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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-D_3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。
Objective To establish a sensitive and reliable isotope dilution-solid phase extraction coupled with ultra-fast liquid chromatography-tandem mass spectrometry (UFLC-MS / MS) for the simultaneous determination of fluoxymesterone, , The United States androsterone, testosterone, methyltestosterone and androstenedione two kinds of protein assimilation hormones. Methods The plasma samples were precipitated with methanol and purified on a Waters Oasis HLB cartridge using a Shim-pack XR-ODS II column (75 mm × 2.0 mm, 2.2 μm) with acetonitrile (0.1% formic acid) and Water (containing 0.1% formic acid) as the mobile phase gradient elution, electrospray positive ion multiple reaction monitoring (MRM) mode detection, methyltestosterone-D_3 as internal standard, using isotope dilution method. Results The target compounds of the 8 tested compounds had good linearity (r> 0.999) in each concentration range, and the limit of quantification (R_ (s, N)> 10) was 0.5-1.0 μg · L -1 ). The relative recoveries of the methods were 95.0% -105.0%. The intra-day RSDs of three spiked samples with low, medium and high spiked concentrations were all lower than 6.0% and the intraday RSDs were less than 10.0%. Conclusion This method is highly specific, sensitive, easy to operate and meets the requirements of biological sample analysis.