005 mg/kg, respectively. [Conclusions] Comparison of the two methods, HPLC-ESI-MS/MS was sample, high sensitivity and appropriate for determining phenazine-1-carboxylic acid residue from field samples

  17. FISH BILIARY POLYCYCLIC AROMATIC HYDROCARBON METABOLITES ESTIMATED BY FIXED-WAVELENGTH FLUORESCENCE: COMPARISON WITH HPLC-FLUORESCENT DETECTION

    Science.gov (United States)

    Fixed wavelength fluorescence (FF) was compared to high-performance liquid chromatography with fluorescence detection (HPLC-F) as an estimation of polycyclic aromatic hydrocarbon (PAH) exposure to fish. Two excitation/emission wavelength pairs were used to measure naphthalene- an...

  18. Chromatographic fingerprint analysis and simultaneous determination of eight lignans in Justicia procumbens and its compound preparation by HPLC-DAD.

    Science.gov (United States)

    Wang, Linan; Pan, Jianyu; Yang, Meihua; Wu, Jun; Yang, Junshan

    2011-03-01

    HPLC fingerprints were developed for the quality evaluation of Justicia procumbens and its compound preparation, Jian-er syrup, together with the simultaneous quantification of eight arylnaphthalide lignans (6'-hydroxy justicidin B, 6'-hydroxy justicidin A, 6'-hydroxy justicidin C, justicidin B, chinensinaphthol methyl ether, justicidin C, taiwanin C, and neojusticin A). Samples were separated with a Shiseido Capcell Pak C(18) reversed-phase column (250×4.6 mm id, 5 μm) using acetonitrile and water as the mobile phase. The column temperature was maintained at 35°C and the wavelength of detector was set at 256 nm. For fingerprint analysis, 17 peaks were selected as the characteristic peaks for the evaluation of the similarities among different J. procumbens samples collected in different places. The structures of lignans were confirmed by diagnostic fragments in the positive ESI-MS(n) . The new method was successfully applied for the chromatographic fingerprint analysis and simultaneous determination of eight lignans in its compound preparation, Jian-er syrup. All the results indicated that HPLC fingerprint assay in combination with multi-marker determination afforded a useful method for the quality control of J. procumbens and its compound preparation, Jian-er syrup.

  19. Multivariate curve resolution-assisted determination of pseudoephedrine and methamphetamine by HPLC-DAD in water samples.

    Science.gov (United States)

    Vosough, Maryam; Mohamedian, Hadi; Salemi, Amir; Baheri, Tahmineh

    2015-02-01

    In the present study, a simple strategy based on solid-phase extraction (SPE) with a cation exchange sorbent (Finisterre SCX) followed by fast high-performance liquid chromatography (HPLC) with diode array detection coupled with chemometrics tools has been proposed for the determination of methamphetamine and pseudoephedrine in ground water and river water. At first, the HPLC and SPE conditions were optimized and the analytical performance of the method was determined. In the case of ground water, determination of analytes was successfully performed through univariate calibration curves. For river water sample, multivariate curve resolution and alternating least squares was implemented and the second-order advantage was achieved in samples containing uncalibrated interferences and uncorrected background signals. The calibration curves showed good linearity (r(2) > 0.994).The limits of detection for pseudoephedrine and methamphetamine were 0.06 and 0.08 μg/L and the average recovery values were 104.7 and 102.3% in river water, respectively.

  20. Determination of Water Extracts of Melastoma Candidum and Melastoma Normale by HPLC-DAD-ESI-MS/MS%野牡丹和羊开口水提液中化学成分的HPLC-DAD-ESI-MS/MS分析

    Institute of Scientific and Technical Information of China (English)

    唐铁鑫; 郑恒; 刘燕; 邱新华

    2016-01-01

    目的:采用高效液相色谱-电喷雾串联质谱法对野牡丹和羊开口水提液中化学成分进行分析比较。方法用反相 C18色谱柱进行成分分离,以甲醇(含10mM甲酸铵)-水(含10mM甲酸铵)梯度洗脱,二级管阵列检测器和电喷雾电离串联质谱在线同时检测。结果野牡丹和羊开口水提液的化学成分基本一致,均推定鉴定出含有没食子酸和(表)儿茶素,并且均主要检测出没食子酸衍生物和(表)儿茶素衍生物。结论两者水提液的主要成分均为没食子酸及其衍生物、(表)儿茶素及其衍生物,而野牡丹和羊开口传统用药均采用水煎剂,这表明两者功能与主治基本一致的原因是它们的水提液化学成分基本一致。%OBJECTIVE The chemical composition of water extract of Melastoma candidum and Melastoma nor-male were determined by liquid chromatography-electrospray ionization tandem mass spectrometry.METHODS The high performance liquid chromatography was equipped with a reversed phase C18 column.A gradient binary mobile phase system was consisted of methanol ( containing 10 mM ammonium formate) and water ( containing 10 mM am-monium formate).Online detection were carried out by DAD and ESI-MS/MS at the same time.RESULTS The chemical composition of water extract of Melastoma candidum and Melastoma normale were basically the same.Gallic acid and ( epi) catechin were tentatively identified in water extracts of both of them,and gallic acid derivatives and (epi) catechin derivatives were the main compounds determined in water extracts of both of them.CONCLUSION Gallic acid and its derivatives,(epi) catechin and its derivatives were the main water-soluble components of bothplants.Moreover,in traditional practice both plants were used in water decoction.It indicated that the reason that thefunction and indications of both plants are basically the same derives from the fact that their water

  1. HPLC-DAD法检测染红中药中20种非法添加色素%Simultaneous Determination of 20 Pigments in Illegally Dyed Chinese Herbs by HPLC-DAD

    Institute of Scientific and Technical Information of China (English)

    李启艳; 朱日然; 孙萍; 姜国萍

    2015-01-01

    目的:建立高效液相色谱-二极管阵列检测器( HPLC-DAD)法同时测定常见染红中药中非法添加的20种人工合成色素。方法采用70%甲醇超声提取样品,通过Agilent TC-C18反相色谱柱(4.6 mm×250 mm,5μm)分离,以甲醇-0.02 mol·L-1醋酸铵溶液为流动相进行梯度洗脱,于210~650 nm波长范围用DAD检测,用保留时间结合待测物的紫外吸收光谱验证。结果同时分离并检测出20种常用红色及橙色系色素,市售西红花、朱砂中检出酸性橙域及808猩红。结论该方法可同时检测常见染红中药中常用的非法添加人工合成色素,样品处理简便,方法快速,结果准确可靠,重复性好。%Objective To develop a HPLC-DAD method for the simultaneous determination of 20 synthetic pigments in dyed Traditional Chinese Medicine. Methods The samples were extracted with ultrasonic by 70% methanol, separated on a Agilent TC-C18 column ( 4. 6 mmí250 mm, 5 μm ) by gradient elution using a mobile phase made up of methanol and 0. 02 mol·L-1 ammonium acetate, and detected at 210-650 nm. The retention times and ultraviolet absorption spectrums were used to text and verify. Results The twenty red and orange synthetic pigments were separated and detected. OrangeIIsodium salt and 808 scarlet were detected in some batch of Croci stigma and Cinnabaris. Conclusion The method is simple, rapid, accurate, repeatable for determining twenty synthetic pigments in dyed Traditional Chinese Medicine.

  2. MCR-ALS APLICADO NO MONITORAMENTO QUANTITATIVO DO PROCESSO DE ELETRODEGRADAÇÃO DA ATRAZINA USANDO ESPECTROS UV: RESULTADOS COMPARATIVOS COM HPLC-DAD COMO UM MÉTODO DE REFERÊNCIA

    Directory of Open Access Journals (Sweden)

    Thálisson S. Souza

    2016-02-01

    Full Text Available Electrodegradation of atrazine in water was performed using homemade (PA and PB and purchased (PC boron-doped diamond anodes. The degradation was monitored off-line by analyzing total organic carbon and high performance liquid chromatography with diode array detector (HPLC-DAD and at-line by UV spectroscopy. The spectra were recorded every 2 min. The rank deficiency problem was resolved by assembling an augmented column-wise matrix. HPLC was employed to separate the original and byproducts degradation components. Aiming the same goal, multivariate curve resolution - alternating least squares (MCR-ALS was applied to resolve the UV spectroscopic data. Comparison between HPLC and MCR-ALS separations is presented. By using MCR-ALS the spectra of atrazine and two byproducts were successfully resolved and the resulted concentration profiles properly represented the system studied. The ALS explained variance (R2 for PA, PB and PC was equal to 99.99% for all of them and the lack of fit for PA, PB and PC were 0.39, 0.34 and 0.54 respectively. The correlation (R between the recovered and pure spectra were calculate for each electrodegradation, validating the MCR-ALS results. The average R was equal to 0.997. The spectral and concentration profiles described with this new approach are in agreement with HPLC-DAD results. The proposed method is an alternative to classical analyses for monitoring of the degradation process, mainly due to the simplicity, fast results and economy.

  3. My dad

    Institute of Scientific and Technical Information of China (English)

    夏文慧

    2011-01-01

    My dad is forty years old. He is short. He has big eyes Beause he didn't study hard when he was a child. Now he has to teach himself. He is strict with my liltle brother and me. I don't like him. I often say he is a "bad" dad. On Sundays he leaves me at home alone. If I go out to play with my friends. He will beat me. Then he keeps me doing my homework. He doesn't let me watch TV. He doesn't let me spend any money. There are many things about my dad.

  4. An Improved HPLC-DAD Method for Quantitative Comparisons of Triterpenes in Ganoderma lucidum and Its Five Related Species Originating from Vietnam

    Directory of Open Access Journals (Sweden)

    Do Thi Ha

    2015-01-01

    Full Text Available An HPLC-DAD method for the quality control of wild and cultivated Ganoderma lucidum (Linhzhi and related species samples was developed and validated. The quantitative determination of G. lucidum and its related species using 14 triterpene constituents, including nine ganoderma acids (compounds 4–12, four alcohols (compounds 13–16, and one sterol (ergosterol, 17 were reported. The standard curves were linear over the concentration range of 7.5–180 µg/mL. The LOD and LOQ values for the analyses varied from 0.34 to 1.41 µg/mL and from 1.01 to 4.23 µg/mL, respectively. The percentage recovery of each reference compound was found to be from 97.09% to 100.79%, and the RSD (% was less than 2.35%. The precision and accuracy ranged from 0.81%–3.20% and 95.38%–102.19% for intra-day, and from 0.43%–3.67% and 96.63%–103.09% for inter-day, respectively. The study disclosed in detail significant differences between the quantities of analyzed compounds in different samples. The total triterpenes in wild Linhzhi samples were significantly higher than in cultivated ones. The total constituent contents of the five related Linhzhi samples were considerably lower than that in the G. lucidum specimens, except for G. australe as its constituent content outweighed wild Linhzhi’s content by 4:1.

  5. Comparison of the Sesquiterpene Ester Profiles of Different Specialized Armillariella mellea Mycelia Using HPLC-DAD Chromatography%蜜环菌不同特化菌体的HPLC-DAD图谱比较

    Institute of Scientific and Technical Information of China (English)

    程显好; 王春兰; 郭顺星

    2006-01-01

    采用麦芽汁液体培养基培养蜜环菌获得菌丝体,采用麦芽汁半固体培养基培养蜜环菌获得蜜环菌菌索、皮壳状菌丝和子实体.以高效液相色谱-二极管阵列检测法(High Performance Liquid Chromatography-Diode Array detector,HPLC-DAD)比较四种菌体的甲醇提取物的成分差异.液体培养的蜜环菌菌丝体与固体培养的蜜环菌菌索、皮壳状菌丝和子实体的蜜环菌素类成分有明显差别.图谱分析表明,菌索与皮壳状菌丝的成分接近,与子实体和液体培养菌丝体之间有较大差别.

  6. Rapid characterization of 96 chemical constituents in Citri Reticulatae Folium (leaves of ‘Fuju’) using HPLC-DAD-ESI-MSn%HPLC-DAD-ESI-MSn技术对橘叶(福橘叶)中96个化学成分的快速分离和鉴定

    Institute of Scientific and Technical Information of China (English)

    曹规划; 付庆荣; 张藏曼; 王弘; 陈世忠

    2016-01-01

    为了对橘叶(福橘叶)中的化学成分进行较为系统地研究,我们首次采用液质联用技术((HPLC-DAD-ESI-MSn)系统地分离和鉴定了橘叶中的化学成分.最终,我们在橘叶中解析了96个化学成分,包括:31个酚酸类化合物,4个黄酮苷元,6个黄酮单氧苷,10个黄酮双氧苷,5个黄酮单碳苷和5个黄酮双碳苷化合物,以及6个黄酮碳氧苷,5个3-羟基-3-甲基戊二酰基取代的黄酮苷,1个黄烷-3-醇和2个生物碱类化合物,并且初步鉴定出了21个多甲基黄酮类化合物(PMFs).在上述解析的96个化合物中,52个化合物为首次在橘叶中发现,15个化合物为首次在柑橘属植物中发现.同时,我们在橘叶中也发现了9个潜在的新化合物.%In order to systematically investigate the chemical constituents of Citri Reticulatae Folium (leaves of ‘Fuju’),an analytical method that included high-performance liquid chromatography,diode array detection,electrospray ionization,and ion-trap time-of-flight mass spectrometry (HPLC-DAD-ESI-MSn) was used to separate and identify the individual chemical components of Citri Reticulatae Folium.As a result,96 compounds were tentatively identified in this study:including 31 phenolic acids,4 flavonoid aglycones,6 flavonoid mono-O-glycosides,10 flavonoid-O-diglycosides,5 flavonoid mono-C-glycosides,5 flavonoid di-C-glycosides,6 flavonoid O,C-glycosides,5 (3-hydroxy-3-methylglutaryl) glycosyl flavonoids,1 flavan-3-ol,and 2 alkaloids.In addition,21 polymethoxy flavonoids (PMFs) were identified in this paper.Among these compounds,52 compounds,which were previously found in other Citrus plants,have been identified for the first time in Citri Reticulatae Folium.15 compounds have not been previously found in the Citrus genus were identified.Moreover,9 potentially new compounds have also been detected in this paper.This is the first report of the full characterization of chemical components of Citri Reticulatae Folium (leaves of

  7. Combined use of liquid chromatography-hybrid quadrupole time-of-flight mass spectrometry (LC-QTOF-MS) and high performance liquid chromatography with photodiode array detector (HPLC-DAD) in systematic toxicological analysis.

    Science.gov (United States)

    Broecker, Sebastian; Pragst, Fritz; Bakdash, Abdulsallam; Herre, Sieglinde; Tsokos, Michael

    2011-10-10

    Time of flight mass spectrometry provides new possibilities of substance identification by determination of the molecular formula from accurate molecular mass and isotope pattern. However, the huge number of possible isomers requires additional evidence. As a suitable way for routine performance of systematic toxicological analysis, a method for combined use of liquid chromatography-hybrid quadrupole time-of-flight mass spectrometry (LC-QTOF-MS) and high performance liquid chromatography with diode array detector (HPLC-DAD) was developed and applied to blood samples from 77 death cases. The blood samples were prepared by extraction with CH(2)Cl(2) and by protein precipitation with acetonitrile (1:4 (v/v)). The evaporated extracts were reconstituted in 35% acetonitril/0.1% formic acid/H(2)O and aliquots were injected for analysis by LC-QTOF-MS (Agilent 6530) and HPLC-DAD (Agilent 1200). A valve switching system enabled simultaneous operation of both separated chromatographic lines under their respective optimal conditions using the same autosampler. The ESI-QTOF-MS instrument was run in data dependent acquisition mode with switching between MS and MS/MS (cycle time 1.1s) and measuring the full mass spectra and the collision induced dissociation (CID) fragment spectra of all essential [M+H](+) ions. Libraries of accurate mass CID spectra (~2500 substances) and of DAD-UV spectra (~3300 substances) of the authors were used for substance identification. The application of this procedure is demonstrated in detail at four examples with multiple drug intake or administration. In the 77 cases altogether 198 substances were identified (87 by DAD and 195 by QTOF-MS) with a frequency between 1 and 20. In practical application, the sample preparation proved to be suitable for both techniques and for a wide variety of substances with different polarity. The automatic performance of the measurements was efficient and robust. Mutual confirmation, decrease of false positive and

  8. Determinação de 25-hidroxivitamina D2 e D3 em plasma por CLAE-DAD Determination of 25-hidroxy-vitamin D2 and D3 in plasma by HPLC-DAD

    Directory of Open Access Journals (Sweden)

    Daiana Manuele Kich

    2012-10-01

    concentration of 25-hidroxy-vitamin D (25(OHD in the fractions D2 (25(OHD2 and D3 (25(OHD3. OBJECTIVE: To develop an analytical method using high-performance liquid chromatography with diode-array detection (HPLC-DAD for the determination of 25(OHD2 and 25(OHD3 in plasma. MATERIALS AND METHODS: 25(OHD3 and 25(OHD2 were extracted from plasma samples with hexane and dodecaphenone was used as internal standard. The separation was performed in an ACE 5 C18 column, with particle size of 5 µm (4,6 × 150 mm, mobile phase methanol-water (80:20, v/v and quantification at 265 nm. RESULTS: Accuracy was in the range of 98.4 to 107.5%. Intra-assay precision was between 6.5 and 9.2% for 25(OHD3 and 3.7 and 8.7 for 25(OHD2. Inter-assay precision was between 2.9 and 6% for 25(OHD3 and 4 and 4.5 for 25(OHD2. The limit of quantification was 10 ng/l. Concentrations of 25(OHD3 in samples from 32 elderly patients were between 10.1 and 32.4 ng/ml, characterizing vitamin D deficiency in this group. DISCUSSION: The method allowed the quantification of 25(OHD2 and 25(OHD3. Furthermore, the sample preparation was relatively simple and fast. The method was selective with an adequate separation of metabolites and internal standard with no interfering substances. CONCLUSION: Not only did the developed method show suitable analytical performance, but it may also be applied in clinical conditions.

  9. Simultaneous Determination of Neomycin Sulfate and Hydrochloric Dyclonine in Compound Twain Oint-ments by HPLC-DAD-ELSD%HPLC-DAD-ELSD法同时测定复方吐温软膏中硫酸新霉素与盐酸达克罗宁的含量

    Institute of Scientific and Technical Information of China (English)

    段松冷; 曾蔚欣; 孙路路

    2015-01-01

    目的::建立同时测定复方吐温软膏中硫酸新霉素与盐酸达克罗宁含量的HPLC-DAD-ELSD法。方法:采用 Agilent ZOR BAXSB-C18色谱柱(250 mm ×4.6 mm,5μm),以含0.1 mol·L-1三氟乙酸的水溶液、乙腈为流动相,梯度洗脱,流速为1.0 ml·min-1,DAD的检测波长为282 nm,ELSD的漂移管温度为50℃,雾化温度为60℃,载气流速为1.6 L·min-1,柱温为35℃。结果:硫酸新霉素检测浓度的线性范围为141.540~323.520μg·ml-1(r=0.9996),平均回收率为98.87%,RSD=0.95%(n=9);盐酸达克罗宁检测浓的线性范围为28.000~64.000μg·ml-1(r=0.9996),平均回收率为99.57%,RSD=1.10%(n=9)。结论:该方法结果准确、灵敏度高、重复性好,在同一色谱条件下实现了复方吐温软膏中全部有效成分的含量测定,为其质量控制提供了依据。%Objective:To establish an HPLC-DAD-ELSD method for the simultaneous determination of neomycin sulfate and hy-drochloric dyclonine in compound Twaln ointments. Methods:The assay was performed on an Agilent ZOR BAXSB-C18 column(250 mm × 4. 6 mm, 5μm) with acetonitrile-water as the mobile phase with gradient elution at a flow rate of 1. 0 ml·min-1 . The detection wavelength of DAD was 282 nm. The evaporator temperature of ELSD was set at 50℃ and the nebulizer temperature was set at 60℃with the gas flow rate of 1. 6 L·min-1 . The column temperature was kept at 35℃. Results:The linear range of neomycin sulfate was 141. 54-323. 52 μg·mL-1(r=0. 999 6) with the average recovery of 98. 87%(RSD=0. 95%, n=9). The linear range of hydro-chloric dyclonine was 28. 00-64. 00 μg·mL-1(r=0. 999 6) with the average recovery of 99. 57%(RSD=1. 10%, n=9). Conclu-sion:The method is accurate, sensitive and reproducible, and under the same chromatographic conditions, the determination of all the active ingredients in compound Twaln ointments is achieved, which provides basis for the quality control.

  10. HPLC-DAD-MS/MS Analysis of Purine Alkaloids and Tea Polyphenols in Young Leaves of Yinghong 1, Yinghong 9 and Qimen Cultivars%英红1号、英红9号和祁门茶树芽叶中嘌呤生物碱和茶多酚的HPLC-DAD-MS/MS分析

    Institute of Scientific and Technical Information of China (English)

    卢嘉丽; 王冬梅; 苗爱清; 杨得坡; 陈建萍

    2009-01-01

    采用HPLC-DAD-MS/MS技术对英红1号、英红9号和祁门茶树的芽叶进行了分析,鉴定出了2种嘌呤生物碱,7种儿茶素类化合物以及2种非儿茶素类茶多酚.对这3个茶树品种的嘌呤生物碱和儿茶素类成分质量分数进行比较分析,结果表明三者的儿茶素类总质量分数均大于20%,其中英红1号和祁门红茶品种在儿茶素类成分的组成上较为接近,EGCG质量分数均为最高;而英红9号质量分数最高的是ECG.

  11. Identification and Quantification of Common Adulterants of Saffron (Crocus sativus) by HPLC-DAD%西红花常见伪品的HPLC-DAD鉴别和掺伪量测定

    Institute of Scientific and Technical Information of China (English)

    安慧景; 张继; 孙兰; 南垚; 武佳; 周立东

    2012-01-01

    目的:建立HPLC-DAD方法,一次性鉴别西红花常见伪品红花、菊花,并测定其掺伪量.方法:采用高效液相色谱法,流动相为乙腈-0.3%磷酸水;波长范围200~410 nm;羟基红花黄色素A检测波长402 nm,绿原酸检测波长327 nm;梯度洗脱.结果:通过HPLC分析,确定羟基红花黄色素A、绿原酸分别为红花、菊花的特征成分,测定样品含量,可鉴别出样品中是否掺入了红花、菊花,并确定伪品掺入量.羟基红花黄色素A 0.005~0.25 mg·mL-1,绿原酸0.001~0.05 mg·mL-1范围内线性关系良好,方法回收率高,重复性好.结论:此方法能一次性鉴别西红花常见伪品红花、菊花,并测定其掺伪量,对其它药材的相关研究也有参考价值.%This study was aimed to develop a method to identify and quantify common adulterants of saffron by simultaneously determining specific constituents by HPLC-DAD. The separation was achieved using a C18 column and CH3CN-0.3%H3PO4 solution as a mobile phase with gradient elution. Hydroxysafflor yellow A and chloro-genic acid, used as specific constituents, were detected at 402 nm and 327 nm, respectively. By detecting specific constituents, we can find out if samples were adulterated with safflower and/or chrysanthemum and calculate the percentage. The method was validated in terms of linearity, precision, accuracy and recovery. And results were found to be satisfactory. This method is simple, sensitive and operational in the identification and quantification of common adulterants of saffron.

  12. Application of HPLC-DAD after SPE/QuEChERS with ZrO2-based sorbent in d-SPE clean-up step for pesticide analysis in edible oils.

    Science.gov (United States)

    Tuzimski, Tomasz; Rejczak, Tomasz

    2016-01-01

    In this study, the solid-phase extraction/quick, easy, cheap, effective, rugged and safe (SPE/QuEChERS) technique was adapted to develop a simple sample treatment for multi-residue pesticide analysis of edible oils. The proposed method is based on liquid-liquid partitioning with acetonitrile followed by dispersive solid phase extraction using zirconia-coated silica particles for extract purification. To evaluate the described method, 21 pesticides belonging to different chemical classes were analysed using high performance liquid chromatography with diode-array detection (HPLC-DAD). For validation purposes, recovery studies were performed at 75 ng g(-1), 125 ng g(-1), 250 ng g(-1), 500 ng g(-1) and 1000 ng g(-1) levels. Recoveries were over the range of 50-130% for most of the analytes, with relative standard deviations less than 15% being observed. HPLC-DAD provided suitable linearity, precision and accuracy. The validated method was successfully applied to the analysis of edible oil samples selected from the market.

  13. QuEChERS-based extraction with dispersive solid phase extraction clean-up using PSA and ZrO2-based sorbents for determination of pesticides in bovine milk samples by HPLC-DAD.

    Science.gov (United States)

    Rejczak, Tomasz; Tuzimski, Tomasz

    2017-02-15

    In this study, a quick, easy, cheap, effective, rugged and safe (QuEChERS) extraction technique was adapted to develop a simple sample treatment for multi-residue pesticide analysis in milk samples. The proposed method is based on liquid-liquid partitioning with acetonitrile followed by dispersive solid phase extraction clean-up using primary secondary amine along with zirconia-coated silica particles for extract purification. Identification and quantification of 30 pesticides was conducted via high performance liquid chromatography with diode-array detection (HPLC-DAD). Recoveries were from 70 to 100% for the vast majority of the analytes, with relative standard deviations less than 20% being observed. HPLC-DAD provided suitable linearity, precision and accuracy. For 28 of 30 analytes in the study method limit of quantification values (mLOQs) comply with the most recent European Union guidelines for the maximum residue levels (MRLs) in milk. Negligible matrix effect was observed due to efficient extract clean-up with ZrO2-based sorbents.

  14. Determination of acetylsalicylic acid and salicylic acid in foods, using HPLC with fluorescence detection.

    NARCIS (Netherlands)

    Venema, D.P.; Hollman, P.C.H.; Janssen, P.L.T.M.K.; Katan, M.B.

    1996-01-01

    We developed a specific and sensitive HPLC method with fluorescence detection for the determination of free acetylsalicylic acid, free salicylic acid, and free salicylic acid plus salicylic acid after alkaline hydrolysis (free-plus-bound) in foods. Acetylsalicylic acid was detected after postcolumn

  15. Determination of acetylsalicylic acid and salicylic acid in foods, using HPLC with fluorescence detection.

    NARCIS (Netherlands)

    Venema, D.P.; Hollman, P.C.H.; Janssen, P.L.T.M.K.; Katan, M.B.

    1996-01-01

    We developed a specific and sensitive HPLC method with fluorescence detection for the determination of free acetylsalicylic acid, free salicylic acid, and free salicylic acid plus salicylic acid after alkaline hydrolysis (free-plus-bound) in foods. Acetylsalicylic acid was detected after postcolumn

  16. Screening and identification of multiple components in Tanreqing injection using RP-HPLC combined with DAD and ESI-TOF/MS.

    Science.gov (United States)

    Liu, Shao-Yong; Xue, Dong-Sheng; Pan, Jian-Chao; Zhang, Wen-Ming; Li, Wen-Long; Qu, Hai-Bin

    2014-07-01

    A liquid chromatography coupled with diode array detector (DAD) and electrospray ionization time-of-flight mass spectrometry (ESI-TOF/MS) method was developed for the screening and identification of the multiple components in Tanreqing injection, a well-known Chinese medicine injection in China. By combining the DAD spectrum and the accurate mass measurement of ESI-TOF/MS, twelve components in Tanreqing injection were identified. This study contributes to clarifying the nature of Tanreqing injection, and provides an effective and reliable process for the comprehensive and systematic characterization of complex traditional Chinese medicine preparations.

  17. Quantification of nortriptyline in plasma by HPLC and fluorescence detection.

    Science.gov (United States)

    Almudever, Patricia; Peris, José-Esteban; Garrigues, Teresa; Diez, Octavio; Melero, Ana; Alós, Manuel

    2010-03-15

    A simple, sensitive and specific high-performance liquid chromatography method has been developed for the determination of nortriptyline (NT) in plasma samples. The assay involved derivatization with 9H-fluoren-9-ylmethyl chloroformate (Fmoc-Cl) and isocratic reversed-phase (C(18)) chromatography with fluorescence detection. The developed method required only 100 microl of plasma sample, deproteinized and derivatized in one step. Calibration curves were lineal over the concentration range of 5-5000 ng/ml. The derivatization reaction was performed at room temperature in 20 min and the obtained NT derivative was stable for at least 48 h at room temperature. The within-day and between-day relative standard deviation was below 8%. The limit of detection (LOD) was 2 ng/ml, and the lower limit of quantification (LLOQ) was established at 10 ng/ml. The method was applied on plasma collected from rats, at different time intervals, after intravenous administration of 0.5 mg of NT.

  18. A Stability-Indicating HPLC-DAD Method for Determination of Stiripentol: Development, Validation, Kinetics, Structure Elucidation and Application to Commercial Dosage Form

    OpenAIRE

    Hany W. Darwish; Abdelhameed, Ali S; Mohamed I. Attia; Bakheit, Ahmed H.; Khalil, Nasr Y; Al-Majed, Abdulrahman A.

    2014-01-01

    A rapid, simple, sensitive, and accurate isocratic reversed-phase stability-indicating high performance liquid chromatography method has been developed and validated for the determination of stiripentol and its degradation product in its bulk form and pharmaceutical dosage form. Chromatographic separation was achieved on a Symmetry C18 column and quantification was achieved using photodiode array detector (DAD). The method was validated in accordance with the ICH requirements showing specific...

  19. Application of an efficient strategy based on MAE, HPLC-DAD-MS/MS and HSCCC for the rapid extraction, identification, separation and purification of flavonoids from Fructus Aurantii Immaturus.

    Science.gov (United States)

    Wang, Chen; Pan, Yaju; Fan, Guorong; Chai, Yifeng; Wu, Yutian

    2010-03-01

    This study presents an efficient strategy based on microwave-assisted extraction (MAE), HPLC-DAD-MS/MS and high-speed counter-current chromatography (HSCCC) for the rapid extraction, identification, separation and purification of active components from the traditional Chinese medicine Fructus Aurantii Immaturus. An LC-DAD-MS/MS method was applied for the screening and structural identification of main components in crude extract, and five components were preliminarily identified as neoeriocitrin, narirutin, naringin, hesperidin and neohesperidin according to their UV and mass spectra. An efficient MAE method for the extraction of the three most abundant components (narirutin, naringin and neohesperidin) was optimized by the combination of univariate and multivariate approaches. The crude extract was then separated and purified by HSCCC and a total of 61.6 mg of narirutin, 207.3 mg of naringin and 159.5 mg of neohesperidin at high purities of 98.1, 97.2 and 99.5%, respectively, were obtained from 1.42 g of crude extract. The recoveries of these compounds were 86, 93 and 89%, respectively.

  20. Determination of acetylsalicylic acid and salicylic acid in foods, using HPLC with fluorescence detection.

    OpenAIRE

    Venema, D.P.; Hollman, P.C.H.; Janssen, P.L.T.M.K.; Katan, M B

    1996-01-01

    We developed a specific and sensitive HPLC method with fluorescence detection for the determination of free acetylsalicylic acid, free salicylic acid, and free salicylic acid plus salicylic acid after alkaline hydrolysis (free-plus-bound) in foods. Acetylsalicylic acid was detected after postcolumn hydrolysis to salicylic acid. With the method for free acetylsalicylic acid and salicylic acid, recovery was 95-98␏or acetylsalicylic acid added to foods and 92-102␏or salicylic acid. Recovery of a...

  1. Separation and identification of phenolic compounds of extra virgin olive oil from Olea europaea L. by HPLC-DAD-SPE-NMR/MS. Identification of a new diastereoisomer of the aldehydic form of oleuropein aglycone.

    Science.gov (United States)

    Pérez-Trujillo, Míriam; Gómez-Caravaca, Ana María; Segura-Carretero, Antonio; Fernández-Gutiérrez, Alberto; Parella, Teodor

    2010-08-25

    The phenolic fraction of a monovarietal extra virgin olive oil (EVOO) from Olea europaea L. var. Cornezuelo was studied by the hyphenated HPLC-DAD-SPE-NMR/MS techniques. This survey led to the identification of 25 main compounds. One was identified as a new diastereoisomer of the aldehydic form of oleuropein aglycone (AOA) and characterized by 1D and 2D NMR techniques. The relative configuration of this new AOA was determined as 5R*,8S*,9S* on the basis of the results obtained from the combination of NOE experiments and Monte Carlo conformational search calculations. Assuming, as for the described diastereoisomers, that the new AOA comes from the natural oleuropein aglycone (OA), the absolute configuration was proposed as 5S,8R,9R.

  2. Response surface optimization of microwave-assisted extraction for HPLC-fluorescence determination of puerarin and daidz

    Institute of Scientific and Technical Information of China (English)

    Ying-Kun Liu; E Yan; Han-Ying Zhan; Zhi-Qi Zhang

    2011-01-01

    Microwave-assisted extraction was optimized with response surface methodology for HPLC-fluorescence determination of puerarin and daidzein in Radix Puerariae thomsonii.The optimized extraction procedure was achieved by soaking the sample with 70% methanol

  3. 高效液相色谱-质谱检测红葡萄皮中花色苷%HPLC - DAD - MS - MS determination of anthocyanins in red grape skin

    Institute of Scientific and Technical Information of China (English)

    季梅; 李强; 娄红祥

    2011-01-01

    Objective:To establish an HPLC - DAD - MS - MS method for determination of anthocyanins from red grape skin. Methods;The major anthocyanins of grape skin was determined by HPLC - DAD - MS - MS method. AichromBond C18 column (250 mm x4. 6 mm,5 μm) was used in HPLC assay. Mobile phase A and B were 2% formic acid and acetonitrile,respectively. 0 min:10% B;10 min:25% B;15 min:30% B;35 min:50% B. The flow rate was 0. 6 mL·min-1. The detective wavelength was 530 nm. The PDA detector and negative ion mode of MS were adopted. Results; 15 anthocyanins were determined from the red grape skin of Mountain Daze. Conclusion;It is sensitive and selective to determine anthocyanins of grape skin with HPLC - MS method. The method is also accurate and reliable.%目的:建立高效液相色谱-质谱联用(LC -MS)方法,检测红葡萄皮中花色苷.方法:采用AichromBond C18色谱柱(250mm×4.6 mm,5μm),流动相A(2%甲酸水溶液)和B(乙腈)梯度洗脱(0min∶10%B;10 min:25% B;15 min∶30% B;35 min∶50%B).流速:0.6 mL· min-1,检测波长:530 nm,二极管阵列检测器,质谱检测设定为负离子模式.结果:大泽山玫瑰香红葡萄皮中主要花色苷有15种.结论:LC - MS法检测红葡萄皮中花色苷,具有高的灵敏度和选择性,方法准确可靠.

  4. HPLC- DAD method for simultaneous determination of five constituents in Qizhi Weitong granules%HPLC-DAD法同时测定气滞胃痛颗粒中5个成分的含量

    Institute of Scientific and Technical Information of China (English)

    张赟华; 董媛; 李忠琼; 刘莹莹; 张雯洁

    2012-01-01

    Objective : To establish an HPLC method for simultaneous determination of five constituents (albiflorin, paeoniflorin, liquiritin, naringin, neohesperidin) in Qizhi Weitong granules. Methods: The separation was performed on an Elite Hypersil C18 column(4. 6 mm × 150 mm,5 μm)with the gradient elution of acetonitrile ( A) -water(B) (0 -45 min,10% A→28% A)at a flow rate of 1. 0 mL ·min-1. The DAD detection wavelength was 200 -400 nm. The column temperature was 30 °C. Results; The contents of albiflorin and paeoniflorin were determined at 230 nm wavelength with good linearity in the range of 0. 12 - 6. 14μg and 0. 40 - 7. 92 μg, respectively; the content of liquiritin was determined at 237 nm wavelength with good linearity in the range of 0. 06-3. 18μg;The contents of naringin and neohesperidin were determined at 283 nm wavelength with good linearity in the range of 0. 20 - 9. 84 μg and 0. 12 - 6. 14 μg, respectively. The average recoveries of above five constituents were 98. 1% ,98. 3% , 98. 9% ,98. 6% and 97. 1%,with RSD of 0. 63% ,0. 69% ,0. 96% ,1. 5% and 1. 3% Respectively. Conclusion; The developed method is rapid, simple, and accurate, which is helpful for the quality control of Qizhi Weitong granules.%目的:建立HPLC法同时测定气滞胃痛颗粒中白芍药苷、芍药苷、甘草苷、柚皮苷和新橙皮苷的含量.方法:采用依利特Hypersil C18色谱柱(4.6mm×150mm,5μm),以乙腈(A)-水(B)为流动相梯度洗脱(0~ 45 min,10%A→28%A),流速1.0mL· min-1,采用DAD检测器在200~400 nm波长处进行检测,柱温30℃.结果:调取不同波长的色谱图分别计算各成分的含量,白芍药苷( 230 nm)、芍药苷(230 nm)、甘草苷(237 nm)、柚皮苷(283 nm)和新橙皮苷(283 nm)进样量分别在0.12~6.14 μg(r=0.9998),0.40 ~ 7.92μg(r=0.9998),0.06 ~ 3.18 μg(r=1.000),0.20 ~9.84 μg(r=0.9999),0.12~6.14μg(r =0.9999)范围内呈现良好的线性关系.平均回收率(n=6)分别为98.1%,98.3%,98.9%,98

  5. Evaluation of the effect of TM208 on the activity of five cytochrome P450 enzymes using on-line solid-phase extraction HPLC-DAD: a cocktail approach.

    Science.gov (United States)

    Lin, Wensi; Zhang, Jianmei; Ling, Xiaomei; Yu, Ning; Li, Jing; Yang, Haisong; Li, Runtao; Cui, Jingrong

    2013-04-01

    A rapid, simple, and sensitive on-line solid-phase extraction HPLC-DAD method for simultaneous evaluation of the activity of five CYP450 isoforms (CYP1A2, CYP2C19, CYP2D6, CYP2E1 and CYP3A4) in vivo has been developed and validated. The five specific probe substrates include caffeine (1A2), metoprolol (2D6), dapsone (3A4), omeprazole (2C19) and chlorzoxazone (2E1). Automated pre-purification of plasma and enrichment of analytes were performed using a C18 on-line solid-phase extraction cartridge. After being eluted from the cartridge, the analytes and the internal standard antipyrine were separated on a C18 RP analytical column and analyzed by DAD. The method was validated to quantify the concentration ranges of 0.05-50.0 μg/ml for dapsone and omeprazole, 0.1-50.0 μg/ml for caffeine and 0.2-50.0 μg/ml for metoprolol and chlorzoxazone. The linearity (R(2)) for all analytes tested was exceeded 0.99. The intra-day precision ranged from 0.29 to 13% and the inter-day precision ranged from 5.0 to 15%, respectively. The intra-day and inter-day accuracy were between 86.7% and 113.6%. The extraction recoveries were in the range 82.8-109.9% for all the analytes and internal standard antipyrine. This method was successfully applied to evaluate the effects of TM208 on rat five CYP450 isoforms.

  6. Quantification of protein thiols using ThioGlo 1 fluorescent derivatives and HPLC separation.

    Science.gov (United States)

    Hoff, Signe; Larsen, Flemming H; Andersen, Mogens L; Lund, Marianne N

    2013-04-07

    A method for quantification of total soluble protein-derived thiols in beer was developed based on the formation of fluorescent adducts with the maleimide compound ThioGlo 1. The problem of interference from fluorescent adducts of sulfite and ThioGlo 1 was solved by HPLC separation of the adducts followed by fluorescence detection. Using standard addition of GSH, a detection limit of 0.028 μM thiols was achieved. The application and validation of the method was demonstrated for beers with different color intensities, and the application range is in principle for any biological system containing thiols. However, the quantification of cysteine was complicated by a lower fluorescence response of its ThioGlo 1 adducts. Based on the studies of the responses of a series of cysteine-derived thiols and (1)H NMR studies of the structures of ThioGlo 1 adducts with GSH and cysteine, it was concluded that thiols with a neighboring free amino group yield ThioGlo 1 adducts with a reduced fluorescence intensity.

  7. HPLC- DAD determination of 5 alkaloids in commercial available Kushen's rhizome and root%HPLC-DAD法测定商品苦参根茎和根中5个生物碱的含量

    Institute of Scientific and Technical Information of China (English)

    张萍; 韩南银; 杨璐; 王耀欣; 殷其蕾; 贺锐锐; 詹先王; 刘勇; 袁丽佳; 李中杰

    2011-01-01

    Objective:To determine the respective content of matrine, oxymatrin, sophocarpine, oxysophocarpine and sophoridine in commercial Kushen's rhizomes and roots so as to find out if the total contents of matrine and oxymatrin meet the request of not less than 1% in ChP( Vol. 1)2010 Ed. Methods:The procedure was performed on a C18(4. 6 mm X 150 mm,5 μm) column with a gradient elution of mobile phase A( 10 mmol·L-1 ammonium acetate aqueous,adjusted to pH 9. 2 with ammonia water) and mobile phase B(acetonitrile). The flow rate was 1 mL· min-1 and the detective wavelength of DAD was set at 220 nm. Results;The linear ranges of matrine,oxymatrin,sophocarpine, oxysophocarpine and sophoridine were 0. 1575 - 2. 205μg (r =0. 9999 ) , 0. 8985 - 12. 58μg ( r = 0. 9999) ,0. 1360 - 1. 904μg(r = 0. 9999),0. 4398 - 6. 153μg(r = 0. 9998), and 0. 2092 - 2. 926μg (r = 0. 9998), respectively;The recoveries( n = 3) were 101. 0% - 102. 5% ( RSD≤2. 1 % ), 100. 8% - 101. 6% (RSD sg2. 8% ), 102. 3% - 103. 6% ( RSD ≤1. 2% ), 102. 2% - 103.4% ( RSD≤1. 1% ), and 102. 4% - 103. 3% (RSD ≤2.9% ) ,respectively. Conclusion:The HPLC method shows a good precision and accuracy. The total contents of matrine and oxymatrine in both Kushen's rhizomes and roots are not less than 1. 0% , which meet thestan-dardsin ChP( Vol. 1)2010 Ed.%目的:测定商品苦参根茎和根中苦参碱、氧化苦参碱、槐果碱、氧化槐果碱、槐定碱的含量,主要了解苦参根茎中苦参碱和氧化苦参碱总量是否达到中国药典2010年版规定的不低于1.0%.方法:采用Welch Materials XtimateTMC18(4.6 mm×150mm,5μm)色谱柱,流动相A为10 mmol·L-1醋酸铵水溶液(0.1%氨水调至pH9.2),流动相B为乙腈,梯度洗脱,流速1 mL·min-1,检测波长220 nm.结果:苦参碱进样量在0.1575~2.205 μg(r=0.9999),氧化苦参碱在0.8985~12.58 μg(r=0.9999),槐果碱在0.1360 ~1.904 μg(r =0.9999),氧化槐果碱在0.4398~6.153 μg(r=0.9998),槐定碱在0.2092~2.926

  8. Concentrations of Nicotinamide in Plasma by RP-HPLC With Fluorescence Detection

    Directory of Open Access Journals (Sweden)

    Pan Zhipeng

    2016-01-01

    Full Text Available The purpose of this study is to establish a new method for detecting nicotinamide concentration in plasma. In the experiment, the high performance liquid chromatography (HPLC method was used, with a fluorescence detector. The nicotinamide in the plasma was first converted to N1- methylnicotinamide, then reacted with acetophenone under certain conditions to produce fluorescent derivatives for testing. The method is a kind of highly sensitive detection, of which the lower limit is 10 ng/mL, the recovery rate is between 92.75% and 105.13%, and the relative standard deviation (RSD is between 3.76% and 4.43%. The results showed that this measurement method is accurate, sensitive and rapid. It meets the requirements of the experiment, and applies to the detection of nicotinamide concentration in plasma.

  9. Development of a triple hyphenated HPLC-radical scavenging detection-DAD-SPE-NMR system for the rapid identification of antioxidants in complex plant extracts

    NARCIS (Netherlands)

    Pukalskas, A.; Beek, van T.A.; Waard, de P.

    2005-01-01

    A rapid method for the simultaneous detection and identification of radical scavenging compounds in plant extracts was developed by combining an HPLC with on-line radical scavenging using DPPH as a model radical and an HPLC¿DAD¿SPE¿NMR system. Using this method a commercial rosemary extract was inve

  10. A novel fluorescence derivatization method combined with HPLC for determining the activities of endogenous caspase.

    Science.gov (United States)

    Liu, Jiachi; Lu, Ye; Liang, Jianying

    2012-11-07

    A novel fluorescence derivatization method combined with HPLC was developed to detect the activity of caspase-3 and -8 in two cell lines (Hela cells and A549 cells) which were activated by low temperature-assisted ultraviolet irradiation (LT-UV), mitomycin C (MMC) and camptothecin during the apoptosis, respectively. Two peptide substrates for either caspase-3 or -8 were designed, of which peptide fragments were obtained by enzymatic modification, followed by fluorescence derivatization. A single fluorescent product was formed when a peptide was heated at 120 °C for 10 min in a neutral aqueous medium (pH 7.0) containing catechol, sodium periodate and sodium borate. Commercial kits for detecting the activity of caspase-3 and -8 were used as a control. The relative activity of the caspases detected by fluorescence derivatization was similar to that obtained by commercial kits, which indicated that the novel method is reliable. The activity assays of recombinant human caspases showed that the novel method provided higher selectivity than that of commercial kits, which proved it to be more accurate for determining the activity of caspases in apoptosis.

  11. Application of HPLC Combined with Laser Induced Fluorescence for Protein Profile Analysis of Tissue Homogenates in Cervical Cancer

    OpenAIRE

    Sujatha Bhat; Ajeetkumar Patil; Lavanya Rai; Kartha, V. B.; Santhosh Chidangil

    2012-01-01

    A highly objective method, High Performance Liquid Chromatography with Laser Induced Fluorescence (HPLC-LIF) technique was used to study the protein profiles of normal and cervical cancer tissue homogenates. A total of 44 samples including normal cervical biopsy samples from the hysterectomy patients and the patients suffering from different stages of the cervical cancer were recorded by HPLC-LIF and analysed by Principle Component Analysis (PCA) to get statistical information on different t...

  12. A Stability-Indicating HPLC-DAD Method for Determination of Stiripentol: Development, Validation, Kinetics, Structure Elucidation and Application to Commercial Dosage Form.

    Science.gov (United States)

    Darwish, Hany W; Abdelhameed, Ali S; Attia, Mohamed I; Bakheit, Ahmed H; Khalil, Nasr Y; Al-Majed, Abdulrahman A

    2014-01-01

    A rapid, simple, sensitive, and accurate isocratic reversed-phase stability-indicating high performance liquid chromatography method has been developed and validated for the determination of stiripentol and its degradation product in its bulk form and pharmaceutical dosage form. Chromatographic separation was achieved on a Symmetry C18 column and quantification was achieved using photodiode array detector (DAD). The method was validated in accordance with the ICH requirements showing specificity, linearity (r (2) = 0.9996, range of 1-25 μg/mL), precision (relative standard deviation lower than 2%), accuracy (mean recovery 100.08 ± 1.73), limits of detection and quantitation (LOD = 0.024 and LOQ = 0.081 μg/mL), and robustness. Stiripentol was subjected to various stress conditions and it has shown marked stability under alkaline hydrolytic stress conditions, thermal, oxidative, and photolytic conditions. Stiripentol degraded only under acidic conditions, forming a single degradation product which was well resolved from the pure drug with significantly different retention time values. This degradation product was characterized by (1)H-NMR and (13)C-NMR spectroscopy as well as ion trap mass spectrometry. The results demonstrated that the method would have a great value when applied in quality control and stability studies for stiripentol.

  13. A Stability-Indicating HPLC-DAD Method for Determination of Stiripentol: Development, Validation, Kinetics, Structure Elucidation and Application to Commercial Dosage Form

    Directory of Open Access Journals (Sweden)

    Hany W. Darwish

    2014-01-01

    Full Text Available A rapid, simple, sensitive, and accurate isocratic reversed-phase stability-indicating high performance liquid chromatography method has been developed and validated for the determination of stiripentol and its degradation product in its bulk form and pharmaceutical dosage form. Chromatographic separation was achieved on a Symmetry C18 column and quantification was achieved using photodiode array detector (DAD. The method was validated in accordance with the ICH requirements showing specificity, linearity (r2=0.9996, range of 1–25 μg/mL, precision (relative standard deviation lower than 2%, accuracy (mean recovery 100.08±1.73, limits of detection and quantitation (LOD = 0.024 and LOQ = 0.081 μg/mL, and robustness. Stiripentol was subjected to various stress conditions and it has shown marked stability under alkaline hydrolytic stress conditions, thermal, oxidative, and photolytic conditions. Stiripentol degraded only under acidic conditions, forming a single degradation product which was well resolved from the pure drug with significantly different retention time values. This degradation product was characterized by 1H-NMR and 13C-NMR spectroscopy as well as ion trap mass spectrometry. The results demonstrated that the method would have a great value when applied in quality control and stability studies for stiripentol.

  14. Determination of Amphetamine, Amfepramone and Fenproporex in Urine Samples by HPLC-DAD: Application to a Population of Brazilian Truck Drivers

    OpenAIRE

    Takitane,Juliana; Almeida, Rafael M.; Tiago F. de Oliveira; Prado,Natanael V.; Muñoz,Daniel R.; Leyton,Vilma; Yonamine, Mauricio

    2016-01-01

    Commercially available immunoassay tests are designed to detect the presence of amphetamine/methamphetamine or methylenodioxyamphetamines. However, it is known that Brazilian truck drivers also report the use of other illicit amphetamines, such as amfepramone and fenproporex. Thus, a method was developed and validated in order to quantify amphetamine-type stimulants (amphetamine, fenproporex and amfepramone) in urine by high performance liquid chromatography with diode array detection (HPLC-D...

  15. Rapid finding and quantification of the major antioxidant in water extracts of three marine drug organisms from China by online HPLC-DAD/MS-DPPH.

    Science.gov (United States)

    Zheng, Li; Chen, Junhui; Zhao, Hengqiang; Shi, Qian; Yang, Baijuan; Cheng, Hongyan; Zang, Jiaye; Wang, Xiaoru

    2012-01-01

    A method based on high-performance liquid chromatography (HPLC) with diode array detector coupled with electrospray ionisation-mass spectrometry and an online detection system for radical scavenging was established and used to rapidly find and quantify antioxidant compounds in the water extracts of Hippocampus japonicus Kaup, Hippocampus kuda Bleeker and Syngnathus acus Linnaeus. The online screening results revealed the presence of one major radical scavenging compound identified as hypoxanthine by comparison of mass data and retention time with the standard. Subsequently, the developed HPLC method was applied to quantify hypoxanthine in different H. japonicus, H. kuda and S. acus samples. The results indicated that the developed HPLC method is simple and reliable for the quantification of hypoxanthine with a detection limit at 0.002 µg  mL(-1), and a high recovery from 96.3% to 102.1%. This method provides a powerful tool for rapid identification and quantification of free radical scavenging compounds in complex marine natural products.

  16. Supercritical fluid extraction for some carbamate insecticides in potatoes samples, with HPLC/fluorescence determination and HPLC/mass spectrometry confirmation

    OpenAIRE

    Gilvanda Silva Nunes; Teresa Cristina R. Santos; Damiá Barceló; Alexandre Santos Pimenta; Maria Lúcia Ribeiro

    2002-01-01

    Six supercritical fluid extraction (SFE) methods were tested, by varying the following operational parameters: CO2 pressure, time and temperature of extraction, type and proportion of static modifier, and Hydromatrix®/sample rate into cell. Firstly, insecticide carbamates were extracted from spiked potatoes samples (fortification level of 0,5 mg.Kg-1) by using SPE procedures, and then final extracts were analyzed HPLC/fluorescence. Good performance was observed with SFE methods that operated ...

  17. Determination of lignans in edible and nonedible parts of pomegranate (Punica granatum L.) and products derived therefrom, particularly focusing on the quantitation of isolariciresinol using HPLC-DAD-ESI/MSn.

    Science.gov (United States)

    Fischer, Ulrike Anna; Jaksch, Ariane Verena; Carle, Reinhold; Kammerer, Dietmar Rolf

    2012-01-11

    A method for the characterization and quantitation of phyto-estrogenic lignans from pomegranate (Punica granatum L.) fruits and fruit-derived products by HPLC-DAD-MS(n) was developed. For this purpose, edible and nonedible parts of pomegranate (aril, peel, mesocarp, seed, and twigs), commercial juices, juices produced on pilot-plant scale, and encapsulated dietary supplements were analyzed. In addition to the peel, mesocarp, and twigs, lignans were detected in two juices obtained from entire fruits, four commercial juices, and three encapsulated pomegranate extracts. Isolariciresinol was the predominant lignan with contents of 5.0, 10.5, and 45.8 mg/kg dry matter in processed pomegranate mesocarp, peel, and twigs, respectively. In contrast, due to their low amounts, quantitation of lignans in pomegranate products was impossible. Therefore, contrary to previous assumptions, lignans were found to be less relevant in pomegranate-derived products. However, the byproduct from pomegranate processing may be used for lignan extraction. The method presented allows one to differentiate between pomegranate-derived products obtained from fruits without peels or by dejuicing applying low pressures, which were devoid of lignans, and those obtained from entire fruits applying high pressures, thus containing lignans. Consequently, this study helps to optimize process technology aiming at the recovery of preparations with well-desired compositions, which may reduce the risk of a wide range of diseases, such as certain types of cancer.

