中国临床药学杂志

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(75 mm×2.0 mm,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(R_s,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。

AIM To develop a sensitive and accurate method for the simultaneous determination 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 cleaned by Waters Oasis^? HLB solid-phase extraction cartridge.The separation was performed on a Shim-pack XR-ODSⅡcolumn(75 mm×2.0 mm,2.2μm) with a linear gradient elution program using acetonitrile (containing 0.1%formic acid) and 0.1%formic acid aqueous solution as mobile phase.Electrospray ionization was applied and operated in the positive ion multiple reaction monitoring(MRM) mode.The quantitation was carried on by methyltestosterone-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,R_S,N≥10) were in the range of 0.5 -1.0μg·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 were all lower than 10.0%.CONCLUSION The developed method is specific,sensitive and simple,and is conformed to the requirements of biological specimen analyses.