Development and validation of liquid chromatography-tandem mass spectrometric method for the quantification of ciprofibrate from human plasma.

Abstract

A new rapid, selective and sensitive liquid chromatography-tandem mass spectrometric method was developed and validated for the determination of ciprofibrate, an antihyperlipidemic agent, in K2EDTA human plasma. Furosemide was used as internal standard (IS). The ciprofibrate and IS were extracted using Oasis HLB 1 cc 30 mg solid-phase extraction cartridge. The chromatographic separation was performed on ACE C18, 50 × 4.6 mm, 5 µm column. The mobile phase consisted of 0.001% ammonia in methanol-acetonitrile-water (70:20:10, v/v/v). Detection and quantitation were performed by a triple quadrupole equipped with electrospray ionization and multiple reaction monitoring in negative ionization mode. The most intense [M-H](-) transition for ciprofibrate at m/z 287.0 → 85.0 and for IS at m/z 328.9.0 → 204.9 were used for quantification. The method was found to linear over the range of 25-30,000 ng/mL (r > 0.998). The lower limit of quantitation (LLOQ) was 25 ng/mL. The extraction recovery was above 90%. The accuracy was found to be 101.26-106.44%. The stability testing was also investigated and it was found that both drug and IS were quite stable. The developed method was successfully applied to the bioequivalence study of ciprofibrate 100 mg tablet after oral administration to healthy human volunteers.