  18. Identification of C-geranylated flavonoids from Paulownia catalpifolia Gong Tong fruits by HPLC-DAD-ESI-MS/MS and their anti-aging effects on 2BS cells induced by H2O2.

    Science.gov (United States)

    Tang, Wen-Zhao; Wang, Ying-Ai; Gao, Tian-Yang; Wang, Xiao-Jing; Zhao, Yun-Xue

    2017-05-01

    The fruits of Paulownia catalpifolia Gong Tong are used as a Chinese folk herbal medicine for the treatment of enteritis, tonsillitis, bronchitis, and dysentery, etc. Our previous study has identified new C-geranylated flavanones with obvious anti-proliferative effects in lung cancer A549 cells. In the present study, a new C-geranylated flavone, paucatalinone C (1) and five known C-geranylated flavanones (2-6) were isolated. In addition, a total of 34 C-geranylated flavonoids were detected by HPLC-DAD-ESI-MS/MS coupling techniques from the CH2Cl2 extract of P. catalpifolia. Futhermore, anti-aging effects of isolated compounds were evaluated in vitro with premature senescent 2BS cells induced by H2O2. Phytochemical results indicated that P. catalpifolia was a natural resource of abundant C-geranylated flavonoids. Diplacone (3) and paucatalinone A (5) were the potent anti-aging agents in the premature senescent 2BS cells induced by H2O2 and the C-geranyl substituent may be an important factor because of its lipophilic character. Copyright © 2017 China Pharmaceutical University. Published by Elsevier B.V. All rights reserved.

  19. Determinação do perfil fitoquímico de extrato com atividade antioxidante da espécie medicinal Cordia verbenacea DC. por HPLC-DAD

    Directory of Open Access Journals (Sweden)

    M.M. Santi

    2014-06-01

    Full Text Available O presente trabalho teve por objetivo investigar a atividade antioxidante dos extratos das folhas de Cordia verbenacea obtido por maceração em etanol e partição em solventes orgânicos. O infuso das folhas também foi investigado. O teor de fenóis totais foi avaliado pelo método de Folin-Ciocalteau e o de flavonoides totais pela formação de complexo com cloreto de alumínio. O extrato etanólico, as subfrações e o infuso foram testados em diversas concentrações para determinar a atividade sequestradora de DPPH expressa em termos de sua CE50. A melhor atividade antioxidante encontrada foi para o extrato em acetato de etila, EA, CE50 15,0 ± 0,5 µg.mL-1. Os ensaios espectrofotométricos revelaram altas concentrações de fenóis e de flavonoides no extrato EA. A análise por HPLC-DAD foi realizada para se obter o perfil de UV-Vis dos picos cromatográficos do extrato EA. As características espectrais foram relacionadas a compostos fenólicos e flavonoídicos.

  20. Application of response surface methodology for air assisted-dispersive liquid- liquid microextraction of deoxynivalenol in rice samples prior to HPLC-DAD analysis and comparison with solid phase extraction cleanup.

    Science.gov (United States)

    Rahmani, Mashaallah; Ghasemi, Elham; Sasani, Mojtaba

    2017-04-01

    A fast, simple, and easy to operate air assisted-dispersive liquid-liquid microextraction (AA-DLLME) for preconcentration and extraction of deoxynivalenol (DON) from rice samples is proposed and compared with solid phase extraction (SPE) cleanup. DON was determined using a high performance liquid chromatography with diode-array detection (HPLC-DAD). AA-DLLME was performed using a glass syringe and dispersion of extractant in the sample solution was achieved with help of air bubbles. Chloroform was used as the extractant solvent. To find out the optimized condition for the proposed method, response surface methodology (RSM) was applied for multivariate optimization of effecting parameters namely volume of extractant, number of extraction, pH, and rate of centrifugation. Under optimized condition the dynamic range of calibration graph was found to be 50-500μgL(-1) with detection limit of 23.6μgL(-1). Both methods were applied for extraction of DON from rice.

  1. HPLC-DAD-q-TOF-MS as a powerful platform for the determination of phenolic and other polar compounds in the edible part of mango and its by-products (peel, seed, and seed husk).

    Science.gov (United States)

    Gómez-Caravaca, Ana María; López-Cobo, Ana; Verardo, Vito; Segura-Carretero, Antonio; Fernández-Gutiérrez, Alberto

    2016-04-01

    Free and bound phenolic and other polar compounds in mango edible fraction and its by-products (peel, seed, and seed husk) have been determined by HPLC-DAD-ESI-qTOF-MS. This analytical technique has demonstrated to be a valuable platform for the identification and quantification of these compounds in mango. In fact, UV-Vis and mass spectra data allowed the determination of 91 free compounds and 13 bound (cell wall linked) compounds taking into account the four fractions of mango. To our knowledge, this is the first time that mango seed husk has been studied regarding its phenolic compounds. The method proposed showed LODs between 0.006 and 0.85 μg/mL and accuracy ranged from 94.8 and 100.7%. Mango peel presented the highest concentration of free polar compounds followed by seed, pulp, and seed husk. It is also important to highlight that bound phenolic compounds had never been determined in mango pulp, seed, and seed husk before. Furthermore, ellagic acid was the most abundant bound compound in the four mango fractions analyzed. These results show that mango pulp and its by-products are a good source of phenolic and other polar compounds. In particular, mango seed contains a high total concentration of ellagic acid (650 mg/100 g dry weight).

  2. Antioxidant capacity, polyphenolic content and tandem HPLC-DAD-ESI/MS profiling of phenolic compounds from the South American berries Luma apiculata and L. chequén.

    Science.gov (United States)

    Simirgiotis, Mario J; Bórquez, Jorge; Schmeda-Hirschmann, Guillermo

    2013-08-15

    Native Myrtaceae fruits were gathered by South American Amerindians as a food source. At present, there is still some regional consume of the small berries from trees belonging to genus Luma that occurs in southern Chile and Argentina. The aerial parts and berries from Luma apiculata and Luma chequen were investigated for phenolic constituents and antioxidant capacity. A high performance electrospray ionisation mass spectrometry method was developed for the rapid identification of phenolics in polar extracts from both species. Thirty-one phenolic compounds were detected and 27 were identified or tentatively characterised based on photodiode array UV-vis spectra (DAD), ESI-MS-MS spectrometric data and spiking experiments with authentic standards. Twelve phenolic compounds were detected in L. apiculata fruits and 12 in the aerial parts while L. chequen yielded 10 compounds in fruits and 16 in aerial parts, respectively. From the compounds occurring in both Luma species, seven were identified as tannins or their monomers, 15 were flavonol derivatives and five were anthocyanins. The whole berry and aerial parts extracts presented high antioxidant capacity in the DPPH assay (IC50 of 10.41±0.02 and 2.44±0.03 μg/mL for L. apiculata, 12.89±0.05 and 3.22±0.05 for L. chequen, respectively), which can be related to the diverse range of phenolics detected. The antioxidant capacity together with the high polyphenolic contents and compounds identified can support at least in part, their use as botanical drugs. From the compounds identified in both species, 3-O-(6″-O-galloyl)-hexose derivatives of myricetin, quercetin, laricitrin and isorhamnetin are reported for the first time for the genus Luma.

  3. Rapid Screening for Flavone C-Glycosides in the Leaves of Different Species of Bamboo and Simultaneous Quantitation of Four Marker Compounds by HPLC-UV/DAD

    Directory of Open Access Journals (Sweden)

    Jin Wang

    2012-01-01

    Full Text Available A strategy for analyzing flavone C-glucosides in the leaves of different species of bamboo was developed. Firstly, the flavone C-glycosides were extracted from the bamboo leaves (51 species in 17 genera with methanol and chromatographed on silica gel 60 plates in automatic developing chamber (ADC2, and a qualitative survey using simple derivatization steps with the NP reagent was carried out. The flavone C-glycosides were found in 40 of 51 species of bamboo examined. Secondly, an HPLC method with photodiode array and multiple wavelength detector was optimized and validated for the simultaneous determination of flavone C-glycosides, including isoorientin, isovitexin, orientin, and vitexin in the leaves of three species of bamboo and the flavone C-glycosides were confirmed by LC/MS. The optimized HPLC method proved to be linear in the concentration range tested (0.2–100 μg/mL, r2≥0.9997, precise (RSD≤1.56%, and accurate (88–106%. The concentration ranges of isoorientin, isovitexin, orientin, and vitexin in three bamboo leaves samples were 1.00–2.78, 0–0.31, 0–0.07, and 0.20–0.68 mg/g, respectively. The proposed method was validated to be simple and reliable and can be a tool for quality control of bamboo leaf extract or its commercial products.

  4. Rapid Screening for Flavone C-Glycosides in the Leaves of Different Species of Bamboo and Simultaneous Quantitation of Four Marker Compounds by HPLC-UV/DAD.

    Science.gov (United States)

    Wang, Jin; Yue, Yong-de; Jiang, Hao; Tang, Feng

    2012-01-01

    A strategy for analyzing flavone C-glucosides in the leaves of different species of bamboo was developed. Firstly, the flavone C-glycosides were extracted from the bamboo leaves (51 species in 17 genera) with methanol and chromatographed on silica gel 60 plates in automatic developing chamber (ADC2), and a qualitative survey using simple derivatization steps with the NP reagent was carried out. The flavone C-glycosides were found in 40 of 51 species of bamboo examined. Secondly, an HPLC method with photodiode array and multiple wavelength detector was optimized and validated for the simultaneous determination of flavone C-glycosides, including isoorientin, isovitexin, orientin, and vitexin in the leaves of three species of bamboo and the flavone C-glycosides were confirmed by LC/MS. The optimized HPLC method proved to be linear in the concentration range tested (0.2-100 μg/mL, r(2) ≥ 0.9997), precise (RSD ≤ 1.56%), and accurate (88-106%). The concentration ranges of isoorientin, isovitexin, orientin, and vitexin in three bamboo leaves samples were 1.00-2.78, 0-0.31, 0-0.07, and 0.20-0.68 mg/g, respectively. The proposed method was validated to be simple and reliable and can be a tool for quality control of bamboo leaf extract or its commercial products.

  5. Analysis of flavone C-glycosides in the leaves of Clinacanthus nutans (Burm. f.) Lindau by HPTLC and HPLC-UV/DAD.

    Science.gov (United States)

    Chelyn, June Lee; Omar, Maizatul Hasyima; Mohd Yousof, Nor Syaidatul Akmal; Ranggasamy, Ramesh; Wasiman, Mohd Isa; Ismail, Zakiah

    2014-01-01

    Clinacanthus nutans (family Acanthaceae) has been used for the treatment of inflammation and herpes viral infection. Currently, there has not been any report on the qualitative and quantitative determination of the chemical markers in the leaves of C. nutans. The C-glycosidic flavones such as shaftoside, isoorientin, orientin, isovitexin, and vitexin have been found to be major flavonoids in the leaves of this plant. Therefore, we had developed a two-step method using thin-layer chromatography (TLC) and high pressure liquid chromatography (HPLC) for the rapid identification and quantification of the flavones C-glycosides in C. nutans leaves. The TLC separation of the chemical markers was achieved on silica gel 60 plate using ethyl acetate : formic acid : acetic acid : water (100 : 11 : 11 : 27 v/v/v/v) as the mobile phase. HPLC method was optimized and validated for the quantification of shaftoside, orientin, isovitexin, and vitexin and was shown to be linear in concentration range tested (0.4-200 μg/mL, r(2) ≥ 0.996), precise (RSD ≤ 4.54%), and accurate (95-105%). The concentration of shaftoside, orientin, vitexin, and isovitexin in C. nutans leave samples was 2.55-17.43, 0.00-0.86, 0.00-2.01, and 0.00-0.91 mmol/g, respectively.

  6. Chemical characteristics combined with bioactivity for comprehensive evaluation of Panax ginseng C.A. Meyer in different ages and seasons based on HPLC-DAD and chemometric methods.

    Science.gov (United States)

    Shan, Si-Ming; Luo, Jian-Guang; Huang, Fang; Kong, Ling-Yi

    2014-02-01

    Panax ginseng C.A. Meyer has been known as a valuable traditional Chinese medicines for thousands years of history. Ginsenosides, the main active constituents, exhibit prominent immunoregulation effect. The present study first describes a holistic method based on chemical characteristic and lymphocyte proliferative capacity to evaluate systematically the quality of P. ginseng in thirty samples from different seasons during 2-6 years. The HPLC fingerprints were evaluated using principle component analysis (PCA) and hierarchical clustering analysis (HCA). The spectrum-efficacy model between HPLC fingerprints and T-lymphocyte proliferative activities was investigated by principal component regression (PCR) and partial least squares (PLS). The results indicated that the growth of the ginsenosides could be grouped into three periods and from August of the fifth year, P. ginseng appeared significant lymphocyte proliferative capacity. Close correlation existed between the spectrum-efficacy relationship and ginsenosides Rb1, Ro, Rc, Rb2 and Re were the main contributive components to the lymphocyte proliferative capacity. This comprehensive strategy, providing reliable and adequate scientific evidence, could be applied to other TCMs to ameliorate their quality control.

  7. HPLC-DAD-ELSD法同时测定黄芪中5个成分的含量%Simultaneous determination of five compounds in Radix Astragali by high-performance liquid chromatography coupled with diode arry and evaporative light scattering detectors

    Institute of Scientific and Technical Information of China (English)

    梁瑾; 刘小花; 任远; 封士兰

    2013-01-01

    目的:建立HPLC-DAD-ELSD法同时测定黄芪药材中黄酮类和皂苷类成分的方法,比较15批不同产地的黄芪药材中各有效成分的含量.方法:采用HPLC-DAD-ELSD联用法,乙腈和水不同比例梯度洗脱,流速1 mL·min-1,检测波长254nm;蒸发光散射检测器(ELSD)漂移管温度112.8℃,载气流速3.2 L·min-1,同时测定黄芪药材中毛蕊异黄酮、芒柄花素、黄芪甲苷(黄芪皂苷Ⅳ)、黄芪皂苷Ⅱ和黄芪皂苷Ⅲ的含量.结果:通过1次进样,同时测定了2个黄酮类和3个皂苷类等成分的含量,且线性关系良好(r =0.9992~0.9999).平均回收率为99.1%~100.9%.结论:该方法简便,重复性好,可用于黄芪药材中黄酮类和皂苷类成分的同时测定,也可用于黄芪药材的质量控制.%Objective: To establish a quantitative method of two major active isoflavonids and three main saponins in Radix Astragali by high - performance liquid chromatography coupled with diode any and evaporative light scattering detectors ( HPLC - DAD - ELSD ) , and compare the contents of calycosin, formononetin and astragaloside Ⅳ,Ⅱ,Ⅲ among 15 batches from different habitats. Methods:HPLC - DAD - ELSD was conducted on a Spursil(TM)C18 column(5 μm,250 mm ×4. 6 mm)with gradient elution ,the mobile phase was ACN and H2O,the temperature of drift tube was 112. 8 ℃ and the nebulizing gas flow rate was 3. 2 L · min-1. Results: Five compounds were simultaneously detected with one injection, good linear relationships were found ( r = 0. 9992 - 0. 9999 ) , and the average recovery rates were 99. 13% - 100. 94%. Conclusions:The HPLC - DAD - ELSD method is simple and reproducible, which can be used as principles of quantitative analysis.

  8. Determination of memantine in plasma and vitreous humour by HPLC with precolumn derivatization and fluorescence detection.

    Science.gov (United States)

    Puente, Belen; Hernandez, Esther; Perez, Susana; Pablo, Luis; Prieto, Esther; Garcia, Maria Angeles; Bregante, Miguel Angel

    2011-01-01

    A new HPLC procedure with precolumn derivatization and rimantadine as the internal standard for determining memantine, a candidate agent for the treatment of glaucoma in plasma and vitreous humour, has been developed and validated. Precolumn derivatization was performed with 9-fluorenylmethyl-chloroformate-chloride (FMOC-Cl) as the derivatization reagent and followed by a liquid-liquid extraction with n-hexane. Optimal conditions for derivatization were an FMOC-Cl concentration of 1.5 mM, a reaction time of 20 min, the temperature at 30°C, the borate buffer pH 8.5, and a borate buffer-acetonitrile ratio of 1:1. The derivatives were analyzed by isocratic HPLC with the fluorescence detector λex 260 nm λem 315 nm on a Novapack C(18) reversed-phase column with a mobile phase of acetonitrile-water (73:27, v/v), 40°C, and a flow rate of 1.2 mL/min. The linear range was 10-1000 ng/mL with a quantification limit of ∼ 10 ng/mL for both types of samples. This analytical method may be suitable for using in ocular availability studies.

  9. Thiamin analysis in red wine by fluorescence reverse phase-HPLC.

    Science.gov (United States)

    Liddicoat, Callum; Hucker, Barry; Liang, Hao; Vriesekoop, Frank

    2015-06-15

    The derivatization of thiamin vitamers to their respective thiochrome by ferricyanide to facilitate fluorescence detection following separation by HPLC provides a powerful analytical tool. However the polyphenolic compounds in red wine readily interact with ferricyanide, reducing the effectiveness of ferricyanide oxidation in the derivatization of thiamin. We describe a method to facilitate the removal of polyphenolic compounds that interfere with the ferricyanide derivatization of thiamin. Polyvinylpolypyrrolidone afforded the total removal of phenolic compounds from red wines and allowed a spike recovery of thiamin vitamers (101% for thiamin; 104% for TMP; and 100% for TDP) in a wide range of red wines. This research found that Merlot styles of red wine contained the highest concentration of total thiamin (29.01 ng/mL) while Pinot Noir wines contained the lowest total concentration (8.27 ng/mL). Copyright © 2015 Elsevier Ltd. All rights reserved.

  10. Simultaneous Determination of Amlodipine Besylate and Atorvastatin Calcium in Binary Mixture by Spectrofluorimetry and HPLC Coupled with Fluorescence Detection

    OpenAIRE

    Bahia A. Moussa; Asmaa A. El-Zaher; Mahrouse, Marianne A.; Ahmed, Maha S.

    2013-01-01

    Caduet tablets are novel prescription drug that combines amlodipine besylate (AM) with atorvastatin calcium (AT). A spectrofluorimetric and an HPLC-fluorescence detection methods were developed for simultaneous determination of both drugs in tablets. In the spectrofluorimetric method, native fluorescence of AM and AT were measured in methanol at 442 and 369 nm upon excitation at 361 and 274 nm, respectively. The emission spectrum of each drug reveals zero value at the emission wavelength of t...

  11. Quantitation of chlorophylls and 22 of their colored degradation products in culinary aromatic herbs by HPLC-DAD-MS and correlation with color changes during the dehydration process.

    Science.gov (United States)

    Lafeuille, Jean-Louis; Lefèvre, Stéphane; Lebuhotel, Julie

    2014-02-26

    Chlorophylls and their green and olive-brown derivatives were successfully separated from culinary herb extracts by HPLC with photodiode-array and mass spectrometry detection. The method involved a ternary gradient elution and reverse-phase separation conditions capable of resolving 24 different pigments (2 chlorophylls and 22 of their derivatives) of different polarities within 28 min. The method was applied to monitor color changes in 50 samples of culinary aromatic herbs subjected to five different drying treatments. Of the 24 pigments, 14 were key to understanding the differences between the primary degradation pathways of chlorophyll a and chlorophyll b in culinary herbs during drying processes. A color degradation ladder based on the total molar percentage of all the remaining green pigments was also proposed as a tool to measure the impact of drying treatments on aromatic herb visual aspects.

  12. Characteristic fingerprint based on gingerol derivative analysis for discrimination of ginger (Zingiber officinale) according to geographical origin using HPLC-DAD combined with chemometrics.

    Science.gov (United States)

    Yudthavorasit, Soparat; Wongravee, Kanet; Leepipatpiboon, Natchanun

    2014-09-01

    Chromatographic fingerprints of gingers from five different ginger-producing countries (China, India, Malaysia, Thailand and Vietnam) were newly established to discriminate the origin of ginger. The pungent bioactive principles of ginger, gingerols and six other gingerol-related compounds were determined and identified. Their variations in HPLC profiles create the characteristic pattern of each origin by employing similarity analysis, hierarchical cluster analysis (HCA), principal component analysis (PCA) and linear discriminant analysis (LDA). As results, the ginger profiles tended to be grouped and separated on the basis of the geographical closeness of the countries of origin. An effective mathematical model with high predictive ability was obtained and chemical markers for each origin were also identified as the characteristic active compounds to differentiate the ginger origin. The proposed method is useful for quality control of ginger in case of origin labelling and to assess food authenticity issues.

  13. Analysis of Flavone C-Glycosides in the Leaves of Clinacanthus nutans (Burm. f. Lindau by HPTLC and HPLC-UV/DAD

    Directory of Open Access Journals (Sweden)

    June Lee Chelyn

    2014-01-01

    Full Text Available Clinacanthus nutans (family Acanthaceae has been used for the treatment of inflammation and herpes viral infection. Currently, there has not been any report on the qualitative and quantitative determination of the chemical markers in the leaves of C. nutans. The C-glycosidic flavones such as shaftoside, isoorientin, orientin, isovitexin, and vitexin have been found to be major flavonoids in the leaves of this plant. Therefore, we had developed a two-step method using thin-layer chromatography (TLC and high pressure liquid chromatography (HPLC for the rapid identification and quantification of the flavones C-glycosides in C. nutans leaves. The TLC separation of the chemical markers was achieved on silica gel 60 plate using ethyl acetate : formic acid : acetic acid : water (100 : 11 : 11 : 27 v/v/v/v as the mobile phase. HPLC method was optimized and validated for the quantification of shaftoside, orientin, isovitexin, and vitexin and was shown to be linear in concentration range tested (0.4–200 μg/mL, r2 ≥ 0.996, precise (RSD ≤ 4.54%, and accurate (95–105%. The concentration of shaftoside, orientin, vitexin, and isovitexin in C. nutans leave samples was 2.55–17.43, 0.00–0.86, 0.00–2.01, and 0.00–0.91 mmol/g, respectively.

  14. Multiresidue determination of acidic pesticides in water by HPLC-DAD with confirmation by GC-MS using conversion to the methyl ester with trimethylsilyldiazomethane.

    Science.gov (United States)

    Moy, Thomas W; Brumley, William C

    2003-08-01

    A multiresidue pesticide methodology is studied and results for acidics are reported here, with base/neutral to follow. This work studies a literature procedure as a possible general approach to many pesticides and potentially other analytes that are considered to be liquid chromatographic (LC) candidates, rather than gas chromatographic (GC) ones. The analysis of the sewage effluent of a major southwestern U.S. city serves as an example of the application of the methodology to a real sample. Recovery studies were also conducted to validate the proposed extraction step. A gradient elution program was followed for the high-performance LC (HPLC) leading to a general approach for acidics. Confirmation of identity was by electron ionization-GC-mass spectrometry (MS) after conversion of the acids to the methyl ester (or other appropriate methylation) by means of trimethylsilyldiazomethane. The 3,4-dichlorophenoxyacetic acid is used as an internal standard to monitor the reaction, and polychlorobiphenyl #19 is used for the quantitation internal standard. Although others have reported similar analyses of acids, conversion to the methyl ester is by means of diazomethane itself, rather than by the more convenient and safe trimethylsilyldiazomethane. Thus, the present paper supports the use of trimethylsilyldiazomethane with all of these acids (trimethylsilyldiazomethane has been used in environmental work with some phenoxyacetic acid herbicides) and further supports the usefulness of this reagent as a potential replacement for diazomethane. The HPLC approach here could also serve as the separation basis for an LC-MS solution for confirmation of identity, as well as quantitation.

  15. A new estimation of the total flavonoids in silkworm cocoon sericin layer through aglycone determination by hydrolysis-assisted extraction and HPLC-DAD analysis

    Directory of Open Access Journals (Sweden)

    Jin-Ge Zhao

    2016-03-01

    Full Text Available Background: Silk sericin and a few non-protein components isolated from the cocoon layer including two silk proteins in silkworm Bombyx mori has many bioactivities. The dietary sericin possess antinatural oxidation, anticancer, antihyperlipidemic, and antidiabetic activities. The non-protein components surrounding the sericin layer involve in wax, pigments mainly meaning flavonoids, sugars, and other impurities. However, very few investigations have reported the estimation of the total flavonoids derived from the cocoon layer. The flavonoids are commonly present in their glycosylated forms and mostly exist as quercetin glycosides in the sericin layers of silkworm cocoons. Objective: The aim of this study was to find a more accurate method to estimate the level of the total flavonoids in silkworm cocoons. Design: An efficient procedure of hydrolysis-assisted extraction (HAE was first established to estimate the level of the total flavonoids through the determination of their aglycones, quercetin, and kaempferol. Then, a comparison was made between traditional colorimetric method and our method. In addition, the antioxidant activities of hydrolysis-assisted extract sample were determined. Results: The average contents of quercetin and kaempferol were 1.98 and 0.42 mg/g in Daizo cocoon. Their recoveries were 99.56 and 99.17%. The total sum of quercetin and kaempferol was detected to be 2.40±0.07 mg/g by HAE-HPLC, while the total flavonoids (2.59±0.48 mg/g estimated by the traditional colorimetric method were only equivalent to 1.28±0.04 mg/g of quercetin. The HAE sample also exhibits that IC50 values of scavenging ability of diphenyl picryl hydrazinyl (DPPH radical and hydroxyl radical (HO· are 243.63 µg/mL and 4.89 mg/mL, respectively. Conclusions: These results show that the HAE-HPLC method is specificity of cocoon and far superior to the colorimetric method. Therefore, this study has profound significance for the comprehensive utilization

  16. HPLC-DAD和UPLC-MS/MS对蔓花生中二苯乙烯类化合物的研究%Simultaneous Determination of Stilbene Compounds in Arachis duranensis by HPLC-DAD and UPLC-MS/MS

    Institute of Scientific and Technical Information of China (English)

    韩寒冰; 王明阳; 马超; 李海航

    2014-01-01

    采用HPLC-DAD和UPLC-MS/MS对蔓花生中白藜芦醇、白藜芦醇苷和二苯乙烯苷三种二苯乙烯类化合物进行鉴定,建立HPLC-DAD同时测定三种化合物含量的方法.HPLC-DAD分析采用依利特ODS1色谱柱,柱温40℃,流动相A为甲醇,B为1%乙酸水溶液,流速1.0 mL/min.UPLC-MS/MS采用ACQUITY UPLC BEH C18色谱柱,流动相A为乙腈,B为水,电喷雾离子源,负离子检测,数据采集方式为多反应监测模式(MRM).结果表明:蔓花生根、茎和叶中白藜芦醇含量分别为66.28、90.83和81.56 μg/g,白藜芦醇苷含量分别为123.78,302.77和236.53 μg/g,根和茎中二苯乙烯苷含量分别为673.60和764.65 μg/g,叶中未检测到二苯乙烯苷.

  17. Validation of a multi-analyte HPLC-DAD method for determination of uric acid, creatinine, homovanillic acid, niacinamide, hippuric acid, indole-3-acetic acid and 2-methylhippuric acid in human urine.

    Science.gov (United States)

    Remane, Daniela; Grunwald, Soeren; Hoeke, Henrike; Mueller, Andrea; Roeder, Stefan; von Bergen, Martin; Wissenbach, Dirk K

    2015-08-15

    During the last decades exposure sciences and epidemiological studies attracts more attention to unravel the mechanisms for the development of chronic diseases. According to this an existing HPLC-DAD method for determination of creatinine in urine samples was expended for seven analytes and validated. Creatinine, uric acid, homovanillic acid, niacinamide, hippuric acid, indole-3-acetic acid, and 2-methylhippuric acid were separated by gradient elution (formate buffer/methanol) using an Eclipse Plus C18 Rapid Resolution column (4.6mm×100mm). No interfering signals were detected in mobile phase. After injection of blank urine samples signals for the endogenous compounds but no interferences were detected. All analytes were linear in the selected calibration range and a non weighted calibration model was chosen. Bias, intra-day and inter-day precision for all analytes were below 20% for quality control (QC) low and below 10% for QC medium and high. The limits of quantification in mobile phase were in line with reported reference values but had to be adjusted in urine for homovanillic acid (45mg/L), niacinamide 58.5(mg/L), and indole-3-acetic acid (63mg/L). Comparison of creatinine data obtained by the existing method with those of the developed method showing differences from -120mg/L to +110mg/L with a mean of differences of 29.0mg/L for 50 authentic urine samples. Analyzing 50 authentic urine samples, uric acid, creatinine, hippuric acid, and 2-methylhippuric acid were detected in (nearly) all samples. However, homovanillic acid was detected in 40%, niacinamide in 4% and indole-3-acetic acid was never detected within the selected samples.

  18. Enantioselective HPLC-DAD method for the determination of etodolac enantiomers in tablets, human plasma and application to comparative pharmacokinetic study of both enantiomers after a single oral dose to twelve healthy volunteers.

    Science.gov (United States)

    Hewala, Ismail I; Moneeb, Marwa S; Elmongy, Hatem A; Wahbi, Abdel-Aziz M

    2014-12-01

    An enantioselective high performance liquid chromatographic method with diode array detection (HPLC-DAD) was developed and validated for the determination of etodolac enantiomers in tablets and human plasma. Enantiomeric separation was achieved on a Kromasil Cellucoat chiral column (250 mm × 4.6mm i.d., 5 µm particle size) using a mobile phase consisting of hexane: isopropanol: triflouroacetic acid (90:10:0.1 v/v/v) at a flow rate of 1.0 mL min(-1). The chromatographic system enables the separation of the two enantiomers and the internal standard within a cycle time of 8 min. The resolution between the two enantiomers was 4.25 and the resolution between each enantiomer and the internal standard was more than 2.0. Detection was carried out at 274 nm, and the purity assessment was performed using a photodiode array detector. Solid phase extraction technique using C-18 cartridge was applied to extract the analytes from the plasma samples, and the percentage recovery was more than 95% for the lower quantification limit. The method has been validated with respect to selectivity, linearity, accuracy and precision, robustness, limit of detection and limit of quantification. The validation acceptance criteria were met in all cases. The linearity range for the determination of each enantiomer in human plasma was 0.4-30.0 µg mL(-1) and the limits of quantification of R-etodolac and S-etodolac were 0.20 and 0.19 µg mL(-1), respectively. The validated method was successfully applied to the determination of etodolac enantiomers in tablets and to a comparative pharmacokinetic study of the two enantiomers after the administration of 300 mg single oral dose etodolac racemate tablets to twelve healthy volunteers.

  19. Two Ganoderma species: profiling of phenolic compounds by HPLC-DAD, antioxidant, antimicrobial and inhibitory activities on key enzymes linked to diabetes mellitus, Alzheimer's disease and skin disorders.

    Science.gov (United States)

    Zengin, Gokhan; Sarikurkcu, Cengiz; Gunes, Erdogan; Uysal, Ahmet; Ceylan, Ramazan; Uysal, Sengul; Gungor, Halil; Aktumsek, Abdurrahman

    2015-08-01

    This work reports the antioxidant, antimicrobial, and inhibitory effects of methanol and water extracts from Ganoderma applanatum (GAM: methanol extract and GAW: water extract) and G. resinaceum (GRM: methanol extract and GRW: water extract) against cholinesterase, tyrosinase, α-amylase and α-glucosidase. The total phenolics, flavonoids contents, and HPLC profile of phenolic components present in the extracts, were also determined. Antioxidant activities were investigated by using different assays, including DPPH, ABTS, FRAP, CUPRAC, phosphomolybdenum and metal chelating assays. Antimicrobial activity of the tested Ganoderma extracts was also studied by the broth microdilution method. Generally, the highest antioxidant (59.24 mg TEs per g extract for DPPH, 41.32 mg TEs per g extract for ABTS, 41.35 mg TEs per g extract for CUPRAC, 49.68 mg TEs per g extract for FRAP, 130.57 mg AAEs per g extract for phosphomolybdenum and 26.92 mg EDTAEs per g extract) and enzyme inhibitory effects (1.47 mg GALAEs per g extract for AChE, 1.51 mg GALAEs per g extract for BChE, 13.40 mg KAEs per g extract for tyrosinase, 1.13 mmol ACEs per g extract for α-amylase and 2.20 mmol ACEs per g extract for α-glucosidase) were observed in GRM, which had the highest concentrations of phenolics (37.32 mg GAEs g(-1) extract). Again, Ganoderma extracts possess weak antibacterial and antifungal activities. Apigenin and protocatechuic acid were determined as the main components in GRM (1761 μg per g extract) and GAM (165 μg per g extract), respectively. The results suggest that the Ganoderma species may be considered as a candidate for preparing new food supplements and can represent a good model for the development of new drug formulations.

  20. A novel validated RP-HPLC-DAD method for the simultaneous estimation of Metformin Hydrochloride and Canagliflozin in bulk and pharmaceutical tablet dosage form with forced degradation studies

    Directory of Open Access Journals (Sweden)

    Uttam Prasad Panigrahy

    2015-09-01

    Full Text Available A novel approach was used to develop and validate a rapid isocratic Reversed Phase-High Performance Liquid Chromatographic method for the simultaneous estimation of Metformin Hydrochloride and Canagliflozin in bulk and pharmaceutical tablet dosage form with forced degradation studies. The separation was performed by using Kromasil C18 column (250mm×4.6 mm, 5mm particle size, Waters Alliance e2695 HPLC system with 2998 PDA detector and mobile phase contained a mixture of 0.01M Ammonium acetate (pH adjusted to 3.5 with orthophosphoric acid and Acetonitrile (65:35, v/v. The flow rate was set to 1ml/min with responses measured at 254nm. The retention time of Metformin Hydrochloride and Canagliflozin was 2.440min and 3.713min respectively with resolution of 8.95.Linearity was established in the range of 50-300µg/ml for Metformin Hydrochloride and 5-30µg/ml for Canagliflozin with correlation coefficients (r2=0.999. The percentage recoveries were between (99.45%-100.65% and (99.95%-100.74% for Metformin Hydrochloride and Canagliflozin respectively. Validation parameters were evaluated according to the International Conference on Harmonization (ICH Q2 R1 guidelines. The forced degradation studies were performed by using HCl, NaOH, H2O2, thermal, UV radiation and water. Metformin Hydrochloride and Canagliflozin are more sensitive towards oxidative degradation condition. The developed method was successfully applied for the quantification and hyphenated instrumental analysis.

  1. Determination of ethinylestradiol and levonorgestrel in oral contraceptives with HPLC methods with UV detection and UV/fluorescence detection

    Directory of Open Access Journals (Sweden)

    Zorica Arsova-Sarafinovska

    2006-06-01

    Full Text Available Oral contraceptives are pharmaceutical formulations containing an estrogen in a small amount and a synthetic progestin in 5-30 times bigger amount. A sensitive, accurate and rapid method for determination of active compounds is required. We have developed HPLC methods for determination of ethinylestradiol (EED and levonorgestrel (LNG in commercially available tablets. Chromatographic separation was performed on a Purospher® STAR RP-18e reversed-phase column (150 X 4.0 mm I.D.; particle size 5 µm in an isocratic mode with a mobile phase constituted of 47% acetonitrile: 53% water (V/V for both methods. The elution was carried out at a flow rate of 1.50 ml /min. All analyses were performed at room temperature (24 +/- 2°C. In the HPLC method with UV detection (internal standard method both compounds were detected at 215 nm. Drospirenone was used as an internal standard. In HPLC method with UV/fluorescence detection (external standard method LNG was monitored at 242 nm, while EED was detected with fluorescence detector at 310 nm (excitation 285 nm. The methods’ performances were fully validated by a determination of linearity, reproducibility, accuracy and sensitivity. Both methods were applied for determination of Uniformity of Dosage Units. The results obtained with both methods were highly comparable. However, the HPLC method with UV/ fluorescence detection has showed superior sensitivity for EED indicated by 83 times lower detection limit. HPLC method with UV/ fluorescence detection could be recommended as a method of choice for determination of ethinylestradiol, present at a very low dosage level in low-dose oral contraceptives, that also contain bigger amount of synthetic progestin.

  2. Sensitive determination of ranitidine in rabbit plasma by HPLC with fluorescence detection.

    Science.gov (United States)

    Khedr, Alaa

    2008-02-01

    A sensitive high-performance liquid chromatographic method for determination of ranitidine (RAN) in rabbit plasma is described. The method is based on liquid-liquid extraction, labeling with dansyl chloride and monitoring with fluorescence detector at 338nm (ex)/523nm (em). Plasma samples were extracted with diethyl ether alkalinized with 1M sodium hydroxide. Ephedrine HCl (EPH-HCl) was used as internal standard. Both, RAN and EPH were completely derivatized after heating at 60 degrees C for 10min in sodium bicarbonate solution (pH 9.5). The derivatized samples were analyzed by HPLC using Agilent Zorbax Extended C18 column (150mmx4.6mm i.d.) and mobile phase consists of 48% acetonitrile and 52% sodium acetate solution (0.02M, pH 4.6). The linearity of the method was in the range of 0.025-10microg/ml. The limits of detection (LOD) and quantification (LOQ) were 7.5+/-0.18 and 22.5+/-0.12ng/ml, respectively. Ranitidine recovery was 97.5+/-1.1% (n=6; R.S.D.=1.8%). The method was applied on plasma collected from rabbits at different time intervals after oral administration of 5mg/kg ranitidine HCl.

  3. A new total antioxidant potential measurements using RP-HPLC assay with fluorescence detection.

    Science.gov (United States)

    Głód, Bronisław K; Piszcz, Paweł; Czajka, Katarzyna; Zarzycki, Paweł K

    2011-05-01

    In this paper, an improved total antioxidant potential (TAP) estimation using high-performance liquid chromatographic (HPLC) assay with fluorometric detection has been described. The principle of this method is based on the hydroxyl radicals generated in the Fenton-like reaction and subsequently detected using hydroxyterephthalic acid (HTPA), which is a reaction product of hydroxyl radicals and terephthalic acid (TPA), working as a sensing compound. HTPA quantity in the samples was measured by fluorescence detector working at excitation and emission wavelengths equal to 312 and 428 nm, respectively. A number of key experimental conditions including the influence of the reaction (incubation) time on the surface areas of HTPA peaks, concentration of Fe(II) ions as well as the influence of concentration of TPA on the surface area of the chromatographic peak of HTPA were optimized to the characteristic feature of TAP measurements. The elaborated assay has been used to evaluate TAP values of selected low-molecular mass compounds like pyrogallol, tryptamine, and n-alcohols (methanol, ethanol, and n-propanol) as well as chlorogenic and ascorbic acids and benzoic acid derivatives, which are commonly present in the food samples.

  4. 蚊香中有效成分和仲丁威含量的HPLC-DAD测定%Determination of the active ingredients and fenobucarb content in mosquito-repellent incense by HPLC-DAD

    Institute of Scientific and Technical Information of China (English)

    武中平; 高巍; 俞幼芬

    2008-01-01

    采用HPLC-DAD法同时测定了蚊香中有效成分和仲丁威的含量.样品经乙腈-丙酮提取后,用二极管阵列检测器、C18 柱,以乙腈+水为流动相,对蚊香中有效成分和仲丁威的同时测定可取得满意的结果.该方法的相对标准偏差为0.13%~0.27%,样品平均加标回收率为 98.34%~102.03%.

  5. Determination of two flavonoids in Tongxian grapefruit leaves by HPLC-DAD%HPLC-DAD法测定通贤柚叶中两种活性黄酮的含量

    Institute of Scientific and Technical Information of China (English)

    黄友秋; 赵雪荔; 陈席; 游元元

    2015-01-01

    目的:建立HPLC-DAD测定通贤柚叶中柚皮苷与野漆树苷的方法.方法:色谱柱为Sepax Amethyst C18-H(250×4.60mm,5μm),流动相为乙腈-0.1%甲酸梯度洗脱,流速1.0mL/min,检测波长为283nm(柚皮苷),337nm(野漆树苷),柱温25℃,选样量20μL.结果:柚皮苷与野漆树苷色谱峰分离良好.二者分别在0.0653~0.6526mg/mL与0.0350~0.3505mg/mL范围内与峰面积呈良好的线性关系.平均加样回收率分别为98.68%与98.47%.结论:所建立的含量测定方法可用于通贤柚叶的质量控制.采收季节与叶龄对柚叶中两种黄酮类物质的含量有较明显的影响.

  6. Determination of Vitamin D3 and 25-Hydroxyvitamin D3 in Meat by HPLC UV-DAD and LC-MS/MS%HPLC-UV-DAD和LC-MS/MS法测定鲜肉中维生素D3和25-OH维生素D3

    Institute of Scientific and Technical Information of China (English)

    张洁; 张欣烨; 张伟; 张利锋; 马青青

    2016-01-01

    建立了HPLC-UV-DAD和LC-MS/MS测定肉中维生素D3和25-OH维生素D3的方法.样品经皂化、有机相萃取和SPE净化小柱后,维生素D3用反相高效液相色谱(HPLC)配有紫外二极管阵列检测器(UV-DAD)检测,维生素25-OH-D3用三重四级杆质谱(APCI-MS/MS)测定,定量采用维生素D2和25-OH维生素D2作为内标物质,有证的标准物质维生素D3和25-OH维生素D3作为质量控制工具.结果表明,2种物质的检出限分别是0.10μg/100 g和0.04 μg/100 g,方法线性范围10~200 ng/mL,相关系数达0.99以上,回收率>84.2%,方法的精密度范围为0.01%~4.19%.该方法适用于测定肉(猪肉、牛肉、鸡蛋和鱼等)中维生素D3和25-OH-维生素D3.

  7. Determination of the Rhein in Runchang Pills (Concentrated pill) by HPLC-DAD%HPLC-DAD测定润肠丸(浓缩丸)中大黄酸成分

    Institute of Scientific and Technical Information of China (English)

    朱仁愿; 蒲玉红

    2015-01-01

    目的建立润肠丸中大黄酸成分的高效液相色谱测定方法。方法:Phenomenex Luna C18柱(5um,150x4.60mm,100A);流动相:乙腈-0.1%磷酸(60:40);流速:1.0mL·min-1;二极管阵列检测器,检测波长:254nm;柱温:40℃。结果:该方法测定润肠丸中大黄酸成分时,线性关系良好,r2=1.000,平均回收率(n=6)为99.35%。结论:方法简便,快速准确,可作为样品的检测方法。%Objective To establish an HPLC method for the determination of the contents of rhein in Runchang pil s.Methods The separation was performed on Phenomenex Luna C18 column(5um, 150x4.60mm,100A)with a mobile phase of acetonitrile-1%Phosphoric acid(60:40),and the flow rate were 1.0mL·min-1.Photodiode Ar ay detector was used and the wavelength of detector for rhein was set at 254nm,and 40℃ for column.Results The method for determining runchang pil in rhein composition, linear relationship is good, r2 = 1.000, and the average recovery (n = 6) was 99.35%. Conclusion The method is accurate and sensitive.

  8. HPLC-DAD simultaneous determination of four vitamins in compound vitamin B tablets%HPLC-DAD法同时测定复合维生素B片中4种维生素的含量

    Institute of Scientific and Technical Information of China (English)

    陈金泉; 王军; 何立环

    2012-01-01

    Objective:To establish a method for determination of four vitamins in Compound vitamin B tables. Methods:RP-HPLC was adaptded on a column of ZORBAX Eclipse XDB-C18 (4. 6 mm ×250 mm ,5 μm), ga-dient elution with the mobile phase of A;0.05 mol o L-1 potassium dihydrogen phosphate(pH 4.6 adjusted with 0. 1% triethylamine,) and B:methanol at a flow rate of 1.0 mL o min-1 and the detection wavelength was 266 nm with the column temperatureat was 40 ℃. Results:In the linear range the correlation coefficients ranged from 0.9995-0. 9999. The average recoveries were 98.4% -100.4% with corresponding RSD of 0. 6% -2.0%.Con-clution:This method is simple, rapid, sensitive and accurate. It is available to the quality control of compound vitamin B tablets.%目的:建立反相高效液相色谱法同时测定复合维生素B片中4种维生素含量.方法:采用梯度洗脱反相高效液相色谱法,ZORBAX Ecliose XDB-C18(4.6mm×250mm,5μm)色谱柱,流动相A为含0.1%三乙胺的0.05mol·L-1磷酸二氢钾溶液(pH4.6);流动相B为甲醇,检测波长为266nm、柱温为40℃、流速为1.0 mL·min-1.结果:4种维生素含量测定方法的线性关系良好,r0.999 5~0.999 9,方法 回收率均在99.2~ 100.6%范围内,RSD均在0.8% ~2.0%之间.结论:本方法简便快捷,灵敏准确,可以用于复合维生素B片的质量控制.

  9. Quantitative determination of four constituents of Tinospora sps. by a reversed-phase HPLC-UV-DAD method. Broad-based studies revealing variation in content of four secondary metabolites in the plant from different eco-geographical regions of India.

    Science.gov (United States)

    Ahmed, S M; Manhas, L R; Verma, V; Khajuria, R K

    2006-09-01

    This paper describes the separation and quantitation of important markers, such as 20beta-hydroxyecdysone, tinosporaside, cordioside, and columbin, present in three species of Tinospora viz, T. cordifolia, T. malabrica, and T. crispa. A reverse-phase (RP) high-performance liquid chromatography (HPLC)-UV-diode array detection (DAD) method employing gradient elution is thus developed. The marker compounds isolated from 70% ethanolic extract of T. cordfolia by repeated column chromatography are identified on the basis of (1)H NMR, (13)C NMR, and mass spectral data. The compounds are separated on a RP (RP-18, 5 microm, 250 x 4.6-mm i.d.) column using water-acetonitrile gradient and are detected by the HPLC-UV-DAD method. The calibration curves that result from marker compounds in the concentration range of 100-2000 ng on column exhibit a good correlation (r(2) > or = 0.99978). The method is successfully applied to separate and study the content of four marker compounds in 40 different accessions of three Tinospora species collected from different regions of India. The studies reveal that the maximum amount of the marker compounds is present in Tinospora cordifolia species, especially from accessions collected from higher altitudes of the Jammu province (North India).

  10. Determination of roxithromycin in human plasma by HPLC with fluorescence and UV absorbance detection: application to a pharmacokinetic study.

    Science.gov (United States)

    Główka, Franciszek K; Karaźniewicz-Łada, Marta

    2007-06-01

    A selective HPLC method with fluorescence detection for the determination of roxithromycin (ROX) in human plasma was described. After solid-phase extraction (SPE), ROX and erythromycin (internal standard, I.S.) were derivatized by treatment with 9-fluorenylmethyl chloroformate (FMOC-Cl). Optimal resolution of fluorescence derivatives of ROX and I.S. was obtained during one analytical run using reversed phase, C(18) column. The mobile phase was composed of potassium dihydrogenphosphate solution, pH 7.5 and acetonitrile. Fluorescence of the compounds was measured at the maximum excitation, 255 nm and emission, 313 nm, of ROX derivatives. Validation parameters of the method were also established. After SPE, differences in recoveries of ROX and erythromycin from human plasma were observed. The linear range of the standard curve of ROX in plasma was 0.5-10.0 mg/l. The validated method was successfully applied for pharmacokinetic studies of ROX after administration of a single tablet of ROX.

  11. A sensitive and robust HPLC assay with fluorescence detection for the quantification of pomalidomide in human plasma for pharmacokinetic analyses.

    Science.gov (United States)

    Shahbazi, Shandiz; Peer, Cody J; Polizzotto, Mark N; Uldrick, Thomas S; Roth, Jeffrey; Wyvill, Kathleen M; Aleman, Karen; Zeldis, Jerome B; Yarchoan, Robert; Figg, William D

    2014-04-01

    Pomalidomide is a second generation IMiD (immunomodulatory agent) that has recently been granted approval by the Food and Drug Administration for treatment of relapsed multiple myeloma after prior treatment with two antimyeloma agents, including lenalidomide and bortezomib. A simple and robust HPLC assay with fluorescence detection for pomalidomide over the range of 1-500ng/mL has been developed for application to pharmacokinetic studies in ongoing clinical trials in various other malignancies. A liquid-liquid extraction from human plasma alone or pre-stabilized with 0.1% HCl was performed, using propyl paraben as the internal standard. From plasma either pre-stabilized with 0.1% HCl or not, the assay was shown to be selective, sensitive, accurate, precise, and have minimal matrix effects (HPLC-FL assay allows a broader range of laboratories to measure pomalidomide for application to clinical pharmacokinetics.

  12. HPLC-DAD法同时测定更年安片中6个活性成分的含量%HPLC-DAD simultaneous determination of six active ingredients in Gengnian' an tablets

    Institute of Scientific and Technical Information of China (English)

    施法; 侯峰; 陆军; 郭汉文; 张满来; 郝延军

    2012-01-01

    目的:建立同时分析测定更年安片中6个活性成分(2,3,5,4'-四羟基二苯乙烯-2 -O-β-D-葡萄糖苷、毛蕊花糖苷、仙茅苷、丹皮酚、哈巴俄苷、五味子醇甲)的方法.方法:采用HPLC-DAD法,色谱柱为奥泰Alltima ODS( 250 mm×4.6mm,5μm),以乙腈为流动相A,0.05%磷酸溶液为流动相B,梯度洗脱,流速1.0mL·min-1,检测波长为254 nm(测定五味子醇甲)、284 nm(测定仙茅苷、丹皮酚、哈巴俄苷)、330 nm(测定2,3,5,4'-四羟基二苯乙烯-2 -O-β-D-葡萄糖苷、毛蕊花糖苷).结果:2,3,5,4'-四羟基二苯乙烯-2-O-β-D-葡萄糖苷、毛蕊花糖苷、仙茅苷、丹皮酚、哈巴俄苷、五味子醇甲线性范围分别为0.0404~1.616 μg(r =0.9999),0.0194~0.774 μg(r =0.9998),0.0202 ~0.810μg(r =0.9997),0.0201~0.804μg(r=0.9999),0.0206~0.826μg(r=0.9998),0.0398~1.592 μg(r=0.9999).平均加样回收率(n=6)分别为97.8%,98.2%,97.9%,98.3%,101.2%,97.5%;RSD分别为1.2%,1.1%,0.95%,1.1%,0.97%,1.1%.结论:该方法操作简单,重复性好,为评价和监控更年安片的质量提供可靠的方法.%Objective;To establish a method for simultaneous determination of six active ingredients(2,3,5,4' -tetrahydroxystilbene -2 - 0 -β - D - glucoside, acteoside, curculigoside, paeonol, harpagoside, schisandrin). Methods; The determination was carried out with Alltima ODS column ( 250 mm x 4. 6 mm, 5 μm); The mobile phase consisted of acetonitrile( A) -0. 05% phosphoric acid solution( B) with gradient elution at a flow rate of 1. 0 Ml · min-1; The detection wavelengths were 254 nm for schisandrin, 284 nm for curculigoside, paeonol and harpagoside, 330 nm for 2,3,5,4' -tetrahydroxystilbene -2 - 0 -β - D - glucoside and acteoside. ResultS;The calibration curves were linear in the ranges of 0. 0404 -1.616 μg for 2,3,5 ,4' - tetrahydroxystilbene -2 - 0 -0 -D -glucoside(r =0. 9999), 0. 0194 -0. 774 μg for acteoside (r =0. 9998), 0. 0202 -0. 810 |xg for

  13. Simultaneous Determination of Three Active Ingredients in Compound Allantoin VitaminB6-E and Aminoethylsulfonic Acid Eye Drops by HPLC-DAD%HPLC-DAD同时测定复方尿维氨滴眼液中3种活性成分的含量

    Institute of Scientific and Technical Information of China (English)

    金鹏飞; 吴学军; 邹定; 马捷; 姜文清; 郭郁

    2009-01-01

    OBJECTIVE To establish a high performance liquid chromatography-diode array detection (HPLC-DAD) method for the simultaneous determination of chondroitin sulfate sodium, allantoin and vitamin B_6 in Compound Allantoin VitaminB_6-E and Aminoethylsulfonic Acid Eye Drops. METHODS An Alltima C_(18) column (4.6 mm×250 mm, 5 μm particle size) was used for the separation at room temperature, with 25 mmol-L~(-1) ammonium dihydrogen phosphate (containing 0.01% heptanesulfonic acid sodium salt)-acetonitrile (95:5) as the mobile phase at the flow rate of 0.5 mL·min~(-1). The detection wavelengthes for chondroitin sulfate sodium, allantoin and vitamin B_6 were 195, 215 and 291 ran, respectively. RESULTS The method showed good linearity for chondroitin sulfate sodium, allantoin and vitamin B_6, with all correlation coefficients (r) greater than 0.999 6. The specificity study showed satisfactory resolutions between chondroitin sulfate sodium, allantoin, vitamin B_6 other ingredients and forced degradation products. The precisions and stability were satisfactory with all relative standard deviations (RSD) of peak areas lower than 2.0%, and the spiked recovery of three ingredients ranged from 99.01% to 101.92%. CONCLUSION This method is an accurate, fast and simple method for the simultaneous determination of chondroitin sulfate sodium, allantoin and vitamin B6 in Compound Allantoin VitaminB_6-E and Aminoethylsulfonic Acid Eye Drops.%目的 应用高效液相色谱-二极管阵列检测器(HPLC-DAD),建立复方尿维氨滴眼液中硫酸软骨素钠,尿囊素和维生素B6含量的同时测定方法.方法 采用Alltima C_(18)色谱柱(4.6 mm×250 mm,5μm),以25 mmol·L~(-1)磷酸二氢铵(含0.01%辛烷磺酸钠)-乙睛(95:5)为流动相,流速0.5 mL·min~(-1),室温测定,硫酸软骨素钠,尿囊素和维生素B6的检测波长分别为195,215和291mm.结果 硫酸软骨素钠,尿囊素和维生素B6的线性关系良好,线性相关系数(r)都大于0.999 6;专属性强,3

  14. Evaluation of Suwannee River NOM electrophoretic fractions by RP-HPLC with online absorbance and fluorescence detection.

