Pub Date : 2021-09-03DOI: 10.26420/austinjanalpharmchem.2021.1134
Asghar Mn, Bajwa Aa, A. A., M. A, S. A., S. Sm
Polyphenol Oxidase (PPO) catalyses the undesirable browning of wheat products which is of significant concern in consumer acceptance perspectives. Another important yield-limiting cause for wheat crops is wheat rust (e.g., yellow rust), a source of great economic loss worldwide. The purpose of the current research was to screen conventional and synthetic bread wheat genotypes for their PPO activity and yellow rust resistance. Different genotypes differed significantly in total PPO activity and in their activities against different substrates (L-DOPA and Catechol). The synthetically derived bread wheat genotypes 1-279, showed the lowest (39.2 units/min/g) cumulative PPO activity. Ten genotypes each with the highest and lowest PPO activities were selected for testing their association with seven reported molecular markers. Intriguingly the PPO markers reported in literature could not clearly differentiate between contrasting cultivars. Association with yellow rust resistance was also investigated. Interestingly, the rust-resistant genotypes, including 1-263, Ch-43, 1-57 and Emat (all synthetic-derived), exhibited low PPO activity. The current study underpins that there is a need to search for more reliable PPO markers and to further validate association of low PPO activity with yellow rust.
{"title":"Association of Polyphenol Oxidase Activities with Molecular Markers and Yellow Rust Resistance in Diverse Bread Wheat Genotypes","authors":"Asghar Mn, Bajwa Aa, A. A., M. A, S. A., S. Sm","doi":"10.26420/austinjanalpharmchem.2021.1134","DOIUrl":"https://doi.org/10.26420/austinjanalpharmchem.2021.1134","url":null,"abstract":"Polyphenol Oxidase (PPO) catalyses the undesirable browning of wheat products which is of significant concern in consumer acceptance perspectives. Another important yield-limiting cause for wheat crops is wheat rust (e.g., yellow rust), a source of great economic loss worldwide. The purpose of the current research was to screen conventional and synthetic bread wheat genotypes for their PPO activity and yellow rust resistance. Different genotypes differed significantly in total PPO activity and in their activities against different substrates (L-DOPA and Catechol). The synthetically derived bread wheat genotypes 1-279, showed the lowest (39.2 units/min/g) cumulative PPO activity. Ten genotypes each with the highest and lowest PPO activities were selected for testing their association with seven reported molecular markers. Intriguingly the PPO markers reported in literature could not clearly differentiate between contrasting cultivars. Association with yellow rust resistance was also investigated. Interestingly, the rust-resistant genotypes, including 1-263, Ch-43, 1-57 and Emat (all synthetic-derived), exhibited low PPO activity. The current study underpins that there is a need to search for more reliable PPO markers and to further validate association of low PPO activity with yellow rust.","PeriodicalId":91055,"journal":{"name":"Austin journal of analytical and pharmaceutical chemistry","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2021-09-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42776708","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2021-08-07DOI: 10.26420/austinjanalpharmchem.2021.1133
Yu J, M. B, L. J., Chen Y, W. Z, C. J, D. Y
Chicken spleen Transfer Factor (TF) is a low-molecular-weight lymphocyte extract composed of polypeptide and nucleotide. However, its role in regulating intestinal structure and function in laying hens has remained largely unknown. 100 one-day-old laying hens were randomly divided into five groups and administered with different doses of TF (0.00 [control], 0.05mL, 0.10mL, 0.25mL and 1.00mL). The results showed that the high dose of TF (1.00mL) improved the intestinal mucosa morphology and strengthened the digestive and absorption function. Furthermore, the histology analysis revealed an increase in the number of intraepithelial lymphocytes and goblet cells. Similarly, the results from ELISA demonstrated an increase in the content of IL-10 in the intestinal tract, while the content of TNF-a showed a decrease in this regard. The RT-PCR assay also demonstrated the upregulation of the relative mRNA expressions of Muc2, TLR-2, and TLR-4 genes. The intestinal antioxidant function was significantly enhanced. In conclusion, high-dose of TF can improve the intestinal mucosa morphology and structure, enhance digestion and absorption functions, enhance the intestinal mucosal barrier immune function and antioxidant function, and up-regulate Muc2, TLR-2 and TLR-4 gene relative expression.
