Development and validation of a sensitive LC-MS/MS method for simultaneous quantitation of fluticasone and salmeterol in human plasma

Abstract

A simple, robust bioanalytical method (BM) was developed and validated for simultaneous quantification of fluticasone (FCS) and salmeterol (SMR) in biological matrix (BM). The solid phase extraction (SPE) procedure was use to extract FCS and SMR from BM. The developed method was validated as per regulatory guidelines with calibration range of FCS and SMR with were 0.500-150.200pg/mL and 1.00-250.218pg/mL. The mass transaction ions for FCS and SMR were 501.300→293.000, 415.900→232.200 with column Discovery C18, gradient mobile phase 0.1mM ammonium trifluoro acetate, acetonitrile with injection volume 15µL. The linear regression 1/X² was confirmed with three precision and accuracy (P&A) batches and the best r value among three P&A was 0.9998. The developed BM was showed no carry over, free of inter conversion between FCS & SMR and no matrix effect. The extraction procedure showed more than 80% recovery from spiked samples. The stabilities of FCS and SMR in aqueous and matrix were met the acceptance criteria as per guidelines. The stability in aqueous samples was 36hrs at 2-8℃ and stability in matrix was up to 36 days at -70℃ and -20℃.In conclusion, developed BM will useful in pharmacokinetic studies of FCS and SMR either individual and combined pharmaceutical products.