    Science.gov (United States)

    Trubetskaya, Olga E; Richard, Claire; Trubetskoj, Oleg A

    2015-07-01

    Reversed-phase high-performance liquid chromatography (RP-HPLC) with online absorbance and fluorescence detections was used for the evaluation of Suwannee River natural organic matter (SRNOM) and its fractions A, B, and C+D, obtained by conventional size exclusion chromatography-polyacrylamide gel electrophoresis (SEC-PAGE) setup, for which the electrophoretic mobility (EM) and the absorptivity varied in the order C+D > B > A, and the molecular size (MS) in the opposite order. Analysis of SRNOM and its fractions in part of their relative irreversible adsorption on C18-column and relative distribution of eluted from the column matter on hydrophobic and hydrophilic peaks showed that hydrophobicity of fractions decreased in order: A > B > C+D. The online fluorescence detection showed that SRNOM and its fractions contained at least three groups of humic substances (HS)-like fluorophores with emission maxima at 435, 455-465, and 455/420 nm and two protein-like fluorophores with emission maxima at around 300 and 340 nm. The HS-like fluorophore with emission maximum at 435 nm was located in the hydrophilic peak in all the samples. Those with maxima at 455-465 nm were detected in hydrophobic peaks of fractions A and B. SEC-PAGE setup followed by RP-HPLC allowed us to develop new approach of SRNOM separation on less heterogeneous compounds mixture for their further study and structural identification.

  15. Arsenic speciation analysis by HPLC postcolumn hydride generation and detection by atomic fluorescence spectrometry

    OpenAIRE

    Marschner, K; Musil, S. (Stanislav); Rychlovský, P.; Dědina, J. (Jiří)

    2014-01-01

    The aim of this contribution is to present a new method of hydride generation that enables to generate arsines from iAs , iAs , MMA and DMA in a flow injection mode with the same efficiency and in the next step connection of this hydride generator with HPLC column.

  16. Determination of amygdalin, hydroxysafflor yellow A, paeoniflorin, ferulic acid, salvianolic acid B, and tanshinoneⅡA in Naoxueshuan Tablets by HPLC-DAD%HPLC-DAD法测定脑血栓片中苦杏仁苷、羟基红花黄色素A、芍药苷、阿魏酸、丹酚酸B和丹参酮ⅡA

    Institute of Scientific and Technical Information of China (English)

    周军; 张蕾; 王杰

    2015-01-01

    Objective To develop an HPLC-DAD method for determination of amygdalin, hydroxysafflor yellow A, paeoniflorin, ferulic acid, salvianolic acid B, and tanshinoneⅡA in Naoxueshuan Tablets. Methods HPLC-DAD chromatography was used. The analysis was carried out on an Agilent Poroshell 120 SB-C18 column (100 mm × 4.6 mm, 2.7μm) with methanol - 0.2%phosphoric acid as mobile phases at the flow rate of 0.7 mL/min for gradient elution. Detection with variable wavelength were used, and set at 210 nm for amygdalin, 403 nm for hydroxysafflor yellow A, 230 nm for paeoniflorin, 321 nm for ferulic acid, 278 nm for salvianolic acid, and 270 nm for tanshinoneⅡA. The column temperature was 30℃ with injection volume of 3 —5 μL. Results The linear ranges of six components were 11.90—1158.90, 9.14—91.39, 11.70—1 173.50, 4.04—1 011.00, 3.97—992.20, 4.40—551.00 ng, respectively. The average recoveries were 96.47%, 96.92%, 99.96%, 97.20%, 97.57%, and 96.50%with RSD1.3%, 1.6%, 1.3%, 1.7%, 1.9%, and 0.7%, respectively. Conclusion This method can be used to simultaneously determine amygdalin, hydroxysafflor yellow A, paeoniflorin, ferulic acid, salvianolic acid B, and tanshinoneⅡA in Naoxueshuan Tablets.%目的:建立测定脑血栓片中苦杏仁苷、羟基红花黄色素A、芍药苷、阿魏酸、丹酚酸B和丹参酮ⅡA的HPLC-DAD法。方法采用HPLC-DAD法,Agilent Poroshell 120 SB-C18色谱柱(100 mm×4.6 mm,2.7μm);流动相:甲醇–0.2%磷酸溶液,梯度洗脱;检测波长分别为210 nm(苦杏仁苷)、403 nm(羟基红花黄色素A)、230 nm(芍药苷)、321 nm(阿魏酸)、286 nm(丹酚酸B)、270 nm(丹参酮ⅡA);体积流量:0.7 mL/min;柱温:30℃;进样量3~5μL。结果苦杏仁苷、羟基红花黄色素A、芍药苷、阿魏酸、丹酚酸B和丹参酮ⅡA 6个成分的线性范围分别为11.90~1158.90、9.14~91.39、11.70~1173.50、4.04~1011.00、3.97~992.20、4.40~551.00 ng

  17. Lab with Dad

    Science.gov (United States)

    Havers, Brenda; Delmotte, Karen

    2012-01-01

    Family science nights are fantastic, but planning one can be overwhelming, especially when one considers the already overloaded schedule of a classroom teacher. To overcome this challenge, the authors--colleagues with a mutual love of science--developed a much simpler annual event called "Lab With Dad." The purpose was for one target age group of…

  18. Oxidative stress during bacterial growth characterized through microdialysis sampling coupled with HPLC/fluorescence detection of malondialdehyde.

    Science.gov (United States)

    Hsu, Keng-Chang; Hsu, Pi-Fu; Chen, Ya-Ching; Lin, Hsin-Chieh; Hung, Chih-Chang; Chen, Po-Chih; Huang, Yeou-Lih

    2016-04-15

    Organisms that grow aerobically are routinely exposed to oxidative stress in the form of reactive oxygen species. Monitoring the dynamic variations of oxidative stress allows us to understand its role in basic cellular function and determine mechanisms of antioxidation. In this study, microdialysis (MD) sampling was employed for continuous monitoring of the formation of malondialdehyde (MDA) in a bacterium-inoculated culture broth. To test the practicality of this approach, oxidative stress was induced by cadmium and then a 60-min interval was selected to collect sufficient amounts of dialysate for high-performance liquid chromatography with fluorescence (HPLC-FL) detection. After optimization of this simple-to-operate, simultaneous, and continuous method for dynamic monitoring of MDA during periods of bacterial growth, a retrodialysis technique and a no-net-flux method were used to assess the probe recovery and analytical performance of the proposed system. The mean probe recovery of MDA was 78.6 ± 0.9%, with intra- and interday precisions of 2.7-6.1 and 3.5-7.6%, respectively. To evaluate the practicality of this method, the dynamic variations in the concentrations of MDA in standardized bacterial species (Staphylococcus aureus, ATCC(®) 29213™) were monitored continuously for 24h. The analytical results confirmed that this MD sampling technique combined with HPLC-FL detection can be used to accurately and continuously monitor the levels of MDA in microbially inoculated culture broths.

  19. Application of HPLC combined with laser induced fluorescence for protein profile analysis of tissue homogenates in cervical cancer.

    Science.gov (United States)

    Bhat, Sujatha; Patil, Ajeetkumar; Rai, Lavanya; Kartha, V B; Chidangil, Santhosh

    2012-01-01

    A highly objective method, High Performance Liquid Chromatography with Laser Induced Fluorescence (HPLC-LIF) technique was used to study the protein profiles of normal and cervical cancer tissue homogenates. A total of 44 samples including normal cervical biopsy samples from the hysterectomy patients and the patients suffering from different stages of the cervical cancer were recorded by HPLC-LIF and analysed by Principle Component Analysis (PCA) to get statistical information on different tissue components. Discrimination of different stages of the samples was carried out by considering three parameters--scores of factor, spectral residual, and Mahalanobis Distance. Diagnostic accuracy of the method was evaluated using Receiver Operating Characteristic (ROC) analysis, and Youden's index (J) plots. The PCA results showed high sensitivity and specificity (~100) for cervical cancer diagnosis. ROC and Youden's index curves for both normal and malignant standard sets show good diagnostic accuracy with high AUC values. The statistical analysis has shown that the differences in protein profiles can be used to diagnose biochemical changes in the tissue, and thus can be readily applied for the detection of cervical cancer, even in situations where a histopathology examination is not easy because of nonavailability of experienced pathologists.

  20. Application of HPLC Combined with Laser Induced Fluorescence for Protein Profile Analysis of Tissue Homogenates in Cervical Cancer

    Directory of Open Access Journals (Sweden)

    Sujatha Bhat

    2012-01-01

    Full Text Available A highly objective method, High Performance Liquid Chromatography with Laser Induced Fluorescence (HPLC-LIF technique was used to study the protein profiles of normal and cervical cancer tissue homogenates. A total of 44 samples including normal cervical biopsy samples from the hysterectomy patients and the patients suffering from different stages of the cervical cancer were recorded by HPLC-LIF and analysed by Principle Component Analysis (PCA to get statistical information on different tissue components. Discrimination of different stages of the samples was carried out by considering three parameters—scores of factor, spectral residual, and Mahalanobis Distance. Diagnostic accuracy of the method was evaluated using Receiver Operating Characteristic (ROC analysis, and Youden's index (J plots. The PCA results showed high sensitivity and specificity (∼100 for cervical cancer diagnosis. ROC and Youden's index curves for both normal and malignant standard sets show good diagnostic accuracy with high AUC values. The statistical analysis has shown that the differences in protein profiles can be used to diagnose biochemical changes in the tissue, and thus can be readily applied for the detection of cervical cancer, even in situations where a histopathology examination is not easy because of nonavailability of experienced pathologists.

  1. Determination of Trace Level of cAMP in Locusta Migratoria Manilensis Meyen by HPLC with Fluorescence Derivation

    Directory of Open Access Journals (Sweden)

    Canping Pan

    2006-08-01

    Full Text Available A sensitive and rapid method was developed for the determination of cAMP inLocusta migratoria manilensis Meyen by high-performance liquid chromatography withfluorescence detection. The cAMP was derivatized using chloroacetaldehyde and TBASbuffer/methanol was used as the mobile phase. A detection quantification of 40 fmol/mlcould be achieved when using fluorescence detection. An HPLC-MS method using DMHAas an ion-pair agent to analyze cAMP was also demonstrated. We studied the effect ofdopamine and other stimulants on cAMP levels from isolated locust central nervoussystems. The new method is well suited for the analysis of cAMP in small biologicalsamples.

  2. Metallothionein induction by Cu, Cd and Hg in Dicentrarchus labrax liver: assessment by RP-HPLC with fluorescent detection and spectrophotometry

    OpenAIRE

    Jebali, Jamel; Banni, Mohamed; Gerbej, Hamadi; Boussetta, Hamadi; López-Barea, Juan; Alhama, José

    2008-01-01

    Metallothionein induction by Cu, Cd and Hg in Dicentrarchus labrax liver: assessment by RP-HPLC with fluorescent detection and spectrophotometry correspondance: Corresponding author. Tel.: +34 957 218082; fax: +34 957 218688. (Alhama, Jose) (Alhama, Jose) Laboratoire de Biochimie et de Toxicologie Environmentale--> , Institut Superieur d?Agronomie de Chott-Mariem--> , 4042 Sousse--> - TUNISIA (Jebali,...

  3. Development of a new HPLC method using fluorescence detection without derivatization for determining purine nucleoside phosphorylase activity in human plasma.

    Science.gov (United States)

    Giuliani, Patricia; Zuccarini, Mariachiara; Buccella, Silvana; Rossini, Margherita; D'Alimonte, Iolanda; Ciccarelli, Renata; Marzo, Matteo; Marzo, Antonio; Di Iorio, Patrizia; Caciagli, Francesco

    2016-01-15

    Purine nucleoside phosphorylase (PNP) activity is involved in cell survival and function, since PNP is a key enzyme in the purine metabolic pathway where it catalyzes the phosphorolysis of the nucleosides to the corresponding nucleobases. Its dysfunction has been found in relevant pathological conditions (such as inflammation and cancer), so the detection of PNP activity in plasma could represent an attractive marker for early diagnosis or assessment of disease progression. Thus the aim of this study was to develop a simple, fast and sensitive HPLC method for the determination of PNP activity in plasma. The separation was achieved on a Phenomenex Kinetex PFP column using 0.1% formic acid in water and methanol as mobile phases in gradient elution mode at a flow rate of 1ml/min and purine compounds were detected using UV absorption and fluorescence. The analysis was fast since the run was achieved within 13min. This method improved the separation of the different purines, allowing the UV-based quantification of the natural PNP substrates (inosine and guanosine) or products (hypoxanthine and guanine) and its subsequent metabolic products (xanthine and uric acid) with a good precision and accuracy. The most interesting innovation is the simultaneous use of a fluorescence detector (excitation/emission wavelength of 260/375nm) that allowed the quantification of guanosine and guanine without derivatization. Compared with UV, the fluorescence detection improved the sensitivity for guanine detection by about 10-fold and abolished almost completely the baseline noise due to the presence of plasma in the enzymatic reaction mixture. Thus, the validated method allowed an excellent evaluation of PNP activity in plasma which could be useful as an indicator of several pathological conditions.

  4. Determination of Angiotensin-(1-7) with HPLC/Fluorescence-Detection.

    Science.gov (United States)

    Russ, Miriam; Hauser, Susanne; Wintersteiger, Reinhold; Greilberger, Joachim; Andrä, Michaela; Ortner, Astrid

    2016-01-01

    Angiotensin-(1-7) is an important active component in the renin-angiotensin-system. Due to its cardio protective effects it is now under investigation in combination with antioxidants as a reperfusion solution. The combination showed impressive effects on isolated hearts of male Wistar rats after induced ischemia. In this work a high performance liquid chromatography method with fluorescence detection was developed for the first time for in-process measurements as well as for stability tests of the peptide in the novel antioxidant-containing Karal® solution. For fluorescence detection of angiotensin-(1-7) fluorescamine as derivatization dye was applied. Under optimized conditions the method showed linearity over the range of 50 to 5000 ng/mL with R(2) of 0.9988 and an overall precision better than 5.0 %. LOD and LOQ were determined to be in the femtomol range on column. It was found that stability of angiotensin-(1-7) could be significantly improved in the antioxidant containing preparation compared to aqueous solutions.

  5. Simultaneous Determination of Thirteen Chemical Materials Illegally Added in Antirheumatic Health Products by HPLC-DAD%HPLC-DAD法同时测定抗风湿类保健食品中非法添加的13种化学成分

    Institute of Scientific and Technical Information of China (English)

    金舒

    2016-01-01

    Objective: To simultaneously detect 13 anti-rheumatic chemical components added illegally in health food. Methods: Using HPLC-DAD, the separation was performed on the XBridge C18 (4.6 mm×250 mm, 5μm) column by gradient elution at a flow rate of 1.0 mL·min-1. The mobile phase consisted of ace-tonitril-0.02 mol·L-1 ammonium acetate (including 0.075% acetic acid) and the detection wavelength was 230 nm. Results: The correlation coefficients of the equation of linear regression were above 0.9999. The average recoveries were 95.6%-102.5% and the relative standard deviations were less than 1.6% (n=3). U-PLC-QTOF-MS was used to further confirm. Conclusion: The method is simple and reliable, and can be used for simultaneously detecting 13 anti-rheumatic chemical components (acetaminophen, aspirin, trimetho-prim, aminopyrine, hydrocortisone, piroxicam, dexamethasone, naproxen, prednisone acetate, diclofenac sodi-um, indometacin, phenylbutazone and ibuprofen) in health food.%目的:同时检测抗风湿类保健食品中非法添加的13种化学成分。方法:采用高效液相色谱串联二极管阵列检测器法,使用XBridge C18(4.6 mm×250 mm,5μm)色谱柱进行分离,流动相为乙腈-0.02 mol·L-1醋酸铵(含0.075%乙酸),梯度洗脱,流速为1.0 mL·min-1,检测波长为230 nm。结果:各化学成分线性回归方程的相关系数均大于0.9999,平均回收率为95.6%~102.5%, RSD均小于1.6%(n=3),并用UPLC-QTOF-MS法进一步确证。结论:该方法操作简便可靠,能同时测定抗风湿类保健食品中非法添加的13种化学成分,分别为对乙酰氨基酚、阿司匹林、甲氧苄啶、氨基比林、氢化可的松、吡罗昔康、地塞米松、萘普生、醋酸泼尼松、双氯芬酸钠、吲哚美辛、保泰松、布洛芬。

  6. Fluorescent adduct formation with terbium: a novel strategy for transferrin glycoform identification in human body fluids and carbohydrate-deficient transferrin HPLC method validation.

    Science.gov (United States)

    Sorio, Daniela; De Palo, Elio Franco; Bertaso, Anna; Bortolotti, Federica; Tagliaro, Franco

    2017-02-01

    This paper puts forward a new method for the transferrin (Tf) glycoform analysis in body fluids that involves the formation of a transferrin-terbium fluorescent adduct (TfFluo). The key idea is to validate the analytical procedure for carbohydrate-deficient transferrin (CDT), a traditional biochemical serum marker to identify chronic alcohol abuse. Terbium added to a human body-fluid sample produced TfFluo. Anion exchange HPLC technique, with fluorescence detection (λ exc 298 nm and λ em 550 nm), permitted clear separation and identification of Tf glycoform peaks without any interfering signals, allowing selective Tf sialoforms analysis in human serum and body fluids (cadaveric blood, cerebrospinal fluid, and dried blood spots) hampered for routine test. Serum samples (n = 78) were analyzed by both traditional absorbance (Abs) and fluorescence (Fl) HPLC methods and CDT% levels demonstrated a significant correlation (p body fluid analysis. Its sensitivity and absence of interferences extend clinical applications being reliable for CDT assay on body fluids usually not suitable for routine test. Graphical Abstract The formation of a transferrin-terbium fluorescent adduct can be used to analyze the transferrin glycoforms. The HPLC method for carbohydrate-deficient transferrin (CDT%) measurement was validated and employed to determine the levels in different body fluids.

  7. HPLC法测定山茱萸生品中5-羟甲基糠醛、莫诺苷和马钱苷含量%Simultaneous determination of 5-HMF,morroniside and loganin in Cornus officinalis by HPLC-DAD

    Institute of Scientific and Technical Information of China (English)

    刘少静; 杨黎彬; 赵宁; 王小库; 朱改改

    2011-01-01

    目的 采用高效液相二极管阵列检测法(HPLC-DAD)同时测定山茱萸生品中3个成分的含量:5-羟甲基糠醛(5-HMF)、莫诺苷和马钱苷.方法 采用Venusil ABS C18色谱柱(250 mm × 4.6 mm,5 μm),以甲醇和水为流动相,梯度洗脱,流速 1 ml·min-1,柱温 30 ℃,检测波长284、240、238 nm.结果 5-HMF 、莫诺苷和马钱苷的峰面积与浓度的线性关系均良好(r≥0.999 6);加样回收率分别为100%、100.62%、100.53%.结论 该方法快速、准确、重现性好,可用于同时测定山茱萸生品中5-羟甲基糠醛、莫诺苷和马钱苷的含量.%Aim To estahlish a high performance liquid chromatographic method with diode array detection( HPLC-DAD )for simultaneous determination of 5-HMF,morroniside and loganin in Cornus officinalis. Methods Three constituents were analyzed simultaneously with a Venusil ABS C18 column( 250 mm ×4. 6 mm,5 μm )by gradient elution using methanol-water as the mobile phase. The column was maintained at 30℃ . The flow rate was 1 ml · min -1. The detection wavelength was set at 284 ,240 and 238 nm. Results All the constituents showed good linearity( r≥ 0. 9996 )in the range of the tested concentration. The average recoveries were 100%, 100. 62%,100. 53% . Conclusion The validated method has the advantages of quickness, precision and reliahility , allowing its simultaneous determination of 5-HMF, morroniside and loganin in Cornus officinalis.

  8. [Development of Determination Method of Fluoroquinolone Antibiotics in Sludge Based on Solid Phase Extraction and HPLC-Fluorescence Detection Analysis].

    Science.gov (United States)

    Dai, Xiao-hu; Xue, Yong-gang; Liu, Hua-jie; Dai, Ling-ling; Yan, Han; Li, Ning

    2016-04-15

    Fluoroquinolone antibiotics (FQs), as the common pharmaceuticals and personal care products (PPCPs), are widespread in the environment. FQs contained in wastewater would be ultimately enriched in sludge, posing a potential threat to the consequent sludge utilization. To optimize the analytical method applicable to the determination of FQs in sludge, the authors selected ofloxacin (OFL), norfioxacin (NOR), ciprofloxacin (CIP) and lomefloxacin (LOM) as the target FQs, and established a method which was based on cell lysis, FQs extraction with triethylamine/methanol/water solution, Solid Phase Extraction (SPE) and HPLC-Fluorescence Detection (FLD) determination. After the investigation, phosphoric acid-triethylamine was decided to be the buffer salt, and methanol was chosen as the organic mobile phase. The gradient fluorescence scanning strategy was proved to be necessary for the optimal detection as well. Furthermore, by the designed orthogonal experiments, the effects of the extraction materials, pH, and the eluents on the efficiency of SPE extraction were evaluated, by which the optimal extraction conditions were determined. As a result, FQs in liquid samples could be analyzed by utilizing HLB extraction cartridge, and the recovery rates of the four FQs were in the range of 82%-103%. As for solid samples, the recovery rates of the four FQs contained reached up to 71%-101%. Finally, the adsorptivity of the sludge from the different tanks ( anaerobic, anoxic and oxic tanks) was investigated, showing gradual decrease in the adsorption capacity, but all adsorbed over 90% of the EQs. This conclusion also confirmed that 50% removal of FQs in the domestic wastewater treatment plant was realized by sludge adsorption.

  9. Analysis of fish bile with HPLCfluorescence to determine environmental exposure to benzo(a)pyrene

    Science.gov (United States)

    Johnston, Eric P.; Baumann, Paul C.

    1989-01-01

    Brown bullhead from the Black River, Ohio, have a high incidence of liver neoplasia which is associated with elevated concentrations of polynuclear aromatic hydrocarbons (PAHs) in the sediment. We evaluated the use of biliary concentrations of benzo(a)pyrene [B(a)P] equivalents as a means for determining PAH exposure. Bile was collected from 16 brown bullheads and 8 common carp taken from each of two Lake Erie tributaries in Ohio, the industrialized Black River and the non-industrialized Old Woman Creek. Hatchery bullhead (n = 8) were used to determine base levels of PAHs. A high performance liquid chromatography (HPLC) — fluorescence technique was used to determine the concentration of B(a)P equivalents in the bile samples. The area of all peaks fluorescing at 380/430 nm was summed to give a single value for B(a)P equivalents in each sample. Concentrations of B(a)P equivalents generally reflected concentrations of PAH in sediment where fish were collected. Bile taken from Black River carp contained the highest concentration of B(a)P equivalents and was significantly different from all other groups. The value obtained for Black River bullhead was also high and was found to be significantly different from hatchery bullhead. B(a)P equivalents varied between carp and bullhead from the same habitat possibly because of differing food habits or metabolic pathways. However, our results indicate that relative levels of B(a)P equivalents in the bile of fish correspond well to B(a)P levels in sediment and may offer a means of determining environmental exposure of fish to the parent compound.

  10. Development of an analytical method for antimony speciation in vegetables by HPLC-hydride generation-atomic fluorescence spectrometry.

    Science.gov (United States)

    Olivares, David; Bravo, Manuel; Feldmann, Jorg; Raab, Andrea; Neaman, Alexander; Quiroz, Waldo

    2012-01-01

    A new method for antimony speciation in terrestrial edible vegetables (spinach, onions, and carrots) was developed using HPLC with hydride generation-atomic fluorescence spectrometry. Mechanical agitation and ultrasound were tested as extraction techniques. Different extraction reagents were evaluated and optimal conditions were determined using experimental design methodology, where EDTA (10 mmol/L, pH 2.5) was selected because this chelate solution produced the highest extraction yield and exhibited the best compatibility with the mobile phase. The results demonstrated that EDTA prevents oxidation of Sb(III) to Sb(V) and maintains the stability of antimony species during the entire analytical process. The LOD and precision (RSD values obtained) for Sb(V), Sb(III), and trimethyl Sb(V) were 0.08, 0.07, and 0.9 microg/L and 5.0, 5.2, and 4.7%, respectively, for a 100 microL sample volume. The application of this method to real samples allowed extraction of 50% of total antimony content from spinach, while antimony extracted from carrots and onion samples ranged between 50 and 60 and 54 and 70%, respectively. Only Sb(V) was detected in three roots (onion and spinach) that represented 60-70% of the total antimony in the extracts.

  11. Structural requirements for Caenorhabditis elegans DcpS substrates based on fluorescence and HPLC enzyme kinetic studies.

    Science.gov (United States)

    Wypijewska, Anna; Bojarska, Elzbieta; Stepinski, Janusz; Jankowska-Anyszka, Marzena; Jemielity, Jacek; Davis, Richard E; Darzynkiewicz, Edward

    2010-07-01

    The activity of the Caenorhabditis elegans scavenger decapping enzyme (DcpS) on its natural substrates and dinucleotide cap analogs, modified with regard to the nucleoside base or ribose moiety, has been examined. All tested dinucleotides were specifically cleaved between beta- and gamma-phosphate groups in the triphosphate chain. The kinetic parameters of enzymatic hydrolysis (K(m), V(max)) were determined using fluorescence and HPLC methods, as complementary approaches for the kinetic studies of C. elegans DcpS. From the kinetic data, we determined which parts of the cap structure are crucial for DcpS binding and hydrolysis. We showed that m(3)(2,2,7)GpppG and m(3)(2,2,7)GpppA are cleaved with higher rates than their monomethylated counterparts. However, the higher specificity of C. elegans DcpS for monomethylguanosine caps is illustrated by the lower K(m) values. Modifications of the first transcribed nucleotide did not affect the activity, regardless of the type of purine base. Our findings suggest C. elegans DcpS flexibility in the first transcribed nucleoside-binding pocket. Moreover, although C. elegans DcpS accommodates bulkier groups in the N7 position (ethyl or benzyl) of the cap, both 2'-O- and 3'-O-methylations of 7-methylguanosine result in a reduction in hydrolysis by two orders of magnitude.

  12. Simultaneous determination of doxorubicin and its dipeptide prodrug in mice plasma by HPLC with fluorescence detection

    Directory of Open Access Journals (Sweden)

    Jing Han

    2016-06-01

    Full Text Available A simple and sensitive high performance liquid chromatography with fluorescence detection (HPLC–FD has been developed for simultaneous quantification of doxorubicin (DOX and its dipeptide conjugate prodrug (PDOX in mice plasma. The chromatographic separation was carried out on an Amethyst C18–H column with gradient mobile phase of 0.1% formic acid and 0.1% formic acid in acetonitrile at a flow rate of 1.0 mL/min. The excitation and emission wavelengths were set at 490 and 550 nm, respectively. The method was comprehensively validated. The limits of detection were low up to 5.0 ng/mL for DOX and 25.0 ng/mL for PDOX. And the limits of quantification were low up to 12.5 ng/mL for DOX and 50 ng/mL for PDOX, which were lower than those for most of the current methods. The calibration curves showed good linearity (R2>0.999 over the concentration ranges. The extraction recoveries ranged from 84.0% to 88.2% for DOX and from 85.4% to 89.2% for PDOX. Satisfactory intra-day and inter-day precisions were achieved with RSDs less than 9.1%. The results show that the developed HPLC–FD method is accurate, reliable and will be helpful for preclinical pharmacokinetic study of DOX and PDOX.

  13. Simultaneous determination of doxorubicin and its dipeptide prodrug in mice plasma by HPLC with fluorescence detection$

    Institute of Scientific and Technical Information of China (English)

    Jing Han; Jue Zhang; Haiyan Zhao; Yan Li; Zilin Chen

    2016-01-01

    A simple and sensitive high performance liquid chromatography with fluorescence detection (HPLC–FD) has been developed for simultaneous quantification of doxorubicin (DOX) and its dipeptide conjugate prodrug (PDOX) in mice plasma. The chromatographic separation was carried out on an Amethyst C18–H column with gradient mobile phase of 0.1%formic acid and 0.1%formic acid in acetonitrile at a flow rate of 1.0 mL/min. The excitation and emission wavelengths were set at 490 and 550 nm, respectively. The method was comprehensively validated. The limits of detection were low up to 5.0 ng/mL for DOX and 25.0 ng/mL for PDOX. And the limits of quantification were low up to 12.5 ng/mL for DOX and 50 ng/mL for PDOX, which were lower than those for most of the current methods. The calibration curves showed good linearity (R2 4 0.999) over the concentration ranges. The extraction recoveries ranged from 84.0%to 88.2% for DOX and from 85.4% to 89.2% for PDOX. Satisfactory intra-day and inter-day precisions were achieved with RSDs less than 9.1%. The results show that the developed HPLC–FD method is accurate, reliable and will be helpful for preclinical pharmacokinetic study of DOX and PDOX.

  14. Rapid Determination of Costunolide and Dehydrocostuslactone in Human Plasma Sample and Chinese Patent Medicine Xiang Sha Yang Wei Capsule Using HPLC-DAD Coupled with Second-order Calibration

    Institute of Scientific and Technical Information of China (English)

    刘亚娟; 吴海龙; 朱绍华; 康超; 许慧; 苏志义; 谷惠文; 俞汝勤

    2012-01-01

    A novel methodology that combines high performance liquid chromatography with photodiode-array detector (HPLC-DAD) coupled with second-order calibration method based on alternating trilinear decomposition (ATLD) algorithm was used in determination of the effective constituents such as costunolide and dehydrocostuslactone, in plasma sample and Chinese patent medicine Xiang Sha Yang Wei (XSYW) capsule. Complicated systems such as plasma and Chinese patent medicine which have intricate components are tedious to isolate and purify. The problem that chromatographic peaks are heavily overlapped among the analytes and interferents from the background matrices can be resolved, and the satisfactory quantification results have been gained with the help of the ATLD algorithm which utilized "mathematical separation" instead of partial "physical or chemical separation". Meanwhile, HPLC-MS/MS method was used to validate the accuracy of the proposed determination method.

  15. Immunoassay screening of lysergic acid diethylamide (LSD) and its confirmation by HPLC and fluorescence detection following LSD ImmunElute extraction.

    Science.gov (United States)

    Grobosch, T; Lemm-Ahlers, U

    2002-04-01

    In all, 3872 urine specimens were screened for lysergic acid diethylamide (LSD) using the CEDIA DAU LSD assay. Forty-eight samples, mainly from psychiatric patients or drug abusers, were found to be LSD positive, but only 13 (27%) of these could be confirmed by high-performance liquid chromatography with fluorescence detection (HPLC-FLD) following immunoaffinity extraction (IAE). Additional analysis for LSD using the DPC Coat-a-Count RIA was performed to compare the two immunoassay screening methods. Complete agreement between the DPC RIA assay and HPLC-FLD results was observed at concentrations below a cutoff concentration of 500 pg/mL. Samples that were LSD positive in the CEDIA DAU assay but not confirmed by HPLC-FLD were also investigated for interfering compounds using REMEDI HS drug-profiling system. REMEDI HS analysis identified 15 compounds (parent drugs and metabolites) that are believed to cross-react in the CEDIA DAU LSD assay: ambroxol, prilocaine, pipamperone, diphenhydramine, metoclopramide, amitriptyline, doxepine, atracurium, bupivacaine, doxylamine, lidocaine, mepivacaine, promethazine, ranitidine, and tramadole. The IAE/HPLC-FLD combination is rapid, easy to perform and reliable. It can reduce costs when standard, rather than more advanced, HPLC equipment is used, especially for labs that perform analyses for LSD infrequently. The chromatographic analysis of LSD, nor-LSD, and iso-LSD is not influenced by any of the tested cross-reacting compounds even at a concentration of 100 ng/mL.

  16. Determination of Bergamot lactone and Isopimpinellin in Cnidium monnieri by HPLC-DAD%蛇床子中佛手柑内酯和异虎耳草素含量的高效液相色谱-二级管阵列检测法同时测定

    Institute of Scientific and Technical Information of China (English)

    仝立国; 谢君; 冯玛莉

    2014-01-01

    目的 建立同时测定蛇床子中佛手柑内酯和异虎耳草素含量的方法.方法 采用高效液相色谱(HPLC)法对其含量进行测定,色谱柱为Sino Chrom C18 (250mm×4.6mm,5μm,Elite),流动相为甲醇-0.1%醋酸(55∶45),流速1.0ml/min,柱温25℃;二极管阵列检测器(DAD)检测器,检测波长310hm.结果 佛手柑内酯在0.031 5~0.945 μg范围内呈良好的线性关系A=2385C-6.666 (r=0.9999),异虎耳草素在0.029 6~0.888μg范围内呈良好的线性关系A=1711C+1.203(r=0.999 9);平均回收率分别为99.2%和99.1%,相对标准偏差分别为0.91%和0.67%(n=6);在常温下8h内稳定性较好.结论 HPLC-DAD法简便快速,准确,重现性好,结果可靠,可作为蛇床子的质量控制方法.

  17. Determination of propofol concentration in blood by HPLC with fluorescence detection%HPLC-荧光检测测定人全血中丙泊酚浓度

    Institute of Scientific and Technical Information of China (English)

    单国瑾; 吴新民; 苏玉; 涂恩平

    2001-01-01

    目的建立反相高效液相色谱(HPLC)荧光检测测定人全血中丙泊酚浓度的方法.方法用环己烷提取全血中的丙泊酚,以百里酚为内标定量,用HPLC-荧光检测器测定丙泊酚含量.结果本法测定的线性范围为5~3000ng/ml,该方法回收率高,重现性好.结论本方法灵敏,选择性强,快速、准确地测出全血中微量丙泊酚的含量,适于研究需要.

  18. Determination of Citalopram in Human Plasma by HPLC with Fluorescence Detection%人血浆中西酞普兰的HPLC-荧光法测定

    Institute of Scientific and Technical Information of China (English)

    张永泰; 张淑慧; 汪玉梅; 郭文敏

    2004-01-01

    建立了HPLC-荧光法测定人血浆中西酞普兰的浓度.血浆样品经液-液萃取后测定,用Zorbax SB C8色谱柱,磷酸二氢铵缓冲液(pH 3.5)-乙腈(65:35)为流动相,激发波长240 nm,发射波长302 nm.血浆中西酞普兰线性范围为1~100ng/ml,最低定量浓度为1ng/ml,日内、日间RSD均小于4.0%,方法平均回收率为93.2%.

  19. Quantitative determination of flavonoids of pickles by HPLC with fluorescence detector%腌菜中黄酮的HPLC-荧光分析

    Institute of Scientific and Technical Information of China (English)

    吴波; 张寒俊; 严建芳

    2010-01-01

    建立了高效液相色谱法(HPLC)荧光法测定腌菜中的桑色素、槲皮素,山奈酚和异鼠李素的含量的方法.桑色素、槲皮素,山奈酚和异鼠李素的线性范围及相关系数分别为0.25mg/L~8mg/L,r=0.9993;0.25~8.mg/L,r=0.9998;0.25mg/L~8mg/L,r=9993;0.25mg/L~8mg/L、r=0.9999.平均加样回收率为89.5%~102.3%.

  20. A novel salting-out assisted extraction coupled with HPLC- fluorescence detection for trace determination of vitamin K homologues in human plasma.

    Science.gov (United States)

    Ahmed, Sameh; Mahmoud, Ashraf M

    2015-11-01

    Recently, new physiological roles of vitamin K homologues have been established in the treatment of rheumatoid arthritis, osteoporosis, hepatocellular carcinoma and leukemia. However, relatively high plasma protein binding, low plasma concentrations and occurrences of interfering lipids make accurate determination of vitamin K homologues a challenging task. Therefore, a sensitive and reliable salting-out assisted liquid/liquid extraction (SALLE) method coupled with HPLC-Fluorescence detection was designed for efficient extraction and quantification of trace levels of vitamin K homologues in human plasma. The investigated vitamin K homologues were phylloquinone (PK, vitamin K1), menaquinone-4 (MK-4) and menaquinone-7 (MK-7). The method employed a new efficient fluorescence derivatization reaction using ethanolic solution of stannous chloride in acidic solution to generate highly fluorescent naphthohydroquinone derivatives. Correlation coefficients were more than 0.998 in the concentration ranges of 0.3-100 ng mL(-1) with detection limits of 0.1-0.17 ng mL(-1) in human plasma. The developed HPLC-FL system was successfully applied for sensitive determination of vitamin K homologues in plasma of healthy volunteers. The developed method may provide a valuable tool in the pharmacoinformatic studies concerning the roles of vitamin K homologues.

  1. Identification of schisandrin as a vascular endothelium protective component in YiQiFuMai Powder Injection using HUVECs binding and HPLC-DAD-Q-TOF-MS/MS analysis.

    Science.gov (United States)

    Li, Fang; Tan, Yi-Sha; Chen, Hong-Lin; Yan, Yan; Zhai, Ke-Feng; Li, Da-Peng; Kou, Jun-Ping; Yu, Bo-Yang

    2015-09-01

    YiQiFuMai Powder Injection (YQFM) is a re-developed preparation based on the well-known traditional Chinese medicine formula Sheng-mai-san. It has been widely used for the treatment of cardiovascular disease with definite clinical efficacy in China, but its bioactive molecules remain obscure. In this study, an effective method has been employed as a tool for screening active components in YQFM, using human umbilical vein endothelial cells (HUVECs) extraction and liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (Q-TOF MS/MS). Nine compounds, which could interact with HUVECs, were identified as ginsenosides Rb1, Rc, Rb2, Rd, 20(S)-Rg3, 20(R)-Rg3, Rk1/Rg5 and schisandrin by comparing with reference substances or literature. In vitro assays showed that schisandrin at concentrations of 10-100 μM protected HUVECs from hypoxia/reoxygenation (H/R) injury, increased cell viability, nitric oxide (NO) content and decreased lactate dehydrogenase (LDH) leakage, malonaldehyde (MDA) content and ROS generation. Moreover, schisandrin pretreatment inhibited cell apoptosis, as evidenced by inhibiting activation of caspase-3 and increasing the Bcl-2/Bax ratio. These data indicate that HUVECs biospecific extraction coupled with HPLC-ESI-Q-TOF-MS/MS analysis is a reliable method for screening potential bioactive components from traditional Chinese medicines. Meanwhile, the vascular endothelium protective property of schisandrin might be beneficial for the treatment of cardiovascular disease.

  2. Comparison of SPE/d-SPE and QuEChERS-Based Extraction Procedures in Terms of Fungicide Residue Analysis in Wine Samples by HPLC-DAD and LC-QqQ-MS.

    Science.gov (United States)

    Tuzimski, Tomasz; Rejczak, Tomasz; Pieniążek, Dominika; Buszewicz, Grzegorz; Teresiński, Grzegorz

    2016-11-01

    Two different extraction and clean-up protocols, based on either the SPE/dispersive SPE (d-SPE) or the quick, easy, cheap, effective, rugged, and safe approach, were optimized and compared for determination of six selected fungicides (benalaxyl, metalaxyl, triadimenol, tebuconazole, diniconazole, and epoxiconazole) in wine samples. The pilot study was performed by applying HPLC with diode-array detection, and optimized procedures were easily transferred to the LC triple-quadrupole MS system. Both extraction procedures presented good performance for all the analytes, with recoveries in the range of 70-132% and SDs ≤20%. The d-SPE clean-up step included in both procedures allows obtaining colorless extracts with the majority of coextracted matrix compounds removed. LC with electrospray ionization and tandem MS operating in the multiple reaction monitoring mode provide high sensitivity and selectivity for trace analysis. Both developed procedures were evaluated in terms of commercial wine sample analysis. In three wine samples, metalaxyl and tebuconazole residues were detected at concentrations from 0.14 to 30.7 ng/mL. Both approaches showed satisfactory feasibility for fungicide residue analysis in wine samples.

  3. Postpartum Depression Affects New Dads, Too

    Science.gov (United States)

    ... https://medlineplus.gov/news/fullstory_163092.html Postpartum Depression Affects New Dads, Too Certain men at greater ... HealthDay News) -- Men can also suffer from postpartum depression after their baby is born. "Dads want to ...

  4. 高效液相色谱-二极管阵列检测方法结合交替三线性分解算法快速定量测定中药黄芩中的黄芩素和汉黄芩素%Simultaneous Determination of Baicalein and Wogonin in Chinese Medicinal Plant Scutellaria Baicalensis Using HPLC-DAD Coupled、with Alternating Trilinear Decomposition

    Institute of Scientific and Technical Information of China (English)

    许慧; 吴海龙; 刘亚娟; 苏志义; 张娟; 聂重重; 李勇; 俞汝勤

    2011-01-01

    利用高效液相色谱-二极管阵列检测(HPLC-DAD)方法结合化学计量学基于交替三线性分解算法的二阶校正方法,在未知基体干扰共存下,同时测定了中药黄芩中的黄芩素和汉黄芩素的含量.当体系组分数选取为3时,测得黄芩素和汉黄芩素的平均回收率分别为104.6±4.3%和100.7±7.4%.该方法不仅能同时给出2种物质的相对色谱图和相对光谱图,还能同时测定未知干扰存在下的2种物质的浓度.%This paper proposed a novel method for simultaneous determination of baicalein and wogonin in Chinese medicinal plant Scutellaria baicalensis by using high performance liquid chromatography with photodiode-array detection (HPLC-DAD) coupled with second-order calibration method based on the alternating trilinear decomposition algorithm,especially in the presence of matrix interferences.When the core consistency diagnostic suggested N was set to 3,the average recoveries of baicalein and wogonin were 104.6±4.3% and 100.7±7.4%, respectively.

  5. Determination of olmesartan in human plasma by HPLC with fluorescence detection%HPLC-荧光检测法测定人血浆中的奥美沙坦

    Institute of Scientific and Technical Information of China (English)

    刘建芳; 王金; 于洋; 侯艳宁

    2006-01-01

    目的:建立一种测定人血浆中奥美沙坦浓度的HPLC-FLD方法.方法:血浆样品用适量甲醇涡旋混匀后用OASISHLB小柱进行萃取.采用Diamonsil C18(150 mm ×4.6 mm,5 μm)分析柱,以乙腈-甲醇-水-冰醋酸(25:40:35:0.1)为流动相,流速1 mL·min-1.荧光法检测,激发波长260 nm,发射波长370 nm.内标为替米沙坦.结果:本方法的线性范围为0.05~3μg·mL-1,最低定量限0.05μg·mL-1,相对回收率为98.7%~101.7%,日内RSD<7%,日间RSD<10%.结论:本法样品处理简单、选择性好,适用于奥美沙坦酯制剂的人体药代动力学研究.

  6. Application of 10-ethyl-acridine-3-sulfonyl chloride for HPLC determination of aliphatic amines in environmental water using fluorescence and APCI-MS.

    Science.gov (United States)

    You, Jinmao; Zhao, Huaixin; Sun, Zhiwei; Xia, Lian; Yan, Tao; Suo, Yourui; Li, Yulin

    2009-05-01

    A simple, sensitive method for the determination of aliphatic amines based on a sulfonylation reaction using 10-ethyl-acridine-3-sulfonyl chloride (EASC) as pre-column labeling reagent with fluorescence detection and APCI-MS identification has been developed. The labeled derivatives exhibited high stability and were enough to be efficiently analyzed by HPLC with an excitation maximum at lambda(ex) 270 nm and an emission maximum at lambda(em) 430 nm. Identification of derivatives was carried out by online post-column MS in positive-ion mode. Comparing with the widely used 5-dimethylaminonaphthalene-1-sulfonylchloride (Dansyl-Cl), EASC-amine derivatives not only exhibited high fluorescence but also exhibited excellent MS ionizable potential. Detection limits obtained from 0.10 pmol injection, at a S/N of 3, were 4.0-12.7 fmol. The mean intra- and inter-assay precision for all aliphatic amine levels were 0.9995.

  7. An industry consensus study on an HPLC fluorescence method for the determination of (±)-catechin and (±)-epicatechin in cocoa and chocolate products.

    Science.gov (United States)

    Shumow, Laura; Bodor, Alison

    2011-07-05

    This manuscript describes the results of an HPLC study for the determination of the flavan-3-ol monomers, (±)-catechin and (±)-epicatechin, in cocoa and plain dark and milk chocolate products. The study was performed under the auspices of the National Confectioners Association (NCA) and involved the analysis of a series of samples by laboratories of five member companies using a common method. The method reported in this paper uses reversed phase HPLC with fluorescence detection to analyze (±)-epicatechin and (±)-catechin extracted with an acidic solvent from defatted cocoa and chocolate. In addition to a variety of cocoa and chocolate products, the sample set included a blind duplicate used to assess method reproducibility. All data were subjected to statistical analysis with outliers eliminated from the data set. The percent coefficient of variation (%CV) of the sample set ranged from approximately 7 to 15%. Further experimental details are described in the body of the manuscript and the results indicate the method is suitable for the determination of (±)-catechin and (±)-epicatechin in cocoa and chocolate products and represents the first collaborative study of this HPLC method for these compounds in these matrices.

  8. An industry consensus study on an HPLC fluorescence method for the determination of (±-catechin and (±-epicatechin in cocoa and chocolate products

    Directory of Open Access Journals (Sweden)

    Bodor Alison

    2011-07-01

    Full Text Available Abstract Background This manuscript describes the results of an HPLC study for the determination of the flavan-3-ol monomers, (±-catechin and (±-epicatechin, in cocoa and plain dark and milk chocolate products. The study was performed under the auspices of the National Confectioners Association (NCA and involved the analysis of a series of samples by laboratories of five member companies using a common method. Methodology The method reported in this paper uses reversed phase HPLC with fluorescence detection to analyze (±-epicatechin and (±-catechin extracted with an acidic solvent from defatted cocoa and chocolate. In addition to a variety of cocoa and chocolate products, the sample set included a blind duplicate used to assess method reproducibility. All data were subjected to statistical analysis with outliers eliminated from the data set. Results The percent coefficient of variation (%CV of the sample set ranged from approximately 7 to 15%. Conclusions Further experimental details are described in the body of the manuscript and the results indicate the method is suitable for the determination of (±-catechin and (±-epicatechin in cocoa and chocolate products and represents the first collaborative study of this HPLC method for these compounds in these matrices.

  9. An industry consensus study on an HPLC fluorescence method for the determination of (±)-catechin and (±)-epicatechin in cocoa and chocolate products

    Science.gov (United States)

    2011-01-01

    Background This manuscript describes the results of an HPLC study for the determination of the flavan-3-ol monomers, (±)-catechin and (±)-epicatechin, in cocoa and plain dark and milk chocolate products. The study was performed under the auspices of the National Confectioners Association (NCA) and involved the analysis of a series of samples by laboratories of five member companies using a common method. Methodology The method reported in this paper uses reversed phase HPLC with fluorescence detection to analyze (±)-epicatechin and (±)-catechin extracted with an acidic solvent from defatted cocoa and chocolate. In addition to a variety of cocoa and chocolate products, the sample set included a blind duplicate used to assess method reproducibility. All data were subjected to statistical analysis with outliers eliminated from the data set. Results The percent coefficient of variation (%CV) of the sample set ranged from approximately 7 to 15%. Conclusions Further experimental details are described in the body of the manuscript and the results indicate the method is suitable for the determination of (±)-catechin and (±)-epicatechin in cocoa and chocolate products and represents the first collaborative study of this HPLC method for these compounds in these matrices. PMID:21729298

  10. Improved Quantitation of Gluten in Wheat Starch for Celiac Disease Patients by Gel-Permeation High-Performance Liquid Chromatography with Fluorescence Detection (GP-HPLC-FLD).

    Science.gov (United States)

    Scherf, Katharina Anne; Wieser, Herbert; Koehler, Peter

    2016-10-12

    Purified wheat starch (WSt) is commonly used in gluten-free products for celiac disease (CD) patients. It is mostly well-tolerated, but doubts about its safety for CD patients persist. One reason may be that most ELISA kits primarily recognize the alcohol-soluble gliadin fraction of gluten, but insufficiently target the alcohol-insoluble glutenin fraction. To address this problem, a new sensitive method based on the sequential extraction of gliadins, glutenins, and gluten from WSt followed by gel-permeation high-performance liquid chromatography with fluorescence detection (GP-HPLC-FLD) was developed. It revealed that considerable amounts of glutenins were present in most WSt. The gluten contents quantitated by GP-HPLC-FLD as sum of gliadins and glutenins were higher than those by R5 ELISA (gluten as gliadin content multiplied by a factor of 2) in 19 out of 26 WSt. Despite its limited selectivity, GP-HPLC-FLD may be applied as confirmatory method to ELISA to quantitate gluten in WSt.

  11. Determination of Tinopal CBS-X in rice papers and rice noodles using HPLC with fluorescence detection and LC-MS/MS.

    Science.gov (United States)

    Ko, Kyung Yuk; Lee, Chae A; Choi, Jae Chon; Kim, Meehye

    2014-01-01

    To date there have been no reports of methods to determine Tinopal CBS-X. We developed a rapid and simple method to determine the Tinopal CBS-X content in rice noodles and rice papers using HPLC equipped with fluorescence detection. Heating the rice noodles and rice papers to 80°C after adding 75% methanol solution induced the release of Tinopal CBS-X from processed rice products. Tinopal CBS-X was separated using an isocratic mobile phase comprising 50% acetonitrile/water containing 0.4% tetrabutyl ammonium hydrogen sulphate at pH 8.0. The samples suspected to be positive by HPLC analysis were then confirmed by LC-MS/MS analysis. This study also investigated the Tinopal CBS-X content of three rice noodle products and two rice papers. The limits of quantification for rice papers and rice noodles were 1.58 and 1.51 µg kg(-1), respectively, and their correlation curves showed good linearity with r(2) ≥ 0.9997 and ≥ 0.9998, respectively. Moreover, rice papers had recoveries of 70.3-83.3% with precision ranging from 5.0% to 7.9%, whereas rice noodles had slightly lower recoveries of 63.4-78.7% and precisions of 8.5-11.5%. Only one rice noodle product contained Tinopal CBS-X, at around 2.1 mg kg(-1), whereas it was not detected in four other samples. Consequently, Tinopal CBS-X from rice noodles and rice papers can be successfully detected using the developed pre-treatment and ion-pairing HPLC system coupled with fluorescence detection.

  12. Fourth-derivative synchronous spectrofluorimetry and HPLC with fluorescence detection as two analytical techniques for the simultaneous determination of itopride and domperidone.

    Science.gov (United States)

    Ibrahim, Fawzia; Nasr, Jenny Jeehan

    2016-02-01

    Two simple, rapid and sensitive methods, namely, fourth-derivative synchronous spectrofluorimetry (method I) and HPLC with fluorescence detection (method II) were developed for the simultaneous analysis of a binary mixture of itopride HCl (ITP) and domperidone (DOM) without prior separation. The first method was based on measuring the fourth derivative of the synchronous fluorescence spectra of the two drugs at Δλ = 40 nm in methanol. The different experimental parameters affecting the synchronous fluorescence of the studied drugs were carefully optimized. Chromatographic separation was performed in fluorescence detection at 344 nm after excitation at 285 nm. A mobile phase composed of a mixture of 0.02 M phosphate buffer with acetonitrile in a ratio of 55 : 45, pH 4.5, was used at a flow rate of 1 mL/min. Linearity ranges were found to be 0.1-2 µg/mL for ITP in both methods, whereas those for DOM were found to be 0.08-2 and 0.05-1.5 µg/mL in methods I and II, respectively. The proposed methods were successfully applied for the determination of the studied drugs in synthetic mixtures and laboratory-prepared tablets.

  13. Enantiomeric separation of D,L-tryptophan and D,L-kynurenine by HPLC using pre-column fluorescence derivatization with R(-)-DBD-PyNCS.