{"title":"The Effect of Chicken Spleen Transfer Factor on Intestinal Mucosa Immunity Barrier of Laying Hens","authors":"Yu J, M. B, L. J., Chen Y, W. Z, C. J, D. Y","doi":"10.26420/austinjanalpharmchem.2021.1133","DOIUrl":"https://doi.org/10.26420/austinjanalpharmchem.2021.1133","url":null,"abstract":"Chicken spleen Transfer Factor (TF) is a low-molecular-weight lymphocyte extract composed of polypeptide and nucleotide. However, its role in regulating intestinal structure and function in laying hens has remained largely unknown. 100 one-day-old laying hens were randomly divided into five groups and administered with different doses of TF (0.00 [control], 0.05mL, 0.10mL, 0.25mL and 1.00mL). The results showed that the high dose of TF (1.00mL) improved the intestinal mucosa morphology and strengthened the digestive and absorption function. Furthermore, the histology analysis revealed an increase in the number of intraepithelial lymphocytes and goblet cells. Similarly, the results from ELISA demonstrated an increase in the content of IL-10 in the intestinal tract, while the content of TNF-a showed a decrease in this regard. The RT-PCR assay also demonstrated the upregulation of the relative mRNA expressions of Muc2, TLR-2, and TLR-4 genes. The intestinal antioxidant function was significantly enhanced. In conclusion, high-dose of TF can improve the intestinal mucosa morphology and structure, enhance digestion and absorption functions, enhance the intestinal mucosal barrier immune function and antioxidant function, and up-regulate Muc2, TLR-2 and TLR-4 gene relative expression.","PeriodicalId":91055,"journal":{"name":"Austin journal of analytical and pharmaceutical chemistry","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2021-08-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43641845","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2021-05-05DOI: 10.26420/AUSTINJANALPHARMCHEM.2021.1132
Elizariev An, Zaki Eg, el-Kousy Sm
Recently a new virus strain designated as SARS coronavirus result in a fatal pandemic known as COVID-19. Bioinformatics and drug screening are directed for the assessment of potential inhibitors before their clinical implementation for the treatment of this fatal pneumonia. One of the expected natural potent inhibitors is Portulaca oleracea which has been assigned as an effective drug to different human ailments throughout the whole world. P. oleracea is widely spread in most areas of Egypt. In the current study, hydrophilic polysaccharides were purified from Portulaca oleracea extracts. Molecular docking simulation is implemented to investigate the antiviral effect of the purified polysaccharides to inhibit COVID-19. The viral protease was downloaded from a Protein Data Bank (PDB# 6y84) then docked with the potent inhibitors. The docking results indicate that the purified polysaccharides can bind tightly to the SARS-CoV-2 viral protease, which indicates that P. oleracea is a potential inhibitor for COVID-19.
{"title":"Investigating the Inhibition Effect of Portulaca oleracea against SARS-CoV-2 through Molecular Docking Simulation","authors":"Elizariev An, Zaki Eg, el-Kousy Sm","doi":"10.26420/AUSTINJANALPHARMCHEM.2021.1132","DOIUrl":"https://doi.org/10.26420/AUSTINJANALPHARMCHEM.2021.1132","url":null,"abstract":"Recently a new virus strain designated as SARS coronavirus result in a fatal pandemic known as COVID-19. Bioinformatics and drug screening are directed for the assessment of potential inhibitors before their clinical implementation for the treatment of this fatal pneumonia. One of the expected natural potent inhibitors is Portulaca oleracea which has been assigned as an effective drug to different human ailments throughout the whole world. P. oleracea is widely spread in most areas of Egypt. In the current study, hydrophilic polysaccharides were purified from Portulaca oleracea extracts. Molecular docking simulation is implemented to investigate the antiviral effect of the purified polysaccharides to inhibit COVID-19. The viral protease was downloaded from a Protein Data Bank (PDB# 6y84) then docked with the potent inhibitors. The docking results indicate that the purified polysaccharides can bind tightly to the SARS-CoV-2 viral protease, which indicates that P. oleracea is a potential inhibitor for COVID-19.","PeriodicalId":91055,"journal":{"name":"Austin journal of analytical and pharmaceutical chemistry","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2021-05-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42414193","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2021-03-10DOI: 10.26420/AUSTINJANALPHARMCHEM.2021.1131
H. Salem, Omar Ma, Derayea Sm, Khalil Aa
A comparative study for the validation and advancement of two analytical approaches applied for the simultaneous determination of Mometasone Furoate (MF), Miconazole Nitrate (MIC) and Gentamicin (GM) formulated in Momenta® cream. The first approach was TLC-spectrodensitometric method, which was advanced by separating the three components on TLC aluminum plates coated with silica gel 60 F254 using chloroform: methanol: formic acid (4:0.3:0.15, v/v/v) as a mobile phase, then scanned at 254nm using Camage TLC scanner 3 operated in reflectance-absorbance mode. The second approach was the chemometric method using two models: Partial Least Squares (PLS) and Principle Component Regression Model (PCR). The proposed approaches were validated according to ICH guidelines and were applied for the determination of the ternary mixtures in their analytical mixtures and pharmaceutical preparation.
