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近几十年来,丙酸氟替卡松与昔萘酸沙美特罗联合给药广泛应用于哮喘的治疗.本研究的目地是建立一个新颖、灵敏的超高效液相色谱-串联质谱法同时测定人血浆中的丙酸氟替卡松与沙美特罗浓度.样品前处理采用简单的96孔板固相萃取,色谱分离在ACQUITY UPLC BEH C18色谱柱(1.7 μm,50 mm×2.1 mm)上进行,以0.1％的氨水溶液为流动相A,甲醇为流动相B,在0.5 mL/min流速下梯度洗脱.分析物和内标的检测采用正离子电喷雾多反应监测模式,监测离子对为m/z 501.4＞313.2(丙酸氟替卡松),506.4＞293.3(丙酸氟替卡松-d5),416.4＞232.1(沙美特罗)和419.3＞235.2(沙美特罗-d3).两个分析物的线性范围为2.50-500 pg/mL,日内和日间精密度准确度均在8.6％以内.丙酸氟替卡松与沙美特罗提取回收率分别高于51.0％和54.6％,回收率不受浓度影响.该方法的验证遵循FDA、EMA和CFDA(中国食品药品监督管理局)的指导原则,且己成功地应用于中国健康受试者吸入单剂量丙酸氟替卡松与沙美特罗(250μg/50 μg)的临床研究.“,”Combined administration of fluticasone propionate and salmeterol xinofoate has 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).