    Science.gov (United States)

    Iizuka, Hideaki; Hirasa, Yasushi; Kubo, Kazumi; Ishii, Kana; Toyo'oka, Toshimasa; Fukushima, Takeshi

    2011-07-01

    The enantiomeric separation of D,L-tryptophan (Trp) and D,L-kynurenine (KYN) was investigated by high-performance liquid chromatography using pre-column fluorescence derivatization with a chiral fluorescent labeling reagent, R(-)-4-(3-isothiocyanatopyrrolidin-1-yl)-7- (N,N-dimethylaminosulfonyl)-2,1,3-benzoxadiazole [R(-)-DBD-PyNCS]. Using an octadecylsilica column, namely, an Inertsil ODS-3 column (250 x 2.0 mm; i.d., 3 μm), four fluorescence peaks of D- and L-Trp as well as D- and L-KYN derivatized with R(-)-DBD-PyNCS were clearly observed, and their chemical structures were confirmed by HPLC-time-of-flight-mass spectrometry. Simultaneous separation was achieved under the mobile phase condition of 1.5% acetic acid in H₂O-CH₃CN (60:40), and the separation factors of D,L-Trp and D,L-KYN derivatized with R(-)-DBD-PyNCS were 1.22 and 1.19, respectively. Fluorescence detection was carried out by setting the emission wavelength at 565 nm, and the excitation wavelength at 440 nm, and the detection limits were approximately 0.3-0.5 pmol (signal-to-noise ratio of 3).

  14. Simultaneous determination of three banned psychiatric drugs in pig feed and tissue using solid-phase reactor on-line oxidizing and HPLC-fluorescence detection.

    Science.gov (United States)

    Qi, Liang; Duan, Li-Min; Sun, Xiao-Huan; Zhang, Jing; Zhang, Zhi-Qi

    2015-10-01

    The banned addition of psychiatric drugs such as phenothiazines to animal feed and foodstuffs increases the risk of human organ lesion. Phenothiazines usually exhibit weak native fluorescence and can be oxidized to strongly fluorescent compounds. In this study, a novel, sensitive and convenient method of HPLC-fluorescence detection based on post-column on-line oxidizing with lead dioxide solid-phase reactor has been developed for simultaneous determination of three banned psychotropic drugs, promethazine, chlorpromazine and thioridazine. Three compounds were successfully separated on an Agilent TC-C18 column with mobile phase of acetonitrile (A) and water (B), both containing 0.5% (v/v) formic acid. A gradient elution was programmed and fluorimetric detection was performed at λex /λem of 332/373 nm for promethazine, 340/380 nm for chlorpromazine and 352/432 nm for thioridazine. The calibration graphs gave good linearity over the concentration ranges of 30.0-4976.4 µg/L for promethazine, 2.0-2153.2 µg/L for chlorpromazine, and 15.0-3088.0 µg/L for thioridazine, and correlation coefficients (r) were ≥0.995. The method was applied to the determination of phenothiazines in pig feed and pig tissue, and the average spiked recoveries were in the range 69.1-115.4%.

  15. HPLC/FLD测定聚酯纤维纺织品中9种非离子荧光增白剂%Determination of nine non-ionic fluorescent whitening agent residues in polyester fibre textile by HPLC/FLD

    Institute of Scientific and Technical Information of China (English)

    吴晓琼; 丁友超; 汤娟; 徐海; 郑丹丹

    2016-01-01

    建立了纺织品中DT、SWN、OB、KSN、ER-Ⅰ、ER-Ⅱ、KCB、OB-Ⅰ、OB-Ⅱ9种非离子荧光增白剂(FWAs)的前处理方法以及高效色谱荧光检测(HPLC/FLD)方法。样品经二甲苯提取、浓缩、定容后,进行HPLC/FLD定性定量分析。采用Phenomenex Gemini 5u C18(150 mm×4.6 mm,5μm)色谱柱进行分离,以乙腈-2.5 mmol/L乙酸铵溶液为流动相,梯度洗脱。9种非离子FWAs的相关系数均不小于0.9993,定量下限(LOQs,S/N=10)为0.70~0.95 mg/kg。不同添加水平下,9种非离子FWAs的回收率为83.0%~100%,相对标准偏差(RSDs)为4.0%~8.8%。该方法简单、灵敏度高,具有一定的实际应用价值。%A sample pertreatment coupled to high performance with fluorescent detector (HPLC/FLD) method was developed for determination of nine non-ionic fluorescent whitening agents, including 1,2-bis(5-methyl-2-benzoxazole)ethylene (DT), 7-diethylamino-4-methylcoumarin (SWN), 2,2′-(2,5-thiophenedi⁃yl)bis[5-(1,1-dimethylethyl)-benzoxazol] (OB), 2-[4-[2-[4-(2-benzoxazolyl)phenyl]ethenyl]phenyl]-5-methyl-benzoxazol (KSN), 1,4-bis(2-cyano styryl)benzene (ER-Ⅰ), 1-(2-cyano styryl)-4-1-(2-cyano styryl)-4-(4-cy⁃ano styryl) benzene (ER-Ⅱ), 2,2′-(1,4-naphthalenediyl)bis-benzoxazol (KCB), 4,4′-bis[2-(2-methoxyphenyl) ethenyl]-1′-biphenyl (OB-Ⅰ), 2,2′-(1,2-ethenediyldi-4,1-phenylene)bis(5-methyl-benzoxazol) (OB-Ⅱ) in textile. The sample was extracted with xylene and concentrated, and then qualitatively and quantitatively ana⁃lyzed by HPLC/FLD. The separation of target compounds was performed on a phenomenex Gemini 5u C18 (150 mm × 4.6 mm,5 μm) column by gradient elution using acetonitrile/ammonium acetate solution (containing 2.5 mmol/L ammonium acetate) as mobile phase. The correlation coefficients of nine kinds of non-ionic fluo⁃rescent whitening agents was not less than 0.999 3. The limits of quantification of the nine compounds (LOQs, S/N=10) were 0

  16. Determination of the major constituents in fruit of Arctium lappa L. by matrix solid-phase dispersion extraction coupled with HPLC separation and fluorescence detection.

    Science.gov (United States)

    Liu, He; Zhang, Yupu; Sun, Yantao; Wang, Xue; Zhai, Yujuan; Sun, Ye; Sun, Shuo; Yu, Aimin; Zhang, Hanqi; Wang, Yinghua

    2010-10-15

    The arctiin and arctigenin in the fruit of Arctium lappa L. were extracted by matrix solid-phase dispersion (MSPD) and determined by high-performance liquid chromatography (HPLC) with fluorescence detection. The experimental conditions for the MSPD were optimized. Silica gel was selected as dispersion adsorbent and methanol as elution solvent. The calibration curve showed good relationship (r>0.9998) in the concentration range of 0.010-5.0μgmL(-1) for arctiin and 0.025-7.5μgmL(-1) for arctigenin. The recoveries were between 74.4% and 100%. The proposed method consumed less sample, time and solvent compared with conventional methods, including ultrasonic and Soxhlet extraction.

  17. Determination of Gastrodin in Gastrodia elata Blume by HPLC with ELSD and DAD%高效液相色谱法分离/蒸发光散射和紫外检测法测定天麻中天麻甙含量

    Institute of Scientific and Technical Information of China (English)

    魏泱; 丁明玉; 李红霞

    2001-01-01

    A normal-phase high performance chromatographic method with evaporative light-scattering detector(ELSD) and diode array detector(DAD), using n-hexane/methanol/ethyl acetate(volume ratio 6∶3∶2) as the mobile phase, was established for the direct determination of active constituents-gastrodin(GA) in the ethanol extracts of Gastrodia elata Blume. An silica gel column Zorbax RX-SIL[46 mm (i.d.)×25 cm, 5 μm] was employed. Regression equations revealed the linear relationships(r=0.998 and 0.999) between the peak areas of GA measured by ELSD and DAD and its concentration. The recoveries of GA measured by ELSD and DAD are 104.22% and 97.65%, respectively. The contents of GA measured by ELSD and DAD are 13.9 mg/g(2.79% RSD) and 12.1 mg/g(2.99% RSD) respectively in Gastrodia elata Blume. The detection limits of ELSD and DAD(S/N=3) are 3 mg/L and 1 mg/L respectively. This method is simple, selective and sensitive.

  18. HPLC - DAD analysis of lycorine in Amaryllidaceae species.

    Science.gov (United States)

    Kaya, Gulen Irem; Cicek, Derya; Sarikaya, Buket; Onur, Mustafa Ali; Somer, Nehir Unver

    2010-06-01

    Lycorine, the most frequent alkaloid found in Amaryllidaceae plants, has been proven to have various biological activities. Therefore, it is important to quantify this compound in Amaryllidacaeae species. In this study, a reversed-phase high-performance liquid chromatographic method has been used and validated for the determination of lycorine in Amaryllidaceae plants. A simple method for the extraction of lycorine in low-mass plant samples was employed utilizing columns pre-packed with diatomaceous earth (Extrelut). This method was applied to the aerial parts and bulbs of Sternbergia sicula Tineo ex Guss., S. lutea (L.) Ker-Gawl. ex Sprengel and Pancratium maritimum L. (Amaryllidaceae) collected during two different vegetation periods. The chromatographic separation was performed using an isocratic system with a mobile phase of trifluoroacetic acid-water-acetonitrile (0.01:90:10) applied at a flow rate 1 mL min(-1) using diode array detector. Validation procedures showed that the method was specific, accurate and precise. The amount of lycorine in S. sicula samples ranged between 0.10-0.53%, whereas in those of S. lutea and P. maritimum, the range was 0.19-0.40% and 0.05-0.14%, respectively.

  19. Forensic intoxication with clobazam: HPLC/DAD/MSD analysis

    OpenAIRE

    2004-01-01

    Clobazam (Castillium®, Urbanil®), a benzodiazepine often used as an anxiolytic and in the treatment of epilepsy, is considered a relatively safe drug. The authors present a fatal case with a 49-year-old female, found dead at home. She had been undergoing psychiatric treatment and was a chronic alcoholic. The autopsy findings were unremarkable, except for multivisceral congestion, steatosis and a small piece of a plastic blister pack in the stomach. Bronchopneumonia, bronchitis and bronchiolit...

  20. Determination of Related Substances in Cisatracurium Besilate by HPLC-DAD%高效液相色谱-二极管阵列检测器法测定苯磺顺阿曲库铵的有关物质

    Institute of Scientific and Technical Information of China (English)

    杜云; 王磊; 顾维

    2015-01-01

    目的:建立测定苯磺顺阿曲库铵中有关物质的高效液相色谱法。方法采用高效液相色谱-二极管阵列检测器法。流动相梯度洗脱,流动相A为1.02%磷酸二氢钾缓冲溶液(用磷酸调节pH至3.1)-甲醇-乙腈(75:5:20),流动相B为1.02%磷酸二氢钾缓冲溶液(用磷酸调节pH至3.1)-甲醇-乙腈(50:30:20);色谱柱为Thermo Syncronis C18柱(250 mm ×4.6 mm,5μm),流量为1.0 mL/min,柱温为30℃,进样体积为20μL,检测波长为280 nm。结果苯磺顺阿曲库铵质量浓度线性范围为0.005~1.0 g/L( r=0.9999),检测限为5 ng/mL( S/ N≥3);杂质E、杂质G、中间体Ⅻ和中间体ⅩⅢ的质量浓度线性范围分别为0.8~40.0μg/mL,1.2~60.0μg/mL,40~200μg/mL,1.6~80.0μg/mL( r≥0.9998),检测限分别为40 ng/mL,30 ng/mL,100 ng/mL,80 ng/mL( S/ N≥3)。结论该法检测苯磺顺阿曲库铵的有关物质,灵敏度高、专属性强,结果准确可靠。%Objective To establish a HPLC method for the determination of related substances in cisatracurium besilate. Methods HPLC-diode array detector(DAD)was adopted. The mobile phase was performed the gradient elution. The mobile phase A was 1. 02%potassium dihydrogen phosphate buffer solution (adjusting to pH 3. 1 with phosphoric acid)-methanol-acetontrile (75 :5 :20) and the mobile phase B was 1. 02% potassium dihydrogen phosphate buffer solution(adjusting to pH 3. 1 with phosphoric acid)-methanol-acetontrile(50 :30 :20). The separation of analyte was performed on a Thermo Syncronis C18 column(250 mm × 4. 6 mm,5 μm). The flow rate was 1. 0 mL/min,the column temperature was set at 30 ℃ and the injection volume was 20 μL. DAD was set at 280 nm. Results The linear range of cisatracurium besilate was 0. 005-1. 0 g/L( r=0. 999 9),the minimum detection limit was 5 ng/mL( S/N≥3);the linear ranges of mass concentrations for impurity E and G,midbody Ⅻ and

  1. Simple and sensitive HPLC method with fluorescence detection for the measurement of ibuprofen in rat plasma: application to a long-lasting dosage form.

    Science.gov (United States)

    Hassan, Ahmed Sheikh; Sapin, Anne; Ubrich, Nathalie; Maincent, Philippe; Bolzan, Claire; Leroy, Pierre

    2008-10-01

    A simple and sensitive high-performance liquid chromatography (HPLC) assay applied to the measurement of ibuprofen in rat plasma has been developed. Two parameters have been investigated to improve ibuprofen detectability using fluorescence detection: variation of mobile phase pH and the use of beta-cyclodextrin (beta-CD). Increasing the pH value from 2.5 to 6.5 and adding 5 mM beta-CD enhanced the fluorescence signal (lambda(exc) = 224 nm; lambda(em) = 290 nm) by 2.5 and 1.3-fold, respectively, when using standards. In the case of plasma samples, only pH variation significantly lowered detection and quantification limits, down to 10 and 35 ng/mL, respectively. Full selectivity was obtained with a single step for plasma treatment, that is, protein precipitation with acidified acetonitrile. The validated method was applied to a pharmacokinetic study of ibuprofen encapsulated in microspheres and subcutaneously administered to rats.

  2. Separation and quantitative determination of cinacalcet metabolites in urine sample using RP-HPLC after derivation with a fluorescent labeling reagent.

    Science.gov (United States)

    Farnoudian-Habibi, Amir; Jaymand, Mehdi

    2016-08-01

    In this investigation, a novel strategy for separation and quantitative determination of four metabolites of cinacalcet (M2a-Glu, M2b-Glu, M7-Gly, and M8-Gly) in human urine is suggested. The analytical assay is based on a pre-column derivation procedure of cinacalcet metabolites with 1-pyrenyldiazomethane (PDAM) as a fluorescent labeling reagent, and subsequently separation and quantitative determination with reverse-phase high-performance liquid chromatography (RP-HPLC) coupled with a fluorescence detector. Metabolites were separated on a Microsorb-MV 100-5 C18 chromatography column (250×4.6mm, 5μm) using acetate buffer (pH 3.5):methanol (30:70 v/v) as mobile phase at a flow rate of 1.0mLmin(-1). The method was fully validated in terms of linearity (r(2)>0.996; 1-10ngmL(-1)), precision (both intra-day and inter-day; RSD<6.2%), accuracy (92-110%), specificity, robustness (0.15%

  3. Dad's Playbook: Coaching Kids to Read

    Science.gov (United States)

    National Institute for Literacy, 2006

    2006-01-01

    By taking the time to read to and with their children, fathers can play an important role in helping children learn to read. "Dad's Playbook" tells the stories of 20 dads from different walks of life who are giving their kids the best shot at a bright future by helping them learn to read. This National Institute for Literacy Partnership for…

  4. Determination of etimicin in rat plasma using 9-fluorenylmethyl chloroformate precolumn derivatization by HPLC with fluorescence detection

    OpenAIRE

    Chang,Xiaojuan; Yu, Zhengping

    2011-01-01

    A new and sensitive method was developed for the determination of the etimicin (ETM) using precolumn derivatization (PCD) with 9-fluorenylmethyl chloroformate (FMOC-Cl) by a reversed phase separation and subsequent fluorescence detection. The PCD conditions were fully optimized towards the lowest limit of detection. The chromatographic separation was carried out on an Agilent XDB-C8 column at 25 ºC using a constant flow rate of 1.0 mL min-1 and mobile phase of acetonitrile/water (87:13, v/v)....

  5. Fluorescence detection of Zabofloxacin, a novel fluoroquinolone antibiotic, in plasma, bile, and urine by HPLC: the first oral and intravenous applications in a pharmacokinetic study in rats.

    Science.gov (United States)

    Jin, Hyo Eon; Kang, In Hyul; Shim, Chang Koo

    2011-01-01

    To develop an HPLC method using fluorescence detection for the pharmacokinetic evaluation of levels of zabofloxacin, a novel broad spectrum fluoroquinolone antibiotic, in the plasma, bile and urine of rats. A simple reversed-phase HPLC method using a C18 column with fluorescence detection was developed and validated for the simultaneous determination of zabofloxain and enrofloxacin as an internal standard. The plasma sample was treated with methanol for protein precipitation, and treatment of the bile and urine samples included deproteinization and extraction using chloroform. The applicability of the developed assay method to pharmacokinetic studies of zabofloxacin in rats was examined. Zabofloxacin was intravenously and orally administered to rats at a dose of 20 mg/kg. The limits of quantification (LOQ) was determined to be 50 ng/mL for the plasma with acceptable linearity ranging from 50 to 25,000 ng/mL (R>0.999), and 0.5 μg/mL for the bile and urine samples with acceptable linearity ranging from 0.5 to 100 μg/mL (R>0.999). The validation parameters for zabofloxacin were found to be acceptable according to FDA assay validation (2001). While zabofloxacin in plasma and urine has been stable in all tested handling conditions, it has been unstable in bile during freeze-thaw cycles for 24 h at room temperature. Following intravenous and oral administration of zabofloxacin to rats at a dose of 20 mg/kg, concentration was quantifiable in plasma for up to 8 h. The bioavailability of zabofloxacin was 27.7%, and it was excreted into bile and urine at about 8% each per oral administration. These observations suggest that a validated assay can be used in pharmacokinetic studies of zabofloxacin in small animals. Due to the limited stability of zabofloxcin in rat bile, freeze-thaw cycles or prolonged handling at room temperature is not recommended. This article is open to POST-PUBLICATION REVIEW. Registered readers (see "For Readers") may comment by clicking on ABSTRACT on

  6. 高效液相色谱分离-在线光化学衍生/荧光光谱法测定5种水溶性维生素的研究%Simultaneous Determination of 5 Water-soluble Vitamins by HPLC with On-line Photochemical Derivatization and Fluorescence Detection

    Institute of Scientific and Technical Information of China (English)

    刘珺; 方芳; 陈婷; 段华玲; 弓振斌

    2011-01-01

    建立了高效液相色谱分离-在线光化学衍生/荧光光谱法测定水溶性维生素烟酸(NIA)、烟酸胺(NIC)、B1、B12及B2的新方法.以含有0.018 mol/L三乙胺、0.002 mol/L庚烷磺酸钠的0.05 mol/L磷酸盐缓冲液(A相,pH 5.8)和甲醇(B相)为流动相(85:15),等度洗脱分离5种水溶性维生素;将自制的程序控制时间/光强光化学反应器接在二极管阵列检测器(DAD)和荧光检测器(FLD)之间,进行光化学衍生.优化了光化学衍生的条件,并用于复合维生素片中水溶性维生素的测定.在优化实验条件下,5种水溶性维生素的线性范围分别为:NIA,NIC为0.1-10.0 mg/L,B1、B12为0.2-20.0 mg/L,B2为0.01-1.0 mg/L,方法的检出限(LOD,3σ)和定量下限(LOQ,10σ)分别为1.5-25.4 μg/和5.0-84.7μg/L,优于传统的高效液相色谱分离/二极管阵列检测方法.方法的相对标准偏差(n=7)不大于7.8%,样品的加标回收率为98%-103%.%A high performance liquid chromatographic ( HPLC ) method using on-line photochemical derivatization and fluorescence detection was developed for the simultaneous determination of nicotinic acid(NIA), nicotinamide( NIC), thiamine hydrochloride ( B1), riboflavin ( B2) and cyanocobala-min(B12). A self-built time/energy programmed photochemical reactor (PCR) with an ultraviolet mercury lamp was installed between the diode-array detector (DAD) and fluorescence detector( FLD) to convert the non- or weakly fluorescent vitamins into fluorescence compounds. The separation of NIA, NIC, B,, B12 and B2 was achieved within 12 min using 0.05 mol/L phosphate buffer(pH 5. 8) -0. 018 mol/L triethylamine -0. 002 mol/L sodium heptane sulfonate( A) and methanol(B) as mobile phase by isocratic elution( A : B, 85 : 15). The separation and photochemical derivatization conditions were optimized. Under the optimal conditions, the calibration curves of five vitamins were linear in the range of 0. 1 - 10. 0 mg/L for NIA and NIC, 0. 2 - 20. 0 mg/L for B, and B12

  7. HPLC determination of ibuprofen, diclofenac and salicylic acid using hollow fiber-based liquid phase microextraction (HF-LPME).

    Science.gov (United States)

    Ramos Payán, María; Bello López, Miguel Angel; Fernández-Torres, Rut; Pérez Bernal, Juan Luis; Callejón Mochón, Manuel

    2009-10-27

    This paper describes an extraction method using a polypropylene membrane supporting dihexyl ether (three-phase hollow fiber-based liquid phase microextraction (HF-LPME)) for the analysis of several pharmaceuticals (salicylic acid (SAC), ibuprofen (IBU) and diclofenac (DIC)) followed by a HPLC determination using a monolithic silica type HPLC column, that allows lower retention times than the usual packed columns with adequate resolution. Detection was realized by means of a coupled in series diode array (DAD) and fluorescence (FLD) detectors. HF-LPME is a relatively new technique employed in analytical chemistry for sample pretreatment which offers more selectivity and sensitivity than any traditional extraction technique. Detection limits by DAD are 12, 53 and 40 ng mL(-1) for salicylic acid, diclofenac and ibuprofen, respectively and by FLD 7 and 2 ng mL(-1) for salicylic acid, and ibuprofen. The method has been successfully applied to their direct determination in human urine and the results obtained demonstrated that could be also applied to the determination of the corresponding metabolites.

  8. Determination of the Component of Fluorescent Whitening Agent CBS-127 by HPLC%HPLC法测定荧光增白剂CBS-127的含量

    Institute of Scientific and Technical Information of China (English)

    李自红; 魏悦; 范毅; 董建军

    2013-01-01

    The method for determinating the content of Fluorescent Whitening Agent CBS-127 by HPLC was established.Sunfire C18 column (4.6 mm×150 mm,5 μm) was used with methanol-water (90%:10%) as the mobile phase.The flow rate of the mobile phase was 1.0 mL/min.The detection wavelength was 356 nm.The column temperature was 40 C.There was good linear relationship in the range of 0.5~2.5 μg (r=0.999 9).The recovery rate was 99.78% and RSD value is 0.7% (n=6).This method is high accuracy,good reproducibility,reliable results,which is an effective method for the Fluorescent Whitening Agent CBS-127.%建立了高效液相色谱法测定荧光增白剂CBS-127含量的方法.采用Sunfire C18 (4.6 mm×150 mm,5μm)色谱柱,流动相为甲醇-水(90%:10%),流速为1.0 mL/min,紫外检测波长为356 nm,柱温40℃.CBS-127在0.5~2.5 μg范围内有良好线性关系(r=0.999 9);加样回收率为99.78%,RSD值为0.7%(n=6).该方法精密度高、重现性好、结果准确可靠,为荧光增白剂CBS-127的质量评价提供了有效的方法.

  9. Comparison of different sample treatments for the analysis of ochratoxin A in wine by capillary HPLC with laser-induced fluorescence detection.

    Science.gov (United States)

    Arroyo-Manzanares, Natalia; García-Campaña, Ana M; Gámiz-Gracia, Laura

    2011-11-01

    Ochratoxin A (OTA) is a mycotoxin naturally found in various foods, including wine. As OTA is considered as a possible human carcinogen, the maximum concentration for this compound has been established at 2 μg kg(-1) in wine by the EU (Directive (CE) No 1881/2006). Typically, immunoaffinity columns have been used for its extraction. However, simpler, more efficient and less contaminant extraction systems are demanding. In this work, dispersive liquid-liquid microextraction using ionic liquid as extractant solvent (IL-DLLME) and the QuEChERS procedure, have been evaluated and compared for extraction of OTA in wine samples. Laser-induced fluorescence (LIF, He-Cd Laser excitation at 325 nm) coupled with capillary HPLC has been used for the determination of OTA, using a sodium dodecyl sulfate micellar solution in the mobile phase to increase the fluorescence intensity. Matrix-matched calibration curves were established for both methods, obtaining LODs (3× S/N) of 5.2 ng·L(-1) and 85.7 ng·L(-1) for IL-DLLME and QuEChERS, respectively. Clean extracts were obtained for white, rose and red wines with both methods, with recoveries between 88.7-94.2% for IL-DLLME and between 82.6-86.2% for QuEChERS. The precision was evaluated in terms of repeatability (n = 9) and intermediate precision (n = 15), being ≤ 8.5% for IL-DLLME and ≤ 5.4% for QuEChERS.

  10. Design of a method for generation of gas-phase hydroxyl radicals, and use of HPLC with fluorescence detection to assess the antioxidant capacity of natural essential oils.

    Science.gov (United States)

    Pezo, D; Salafranca, J; Nerín, C

    2006-08-01

    The use of natural antioxidants is of increasing importance in the human diet, because they are recognised as compounds essential to health which minimize or delay the aging process. Despite apparent simplicity, however, it is very difficult to measure and quantify such properties, for which a robust analytical method is required. Because oxidation usually is caused by the presence of OH* radicals, a new method involving the in-situ, vapour-phase generation of these radicals and their quantification in the presence and absence of potential antioxidant extracts has been developed. The oxidant atmosphere generated from hydrogen peroxide is carried by an air stream through an empty quartz chamber in which UV radiation promotes the formation of radicals by a photochemical reaction. The products then pass through a cartridge containing the essential oil, finally bubbling into an impinger containing an aqueous solution of salicylic acid, at pH 4.5, which reacts with the OH* radicals forming 2,5-dihydroxybenzoic acid. This solution is quantified by RP-HPLC using UV and fluorescence detectors connected in series. Detection and quantification limits for OH* radicals were approximately 0.01 pg g(-1) air. Description and optimization of the method are discussed, as also is the antioxidant performance of an extract of ginger (Zingiber officinale R.), which reduced the oxidation process by up to 92%.

  11. Development and validation of a rapid HPLC- fluorescence method for simultaneous determination of venlafaxine and its major metabolites in human plasma

    Directory of Open Access Journals (Sweden)

    Y.H Ardakani

    2010-06-01

    Full Text Available "n Background and the purpose of the study:To develop a simple, rapid and accurate HPLC method for the measurement of the venlafaxine and its main metabolites, O-desmethylvenlafaxine and O,N-didesmethylvenlafaxine in pharmacokinetic studies and therapeutic drug monitoring.Method: Chromatographic separation was achieved with a ChromolithTM Performance RP-18e 100 mm×4.6 mm column equipped with a Fluorescence detectore (λex 200 nm/λem 300 nm The mobile phase of methanol:water (35:65, v/v adjusted to pH 2.5 by phosphoric acid was passed through the column in an isocratic mode at flow rate of 2 ml/min. The sample preparation involved a simple, one-step, extraction with ethyl acetate. "nResults:The calibration curves were linear in the concentration range of 1-300 ng/ml for all analytes (r2 > 0.998. The lower limit of quantification was 1 ng/ml for all analytes. Within and between day precisions in the measurement of quality control (QC of samples were in the range of 1.8-14.1% for all analytes. Conclusion:The developed procedure was used to assess the pharmacokinetics of venlafaxine and its main metabolites following oral administration of 75 mg venlafaxine to a healthy subject.

  12. Simultaneous determination of homocysteine, methionine and cysteine in maternal plasma after delivery by HPLC-fluorescence detection with DBD-F as a label.

    Science.gov (United States)

    Wada, Mitsuhiro; Hirose, Maki; Kuroki, Mana; Ikeda, Rie; Sekitani, Yui; Takamura, Noboru; Kuroda, Naotaka; Nakashima, Kenichiro

    2013-06-01

    An HPLC-fluorescence (FL) method for determination of sulfur-containing amino acids such as homocysteine (Hcy), methionine (Met) and cysteine (Cys) in human plasma was developed. The sulfur-containing amino acids were labeled with 4-(N,N-dimethylaminosulfonyl)-7-fluoro-2,1,3-benzoxadiazole (DBD-F). Calibration curves in the range of 1-100 µm (Hcy and Met) and 5-500 µm (Cys) indicated good linearities (r ≥ 0.998). The limits of detection at a signal-to-noise ratio of 3 were 0.13 (Hcy), 0.02 (Met) and 0.11 µm (Cys), respectively. Acceptable results for accuracy and precision of intra- and inter-day measurements were obtained. The results of Hcy and Cys obtained by the proposed method indicated good correlations with the conventional method (r > 0.911, n = 20). Furthermore, the method was applied to determination of the sulfur-containing amino acids in maternal plasma (n = 200) after delivery. The concentrations of Hcy, Met and Cys as a median (inter quartile range, Q1 and Q3 ) were 5.37 (3.32-7.79) μm, 25.20 (20.10-31.06) μm and 147.25 (102.81-189.31) μm, respectively.

  13. Simultaneous determination of selected eicosanoids by reversed-phase HPLC method using fluorescence detection and application to rat and human plasma, and rat heart and kidney samples.

    Science.gov (United States)

    Aghazadeh-Habashi, Ali; Asghar, Waheed; Jamali, Fakhreddin

    2015-06-10

    Eicosanoids are biologically active lipid-derived oxidative metabolites of arachidonic acid. We, herein, present an improved sensitive, selective and robust high performance liquid chromatography (HPLC)-fluorescence assay for simultaneous quantification of eicosanoids in human plasma and rat tissues. Aliquots of 200 μL of plasma or 30 mg of heart or kidney tissues were spiked with 16-hydroxydecanoic acid as internal standard, and extracted with anhydrous acetonitrile using solid phase cartridges. The eluted samples were dried, reconstituted in anhydrous acetonitrile and labeled with 2-(2,3-naphthalimino)ethyl-trifluoromethanesulphonate in the presence of saturated potassium fluoride solution in anhydrous acetonitrile and N,N-diiospropylethylamine as catalyst. The derivatized eicosanoids were extracted with anhydrous acetonitrile using solid phase cartridges. Chromatographic separation was achieved on a C18 reversed phase column using gradient mobile phase of 0.05% of formic acid:acetonitrile:water at 0.8 mL/min flow rate. The analytes were detected at excitation and emission wavelength of 260 and 396 nm, respectively. The assay was linear (r(2)≥ 0.98) in the concentration range of 0.01-2.5 μg/mL. The intra-day and inter-day coefficients variation was less than 19.8%. Using this assay, we were able to quantify arachidonic acid metabolites simultaneously in human and rat biological samples.

  14. Enantioselective determination of metoprolol and its metabolites in human urine high-performance liquid chromatography with fluorescence detection (HPLC-FLD) and tandem mass spectrometry (MS/MS).

    Science.gov (United States)

    Baranowska, Irena; Adolf, Weronika; Magiera, Sylwia

    2015-11-01

    A sensitive, stereoselective assay using solid phase extraction and high-performance liquid chromatography (HPLC) with fluorescence detection (FLD) was developed and validated for the analysis of enantiomers of metoprolol and its metabolites (α-hydroxymetoprolol, O-desmethylmetoprolol). Chiral separation was achieved using a CHIRALCEL OD-RH column, packed with cellulose tris-(3,5-dimethylphenyl-carbamate) stationary phase, employing a mobile phase composed by a mixture of 0.2% diethylamine in water and acetonitrile in gradient elution mode. Linear calibration curves were obtained over the range of 0.025-2.0μg/mL (R(2)>0.994) in urine for both enantiomers of metoprolol and its metabolites with quantitation limit of 0.025μg/mL. Intra and inter-day precision and accuracy were below 15% for both metoprolol and metabolites enantiomers. The recovery of enantiomer of metoprolol and its metabolite was greater than 68.0%, utilizing a SPE procedure. The method was tested with urine quality control samples and human urine fractions after administration of 50mg rac-metoprolol.

  15. Sensitive determination of trimetazidine in spiked human plasma by HPLC with fluorescence detection after pre-column derivatization with 9-fluorenylmethyl chloroformate.

    Science.gov (United States)

    Khedr, Alaa; Sheha, Mahmoud M; Darwish, Ibrahim A

    2007-09-01

    A high-performance liquid chromatographic method for the determination of trimetazidine dihydrochloride (TMZ) in spiked human plasma is described. The method is based on the pre-column derivatization with 9-fluorenylmethyl chloroformate (FMOC-Cl) using the fluorimetric detection technique. Fluoxetine HCl (FLX) was used as internal standard. Both, TMZ and FLX were completely derivatized after heating at 50 degrees C for 20 min in borate buffer pH 8.0. Samples were analyzed by high performance liquid chromatography (HPLC) using Zorbax-TMS column (250 mm x 4.6 mm, i.d., 5 microm) and mobile phase consist of acetonitrile, methanol and 20 mM sodium acetate pH 4.7 (44:6:50; v/v/v). Fluorescence detector (FLD) was adjusted at excitation and emission wavelengths; 265 and 311 nm, respectively. The linearity of the method was in the range of 4.5-200 ng/ml. Limits of detection (LOD) and quantification (LOQ) were 1.5 and 4.5 ng/ml, respectively. Trimetazidine recovery was 96.5+/-1.3% (n=6; RSD=2.1%).

  16. Low cost methodology for estrogens monitoring in water samples using dispersive liquid-liquid microextraction and HPLC with fluorescence detection.

    Science.gov (United States)

    Lima, Diana L D; Silva, Carla Patrícia; Otero, Marta; Esteves, Valdemar I

    2013-10-15

    A new low cost methodology for estrogens' analysis in water samples was developed in this work. Based on dispersive liquid-liquid microextraction followed by high-performance liquid chromatography with fluorescence detection, the developed method is fast, cheap, easy-to-use, uses low volumes of organic solvents and has the possibility of a large number of samples to be extracted in parallel. Under optimum conditions (sample volume: 8 mL; extraction solvent: 200 μL of chlorobenzene; dispersive solvent: 2000 μL of acetone), the enrichment factor and extraction recoveries were 145 and 72% for 17β-estradiol (E2) and 178 and 89% for 17α-ethinylestradiol (EE2), respectively. Limits of detection of 2.0 ng L(-1) for E2 and 6.5 ng L(-1) for EE2 were achieved, allowing the detection and quantification of these compounds in surface and waste water samples with concentrations from 12 to 32 ng L(-1) for E2 and from 11 to 18 ng L(-1) for EE2. Also, recovery tests were performed to evaluate possible matrix effects. Recoveries between 98% and 106% were obtained using humic acids (HA) to simulate the effect of organic matter, and between 86% and 120% in real water samples.

  17. HPLC-fluorescence detection method for determination of key intermediates of the lincomycin biosynthesis in fermentation broth.

    Science.gov (United States)

    Kameník, Zdenek; Kopecký, Jan; Marecková, Markéta; Ulanová, Dana; Novotná, Jitka; Pospísil, Stanislav; Olsovská, Jana

    2009-03-01

    The biosynthetic pathway of the clinically important antibiotic lincomycin is not known in details. The precise knowledge of the lincomycin biosynthesis is a prerequisite for generation of improved derivatives by means of combinatorial genetics. Methods allowing determination of the key intermediates are very important tools of the pathway investigation. Two new high-performance liquid chromatography methods with fluorescence detection for determination of lincomycin precursors in fermentation broth of Streptomyces lincolnensis and its lincomycin nonproducing mutants were developed. The first one enables simultaneous analysis of methylthiolincosamide (MTL) and N-demethyllincomycin (NDL), whereas the second one is suitable for 4-propyl-L-proline (PPL) assay. Both methods are based on the pre-column derivatization: MTL and NDL with 4-chloro-7-nitrobenzofurazan; PPL with o-phthaldialdehyde. The methods were validated with lower limit of quantification values of 2.50, 3.75, and 3.75 microg ml(-1) for MTL, NDL, and PPL, respectively. The inter- and intra-day accuracies and precisions were all within 12%. Stability of oxidized and derivatized analytes was investigated.

  18. Identification and Determination of Nicorandil and its Degradation Products by HPLC and GC/MS

    Institute of Scientific and Technical Information of China (English)

    Zhong Zhou CHENG; Ze Hui JIA; Yan CHEN; Li Ying CHEN; Hua LI

    2006-01-01

    A rapid and sensitive HPLC-DAD method is developed for simultaneous determination of nicorandil and its degradation products, N-(2-hydroxyethyl) nicotinamide, nitrate ion and nicotinic acid, using nicotinamide (NT) as internal standard, at wavelength 204 nm. Nicotinic acid is identified by HPLC and GC/MS. The method can also be applied to study kinetic of degradation processes of nicorandil in storage.

  19. Determination of 2,3-dimercaptosuccinic acid in mice blood and tissues by HPLC with fluorescence detection.

    Science.gov (United States)

    Ju, Xue Hai; Shi, Ying; Liu, Na; Guo, Rui Chen; Wang, Ben Jie; Cui, Xi

    2009-06-01

    2,3-Dimercaptosuccinic acid (DMSA) is an orally effective chelating agent for the treatment of heavy metal poisoning. The increasing therapeutic use of DMSA has stimulated the need for sensitive and selective methods for its determination in biological samples, as well as study on pharmacokinetics and tissue distribution. According to the previously reported method, an improved method was established for the determination of DMSA in mice blood and tissues, in which oxidized DMSA was reduced by the disulfide-reducing agent, dithiothreitol (DTT), and DMSA was converted to a highly fluorescent and stable derivative by reaction with monobromobimane (mBBr) in alkaline solution. Acetonitrile was used for deproteinization and dichloromethane was used for condensation and purification, which significantly shortened the amount of time used to process the sample. Meanwhile isocratical elution was performed and excellent separation of the DMSA derivative was obtained, this enabled a run finish within 20 min. The limits of quantitation were 0.025 microg/ml in brain and 0.1 microg/ml in blood, lung, heart, intestine, liver, spleen and kidney, respectively. The calibration curves were linear in all samples (r(2)>0.992) with a range of 0.025-1.6 microg/ml for brain homogenate and 0.1-6.4 microg/ml for blood and homogenates of lung, heart, intestine, liver, spleen and kidney, respectively. Therefore, the method is simple, rapid and sensitive, and it could be applicable to the studies in an animal model to evaluate the distribution of DMSA in blood and tissues.

  20. Simultaneous determination of sulpiride and mebeverine by HPLC method using fluorescence detection: application to real human plasma

    Directory of Open Access Journals (Sweden)

    Walash Mohamed I

    2012-02-01

    Full Text Available Abstract A new simple, rapid and sensitive reversed-phase liquid chromatographic method was developed and validated for the simultaneous determination of sulpiride (SUL and mebeverine Hydrochloride (MEB in the presence of their impurities and degradation products. The separation of these compounds was achieved within 6 min on a 250 mm, 4.6 mm i.d., 5 m particle size Waters®-C18 column using isocractic mobile phase containing a mixture of acetonitrile and 0.01 M dihydrogenphosphate buffer (45:55 at pH = 4.0. The analysis was performed at a flow rate of 1.0 mL/min with fluorescence-detection at excitation 300 nm and emission at 365 nm. The concentration-response relationship was linear over a concentration range of 10- 100 ng/mL for both MEB and SUL with a limit of detection 0.73 ng/mL and 0.85 ng/mL for MEB and SUL respectively. The proposed method was successfully applied for the analysis of both MEB and SUL in bulk with average recoveries of 100.22 ± 0.757% and 99.96 ± 0.625% respectively, and in commercial tablets with average recoveries of 100.04 ± 0.93% and 100.03 ± 0.376% for MEB and SUL respectively. The proposed method was successfully applied to the determination of MEB metabolite (veratic acid in real plasma simultaneously with SUL. The mean% recoveries (n = 3 for both MEB metabolite (veratic acid and SUL were 100.36 ± 2.92 and 99.06 ± 2.11 for spiked human plasma respectively. For real human plasma, the mean% recoveries (n = 3 were and respectively.

  1. Determination of closantel residues in milk and animal tissues by HPLC with fluorescence detection and SPE with oasis MAX cartridges.

    Science.gov (United States)

    Sun, Han-Wen; Wang, Feng-Chi; Ai, Lian-Feng

    2008-04-01

    A liquid chromatographic method for the determination of closantel residues in milk and tissues is developed and validated. An acetonitrile-acetone solution (80:20, v/v) is used for the extraction of closantel residues from milk and animal tissues, and the extract is purified by solid-phase extraction with Oasis MAX cartridges and a mixture of formic acid-acetonitrile (5:95, v/v) as the elution solution. A C(18) bonded silica column is used for chromatographic separation. The mobile phase consists of acetonitrile-water (85:15, v/v) containing 0.05% triethylamine at pH 2.5, adjusted with phosphoric acid with the flow-rate set at 1.0 mL/min. Using the fluorescence emission of closantel at lambda(ex) = 335 nm and lambda(ex) = 510 nm, the calibration curve is linear, with a correlation coefficient of 0.9999 over the concentration range of 10-5000 microg/kg for the tissue sample and 10-5000 microg/L for the milk sample. The detection limit (s/n = 3) is 3 microg/kg for tissue sample and 3 microg/L for milk sample. The intra- and inter-day repeatabilities are between 3.35-7.66% and 4.04-8.67%, respectively. The proposed method enables the quantitative determination of closantel residues at levels as low as 10 microg/kg in animal tissue samples and 10 microg/L in milk samples.

  2. A laser-induced fluorescence dual-fiber optic array detector applied to the rapid HPLC separation of polycyclic aromatic hydrocarbons

    Energy Technology Data Exchange (ETDEWEB)

    Hart, Sean J.; Hall, Gregory J.; Kenny, Jonathan E. [Tufts University, Chemistry Department, Medford, MA, (United States)

    2002-01-01

    A multi-channel detection system utilizing fiber optics has been developed for the laser-induced fluorescence (LIF) analysis of chromatographic eluents. It has been applied to the detection of polycyclic aromatic hydrocarbons (PAH) in a chromatographically overlapped standard mixture and to a complex soil sample extract obtained during fieldwork. The instrument utilizes dual-fiber optic arrays, one to deliver multiple excitation wavelengths (258-342 nm) generated by a Raman shifter, and the other to collect fluorescence generated by the sample at each excitation wavelength; the collected fluorescence is dispersed and detected with a spectrograph/CCD combination. The resulting data were arranged into excitation emission matrices (EEM) for visualization and data analysis. Rapid characterization of PAH mixtures was achieved under isocratic chromatographic conditions (1.5 mL min{sup -1} and 80% acetonitrile in water), with mid {mu}g L{sup -1} detection limits, in less than 4 minutes. The ability of the instrument to identify co-eluting compounds was demonstrated by identifying and quantifying analytes in the rapid analysis of a 17 component laboratory-prepared PAH mixture and a soil extracted sample. Identification and quantification were accomplished using rank annihilation factor analysis (RAFA) using pure component standards and the EEMs of mixtures measured during the rapid high-performance liquid chromatography (HPLC) method as the unknowns. The percentage errors of the retention times (RTs) determined using RAFA compared to the known RTs measured with a standard absorbance detector were between 0 and 11%. For the standard PAH mixture, all 17 components were identified correctly and for the soil extracted sample, all 8 analytes present were correctly identified with only one false positive. Overall, the system achieved excellent qualitative performance with semi-quantitative results in the concentration predictions of both the standard mixture and the real

  3. HPLC Quantification of Flavonoids and Biflavonoids in Cupressaceae Leaves

    OpenAIRE

    A. Romani; C. GALARDI; P. PINELLI; Mulinacci, N.; D. HEIMLER

    2002-01-01

    The aim of this investigation was to obtain qualitative and quantitative profiles of the flavonoid and biflavonoid composition of six cypress species - Cupressus funebris L., Cupressus semper- Wrens L., Cupressus glabra L., Cupressus arizonica L., Cupressus goveniana L., and Cupressus lusitanica L. HPLC-diode-array detection (DAD), HPLC-MS, and HPTLC were used to identify the individual compounds. A chromatographic method was optimized for identification and quantification of t...

  4. Kids With Two Moms or Two Dads

    Science.gov (United States)

    ... can be a mom and a dad, a single parent, a gay couple, or even someone who isn' ... With Parents Living With Stepparents Living With a Single Parent Talking to Your Parents Contact Us Print Resources ...

  5. Tissue Distribution Study of Dimercaptosuccinic Acid in Mice by HPLC-Fluorescence Method%HPLC-荧光法研究二巯基丁二酸在小鼠体内的组织分布特性

    Institute of Scientific and Technical Information of China (English)

    刘世军; 崔晞; 孙克明; 刘宪勇; 张敏; 王岩

    2011-01-01

    OBJECTIVE: To study the tissue distribution of dimercaptosuccinic acid (DMSA) in mice. METHODS: HPLC with fluorescence method was adopted. The biological samples were treated with sulfhedryl derivatization reagent. The determination was performed on AichromBond-AQ C18 colunm with mobile phase consisted of methanol-water (40:60, V/V, containing tetrabutyl ammonium bromide and sodium acetate) and flow rate of 1.0 mL·min-1. The column temperature was 30 ℃. The fluorescence detector equipped with 356 nm excitation and 450 nm emission filters was used for measurement of peaks. 36 healthy Kunming mice were randomly divided into 6 groups. One group was control group, the other 5 groups for the drug group. Drug group was given 2.6 mg·mL-1 DMSA 0.5 mL via i.g. gtt. The samples of blood, brain, heart, lungs, intestines, liver, spleen and kidney were collected and the DMSA was determined 0.5, 2, 4, 6, 12 h after administration. RESULTS: The good linear range of DMSA had been obtained (r>0.996). The lowest detection limit of brain and other samples were 0.025 μg·mL-1 and 0.1 μg·mL-1. The extraction recovery was more than 75% and the RSD of intra-day and inter-day were less than 15%. The peaks of drug concentration were reached at 0.5 h in intestine, liver, heart and blood, and at 2 h in kidney, lung, spleen and brain after an oral administration of DMSA. The concentration of DMSA in tissues was followed as: kidney>blood>intestine>lung>liver>spleen>heart>brain.CONCLUSIONS: The method is accurate, specific and reliable with less interference of endogenous substances, and it can be applied to the study of DMSA in mice tissue. After an oral administration, the DMSA is rapidly absorbed, distributed and eliminated.The tissue distribution of DMSA is excreted via kidney and liver.%目的:研究二巯基丁二酸(DMSA)在小鼠体内的组织分布特性.方法:采用高效液相色谱(HPLC)-荧光法.生物样品在加入巯基衍生化试剂mBBr后进行

  6. Simultaneous determination of four aflatoxins and ochratoxin A in ginger and related products by HPLC with fluorescence detection after immunoaffinity column clean-up and postcolumn photochemical derivatization.

    Science.gov (United States)

    Wen, Jing; Kong, Weijun; Wang, Jian; Yang, Meihua

    2013-12-01

    Ginger, a widely used spice and traditional Chinese medicine, is prone to be contaminated by mycotoxins. A simple, sensitive, and reproducible method based on immunoaffinity column clean-up coupled with HPLC and on-line postcolumn photochemical derivatization with fluorescence detection was developed for the simultaneous determination of aflatoxins (AFs) B1 , B2 , G1 , G2 , and ochratoxin A (OTA) in 25 batches of gingers and related products marketed in China for the first time. The samples were first extracted by ultrasonication with methanol/water (80:20, v/v) and then cleaned up with immunoaffinity columns for analysis. Under the optimized conditions, the LODs and LOQs for the five mycotoxins were 0.03-0.3 and 0.1-0.9 μg/kg, respectively. The average recoveries ranged from 81.3-100.8% for AFs and from 88.6-99.5% for OTA at three spiking levels. Good linearity was observed for the analytes with correlation coefficients all >0.9995. All moldy gingers were contaminated with at least one kind of the five investigated mycotoxins, while none of them were found in normal gingers. Ginger powder samples were contaminated slightly with the contamination levels below the LOQs, while ginger tea bags were mainly contaminated by OTA at 1.05-1.19 μg/kg and ginger black tea bags were mainly contaminated by AFs at 3.37-5.76 μg/kg. All the contamination levels were below the legally allowable limits.

  7. HPLC荧光检测法测定人血浆中的特拉唑嗪%Determination of terazosin in human plasma by HPLC with fluorescence detection

    Institute of Scientific and Technical Information of China (English)

    刘曼; 张丹; 王国才; 强桂芬; 杨漫; 张娅喃; 刘会臣

    2012-01-01

    OBJECTIVE To establish an HPLC method with fluorescence detection for determination of terazosin in human plasma. METHODS The separation was carried out on an Apollo C18 column using the mobile phase of acetonitrile and 50 mmol·L-1 potassium dihydrogen phosphate buffer solution(22 : 78)at a flow rate of 1. 1 mL·min-1. The column temperature was maintained at 30 ℃ and the injection volume was 10 μL. The excitation and emission wavelengths of the fluorescence detection were set at 237 nm and 370 run,respectively. The analyte and prazosin (internal standard) were extracted with diethylether - dichlormethane(3 :2 ) after alkalin-ized. RESULTS The linear range of terazosin in human plasma was 0. 5 -125.0 μg·L-1( r≥0. 9983). The intra-and inter - day RSD were both less than 10%. The mean extraction recovery was 88.0% ±4.4%. CONCLUSION The method is rapid, simple, sensitive and suitable for the clinical pharmacokinetic study of terazosin preparations.%目的 采用HPLC荧光检测法测定人血浆中的特拉唑嗪.方法 采用Apollo C18色谱柱(150 mm×4.6 mm,5μm),流动相为乙腈-50 mmol·L-1磷酸二氢钾缓冲液(22∶78),流速1.1 mL· min -1,柱温30℃,进样量10 μL,荧光检测器激发波长为238 nm、发射波长为370 nm.以哌唑嗪为内标,血浆样本碱化后经乙醚-二氯甲烷(3∶2)提取后进样.结果 特拉唑嗪的线性范围为0.5~125.0 μg·L-1(r≥0.9983);日内、日间RSD均小于10%;平均提取回收率为88.0%±4.4%.结论 所用方法简便、准确、灵敏,无杂质干扰,适用于临床上盐酸特拉唑嗪口服制剂的血药浓度监测及药动学研究.

  8. HPLC-荧光法和LC-ESI/MS法用于人血浆中替米沙坦分析的评价%Evaluation of HPLC -fluorescence and LC -ESI/MS for determination of telmisartan in human plasma

    Institute of Scientific and Technical Information of China (English)

    沈于兰; 冯芳; 武洁

    2006-01-01

    目的:建立替米沙坦血药浓度的HPLC-荧光检测法,并从方法学角度比较了该方法与HPLC-MS分析方法的异同.方法:血浆中加入内标米氮平后,醋酸乙酯提取,HPLC-荧光法分离、分析.色谱系统:汉邦ODS柱(150 mm×4.6 mm,5μm),甲醇-0.01 mol·L-1磷酸二氢钾水溶液(pH=6)(62:38,v/v)为流动相,流速1.0 mL·min-1,柱温35℃.LC-MS法用醋酸乙酯提取后,SIM检测.结果:2种方法中替米沙坦的线性范围均为0.5~400 ng·mL-1,HPLC-荧光法的最低检测限为0.5 ng·mL-1(取样量0.2 mL),HPLC-MS的最低检测浓度为0.1ng·mL-1(取样量0.5 mL).两者的提取回收率均≥80%.结论:通过对2种方法回收率的t检验发现,两法所得结果无显著性差异.HPLC-荧光法既具有与LC-MS法相当的灵敏度、准确性,又无需采用大型仪器,降低了试验成本.

  9. HPLC-荧光法研究阿德福韦前药的大鼠组织分布%HPLC-fluorescence study on tissue distribution of adefovir prodrugs in rats

    Institute of Scientific and Technical Information of China (English)

    黄丽华; 刘志勇; 罗婵; 母昭德

    2011-01-01

    目的:建立大鼠组织中阿德福韦(PMEA)含量测定的柱前衍生化HPLC-荧光检测法,研究新型阿德福韦前药M1、M2在Wister大鼠组织中的分布情况.方法:Benetnach C18(150 mm×4.6 mm,5μm)色谱柱,柱前衍生化;流动相A:含2%乙腈的25 mmol·L-1磷酸二氢钾缓冲溶液(5 mmol·L-1氢氧化四丁基铵,pH 6.0),流动相B:含65%乙腈的25 mmol·L-1磷酸二氢钾缓冲溶液(5 mmol·L-1氢氧化四丁基铵,pH 6.0),梯度洗脱;流速1.0 mL·min-1;柱温35℃;荧光检测波长为λex236nm和Aem420 nm.以泰诺福韦(PMPA)为内标,测定Wister大鼠肝、肾中的药物浓度.比较新型前药M1、M2与阿德福韦酯(ADV)的组织分布特点.结果:组织匀浆中PMEA线性关系、最低检测限、准确度、精密度均符合分析要求.大鼠肝组织中药物浓度:M2>M1>ADV,肾组织中药物浓度:ADV>M1>M2.结论:该方法准确、稳定,灵敏度高,可用于PMEA的检测及药动学研究.在大鼠组织中M2比M1和ADV具有更强的肝靶向性和更弱的肾靶向性,具有继续研发的价值.%Objective :To establish a pre - column derivatization HPLC - fluorescence method for the determination of adefovir ( PMEA) in rat' s tissues, and study the tissue distribution of new adefovir prodrugs in Wister rats. Methods: Pre - column derivatization;A Benetnach C18 ( 150 mm × 4. 6 mm,5 μm) column was used. Mobile phase A,2% acetonitrile in 25 mmol · L-1 potassium dihydrogen phosphate buffer with 5 mmol · L-1 TBAOH ( pH 6. 0) ;mobile phase B,65% acetonitrile in 25 mmol · L-1 potassium dihydrogen phosphate buffer with 5 mmol · L-1 TBAOH ( pH 6. 0) ,gradient elution. The flow rate was 1. 0 mL · min -1 , and the column temperature was 35 ℃ with the fuorescence detection set at λex 236 nm and λem 420 nm. Take tenofovir( PMPA) as internal standard, the concentration of PMEA in liver and kidney of Wister rats were tested, and the tissue distribution characteristic of M1 , M2 and ADV were compared. Results

  10. Method Validation for the Quantitative Analysis of Aflatoxins (B1, B2, G1, and G2) and Ochratoxin A in Processed Cereal-Based Foods by HPLC with Fluorescence Detection.

