STUDY OF THE IN-VITRO METABOLIC PROFILE OF AMLODIPINE IN HUMAN HEPATIC CELL LINE AND CHICKEN LIVER TISSUE USING LIQUID CHROMATOGRAPHY-MASS SPECTROMETRY/MASS SPECTROMETRY

ABSTRACTObjective: The main objective of this study was to investigate the in-vitro metabolic profile of Amlodipine (AMD) using normal human hepatic celllines and chicken liver tissue and to characterize the metabolites obtained using liquid chromatography-tandem mass spectrometry (LC-MS/MS).Methods: In the present study, the metabolic profile of AMD, a well-known calcium channel blocker, was investigated in normal human hepatic celllines and chicken liver tissue employing LC-MS/MS technique. The structural details on AMD metabolites were acquired using triple quadrupole massspectrometer (LCMS-8040, Shimadzu). The metabolites were produced by incubation of AMD with the human hepatic cell lines and chicken livertissue at 37 °C for 24 h. The incubated extracts were analyzed on LC-MS/MS and their product ion spectra were acquired, interpreted and tentativestructures were proposed.Results: Twelve Phase I and Phase II metabolites were successfully detected in the proposed study. The main metabolic changes observed were oxidativedeamination, N-acetylation, de-esterification, hydrogenation, de-methylation, aliphatic hydroxylation, and glucuronidation of dehydrogenated AMD.Based on this information, the tentative structures of the metabolites were postulated.Conclusion: The in-vitro metabolites of AMD were successfully investigated and characterized in human hepatic cell lines and chicken liver tissue.Furthermore, both models were found to be equally effective for carrying out the in-vitro metabolic study of AMD.Keywords: Amlodipine, Metabolites, Hepatic cell lines, Chicken liver tissue, Liquid chromatography, Liquid chromatography-mass spectrometry/mass spectrometry

INVESTIGATION OF IN VITRO METABOLITES OF ETODOLAC IN HUMAN HEPATIC CELL LINE AND CHICKEN LIVER TISSUE USING LC-MS/MS

Objective: The main objective of this study was to investigate the in vitro metabolic profile of etodolac (ETD) using normal human hepatic cell lines and chicken liver tissue, and to characterize the metabolites obtained using Liquid Chromatography-tandem Mass Spectrometry (LC-MS/MS).Methods: In the present study, the metabolic profile of ETD, a well-known non-steroidal anti-inflammatory drug (NSAID), was investigated in normal human hepatic cell lines and chicken liver tissue employing LC-MS/MS technique. The structural details on ETD metabolites were acquired using triple quadrupole mass spectrometer (LCMS-8040, Shimadzu). The metabolites were produced by incubation of ETD with the human hepatic cell lines and chicken liver tissue, at 37 °C for 24 h. The incubated extracts were analyzed with LC-MS/MS and their production spectra were acquired, interpreted and tentative structures were proposed.Results: Six phase I and phase II metabolites were successfully detected in the proposed study. The metabolic changes observed included-oxidation, N-acetylation, hydrogenation, decarboxylation, methylation and glucuronidation of dehydrogenated ETD. The tentative structures of the metabolites were postulated based on the chemical reactions predicted and the LC-MS/MS data obtained.Conclusion: The in vitro metabolites of ETD were successfully investigated and characterized in human hepatic cell lines and chicken liver tissue. Also, both the models were found to be equally effective for carrying out the in vitro metabolic study of ETD.Keywords: Etodolac, Metabolites, Hepatic cell lines, Chicken liver tissue, LC, LC-MS/M

INVESTIGATION OF FORCED DEGRADATION PRODUCTS OF ETODOLAC BY LC AND LC-MS/MS

Objective: The main objective of this study was to investigate the degradation products (DPs) of Etodolac (ETD) API under different stress conditions (acid hydrolysis, base hydrolysis, oxidation, thermal and photolysis) and to characterize the major DPs using Liquid Chromatography-tandem Mass Spectrometry (LC-MS/MS) and Infrared (IR) spectroscopy.Methods: The proposed study describes a reversed-phase LC and LC-MS/MS method. Separation of ETD and its DPs was achieved on a Phenomenex C18 column. Structures of the major DPs were studied using a Triple Quadrupole Mass Spectrometer. A separate gradient LC-MS/MS method was developed for this purpose. The analysis was done using Shim-pack XR ODS column. Possible chemical reactions were predicted depending on the degradation type and the fragmentation data obtained from LC-MS/MS. Major DPs were isolated using preparative LC technique. These DPs were obtained in solid form using rotavap and lyophilizer and were then analyzed by IR spectroscopy to confirm their structural details.Results: ETD was found to degrade completely under acid hydrolysis; it degraded 68 % under oxidation, 25 % after photolysis, 6 % in basic conditions and less than 1 % in thermal degradation. Four major DPs were characterized using LC-MS/MS and IR spectroscopy.Conclusion: The DPs of ETD were investigated and characterized. Also, the tentative degradation pathways of ETD under different stress conditions were postulated.Keywords: Etodolac, Forced degradation studies, Degradation products, LC, LC-MS/MSÂ