Development of a sensitive method for simultaneous determination of fluticasone propionate and salmeterol in plasma samples by liquid chromatography‐tandem mass spectrometry

Abstract

A new method for the fast simultaneous quantification of fluticasone propionate and salmeterol from plasma samples by liquid chromatography-tandem mass spectrometry, with adequate sensitivity for pharmacokinetic applications, was developed and validated. The chromatographic separation and mass-spectrometric parameters were optimized for the retention and detection of the two compounds, despite quite different structures and properties. Two columns connected in series were used, cation-exchange (Zorbax 300-SCX, 5 cm x 2.1 mm, 5 μm) and octadecyl (Discovery HSC₁₈, 10 cm x 2.1 mm, 5 μm). The mass-spectrometric interface was operated in negative electrospray ionization mode; high sensitivity and lesser matrix effects were obtained, permitting smaller consumption of plasma. The sample preparation was based on supported liquid-liquid extraction in 96-well format plates that provided clean samples with a simplified procedure that was suitable for automation. The method was validated according to regulatory guidelines, by assessing lower limits of quantification, selectivity, linearity, accuracy, precision, extraction recoveries and matrix effects. A comparison with two other methods for the separate determination of fluticasone propionate and salmeterol in plasma samples, previously developed by our group, is presented. The statistical evaluation of the results obtained with the three methods on a set of unknown samples from treated patients demonstrated good correlation (R² 0.987 for fluticasone propionate and 0.967 for salmeterol).

Population pharmacokinetics of fluticasone propionate/salmeterol using two different dry powder inhalers

“…The lower limits of quantification were 1.500 pg/mL and 2.500 pg/mL for SAL and FLP, respectively (Silvestro et al, 2012).…”

Pharmacokinetic analysis of inhaled salmeterol in asthma patients: Evidence from two dry powder inhalers

“…Separations were performed on a reversed phase column Ascentis Phenyl, 10 cm × 2.1 mm, 5 μm (Merck), in isocratic conditions using a mobile phase composition of 90% acetonitrile and 10% ammonium acetate 15 m m in water, with a flow rate of 1 ml/min. The utilized technique was a re‐validated version of the previously published method (Silvestro, Savu, Savu, Tudoroniu, & Tarcomnicu, ). The lower limit of quantification for salmeterol was 1.00 pg/ml.…”

Determination of salmeterol, α‐hydroxysalmeterol and fluticasone propionate in human urine and plasma for doping control using UHPLC–QTOF–MS

“…The stability of salmeterol and fluticasone propionate in plasma and urine has been evaluated in previous papers and shown to be relatively stable. Both compounds are stable in plasma for 4 months at −20 ο C (Silvestro et al, 2012). β-2-Agonists are stable in urine for 30 days at −20 ο C (Wang et al, 2015).…”