    Science.gov (United States)

    Gazioğlu, Işil; Kolak, Ufuk

    2015-01-01

    Modified AOAC 991.31 and AOAC 2000.03 methods for the simultaneous determination of total aflatoxins (AFs), aflatoxin B1, and ochratoxin A (OTA) in processed cereal-based foods by RP-HPLC coupled with fluorescence detection were validated. A KOBRA® Cell derivatization system was used to analyze total AFs. One of the modifications was the extraction procedure of mycotoxins. Both AFs and OTA were extracted with methanol-water (75+25, v/v) and purified with an immunoaffinity column before HPLC analysis. The modified methods were validated by measuring the specificity, selectivity, linearity, sensitivity, accuracy, repeatability, reproducibility, recovery, LOD, and LOQ parameters. The validated methods were successfully applied for the simultaneous determination of mycotoxins in 81 processed cereal-based foods purchased in Turkey. These rapid, sensitive, simple, and validated methods are suitable for the simultaneous determination of AFs and OTA in the processed cereal-based foods.

  11. Recognition of hypoxyloid and xylarioid Entonaema species and allied Xylaria species from a comparison of holomorphic morphology, HPLC profiles, andribosomal DNA sequences

    DEFF Research Database (Denmark)

    Stadler, M.; Fournier, J.; Læssøe, Thomas;

    2008-01-01

    liquid chromatography (HPLC) profiles with diode array and mass spectrometric detection (HPLC-DAD-MS). This methodology was used to study Entonaema pallidum. Its major stromatal constituent was identified as xylaral, a secondary metabolite known from Xylaria polymorpha. This compound was detected......, and HPLC profiles. The type specimen of E. mesentericum was located in the spirit collection of the herbarium B and found to agree morphologically with the nomenclatorily younger E. pallidum. Traces of xylaral were even detected by HPLC-DAD-MS in the spirit in which the fungus had been preserved. Entonaema...

  12. Using Technology in Social Work Practice: The mDad (Mobile Device Assisted Dad Case Study

    Directory of Open Access Journals (Sweden)

    Shawna J. Lee

    2015-07-01

    Full Text Available Mobile technology presents an exciting opportunity for social workers to reach populations that are typically underserved by interventions and services. We present one application of technology that is particularly relevant to social work practice. The mDad (Mobile Device Assisted Dad app was developed to augment existing social work practices by providing a father-friendly tool to help new fathers learn about and engage with their infants and toddlers. We discuss the process of developing the app content and conducting usability testing of the mDad app. We conclude with a discussion of the lessons learned from the mDad project, and the challenges of implementation and dissemination of technology-based interventions in community contexts.

  13. Optimization of the Analytical Method Using HPLC with Fluorescence Detection to Determine Selected Polycyclic Aromatic Compounds in Clean Water Samples; Optimizacion del Metodo Analitico mediante HPLC con Detector de Fluorescencia para la Determinacion de Ciertos Compuestos Aromaticos Policiclicos en Muestras de Aguas Limpias

    Energy Technology Data Exchange (ETDEWEB)

    Garcia Alonso, S.; Perez Pastor, R. M.

    2013-10-01

    A study on the comparison and evaluation of 3 miniaturized extraction methods for the determination of selected PACs in clear waters is presented. Three types of liquid-liquid extraction were used for chromatographic analysis by HPLC with fluorescence detection. The main objective was the optimization and development of simple, rapid and low cost methods, minimizing the use of extracting solvent volume. The work also includes a study on the scope of the methods developed at low and high levels of concentration and intermediate precision. (Author)

  14. Development and Validation of a Simultaneous RP-HPLCUV/DAD Method for Determination of Polyphenols in Gels Containing S. terebinthifolius Raddi (Anacardiaceae).

    Science.gov (United States)

    Carvalho, Melina G; Aragão, Cícero F S; Raffin, Fernanda N; de L Moura, Túlio F A

    2017-01-01

    Topical gels containing extracts of Schinus terebinthifolius have been used to treat bacterial vaginosis. It has been reported that this species has antimicrobial, anti-inflammatory and anti-ulcerogenic properties, which can be attributed to the presence of phenolic compounds. In this work, a sensitive and selective reversed-phase HPLC-UV/DAD method for the simultaneous assay of six polyphenols that could be present in S. terebinthifolius was developed. The method was shown to be accurate and precise. Peak purity and similarity index both exceeded 0.99. Calibration curves were linear over the concentration range studied, with correlation coefficients between 0.9931 and 0.9974. This method was used to determine the polyphenol content of a hydroalcoholic extract and pharmacy-compounded vaginal gel. Although the method is useful to assess the 6 phenolic compounds, some compounds could not be detected in the products. A sensitive, selective, accurate and precise reversed-phase HPLC-UV/DAD method for the simultaneous assay of six polyphenols in S. terebinthifolius Raddi Abbreviations used: RP-HPLC-UV/DAD: Reverse Phase High Performance Liquid Chromatograph with Ultraviolet and Diode Array Detector, HPLC: High Performance Liquid Chromatograph, HPLC-UV: High Performance Liquid Chromatograph with Ultraviolet Detector, ANVISA: Brazilian National Health Surveillance Agency, LOD: Limit of detection, LOQ: Limit of quantitation.

  15. Applications of HPLC-MS in compound Ilex pubescens extract study

    Institute of Scientific and Technical Information of China (English)

    Yang Yunyun; Hongwei Chen; Zhong Ming; Ma Zhiling; Teng Jiuwei; Mu Dehai

    2006-01-01

    In this paper,high performance liquid chromatography(HPLC)along with mass spectrometry (MS)and HPLC along with a diode array detector(DAD)was used to study the compound flex pubescens extract.Two ionization techniques:electro spray ionization(ESI)and atmospheric pressure chemical ionization(APCI)were used in this work.The liquid chromatograms obtained by DAD,total ion chromatograms(TIC)from positive-and negative-ion ESI-MS and the positive-and negative-ion APCI-MS were compared.The liquid chromatograms obtained by TIC from ESI-MS provided more information on chromatographic peaks than those obtained by DAD or TIC from APCI-MS.It is suggested that the fingerprints of the compound Ilex pubescens extract should be provided by the liquid chromatograms obtained by DAD together with TIC from the negative-ion ESI-MS.The molecular weights of the nine main components in an HPLC-DAD chromatogram were determined by the corresponding positive-and negative-ion ESI and the positive-and negative-ion APCI mass spectra information.In the liquid chromatogram obtained by TIC from the negative-ion ESI-MS,the molecular weights of 23 main components were determined based on the corresponding positive-and negative-ion ESI mass spectra information.

  16. Determination of telmisartan in human plasma by HPLC-fluorescence%HPLC-荧光法测定替米沙坦血药浓度的方法学考察

    Institute of Scientific and Technical Information of China (English)

    王晓红; 孙洪亮; 孔登

    2008-01-01

    目的:采用HPLC-荧光法测定替米沙坦的血浆药物浓度.方法:采用Agillent Zorbax SB-C18色谱柱(150 mm×4.6 mm,5 μm),流动相为水-乙腈(58∶42,H3PO4调pH为4~5),流速1.0 ml/min,荧光激发波长300 nm,发射波长380 nm.结果:替米沙坦浓度在5~200 ng/ml范围内与峰面积具有良好的线性关系,最低检测限为5 ng/ml.结论:该法准确可靠,适用于替米沙坦的血药浓度测定.

  17. Determination of Ivabradine in Rat Plasma by a Validated HPLC with Fluorescence Detection%HPLC-荧光检测法测定大鼠血浆伊伐布雷定浓度

    Institute of Scientific and Technical Information of China (English)

    崔学艳; 施孝金; 李中东; 钟明康

    2010-01-01

    目的 建立测定血浆中伊伐布雷定浓度的高效液相-荧光(FLU)测定方法,用于临床血浆伊伐布雷定浓度的测定.方法 血浆样品经Varian C_2柱固相萃取后采用HPLC-FLU法在Kromasil~R C_(18)柱分离,流动相采用乙腈-10 mmol·L~(-1)磷酸二氢钾(含0.3%10 mmol·L~(-1)盐酸)(28:72),流速为1 mL·min~(-1),柱温为25℃,荧光检测波长:λ_11=328 nm,λ_2=283 nm.结果 内源性杂质不干扰测定,伊伐布雷定在0.5~50μg·L~(-1)内线性良好(r=0.999 6),最低定量限为0.5 μg·L~(-1).萃取回收率为78.74%~90.77%(n=5),批内和批间精密度(RSD%)分别为2.53~6.33(n=6)和6.38~8.91(n=3).结论 建立的固相萃取-HPLC-FLU方法简便、灵敏、准确、所需血浆量少,可以用于临床血药浓度测定.

  18. Quantity Analysis of Fluorescent Brightener VBL by HPLC%荧光增白剂VBL的高效液相色谱法定量分析

    Institute of Scientific and Technical Information of China (English)

    吕双; 蒲爱军

    2015-01-01

    本文建立了高效液相色谱(HPLC)定量分析荧光增白剂VBL含量的方法.通过对色谱条件的优化,实现荧光增白剂VBL与其杂质良好分离,并采用外标法对荧光增白剂VBL的含量进行测定.

  19. 高效液相色谱荧光法测定加替沙星注射液的含量%Analysis of gatifloxacin injection by HPLC with fluorescence detection

    Institute of Scientific and Technical Information of China (English)

    黄其春; 凃文升

    2003-01-01

    目的建立高效液相色谱荧光法测定加替沙星注射液的含量方法.方法 CLC-ODS柱(15mm×6.0mm, 5μm),流动相:0.05mol/L枸橼酸-乙腈(80∶20),三乙胺调pH3.0,流速1 mL*min-1,Ex=360nm, Em=465nm,样品稀释1000倍,进样10μL.结果加替沙星在4~32ng范围内线性关系良好,回收率(99.98±0.06)%,日内、日间RSD<2%.结论该法简便、快速、准确,可用于测定加替沙星注射液的含量.%OBJECTIVE To develop an HPLC method with fluorescence detection for analysis of gatifloxacin(GTX) injection.METHOD The HPLC system consisted of CLC-ODS column(15cm×6.0mm, 5μm)and a RF-10AXL fluorescence detector. The mobile phase used composed of 0.05mol*L-1 citric acid-acetonitrile (80∶20, v/v), adjusted pH to 3.0 with triethylamine, and the flow rate was set at 1mL*min-1. The fluorescence detector was set at λex=360nm and λem=465 nm. GTX injection was diluted 1000 times with water, then 10μL was injected into HPLC System. RESULTS The method was linear at the GTX range from 4.0 ng to 32.0ng, the recovery of the method was (99.98±0.06)%(n=5), and the RSD was <2% for precision of intra-day and between-day.CONCLUSIONS The method is simple, rapid and accurate, can be used for determination of GTX.

  20. HPLC-荧光检测法测定人血浆中厄贝沙坦的浓度%Determination of Concentration of Irbesartan in Human Plasma by HPLC with Fluorescence Detection

    Institute of Scientific and Technical Information of China (English)

    王婉钢; 刘启明

    2007-01-01

    目的:建立以HPLC-荧光检测法测定人血浆中厄贝沙坦浓度的方法.方法:色谱柱为Diamonsil C18,流动相为0.02mol·L-1磷酸二氢钾-乙腈(50∶50),荧光激发波长为250nm,发射波长为375nm,流速为1.0mL·min-1,柱温为40℃.结果:厄贝沙坦检测浓度在1.0~1 000 ng·mL-1范围内线性关系良好(r=0.999 8);低、中、高3种浓度(2.0、50.0、500.00 ng·mL-1)的平均相对回收率分别为97.4%、106.1%、101.6%,日内、日间RSD均小于10%.结论:本方法简便、快速、准确、特异性和灵敏度高,可用于厄贝沙坦的血药浓度测定和药动学研究.

  1. Development and validation of a method for the determination of valproic acid in pharmaceutical formulations by high performance liquid chromatography with diode array detection (HPLC-DAD / Desenvolvimento e validação de um método para a determinação de ácido valpróico em formulações farmacêuticas por cromatografia líquida de alta eficiência acoplada com detector de conjunto de fotodiodos

    Directory of Open Access Journals (Sweden)

    Paulo Ricardo de Souza e Souza

    2013-02-01

    Full Text Available Valproic acid (VA is used as an anticonvulsant and is used in the treatment of bipolar disorder and depression. The offi cial analytical method for this drug in the compendia is gas chromatography, which is unavailable in many quality control laboratories. Here, we report a validated alternative method using high performance liquid chromatography with diode array detection (HPLC-DAD. The optimized parameters and conditions were as follows: C18 column (5 μm; 250 x 4 mm d.i.; fl ow 1.0 mL.min-1; wavelength: 210 nm; mobile phase: 55% acetonitrile (ACN in water containing 0.05% v/v trifl uoroacetic acid (TFA (v/v. The analytical parameters that were validated included the selectivity and matrix effects, linearity, repeatability and intermediate, precision, accuracy, recovery and robustness. This method identifi es VA unambiguously. In validation, the following results were obtained: good linearity in concentrations between 0.7 and 1.3 mg.mL-1 (r2 = 0.9998, relative standard deviations (RSDs of 0.68% for repeatability and 1.23% for intermediate precision, a recovery of 99.42 to 101.55% (RSD 0.14 to 0.74% and an accuracy of 100.68% (RSD = 0.79%. This method is robust to small variations in procedure. ------------------------------------------------------------------------------------- O ácido valpróico (VA é utilizado como anticonvulsivante e também na terapêutica da desordem bipolar e depressão. O método de análise deste medicamento nos compêndios ofi ciais é por cromatografi a gasosa e muitos laboratórios ofi ciais de controle de qualidade não possuem tal equipamento. A proposta deste estudo foi desenvolver e validar um método analítico por cromatografi a líquida de alta efi ciência acoplada com detector de conjunto de fotodiodos para a análise deste fármaco. Os parâmetros e condições otimizados foram: coluna C18 (5 μm; 250 x 4 mm d.i., fl uxo de 1,0 mL min-1, comprimento de onda 210 nm, fase móvel 55% ACN e 45

  2. Discrimination of Polish unifloral honeys using overall PTR-MS and HPLC fingerprints combined with chemometrics

    NARCIS (Netherlands)

    Kus, P.M.; Ruth, van S.M.

    2015-01-01

    A total of 62 honey samples of six floral origins (rapeseed, lime, heather, cornflower, buckwheat and black locust) were analysed by means of proton transfer reaction mass spectrometry (PTR-MS) and HPLC-DAD. The data were evaluated by principal component analysis and k-nearest neighbours classificat

  3. Simultaneous determination of trace levels of 10 quinolones in swine, chicken, and shrimp muscle tissues using HPLC with programmable fluorescence detection.

    Science.gov (United States)

    Zhao, Sijun; Jiang, Haiyang; Li, Xuelian; Mi, Tiejun; Li, Cun; Shen, Jianzhong

    2007-05-16

    A HPLC method using a modified sample preparation procedure was optimized and validated for the quantification of 10 quinolones (QNs), including marbofloxacin, ciprofloxacin, norfloxacin, lomefloxacin, danofloxacin, enrofloxacin, sarafloxacin, difloxacin, oxolinic acid, and flumequine, in swine, chicken, and shrimp tissues. In this method, only a small mass (residue limits (10 ng g-1) established in many countries. The method was also applied to the measurement of QN residues in commercial muscle samples. The results showed it was rapid, simple, sensitive, and suitable for use in food surveillance programs.

  4. Simultaneous determination of umbelliferone and scopoletin in Tibetan medicine Saussurea laniceps and traditional Chinese medicine Radix angelicae pubescentis using excitation-emission matrix fluorescence coupled with second-order calibration method

    Science.gov (United States)

    Wang, Li; Wu, Hai-Long; Yin, Xiao-Li; Hu, Yong; Gu, Hui-Wen; Yu, Ru-Qin

    2017-01-01

    A chemometrics-assisted excitation-emission matrix (EEM) fluorescence method is presented for simultaneous determination of umbelliferone and scopoletin in Tibetan medicine Saussurea laniceps (SL) and traditional Chinese medicine Radix angelicae pubescentis (RAP). Using the strategy of combining EEM fluorescence data with second-order calibration method based on the alternating trilinear decomposition (ATLD) algorithm, the simultaneous quantification of umbelliferone and scopoletin in the two different complex systems was achieved successfully, even in the presence of potential interferents. The pretreatment is simple due to the "second-order advantage" and the use of "mathematical separation" instead of awkward "physical or chemical separation". Satisfactory results have been achieved with the limits of detection (LODs) of umbelliferone and scopoletin being 0.06 ng mL- 1 and 0.16 ng mL- 1, respectively. The average spike recoveries of umbelliferone and scopoletin are 98.8 ± 4.3% and 102.5 ± 3.3%, respectively. Besides, HPLC-DAD method was used to further validate the presented strategy, and t-test indicates that prediction results of the two methods have no significant differences. Satisfactory experimental results imply that our method is fast, low-cost and sensitive when compared with HPLC-DAD method.

  5. Extração por fluido supercrítico de alguns inseticidas carbamatos em amostras de batata, com determinação por HPLC/fluorescência e confirmação por HPLC/espectrometria de massas

    Directory of Open Access Journals (Sweden)

    Nunes Gilvanda Silva

    2002-01-01

    Full Text Available Six supercritical fluid extraction (SFE methods were tested, by varying the following operational parameters: CO2 pressure, time and temperature of extraction, type and proportion of static modifier, and Hydromatrix®/sample rate into cell. Firstly, insecticide carbamates were extracted from spiked potatoes samples (fortification level of 0,5 mg.Kg-1 by using SPE procedures, and then final extracts were analyzed HPLC/fluorescence. Good performance was observed with SFE methods that operated with values of temperature and CO2 pressure of 50 ºC and 350 bar, respectively. Best efficiency was obtained when it was used acetonitrile as a modifier (3% on the cell volume, and Hydromatrix®/sample rate of 2:1. Static time was of 1 min; total extraction time was of 35 min; dynamic extraction was performed with 15 mL of CO2, and it was used methanol (2 mL for the dissolution of the final residue. In such conditions, pesticide recoveries varied from 72 to 94%, depending on the analyzed compound. In higher extraction temperatures, a rapid degradation was observed for some compounds, such as aldicarb and carbaryl; presence of their metabolites was further confirmed by HPLC-APCI/MS in positive mode. Detection limits for chromatographic analysis varied from 0,2 to 1,3 ng.

  6. Antimony speciation in soils: improving the detection limits using post-column pre-reduction hydride generation atomic fluorescence spectroscopy (HPLC/pre-reduction/HG-AFS).

    Science.gov (United States)

    Quiroz, Waldo; Olivares, David; Bravo, Manuel; Feldmann, Jorg; Raab, Andrea

    2011-04-15

    HG-AFS is highly sensitive and low cost detection system and its use for antimony chemical speciation coupled to HPLC is gaining popularity. However speciation analysis in soils is strongly hampered because the most efficient extractant reported in the literature (oxalic acid) strongly inhibits the generation of SbH(3) by Sb(V), the major species in this kind of matrix, severely affecting its detection limits. The purpose of this research is to reduce the detection limit of Sb(V), by using a post column on-line reduction system with l-cysteine reagent (HPLC/pre-reduction/HG-AFS). The system was optimized by experimental design, optimum conditions found were 2% (w/v) and 10°C temperature coil. Detection limits of Sb(V) and Sb(III) in oxalic acid (0.25 mol L(-1)) were improved from 0.3 and 0.1 μg L(-1) to 0.07 and 0.07 μg L(-1), respectively. The methodology developed was applied to Chilean soils, where Sb(V) was the predominant species. Copyright © 2011 Elsevier B.V. All rights reserved.

  7. 程序性细胞死亡抑制基因Dad-1在水稻和玉米中的FISH定位%Chromosomal Localization of Programmed Cell Death Suppressor OsDad-1 in Rice and Maize

    Institute of Scientific and Technical Information of China (English)

    金危危; 杨征; 宁顺斌; 李宗芸; 熊志勇; 宋运淳

    2001-01-01

    Dad-l是一种在动物和植物中都非常保守的细胞程序性死亡(PCD)抑制基因.作者利用FISH(荧光原位杂交)首次把单拷贝水稻Dad-1基因物理定位在水稻第2号染色体短臂的端部(Fig.2 A,B&C).我们还分析了它在玉米基因组中的同源序列.Southern杂交结果显示在玉米基因组中确实存在水稻Dad-1的同源序列(Fig.1).FISH进一步展示了三个杂交信号分别在玉米4、5号染色体长臂和9号染色体短臂上(Fig.2 D,E&F),其信号距着丝粒的百分距离(FL值)分别为9l、98和96.其杂交位点的位置与水稻Dad-1所处的相对位置是相似的,它们都处于染色体臂的端部.这表明在一定的程度上,Dad-1基因不仅在序列同源性上而且在所处的染色体位置上具有保守性. 水稻Dad-1基因在水稻中的杂交信号检出率(38%)高于玉米中的.这表明与玉米相比,水稻Dad-1基因的编码序列更容易与水稻染色体杂交;它与玉米中的相应序列可能只是部分同源.%The gene Dad-1 is a well-conserved programmed cell death (PCD) suppressor in animals and plants. Using fluorescence in situ hybridization (FISH), the authors physically mapped the single copy rice Dad-1 gene onto the distal region of short arm in rice chromosome 2 for the first time. The homology of it in maize genome was also analyzed. The results of Southern blot suggested that there are sequences homologous to rice Dad-1 in maize genome. FISH further showed that the three hybridization signals were detected in maize, two of them on the long arms of chromosome 4 and 5, another on the short arm of chromosome 9, respectively. And the distances from the signal to centromere (FL value) were 91%, 98% and 96%, respectively. All of the three hybridization sites of maize were similar to those of rice in their relative position, they all located at the terminal regions of the chromosome arms. It indicated that, in a certain degree, gene Dad-1 was conservative not only for

  8. Novel HPLC--Fluorescence methodology for the determination of methylglyoxal and glyoxal. Application to the analysis of monovarietal wines "Ribera del Guadiana".

    Science.gov (United States)

    Rodríguez-Cáceres, Maria Isabel; Palomino-Vasco, Mónica; Mora-Diez, Nielene; Acedo-Valenzuela, Maria Isabel

    2015-11-15

    The determination and quantification of α-dicarbonyl compounds, glyoxal and methylglyoxal, in "Ribera del Guadiana" monovarietal wines (Extremadura, Spain) without sample clean-up has been carried out by HPLC with spectrofluorimetric detection (307/371 nm). For this purpose, a derivatization step with the new reagent 3,4-diaminopyridine at pH 2 during 120 min at 90 °C has been included. Afterwards, the sample could be injected in the chromatographic system with no clean-up, during a total run time of 4 min. Several monovarietal wines (white, rosé and red) have been analyzed and the levels of these compounds for white wines were between 0.4-1.0 mg L(-1) glyoxal and 0.8-1.3 mg L(-1) methylglyoxal; and between 0.8-3.0 mg L(-1) and 0.5-1.8 mg L(-1) of glyoxal and methylglyoxal respectively, in red wines.

  9. Study of a new derivatizing reagent that improves the analysis of amino acids by HPLC with fluorescence detection: application to hydrolyzed rape bee pollen.

    Science.gov (United States)

    You, Jinmao; Liu, Lingjun; Zhao, Wenchen; Zhao, Xianen; Suo, Yourui; Wang, Honglun; Li, Yulin

    2007-04-01

    A simple and sensitive method for evaluating the chemical compositions of protein amino acids, including cystine (Cys)(2) and tryptophane (Try) has been developed, based on the use of a sensitive labeling reagent 2-(11H-benzo[alpha]-carbazol-11-yl) ethyl chloroformate (BCEC-Cl) along with fluorescence detection. The chromophore of the 1,2-benzo-3,4-dihydrocarbazole-ethyl chloroformate (BCEOC-Cl) molecule was replaced with the 2-(11H-benzo[alpha]-carbazol-11-yl) ethyl functional group, yielding the sensitive fluorescence molecule BCEC-Cl. The new reagent BCEC-Cl could then be substituted for labeling reagents commonly used in amino acid derivatization. The BCEC-amino acid derivatives exhibited very high detection sensitivities, particularly in the cases of (Cys)(2) and Try, which cannot be determined using traditional labeling reagents such as 9-fluorenyl methylchloroformate (FMOC-Cl) and ortho-phthaldialdehyde (OPA). The fluorescence detection intensities for the BCEC derivatives were compared to those obtained when using FMOC-Cl and BCEOC-Cl as labeling reagents. The ratios I (BCEC)/I (BCEOC) = 1.17-3.57, I (BCEC)/I (FMOC) = 1.13-8.21, and UV(BCEC)/UV(BCEOC) = 1.67-4.90 (where I is the fluorescence intensity and UV is the ultraviolet absorbance). Derivative separation was optimized on a Hypersil BDS C(18) column. The detection limits calculated from 1.0 pmol injections, at a signal-to-noise ratio of 3, ranged from 7.2 fmol for Try to 8.4 fmol for (Cys)(2). Excellent linear responses were observed, with coefficients of >0.9994. When coupled with high-performance liquid chromatography, the method established here allowed the development of a highly sensitive and specific method for the quantitative analysis of trace levels of amino acids including (Cys)(2) and Try from bee-collected pollen (bee pollen) samples.

  10. Determination of concentration of levodropropizine in plasma by RP-HPLC with fluorescence detector%HPLC法测定左羟丙哌嗪片血药浓度

    Institute of Scientific and Technical Information of China (English)

    毕津莲; 李湘斌

    2005-01-01

    目的建立HPLC-荧光法测定人血浆中左羟丙哌嗪浓度,用于生物利用度及药物动力学研究.方法血样经液-液萃取后,用HiQsil C18(150.0 mm×4.6mm,5μm)柱,柱温30℃,以0.1 M磷酸二氢钾(磷酸调pH值至2.51):乙腈:甲醇(82:3:15)为流动相,流速0.6mLmin,检测波长:Ex=240nm,Em=350nm进行分离.结果血浆中杂质不干扰样品测定,在3.46~886 ng/mL范围线性良好,最低可定量浓度为3.46 ng/mL(信噪比>3),萃取回收率大于80.00%,方法回收率在106.12%~109.05%,日内和日间RSD均小于10.00%.结论该法灵敏、准确,适合左羟丙哌嗪血药浓度监测.

  11. Rich Dad Poor Dad: An Entrepreneurial Approach to the Teaching of Business French

    OpenAIRE

    Sacco, Steven J.; Hammett, Joseph

    2012-01-01

    US higher education has focused on the development of new cadres of employees to the near exclusion of entrepreneurship as a career path. In this article, the authors describe an entrepreneurial approach to the teaching of Business French. The senior author served as the course instructor while the junior author was a student who completed the course. To provide an entry into the world of global entrepreneurship, the senior author selected the French translation of Robert Kiyosaki’s Rich Dad ...

  12. Development and validation of a method for the determination of low-ppb levels of macrocyclic lactones in butter, using HPLC-fluorescence.

    Science.gov (United States)

    Macedo, Fabio; Marsico, Eliane Teixeira; Conte-Júnior, Carlos Adam; de Resende, Michele Fabri; Brasil, Taila Figueiredo; Pereira Netto, Annibal Duarte

    2015-07-15

    An analytical method was developed and validated for the simultaneous determination of four macrocyclic lactones (ML) (abamectin, doramectin, ivermectin and moxidectin) in butter, using liquid chromatography with fluorescence detection. The method employed heated liquid-liquid extraction and a mixture of acetonitrile, ethyl acetate and water, with preconcentration and derivatization, to produce stable fluorescent derivatives. The chromatographic run time was validation followed international guidelines and employed fortified butter samples. The figures of merit obtained, e.g. recovery (72.4-106.5%), repeatability (8.8%), within-laboratory reproducibility (15.7%) and limits of quantification (0.09-0.16 μg kg(-1)) were satisfactory for the desired application. The application of the method to real samples showed that ML residues were present in six of the ten samples evaluated. The method proved to be simple, easy and appropriate for simultaneous determination of ML residues in butter. To our knowledge, this is the first method described for the evaluation of ML in butter.

  13. Determination of three kinds of fluorescent whitening agents in disposable cups and containers by HPLC with fluorescence detector%高效液相色谱-荧光法测定一次性纸杯和餐盒中3种荧光增白剂的含量

    Institute of Scientific and Technical Information of China (English)

    孟怀山; 李雪莉; 张盼盼; 冉文生; 姚军

    2015-01-01

    A method for the determination of fluorescent whitening agents in disposable cups and contain-ers by high performance liquid chromatography ( HPLC) with fluorescence detector was developed. The HPLC method was performed on a column of Inertsil ODS-3(250 mm × 4. 6 mm,5 μm) by gradient elu-tion using 5 mmol/L ammonium acetate and acetonitrile as the mobile phases with a flow rate of 0. 5 mL/min,and detected by the fluorescence detector at an excitation wavelength of 350 nm and an emission wavelength of 430 nm. Results showed that the three fluorescent whitening agents( B2729,220, B2730) were separated well. The recovery were 77. 4% ~94. 6%,good linearities with RSD were not more than 10%. The method is simple,accurate,sensitive and can meet the requirements of the determi-nation of fluorescent whitening agents in disposable cups and containers.%建立测定一次性纸杯和餐盒中荧光增白剂含量的高效液相色谱-荧光方法。采用Inertsil ODS-3(250 mm ×4.6 mm,5μm)为分析柱,以5 mmol/L乙酸铵溶液和乙腈为流动相,梯度洗脱,流速为0.5 mL/min,荧光激发波长为350 nm,发射波长为430 nm。结果表明,所选用的3种荧光增白剂(B2729、220、B2730)可以较好地被分离,回收率为77.4%~94.6%,相对标准偏差<10%且线性关系良好。该法简便快捷、结果准确、灵敏度高,能够满足对一次性纸杯和餐盒中荧光增白剂含量的检测。

  14. The formulation of a quantitative analysis method of HPLC with fluorescence detector for analyzing the concentration of Doxorubicin in vivo in balb/c mice%盐酸阿霉素在 balb/c 小鼠各脏器浓度的 HPLC-荧光定量分析方法的建立

    Institute of Scientific and Technical Information of China (English)

    黄佩文; 陈利鹏; 王靖; 石森林; 魏晓红

    2015-01-01

    Objective To formulate a quantitative analysis method of HPLC with fluorescence detector for analyzing the concentration of Doxorubicin in vivo in balb/c mice.Method The mobile phase has been investigated to study the possibility of daunorubicin hydrochloride as the internal standard substance with a fluorescence detector .Organic solvents were used to extract the drugs in the organ-homogenates of female balb/c mice.The recovery rate, the precision, and stability of the samples were inspected under the conditions .Findings Standard curves were successfully obtained where the R2 value was between 0.9991 and 0.9998.The recovery rate was between 90%and 110%.The precision and the stability of the samples were found desirable .Conclusion The method we formulated was found to be simple , feasible, repeatable and accurate .It can be applicable to biological samples'analysis .%目的:建立盐酸阿霉素的balb/c小鼠体内HPLC-荧光分析方法。方法采用荧光检测器,选择最佳流动相,考察盐酸柔红霉素作为内标物的可能性;使用雌性balb/c小鼠为实验动物,取出脏器匀浆,以有机溶剂提取匀浆液中的药物;并对此条件下的回收率、精密度和以及样品稳定性进行考察。结果制备的标准曲线R2值在0.9991~0.9998,回收率基本保持在90%~110%,精密度及稳定性良好。结论本文建立的分析方法简便、可行,重复性、准确性较好,适用于生物样品的体内分析。

  15. Quantitative HPLC Analysis of Rosmarinic Acid in Extracts of "Melissa officinalis" and Spectrophotometric Measurement of Their Antioxidant Activities

    Science.gov (United States)

    Canelas, Vera; da Costa, Cristina Teixeira

    2007-01-01

    The students prepare tea samples using different quantities of lemon balm leaves ("Melissa officinalis") and measure the rosmarinic acid contents by an HPLC-DAD method. The antioxidant properties of the tea samples are evaluated by a spectrophotometric method using a radical-scavenging assay with DPPH. (2,2-diphenyl-1-picrylhydrazyl). Finally the…

  16. Quantitative HPLC Analysis of Rosmarinic Acid in Extracts of "Melissa officinalis" and Spectrophotometric Measurement of Their Antioxidant Activities

    Science.gov (United States)

    Canelas, Vera; da Costa, Cristina Teixeira

    2007-01-01

    The students prepare tea samples using different quantities of lemon balm leaves ("Melissa officinalis") and measure the rosmarinic acid contents by an HPLC-DAD method. The antioxidant properties of the tea samples are evaluated by a spectrophotometric method using a radical-scavenging assay with DPPH. (2,2-diphenyl-1-picrylhydrazyl). Finally the…

  17. Kids Mean Less Shuteye for Mom, While Dad Slumbers on

    Science.gov (United States)

    ... medlineplus.gov/news/fullstory_163798.html Kids Mean Less Shuteye for Mom, While Dad Slumbers On Study ... with seven to nine hours considered optimal and less than six hours considered lacking. The men and ...

  18. Systems Integration Testing of OPUS and the new DADS

    Science.gov (United States)

    Sherbert, L. E.; Nagel, L.

    The Data Archive and Distribution System (DADS) will be entering the IDR (Ingest Distribution Redesign) era soon and more major functions will be shifting from the VMS platforms to various Unix platforms. As the first phase, Distribution, is delivered to testing, interfaces with OPUS and OTFR (On The Fly Reprocessing) will change. We will give a current overview of the OPUS/DADS/OTFR system and identify interface changes that will impact the operators and archive users.

  19. RP-HPLC-荧光法测定大鼠海马组织中5种氨基酸类神经递质%The determination of 5 amino acid neurotransmitters in hippocampus of rats by RP-HPLC with fluorescence detector

    Institute of Scientific and Technical Information of China (English)

    白钰; 张莉; 马晓丽; 孔彬; 李新霞; 李新宇

    2014-01-01

    Objective To establish a method of reverse phase high performance liquid chromatography(RP-HPLC) with fluorescence detector to determine 5 amino acid [aspartate (Asp ),glutamate (Glu ),asparagine (Aspa ), glutamine (Gln)and glycine (Gly)]neurotransmitters.Methods The amino acid neurotransmitters in hippocampus of rats were collected by methanol:water =1 ∶1 .The RP-HPLC with fluorescence detector with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC ) pre-column derivatization was used to measure the 5 amino acid neurotransmitters in hippocampus of rats.Results The 5 amino acid neurotransmitters were well and completely separated using elution within 1 5 min.There was a good linear correlation in the range of 0.5-1 00.0 μg/μL,and the correlation coefficient (r)was≥0.999 90.The detection limits of Asp,Glu,Aspa,Gln and Gly were 0.05,0.068, 0.037,0.05 and 0.02 μg/μL.The precisions [relative standard deviation(RSD)]were 0.4%-2.5%.The recoveries of added amino acids were 70.27%-1 28.20%(RSD <5%).The contents of Asp,Glu,Aspa,Gln and Gly in hippocampus of normal rats were 38.98 ±9.66,1 26.42 ±34.06,21 6.00 ±0.75,90.44 ±33.75 and (1 2.95 ± 4.42)μg/g.Conclusions RP-HPLC with fluorescence detector is quick,reliable and sensitive to measure the amino acid neurotransmitters in hippocampus of rats.%目的:采用反相高效液相色谱(RP-HPLC)-荧光检测技术建立一种用于测定脑海马组织中5种氨基酸[天门冬氨酸(Asp)、谷氨酸(Glu)、天冬酰胺(Aspa)、谷胺酰胺(Gln)、甘氨酸(Gly)]类神经递质的方法。方法采用甲醇∶水=1∶1提取大鼠脑海马组织中多种氨基酸类神经递质。用6-氨基喹啉基-N-羟基-琥珀酰亚胺氨基甲酸酯(AQC)柱前衍生RP-HPLC-荧光法测定海马组织中Asp、Glu、Aspa、Gln、Gly含量。结果5种氨基酸类神经递质在15 min内完全分离,分离效果良好;在0.5~100.0μg/μL范围有较好的线性

  20. A facile HPLC method for optical purity and quantitative measurements of phenylalanine from the hydrolyzed aspartame under different pH and temperature after its derivatization with a fluorescent reagent.

    Science.gov (United States)

    Hsien, T-J; Chen, S

    2007-07-01

    In this paper, the artificial sweetener aspartame is deliberately hydrolyzed under different pH and temperature in the matrix, and time period for the hydrolysis. The HPLC analysis is then performed to quantitatively measure the amount and the optical purity of phenylalanine produced as a result of hydrolysis in the matrix after its functionalization with a fluorescent reagent. The results show that the amount of phenylalanine in the matrix is affected by the pH variation during the hydrolysis and found increased in low pH conditions. High temperature or long time periods for the decomposition also increases the amount, which indicates that beverages and foods containing aspartame as a sweetener may not be safe for phenylketonuria patients to consume if they are stored under these conditions. Conversely, the optical purity of phenylalanine, expressed as the percentage of D: -enantiomer, is not affected by pH variations. However, it decreases as the length of time elapsed is increased or surrounding temperature is elevated during the decomposition.

  1. A rapid method for the determination of dopamine in porcine muscle by pre-column derivatization and HPLC with fluorescence detection

    Institute of Scientific and Technical Information of China (English)

    Hong-Xia Zhao; Hui Mu; Yan-Hong Bai; Hu Yu; Ying-Mei Hu

    2011-01-01

    A rapid method has been developed based on the sample preparation procedure named as QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe), combined with reversed-phase high performance liquid chromatography with fluorescence detector and C18 column after precolumn derivatization using o-phthalaldehyde and 2-mercaptoethanol to determine dopamine in porcine muscle. Methanol and deionized water (0.1% acetic acid, v/v) with a ratio of 60:40 was used as mobile phase. The flow rate was 0.8 mL/min and dopamine was eluted within 15 min. The linearity range was 0.003-8 μg/mL with r=0.9992. The detection limit for dopamine was 4 μg/kg and the quantification limit was 9 μg/kg. Recovery studies were carried out at 0.1, 0.5 and 1.0 mg/kg fortification levels and the average recoveries obtained ranged from 90.4% to 98.2% with relative standard deviations between 3.5% and 8.1%. The method was found to be suitable for detection of dopamine in animal product tissues at the maximum residue level.

  2. Sulfonate-tagged 1,4-diazabutadiene (DAD(S)) ligands and their noble-metal complexes--synthesis, characterization and immobilization in ionic liquids.

    Science.gov (United States)

    Oelkers, Benjamin; Sundermeyer, Jörg

    2011-12-21

    A series of sulfonate-tagged 1,4-diazabutadiene (DAD(S)) ligands was prepared as salts with typical ionic liquid (IL) cations ([EMIM](+), [BMIM](+), [BMMIM](+), Bu(4)N(+), Bu(3)PMe(+), [Gua-4,4-4,4-4,1](+)). Complexation behaviour of the ligands was investigated by preparing complexes of the types [BMMIM](2)[MCl(2)(DAD(S))] (M = Pd, Pt), [BMMIM][Rh(COD)(DAD(S))] and [BMMIM](2)[Mo(CO)(4)(DAD(S))]. Using UV-Vis spectroscopy, the latter sulfonate-tagged chromophore was shown to be well soluble in the sulfonate IL [BMIM]OTf and completely insoluble in toluene, resulting in perfect immobilization. The crystal structures of [HNEt(3)](2)[2,6-Me(2)-Me-DAD(S)], [BMIM](2)[2,6-Me(2)-Me-DAD(S)], [BMMIM](2)[2,4,6-Me(3)-Me-DAD(S)], [BMMIM](2)[2,6-iPr(2)-Me-DAD(S)] and [HNEt(3)](2)[PdCl(2)(2,6-Me(2)-Me-DAD(S))] were determined. Regarding the diimine fragment, they show geometries similar to the respective non-sulfonated parent compounds.

  3. HPLC-fluorescence determination of vitamin B2 content in human blood plasma with solid phase extraction%固相萃取结合HPLC-荧光法测定人血浆中维生素B2含量

    Institute of Scientific and Technical Information of China (English)

    田兰; 孔彬; 杜晨松; 居来提·安尼瓦尔; 买买提艾力·乌马尔; 巴吐尔·买买提明

    2012-01-01

    目的:建立固相萃取结合HPLC-荧光法测定人血浆中维生素B2含量的方法.方法:采用SiroccoTM96孔蛋白沉淀板和正压固相萃取装置处理样品,采用SymmetryshieldTMRP-C18(250 mm×4.6 mm,5μm)色谱柱,流动相为甲醇-5 mmol·L-1乙酸铵(35:65),流速为0.6 mL·min-1,荧光检测器检测(激发波长:450 nm,发射波长:520 nm).结果:维生素B2线性范围为0.2~10ng·mL-1(r=0.9937).定量下限为0.17 ng·mL1,日内精密度(RSD)为2.0%~4.8%,日间精密度(RSD)为3.8%,方法回收率为101.5%~116.0%(n=3),提取回收率为91.8% ~ 115.1%.结论:所建方法采用96孔蛋白沉淀装置自动处理样品,高效液相色谱-荧光法测定,可用于临床血浆样品中维生素B2含量的测定.%Objectives: To establish a method for the determination of the content of vitamin B2 in human blood plasma by HPLC -fluorescence detection combined with solid phase extraction. Method:Sirocco?96 precipitation plate and the positive pressure - 96 processor were used for sample pre - treatment. The chromatographic column was Symmetryshield? RP - C18(250 mm ×4. 6 mm,5 μm) ,the mobile phase was 35% methanol and 65% 5 mmol · L -1 ammonium acetate solution at a flow rate of 0. 6 mL · min -1 . The spectro - photofluorimeter was set at wavelength of 450 nm for excitation and 520 nm for emission. Results:The linear ranges were 0. 2 - 10 ng · mL -1 ( r = 0. 9937). The lowest limit of quantification was 0. 17 ng · mL-1. The intra - day RSD were 2. 0% -4. 8% ,the inter - day RSD was 3.8%. The relative recoveries were between 101. 5% and 116. 0% (n = 3 ) . The recovery of extraction were 91. 8% -115. 1%. Conclusions: A 96 - well plate automation - compatible precipitation plate was used to dispose the samples, which determined the content of vitamin B2 by the HPLC - fluorescence detector. This method can be used for determining of vitamin B2 in clinical plasma samples.

  4. Extraction, Separation, and Identification of Phenolic Compounds in Virgin Olive Oil by HPLC-DAD and HPLC-MS

    OpenAIRE

    Maria Tasioula-Margari; Eleftheria Tsabolatidou

    2015-01-01

    The aim of this study was to evaluate the recovery of individual phenolic compounds extracted from virgin olive oil (VOO), from different Greek olive varieties. Sufficient recoveries (90%) of all individual phenolic compounds were obtained using methanol as an extraction solvent, acetonitrile for residue solubilization, and two washing steps with hexane. Moreover, in order to elucidate structural characteristics of phenolic compounds in VOO, high performance liquid chromatography with a diode...

  5. Determination of MDMA and MDA in rat urine by semi-micro column HPLC-fluorescence detection with DBD-F and their monitoring after MDMA administration to rat.

    Science.gov (United States)

    Wada, Mitsuhiro; Nakamura, Shinichi; Tomita, Mamoru; Nakashima, Mihoko N; Nakashima, Kenichiro

    2005-01-01

    A simultaneous semi-micro column HPLC method with fluorescence detection of abused drugs, such as 3,4-methylenedioxymethamphetamine (MDMA), 3,4-methylenedioxyamphetamine (MDA), amphetamine (AP) and methamphetamine (MP) in rat urine was examined by using 4-(N,N-dimethylaminosulphonyl)-7-fluoro-1,2,3-benzoxadiazole (DBD-F) as a labelling reagent and alpha-phenylethylamine as an internal standard (IS). A sample (50 microL) of rat urine was added to 5 microL IS and 100 microL 100 mmol/L borate buffer (pH 12) and extracted with 1.5 mL n-hexane. After evaporation, 50 microL 75 mmol/L borate buffer (pH 8.5) and 50 microL 20 mmol/L DBD-F in CH3CN were added to the residue and mixed well. The resultant solution was heated for 20 min at 80 degrees C and then cooled in an ice bath. A good separation of DBD-derivatives could be achieved within 45 min using a semi-micro ODS column with an eluent of CH3CN/CH3OH/10 mmol/L imidazole-HNO3 buffer (pH 7.0) (= 45:5:50, v/v/v %). The DBD derivatives were monitored at 565 nm with an excitation at 470 nm. The calibration curves showed good linearity (r = 0.997) with 0.5-15 ng/mL detection limits at a S/N ratio of 3. MDMA and MDA in rat urine could be monitored for 15 h after a single administration of MDMA to rat (2.0 mg/kg, i.p.). The concentrations for MDMA and MDA (n = 3) were 0.13-160.1 and 0.17-10.9 microg/mL, respectively.

  6. 氯雷他定血药浓度的HPLC荧光检测法及生物等效性研究%Determination of loratadine in human plasma by HPLC with fluorescence detector and study on its bioavailability

    Institute of Scientific and Technical Information of China (English)

    徐晓杰; 尚尔鑫; 裘福荣; 冒国光; 相秉仁

    2004-01-01

    目的建立高效液相色谱法荧光检测血浆中氯雷他定(loratadine)含量的方法,以评价氯雷他定的相对生物利用度.方法色谱柱为Alltech C18,4.6 mm×150 mm;流动相为乙腈-水-冰醋酸-三乙胺(90∶100∶6∶0.15);流速为1 mL·min-1;荧光检测器测定波长,Ex=274 nm,Em=450 nm.结果 HPLC测定线性范围为0.2~30 μg·L-1,最低定量限0.2 μg·L-1,方法回收率为96%~98%.人体生物利用度结果表明,实验片、胶囊与对照片间的AUC,Tmax,Cmax和T1/2β均无显著性差异(P>0.05),两者的相对生物利用度分别为107%±17%和100%±14%.AUC和Cmax经可信区间法检验生物等效.结论 3种制剂生物等效.%Aim To establish an HPLC-fluorescence method for determination of loratadine in human plasma and evaluate its relative bioavailability. Methods An Alltech-C18 column and a mobile phase of acetonitrile-water-glacial acetic acid-triethylamine (90∶100∶6∶0.15) were used. The fluorescence detector was set at Ex 274 nm, Em 450 nm. The flow rate was 1 mL·min-1. Results The calibration curve was linear over a concentration range of 0.2-30 μg·L-1. The limit of quantification was 0.2 μg·L-1. The average method recoveries varied from 96% to 98%. The results showed AUC, Tmax, Cmax and T1/2β between the testing tablets, testing capsules and reference tablets had no significant difference (P>0.05). Relative bioavailabilities were 107%±17% and 100%±14% respectively. Conclusion The three formulations were bioequivalent.

  7. On-line study of flavonoids of Trollius chinensis Bunge binding to DNA with ethidium bromide using a novel combination of chromatographic, mass spectrometric and fluorescence techniques.

    Science.gov (United States)

    Song, Zhiling; Wang, Hong; Ren, Biao; Zhang, Baobao; Hashi, Yuki; Chen, Shizhong

    2013-03-22

    The study of the interaction between drugs and DNA is an important way to understand the role of drug molecules. A novel online analytical method for this purpose combining high-performance liquid chromatography-diode array detector-electrospray ionization-ion-trap time-of-flight mass spectrometry (HPLC-DAD-ESI-IT-TOF-MS(n)) and DNA-ethidium bromide detection with a fluorescence detector (DNA-EB-FLD) was firstly developed, which could rapidly identify the chemical constituents and obtain the profile related to DNA binding activity. This method has been applied for a precise or probable identification of the chemical constituents by ultraviolet (UV) absorption and MS(n) data analysis, while the DNA binding profile has been characterized by directly measuring the fluorescence intensity of compound-DNA-EB. Using this method, Trollius chinensis Bunge was studied and 18 constituents were identified by MS(n) data; six of them (4'-methoxy-2″-O-(2‴-methylbutyryl)vitexin,2″-O-(3‴-methoxycaffeoyl)vitexin) and 4'-methoxy-2″-O-(2‴-methylbutyryl)orientin,acacetin-7-O-rutinoside,quercetin-3-O-xylosylglucoside,quercetin-3-O-arabinosylglucoside) were identified for the first time in T. chinensis Bunge, and 16 constituents accounted for its activity of binding to DNA. The established (HPLC-DAD-ESI-IT-TOF-MS(n) DNA-EB-FLD) system has proved to offer a useful strategy for correlating the chemical profile with the binding to DNA activities of the components without their isolation and purification, and may be used for multicomponent analysis of active substances in other herbs.

  8. HPLC/DAD determination of rosmarinic acid in Salvia officinalis: sample preparation optimization by factorial design

    Energy Technology Data Exchange (ETDEWEB)

    Oliveira, Karina B. de [Universidade Federal do Parana (UFPR), Curitiba, PR (Brazil). Dept. de Farmacia; Oliveira, Bras H. de, E-mail: bho@ufpr.br [Universidade Federal do Parana (UFPR), Curitiba, PR (Brazil). Dept. de Quimica

    2013-01-15

    Sage (Salvia officinalis) contains high amounts of the biologically active rosmarinic acid (RA) and other polyphenolic compounds. RA is easily oxidized, and may undergo degradation during sample preparation for analysis. The objective of this work was to develop and validate an analytical procedure for determination of RA in sage, using factorial design of experiments for optimizing sample preparation. The statistically significant variables for improving RA extraction yield were determined initially and then used in the optimization step, using central composite design (CCD). The analytical method was then fully validated, and used for the analysis of commercial samples of sage. The optimized procedure involved extraction with aqueous methanol (40%) containing an antioxidant mixture (ascorbic acid and ethylenediaminetetraacetic acid (EDTA)), with sonication at 45 deg C for 20 min. The samples were then injected in a system containing a C{sub 18} column, using methanol (A) and 0.1% phosphoric acid in water (B) in step gradient mode (45A:55B, 0-5 min; 80A:20B, 5-10 min) with flow rate of 1.0 mL min-1 and detection at 330 nm. Using this conditions, RA concentrations were 50% higher when compared to extractions without antioxidants (98.94 {+-} 1.07% recovery). Auto-oxidation of RA during sample extraction was prevented by the use of antioxidants resulting in more reliable analytical results. The method was then used for the analysis of commercial samples of sage. (author)

  9. Simultaneous Determination of Seven Alkaloids in Fufang Zhenzhu Tiaozhi Capsule by HPLC Coupled with DAD

    Institute of Scientific and Technical Information of China (English)

    CHEN Yuan-yuan; GUO Jiao; FAN Hui; HUANG Li-hua

    2012-01-01

    Objective To establish a reverse-phase liquid chromatography method for the determination of seven alkaloids (magnoflorine,columbamine,jatrorrhizine,epiberberine,coptisine,palmatine,and berberine) in Fufang Zhenzhu Tiaozhi Capsule.Methods Chromatography was performed on a Dionex Acclaim C18 column (250 mm × 4.6 mm,5.0 μm) at 30 ℃.The mobile phase was composed of acetonitrile-potassium dihydrogen phosphate solution (0.015 mol/L,40:60,including 1.7 g/L sodium dodecyl sulfate and phosphoric acid used to regulate pH value to 3.0),with a flow rate of 1.0 mL/min.The detection wavelength was 270 nm.Results The calibration curves of magnoflorine,columbamine,jatrorrhizine,epiberberine,coptisine,palmatine,and berberine were linear in the range of 1.07-10.65,0.78-7.55,0.75-7.50,1.60-15.95,2.69-26.85,2.31-23.10,and 6.04-60.40 mg/mL.The average recoveries of magnoflorine,columbamine,jatrorrhizine,epiberberine,coptisine,palmatine,and berberine were 101.0%,101.2%,100.1%,100.0%,100.1%,101.1%,and 99.7%,respectively.Conclusion The method could be used for the quantitative determination of the preparation.

