Bioanalytical Method Development and Validation for a Novel Secretory Phospholipase A₂ Inhibitor by LC-MS/MS and its Application to a Pharmacokinetic Study in Mice

Abstract

Objective: This study aimed to develop and validate a sensitive LC–MS/MS method for quantifying a novel Secretory phospholipase A₂ (sPLA₂) inhibitor, compound-7(L), in mouse plasma and to evaluate its application in pharmacokinetic studies. Methods: A bioanalytical LC–MS/MS method was developed and validated according to ICH M10 guidelines. Compound-7(L) was extracted from mouse plasma using protein precipitation. Pharmacokinetic studies were conducted in male C57BL/6 mice following intravenous administration at a dose of 1 mg/kg. Results: The method was linear over the range of 0.100–100 ng/ml with regression coefficient>0.99. Intra-and Inter-run accuracy ranged from 89.67% to 105.00% and 90.00 % to100%, respectively. The intra-and inter-day precision values were1.88% to 9.52% and 0.30 % to 4.00%, respectively. All stability assessments were within acceptance criteria of ±15%. Conclusion: The method exhibits excellent specificity and selectivity, with well-fitted linearity over the validated concentration range. The established method, validated in line with ICH M10 guidelines, was shown to be accurate, precise, rugged, and stable in mouse plasma, meeting all specified acceptance criteria. Overall, the validated method, owing to its simplicity, sensitivity, and cost‑effectiveness, is suitable for mouse plasma analysis and supports its application in pharmacokinetic study.