超快速液相色谱-串联质谱法测定血浆中8种蛋白同化激素含量

目的建立一种灵敏、可靠的同位素稀释-固相萃取-超快速液相色谱-串联质谱法（UFLC-MS／MS）同时测定人血浆中氟甲睾酮、群勃龙、勃地酮、诺龙、美雄酮、睾酮、甲睾酮和雄烯二酮等8种蛋白同化激素的方法。方法血浆样品经甲醇沉淀蛋白后，用Waters Oasis HLB固相萃取小柱进行净化，采用Shim-pack XR-ODSⅡ色谱柱（75min×2．0min，2．2μm），以乙腈（含0．1％甲酸）和水（含0．1％甲酸）为流动相进行梯度洗脱，电喷雾正离子多反应监测（MRM）模式下检测，以甲睾酮-D3为内标，采用同位素稀释法定量。结果8种待测目标化合物在各自的浓度范围内有均具有良好的线性（r〉0．999），定量检出限（RS．N〉10）为0．5～1．0μg·L^-1，方法的相对回收率为95．0％-105．0％，低、中、高3个加标浓度样品的日内RSD均〈6．0％，日间RSD均〈10．0％。结论本方法专属性强、灵敏度高，操作简便，符合生物样品分析要求。

Simultaneous determination of eight protein anabolic hormones in human plasma by ultrafast liquid chromatography-tandem mass spectrometry

MM To develop a sensitive and accurate method for the simultaneous detemaination of 8 protein anabolic hormones in human plasma by ultrafast liquid chromatography- tandem mass spectrometry （UFLC-MS/MS）with isotope internal standard dilution technique and solid-phase extraction. METHODS Proteins in plasma sample were precipitated by methanol and eleaned by Waters Oasis HLB solid-phase extraction cartridge. The separation was performed on a Shim-pack XR-ODSⅡ column （75 mm × 2.0 mm, 2.2 μm） with a linear gradient elution program using acetonitrile （containing 0.1% formic acid） and 0.1% formic acid aqueous solution as mobile phase. Electmspray ionization was applied and operated in the positive ion multiple reaction monitoring （MRM） mode. The quantitation was carried on by methyltestostemne-D3 isotope internal standard dilution technique. RESULTS The calibration curves were in good linearity for the 8 analytes in the each linearity range with the correlative coefficients more than 0.999. Limits of quantitation （LOQs, RS.N≥ 10） were in the range of 0.5 - 1.0 μg L^-1, and the relative recoveries were between 95.0% and 105.0%. The intra-day relative standard deviations （RSDs）for assaying the plasma sample containing three concentrations of the 8 analytes were all lower than 6.0%, and inter-day RSDs -sere all lower than 10.0%. CONCLUSION The developed method is specific, sensitive and simple, and is conformed to the requirements of biological specimen analyses.