  10. Comprehensive chemical analysis of Schisandra chinensis by HPLC-DAD-MS combined with chemometrics.

    Science.gov (United States)

    Liu, Haitao; Lai, Hongwu; Jia, Xinyue; Liu, Jiushi; Zhang, Zhao; Qi, Yaodong; Zhang, Jin; Song, Junbin; Wu, Chongming; Zhang, Bengang; Xiao, Peigen

    2013-09-15

    The fruit of Schisandra chinensis, namely "Wuweizi" in China, is a well-known herbal medicine and health food. In this paper, an accurate and reliable high performance liquid chromatography coupled with diode array detection and mass spectrometry was developed for quality evaluation of Wuweizi. Nine lignans, including schisandrol A, schisandrol B, angeloylgomisin H, gomisin G, schisantherin A, schisanhenol, schisandrin A, schisandrin B, and schisandrin C were determined simultaneously in forty-three batches of Wuweizi samples collected from different localities. Thirty-six common peaks were unequivocally identified or tentatively assigned by comparing their mass spectrometric data with reference compounds, self-established compound library and published literatures. And the thirty-six common peaks were selected as characteristic peaks to assess the similarity of chromatographic fingerprinting of these Wuweizi samples. Moreover, hierarchical clustering analysis and principal components analysis were successfully applied to demonstrate the variability of these Wuweizi samples. The results indicated the content of nine investigated lignans varied greatly among the samples, and samples collected from different localities could be discriminated. Furthermore, schisandrol A, schisandrol B, schisandrin B, and schisandrin C were found to chemical marker for evaluating the quality of Wuweizi.

  11. Expression of defender against apoptotic death (DAD-1) in iris and dianthus petals

    NARCIS (Netherlands)

    Kop, van der D.A.M.; Ruys, G.; Dees, D.; Schoot, van der C.; Boer, de A.D.; Doorn, van W.G.

    2003-01-01

    The gene defender against apoptotic death (DAD-1) prevents programmed cell death in animal cells. We investigated the expression pattern of DAD-1 in petals of iris (Iris x hollandica cv. Blue Magic) and carnation (Dianthus caryophyllus cv. Etarro). DAD-1 expression in Iris petals was strongly reduce

  12. Simultaneous analysis of six aristolochic acids and five aristolactams in herbal plants and their preparations by high-performance liquid chromatography-diode array detection-fluorescence detection.

    Science.gov (United States)

    Yuan, Jinbin; Liu, Qian; Zhu, Weifeng; Ding, Li; Tang, Fei; Yao, Shouzhuo

    2008-02-22

    Aristolochic acid analogues, including aristolochic acids (AAs) and aristolactams (ALs), are known to be nephrotoxic, carcinogenic and mutagenic. In this paper, a high-performance liquid chromatography-diode array detection-fluorescence detection (HPLC-DAD-FLD) method was developed for the simultaneous determination of six AAs together with five ALs. Baseline separation was obtained on an ODS C18 analytical column with 0.2% HAc/methanol gradient elution. The hyphenation of DAD and FLD allows the method to directly meet the analysis requirements of most herbal plants with high sensitivity and selectivity. For trace analysis, aristolochic acids were reduced to their corresponding aritstolactams in acidic solution containing iron powder, and then high sensitive detection and quantification were carried out. The method was successfully validated in the matrices of various Aristolochiaceae plants and their preparations. Linearities of around 3-4 orders of magnitude were obtained with correlation coefficients exceeding 0.9970. The detection limits were decreased to 0.2ng/ml. Satisfactory intra-day and inter-day precisions were achieved with RSDs less than 5.74%, and the average recovery factors were in the range of 94.5-99.2%.

  13. A Method of Hepatocyte Extraction Conjugated with HPLC is Established for Screening Potential Active Components in Chinese Medicines—Probing Herba Artemisiae Scopariae as an Exemplifying Approach

    Directory of Open Access Journals (Sweden)

    Hong-Wei Fan

    2012-02-01

    Full Text Available In order to establish an effective and quick method for screening potential bioactive compounds in Traditional Chinese Medicines (TCMs, hepatocytes were employed for extracting either bifendate, a clinical medicine for liver diseases, or chemicals in Herba Artemisiae Scopariae (A. Scopariae, a commonly used traditional Chinese medicine for remedying liver diseases such as hepatitis induced by viruses, chemicals or alcohol. After hepatocyte extraction the compounds were analyzed by HPLC, therefore this method was referrred to as hepatocyte extraction conjugated with HPLC (HE-HPLC. In the first part of this study, HE-HPLC showed that bifendate was extracted by hepatocytes and detected by HPLC-DAD which indicated the feasibility of this method. Then in the second part of the study, the potential active components in the A. scopariae extract were studied using HE-HPLC. Six chemicals in the A. scopariae extract, which could bind to hepatocytes in vitro, were detected by HPLC-DAD and three were identified as 7-hydroxy-coumarin (7-OH-C, capillartemisin A and 7-methoxy-coumarin, respectively. In vitro assays showed that 7-OH-C protected HL-7702 hepatocytes from H2O2 injury. The results indicated that these compounds could be extracted by hepatocytes, could be detected by HPLC and more importantly were bioactive. It is suggested that HE-HPLC is a useful method for screening potent active components in Chinese medicines used to treat liver diseases.

  14. Mom and Dad: An Untapped Source for Career Information.

    Science.gov (United States)

    Shapiro, Ronald G.

    Obtaining and evaluating career information is a beneficial activity for high school students. Although a potential source of this information is parents, students do not necessarily have meaningful career discussions with their parents. An activity, Thats My Mom / Thats My Dad was developed to encourage more parent-student career decisions. In…

  15. LC-DAD-ESI/MS(n) determination of direct condensation flavanol-anthocyanin adducts in pressure extracted pomegranate (Punica granatum L.) juice.

    Science.gov (United States)

    Sentandreu, Enrique; Navarro, Jose L; Sendra, Jose M

    2010-10-13

    Pomegranate (Punica granatum L.) juice, obtained by pressure extraction of the whole fruit, has been analyzed for its flavanol-anthocyanin adduct content using reversed-phase liquid chromatography with diode array detection, coupled to mass spectrometry (ion trap) with electrospray ionization (HPLC-DAD-ESI/MS(n)), operating in positive ion mode. A total of 35 dimers have been detected, consisting of mono- and disubstituted hexoside derivatives of the adducts between the flavan-3-ols (epi)gallocatechin, (epi)catechin and (epi)afzelechin and the anthocyanidins delphinidin, cyanidin and pelargonidin. In addition, evidence is given for the presence of additional anthocyanin-flavanol adducts in this juice.

  16. DADS Suppresses Human Esophageal Xenograft Tumors through RAF/MEK/ERK and Mitochondria-Dependent Pathways

    Directory of Open Access Journals (Sweden)

    Xiaoran Yin

    2014-07-01

    Full Text Available Diallyl disulfide (DADS is a natural organosulfur compound isolated from garlic. DADS has various biological properties, including anticancer, antiangiogenic, and antioxidant effects. However, the anticancer mechanisms of DADS in human esophageal carcinoma have not been elucidated, especially in vivo. In this study, MTT assay showed that DADS significantly reduced cell viability in human esophageal carcinoma ECA109 cells, but was relatively less toxic in normal liver cells. The pro–apoptotic effect of DADS on ECA109 cells was detected by Annexin V-FITC/propidium iodide (PI staining. Flow cytometry analysis showed that DADS promoted apoptosis in a dose-dependent manner and the apoptosis rate could be decreased by caspase-3 inhibitor Ac-DEVD-CHO. Xenograft study in nude mice showed that DADS treatment inhibited the growth of ECA109 tumor in both 20 and 40 mg/kg DADS groups without obvious side effects. DADS inhibited ECA109 tumor proliferation by down-regulating proliferation cell nuclear antigen (PCNA. DADS induced apoptosis by activating a mitochondria-dependent pathway with the executor of caspase-3, increasing p53 level and Bax/Bcl-2 ratio, and downregulating the RAF/MEK/ERK pathway in ECA109 xenograft tumosr. Based on studies in cell culture and animal models, the findings here indicate that DADS is an effective and safe anti-cancer agent for esophageal carcinoma.

  17. HPLC–DAD of phenolics in bryophytes Lunularia cruciata, Brachytheciastrum velutinum and Kindbergia praelonga

    Directory of Open Access Journals (Sweden)

    NEBOJSA JOCKOVIC

    2008-12-01

    Full Text Available The chemistry of bryophytes is not well known. The available data indicate interesting chemical constitutions of some bryophyte species, i.e., active and new compounds are to be found within bryophytes, especially liverworts. In this study, one liverwort and two moss species were studied: Lunularia cruciata (L. Dumort, Brachytheciastrum velutinum (Hedw Ignatov & Huttunen and Kindbergia praelonga (Hedw Ochyra. The phenolic compositions of these bryophyte species have not hitherto been reported. Their methanolic extracts were analyzed by reversed-phase HPLC, coupled to a diode-array detector (DAD. Luteolin-7-O-glucoside and quercetin were found in the L. cruciata extract. The extract obtained from B. velutinum contained four phenolic acids (4-O-caffeoylquinic, 5-O-caffeoylquinic, caffeic and ellagic acids and three flavonoids (apigenin-7-O-glucoside, luteolin and apigenin. The K. praelonga extract was characterized by the presence of several phenolic acids and their derivatives (4-O-caffeoylquinic, 5-O-caffeoylquinic, caffeic, p-coumaric, ferulic and ellagic acids, and caffeic and p-coumaric acid derivatives and three flavonoids (apigenin-7-O-glucoside, luteolin, apigenin and an un-identified flavanone.

  18. Entrevista com a professora Dadá Martins

    Directory of Open Access Journals (Sweden)

    Demian Garcia Castro

    2015-01-01

    Full Text Available A professora Maria Adailza Martins de Albuquerque, conhecida como Dadá Martins, presenteou a Giramundo com uma conversa muito estimulante. Natural do Ceará, com experiência de atuação profissional na rede pública do estado de São Paulo, Dadá Martins é professora e pesquisadora da Universidade Federal da Paraíba - UFPB e atualmente realiza pós-doutorado na Universidade de Lisboa - UL, em Portugal. Vem dedicando-se, desde seu doutorado, a pesquisas em história da Geografia Escolar no Brasil, com especial atenção aos livros didáticos do século XIX. A entrevista aqui apresentada se realizou em conexão direta Rio-Lisboa, via plataforma digital, que não prejudicou em nada a expressividade, o conhecimento e o envolvimento de Dadá com todos os temas propostos. Fica o agradecimento à professora, por seu pronto aceite em dialogar e contribuir com nossa revista, e a satisfação pelo resultado tão interessante, que se pode conferir a seguir.

  19. Qualitative and quantitative analysis of the major constituents in traditional Chinese medicine Danmu injection using LC-ESI-MS(n) and LC-DAD.

    Science.gov (United States)

    Zhu, Fen-Xia; Wang, Jing-Jing; Li, Xiu-Feng; Sun, E; Jia, Xiao-Bin

    2014-07-01

    Danmu injection, a traditional Chinese medicine (TCM) preparation made from Nauclea officinalis, has been commonly used for the treatment of cold, fever, swelling of throat in China. However, the chemical constituents in Danmu injection have not been clarified yet. a HPLC/DAD/ESI-MS(n) method was developed for qualitative and quantitative analysis of the components in Danmu injection. The chromatographic separation was performed on a Welch Material XB-C18 (4.6mm × 250mm, 5μm) using gradient elution with acetonitrile (A) and water containing 0.1% formic acid (B) as mobile phase at a flow rate of 1.0 ml/min. Twenty-five compounds, including phenolic acid and phenol glycoside, iridous glycoside and glycoalkaloid were identified or tentatively deduced on the base of their retention behaviors, UV absorption, MS and MS(n) data with those elucidated references or literature. In addition, eleven compounds were simultaneously determined by HPLC-DAD, which was validated and successfully applied for determination of major components in Danmu injection. The results suggested that the established qualitative and quantitative method would be a powerful and reliable analytical tool for the characterization of multi-constituent in complex chemical system and quality control of Danmu injection.

  20. Very small injected samples to study chloroquine and quinine in human serum using capillary-LC and native fluorescence

    OpenAIRE

    Ibrahim, H.; Bouajila, J.; Siri, N.; Rozing, G.; Nepveu, Françoise; Couderc, F.

    2007-01-01

    A comparison between HPLC with conventional fluorescence detection and capillary-LC (mu HPLC) with native laser-induced fluorescence (LIF) detection was done to determine chloroquine (CQ) and quinine (Q) in human serum. HPLC experiments were run with parameters of the conventional fluorimeter set at the highest level of sensitivity. Results were compared with those obtained on mu HPLC coupled to a ZETALIF (He-Cd 325 nm) detector which provided a 50-fold increase in sensitivity. In mu HPLC-LIF...

  1. 用于HPLC法测定多胺类化合物的4种荧光衍生试剂的合成及其性质%Synthesis and Properties of Four Fluorescent Reagents for HPLC Determination of Polyamines

    Institute of Scientific and Technical Information of China (English)

    许泳吉; 王永智

    2011-01-01

    设计并合成了4种相对分子质量较小、疏水性较低的荧光衍生试剂:邻氨基苯甲酸酐、N-甲基邻氨基苯甲酸酐、2-氨基烟酸活性酯和水杨酸活性酯.系统研究了这些衍生试剂与己胺的反应性能,并合成了相应的衍生物标准品,测定了衍生物的荧光性质.结果表明:4种衍生试剂中邻氨基苯甲酸酐适合用作高效液相色谱荧光检测多胺的衍生试剂.%Four fluorescent reagents, anthranilic acid anhydride, N-methyl-σ-aminobenzoic acid anhydride, 2-aminonicotinic acid active ester and salicylic acid active ester, were designed and synthesized. The reaction properties between these reagents and hexylamine were studied. The corresponding standard derivatives were synthesized and the fluorescent properties of these derivatives were also measured. The results showed that anthranilic acid anhydride was a suitable fluorescent reagent for detection of polyamine by HPLC.

  2. Analysis of natural red dyes (cochineal) in textiles of historical importance using HPLC and multivariate data analysis.

    Science.gov (United States)

    Serrano, Ana; Sousa, Micaela M; Hallett, Jessica; Lopes, João A; Oliveira, M Conceição

    2011-08-01

    A new analytical approach based on high-performance liquid chromatography with diode array detector (HPLC-DAD) and multivariate data analysis was applied and assessed for analyzing the red dye extracted from cochineal insects, used in precious historical textiles. The most widely used method of analysis involves quantification of specific minor compounds (markers), using HPLC-DAD. However, variation in the cochineal markers concentration, use of aggressive dye extraction methods and poor resolution of HPLC chromatograms can compromise the identification of the precise insect species used in the textiles. In this study, a soft extraction method combined with a new dye recovery treatment was developed, capable of yielding HPLC chromatograms with good resolution, for the first time, for historical cochineal-dyed textiles. After principal components analysis (PCA) and mass spectrometry (MS), it was possible to identify the cochineal species used in these textiles, in contrast to the accepted method of analysis. In order to compare both methodologies, 7 cochineal species and 63 historical cochineal insect specimens were analyzed using the two approaches, and then compared with the results for 15 historical textiles in order to assess their applicability to real complex samples. The methodology developed here was shown to provide more accurate and consistent information than the traditional method. Almost all of the historical textiles were dyed with Porphyrophora sp. insects. These results emphasize the importance of adopting the proposed methodology for future research on cochineal (and related red dyes). Mild extraction methods and HPLC-DAD/MS(n) analysis yield distinctive profiles, which, in combination with a PCA reference database, are a powerful tool for identifying red insect dyes.

  3. Annotating abstract pronominal anaphora in the DAD project

    DEFF Research Database (Denmark)

    Navarretta, Costanza; Olsen, Sussi Anni

    2008-01-01

    n this paper we present an extension of the MATE/GNOME annotation scheme for anaphora (Poesio 2004) which accounts for abstract anaphora in Danish and Italian. By abstract anaphora it is here meant pronouns whose linguistic antecedents are verbal phrases, clauses and discourse segments....... The extended scheme, which we call the DAD annotation scheme, allows to annotate information about abstract anaphora which is important to investigate their use, see Webber (1988), Gundel et al. (2003), Navarretta (2004) and which can influence their automatic treatment. Intercoder agreement scores obtained...

  4. Determination of Tetracycline Antibiotics Residue by HPLC with Precolumn Derivatization Fluorescence Enhancement Effect%四环素类抗生素的柱前衍生化-HPLC分离-荧光增敏检测法

    Institute of Scientific and Technical Information of China (English)

    郑洁

    2012-01-01

    本文通过柱前衍生的方法,并通过对四种常用的四环素类抗生素残留检测方法的比较研究,建立了一种高效灵敏的四环素类抗生素残留的高效液相色谱检测的方法,适合于日常检测需要的HPLC - FLD分离检测分析方法.

  5. Determination of alcohols by HPLC with 1,4-dimethylcarbazole-9-y1-propionic acid as fluorescence derivative reagent%以1,4-二甲基咔唑-9-丙酸为荧光衍生化试剂高效液相色谱法测定醇

    Institute of Scientific and Technical Information of China (English)

    劳文剑; 宋翠华; 尤进茂; 欧庆瑜

    2013-01-01

    1,4-二甲基咔唑-9-丙酸与间苯二磺酰氯反应生成混合酸酐,该混合酸酐能够衍生化18个碳以下的直链脂肪伯醇、6个碳以下的仲醇,以及叔丁醇和胆固醇;将荧光衍生物用反相C18液相色谱柱进行分离,在室温下10 min完成衍生化反应,衍生物24 h内基本稳定.基于此,建立了血清中醇的高效液相色谱测定方法.%Pre-column derivative reagent 1,4-dimethyl-carbazole-9-yl-propionic acid was allowed to couple with benzenedisulfonyl dichloride affording the mixed acidic anhydride. Resultant mixed acidic anhydride was used to derivate straight-chain aliphatic primary alcohols with less than 18 carbons and secondary alcohols with less than 6 carbons, as well as tert-butanol and cholesterol. As-obtained fluorescence derivatives were separated on reversed-phase C18 high-performance liquid chromatography (HPLC) column, with which the derivatization was finished in 10 min at room temperature and relevant derivatives were stable within 24 h. This makes it feasible to establish a HPLC method to determine alcohols in human serum with 1,4-dimethyl-carbazole-9-yl-propionic acid as the fluorescence derivative reagent.

  6. The History and Future of STScI DADS

    Science.gov (United States)

    Gaffney, N.; Kidwell, R.; Kyprianou, M.; Abney, F.

    2010-12-01

    For more than 15 years, the archive at STScI has revolved around DADS (Data Archive and Distribution System). Originally a software product delivered by a third party vendor to STScI, this system has evolved to keep up with new technologies and users needs. This evolution has encompassed changes in the core language from FORTRAN to Java, changes of OSes including moving from VMS to *nix, several transitions of the archive storage media, and the inclusion of on the fly processing for current instruments as well as reprocessing of heritage instruments. This evolution has also been driven by other missions archived by DADS (FUSE and Kepler) and expansion into the future with our JWST development. Currently we are transitioning from Sybase to SQLServer as our DB platform and Solaris to RHEL as our base OS. From this history and near future plans, we will present our list of both lessons learned and share common ideas needed to make more flexible and perhaps shareable archive components for the astronomical community.

  7. RP-HPLC Determination of 1,3-Dideoxygalactonojirimycin in Bombycis Faeces

    Institute of Scientific and Technical Information of China (English)

    QI Hui; ZHAO Hui; DING Zhen-ying; ZHU Yuan-yuan

    2011-01-01

    Objective To establish a simple and rapid method for the determination of 1,3-dideoxygalactonojirimycin in Bombycis Faeces, a potent glucosidase inihibitor, by HPLC. Methods A RP-HPLC method with fluorescence detection has been developed. Results The HPLC method developed in this research has a good reliability including accuracy and precision. The detection limit was less than 72 ng. Conclusion This method is sufficiently sensitive for determining 1,3-dideoxygalactonojirimycin in Bombycis Faeces and other related products.

  8. High performance liquid chromatography (HPLC fingerprints and primary structure identification of corn peptides by HPLC-diode array detection and HPLC-electrospray ionization tandem mass spectrometry

    Directory of Open Access Journals (Sweden)

    Chi Wang

    2016-01-01

    Full Text Available Corn peptides (CPs are reported to have many biological functions, such as facilitating alcohol metabolism, antioxidation, antitumor, antihypertension, and hepatoprotection. To develop a method for quality control, the high-performance liquid chromatography (HPLC system was applied. Twenty-eight common peaks were found in all the CPs of corn samples from Enshi, China, based on which, a fingerprinting chromatogram was established for use in quality control in future research. Subsequently, the major chemical constituents of these common peaks were identified respectively using the HPLC-diode-array detection electrospray ionization tandem mass spectrometry (DAD-ESI-MS/MS system, and 48 peptide fractions were determined ultimately. This was the first time for the majority of these peptides to be reported, and many of them contained amino acids of glutamine (Q, L and A, which might play an important role in the exhibition of the bioactivities of CPs. Many peptides had a similar primary structure to the peptides which had been proven to be bioactive such as facilitating alcohol metabolism, scavenging free radicals, and inhibiting lipid peroxidation. This systematical analysis of the primary structure of CPs facilitated subsequent studies on the relationship between the structures and functions, and could accelerate holistic research on CPs.

  9. Automated precolumn derivatization procedures in HPLC for biomedical and clinical applications

    NARCIS (Netherlands)

    Wolf, Johannes Hendrik

    1992-01-01

    This thesis describes three automated precolumn derivatization procedures for the analysis of carboxylic group-containing compounds. After derivatization with a suitable label, the derivatives are separated on reversed-phashed HPLC and detected by fluorescence. ... Zie: Summary

  10. Automated precolumn derivatization procedures in HPLC for biomedical and clinical applications

    NARCIS (Netherlands)

    Wolf, Johannes Hendrik

    1992-01-01

    This thesis describes three automated precolumn derivatization procedures for the analysis of carboxylic group-containing compounds. After derivatization with a suitable label, the derivatives are separated on reversed-phashed HPLC and detected by fluorescence. ... Zie: Summary

  11. Development and optimization of the SPE procedure for determination of pharmaceuticals in water samples by HPLC-diode array detection.

    Science.gov (United States)

    Mutavdzić Pavlović, Dragana; Babić, Sandra; Dolar, Davor; Asperger, Danijela; Kosutić, Kresimir; Horvat, Alka J M; Kastelan-Macan, Marija

    2010-02-01

    This paper focuses on the investigation of different types of SPE sorbents for the preconcentration of eight veterinary pharmaceuticals from water samples. The pharmaceuticals studied were sulfamethazine, sulfadiazine, sulfaguanidine, trimethoprim, oxytetracycline, enrofloxacin, norfloxacin and penicillin G/procaine. Five different SPE materials (Strata-X, Strata-X-C, Strata SDB-L, Strata C8 and Strata C18) from Phenomenex were compared with Oasis HLB with a view to obtaining the best cartridges for all pharmaceuticals investigated. Extraction efficiency was determined by HPLC with diode array detection (DAD). HPLC-DAD separation and quantification of the selected pharmaceuticals were carried out under gradient elution by a binary mixture of 0.01 M oxalic acid and ACN based on cyano modified column (LiChrosphere 100 CN) from Merck. Strata-X provided the best results in the preconcentration of 100 mL water samples, yielding average pharmaceutical recoveries of higher than 90%, except for sulfaguanidine (76.1%). The developed Strata-X-HLPC-DAD method was validated and applied, for the efficient investigation of reverse osmosis/nanofiltration membranes and for the removal of these eight pharmaceuticals from the production wastewater samples. NF90 and XLE membranes were shown to be the best for the rejection of all investigated pharmaceuticals.

  12. Dual-sensitive probe 1-imidazole-2-(5-benzoacridine)-ethanone for the determination of amines in environmental water using HPLC with fluorescence detection and online atmospheric chemical ionization-mass spectrometry identification.

    Science.gov (United States)

    You, Jinmao; Song, Cuihua; Fu, Yanyan; Sun, Zhiwei; Xia, Lian; Li, Yulin; Suo, Yourui

    2010-01-15

    Dual-sensitive probe of 1-imidazole-2-(5-benzoacridine)-ethanone (IBAE) for the determination of free amines with fluorescence detection and online highly sensitive atmospheric chemical ionization-mass spectrometry identification (APCI-MS) has been developed. 2-(Benzoacridine)-5-acetic acid (BAAA) reacts with coupling agent N,N'-carbonyldiimidazole (CDI) to form a highly activated amide intermediate 1-imidazole-2-(5-benzoacridine)-ethanone (IBAE), which is dual-sensitive probe. The amide intermediate (IBAE) reacts preferably with amines in dimethylformamide (DMF) solvent to give the high yields of derivatives. IBAE-amine derivatives are not only sensitive to fluorescence but also to MS ionizable efficiency. The percent ionization delta values change from 0 to 57.32% in aqueous acetonitrile and from 0 to 62.14% in aqueous methanol. The relative standard deviations of the peak areas with fluorescence detection for each amine are fluorescence detection limits (at a signal-to-noise ratio of 3) are in the range of 0.15-0.50ng/ml. The online APCI-MS detection limits are in the range of 2.07-8.51ng/ml (at a signal-to-noise ratio of 3). Therefore, the facile IBAE intermediate derivatization allowed the development of a highly sensitive and specific method for the quantitative analysis of trace levels of amines in environmental water.

  13. Changes in color-related compounds in tomato fruit exocarp and mesocarp during ripening using HPLC-APcI+-mass Spectrometry

    OpenAIRE

    Carrillo-López, A.; Yahia, E.M.

    2012-01-01

    Tomato is an important agricultural crop world-wide. Their pigments are very important in many ways. They have been associated with health benefits such as lowering the risk of some chronic diseases. Quantification of chlorophylls by spectrophotometry and Identification of carotenoids using liquid chromatography coupled to mass spectrometry, and quantification by HPLC-DAD was carried out in the exocarp and mesocarp of tomato fruit during 6 different ripeness stages (mature-green, breakers, tu...

  14. The formin DAD domain plays dual roles in autoinhibition and actin nucleation.

    Science.gov (United States)

    Gould, Christopher J; Maiti, Sankar; Michelot, Alphée; Graziano, Brian R; Blanchoin, Laurent; Goode, Bruce L

    2011-03-08

    Formins are a large family of actin assembly-promoting proteins with many important biological roles. However, it has remained unclear how formins nucleate actin polymerization. All other nucleators are known to recruit actin monomers as a central part of their mechanisms. However, the actin-nucleating FH2 domain of formins lacks appreciable affinity for monomeric actin. Here, we found that yeast and mammalian formins bind actin monomers but that this activity requires their C-terminal DAD domains. Furthermore, we observed that the DAD works in concert with the FH2 to enhance nucleation without affecting the rate of filament elongation. We dissected this mechanism in mDia1, mapped nucleation activity to conserved residues in the DAD, and demonstrated that DAD roles in nucleation and autoinhibition are separable. Furthermore, DAD enhancement of nucleation was independent of contributions from the FH1 domain to nucleation. Together, our data show that (1) the DAD has dual functions in autoinhibition and nucleation; (2) the FH1, FH2, and DAD form a tripartite nucleation machine; and (3) formins nucleate by recruiting actin monomers and therefore are more similar to other nucleators than previously thought.

  15. Comparison of two uncertainty dressing methods: SAD VS DAD

    Science.gov (United States)

    Chardon, Jérémy; Mathevet, Thibault; Le-Lay, Matthieu; Gailhard, Joël

    2014-05-01

    Hydrological Ensemble Prediction Systems (HEPSs) allow a better representation of meteorological and hydrological forecast uncertainties and improve human expertise of hydrological forecasts. An operational HEPS has been developed at EDF (French Producer of Electricity) since 2008 and is being used since 2010 on a hundred of watersheds in France. Depending on the hydro-meteorological situation, streamflow forecasts could be issued on a daily basis and are used to help dam management operations during floods or dam works within the river. A part of this HEPS is characterized by a streamflow ensemble post-processing, where a large human expertise is solicited. The aim of post-processing methods is to achieve better overall performances, by dressing hydrological ensemble forecasts with hydrological model uncertainties. The present study compares two post-processing methods, which are based on a logarithmic representation of the residuals distribution of the Rainfall-Runoff (RR) model, based on "perfect" forcing forecasts - i.e. forecasts with observed meteorological variables as inputs. The only difference between the two post-processing methods lies in the sampling of the perfect forcing forecasts for the estimation of the residuals statistics: (i) a first method, referred here as Statistical Analogy Dressing (SAD) model and used for operational HEPS, estimates beforehand the statistics of the residuals by streamflow sub-samples of quantile class and lead-time, since RR model residuals are not homoscedastic. (ii) an alternative method, referred as Dynamical Analogy Dressing (DAD) model, estimates the statistics of the residuals using the N most similar perfect forcing forecasts. The selection of this N forecasts is based on streamflow range and variation. On a set of 20 watersheds used for operational forecasts, both models were evaluated with perfect forcing forecasts and with ensemble forecasts. Results show that both approaches ensure a good post-processing of

  16. Comparative study of fourteen alkaloids from Uncaria rhynchophylla hooks and leaves using HPLC-diode array detection-atmospheric pressure chemical ionization/MS method.

    Science.gov (United States)

    Qu, Jialin; Gong, Tianxing; Ma, Bin; Zhang, Lin; Kano, Yoshihiro; Yuan, Dan

    2012-01-01

    The purpose of the study is to compare alkaloid profile of Uncaria rhynchophylla hooks and leaves. Ten oxindole alkaloids and four glycosidic indole alkaloids were identified using HPLC-diode array detection (DAD) or LC-atmospheric pressure chemical ionization (APCI)-MS method, and a HPLC-UV method for simultaneous quantification of major alkaloids was validated. The hooks are characterized by high levels of four oxindole alkaloids rhynchophylline (R), isorhynchophylline (IR), corynoxeine (C) and isocorynoxeine (IC), while the leaves contained high level of two glycosidic indole alkaloids vincoside lactam (VL) and strictosidine (S). The presented methods have proven its usefulness in chemical characterization of U. rhynchophylla hooks and leaves.

  17. Comprehensive Phenolic Profiling of Cyclopia genistoides (L. Vent. by LC-DAD-MS and -MS/MS Reveals Novel Xanthone and Benzophenone Constituents

    Directory of Open Access Journals (Sweden)

    Theresa Beelders

    2014-08-01

    Full Text Available A high-performance liquid chromatographic (HPLC method coupled with diode-array detection (DAD was optimized for the qualitative analysis of aqueous extracts of Cyclopia genistoides. Comprehensive insight into the phenolic profile of unfermented and fermented sample extracts was achieved with the identification of ten compounds based on comparison with authentic reference standards and the tentative identification of 30 additional compounds by means of electrospray ionization mass spectrometry (ESI-MS and tandem MS detection. Three iriflophenone-di-O,C-hexoside isomers, three xanthone-dihydrochalcone derivatives and one dihydrochalcone are herein tentatively identified for the first time in C. genistoides. Of special interest is one iriflophenone-di-O,C-hexoside present in large amounts. New compounds (tentatively identified for the first time in this species, and also in the genus Cyclopia, include two aromatic amino acids, one flavone, an iriflophenone-di-C-hexoside, a maclurin-di-O,C-hexoside, two tetrahydroxyxanthone-C-hexoside isomers, a tetrahydroxyxanthone-di-O,C-hexoside, two symmetric tetrahydroxyxanthone-C-hexoside dimers, nine glycosylated flavanone derivatives and five glycosylated phenolic acid derivatives. The presence of new compound subclasses in Cyclopia, namely aromatic amino acids and glycosylated phenolic acids, was demonstrated. The HPLC-DAD method was successfully validated and applied to the quantitative analysis of the paired sample extracts. In-depth analysis of the chemical composition of C. genistoides hot water extracts gave a better understanding of the chemistry of this species that will guide further research into its medicinal properties and potential uses.

  18. Sleepovers with Dad Can Be a Win-Win After Divorce

    Science.gov (United States)

    ... of thing happens when divorced dads have overnight parenting time," he said. Mothers also benefit, he said, because it gives them some relief from being a single, full-time parent. "Good quality relationships with parents ...

  19. Fluorescence Microscopy and HPLC Assay for Rapid Detection of Distribution and Content of Resveratrol in Polygonum cuspidatum%荧光显微技术与HPLC联用法快速检测虎杖组织器官中白藜芦醇的分布与含量

    Institute of Scientific and Technical Information of China (English)

    卜晓英; 董爱文; 管琼玉; 吴锋

    2012-01-01

    目的:建立荧光显微技术与HPLC法联用,快速检测不同生长期虎杖组织器官中白黎芦醇的分布与含量.方法:采用序贯试验设计冷冻切片的条件,荧光显微镜观察;超声波辅助提取所切材料中的白黎芦醇,HPLC检测其含量.结果:冷冻切片的条件为阿拉伯胶浓度先20%后40%,浸渍时间20%为5~6h、40%为2~5 min,冷冻室温度-5℃,厚度为15μm;白藜芦醇主要积累在虎杖各器官、组织与细胞的纤维和纤维素中,其含量在根状茎中髓>木质部>韧皮部>周皮;器官间则是根状茎的芽>根状茎>地上茎>叶;根茎中白藜芦醇的含量随年龄增长而增加.结论:荧光显微镜观察结果与HPLC检测结果一致,说明该方法简便快捷,结果可靠,为虎杖最佳采收期与收购品质的确定提供了快速可靠的检测方法.%Objective:To establish fluorescence microscopy combined with HPLC method for rapid detection the distribution and content of resveratrol tissues in different growth stages of Polygonum cuspidatum. Methods; Used sequential experiment to design conditions of frozen and observe of the section by fluorescence microscopy; Resveratrol was extracted by ultrasonic-assisted extraction and its content was detected by HPLC. Results:The results showed that frozen condition for concentration of gum Arabic was from 20% (dipping time was 5 ~6 h) to 40% (2 ~5min) ,the freezer temperature was -5 ℃ ,and the thickness was 15 μm. Resveratrol in polygonum cuspidatum was mainly accumulated in the organs, tissues and cells of fiber and cellulose, its content in rhizomes declined as the following sequence:spinal cord >xylem > phloem > periderm;Its content declined in organ as the following sequence; buds > rhizomes > ground stem > leaves; The content of resveratrol in root increased with age. Conclusion: The results of fluorescence microscopic observation is in accordance with the HPLC results,indicating that the method is

  20. Microwave rapid extraction of free fatty acids from Zanthoxylum bungeanum Maxim and their HPLC fluorescence detection by pre-column derivatization%花椒游离脂肪酸的微波快速提取与衍生化HPLC荧光检测方法研究

    Institute of Scientific and Technical Information of China (English)

    郑振佳; 席兴军; 赵先恩; 赵镭; 张璐璐; 郑佳佳

    2013-01-01

    建立了花椒中游离脂肪酸的微波快速提取技术,并采用苯并[b]吖啶酮-5-乙基对甲苯磺酸酯(BAETS)作为柱前衍生试剂,建立了19种长链饱和与不饱和游离脂肪酸的衍生化HPLC荧光检测分析方法.微波提取功率700W,溶剂为体积比1∶1的氯仿/甲醇,70℃提取10min,可达近100%提取率.采用色谱柱Eclipse XDB-C8(4.6mm× 150mm,5μm),酸性乙腈/水流动相进行梯度洗脱,HPLC荧光激发和发射波长分别为273nm和505nm.同时进行在线质谱辅助鉴定(APCI-MS),更提高了HPLC荧光定量的准确度.结果表明,微波提取快速高效,分析方法线性良好,线性范围14.286~0.0139μmol/L,最低检出限为0.636nmol/L,回收率在87.60%~ 106.59%之间.本方法快速、准确、高灵敏,可作为花椒相关样品的标准化检测方法.%Free fatty acids (FFAs) in Zanthoxylum bungeanum Maxim were extracted by microwave-assisted extraction and derivatized using 2-(12-oxobenzo[b]acridin-5 (12H)-yl) ethyl-4-toluenesulfonate (BAETS) as derivatization reagent.This method was proposed to detect 19 long-chain saturated and unsaturated FFA derivatives by HPLC fluorescence detection using pre-column derivatization.Microwave-assisted extract percentages almost reached 100% when FFAs were extracted at microwave power 700W,70℃ and 10min using chloroform/methanol(1∶1,v∶v).Good chromatographic separation was achieved using acidic acetonitrile/ water as mobile phase on Eclipse XDB-C8 column(4.6mm×150mm,5μm) by gradient elution.On-line APCI-MS identification was used for producing more accurate HPLC fluorescence quantitation.Fluorescence excitation and emission wavelengths for HPLC detection were 273nm and 505nm,respectively.The results indicated that microwave-assisted extraction was fast and efficient,and the developed analytical method had good linearity.Linear range was from 14.286 to 0.0139μmol/L,the lowest detection limits was 0.636nmol/L,the recovery was in the range of 87

  1. [DAD-6: an abbreviated version of the DAD scale (disability assessment for dementia). An instrument for detection of loss of autonomy at an early stage].

    Science.gov (United States)

    de Rotrou, Jocelyne; Wu, Ya-Huei; Djabelkhir, Leila; Seux, Marie-Laure; Hugonot, Laurence; Rigaud, Anne-Sophie; Hanon, Olivier; Vidal, Jean-Sébastien

    2014-09-01

    This paper presents the French version of DAD-6, a validated instrument for the assessment of IADL (instrumental activities of daily living) considered as intentional and complex activities. A loss of autonomy remains a major criterion in the diagnosis of dementia. In addition, IADL assessment is recommended as a primary outcome in dementia drug trials. Since the publication in 1969 by Lawton and Brody of an IADL scale, many instruments have been developed. However, their psychometric properties remain to be improved. The need for improving the early diagnosis yielded to the design of DAD-6, an instrument allowing capturing subtle difficulties in IADL management. The DAD-6 scale emphasizes the role of the cognitive function, mainly the executive function in early IADL impairment. DAD-6 requires the participation of an informant (a patient's proxy). Relative to patients' self-reports or performance-based methods, informant-based questionnaires are the most common and practical methods used in memory clinics. In previous work, DAD-6 score gradually decreased with increasing severity of the cognitive status. The present work shows the inter-rater reliability of DAD-6. The use of the scale with the same informants by one neurologist and two neuropsychologists, separately, indicated a high agreement between raters (alpha of Krippendorff>0.80).This work also highlights the main sources of bias in the context of evaluation based on subjective judgement. The authors stress the necessity of: 1--a clarification of the relationship between cognitive function and IADL; 2--the measurement of IADL performance in a routine neuropsychological assessment by experienced professionals.

  2. Determination and characterization of cysteine, glutathione and phytochelatins (PC₂₋₆) in Lolium perenne L. exposed to Cd stress under ambient and elevated carbon dioxide using HPLC with fluorescence detection.

    Science.gov (United States)

    Ju, Xue Hai; Tang, Shirong; Jia, Yan; Guo, Junkang; Ding, Yongzhen; Song, Zhengguo; Zhao, Yujie

    2011-06-15

    Metal-binding thiols, involved in detoxification mechanisms in plant and other organism under heavy metal stress, are receiving more and more attentions, and various methods have been developed to determine related thiols such as cysteine (Cys), glutathione (GSH) and phytochelatins (PCs). In present study, an HPLC method was established for simultaneous determination of Cys GSH and PC(2-6) after treatment with disulfide reductant of tris (2-carboxyethyl) phosphine hydrochloride (TCEP) and thiolyte reagent of monobromobimane (mBBr). The separation of thiol derivatives was performed on an Agilent Zorbax Eclipse XDB-C18 column (4.6 mm × 30 mm, 1.8 μm) with a linear gradient elution of 0.1% (v/v) trifluoroacetic acid (TFA)-acetonitrile (ACN) at 0.8 mL min(-1). The temperature of the column was maintained at 25°C. The excitation and emission wavelengths were set at 380 and 470 nm, respectively. The thiol derivatives were well separated in 19 min, and the total analysis time was 30 min. The established method was proved selective, specific and reproducible, and could be applicable to determine Cys, GSH and PC(2-6) and to evaluate their roles in detoxification mechanisms in Cd-treated Lolium perenne L. under ambient and elevated carbon dioxide (CO(2)). It was found that the total SH contents and proportions of thiols in roots and shoots were dependent on Cd concentration, whereas the total SH contents decreased and the proportions of thiols altered without significance at elevated CO(2) level.

  3. Determination of Aliphatic Amines by Pre-column Derivatization with High-performance Liquid Chromatography (HPLC) by Fluorescence Detection%柱前衍生化HPLC荧光法测定脂肪胺化合物

    Institute of Scientific and Technical Information of China (English)

    刘素娟; 孙学军; 朱芳; 尤进茂

    2007-01-01

    采用新型荧光试剂4-(1-甲基-1H-菲并[9,10-d]咪唑-2-)苯甲酸(MPIBA)进行柱前衍生并经荧光检测对脂肪胺进行了高效液相色谱(HPLC)分离.衍生物的荧光激发和发射波长分别为λem=260 nm,λem=446 nm.80℃下在四氢呋喃(THF)溶剂中用N-乙基-N'-[(3-二甲氨基)丙基]碳二亚胺盐酸盐(EDC)作催化剂,衍生反应10min后获得稳定的荧光产物.在Eclipse XDB-C8色谱柱(4.6×150 mm,5 mm)上,采用梯度洗脱对12种游离脂肪胺衍生物进行了优化分离,实现了各种脂肪胺衍生物的快速、准确测定.该方法具有良好的重现性,多数脂肪胺的线性回归系数大于0.9996,检测限在10.5-53.4 fmol范围内,对实际样品臭豆腐中脂肪胺的测定测定结果满意.

  4. 脂肪胺荧光衍生物的高效液相色谱分离及质谱鉴定%HPLC Determination of Aliphatic Amines with Fluorescence Detection and Mass Spectrum Identification

    Institute of Scientific and Technical Information of China (English)

    赵先恩; 石运伟; 陈向明; 张海峰; 尤进茂

    2005-01-01

    采用新型荧光衍生试剂2-(9-吖啶酮)-乙酸(AAA)进行柱前衍生并经荧光检测对脂肪胺进行了高效液相色谱(HPLC)分离和在线质谱定性.衍生物荧光激发和发射波长为λex=404nm,λem=440 nm.30℃下在乙腈溶剂中用N-乙基-N'-[(3-二甲氨基)丙基]碳二亚胺盐酸盐(EDC)做催化剂,衍生反应20 min后获得稳定的荧光产物.在Hypersil BDS C18(4.6 mm×100mm,5μm)色谱柱上,采用梯度洗脱对12种脂肪胺衍生物进行了优化分离.采用大气压化学电离源(APCI Source)正离子模式进行在线柱后质谱定性,实现了各种脂肪胺衍生物的快速、准确测定.该方法具有良好的重现性,多数脂肪胺的线性回归系数大于0.999 6,检测限为12.09~25.52fmol.

  5. Profiling LC-DAD-ESI-TOF MS method for the determination of phenolic metabolites from avocado (Persea americana).

    Science.gov (United States)

    Hurtado-Fernández, Elena; Carrasco-Pancorbo, Alegría; Fernández-Gutiérrez, Alberto

    2011-03-23

    A powerful HPLC-DAD-ESI-TOF MS method was established for the efficient identification of the chemical constituents in the methanolic extracts of avocado (Persea americana). Separation and detection conditions were optimized by using a standard mix containing 39 compounds belonging to phenolic acids and different categories of flavonoids, analytes that could be potentially present in the avocado extracts. Optimum LC separation was achieved on a Zorbax Eclipse Plus C18 analytical column (4.6×150 mm, 1.8 μm particle size) by gradient elution with water+acetic acid (0.5%) and acetonitrile as mobile phases, at a flow rate of 1.6 mL/min. The detection was carried out by ultraviolet-visible absorption and ESI-TOF MS. The developed method was applied to the study of 3 different varieties of avocado, and 17 compounds were unequivocally identified with standards. Moreover, around 25 analytes were tentatively identified by taking into account the accuracy and isotopic information provided by TOF MS.

  6. Simultaneous analysis of carbohydrates and organic acids by HPLC-DAD-RI for monitoring goat's milk yogurts fermentation.

    Science.gov (United States)

    da Costa, Marion Pereira; Frasao, Beatriz da Silva; Lima, Bruno Reis Carneiro da Costa; Rodrigues, Bruna Leal; Conte Junior, Carlos Adam

    2016-05-15

    During yogurt manufacture, the lactose fermentation and organic acid production can be used to monitor the fermentation process by starter cultures and probiotic bacteria. In the present work, a simple, sensitive and reproducible high-performance liquid chromatography with dual detectors, diode array detector and refractive index was validated by simultaneous analysis of carbohydrates and organic acids in goat milk yogurts. In addition, pH and bacterial analysis were performed. Separation of all the compounds was performed on an Aminex HPX-87H column (300×7.8 mm, 9 µm) utilizing a 3 mmol L(-1) sulfuric acid aqueous mobile phase under isocratic conditions. Lactose, glucose, galactose, citric, lactic and formic acids were used to evaluate the following performance parameters: selectivity, linearity, precision, limit of detection (LOD), limit of quantification (LOQ), decision limits (CCα), detection capabilities (CCβ), recovery and robustness. For the method application a six goat milk yogurts were elaborated: natural, probiotic, prebiotic, symbiotic, cupuassu fruit pulp, and probiotic with cupuassu fruit pulp. The validated method presented an excellent selectivity with no significant matrix effect, and a broad linear study range with coefficients of determination higher than 0.995. The relative standard deviation was lower than 10% under repeatability and within-laboratory reproducibility conditions for the studied analytes. The LOD of the method was defined from 0.001 to 0.003 µg g(-1), and the LOQ from 0.003 to 0.013 µg g(-1). The CCα was ranged from 0.032 to 0.943 µg g(-1), and the CCβ from 0.053 to 1.604 µg g(-1). The obtained recovery values were from 78% to 119%. In addition, the method exhibited an appropriate robustness for all parameter evaluated. Base in our data, it was concluded that the performance parameters demonstrated total method adequacy for the detection and quantification of carbohydrates and organic acids in goat milk yogurts. The application of the method was successfully applied to monitoring different goat milk yogurts during fermentation.

  7. Determination of haloperidol in biological samples using molecular imprinted polymer nanoparticles followed by HPLC-DAD detection.

    Science.gov (United States)

    Ebrahimzadeh, Homeira; Dehghani, Zahra; Asgharinezhad, Ali Akbar; Shekari, Nafiseh; Molaei, Karam

    2013-09-10

    In this study an extraction procedure using molecular imprinted polymer nanoparticles for the determination of haloperidol in biological samples is proposed. The haloperidol imprinted polymer nanoparticles were synthesized successfully by precipitation polymerization in a flask containing haloperidol as a template, ethyleneglycoldimethacrylate as a crosslinking agent, methacrylic acid as a functional monomer, and 2,2'-azobisisobutyronitrile as an initiator. The leached and unleached polymer nanoparticles have been characterized by infrared spectroscopy and scanning electron microscopy. The effect of different variables such as the pH of solution, uptake and elution time, type, and the least amount of eluent for elution of haloperidol from polymer was evaluated. Extraction efficiencies more than 97% were obtained by elution of the polymer with 1.5 mL of methanol-acetic acid-trifluoroacetic acid 79.9:20:0.1. Under optimal conditions maximum adsorption capacity was obtained 153.84 mg g(-1). The detection limit of the proposed procedure was between 0.2 and 0.35 μg L(-1). Finally this method was applied to the determination of haloperidol in plasma and urine samples and satisfactory results were achieved (RSD<6.9%).

  8. Simultaneous, stability indicating, HPLC-DAD determination of guaifenesin and methyl and propyl-parabens in cough syrup.

    Science.gov (United States)

    Grosa, Giorgio; Del Grosso, Erika; Russo, Roberta; Allegrone, Gianna

    2006-06-07

    A stability indicating high performance liquid chromatography procedure has been developed for the simultaneous determination of guaifenesin (GUA), methyl p-hydroxybenzoate (MHB) and propyl p-hydroxybenzoate (PHB) in a commercial cough syrup dosage form. The method was specific and stability indicating as chromatographic conditions were selected to provide adequate separation of GUA, MHB and PHB from the putative degradation products guaiacol (GUAI) and p-hydroxybenzoic acid (HBA) as well as from excipients. The isocratic separation and quantitation were achieved within 17 min on a 150-mm column with an ether-linked phenyl stationary phase and a hydrophilic endcapping. The mobile phase was constituted of eluant A: aqueous phosphate buffer (pH 3.0, 10 mM)/acetonitrile 25/75 (v/v) and eluant B:methanol; the A:B ratio was 85:15 (v/v) with a flow rate 1 ml min-1 and detection of analytes at 254 and 276 nm. The method showed good linearity for the GUA-MHB-PHB mixture in the 95-285, 4-12, and 1-3 microg ml-1 ranges, respectively, being all the square of the correlation coefficients greater than 0.999. The interday R.S.D.s were 1.17, 1.14, and 0.91%, for GUA, MHB, and PHP, respectively. The method demonstrated also to be accurate; indeed the average recoveries, at 100% of the target assay concentration, were 100.5, 100.3, and 100.7% with relative standard deviations of 0.8, 0.7, and 0.4% for GUA, MHB, and PHB, respectively, from laboratory prepared samples. The applicability of the method was evaluated in commercial dosage form analysis as well as in stability studies.

  9. Chemical Characteristics Combined with Bioactivity for Comprehensive Evaluation of Tumuxiang Based on HPLC-DAD and Multivariate Statistical Methods

    Directory of Open Access Journals (Sweden)

    Xia Gao

    2016-08-01

    Full Text Available Background: The dried roots of Inula helenium L. (IH and Inula racemosa Hook f. (IR are used commonly as folk medicine under the name of “tumuxiang (TMX”. Phenolic acid compounds and their derivatives, as main active constituents in IH and IR, exhibit prominent anti-inflammation effect.

  10. Forensic Investigation of Formaldehyde in Illicit Products for Hair Treatment by DAD-HPLC: A Case Study.

    Science.gov (United States)

    Oiye, Erica N; Ribeiro, Maria Fernanda M; Okumura, Leonardo L; Saczk, Adelir A; Ciancaglini, Pietro; de Oliveira, Marcelo F

    2016-07-01

    The illegal use of formalin (commercial formaldehyde) in cosmetic products harms the health of individuals exposed to this substance. Over the last years, the commercial availability of these products, especially those containing irregular dosage of formaldehyde, has increased in Brazil. This work analyzes some products for hair treatment available in the Brazilian market and verifies their safety. The adopted analytical methodology involved sample derivatization with 2,4-dinitrophenylhydrazine, followed by high-performance liquid chromatography with ultraviolet detection (UV-VIS) at λ = 365 nm. The limit of quantification is 2.5 × 10(-3%) w/w, and the recovery tests were around 93%. Some of the samples contained high and illegal formaldehyde levels ranging from 9% to 19% (w/w) and others presented suitable concentrations of the analyte. On the basis of the results, this work discusses the efficiency and practicality of this analytical method for forensic purposes. © 2016 American Academy of Forensic Sciences.

  11. Study on pharmacokinetics and tissue distribution of the isocorydine derivative (AICD) in rats by HPLC-DAD method.

    Science.gov (United States)

    Chen, Yali; Yan, Qian; Zhong, Mei; Zhao, Quanyi; Liu, Junxi; Di, Duolong; Liu, Jinxia

    2015-05-01

    A simple and effective high-performance liquid chromatography with diode-array detection method coupled with a liquid-liquid extraction pretreatment has been developed for determining the pharmacokinetics and tissue distribution of a novel structurally modified derivative (8-acetamino-isocorydine) of isocorydine. According to the in vivo experiments data calculations by DAS 2.0 software, a two-compartment metabolic model was suitable for describing the pharmacokinetic of 8-acetamino-isocorydine in rats. 8-Acetamino-isocorydine was absorbed well after oral administration, and the absolute bioavailability was 76.5%. The half-life of 8-acetamino-isocorydine after intravenous and oral administration was 2.2 h and 2.0 h, respectively. In vivo, 8-acetamino-isocorydine was highly distributed in the lungs, kidney and liver; however, relatively little entered the brain, suggesting that 8-acetamino-isocorydine could not easily pass through the blood brain barrier. Our work describes the first characterization of the pharmacokinetic parameters and tissue distribution of 8-acetamino-isocorydine. The acquired data will provide useful information for the in vivo pharmacology of 8-acetamino-isocorydine, and can be applied to new drug research.

