title: Bioanalytical LC-MS/MS method development and validation of novel antidiabetic candidate S007-1261 in rat plasma and its application to pharmacokinetic and oral bioavailability studies.
abstract: A sensitive and selective LC-MS/MS method has been developed and validated for CDRI antidiabetic candidate S007-1261 in rat plasma using 16-dehydropregnenolone as an internal standard. The API 4000 triple quadrupole LC-MS/MS system was operated under multiple reaction monitoring mode using electrospray ionization technique in positive mode. The sample processing method involves 2-step liquid-liquid extraction using n-hexane as an extracting solvent. The analyte was chromatographed on RP 18, waters column (3.5 µm, 2.1 mm i.d. × 30 mm) with guard using acetonitrile and ammonium acetate buffer (pH 5.0, 10 mM) in 90:10 (v/v) composition at a flow rate of 0.40 mL min(-1). The chromatographic run time was 5.30 min. Calibration curve shows linearity over concentration range 1.56-200 ng mL(-1). The lower limit of detection was 0.39 ng mL(-1) and lower limit of quantitation was 1.56 ng mL(-1). The inter- and intra-day accuracy and precision were found to be within the assay variability limits as per US FDA guidelines. The absolute recovery of S007-1261 was found to be >90%. S007-1261 does not show any stability problems as it was stable at room temperature for 8 h. S007-1261 was also stable up to 3 freeze-thaw cycles and can be stored up to 30 days at -60 °C. The assay was successfully applied to both oral (40 mg kg(-1)) and intravenous (10 mg kg(-1)) pharmacokinetic studies in male Sprague-Dawley rats. The oral bioavailability of S007-1261 was found to be 33.61%.
authors.affiliation: Pharmacokinetics and Metabolism Division, CSIR-Central Drug Research Institute, Lucknow, India.
authors.forename: A
authors.initials: A
authors.lastname: Misra
authors.affiliation: Pharmacokinetics and Metabolism Division, CSIR-Central Drug Research Institute, Lucknow, India.
authors.forename: H N
authors.initials: HN
authors.lastname: Kushwaha
authors.affiliation: Pharmacokinetics and Metabolism Division, CSIR-Central Drug Research Institute, Lucknow, India.
authors.forename: N
authors.initials: N
authors.lastname: Gautam
authors.affiliation: Pharmacokinetics and Metabolism Division, CSIR-Central Drug Research Institute, Lucknow, India.
authors.forename: B
authors.initials: B
authors.lastname: Singh
authors.affiliation: Medicinal and Process Chemistry Division, CSIR-Central Drug Research Institute, Lucknow, India.
authors.forename: P C
authors.initials: PC
authors.lastname: Verma
authors.affiliation: Medicinal and Process Chemistry Division, CSIR-Central Drug Research Institute, Lucknow, India.
authors.forename: R
authors.initials: R
authors.lastname: Pratap
authors.affiliation: Pharmacokinetics and Metabolism Division, CSIR-Central Drug Research Institute, Lucknow, India.
authors.forename: S K
authors.initials: SK
authors.lastname: Singh
mesh_terms: D000284:Administration, Oral; D000818:Animals; D001682:Biological Availability; D002138:Calibration; D006207:Half-Life; D007004:Hypoglycemic Agents / Q000008:administration & dosage / Q000097:blood* / Q000493:pharmacokinetics*; D007275:Injections, Intravenous; D008297:Male; D010091:Oximes / Q000008:administration & dosage / Q000097:blood* / Q000493:pharmacokinetics*; D011283:Pregnenes / Q000008:administration & dosage / Q000097:blood* / Q000493:pharmacokinetics*; D011284:Pregnenolone / Q000031:analogs & derivatives; D011786:Quality Control; D051381:Rats; D017207:Rats, Sprague-Dawley; D012015:Reference Standards; D015203:Reproducibility of Results
chemical_list: D007004:Hypoglycemic Agents; D010091:Oximes; D011283:Pregnenes; C000596971:S007-1261; C003592:16-dehydropregnenolone; D011284:Pregnenolone