Abstract:

Combined administration of fluticasone propionate and salmeterol xinofoatehas been widely used for the treatment of asthma in recent decades. In this investigation, we developed and validated a novel and sensitive ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for simultaneous determination of fluticasone propionate and salmeterol xinofoate in human plasma. Following a simple SPE sample extraction in 96-well plate format, chromatography was performed on a Waters ACQUITY UPLC BEH C₁₈ column (1.7 μm, 50 mm×2.1 mm) with mobile phase consisting of 100% MeOH and 0.1% NH₄OH in water on a gradient program at flow rate of 0.5 mL/min. Detection of analytes and internal standards was accomplished using multiple reaction monitoring (MRM) of precursor>product ion pairs of m/z 501.4>313.2(fluticasone propionate), 506.4>293.3 (fluticasone propionate-d5), 416.4>232.1 (salmeterol xinofoate) and 419.3>235.2 (salmeterol-d3). The assay range was 2.50–500 pg/mL for both analytes, and a 1/x² weighted linear regression model was used. The inter-assay accuracy and precision of the method were within ±8.6%. The recoveries from 0.30 mL of plasma were greater than 51.0% and 54.6% for fluticasone propionate and salmeterol, respectively, and the results were consistent across low, middle and high concentration levels. The method was validated following FDA, EMA and CFDA (China Food and Drug Administration)’s guidance on bioanalysis and then successfully applied to support a clinical study in healthy Chinese subjects following inhaled administration of a single combination of fluticasone propionate/salmeterol (250 μg/50 μg).

Key words: Fluticasone propionate, Salmeterol xinofoate, UPLC-MS/MS, Human plasma, Chinese subjects