  12. Study on pharmacokinetics and tissue distribution of the isocorydine derivative (AICD in rats by HPLC-DAD method

    Directory of Open Access Journals (Sweden)

    Yali Chen

    2015-05-01

    Full Text Available A simple and effective high-performance liquid chromatography with diode-array detection method coupled with a liquid-liquid extraction pretreatment has been developed for determining the pharmacokinetics and tissue distribution of a novel structurally modified derivative (8-acetamino-isocorydine of isocorydine. According to the in vivo experiments data calculations by DAS 2.0 software, a two-compartment metabolic model was suitable for describing the pharmacokinetic of 8-acetamino-isocorydine in rats. 8-Acetamino-isocorydine was absorbed well after oral administration, and the absolute bioavailability was 76.5%. The half-life of 8-acetamino-isocorydine after intravenous and oral administration was 2.2 h and 2.0 h, respectively. In vivo, 8-acetamino-isocorydine was highly distributed in the lungs, kidney and liver; however, relatively little entered the brain, suggesting that 8-acetamino-isocorydine could not easily pass through the blood brain barrier. Our work describes the first characterization of the pharmacokinetic parameters and tissue distribution of 8-acetamino-isocorydine. The acquired data will provide useful information for the in vivo pharmacology of 8-acetamino-isocorydine, and can be applied to new drug research.

  13. Statistical Optimization of Evaporative Light Scattering Detection for Molten Sucrose Octaacetate and Comparison With Ultraviolet Diode Array Detection Validation Parameters Using Tandem HPLC Ultraviolet Diode Array Detection/Evaporative Light Scattering Detection-Specific Stability-Indicating Method.

    Science.gov (United States)

    Parmar, Rudrangi; Ghanta, Ajay; Haware, Rahul V; Johnson, Paul R; Stagner, William C

    2016-12-01

    A sucrose octaacetate (SOA) gradient HPLC evaporative light scattering detection (ELSD) and low-wavelength UV-diode array detection (UV-DAD)-specific stability-indicating method development and validation comparison is reported. A central composite response surface design and multicriteria optimization was used to maximize molten SOA area-under-the-curve response and signal-to-noise ratio. The ELSD data were also analyzed using multivariate principal component analysis, analysis of variance, and standard least squares effects modeling. The method suitability and validation parameters of both methods were compared. To the authors' knowledge, this is the first report that validates an ELSD method using a molten analyte. SOA exhibited a low molar absorptivity of 439 absorption units/cm/M in water at 210 nm requiring low-wavelength UV-DAD detection. The low-wavelength UV-DAD method provided substantially better intraday and interday precision, intraday and interday goodness-of-fit, detection limit, and quantitation limit than ELSD. ELSD exhibited a 60-fold greater area-under-the-curve response, better resolution, and 58% more theoretical plates. On balance, the UV-DAD method was chosen for SOA chemical kinetic studies. This study illustrates that ELSD may not always be the best alternative to gradient HPLC low-wavelength UV detection. Copyright © 2016. Published by Elsevier Inc.

  14. Does digit ratio (2DAD) predict penile length?

    Institute of Scientific and Technical Information of China (English)

    Denise Brooks McQuade

    2011-01-01

    @@ Now advertized in at least one locale as the 'sexy ratio' with the caption,'Your hands give away your hotness' (http://io9.com/#!5794008; accessed 3 May 2011),an online summary of a recent study linking facial attractiveness to the ratio of the index and ring fingers1 indicates that digit ratio (2DAD)research has reached mainstream culture.Digit ratio,most commonly the ratio of the index to ring fingers in humans,is sexually dimorphic (males have lower values than females) with greater differences on their right hand 2s Ratios are determined early in development and remain relatively stable thereafter.4 Perhaps the popular appeal is the relative simplicity of the measure,and the fact that researchers interested in any number of behavioral or physiological traits correlatedwith hormone activity have adopted digit ratio as a convenient biomarker for prenatal androgen exposure.5 Over the past decade,the correlation of digit ratio with sexual behavior and other aspects of reproductive biology has been well documented6-9 and there is a growing list of traits with links to digit ratio,although the associations are less well established.

  15. Development and validation of an ultra-high performance LC-MS/MS assay for intracellular SN-38 in human solid tumour cell lines: comparison with a validated HPLC-fluorescence method.

    Science.gov (United States)

    Ghazaly, Essam; Perry, Jackie; Kitromilidou, Christiana; Powles, Thomas; Joel, Simon

    2014-10-15

    A simple and rapid ultra-high performance liquid chromatography-mass spectrometry/mass spectrometry (UPLC-MS/MS) method has been developed for measuring intracellular concentrations of the anticancer agent 7-ethyl-10-hydroxycamptothecin (SN-38) in tumour cells using camptothecin (CPT) as internal standard. SN-38 extraction was carried out using acidified acetonitrile. SN-38 and CPT were separated on a PFP column using gradient elution with acidified water and acetonitrile. SN-38 and CPT were quantified using a triple quadrupole mass spectrometry system. Least square regression calibration lines were obtained with average correlation coefficients of R(2)=0.9993±0.0016. The lower limit of detection (LOD) and lower limit of quantification (LOQ) for SN-38 were 0.1 and 0.3ng/ml, respectively. CPT recovery was 98.5±13% and SN-38 recoveries at low quality control (LQC, 5ng/ml) and high quality control (HQC, 500ng/ml) were 89±6% and 95±8%, respectively. The intra- and inter-day imprecision for LQC was 5.8 and 8.5%, and for HQC was 6.3 and 4.4%, respectively. The method was compared to a validated high performance liquid chromatography-fluorescent method. In addition, the method has been successfully applied to determine the intracellular accumulation of SN-38 investigating the transport through ABCB1 (P-gp) and ABCG2 (BCRP) efflux pumps in colorectal cancer cell lines.

  16. HPLC: Early and Recent Perspectives.

    Science.gov (United States)

    Karger, Barry L.

    1997-01-01

    Provides a perspective on what it was like in the early days of high-performance liquid chromatography (HPLC) and several of the key developments. Focuses on the advances in HPLC generally, and more specifically for the biological sciences, that were necessary for the method to reach the preeminent stage of today. Contains 20 references. (JRH)

  17. HPLC: Early and Recent Perspectives.

    Science.gov (United States)

    Karger, Barry L.

    1997-01-01

    Provides a perspective on what it was like in the early days of high-performance liquid chromatography (HPLC) and several of the key developments. Focuses on the advances in HPLC generally, and more specifically for the biological sciences, that were necessary for the method to reach the preeminent stage of today. Contains 20 references. (JRH)

  18. [Degradation of halogenated compounds by haloalkane dehalogenase DadA from Alcanivorax dieselolei B-5 ].

    Science.gov (United States)

    Li, Anzhang; Shao, Zongze

    2014-09-04

    [OBJECTIVE] Alcanivorax dieselolei B-5 is an important oil-degrading bacterium. We studied its substrate range and degradation of halogenated compounds. [METHODS] Growth capability of B-5 was examined with different halogenated substrates as sole carbon source. A putative haloalkane dehalogenase (HLD) gene named dadA was found from the genome of strain B-5 and analyzed by sequence alignment, phylogenetic analysis and homologous modeling. After heterologous expression in Escherichia coli and purification, the activity of DadA towards 46 substrates was determined. [RESULTS] Strain B-5 was capable of utilizing various halogenated compounds (C3-C,8) as the sole carbon source. DadA had typical catalytic pentad residues of HLD-II subfamily, but it was independent from other members of this subfamily according to phylogenetic analysis. Activity assay showed that DadA has higher specificity and narrower substrate range than other characterized HLDs and it only showed activity toward 1,2,3-tribromopropane, 1,2-dibromo-3-chloropropane and 2,3-dichloroprop-1-ene among 46 tested substrates. [CONCLUSIONS] Strain B-5 and its HLD DadA can degrade halogenated aliphatic pollutants although.

  19. Effect of Probiotic Lactobacillus sp. Dad13 on Humoral Immune Response of Balb/C Mice Infected with Salmonella typhimurium

    Directory of Open Access Journals (Sweden)

    Ika Dyah Kusumawati

    2015-10-01

    Full Text Available An indigenous strain of lactic acid bacterium (LAB identified as Lactobacillus spp. Dad13 (Dad13, isolatedfrom traditional fermented buffalo milk, was found to be potential as probiotic. The aim of this research was to studythe effect of probiotic Dad13 on humoral immune response of Balb/C mice infected with Salmonella typhimurium. Thespecific objective was to find out the effect of different Dad13 consumption time (before and along with infection of S.typhimurium on the humoral immune response of Balb/C mice. The experiment was conducted by in vivo trial on 20males of Balb/C mice, age of 6-8 weeks, fed with AIN-93 standard diet. The mice were assigned into 4 groups. Eachgroup received the following treatments, ie. Dad13 only, Dad13 before infection, Dad13 along with infection andSalmonella infection only. A volume of 100 μl Dad13 cell suspensions (1010 CFU/ml were given by oral forced feedingdaily for a week, at week 3 for group before infection and at week 4 for group of Dad13 only and Dad13 along withinfection. Salmonella infection (109 CFU/ml was given once orally at week 4 to all groups except group treated withDad13 only. The humoral immune response of Balb/C mice was detected 2 weeks after infection by measuring thetiters of IgG and IgA specific from serum and mucosal intestinal liquid samples using Enzyme-linked ImmunosorbentAssay (ELISA method. The result indicated that humoral immune response of Balb/C mice consuming Dad13 beforeand along with Salmonella infection were significantly different (p<0.05. Dad13 consumption along with Salmonellainfection increased circulated IgG and IgA as well as secretory IgA. It can be concluded that Dad13 probiotic feedingalong with infection increased humoral immune response more significantly compared to that before infection.Key words : Probiotic, Lactobacillus sp. Dad13, Immune response, Salmonella typhimurium

  20. Isocratic LC-DAD-FLD method for the determination of flavonoids in paprika samples by using a rapid resolution column and post-column pH change.

    Science.gov (United States)

    Monago-Maraña, Olga; Muñoz de la Peña, Arsenio; Galeano-Díaz, Teresa

    2016-05-15

    The determination of flavonoid compounds in paprika samples has been performed by liquid chromatography in series diode array and fluorescence detection (LC-DAD-FLD), by means of a pH change to basic medium just before FLD detection. The validation of the method was performed through the establishment of the external standard calibration curves and the analytical figures of merit. Limits of detection ranging from 0.006 to 0.02 mg L(-1) and 0.007 to 0.09 mg L(-1) were achieved using DAD and FLD detection, respectively. The experimental conditions to carry out the hydrolysis procedure to obtain flavonoid aglycones from flavonoid glycosides have been optimized applying an experimental design and the response surface methodology. The final conditions selected were 2.5M HCl during 45 min at 85°C. The repeatability of this procedure was assayed and relative standard deviation (RSD) values for concentration of quercetin and luteolin compounds were lower than 2%. The quantification of quercetin, luteolin and kaempferol compounds was carried out in less than 6 min in paprika samples by means of the external standard calibration. The analytes were extracted with methanol and the extracts were previously subjected to a cleanup procedure to extend the use of the chromatographic column.

  1. Online screening of nitric oxide scavengers in natural products using high performance liquid chromatography coupled with tandem diode array and fluorescence detection.

    Science.gov (United States)

    Li, Dapeng; Wang, Ting; Guo, Yujie; Hu, Yuanjia; Yu, Boyang; Qi, Jin

    2015-12-18

    Nitric oxide (NO) is an important cellular signaling molecule with extensive physiological and pathophysiological effects. NO scavengers have the potential to treat inflammation, septic shock and other related diseases, and numerous examples have been chemically synthesized or isolated from natural products. The chemical diversity of natural products, however, means that a huge effort is necessary to efficiently screen and identify bioactive compounds, especially NO scavengers. In this article, we propose an effective analytical method to screen for NO scavengers in three natural products using an online system that couples high performance liquid chromatography with tandem diode array and fluorescence detection (HPLC-DAD-FLD). Eighteen compounds from radix of Scutellaria baicalensis Georgi and green tea displayed significant NO scavenging activity whereas components of Pueraria lobata (Willd.) Ohwi had no discernable activity. The structures of the active compounds were elucidated using Agilent Accurate-Mass Q-TOF LC/MS system. Preliminary analysis of structure-activity relationships indicated that, in flavonoids, a 2,3-double bond and a 3-H atom or a 3-OH group are essential for activity. In tannins, poly-hydroxyl groups are important for NO scavenging activity. Method validation indicated that the newly developed method is both reliable and repeatable. The online method that we present provides a simple, rapid and effective way to identify and characterize NO scavengers present in natural products.

  2. Determination of plasma total homocysteine and cysteine using HPLC with fluorescence detection and an ammonium 7-fluoro-2, 1, 3-benzoxadiazole-4-sulphonate (SBD-F) derivatization protocol optimized for antioxidant concentration, derivatization reagent concentration, temperature and matrix pH.

    Science.gov (United States)

    Daskalakis, I; Lucock, M D; Anderson, A; Wild, J; Schorah, C J; Levene, M I

    1996-01-01

    A sensitive HPLC-fluorescence method for determining total endogenous plasma homocysteine (Hcy), cysteine (Cys) and cysteinylglycine (Cys-Gly) following derivatization with ammonium 7-fluoro 2,1,3-benzoxadiazole-4-sulphonate (SBD-F) is described. Quantitation utilizes an internal standard, 2-mercaptoethylamine. The derivatization procedure has been optimized for concentration of SBD-F, reducing agent (tributylphosphine) and temperature. Findings indicate that values for plasma determinations vary according to the nature of the matrix in which calibration standards are made up. If quantitation is based on a peak height ratio, then standards should be made up in either pH 7.4 phosphate buffered saline or plasma taking into account the endogenous thiol concentration. These findings are based on calibration data, and 30 plasma samples quantified using thiol standards made up in plasma, pH 7.4 and pH 9.5 buffers. By defining how this matrix/pH effect influences thiol quantitation, it should be possible to make a more meaningful comparison of Hcy measurements between laboratories. The chromatographic separation was investigated at several mobile-phase pH values with the following conditions ascertained to be optimal: a mobile phase consisting of 5% (v/v) acetonitrile in 0.1 M KH2PO4, pH 2.15 was run at a flow rate of 0.5 mL/min. It was used in conjunction with a Supelco LC-18 base deactivated analytical column (150 x 4.6 cm i.d. 3 microM bonded silica). The internal standard and thiols were measured by fluorescence detection at 385 nm excitation and 515 nm emmission. Plasma levels are easily measured in a 100 microL volume. Storage for 2 months at -20 degrees C resulted in no deterioration of thiols. Furthermore, no difference in thiol levels was observed between bloods collected in lithium heparin and EDTA. Collected blood should, however, be separated as soon as possible to avoid red cell metabolism of Hcy which was observed in a case of hyperhomocysteinemia. Once

  3. Fluorescent compounds present in food

    OpenAIRE

    Soto Serrano, Axel

    2014-01-01

    Póster The food industry demands fast, reliable, cheap and reproducible methods for quality and process control. This bibliographic review work investigates florescence spectroscopy, a method that couldn’t be used in food until the recent technological advances, concretely front-face fluorescence and chemometric tools. This technology presents advantages as compared to classical methods like HPLC or capillary electrophoresis, which require qualified staff, sample preparation and are time-c...

  4. Regulation and Characterization of the dadRAX Locus for d-Amino Acid Catabolism in Pseudomonas aeruginosa PAO1▿

    OpenAIRE

    2011-01-01

    d-Amino acids are essential components for bacterial peptidoglycan, and these natural compounds are also involved in cell wall remodeling and biofilm disassembling. In Pseudomonas aeruginosa, the dadAX operon, encoding the d-amino acid dehydrogenase DadA and the amino acid racemase DadX, is essential for d- and l-Ala catabolism, and its expression requires a transcriptional regulator, DadR. In this study, purified recombinant DadA alone was sufficient to demonstrate the proposed enzymatic act...

  5. Identification of Radical Scavenging Compounds in Rhaponticum carthamoides by Means of LC-DAD-SPE-NMR

    NARCIS (Netherlands)

    Miliauskas, G.; Beek, van T.A.; Waard, de P.; Venskutonis, R.P.; Sudhölter, E.J.R.

    2005-01-01

    A hyphenated LC-DAD-SPE-NMR setup in combination with on-line radical scavenging detection has been applied for the identification of radical scavenging compounds in extracts of Rhaponticum carthamoides. After NMR measurements, the pure compounds were infused into a mass spectrometer. The technique

  6. She Has Great Spirit: Insight into Relationships between American Indian Dads and Daughters

    Science.gov (United States)

    Reinhardt, Martin James; Perry Evenstad, Jan; Faircloth, Susan

    2012-01-01

    Data from this preliminary study, the American Indian--Dads and Daughters Survey, shed light on how American Indian fathers think and feel about their relationships with their daughters. Respondents represent an array of tribal affiliations, age, occupations, socioeconomic status, and geographical/geopolitical locations, helping to ensure that…

  7. Screening of Aspergillus nidulans metabolites from habitat mimicking media using LC-DAD-TOFMS system

    DEFF Research Database (Denmark)

    Klitgaard, Andreas; Holm, Dorte Koefoed; Frisvad, Jens Christian;

    2012-01-01

    cultured on a range of different habitat mimicking media. Extracts from the fungi were analyzed using a LC-DAD-TOFMS system, and by screening the extracts using an in-house database of known fungal metabolites. Using this database we were able to easily select unknown compounds, and dereplicate these using...

  8. Acrylamide in Romanian food using HPLC-UV and a health risk assessment.

    Science.gov (United States)

    Oroian, Mircea; Amariei, Sonia; Gutt, Gheorghe

    2015-01-01

    The aim of this study was to investigate the level of acrylamide from coffee, potato chips and French fries in Romanian food. According to the European Food Safety Authority, coffee beans, potato chips and French fries have the highest levels of acrylamide. For this survey, 50 samples of coffee beans, 50 samples of potato chips and 25 samples of French fries were purchased from different producers from the Romanian market. Acrylamide levels have been quantified using high-performance liquid chromatography with a diode array detector (HPLC-DAD) method, using water as mobile phase. Health risk assessment was achieved by computing the average daily intake, hazard quotient, cumulative risk, carcinogenic risk and cancer risk. For coffee, potato chips and French fries, acrylamide was not shown to pose a health risk in Romanian food.

  9. Development of an in-line HPLC fingerprint ion-trap mass spectrometric method for identification and quality control of Radix Scrophulariae.

    Science.gov (United States)

    Jing, Jing; Chan, Chi-on; Xu, Lijia; Jin, Dengping; Cao, Xinwei; Mok, Daniel K W; Parekh, H S; Chen, Sibao

    2011-12-05

    Chromatographic fingerprinting has been widely accepted as a crucial method for qualitative and quantitative analyses of bioactives within traditional Chinese medicine. A fingerprint provides detailed information, specific for any given herb, thus facilitating the quality control measures of a given traditional Chinese medicine. In this article, quality assessment of Radix Scrophulariae was achieved by using high performance liquid chromatography combining diode-array detection and electrospray ionization mass spectrometry (HPLC-DAD-ESI/MS). Eight batches of sample obtained from different origins in China were used to establish the fingerprint and quantitative analyses. By comparing the retention times, UV and MS spectral data with reference standards, four characteristic peaks in the chromatograms were confirmed as corresponding to acetoside, angoroside C, cinnamic acid, and harpagoside. In addition, other two characteristic peaks were tentatively identified, following the literature interpretation of HPLC-ESI-MS and LC-MS/MS (affording structural information) to be sibirioside A and scrophuloside B(4), respectively. The results indicated that the newly developed HPLC-DAD-MS fingerprint method would be suitable for quality control of Radix Scrophulariae.

  10. Downregulation of LIMK1–ADF/cofilin by DADS inhibits the migration and invasion of colon cancer

    Science.gov (United States)

    Su, Jian; Zhou, Yujuan; Pan, Zhibing; Shi, Ling; Yang, Jing; Liao, Aijun; Liao, Qianjin; Su, Qi

    2017-01-01

    This study aimed to explore whether the downregulation of LIM kinase 1 (LIMK1)-actin depolymerization factor (ADF, also known as destrin)/cofilin by diallyl disulfide (DADS) inhibited the migration and invasion of colon cancer. Previous studies have shown that silencing LIMK1 could significantly enhance the inhibitory effect of DADS on colon cancer cell migration and invasion, suggesting that LIMK1 was a target molecule of DADS, which needed further confirmation. This study reported that LIMK1 and destrin were highly expressed in colon cancer and associated with poor prognosis of patients with colon cancer. Also, the expression of LIMK1 was positively correlated with the expression of destrin. The overexpression of LIMK1 significantly promoted colon cancer cell migration and invasion. DADS obviously inhibited migration and invasion by suppressing the phosphorylation of ADF/cofilin via downregulation of LIMK1 in colon cancer cells. Furthermore, DADS-induced suppression of cell proliferation was enhanced and antagonized by the knockdown and overexpression of LIMK1 in vitro and in vivo, respectively. Similar results were observed for DADS-induced changes in the expression of vimentin, CD34, Ki-67, and E-cadherin in xenografted tumors. These results indicated that LIMK1 was a potential target molecule for the inhibitory effect of DADS on colon cancer cell migration and invasion. PMID:28358024

  11. The Expression of Water and Ion Channels in Diffuse Alveolar Damage Is Not Dependent on DAD Etiology

    Science.gov (United States)

    Del Carlo Bernardi, Fabiola; Alves de Araujo, Priscila; Mauad, Thais; Dolhnikoff, Marisa

    2016-01-01

    Introduction Aquaporins and ion channels are membrane proteins that facilitate the rapid movement of water and solutes across biological membranes. Experimental and in vitro studies reported that the function of these channels and pulmonary edema resolution are impaired in acute lung injury (ALI). Although current evidence indicates that alveolar fluid clearance is impaired in patients with ALI/diffuse alveolar damage (DAD), few human studies have addressed the alterations in pulmonary channels in this clinical condition. Additionally, it is not known whether the primary cause of DAD is a relevant variable for the channel dysfunction. Methods Autopsied lungs of 43 patients with acute respiratory failure (ARF) due to DAD of three different etiologies, non-pulmonary sepsis, H1N1 viral infection and leptospirosis, were compared to 18 normal lungs. We quantified the expression of aquaporin (AQP) 1, AQP3, AQP5, epithelial Na+ channel (ENaC) and sodium potassium ATPase (Na-K-ATPase) in the alveolar septum using immunohistochemistry and image analysis. Results The DAD group presented with increased expression of AQP3, AQP5 and Na-K-ATPase and decreased expression of ENaC compared to controls. However, there was no difference in protein expression within the DAD groups of different etiologies. Conclusion Water and ion channels are altered in patients with ARF due to DAD. The cause of DAD does not seem to influence the level of impairment of these channels. PMID:27835672

  12. A new fluorescent assay for sialyltransferase.

    Science.gov (United States)

    Kajihara, Y; Kamitani, T; Sakakibara, T

    2001-04-23

    A new fluorescent assay for the sialyltransferase reaction was established. After incubation of the sialyltransferase reaction, the sialyloligosaccharide obtained was treated by acid hydrolysis, and then the NeuAc that was released was labeled with 1,2-diamino-4,5-methylenedioxibenzene. The fluorescent-labeled NeuAc could be estimated by HPLC (excitation: 373 nm; emission: 448 nm) and a Lineweaver-Burk plot could be plotted with the data from this analysis.

  13. Bioanalytical studies of porphyric disorders using HPLC with fluorescence detection

    OpenAIRE

    Cypriani,Michele G. S. C.; Alves,Atecla N. L.; Etelvino J. H. Bechara; Assunção,Nilson A.

    2014-01-01

    We describe here the development, validation, quantification and application of a method for determination of heme porphyrin precursors in the urine of porphyric patients. The isomers coproporphyrinogen I and III (COPRO I and III), uroporphyrinogen I (URO I), heptacarboxylporphyrinogen I (HEPTA I), pentacarboxylporphyrinogen (PENTA I), and hexacarboxylporphyrinogen I (HEXA I) were analyzed. These six urinary heme precursors were determined in urine samples collected from 24 patients by high-p...

  14. A NEW REVERSE PHASE HPLC METHOD WITH FLUORESCENT ...

    African Journals Online (AJOL)

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    (Tuttlingen, Germany) was utilized to centrifuge plasma samples. ... was dissolved in 100 mL of double distilled water to produce its stock solution (1 .... By this measure, the peak tailing which results from ion-exchange interactions was easily.

  15. HPLC profiling of Phellinus linteus.

    Science.gov (United States)

    Kojima, Kazuo; Ogihara, Yukio; Sakai, Yoshimichi; Mizukami, Hajime; Nagatsu, Akito

    2008-10-01

    HPLC chromatograms of MeOH extracts from a fruit body of the wild-grown P. linteus (natural fruit body), from cultivated fungus (cultivated fruit body), and from the cultured mycelia were compared. The extract prepared from the natural fruit bodies revealed a typical HPLC profile referred to as type 1 with a major peak corresponding to meshimakobnol A (1) together with two minor peaks of hypholomine B (3) and inoscavin A (4); the cultivated fruit bodies exhibited a profile referred to as type 2 with major peaks corresponding to 3 and 4 and a minor peak of 1, and the cultured mycelia showed a profile referred to as type 3 without any of these peaks. We also analyzed HPLC chromatograms of commercial products of P. linteus obtained in the markets. Most of the products claimed to be natural fruit bodies exhibited type 1 profiles, except for one product having an intermediate HPLC profile between type 1 and type 2. The products claimed to be cultivated fruit bodies and cultured mycelia revealed type 2 and type 3 profiles, respectively. The present results indicate that the HPLC chromatogram of the methanol extract of P. linteus can be used as a fingerprint to identify whether the product is from natural fruit bodies, cultivated fruit bodies, or cultured mycelia.

  16. Determinação de 3,4-metilenodioximetanfetamina (MDMA em comprimidos de Ecstasy por cromatografia líquida de alta eficiência com detecção por fluorescência (CLAE-DF Determination of 3,4-methylenedioxymethamphetamine (MDMA in Ecstasy tablets by high performance liquid chromatography with fluorescence detection (HPLC-FD

    Directory of Open Access Journals (Sweden)

    José Luiz da Costa

    2009-01-01

    Full Text Available This paper describes the development and validation of simple and selective analytical method for determination of 3.4-methylenedioxymethamphetamine (MDMA in Ecstasy tablets, using high performance liquid chromatography with fluorescence detection. Analysis was performed in a reversed phase column (LiChrospher 100 C18, 150 x 4.6 mm, 5 µm, isocratic elution with phosphate buffer 25 mmol/L pH 3.0 and acetonitrile (95:5, v/v. The method presents adequate linearity, selectivity, precision and accuracy. MDMA concentration in analyzed tablets showed a remarkable variability (from 8.5 to 59.5 mg/tablet although the tablet weights were uniform, indicating poor manufacturing control thus imposing additional health risks to the users.

  17. Fluorescence spectroscopy

    DEFF Research Database (Denmark)

    Bagatolli, Luis

    2016-01-01

    Fluorescence spectroscopy is a powerful experimental tool used by scientists from many disciplines. During the last decades there have been important developments on distinct fluorescence methods, particularly those related to the study of biological phenomena. This chapter discusses...

  18. New rapid methods for determination of total LAS in sewage sludge by high performance liquid chromatography (HPLC) and capillary electrophoresis (CE)

    Energy Technology Data Exchange (ETDEWEB)

    Villar, M. [Department of Analytical Chemistry, Faculty of Chemistry, University of Seville, c/Profesor Garcia Gonzalez s/n, 41012 Seville (Spain); Callejon, M. [Department of Analytical Chemistry, Faculty of Chemistry, University of Seville, c/Profesor Garcia Gonzalez s/n, 41012 Seville (Spain)], E-mail: mochon@us.es; Jimenez, J.C.; Alonso, E.; Guiraum, A. [Department of Analytical Chemistry, Faculty of Chemistry, University of Seville, c/Profesor Garcia Gonzalez s/n, 41012 Seville (Spain)

    2009-02-23

    Linear alkylbenzene sulfonates (LAS) are the most common synthetic anionic surfactant used in domestic and industrial detergents, with a global production of 2.4 x 10{sup 6} t year{sup -1}. After use and disposal, LAS may enter the environment by one of the several routes, including by direct discharge to surface water or discharge to water from sewage treatment plants. Sewage treatment plants break down LAS only partly: some of them remain in effluent and other fraction is adsorbed in sewage solid. New and rapid methods for determination of total LAS from sewage sludge based on microwave assisted extraction and HPLC-FL and CE-DAD determination are proposed. The extraction of total LAS is carried out by using microwaves energy, an extraction time of 10 min and 5 mL of methanol. For HPLC-FL determination, mobile phase acetonitrile-water was used, comprising 60% (v/v) from 0 to 1 min and a flow rate of 1 mL min{sup -1} programmed to 100% acetonitrile between 1 and 2 min and a flow rate of 2 mL min{sup -1}. The final composition was maintained for a further 5 min. The determination of total LAS by CE-DAD was performed in a phosphate buffer (10 mM, pH 9). The separation voltage was 25 kV and the temperature of the capillary was 30 deg. C. Injections were performed in the pressure mode and the injection time was set at 12 s. The determination of total LAS is carried out in less than 5 min. The methods did not require clean-up or preconcentration steps. Detection limit for total LAS in the sludge was 3.03 mg kg{sup -1} using HPLC-FL and 21.0 mg kg{sup -1} using CE-DAD, and recoveries were >85% using both determination methods. Concentrations of total LAS obtained using both methods were compared with the sum of concentrations of homologues LAS C-10, LAS C-11, LAS C-12 and LAS C-13 obtained using microwaves assisted extraction and HPLC-FL and CE-DAD determination.

  19. Acute interstitial pneumonia (AIP): relationship to Hamman-Rich syndrome, diffuse alveolar damage (DAD), and acute respiratory distress syndrome (ARDS).

    Science.gov (United States)

    Mukhopadhyay, Sanjay; Parambil, Joseph G

    2012-10-01

    Acute interstitial pneumonia (AIP) is a term used for an idiopathic form of acute lung injury characterized clinically by acute respiratory failure with bilateral lung infiltrates and histologically by diffuse alveolar damage (DAD), a combination of findings previously known as the Hamman-Rich syndrome. This review aims to clarify the diagnostic criteria of AIP, its relationship with DAD and acute respiratory distress syndrome (ARDS), key etiologies that need to be excluded before making the diagnosis, and the salient clinical features. Cases that meet clinical and pathologic criteria for AIP overlap substantially with those that fulfill clinical criteria for ARDS. The main differences between AIP and ARDS are that AIP requires a histologic diagnosis of DAD and exclusion of known etiologies. AIP should also be distinguished from "acute exacerbation of IPF," a condition in which acute lung injury (usually DAD) supervenes on underlying usual interstitial pneumonia (UIP)/idiopathic pulmonary fibrosis (IPF).

  20. PinK: A TCL/TK based Database Interface to ADAMO and DAD

    Science.gov (United States)

    Ackerstaff, K.; Funk, M.-A.; Düren, M.; Ferstl, M.; Wander, W.; Green, P.; Oelwein, Ph.; Welch, P.

    PINK is a TCL/TK extension developed initially for the HERMES experiment at HERA/DESY. Like other HEP experiments, HERMES needs a highly structured database to cope with the complexity and amount of data being handled. HERMES uses the ADAMO Entity Relationship database concept together with the DAD package which extends the use of ADAMO towards a distributed database with flexible, fast I/O and time dependent data handling. With a large and complex time dependent database and many programs accessing it, there is a strong demand for a flexible common user interface. PINK incorporates the functionality of ADAMO and DAD providing an Object Oriented interface to the database. It can be used as an interactive command line, as well as a graphical user interface to access and manipulate data on the database servers and files.

  1. Determination and characterization of phytochelatins by liquid chromatography coupled with on line chemical vapour generation and atomic fluorescence spectrometric detection.

    Science.gov (United States)

    Bramanti, Emilia; Toncelli, Daniel; Morelli, Elisabetta; Lampugnani, Leonardo; Zamboni, Roberto; Miller, Keith E; Zemetra, Joseph; D'Ulivo, Alessandro

    2006-11-10

    Liquid chromatography (LC) coupled on line with UV/visible diode array detector (DAD) and cold vapour generation atomic fluorescence spectrometry (CVGAFS) has been developed for the speciation, determination and characterization of phytochelatins (PCs). The method is based on a bidimensional approach, e.g. on the analysis of synthetic PC solutions (apo-PCs and Cd(2+)-complexed PCs) (i) by size exclusion chromatography coupled to UV diode array detector (SEC-DAD); (ii) by the derivatization of PC -SH groups in SEC fractions by p-hydroxymercurybenzoate (PHMB) and the indirect detection of PC-PHMB complexes by reversed phase liquid chromatography coupled to atomic fluorescence detector (RPLC-CVGAFS). MALDI-TOF/MS (matrix assisted laser desorption ionization time of flight mass spectrometry) analysis of underivatized synthetic PC samples was performed in order have a qualitative information of their composition. Quantitative analysis of synthetic PC solutions has been performed on the basis of peak area of PC-PHMB complexes of the mercury specific chromatogram and calibration curve of standard solution of glutathione (GSH) complexed to PHMB (GS-PHMB). The limit of quantitation (LOQ) in terms of GS-PHMB complex was 90 nM (CV 5%) with an injection volume of 35 microL, corresponding to 3.2 pmol (0.97 ng) of GSH. The method has been applied to analysis of extracts of cell cultures from Phaeodactylum tricornutum grown in Cd-containing nutrient solutions, analysed by SEC-DAD-CVGAFS and RPLC-DAD-CVGAFS.

  2. Determination of lupenone and β-sitosterol in Rhizoma Musae by UPLC with DAD and ELSD.

    Science.gov (United States)

    Xu, Feng; Wu, Hongmei; Wang, Yuanmin; Yang, Ye; Gao, Yuan; Wang, Xiangpei

    2015-01-01

    Rhizoma Musae has been used for centuries in Miao medicine practice in China, and it usually uses for treating diabetes and bruises. In this study, lupenone and β-sitosterol in Rhizoma Musae were separated by reversed-phase ultra-performance liquid chromatography (RP-UPLC) and simultaneously detected by a diode array detector (DAD) and an evaporative light scattering detector (ELSD) using methanol and 0.1% aqueous acetic acid (100 : 4, v/v) as a mobile phase in 20 min. The flow rate of 0.1 mL/min was set with isocratic, the temperature of column compartment maintained at 50°C and ultraviolet detection set at 206 nm wavelength. The injection volume was 1.0 µL. The parameter for the ELSD was set to a probe temperature of 45°C, and the nebulizer for nitrogen gas was adjusted to 1.5 L/min. The RP-UPLC method was validated for accuracy, precision, limit of detection and limit of quantification. It applied to the quantification of the active chemical constituents of Rhizoma Musae, and results indicated that both DAD and ELSD were suitable for the determination of lupenone and β-sitosterol, and the DAD has a better sensitivity than the ELSD.

  3. Rapid resolution liquid chromatography-mass spectrometry and high-performance liquid chromatography-fluorescence detection for metabolism and pharmacokinetic studies of ergosta-4,6,8(14),22-tetraen-3-one.

    Science.gov (United States)

    Zhao, Ying-Yong; Qin, Xiang-Yang; Cheng, Xian-Long; Liu, Xue-Ying; Lin, Rui-Chao; Zhang, Yongmin; Li, Xiao-Ye; Sun, Xiao-Li; Sun, Wen-Ji

    2010-08-24

    Ergosta-4,6,8(14),22-tetraen-3-one (ergone) from many medicinal plants has been demonstrated to possess a variety of pharmacological activities in vivo and in vitro, including cytotoxic, diuretic and immunosuppressive activity. Metabolism and pharmacokinetic studies on rat were conducted for ergone. Rapid resolution liquid chromatography with atmospheric pressure chemical ionization tandem multi-stage mass spectrometry (RRLC-APCI-MS(n)) and high-performance liquid chromatography with fluorescence detection (HPLC-FLD) methods were applied for the identification and quantification of ergone and its metabolite from rat plasma, faeces and urine. A metabolite was identified by RRLC-DAD-APCI-MS(n): 22,23-epoxy-ergosta-4,6,8(14)-triaen-3-one (epoxyergone). The concentrations of the analyte with its metabolites were determined by HPLC-FLD at excitation wavelength of 370 nm and emission wavelength of 485 nm. The samples were deproteinized with methanol after addition of camptothecin as internal standard (IS). The analysis was performed on a Diamonsil C18 column (150 mm x 4.6 mm x 5 microm) with a mobile phase gradient consisting of methanol and water at a flow rate of 1 mL min(-1). The assay was linear over the concentration range of 42-1500, 36-7500 and 42-1500 ng mL(-1) for plasma, faecal homogenate and urine respectively. The absolute recoveries were found to be 97.0+/-1.2%, 98.1+/-0.7% and 96.6+/-1.8% for plasma, faecal homogenate and urine respectively. The intra-day and inter-day relative standard deviations (RSD) were less than 10%. The previous HPLC-MS/MS method is not affordable for most laboratories because of the specialty requirement and high equipment cost. However, the HPLC-FLD method is economic and operating simply for quantitative determination of ergone and its metabolite in rat plasma, faeces and urine. In addition, liquid chromatography coupled with ion trap multi-stage mass spectrometry is becoming a useful technique for ergone metabolite identification.

  4. HPLC analysis of in vivo intestinal absorption and oxidative metabolism of salicylic acid in the rat.

    Science.gov (United States)

    Kuzma, Mónika; Nyúl, Eszter; Mayer, Mátyás; Fischer, Emil; Perjési, Pál

    2016-12-01

    In vivo absorption and oxidative metabolism of salicylic acid in rat small intestine was studied by luminal perfusion experiment. Perfusion through the lumen of proximal jejunum with isotonic medium containing 250 μm sodium salicylate was carried out. Absorption of salicylate was measured by a validated HPLC-DAD method which was evaluated for a number of validation characteristics (specificity, repeatability and intermediate precision, limit of detection, limit of quantification, linearity and accuracy). The method was linear over the concentration range 0.5-50 μg/mL. After liquid-liquid extraction of the perfusion samples oxidative biotransformation of salicylate was also investigated by HPLC-MS. The method was linear over the concentration range 0.25-5.0 μg/mL. Two hydroxylated metabolites of salicylic acid (2,5-dihydroxybenzoic acid and 2,3-dihydroxybenzoic acid) were detected and identified. The mean recovery of extraction was 72.4% for 2,3-DHB, 72.5% for 2,5-DHB and 50.1% for salicylic acid, respectively. The methods were successfully applied to investigate jejunal absorption and oxidative metabolism of sodium salicylate in experimental animals. The methods provide analytical background for further metabolic studies of salycilates under modified physiological conditions.

  5. Extraction and Determination of Cyproheptadine in Human Urine by DLLME-HPLC Method.

    Science.gov (United States)

    Maham, Mehdi; Kiarostami, Vahid; Waqif-Husain, Syed; Abroomand-Azar, Parviz; Tehrani, Mohammad Saber; Khoeini Sharifabadi, Malihe; Afrouzi, Hossein; Shapouri, Mahmoudreza; Karami-Osboo, Rouhollah

    2013-01-01

    Novel dispersive liquid-liquid microextraction (DLLME), coupled with high performance liquid chromatography with photodiode array detection (HPLC-DAD) has been applied for the extraction and determination of cyproheptadine (CPH), an antihistamine, in human urine samples. In this method, 0.6 mL of acetonitrile (disperser solvent) containing 30 μL of carbon tetrachloride (extraction solvent) was rapidly injected by a syringe into 5 mL urine sample. After centrifugation, the sedimented phase containing enriched analyte was dissolved in acetonitrile and an aliquot of this solution injected into the HPLC system for analysis. Development of DLLME procedure includes optimization of some important parameters such as kind and volume of extraction and disperser solvent, pH and salt addition. The proposed method has good linearity in the range of 0.02-4.5 μg mL(-1) and low detection limit (13.1 ng mL(-1)). The repeatability of the method, expressed as relative standard deviation was 4.9% (n = 3). This method has also been applied to the analysis of real urine samples with satisfactory relative recoveries in the range of 91.6-101.0%.

  6. Characterization of Flavonoid Subgroups and Hydroxy Substitution by HPLC-MS/MS

    Directory of Open Access Journals (Sweden)

    Dimitrios Tsimogiannis

    2007-03-01

    Full Text Available HPLC-DAD coupled with mass spectrometry in the positive ionization mode was applied to study the fragmentation of twelve selected flavonoids. Compounds belonging to all the major subgroups found in common plants, i.e. flavonols, flavones, dihydroflavonols, flavanones and flavanols were studied. Compound standards were injected into the spectrometer and produced characteristic mass spectra. The fragmentation of each compound was studied and it was shown that the dehydration andcarbon monoxide losses from the [M+H]+ ion by the members of each subgroup produced specific fragments, thus allowing the characterization of the flavonoid subgroups. Moreover, fragments resulting from fission of the C-rings are specific of each subgroup and revealed the substitution pattern of A- and B-rings. In order to verify the identifying efficiency of the positive ionization mode through these characteristic fragmentations, the unknown flavonoids of an Origanum vulgare diethyl ether extract were separated withthe HPLC system and the major peaks were successfully identified with the mass spectrometer.

  7. SPE and HPLC/UV of resin acids in colophonium-containing products.

    Science.gov (United States)

    Nilsson, Ulrika; Berglund, Naghmeh; Lindahl, Fredrik; Axelsson, Sara; Redeby, Theres; Lassen, Pia; Karlberg, Ann-Therese

    2008-08-01

    A new method, involving SPE and HPLC/UV diode-array detection (DAD), was developed for the quantification of colophonium components in different consumer products, such as cosmetics. Colophonium is a common cause of contact dermatitis since its components can oxidize into allergens on exposure to air. Three different resin acids were used as markers for native and oxidized colophonium, abietic acid (AbA), dehydroabietic acid (DeA), and 7-oxodehydroabietic acid (7-O-DeA). The SPE method, utilizing a mixed-mode hydrophobic and anion exchange retention mechanism, was shown to yield very clean extracts. The use of a urea-embedded C(12) HPLC stationary phase improved the separation of the resin acids compared to common C(18). Concentrations higher than 2 mg/g of both AbA and DeA were detected in wax strips. In this product also 7-O-DeA, a marker for oxidized colophonium, was detected at a level of 28 microg/g. The LODs were in the range of 7-19 microg/g and the LOQs 22-56 microg/g. The method is simple to use and can be applied on many types of technical products, not only cosmetics. For the first time, a method for technical products was developed, which separates AbA from pimaric acid.

  8. Resonance Rayleigh scattering for detection of proteins in HPLC.

    Science.gov (United States)

    Lu, Xin; Luo, Zhihui; Liu, Chengwei; Zhao, Shulin

    2008-09-01

    An HPLC-resonance Rayleigh scattering (RRS) (HPLC-RRS) detection system is described for separation and detection of proteins. This system is based on the modification of a commercial HPLC instrument involving the addition of a pump and a T-shaped interface, and a common fluorescence detector was used for detection. The detection principle is based on the change of RRS intensity of the ion-association complex formed from biebrich scarlet (BS) and protein. The RRS signal was detected at lambdaex=lambdaem=376 nm. The utility of the presented method was demonstrated by the separation and determination of four proteins involving cytochrome (Cyt-c), lysozyme (Lys), HSA, and gamma-globulin (gamma-Glo). An LOD of 0.2-1.0 microg/mL was reached and a linear range was found between peak area and concentration in the range of 0.20-3.0 microg/mL for Cyt-c, 0.25-2.5 microg/mL for Lys, 1.5-10 microg/mL for HSA, and 2.0-15 microg/mL for gamma-Glo, with linear regression coefficients all above 0.99. The method presented has been applied to determine HSA and gamma-Glo in human serum samples synchronously.

  9. The novel technique of vapor pressure analysis to monitor the enzymatic degradation of PHB by HPLC chromatography.

    Science.gov (United States)

    Polyák, Péter; Rácz, Piroska; Rózsa, Péter; Nagy, Gergely N; Vértessy, Beáta G; Pukánszky, Béla

    2017-03-15

    A novel method was introduced for the quantitative determination of substances in aqueous solutions by using the evaporative light scattering (ELS) detector of a high performance liquid chromatograph (HPLC). The principle of the measurement is the different equilibrium vapor pressure of the solvent and the analyte resulting in decreasing evaporation rate, larger droplets and stronger signal with increasing concentration. The new technique based on vapor pressure analysis was validated with traditional UV-Vis detection carried out with a diode array detector (DAD). The new technique was used for monitoring the concentration of solutions obtained during the enzymatic degradation of poly(3-hydroxybutyrate) yielding the 3-hydroxybutyrate monomer as the product. The accuracy of the measurement allowed the determination of degradation kinetics as well. The results obtained with the two techniques showed excellent agreement at small concentrations. Deviations at larger concentrations were explained with the non-linear correlation between analyte concentration and detector signal and the linear regression used for calibration. Mathematical analysis of the method made possible the determination of the evaporation enthalpy of the analyte as well. The new approach is especially suitable for the quantitative analysis of compounds, which do not absorb in the detection range of the DAD detector or if their characteristic absorbance is close to the lower end of its wavelength range.

  10. Rapid screening of transferrin-binders in the flowers of Bauhinia blakeana Dunn by on-line high-performance liquid chromatography-diode-array detector-electrospray ionization-ion-trap-time-of-flight-mass spectrometry-transferrin-fluorescence detection system.

    Science.gov (United States)

    Liu, Meixian; Dong, Jing; Lin, Zongtao; Niu, Yanyan; Zhang, Xiaotian; Jiang, Haixiu; Guo, Ning; Li, Wei; Wang, Hong; Chen, Shizhong

    2016-06-10

    Transferrin (Transferrin, TRF, TF) has drawn increasing attention in cancer therapy due to its potential applications in drug delivery. TF receptor, highly expressed in tumor cells, recognizes and transports Fe(3+)-TF into cells to release iron into cytoplasm. Thus, discovering TF-binding compounds has become an active research area and is of great importance for target therapy. In this study, an on-line analysis method was established for screening TF-binding compounds from the flowers of Bauhinia blakeana Dunn using a high-performance liquid chromatography-diode-array detector-multi-stage mass spectrometry-transferrin-fluorescence detector (HPLC-DAD-MS(n)-TF-FLD) method. As a result, 33 of 80 identified or tentatively characterized compounds in the sample were TF-binding active. Twenty-five flavonol glycosides and eight phenolic acids were identified as TF-binders. Twelve of these active compounds together with six standard compounds were used to study the dose-response effects and structure-activity relationships of flavonoids and phenolic acids. The method was validated by vitexin with a good linearity in the range of concentrations used in the study. The limit of detection for vitexin was 0.1596 nmol. Our study indicated that the established method is simple, rapid and sensitive for screening TF-binding active compounds in the extract of Bauhinia blakeana Dunn, and therefore is important for discovering potential anti-cancer ingredients from complex samples for TF related drug delivery.

  11. Fingerprint analysis and simultaneous determination of phenolic compounds in extracts of Curculiginis Rhizoma by HPLC-diode array detector.

    Science.gov (United States)

    Bian, Qingya; Yang, Hui; Chan, Chi-On; Jin, Dengping; Mok, Daniel Kam-Wah; Chen, Sibao

    2013-01-01

    Curculiginis Rhizoma (Curculigo orchioides GAERTN.) is a well-known Chinese herbal medicine, as well as an important Rasayana drug in India. Current criteria of quality control on this herb are to quantitatively analyze single compound curculigoside, which fail to comprehensively evaluate quality of this herb. In this paper, a simple and reliable HPLC coupled with diode array detector (DAD) method was developed to evaluate the quality of Curculiginis Rhizoma through establishing chromatographic fingerprint and simultaneously quantitating four phenolic compounds, orcinol glucoside, orcinol, 2,6-dimethoxybenzoic acid and curculigoside. The fingerprint displayed eleven common peaks, and the similarity index of different samples was in a range of 0.890-0.977. Validation of the method was acceptable, with 96.03-102.82% accuracy in recovery test and inter and intra-day precisions were less than 2%. This developed method by having a combination of chromatographic fingerprint and quantitation analysis could be applied to the quality control of Curculiginis Rhizoma.

  12. Profiles of urine samples taken from Ecstasy users at Rave parties: analysis by immunoassays, HPLC, and GC-MS.

    Science.gov (United States)

    Zhao, H; Brenneisen, R; Scholer, A; McNally, A J; ElSohly, M A; Murphy, T P; Salamone, S J

    2001-01-01

    The abuse of the designer amphetamines such as 3,4-methylenedioxymethamphetamine (MDMA, Ecstasy) is increasing throughout the world. They have become popular drugs, especially at all-night techno dance parties (Raves), and their detection is becoming an important issue. Presently, there are no MDMA- or MDA-specific immunoassays on the market, and detection of the designer amphetamines is dependent upon the use of commercially available amphetamine assays. The success of this approach has been difficult to assess because of the general unavailability of significant numbers of samples from known drug users. The objectives of the present study are to characterize the drug content of urine samples from admitted Ecstasy users by chromatographic methods and to assess the ability of the available amphetamine/methamphetamine immunoassays to detect methylenedioxyamphetamines. We found that, when analyzed by high-performance liquid chromatography with diode-array detection (HPLC-DAD), 64% of 70 urine samples (by gas chromatography-mass spectrometry [GC-MS]: 88% of 64 urine samples) obtained from Rave attendees contained MDMA and/or 3,4-methylenedioxyamphetamine (MDA) alone or in combination with amphetamine, methamphetamine, or other designer amphetamines such as 3,4-methylenedioxyethylamphetamine (MDEA). This suggests that the majority of the Ravers are multidrug users. At the manufacturer's suggested cutoffs, the Abbott TDx Amphetamine/Methamphetamine II and the new Roche HS Amphetamine/MDMA assays demonstrated greater detection sensitivity for MDMA than the other amphetamine immunoassays tested (Abuscreen OnLine Hitachi AMPS, Abuscreen OnLine Integra AMPS, Abuscreen OnLine Integra AMPSX, CEDIA AMPS, and EMIT II AMPS). There is 100% agreement between each of the two immunoassays with the reference chromatographic methods, HPLC-DAD and GC-MS, for the detection of methylenedioxyamphetamines.

  13. Analysis of xanthines in beverages using a fully automated SPE-SPC-DAD hyphenated system

    Energy Technology Data Exchange (ETDEWEB)

    Medvedovici, A. [Bucarest Univ., Bucarest (Romania). Faculty of Chemistry, Dept. of Analytical Chemistry; David, F.; David, V.; Sandra, P. [Research Institute of Chromatography, Kortrijk (Belgium)

    2000-08-01

    Analysis of some xanthines (caffeine, theophylline and theobromine) in beverages has been achieved by a fully automated on-line Solid Phase Extraction - Supercritical Fluid Chromatography - Diode Array Detection (Spe - Sofc - Dad). Three adsorbents have been tested for the Spe procedure: octadecyl modified silicagel (ODS) and two types of styrene-divinylbenzen copolymer based materials, from which Porapack proved to be the most suitable adsorbent. Optimisation and correlation of both Spe and Sofc operational parameters are also discussed. By this technique, caffeine was determined in ice tea and Coca-Cola in a concentration of 0.15 ppm, theobromine - 1.5 ppb, and theophylline - 0.15 ppb. [Italian] Si e' realizzata l'analis di alcune xantine (caffeina, teofillina e teobromina) mediante un sistema, in linea, completamente automatizzato basato su Estrazione in Fase Solida - Cromatografia in Fase Supercritica - Rivelazione con Diode Array (Spe - Sfc - Dad). Per la procedura Spe sono stati valutati tre substrati: silice ottadecilica (ODS) e due tipi di materiali polimerici a base stirene-divinilbenzene, di cui, quello denominato PRP-1, e' risultato essere il piu' efficiente. Sono discusse sia l'ottimizzazione che la correlazione dei parametri operazionali per la Spe e la Sfc. Con questa tecnica sono state determinate, in te' ghiacciato e Coca-Cola, la caffeina, la teobromina e la teofillina alle concentrazini di 0.15, 1.5 e 0.15 ppm.

  14. LC-DAD-MS (ESI+) analysis and antioxidant capacity of crocus sativus petal extracts.

    Science.gov (United States)

    Termentzi, Aikaterini; Kokkalou, Eugene

    2008-04-01

    In this study, various fractions isolated from the petals of Crocus sativus were assessed at first for their phenolic content both qualitatively and quantitatively and secondly for their antioxidant activity. The phytochemical analysis was carried out by LC-DAD-MS (ESI (+)) whereas the antioxidant potential was evaluated by applying two methodologies, the DPPH. radical scavenging activity test and the Co(II)-induced luminol chemiluminescence procedure. According to data obtained from these antioxidant tests, the diethyl ether, ethyl acetate and aqueous fractions demonstrated the strongest antioxidant capacity. Interestingly, the major constituents identified in these fractions correspond to kaempferol, quercetin, naringenin and some flavanone and flavanol derivatives glycosylated and esterified with phenylpropanoic acids. In addition, the presence of some nitrogen-containing substances, as well as other phenolics and phenylpropanoic derivatives was also traced. The identification and structural elucidation of all substances isolated in this study was achieved by both comparing available literature data and by proposed fragmentation mechanisms based on evaluating the LC-DAD-MS (ESI (+)) experimental data. The quantitative analysis data obtained thus far have shown that Crocus sativus petals are a rich source of flavonoids. Such a fact suggests that the good antioxidant capacity detected in the various fractions of Crocus sativus petals could be attributed to the presence of flavonoids, since it is already known that these molecules exert antioxidant capability. The latter, along with the use of Crocus sativus in food and pharmaceutical industry is discussed.

