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%.

Key words

antidiabetic - LC-MS/MS - S007-1261 - pharmacokinetics - rat plasma

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