{"title":"Application of TLC-Spectrodensitometric and Chemometric Methods for Determination of Momenta® Cream: A Comparative Study Applied on Ternary Mixture","authors":"H. Salem, Omar Ma, Derayea Sm, Khalil Aa","doi":"10.26420/AUSTINJANALPHARMCHEM.2021.1131","DOIUrl":"https://doi.org/10.26420/AUSTINJANALPHARMCHEM.2021.1131","url":null,"abstract":"A comparative study for the validation and advancement of two analytical approaches applied for the simultaneous determination of Mometasone Furoate (MF), Miconazole Nitrate (MIC) and Gentamicin (GM) formulated in Momenta® cream. The first approach was TLC-spectrodensitometric method, which was advanced by separating the three components on TLC aluminum plates coated with silica gel 60 F254 using chloroform: methanol: formic acid (4:0.3:0.15, v/v/v) as a mobile phase, then scanned at 254nm using Camage TLC scanner 3 operated in reflectance-absorbance mode. The second approach was the chemometric method using two models: Partial Least Squares (PLS) and Principle Component Regression Model (PCR). The proposed approaches were validated according to ICH guidelines and were applied for the determination of the ternary mixtures in their analytical mixtures and pharmaceutical preparation.","PeriodicalId":91055,"journal":{"name":"Austin journal of analytical and pharmaceutical chemistry","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2021-03-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45702170","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2021-03-03DOI: 10.26420/AUSTININTERNMED.2021.1129
N. Singh, Akhtar Mj, A. Anchliya
The objective of this study was the development, optimization, and validation of a RP-HPLC method for the quantification of reduced glutathione (GSH) and oxidized glutathione (GSSG) in pharmaceutical formulations The separation utilized a C18 column at room temperature with absorption wavelength 210nm. The mobile phase was an isocratic flow of a 95:5 (v/v) mixture of 25mM phosphate buffer (pH 2.7) and methanol with flow rate at 1.0 mL/min. Validation of the method assessed with the methods ability in seven categories: linearity, range, limit of detection, limit of quantification, accuracy, precision, and selectivity. The method show an acceptable degree of linearity with r²=0.9994 and 0.999 over a concentration range of 10-200 μg/mL for GSH and GSSG respectively. The detection limit and quantification limit for GSH 20.7μg/mL and 69.24μg/mL and for GSSG 17.22μg/mL and 57.42μg/mL respectively. The percent recovery of the method was 99.98-100.93 %. Following validation, the method was employed in the determination of glutathione in pharmaceutical formulations in the form of a liposome. The proposed method offers a simple, accurate, and inexpensive way to quantify reduced glutathione.