  15. Structure dependent antioxidant capacity of phlorotannins from Icelandic Fucus vesiculosus by UHPLC-DAD-ECD-QTOFMS.

    Science.gov (United States)

    Hermund, Ditte B; Plaza, Merichel; Turner, Charlotta; Jónsdóttir, Rosa; Kristinsson, Hordur G; Jacobsen, Charlotte; Nielsen, Kristian Fog

    2018-02-01

    Brown algae are rich in polyphenolic compounds, phlorotannins, which have been found to possess high in vitro antioxidant capacity, especially DPPH radical scavenging activity, due to the high number of hydroxyl groups. Whereas, the overall antioxidant capacity of brown algae extracts has been widely studied, the antioxidant capacity of individual phlorotannins has been rarely explored. The aim of this study was to determine the structure dependant antioxidant capacity of phlorotannins from Icelandic brown algae, Fucus vesiculosus. The antioxidant capacity of individual phlorotannins was determined by an on-line method using liquid chromatography and an electrochemical detector followed by quadrupole Time of Flight mass spectrometry (UHPLC-DAD-ECD-QTOFMS). Tentative structural elucidation of 13 phlorotannin isomers from EAF was obtained by LC-DAD-QTOFMS, ranging from 374 to 870Da. On-line determination of antioxidant capacity of the individual phlorotannins generally showed that low molecular phlorotannins exhibited higher antioxidant capacity and that the capacity decreased with polymerisation. Copyright © 2017 Elsevier Ltd. All rights reserved.

  16. HPLC determination of aflatoxin residues in traditional Chinese medicine Yinpian with post column photochemical derivation and fluorescence detection%HPLC柱后光衍生荧光法测定中药饮片中黄曲霉毒素残留量

    Institute of Scientific and Technical Information of China (English)

    郝爱鱼; 赵丽元; 刘英慧; 王戈; 金红宇; 毕秀玲; 门启鸣

    2012-01-01

    目的:采用免疫亲和净化HPLC柱后紫外光化学衍生荧光检测法研究中药饮片中黄曲霉毒素测定的加样回收率,考察该方法在中药饮片黄曲霉毒素残留测定中的可行性,评价120批中药饮片的污染状况,总结污染规律.方法:样品经溶剂提取、免疫亲和柱净化后由HPLC柱后光衍生荧光检测法测定其中黄曲霉毒素残留量,对每种饮片进行4μg·kg-1(以黄曲霉毒素B1计)的加样回收率考察.结果:黄曲霉毒素B1、B2、G1、G2的最低检测限分别为0.44,0.12,0.41,0.16 pg;线性范围分别为1.74 ~218.0,0.50 ~62.17,1.63 ~203.1,0.62 ~61.94 pg(r>0.9999).所考察饮片中黄曲霉毒素B1、B2、G1、G2总回收率和B1回收率均在60% ~ 120%之间的占87%;被污染的批次占9.2%,其中污染程度超过目前中国药典限度的占6.7%.结论:免疫亲和净化HPLC柱后光衍生荧光检测法具有方法简便,衍生反应稳定,灵敏度高,重现性好,反应系统污染少的优点,适合于大多数中药饮片的黄曲霉毒素检测;种子类被污染的程度严重,应扩大该类中药的监测范围.%Objective: To investigate the recoveries of aflatoxin (AF) residues analysis of traditional Chinese medicine (TCM) Yinpian using immunoaffinity column HPLC method with post column photochemical derivatization and fluorescence detection, and evaluate the feasibility of the method. To evaluate the contamination status of 120 batches of TCM Yinpian and summarize the rule of contamination. Methods: Samples were prepared through organic solvent extraction and cleaned up with immunoaffinity column sequentially; the recoveries of aflatoxins were then determined by adding aflatoxins standard mixture at the level of 4 μg · kg-1 according to aflatoxin B1 content. Finally, aflatoxins B1 ,B2 ,G1 and G2 residue amount in TCM Yinpian was determined. Results:The limit of detection of aflatoxins B1, B2 , G1 ,G2 was 0. 44,0. 12,0. 41,0. 16 pg

  17. Acquisition of HPLC-Mass Spectrometer

    Science.gov (United States)

    2015-08-18

    31-Jan-2015 Approved for Public Release; Distribution Unlimited Final Report: Acquisition of HPLC -Mass Spectrometer The views, opinions and/or findings...published in peer-reviewed journals: Final Report: Acquisition of HPLC -Mass Spectrometer Report Title The acquisition of the mass spectrometer has been a

  18. HPLC chromatofocusing of human immunoglobulins.

    Science.gov (United States)

    Waldrep, J C; Schulte, J R

    1989-03-31

    A method is described for fractionation and analysis of IgA, IgM, and IgG and antibodies in human serum and/or plasma using a combination of HPLC chromatofocusing and immunoassay. A pH 9.0-3.2 gradient is utilized to separate the major proteins in the complex biological samples and monoclonal antibody based ELISAs used to determine the isotype profiles. Antigen-specific ELISAs are subsequently utilized to determine the distribution of antibody species within the chromatofocused specimens. This method is versatile since multiple simultaneous assays can easily be run on each fraction generating extensive qualitative information regarding immunoglobulin classes, subclasses, and antibodies and their distribution profiles. Such spectra will prove useful for experimental kinetic analysis of the humoral immune status of humans and experimental animals.

  19. Identification and Determination of Flavonoids in Astragali Radix by High Performance Liquid Chromatography Coupled with DAD and ESI-MS Detection

    Directory of Open Access Journals (Sweden)

    Yu-Ling Wang

    2011-03-01

    Full Text Available A method for the analysis of flavonoids in Astragali Radix by high-performance liquid chromatography (HPLC combined with photodiode-array detection (DAD and an electrospray ionization (ESI - mass spectrometry (MS was developed. After the samples were extracted with ethanol, the optimum separation conditions for these analytes were achieved using a gradient elution system and a 2.0 × 150 mm Shimadzu VP-ODS column. Eight flavonoids were identified to exist in Astragali Radix based on their characteristic UV data and mass spectra. The concentrations of three major components in this herb—ononin, calycosin and formononetin—were determined by LC/ESI-MS in positive selective ion monitoring (SIM mode. The calibration curves were linear in the range of 0.9~180.0 μg·mL−1 for ononin, 1.8~360.0 μg·mL−1 for calycosin and 1.4~280 μg·mL−1 for formononetin, respectively. The limits of quantification (LOQ and detection (LOD were 0.9 μg· mL−1 and 0.2 μg mL−1 for ononin, 1.8 μg mL−1 and 0.5 μg·mL−1 for calycosin, 1.4 μg mL−1 and 0.5 μg·mL−1 for formononetin, respectively. The standard recoveries were between 95.4~104.7%. The developed method was proven to be useful for the quantitative and qualitative analysis of flavonoid constituents in various resources of Astragali Radix.

  20. Use of multiresponse statistical techniques to optimize the separation of diosmin, hesperidin, diosmetin and hesperitin in different pharmaceutical preparations by high performance liquid chromatography with UV-DAD.

    Science.gov (United States)

    Sammani, Mohamad Subhi; Clavijo, Sabrina; Portugal, Lindomar; Suárez, Ruth; Seddik, Hassan; Cerdà, Víctor

    2017-05-15

    A new method for the separation and determination of four flavonoids: hesperidin (HES), diosmin (DIO), hesperitin (HTIN), and diosmetin (DTIN) in pure form and pharmaceutical formulations has been developed by using high performance liquid chromatography (HPLC) with UV-DAD detection. Multivariate statistics (2(k) full factorial and Box Behnken Designs) has been used for the multiresponse optimization of the chromatographic separation, which was completed in 22min, and carried out on a symmetry® C18 column (250×3mm; 5µm) as stationary phase. Separation was conducted by gradient elution mode using a mixture of methanol, acetonitrile and water pH: 2.5 (CH3COOH), as mobile phase. Analytes were separated setting the column at 22°C, with a flow rate of 0.58mLmin(-1) and detected at 285nm. Under the optimized conditions, the flavonoids showed retention times of: 8.62, 11.53, 18.55 and 19.94min for HES, DIO, HTIN and DTIN, respectively. Limits of detection and quantification were <0.0156µgmL(-1) and <0.100µgmL(-1), respectively. Linearity was achieved with good correlation coefficients values (r(2)=0.999; n=5). Intra-day and inter-day precision were found to be less than 3.44% (n=7). Finally, the proposed method was successfully applied to determine the target flavonoids in pharmaceutical preparations with satisfactory recoveries (between 95.2% and 107.9%), demonstrating that should also find application in the quality control, as well as in the pharmacokinetic studies of these drugs.

  1. Exhaustive Qualitative LC-DAD-MS(n) Analysis of Arabica Green Coffee Beans: Cinnamoyl-glycosides and Cinnamoylshikimic Acids as New Polyphenols in Green Coffee.

    Science.gov (United States)

    Baeza, Gema; Sarriá, Beatriz; Bravo, Laura; Mateos, Raquel

    2016-12-28

    Coffee is one of the most consumed beverages in the world, due to its unique aroma and stimulant properties. Although its health effects are controversial, moderate intake seems to be beneficial. The present work deals with the characterization and quantification of polyphenols and methylxanthines in four Arabica green coffee beans from different geographical origins. The antioxidant activity was also evaluated. Forty-three polyphenols (cinnamic acid, cinnamoyl-amide, 5 cinammoyl-glycosides, and 36 cinnamate esters) were identified using LC-MS(n). Among these, cinnamate esters of six different chemical groups (including two dimethoxycinnamoylquinic acid isomers, three caffeoyl-feruloylquinic acid isomers, caffeoyl-sinapoylquinic acid, p-coumaroyl-feruloylquinic acid, two caffeoylshikimic acid isomers, and trimethoxycinnamoylshikimic acid) in addition to five isomers of cinnamoyl-glycosides called caffeoyl-2,7-anhydro-3-deoxy-2-octulopyranosic acid (CDOA) are described for the first time in Arabica green coffee beans. Moreover, 38 polyphenols (6-7% w/w) and 2 methylxanthines (1.3% w/w) were quantified by HPLC-DAD. Caffeoylquinic was the most abundant group of compounds (up to 85.5%) followed by dicaffeoylquinic and feruloylquinic acids (up to 8 and 7%, respectively) and the newly identified cinnamoyl-glycosides (CDOA) (up to 2.5%). Caffeine was the main methylxanthine (99.8%), with minimal amounts of theobromine (0.2%). African coffees (from Kenya and Ethiopia) showed higher polyphenolic content than American beans (from Brazil and Colombia), whereas methylxanthine contents varied randomly. Both phenols and methylxanthines contributed to the antioxidant capacity associated with green coffee, with a higher contribution of polyphenols. We conclude that green coffee represents an important source of polyphenols and methylxanthines, with high antioxidant capacity.

  2. But ... Dad

    Institute of Scientific and Technical Information of China (English)

    黄蓓

    2005-01-01

    @@ A young boy had just gotten his driving permit. He asked his father, who was a minister, if they could discuss his use of the family car. His father said to him, "I'll make a deal with you. You bring your grades up, study your bible a little, and get your hair cut, and then we will talk about it."

  3. Fluorescent microspheres

    Science.gov (United States)

    Rembaum, A.

    1978-01-01

    Latex particles with attached antibodies have potential biochemical and environmental applications. Human red blood cells and lymphocytes have been labeled with fluorescent microspheres by either direct or indirect immunological technique. Immunolatex spheres can also be used for detecting and localizing specific cell surface receptors. Hormones and toxins may also be bondable.

  4. Analysis of alkylresorcinols in wheat germ oil and barley germ oil via HPLC and flourescence detection: Cochromatography with tocols

    Science.gov (United States)

    Alkylresorcinols are long chain phenolic compounds that have been reported to be localized in the outer layers of the kernels of wheat, rye, barley and other grains. A sensitive HPLC method with fluorescence detection was recently reported for the quantitative analysis of alkylresorcinols in cereal...

  5. Simultaneous Determination of Structurally Diverse Compounds in Different Fangchi Species by UHPLC-DAD and UHPLC-ESI-MS/MS

    Directory of Open Access Journals (Sweden)

    Kang Ro Lee

    2013-05-01

    Full Text Available Two bisbenzylisoquinoline alkaloids, two morphine alkaloids, one aporphine alkaloid, syringaresinol and aristolochic acid І were selected as marker compounds and simultaneously analyzed using an ultra-high pressure liquid chromatography-diode array detection (UHPLC-DAD method. These marker compounds were used for the quality control of Fangchi species of different origins, including Sinomenium acutum, Stephania tetrandra, Cocculus trilobus and Aristolochia fangchi. A reversed-phase UHPLC-DAD method was developed and validated for the simultaneous quantification of structurally diverse markers in different Fangchi species. In addition, an UHPLC-electrospray ionization tandem mass spectrometry (ESI-MS/MS method was used for marker identification in Fangchi species, which provided diagnostic MS/MS spectral patterns that were dependent upon the marker structures. The UHPLC-MS/MS data were used to confirm and complement the UHPLC-DAD quality evaluation results. Additionally, magnoflorine and syringaresinol were observed for the first time in S. tetrandra and C. trilobus, respectively. Twenty different Fangchi species samples were analyzed for aristolochic acid I, syringaresinol and the alkaloids using the UHPLC-DAD and MS/MS method. Based on the levels of markers and principal component analysis (PCA, this method allowed for the clear classification of the samples into four different groups representing samples originating from the four species.

  6. Simultaneous determination of structurally diverse compounds in different Fangchi species by UHPLC-DAD and UHPLC-ESI-MS/MS.

    Science.gov (United States)

    Sim, Hee-Jung; Kim, Ji Hee; Lee, Kang Ro; Hong, Jongki

    2013-05-07

    Two bisbenzylisoquinoline alkaloids, two morphine alkaloids, one aporphine alkaloid, syringaresinol and aristolochic acid І were selected as marker compounds and simultaneously analyzed using an ultra-high pressure liquid chromatography-diode array detection (UHPLC-DAD) method. These marker compounds were used for the quality control of Fangchi species of different origins, including Sinomenium acutum, Stephania tetrandra, Cocculus trilobus and Aristolochia fangchi. A reversed-phase UHPLC-DAD method was developed and validated for the simultaneous quantification of structurally diverse markers in different Fangchi species. In addition, an UHPLC-electrospray ionization tandem mass spectrometry (ESI-MS/MS) method was used for marker identification in Fangchi species, which provided diagnostic MS/MS spectral patterns that were dependent upon the marker structures. The UHPLC-MS/MS data were used to confirm and complement the UHPLC-DAD quality evaluation results. Additionally, magnoflorine and syringaresinol were observed for the first time in S. tetrandra and C. trilobus, respectively. Twenty different Fangchi species samples were analyzed for aristolochic acid I, syringaresinol and the alkaloids using the UHPLC-DAD and MS/MS method. Based on the levels of markers and principal component analysis (PCA), this method allowed for the clear classification of the samples into four different groups representing samples originating from the four species.

  7. Detection of citrinin in ochratoxin A-containing products by a new HPLC method.

    Science.gov (United States)

    Meister, U

    2003-03-01

    A new method for citrinin was developed and validated, which is based on solid phase extraction with polyamide columns and HPLC with fluorescence detection. Sufficient skill with the method given, precise results, i.e. variation coefficients cocoa shells and raisins. Citrinin was detected in 14 OTA-containing samples (1-8 µg/kg). Furthermore, it was demonstrated that citrinin also can be determined in red mold rice according to the new method.

  8. The role of reactive oxygen species (ROS) production on diallyl disulfide (DADS) induced apoptosis and cell cycle arrest in human A549 lung carcinoma cells

    Energy Technology Data Exchange (ETDEWEB)

    Wu Xinjiang [Institute of Indoor and Environmental Toxicology, Faculty of Medicine, Justus-Liebig-University of Giessen, Aulweg 123, D-35385 Giessen (Germany); Kassie, Fekadu [Institute of Indoor and Environmental Toxicology, Faculty of Medicine, Justus-Liebig-University of Giessen, Aulweg 123, D-35385 Giessen (Germany); Mersch-Sundermann, Volker [Institute of Indoor and Environmental Toxicology, Faculty of Medicine, Justus-Liebig-University of Giessen, Aulweg 123, D-35385 Giessen (Germany)]. E-mail: Volker.mersch-sundermann@uniklinikum-giessen.de

    2005-11-11

    Diallyl disulfide (DADS), an oil soluble constituent of garlic (Allium sativum), has been reported to cause antimutagentic and anticarcinogenic effects in vitro and in vivo by modulating phases I and II enzyme activities. In recent years, several studies suggested that the chemopreventive effects of DADS can also be attributed to induction of cell cycle arrest and apoptosis in cancer cells. In the present study, we reported that DADS-induced cell cycle arrest at G2/M and apoptosis in human A549 lung cancer cells in a time- and dose-dependent manner. Additionally, a significant increase of intracellular reactive oxygen species (ROS) was induced in A549 cells less than 0.5 h after DADS treatment, indicating that ROS may be an early event in DADS-modulated apoptosis. Treatment of A549 cells with N-acetyl cysteine (NAC) completely abrogated DADS-induced cell cycle arrest and apoptosis. The result indicated that oxidative stress modulates cell proliferation and cell death induced by DADS.

  9. Nonlinear fluorescence probe using photoinduced charge separation (Presentation Recording)

    Science.gov (United States)

    Mochizuki, Kentaro; Shi, Lanting; Mizukami, Shin; Yamanaka, Masahito; Tanabe, Mamoru; Gong, Wei-Tao; Palonpon, Almar F.; Kawano, Shogo; Kawata, Satoshi; Kikuchi, Kazuya; Fujita, Katsumasa

    2015-08-01

    Two-photon excitation microscopy (TPEM) provides spatial resolution beyond the optical diffraction limit using the nonlinear response of fluorescent molecules. One of the strong advantages of TPEM is that it can be performed using a laser-scanning microscope without a complicated excitation method or computational post-processing. However, TPEM has not been recognized as a super-resolution microscopy due to the use of near-infrared light as excitation source, which provides lower resolution than visible light. In our research, we aimed for the realization of nonlinear fluorescence response with visible light excitation to perform super-resolution imaging using a laser-scanning microscope. The nonlinear fluorescence response with visible light excitation is achieved by developing a probe which provides stepwise two-photon excitation through photoinduced charge separation. The probe named nitro-bisBODIPY consists of two fluorescent molecules (electron donor: D) and one electron acceptor (A), resulting to the structure of D-A-D. Excited by an incident photon, nitro-bisBODIPY generates a charge-separated pair between one of the fluorescent molecules and the acceptor. Fluorescence emission is obtained only when one more incident photon is used to excite the other fluorescent molecule of the probe in the charge-separated state. This stepwise two-photon excitation by nitro-bisBODIPY was confirmed by detection of the 2nd order nonlinear fluorescence response using a confocal microscope with 488 nm CW excitation. The physical model of the stepwise two-photon excitation was investigated by building the energy diagram of nitro-bisBODIPY. Finally, we obtained the improvement of spatial resolution in fluorescence imaging of HeLa cells using nitro-bisBODIPY.

  10. HPLC for quality control of polyimides

    Science.gov (United States)

    Young, P. R.; Sykes, G. F.

    1979-01-01

    High Pressure Liquid Chromatography (HPLC) as a quality control tool for polyimide resins and prepregs are presented. A data base to help establish accept/reject criteria for these materials was developed. This work is intended to supplement, not replace, standard quality control tests normally conducted on incoming resins and prepregs. To help achieve these objectives, the HPLC separation of LARC-160 polyimide precursor resin was characterized. Room temperature resin aging effects were studied. Graphite reinforced composites made from fresh and aged resin were fabricated and tested to determine if changes observed by HPLC were significant.

  11. Expermental Studies of quantitative evaluation using HPLC

    Directory of Open Access Journals (Sweden)

    Ki Rok Kwon

    2005-06-01

    Full Text Available Methods : This study was conducted to carry out quantitative evaluation using HPLC Content analysis was done using HPLC Results : According to HPLC analysis, each BVA-1 contained approximately 0.36㎍ melittin, and BVA-2 contained approximately 0.54㎍ melittin. But the volume of coating was so minute, slight difference exists between each needle. Conclusion : Above results indicate that the bee venom acupuncture can complement shortcomings of syringe usage as a part of Oriental medicine treatment, but extensive researches should be done for further verification.

  12. HPLC for quality control of polyimides

    Science.gov (United States)

    Young, P. R.; Sykes, G. F.

    1979-01-01

    High Pressure Liquid Chromatography (HPLC) as a quality control tool for polyimide resins and prepregs are presented. A data base to help establish accept/reject criteria for these materials was developed. This work is intended to supplement, not replace, standard quality control tests normally conducted on incoming resins and prepregs. To help achieve these objectives, the HPLC separation of LARC-160 polyimide precursor resin was characterized. Room temperature resin aging effects were studied. Graphite reinforced composites made from fresh and aged resin were fabricated and tested to determine if changes observed by HPLC were significant.

  13. Isolation of antioxidative secoiridoids from olive wood (Olea europaea L.) guided by on-line HPLC-DAD-radical scavenging detection

    NARCIS (Netherlands)

    Pérez-Bonilla, M.; Salido, S.; Beek, van T.A.; Waard, de P.; Linares-Palomino, P.J.; Sánchez, A.; Altarejos, J.

    2011-01-01

    The woody portion of olive tree pruning is a source of natural antioxidants of potential interest for the food industry. This work deals with the isolation and identification of further antioxidants present in an ethyl acetate extract of olive (Olea europaea L.) wood. Thus, a new secoiridoid, oleuro

  14. Antioxidant capacity and HPLC-DAD-MS profiling of Chilean peumo (Cryptocarya alba) fruits and comparison with German peumo (Crataegus monogyna) from southern Chile.

    Science.gov (United States)

    Simirgiotis, Mario J

    2013-02-05

    Liquid chromatography (LC) coupled with UV detection and electrospray ionization (ESI) tandem mass spectrometry (MS/MS) was used for the generation of chemical fingerprints and the identification of phenolic compounds in peumo fruits and aerial parts from southern Chile. Thirty three compounds (19 of these detected in C. alba and 23 in C. monogyna) were identified, mainly flavonoid glycosides, phenolic acids, anthocyanins and flavonoid aglycons. Total phenolic content and total flavonoid content was measured for both species, and were higher in the extracts from C. monogyna fruits and aerial parts than extracts from C. alba. The fruits of Cryptocarya alba (Chilean peumo) presented high antioxidant capacity (9.12 ± 0.01 mg/mL in the DPPH assay), but was three times lower to that of Crataegus monogyna (German peumo) (3.61 ± 0.01 mg/mL in the DPPH assay).

  15. Antioxidant Capacity and HPLC-DAD-MS Profiling of Chilean Peumo (Cryptocarya alba Fruits and Comparison with German Peumo (Crataegus monogyna from Southern Chile

    Directory of Open Access Journals (Sweden)

    Mario J. Simirgiotis

    2013-02-01

    Full Text Available Liquid chromatography (LC coupled with UV detection and electrospray ionization (ESI tandem mass spectrometry (MS/MS was used for the generation of chemical fingerprints and the identification of phenolic compounds in peumo fruits and aerial parts from southern Chile. Thirty three compounds (19 of these detected in C. alba and 23 in C. monogyna were identified, mainly flavonoid glycosides, phenolic acids, anthocyanins and flavonoid aglycons. Total phenolic content and total flavonoid content was measured for both species, and were higher in the extracts from C. monogyna fruits and aerial parts than extracts from C. alba. The fruits of Cryptocarya alba (Chilean peumo presented high antioxidant capacity (9.12 ± 0.01 mg/mL in the DPPH assay, but was three times lower to that of Crataegus monogyna (German peumo (3.61 ± 0.01 mg/mL in the DPPH assay.

  16. Development of a targeted GC/MS screening method and validation of an HPLC/DAD quantification method for piperazines–amphetamines mixtures in seized material

    Directory of Open Access Journals (Sweden)

    Yacine Boumrah

    2014-09-01

    Full Text Available Piperazine-related drugs are sold as party pills in the form of tablets, capsules, liquids or powders. These party pills can contain several piperazine derivatives, or even a mixture of piperazines and amphetamine derivatives. This paper describes a screening method using a gas chromatography–mass spectrometry technique allowing the separation and the identification of active components within these mixtures by a combined silylation and acylation derivatization procedure. The studied substances–namely: 1-benzylpiperazine (BZP, 1-(3,4-methylenedioxyben-zylpiperazine (MDBP, 1-(3-trifluoromethylphenylpiperazine (TFMPP, 1-(3-chlorophenyl piperazine (mCPP, 1-(4-methoxyphenyl piperazine (MeOPP, amphetamine, methamphetamine, ephedrine, pseudoephedrine, 3,4-methylenedioxy-N-methamphetamine (MDMA, 3,4-methylenedi-oxyamphetamine (MDA, 3,4-methylenedioxy-N-ethylamphetamine (MDEA and N-methyl-1,3-benzodioxolylbutanamine (MBDB–are separated.

  17. Isolation of antioxidative secoiridoids from olive wood (Olea europaea L.) guided by on-line HPLC-DAD-radical scavenging detection

    NARCIS (Netherlands)

    Pérez-Bonilla, M.; Salido, S.; Beek, van T.A.; Waard, de P.; Linares-Palomino, P.J.; Sánchez, A.; Altarejos, J.

    2011-01-01

    The woody portion of olive tree pruning is a source of natural antioxidants of potential interest for the food industry. This work deals with the isolation and identification of further antioxidants present in an ethyl acetate extract of olive (Olea europaea L.) wood. Thus, a new secoiridoid, oleuro

  18. STRATEGIA ANALITICA PER LA CARATTERIZZAZIONE DEL PROFILO VITAMINICO LIPOSOLUBILE E CAROTENOIDEO DEL LATTE DI DIFFERENTI SPECIE ANIMALI MEDIANTE IFENAZIONE HPLC-DAD-TANDEM MS

    OpenAIRE

    BELLANTE, SIMONA

    2013-01-01

    Vitamin determination is a complex and challenging task, especially when aimed at the analysis of biological and food samples. Milk is a unique matrix, being at the same time biological fluid and food with the chemical characteristics of three phases: emulsion, suspension and colloidal solution. Furthermore, it is an almost complete food and an excellent source of vitamins, in particular vitamins A, B1, B2, B5 and B12. The water-soluble vitamins occur in the serum, while the fat-soluble vitam...

  19. Toxicological screening of human plasma by on-line SPE-HPLC-DAD: identification and quantification of basic drugs and metabolites.

    Science.gov (United States)

    Mut, Ludmila; Grobosch, Thomas; Binscheck-Domaß, Torsten; Frenzel, Wolfgang

    2015-06-01

    An automated multi-analyte screening method for the identification and quantification of 92 drugs and metabolites based on on-line solid-phase extraction-high-performance liquid chromatography-diode array detection technique was developed and successfully validated. In addition, a database with 870 entries including UV-spectra, relative/retention times and response factors of toxicologically relevant compounds was created. Plasma samples (0.2 mL) were treated with methanol, diluted with buffer and on-line extracted (Strata X, 20 ×2 mm, 25 µm) at pH 9. Analytical separation was carried out on a Gemini NX column (150 ×4.6 mm, 3 µm) using gradient elution with acetonitrile-water (90:10,v/v) and 0.05 m potassium dihydrogen phosphate buffer (pH 2.3). Linear calibration curves with correlation coefficients ≥0.9950 were obtained for 78 analytes. As an additional benefit, the newly developed method allows the quantification of 42 analytes (e.g. antidepressants, neuroleptics and anticonvulsants) in a concentration range suitable for therapeutic drug monitoring. Limits of quantitation ranged from 0.02 mg/L (chlordiazepoxide) to 3.4 mg/L (mexiletine). Inter- and intra-day precisions of quality control samples (low/high) were better than 15% (zolpidem) and accuracy (bias) ranged from -11% (opipramol, venlafaxine) to 11% (venlafaxine, trazodone). Tests for carry-over and sample stability under different storage conditions were also performed and stability was adequate. Four cases of poisoning analysis are presented.

  20. Characterisation of an Aromatic Plant-based Formula using UV-Vis Spectroscopy, LC–ESI(+QTOF-MS and HPLC-DAD Analysis

    Directory of Open Access Journals (Sweden)

    Florina Bunghez

    2013-11-01

    Full Text Available Abstract. It is known for a long time that seasoning/condimentary herbs have antioxidant activity and antibacterial properties, being good natural alternatives for disease prevention. The different efficiency of these plants is assigned to their bioactive molecules, stability and bioavailability. In the present study seven aromatic herbs (basil, thyme, oregano, rosemary, clove, cinnamon and sage were investigated individually. A new product was developed using basil, thyme, oregano, rosemary, clove, cinnamon and sage, according to a default recipe. The characterization of each plant aimed to identify the specific “fingerprint” by its main bioactive molecules and the “traceability” of these molecules in the new product, made by mixing the selected plants according to a default recipe. In order to determine the main bioactive compounds of the individual plants composition, in comparison with the new plant-based (EPC formula, high throughput techniques like UV-Vis spectroscopy and LC-QTOF-MS  spectrometry were used. The most important bioactive compounds determined in the studied herbs, which may exert antioxidant activity and antibacterial properties, were phenolic compounds (phenolic acids, flavonoids, quinones, clorophylls as well some polar terpenoids. The fingerprints are providing comprehensive and accurate information about the compounds that may exert antimicrobial properties. In order to assure the biological effects and the bioavailability of the polyphenols and the secondary metabolites we have to consider the antagonistic and synergistic effect that the metabolites can exert on each other.

  1. Stability studies of potent opioid analgesic, morphine-6-O-sulfate in various buffers and biological matrices by HPLC-DAD analysis.

    Science.gov (United States)

    Yadlapalli, Jai Shankar K; Albayati, Zaineb A F; Penthala, Narasimha R; Hendrickson, Howard P; Crooks, Peter A

    2017-09-01

    The 6-O-sulfate ester of morphine (M6S) has previously been shown to be an analgesic with greater potency and fewer side effects than morphine. However, being a sulfate ester derivative of morphine, the question exists as to whether this compound is stable in biological fluids and tissues with regard to pH- and esterase-mediated degradation. To date, no studies have focused on the stability profile of M6S across the physiologically relevant pH range of 1.2-7.4. In addition, the stability of M6S is not known in rat plasma and rat brain homogenate, or in simulated rat gastric and intestinal fluids. This study determines the stability profile of M6S (utilized as the sodium salt) and demonstrates that M6S is highly stable and resilient to either enzymatic- or pH-dependent hydrolysis in vitro. Copyright © 2017 John Wiley & Sons, Ltd.

  2. A HPLC-DAD method for the simultaneous determination of five marker components in the traditional herbal medicine Bangpungtongsung-san

    Directory of Open Access Journals (Sweden)

    Jin Bae Weon

    2011-01-01

    Full Text Available Background: Bangpungtongsung-san, one of the traditional herbal medicines, was known to be a prescription for obesity. Objective: For the simultaneous determination of five components (paeoniflorin, 6-gingerol, decursin, geniposide, and glycyrrhizin in Bangpungtongsung-san, a high-performance liquid chromatography with diode-array detector method was established. Materials and Methods: To develop the method, a reverse phase column, DIONEX C 18 (5 μm, 120 Å, 4.6 mm × 150 mm was used. The mobile phase consisted of methanol and water using a gradient elution. The UV wavelength was set at 230, 240, and 254 nm. Method validation was accomplished by linearity, precision test, and recovery test. Results: All calibration curves of components showed good linearity (R 2 > 0.9959. The limit of detection (LOD and limit of quantification (LOQ ranged from 0.01 to 0.17 μg/ml and 0.04 to 0.53 μg/ml, respectively. The relative standard deviations (RSD value of precision test, intraday and interday tests were less than 0.43% and 1.26%. In the recovery test, results of accuracy ranged from 95.27% to 107.70% with RSD values less than 2.21%. Conclusion: This developed method was applied to the commercial Bangpungtongsung-san sample and the five marker components were separated effectively without interference of any peaks of components.

  3. MULTIRESIDUE DETERMINATION OF ACIDIC PESTICIDES IN WATER BY HPLC/DAD WITH CONFIRMATION BY GC/MS USING CONVERSION TO THE METHYL ESTER WITH TRIMETHYLSILYDIAZOMETHANE

    Science.gov (United States)

    A multiresidue pesticide methodology has been studied and results for acidics are reported here with base/neutral to follow. This work studies a literature procedure as a possible general approach to many pesticides and potentially other analytes that are considered to be liquid...

  4. Analysis of Flavone C-Glycosides in the Leaves of Clinacanthus nutans (Burm. f.) Lindau by HPTLC and HPLC-UV/DAD

    OpenAIRE

    June Lee Chelyn; Maizatul Hasyima Omar; Nor Syaidatul Akmal Mohd Yousof; Ramesh Ranggasamy; Mohd Isa Wasiman; Zakiah Ismail

    2014-01-01

    Clinacanthus nutans (family Acanthaceae) has been used for the treatment of inflammation and herpes viral infection. Currently, there has not been any report on the qualitative and quantitative determination of the chemical markers in the leaves of C. nutans. The C-glycosidic flavones such as shaftoside, isoorientin, orientin, isovitexin, and vitexin have been found to be major flavonoids in the leaves of this plant. Therefore, we had developed a two-step method using thin-layer chromatograph...

  5. Determination of Diphenylamine in Agricultural Products by HPLC-FL.

    Science.gov (United States)

    Aoyagi, Mitsutoshi; Chiba, Masahiro; Kakimoto, Youichiro; Nemoto, Satoru

    2016-01-01

    A method for the determination of diphenylamine in agricultural products was developed. Diphenylamine was extracted with acetonitrile from a sample under an acidic condition, passed through a C18 cartridge column, re-extracted with n-hexane, cleaned up on a PSA cartridge column, determined by HPLC with fluorescence detector and confirmed by liquid chromatography with tandem mass spectrometry. Average recoveries (n=5) from brown rice, corn, soybeans, potato, cabbage, eggplant, spinach, orange, apple and green tea were in the range from 76.7 to 94.9%, and the relative standard deviations were from 0.6 to 5.8% at concentrations equal to the maximum residue limits (MRLs). The quantification limits were 0.01 mg/kg, which is the uniform limit in the positive list system for agricultural chemical residues in food in Japan.

  6. Mean fluorescence lifetime and its error

    Energy Technology Data Exchange (ETDEWEB)

    Fiserova, Eva [Department of Mathematical Analysis and Applications of Mathematics, Faculty of Science, Palacky University in Olomouc, tr. 17. listopadu 12, CZE-77146 Olomouc (Czech Republic); Kubala, Martin, E-mail: mkubala@prfnw.upol.cz [Department of Biophysics, Faculty of Science, Palacky University in Olomouc, tr. 17. listopadu 12, CZE-77146 Olomouc (Czech Republic)

    2012-08-15

    Mean excited-state lifetime is one of the fundamental fluorescence characteristics and enters as an important parameter into numerous calculations characterizing molecular interactions, such as e.g. FRET or fluorescence quenching. Our experiments demonstrated that the intensity-weighted mean fluorescence lifetime is very robust characteristic, in contrast to the amplitude-weighted one, which value is dependent on the data quality and particularly on the used fitting model. For the first time, we also report the procedure for the error estimation for both the intensity- and amplitude-weighted mean fluorescence lifetimes. Furthermore, we present a method for estimation of the mean fluorescence lifetime directly from the fluorescence-decay curve recorded by TCSPC (Time-Correlated Single-Photon Counting) method. For its simplicity and low computational demands, it could be a useful tool in the high-throughput applications, such as FACS, FLIM-FRET or HPLC detectors. - Highlights: Black-Right-Pointing-Pointer Intensity-weighted mean fluorescence lifetime is very robust characteristic. Black-Right-Pointing-Pointer The amplitude-weighted mean lifetime depends on the selection of fitting model. Black-Right-Pointing-Pointer Rigorous procedure for estimation of confidence intervals for mean lifetime. Black-Right-Pointing-Pointer The mean lifetime can be estimated directly from the TCSPC histogram.

  7. Simultaneous quantification of Amaryllidaceae alkaloids from Zephyranthes grandiflora by UPLC-DAD/ESI-MS/MS.

    Science.gov (United States)

    Katoch, Deepali; Kumar, Shiv; Kumar, Neeraj; Singh, Bikram

    2012-12-01

    A rapid, simple and sensitive ultra performance liquid chromatography-diode array detection method (UPLC-DAD) was developed and validated for quantification of four biologically important Amaryllidaceae alkaloids viz. lycoramine, hamayne, haemanthamine and tortuosine in Zephyranthes grandiflora. The method employed BEH C(18) column (2.1mm×100mm, 1.7μm particle size) with linear gradient elution of acetonitrile and water (0.05% formic acid) in a flow rate of 0.3mL/min and at λ(max) 280nm. Standard calibration curve for the analytes were linear (r(2)≥0.9999), precise (intra-day RSDsalkaloids in rainy season. The method was also applied for identification of Amaryllidaceae alkaloids in the plant and overall, seventeen Amaryllidaceae alkaloids of different structural types lycorine, haemanthamine, galanthamine, narciclasine were characterised. This study provides a qualitative and quantitative method for analysis of Amaryllidaceae alkaloids.

  8. Contribution to the characterization of Opuntia spp. juices by LC-DAD-ESI-MS/MS.

    Science.gov (United States)

    Mata, A; Ferreira, J P; Semedo, C; Serra, T; Duarte, C M M; Bronze, M R

    2016-11-01

    Opuntia spp. fruits are considered as health promoting foods due to the diversity of bioactive molecules found in these fruits. The composition in organic acids, flavonols and betalains in the Opuntia ficus-indica juice from a region of Portugal was accomplished for the first time by liquid chromatography and tandem mass spectrometry using an electrospray ionization source operating in negative and positive mode. The methodology used allowed the detection of 44 compounds, from which 32 were identified. Isorhamnetin derivatives were the dominant flavonol glycosides. A total of 9 betalains including 6 betaxanthins and 3 betacyanin were also detected in the fruit juice samples and indicaxanthin, betanin and isobetanin were the major pigments. Phenolic acid and phenylpyruvic acid derivatives were also identified. To our knowledge, it is the first time derivative compounds from piscidic acid, phenolic compounds and betalains are characterized in cactus pear juice using a single LC-DAD-ESI-MS/MS method.

  9. La presencia dadá y surrealista en La tremenda corte

    OpenAIRE

    Ruiz, Blanca E.

    1995-01-01

    Se analizan los rasgos del dadaísmo y del surrealismo en el discurso del programa radiofónico humorístico La tremenda corte, producido en Cuba. En referencia al dadá, el programa hace alusión al absurdo en el sarcasmo en contra de lo convencional y lo solemne, en la construcción anárquica y burlesca de diálogos, así como en el carácter irreverente. El surrealismo se presenta con el uso de sueños como justificantes de los absurdos en el programa, en la presencia en diálogos de un fluir de cons...

  10. LC-DAD-MS-based metabolite profiling of three species of Justicia (Acanthaceae).

    Science.gov (United States)

    Calderón, Angela I; Hodel, Adam; Wolfender, Jean-Luc; Gupta, Mahabir P; Correa, Mireya; Hostettmann, Kurt

    2013-08-01

    Olean-12-en-3β-24 diol (A), auranamide (B), aurantiamide acetate (C), 2α,3β-dihydroxy-olean-12-en-28-oic acid (D) and quindoline (E) were isolated from the dichloromethane (CH2Cl2) extract of the stems of Justicia secunda (Acanthaceae). Liquid chromatography with ultraviolet and mass spectrometric detection was used to acquire more knowledge of the chemical composition of this extract and to monitor variations in profiles of both the isolated and the other non-identified compounds in Justicia refractifolia and Justicia graciliflora. The compound classes, phenolic and olefinic amides, feruloyltyramine amides, 2,5-diaryl-3,4-dimethyltetrahydrofuranoid lignans, peptide alkaloids, phenylalanine derivatives, conjugated ynones, indolquinoline alkaloids, triterpenes and pigments, were tentatively identified based on the LC-DAD-APCI-MS analysis. The most frequently encountered compound among the species was auranamide while the distribution of quindoline was limited to J. secunda. Moreover, the acetylcholinesterase inhibitory activity of the isolated compounds was determined.

  11. Semi-preparative HPLC preparation and HPTLC quantification of tetrahydroamentoflavone as marker in Semecarpus anacardium and its polyherbal formulations.

    Science.gov (United States)

    Aravind, S G; Arimboor, Ranjith; Rangan, Meena; Madhavan, Soumya N; Arumughan, C

    2008-11-04

    Application of modern scientific knowledge coupled with sensitive analytical technique is important for the quality evaluation and standardization of polyherbal formulations. Semecarpus anacardium, an important medicinal plant with wide medicinal properties, is frequently used in a large number of traditional herbal preparations. Tetrahydroamentoflavone (THA), a major bioactive biflavonoid was selected as a chemical marker of S. anacardium and RP-semi-preparative HPLC conditions were optimized for the isolation of tetrahydroamentoflavone. HPTLC analytical method was developed for the fingerprinting of S. anacardium flavonoids and quantification of tetrahydroamentoflavone. The method was validated in terms of their linearity, LOD, LOQ, precision and accuracy and compared with RP-HPLC-DAD method. The methods were demonstrated for the chemical fingerprinting of S. anacardium plant parts and some commercial polyherbal formulations and the amount of tetrahydroamentoflavone was quantified. HPTLC analysis showed that S. anacardium seed contained approximately 10 g kg(-1) of tetrahydroamentoflavone. The methods were able to identify and quantify tetrahydroamentoflavone from complex mixtures of phytochemicals and could be extended to the marker-based standardization of polyherbal formulations, containing S. anacardium.

  12. Stability-Indicating HPLC-UV Method for Vitamin D3 Determination in Solutions, Nutritional Supplements and Pharmaceuticals.

    Science.gov (United States)

    Temova, Žane; Roškar, Robert

    2016-08-01

    A simple and fast high-performance liquid chromatography method with UV detection for determination of vitamin D3 in stability studies as well as in solutions, nutritional supplements and pharmaceuticals was developed. Successful separation of vitamin D3 from its degradation products was achieved on a Gemini C18 100 × 3.0 mm column using a mixture of acetonitrile and water (99:1, v/v) as а mobile phase. The method was successfully validated according to the ICH guidelines. The described reversed-phase HPLC method is favorable compared with other published HPLC-UV methods because of its stability-indicating nature, short run time (3.3 min) and wide analytical range with outstanding linearity, accuracy and precision. The method was further applied for quantification of vitamin D3 in selected liquid and solid nutritional supplements and prescription medicines, confirming its suitability for routine analysis. Degradation products, formed under stress conditions (hydrolysis, oxidation, photolysis and thermal degradation), were additionally elucidated by suitable equipment (LC-DAD-MS) to confirm the stability-indicating nature of the developed method.

  13. HPLC Analysis of Phenolics Compounds and Antioxidant Capacity of Leaves of Vitex megapotamica (Sprengel Moldenke

    Directory of Open Access Journals (Sweden)

    Félix Alexandre Antunes Soares

    2013-07-01

    Full Text Available Vitex megapotamica (Sprengel Moldenke belongs to the Verbenaceae family and is popularly known as “tarumã”. The antioxidant capacity of fractions and crude extract from the leaves of V. megapotamica were determined in this study through the capacity to remove reactive species and phenolic compounds were quantified in the various fractions. The IC50 (DPPH ranged from 14.17 ± 0.76 to 37.63 ± 0.98 µg/mL. The ethyl acetate fraction might contain the strongest lipid peroxidation inhibitory compounds with an IC50 of 16.36 ± 5.09 µg/mL, being also the one with the highest content of polyphenols (522.4 ± 1.12 mg/g, flavonoids (220.48 ± 0.30 mg/g and condensed tannins (3.86 ± 0.53 mg/g. Compounds quantified by HPLC/DAD in the crude extract and fractions were chlorogenic and rosmarinic acids. Higher dosages of the extracts were more effective in reducing levels of plasma protein carbonyls and were also shown to be able to remove reactive species by a 2',7'-dichlorofluorescein diacetate assay, reducing oxidative stress in all tested fractions. Results obtained indicated that V. megapotamica exhibits good potential to prevent diseases caused by the overproduction of free radicals and it might also be used as a potential source of natural antioxidant agents.

  14. HPLC analysis of phenolics compounds and antioxidant capacity of leaves of Vitex megapotamica (Sprengel) Moldenke.

    Science.gov (United States)

    de Brum, Thiele Faccim; Zadra, Marina; Piana, Mariana; Boligon, Aline Augusti; Fröhlich, Janaina Kieling; de Freitas, Robson Borba; Stefanello, Sílvio Terra; Froeder, Amanda Luana Forbrig; Belke, Bianca Vargas; Nunes, Letícia Teixeira; da Silva Jesus, Roberta; Machado, Michel Mansur; da Rocha, João Batista Teixeira; Soares, Félix Alexandre Antunes; Athayde, Margareth Linde

    2013-07-16

    Vitex megapotamica (Sprengel) Moldenke belongs to the Verbenaceae family and is popularly known as "tarumã". The antioxidant capacity of fractions and crude extract from the leaves of V. megapotamica were determined in this study through the capacity to remove reactive species and phenolic compounds were quantified in the various fractions. The IC50 (DPPH) ranged from 14.17 ± 0.76 to 37.63 ± 0.98 µg/mL. The ethyl acetate fraction might contain the strongest lipid peroxidation inhibitory compounds with an IC50 of 16.36 ± 5.09 µg/mL, being also the one with the highest content of polyphenols (522.4 ± 1.12 mg/g), flavonoids (220.48 ± 0.30 mg/g) and condensed tannins (3.86 ± 0.53 mg/g). Compounds quantified by HPLC/DAD in the crude extract and fractions were chlorogenic and rosmarinic acids. Higher dosages of the extracts were more effective in reducing levels of plasma protein carbonyls and were also shown to be able to remove reactive species by a 2',7'-dichlorofluorescein diacetate assay, reducing oxidative stress in all tested fractions. Results obtained indicated that V. megapotamica exhibits good potential to prevent diseases caused by the overproduction of free radicals and it might also be used as a potential source of natural antioxidant agents.

  15. Stability indicating RP-HPLC method for simultaneous determination of piroxicam and ofloxacin in binary combination.

    Science.gov (United States)

    John, Peter; Azeem, Waqar; Ashfaq, Muhammad; Khan, Islam Ullah; Razzaq, Syed Naeem

    2015-09-01

    A simple and precise RP-HPLC method was developed for simultaneous determination of piroxicam and ofloxacin in pharmaceutical formulations and human serum. Optimum separations of piroxicam, ofloxacin and stress-induced degradation products were achieved by use of Hypersil BDS C8 column (250 x 4.6mm, 5 μm). The mobile phase was a mixture of acetonitrile: 0.012M K2HPO4: 0.008M sodium citrate (both buffers mixed and pH adjusted to 2.8) (50:25:25 v/v/v) delivered at flow rate of 1.5 mL min⁻¹ using DAD at 254 nm. Response was linear function of concentration over the ranges of 70-130 mg mL⁻¹ for piroxicam and ofloxacin (r² ≥ 0.999). The method efficiently separated the analytical peaks from degradation products with acceptable tailing and resolution. The developed method was successfully used for concurrent analysis of piroxicam and ofloxacin in pharmaceutical formulations, human serum and in vitro drug interaction studies.

  16. Correlation of Monoclonal Fluorescence Polarization Immunoassay and HPLC for Determination of Whole-Blood Cyciosporin A Concentration%单克隆荧光偏振免疫法和高效液相色谱法测定环孢素A全血浓度的相关性

    Institute of Scientific and Technical Information of China (English)

    戴志凌; 帅莉; 苏琴

    2008-01-01

    目的 比较单克隆荧光偏振免疫法(FPIA-m法)和高效液相色谱(HPLC)法测定环孢素A(CsA)全血浓度的相关性.方法 分别采用HPLC法和FPIA-m法测定42份肾移植患者的CsA血样,测定值用t检验和线性回归方法进行分析.结果 FPIA-m法与HPLC法测定值之间相关性好;回归方程为CF=78.12+1.21 CH,r=0.989 3(P<0.001).结论 FPIA-m法和HPLC法同属于特异性分析法,FPIA-m法测定值均高于HPLC法.

  17. Pronase E Digestion of N-Glycans in Glycoprotein and Its Fluorescent Derivatives Analysis by HPLC-ESI/MS%Pronase E酶解释放糖蛋白N-糖链的方法及荧光标记衍生物的LC-MS分析

    Institute of Scientific and Technical Information of China (English)

    徐莎; 张萍; 黄琳娟; 王仲孚

    2010-01-01

    建立了一种用非特异性酶链酶蛋白酶 E(Pronase E)从糖蛋白上释放N-糖链的方法. 以牛胰核糖核酸酶 B(Ribo B)和鸡白蛋白(Chicken Albumin)为材料,用Pronase E代替N-糖苷酶 F(PNGase F)释放N-糖链. 当蛋白酶质量与糖蛋白质量比为1∶1时,得到只带一个天冬氨酸(Asn)的闭环N-糖链,称其为糖氨酸(glycan-Asn),这样既为糖链引入了天然的-NH2活性基团,同时还保持了糖链原有的还原端闭环结构. 以9-氯甲酸芴甲酯(Fmoc-Cl)为衍生试剂对解离后的糖氨酸进行衍生,采用高效液相色谱-电喷雾质谱联用技术(HPLC-ESI/MS)对Fmoc-Cl糖氨酸衍生物进行分析,建立了糖蛋白的Pronase E酶解、微量糖氨酸的Fmoc-Cl衍生以及糖氨酸衍生物的HPLC-ESI/MS分析方法,该方法保持了N-糖链的天然结构,便于以-NH2为功能基团进一步进行荧光标记、分离制备以及糖链与蛋白质的相互作用研究.

  18. Determination of melamine in milk products by HPLC%高效液相色谱法测定乳制品中的三聚氰胺

    Institute of Scientific and Technical Information of China (English)

    白静

    2011-01-01

    A high performance liquid chromatography method for the determination of meiamine in milk products has been developed. Melamine was determined by using C18 and strong cation mixture filled chromatographic column and DAD detector. The pretreatment of the method was simple, and no ion-pair reagents were needed in mobile phase. The method is proved to be simple, rapid, and accurate for the detection of melamine in milk products.%采用C18和强阳离子交换剂混合填料色谱柱、二极管阵列检测器(DAD),建立一种高效液相色谱法( HPLC)检测乳制品中三聚氰胺的方法.该法前处理简单,且无需在流动枢中添加离子对试剂,具有操作简单、快速、准确性好的特点.

  19. Determination of Anthracycline Drug Residual in Cleaning Validation Swabs of Stainless-Steel Equipment after Production of Cytostatic Injections Using HPLC Analytical Method

    Directory of Open Access Journals (Sweden)

    Zuzana Slivová

    2015-01-01

    Full Text Available Standard cleaning procedures of production line equipment were verified after manufacture of cytostatic injections containing Anthracycline derivate substance. Residual content of Anthracycline drug substance on stainless-steel equipment surface was determined using swab sampling with a specific HPLC-DAD analysis. The acceptance limit was decided as 200.0 μg/100 cm2. Recovery from the stainless-steel surface was 90.1%. Linearity of the method was observed in the concentration range of 0.155–194 μg/mL when estimated using Zorbax TMS (5 μm, 0.25 m × 4.6 mm ID column at 1.3 mL/min flow rate and 254 nm (DAD 190–600 nm. The mobile phase consisted of lauryl hydrogen sulphate solution (3.7 g/L : methanol : acetonitrile (54 : 16 : 30, v/v/v with pH adjusted to 2.5 using phosphoric acid (85%. The LOD and LOQ for Anthracycline derivate were found to be 0.047 and 0.155 μg/mL, respectively. The method validation confirmed the method provides acceptable degree of selectivity, linearity, accuracy, and precision for the intended purposes.

  20. Photoreaction of indole-containing mycotoxins to fluorescent products.

    Science.gov (United States)

    Maragos, C M

    2009-06-01

    Photochemical reaction of the non-fluorescent mycotoxin cyclopiazonic acid (CPA) to fluorescent products was recently reported. Because CPA contains an indole moiety, believed to contribute to the fluorescence, it was of interest to determine whether the effect might be more generally applicable to indole-containing mycotoxins. Three indole-containing tremorgens (penitrem A, paxilline, verruculogen) that have not previously been reported to be fluorescent were rendered fluorescent by exposure to ultraviolet light in a photoreactor. Naturally fluorescent ergot alkaloids, which also contain an indole-moiety, exhibited a diminished response after exposure. This suggests that the phenomenon may be most useful for detection of indole-containing tremorgens that are non-fluorescent, rather than for the enhancement of materials that are already fluorescent, such as the ergot alkaloids. The extent to which fluorescence enhancement was seen was strongly influenced by the reaction environment, in particular the solvent used and whether cyclodextrins were present. In an HPLC format, placement of the photoreactor post-column allowed for the fluorescence detection of penitrem A, paxilline, and verruculogen. The ability to photoreact indole-containing tremorgens and detect them by fluorescence may open up new avenues for detection of these mycotoxins alone or in combination.