Determination of progesterone in rat plasma by LC-MS/MS / 国际药学研究杂志

ABSTRACT

Objective: To develop and validate an LC-MS/MS method for determination of progesterone in rat plasma. Methods: Plasma samples were prepared by liquid-liquid extraction with a mixture of n-hexane/dichloromethane/isopropanol (100505, V/V/V). Isocratic chromatographic separation was performed on a Hypersil ODS column (50 mm × 2.1 mm, 5 μm, Thermo). The mobile phase was methanol-water (7525, V/V). Detection of progesterone and the internal standard (IS) megestrol acetate was achieved by ESI MS/MS in the positive ion mode using m/z 315.4→ m/z(97.0 + 109.2) and m/z 385.4→ m/z 267.4 transitions, respectively. Results: The method was linear from 0.5 to 200 ng/ml when 100 μl plasma was analyzed. The lower limit of quantification was 0.5 ng/ml. Both the inter- and intra-day precision values were below 6.7%, and the accuracy (relative error) was within ±6.2% in all quality control samples. Conclusion: The method provides a specific, sensitive and rapid means for the determination of progesterone in rat plasma, and completely meets the requirements for pharmacokinetic study.