{"title":"Development and Validation of HPLC Method for Simultaneous Estimation of Reduced and Oxidized Glutathione in Bulk Pharmaceutical Formulation","authors":"N. Singh, Akhtar Mj, A. Anchliya","doi":"10.26420/AUSTININTERNMED.2021.1129","DOIUrl":"https://doi.org/10.26420/AUSTININTERNMED.2021.1129","url":null,"abstract":"The objective of this study was the development, optimization, and validation of a RP-HPLC method for the quantification of reduced glutathione (GSH) and oxidized glutathione (GSSG) in pharmaceutical formulations The separation utilized a C18 column at room temperature with absorption wavelength 210nm. The mobile phase was an isocratic flow of a 95:5 (v/v) mixture of 25mM phosphate buffer (pH 2.7) and methanol with flow rate at 1.0 mL/min. Validation of the method assessed with the methods ability in seven categories: linearity, range, limit of detection, limit of quantification, accuracy, precision, and selectivity. The method show an acceptable degree of linearity with r²=0.9994 and 0.999 over a concentration range of 10-200 μg/mL for GSH and GSSG respectively. The detection limit and quantification limit for GSH 20.7μg/mL and 69.24μg/mL and for GSSG 17.22μg/mL and 57.42μg/mL respectively. The percent recovery of the method was 99.98-100.93 %. Following validation, the method was employed in the determination of glutathione in pharmaceutical formulations in the form of a liposome. The proposed method offers a simple, accurate, and inexpensive way to quantify reduced glutathione.","PeriodicalId":91055,"journal":{"name":"Austin journal of analytical and pharmaceutical chemistry","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2021-03-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47797398","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2021-03-03DOI: 10.26420/AUSTININTERNMED.2021.1128
K. Verma, Akhtar Mj, A. Anchliya
Fourier transform infrared spectroscopy is an effectual and noncritical approach for quantitative study of different types analytes present in pharmaceutical and foodstuffs. The present review describes the basic principles and the instrumentation of FTIR spectroscopy along with its sample preparation techniques, sample handling techniques and advancements. FTIR spectroscopy in combination with chemometrics techniques has been followed over long times. The main objective of this review is to assemble the data linked to application of FTIR spectroscopic and chemometrics techniques for the quantitative study of varieties of analytes like API, adulterants, caffeine, cocaine, lipids, fats & oils, sugar and others. The FTIR spectroscopy with chemometrics techniques proved to be a beneficial methodology for quantitative study to routine analysis of these analytes.
{"title":"Combination of FTIR Spectroscopy and Chemometric Method on Quantitative Approach - A Review","authors":"K. Verma, Akhtar Mj, A. Anchliya","doi":"10.26420/AUSTININTERNMED.2021.1128","DOIUrl":"https://doi.org/10.26420/AUSTININTERNMED.2021.1128","url":null,"abstract":"Fourier transform infrared spectroscopy is an effectual and noncritical approach for quantitative study of different types analytes present in pharmaceutical and foodstuffs. The present review describes the basic principles and the instrumentation of FTIR spectroscopy along with its sample preparation techniques, sample handling techniques and advancements. FTIR spectroscopy in combination with chemometrics techniques has been followed over long times. The main objective of this review is to assemble the data linked to application of FTIR spectroscopic and chemometrics techniques for the quantitative study of varieties of analytes like API, adulterants, caffeine, cocaine, lipids, fats & oils, sugar and others. The FTIR spectroscopy with chemometrics techniques proved to be a beneficial methodology for quantitative study to routine analysis of these analytes.","PeriodicalId":91055,"journal":{"name":"Austin journal of analytical and pharmaceutical chemistry","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2021-03-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49225497","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2021-03-03DOI: 10.26420/AUSTININTERNMED.2021.1130
M. Hashemi, Hadjmohammadi Mr
This paper discusses the recent development of enantioselective extraction methods. There are several methods in this field. The principles of all of enantioselective extraction methods are presented a focus on twophase extractions, in these methods used a chiral compound that named chiral selector, this compound distinguishes between two chiral isomers in complexation process, then one isomer more captured by chiral selector, and separation processes happen. The chiral selector should dissolve in on phase. In these methods, typical performance data are reported major emphasis on distribution ratio and enantioselectivity value or selectivity factor. In this paper, we introduce types of enantioselective extraction method, and discuss about them with several examples.
{"title":"Recent Development: Enantioselective Extraction in Chiral Separation","authors":"M. Hashemi, Hadjmohammadi Mr","doi":"10.26420/AUSTININTERNMED.2021.1130","DOIUrl":"https://doi.org/10.26420/AUSTININTERNMED.2021.1130","url":null,"abstract":"This paper discusses the recent development of enantioselective extraction methods. There are several methods in this field. The principles of all of enantioselective extraction methods are presented a focus on twophase extractions, in these methods used a chiral compound that named chiral selector, this compound distinguishes between two chiral isomers in complexation process, then one isomer more captured by chiral selector, and separation processes happen. The chiral selector should dissolve in on phase. In these methods, typical performance data are reported major emphasis on distribution ratio and enantioselectivity value or selectivity factor. In this paper, we introduce types of enantioselective extraction method, and discuss about them with several examples.","PeriodicalId":91055,"journal":{"name":"Austin journal of analytical and pharmaceutical chemistry","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2021-03-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42862078","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2021-01-25DOI: 10.26420/austinjanalpharmchem.2022.1139
Gaafar A, Ayyad R
This search includes the vitamin B-species which must be taken by diabetic patients. Where these vitamins keep the health of patient not merely but help the metabolism of glucose which is the blood sugar. The vitamin B-complex must be prescribed before the drugs which treat the diabetes (Insulin and oral hypoglycaemics). Because these vitamins keep the integrity of patient health and play the major role of metabolism of glucose.
