title: Development and validation of a highly sensitive LC-MS/MS-ESI method for quantification of IIIM-019-A novel nitroimidazole derivative with promising action against Tuberculosis: Application to drug development.
abstract: The study aims to illustrate an analytical validation of a rapid and sensitive liquid chromatography (LC) coupled to tandem mass spectrometry (MS-MS) and electrospray ionization (ESI) method for quantification of IIIM-019 (a novel nitroimidazole derivative with potential activity against Tuberculosis) in mice plasma. The extraction of the analyte and the internal standard (Tolbutamide) from the plasma samples involves protein precipitation using acetonitrile. The chromatographic separation was accomplished using a gradient mode and the mobile phase comprised of acetonitrile and 0.1% formic acid in water. The flow rate used was 0.7 ml/min on a C18e high performance Chromolith column. IIIM-019 and Tolbutamide (IS) were analyzed by combined reversed-phase LC/MS-MS with positive ion electrospray ionization. The MS-MS ion transitions used were 533>170.1, 533>198 for IIIM-019 and 271>74, 271>155 for internal standard (IS) respectively. The method was linear over a concentration range of 0.5-1000 ng/ml and the lower limit of quantification was 0.50 ng/ml. The entire study was validated for accuracy, precision, linearity, range, selectivity, lower limit of quantification (LLOQ), recovery, and matrix effect in accordance with the FDA guidelines of method validation. Acceptable precision and accuracy were obtained for concentrations over the standard curve range. The intra and inter-day precisions were in the range of 0.51-11.18% and 0.51-7.55%. The pharmacokinetics was performed on male Balb/c mice by oral (2.5mg/kg), intraperitoneal (2.5mg/kg) and intravenous (1mg/kg) routes. The oral bioavailability of IIIM-019 was 51.6%. The method was also applied successfully in determining microsomal stability wherein the compound was found to be very slightly metabolized by rat liver microsomes.
authors.affiliation: Inflammation Pharmacology Division, CSIR-Indian Institute of Integrative Medicine, Canal Road, Jammu 180001, J&K, India; Academy of Scientific and Innovative Research, CSIR-Indian Institute of Integrative Medicine, Canal Road, Jammu 180001, J&K, India.
authors.forename: Gurleen
authors.initials: G
authors.lastname: Kour
authors.affiliation: Inflammation Pharmacology Division, CSIR-Indian Institute of Integrative Medicine, Canal Road, Jammu 180001, J&K, India.
authors.forename: Bal Krishan
authors.initials: BK
authors.lastname: Chandan
authors.affiliation: Inflammation Pharmacology Division, CSIR-Indian Institute of Integrative Medicine, Canal Road, Jammu 180001, J&K, India; Academy of Scientific and Innovative Research, CSIR-Indian Institute of Integrative Medicine, Canal Road, Jammu 180001, J&K, India.
authors.forename: Mowkshi
authors.initials: M
authors.lastname: Khullar
authors.affiliation: Academy of Scientific and Innovative Research, CSIR-Indian Institute of Integrative Medicine, Canal Road, Jammu 180001, J&K, India; Medicinal Chemistry Division, CSIR-Indian Institute of Integrative Medicine, Canal Road, Jammu 180001, J&K, India.
authors.forename: Gurunadham
authors.initials: G
authors.lastname: Munagala
authors.affiliation: Medicinal Chemistry Division, CSIR-Indian Institute of Integrative Medicine, Canal Road, Jammu 180001, J&K, India.
authors.forename: Parvinder Pal
authors.initials: PP
authors.lastname: Singh
authors.affiliation: Inflammation Pharmacology Division, CSIR-Indian Institute of Integrative Medicine, Canal Road, Jammu 180001, J&K, India.
authors.forename: Asha
authors.initials: A
authors.lastname: Bhagat
authors.affiliation: Quality Control and Quality Analysis Division, CSIR-Indian Institute of Integrative Medicine, Canal Road, Jammu 180001, J&K, India.
authors.forename: Ajai Prakash
authors.initials: AP
authors.lastname: Gupta
authors.affiliation: Director- Indian Institute of Integrative Medicine, anal Road, Jammu 180001, J&K, India.
authors.forename: Ram A
authors.initials: RA
authors.lastname: Vishwakarma
authors.affiliation: Inflammation Pharmacology Division, CSIR-Indian Institute of Integrative Medicine, Canal Road, Jammu 180001, J&K, India. Electronic address: zahmed@iiim.ac.in.
authors.forename: Zabeer
authors.initials: Z
authors.lastname: Ahmed
mesh_terms: D000818:Animals; D000995:Antitubercular Agents / Q000032:analysis* / Q000493:pharmacokinetics / Q000494:pharmacology; D002853:Chromatography, Liquid / Q000379:methods*; D015195:Drug Design; D057230:Limit of Detection; D008862:Microsomes, Liver / Q000378:metabolism; D009593:Nitroimidazoles / Q000032:analysis* / Q000493:pharmacokinetics / Q000494:pharmacology; D051381:Rats; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D014376:Tuberculosis / Q000517:prevention & control*
chemical_list: D000995:Antitubercular Agents; D009593:Nitroimidazoles
keywords: IIIM-019; LC–MS/MS; Microsomal stability; Nitroimidazole; Pharmacokinetic study