全自动在线涡流固相萃取-液相色谱-串联质谱法直接分析血清中4种雄激素

摘要

A novel method was developed for the direct analysis of testosterone, androstenedione, methyltestosterone and methenolone in serum samples by fully automated online turbulent flow solid phase extraction coupled with high performance liquid chromatography-tandem mass spectrometry. An aliquot of 50 μL serum sample was preconcentrated directly on a Turboflow SPE column after centrifugation. Turboflow SPE C18-P could be used to remove serum matrix effectively. The optimum loading flow rate and elution time were 4. 0 mL/min and 1. 0 min, respectively. The linearity ranges were from 1. 0 μg/L to 100. 0 μg/L for four target compounds. The method limits of detection ( LODs) were in the range of 0. 2-0. 3 μg/L. The relative standard deviations (RSDs) ranged from 2. 9% to 14. 1% (n=5). The time for one sample analysis including extraction, separation and determination was 32 min. The proposed method has been successfully applied for the analysis of serum samples.%建立了一种全自动在线涡流固相萃取-液相色谱-质谱联用直接分析血清中4种雄激素的分析方法。50μL血清样品经高速离心后,可直接用涡流固相萃取柱在线富集和分析。 Turboflow C18-P在线固相萃取柱可有效去除血清基质。最佳上样流速和洗脱时间分别为4.0 mL/min和1.0 min。方法的线性范围在1.0~100.0μg/L之间,4种雄激素的检出限在0.2~0.3μg/L之间,相对标准偏差为2.9％~14.1％(n=5)。应用本方法对实际血清样品进行了分析,整个分析过程包括提取、分离、检测以及柱子再生在32 min内即可完成。