{"title":"Important of B-Complex Vitamins in Treatment Protocol in Diabetic Patients","authors":"Gaafar A, Ayyad R","doi":"10.26420/austinjanalpharmchem.2022.1139","DOIUrl":"https://doi.org/10.26420/austinjanalpharmchem.2022.1139","url":null,"abstract":"This search includes the vitamin B-species which must be taken by diabetic patients. Where these vitamins keep the health of patient not merely but help the metabolism of glucose which is the blood sugar. The vitamin B-complex must be prescribed before the drugs which treat the diabetes (Insulin and oral hypoglycaemics). Because these vitamins keep the integrity of patient health and play the major role of metabolism of glucose.","PeriodicalId":91055,"journal":{"name":"Austin journal of analytical and pharmaceutical chemistry","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2021-01-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47238118","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Warfarin is an oral anticoagulant that requires frequent therapeutic drug monitoring due to a narrow therapeutic window, considerable interindividual variability in drug response, and susceptibility to drug-drug and drug-diet interactions. Enantiomeric separation and quantification of warfarin enantiomers and clinically important major hydroxylation metabolites are essential for drug interaction studies and phenotypic characterization of CYP2C9 and CYP3A4, the major cytochrome P450 (CYP) enzymes involved in warfarin metabolism. Here, we describe the development and validation of a chiral high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS)-based quantification of R-warfarin, S-warfarin, S-7-hydroxywarfarin (the major CYP2C9 metabolite) and (9R;10S)-10-hydroxywarfarin (the CYP3A4 metabolite) in human plasma. Simple protein precipitation-based extraction showed good recovery of analytes (82.9 - 96.9%). The developed method exhibited satisfactory intra-day and inter-day accuracy and precision. The lower limits of detection were 0.25 nM (or ~0.08 ng/mL) for the warfarin enantiomers and 0.1 nM (or ~0.04 ng/mL) for S-7-hydroxywarfarin and (9R;10S)-10-hydroxywarfarin using only 50 µL plasma during extraction. The validated method was successfully applied to analyze plasma samples obtained from a healthy human subject who enrolled in a clinical drug interaction study involving warfarin.
{"title":"A chiral HPLC-MS/MS method for simultaneous quantification of warfarin enantiomers and its major hydroxylation metabolites of CYP2C9 and CYP3A4 in human plasma.","authors":"W Ju, K Peng, S Yang, H Sun, M Sampson, M Z Wang","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>Warfarin is an oral anticoagulant that requires frequent therapeutic drug monitoring due to a narrow therapeutic window, considerable interindividual variability in drug response, and susceptibility to drug-drug and drug-diet interactions. Enantiomeric separation and quantification of warfarin enantiomers and clinically important major hydroxylation metabolites are essential for drug interaction studies and phenotypic characterization of CYP2C9 and CYP3A4, the major cytochrome P450 (CYP) enzymes involved in warfarin metabolism. Here, we describe the development and validation of a chiral high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS)-based quantification of <i>R</i>-warfarin, <i>S</i>-warfarin, <i>S</i>-7-hydroxywarfarin (the major CYP2C9 metabolite) and (9<i>R</i>;10<i>S</i>)-10-hydroxywarfarin (the CYP3A4 metabolite) in human plasma. Simple protein precipitation-based extraction showed good recovery of analytes (82.9 - 96.9%). The developed method exhibited satisfactory intra-day and inter-day accuracy and precision. The lower limits of detection were 0.25 nM (or ~0.08 ng/mL) for the warfarin enantiomers and 0.1 nM (or ~0.04 ng/mL) for <i>S</i>-7-hydroxywarfarin and (9<i>R</i>;10<i>S</i>)-10-hydroxywarfarin using only 50 µL plasma during extraction. The validated method was successfully applied to analyze plasma samples obtained from a healthy human subject who enrolled in a clinical drug interaction study involving warfarin.</p>","PeriodicalId":91055,"journal":{"name":"Austin journal of analytical and pharmaceutical chemistry","volume":"1 2","pages":""},"PeriodicalIF":0.0,"publicationDate":"2014-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4494745/pdf/nihms693615.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"33893181","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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