A liquid chromatography-tandem mass spectrometry assay for the profiling of classical and 11-oxygenated androgens in saliva
A liquid chromatography-tandem mass spectrometry assay for the profiling of classical and 11-oxygenated androgens in saliva
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DOI：
10.1177/0004563219847498
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发表时间：
2019-09-01
影响因子：
2.2
通讯作者：
Keevil, Brian G.
中科院分区：
文献类型：
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作者：
Schiffer, Lina;Adaway, Joanne E.;Keevil, Brian G.
背景经典雄激素和11-氧合雄激素都对雄激素库有贡献。在类固醇生成障碍中需要定期监测雄激素状态，并且雄激素的多重检测提高了测定的诊断能力。由于廉价的非侵入性收集，当需要多个样本时，唾液是有利的。液相色谱-串联质谱法（LC-MS/MS）可在短运行时间内对类固醇进行灵敏的同时定量。在这里，我们已经开发了一种LC-MS/MS测定同时测量唾液中的17-羟孕酮，雄烯二酮，睾酮，11 β-羟雄烯二酮和11-酮睾酮。方法用二氯甲烷支持液萃取法制备300 μ L非刺激性全唾液，用40%甲醇复溶。用C18柱在线固相萃取后，在C8柱上使用含有0.1%甲酸和2 mmol/L乙酸铵的水/甲醇梯度进行液相色谱。使用沃茨TQ-S质谱仪进行定量。结果总运行时间为6.4 min，回收率在89%~ 109%之间，离子抑制率在86%~ 105%之间。批内和批间比较显示变异系数
Background Classical and 11-oxygenated androgens both contribute to the androgen pool. Regular monitoring of the androgen status is required in disorders of steroidogenesis, and multiplexing of androgens improves the diagnostic ability of an assay. Due to the cheap non-invasive collection, saliva is advantageous when multiple samples are required. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) offers sensitive, simultaneous quantification of steroids with short run times. Here, we have developed an LC-MS/MS assay for the simultaneous measurement of 17-hydroxyprogesterone, androstenedione, testosterone, 11 beta-hydroxyandrostenedione and 11-ketotestosterone in saliva. Methods Samples (300 mu L unstimulated whole saliva) were prepared by supported liquid extraction with dichloromethane and were reconstituted in 40% methanol. After online solid phase extraction with C18 cartridges, liquid chromatography was performed on a C8 column using a water/methanol gradient containing 0.1% formic acid and 2 mmol/L ammonium acetate. A Waters TQ-S mass spectrometer was used for quantification. Results Total run time was 6.4 min. For all analytes, recovery was between 89% and 109%, ion suppression between 86% and 105%. Intra- and inter-assay comparisons showed a coefficient of variation