Salivary testosterone measurement by liquid chromatography tandem mass spectrometry in adult males and females.
Salivary testosterone measurement by liquid chromatography tandem mass spectrometry in adult males and females.
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DOI：
10.1177/0004563213506412
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发表时间：
2014-05
影响因子：
2.2
通讯作者：
NATSAL Team
中科院分区：
文献类型：
--
作者：
Keevil BG;MacDonald P;Macdowall W;Lee DM;Wu FC;NATSAL Team
唾液睾酮(SAL-T)可能是血清游离睾酮的有效替代物。本研究的目的是使用一种新的LC-MS/MS分析方法来确定SAL-T浓度是否能准确地反映性别间的SAL-T浓度，并探讨样本采集的实用方面。采集了104名男性和91名女性的唾液和血清样本。建立了一种更灵敏的LC-MS/MS分析方法，使在女性体内发现的低浓度SAL-T得以定量。唾液(200μL)加入D5-睾酮后，用1mL甲基叔丁基醚提取。使用Waters TQ-S质谱仪进行定量。该方法的定量下限为5pmol/L，足够灵敏地测定女性唾液中的睾酮。SAL-T表现出日变化，但每周和每月的采样没有显着差异。经冻融保存3年后，SAL-T在常温下最长可保存5天。SAL-T的参考区间男性为93~378pmol/L，女性为5~46pmoL/L。男性SAL-T与计算的游离T显著相关(P<0.001)，女性与SA-T显著相关(r=0.39，P<；0.001)。这些结果证实了LC-MS/MS法可以可靠、准确地测定成年男性和女性唾液中的睾酮。这些结果为进一步探索SAL-T作为血清睾酮的可靠替代品在雄激素紊乱的诊断和治疗中的临床应用以及在临床研究中评估雄激素状态奠定了基础。
Salivary testosterone (Sal-T) may be a useful surrogate of serum free testosterone. The study aims were to use a novel liquid chromatography tandem mass spectrometry (LC-MS/MS) assay to determine whether Sal-T concentrations accurately reflect Sal-T concentrations in both sexes and to investigate practical aspects of sample collection. Saliva and serum samples were collected in 104 male and 91 female subjects. A more sensitive LC-MS/MS assay was developed to enable Sal-T quantitation in the low concentrations found in females. Saliva (200 μL) was extracted with 1 mL of methyl-tert-butyl ether following the addition of D5-testosterone. Quantitation was performed using a Waters TQ-S mass spectrometer. The assay achieved a lower limit of quantification of 5pmol/L, sufficiently sensitive to measure testosterone in female saliva. Sal-T showed a diurnal variation but samples taken at weekly and monthly intervals showed no significant differences. Sal-T was stable at ambient temperature for up to 5 days, after freeze-thawing and 3 years frozen storage. Reference intervals for Sal-T were 93-378 pmol/L in males and 5-46 pmol/L in females. Sal-T correlated significantly with serum calculated free-T in males (r=0.71, P<0.001) and in females (r=0.39, P<0.001). These results confirm that testosterone can be reliably and accurately measured by LC-MS/MS in both adult male and female saliva samples. These results lay the foundation for further exploration of the clinical application of Sal- T as a reliable alternative to serum testosterone in the diagnosis and management of androgen disorders and assessment of androgen status in clinical research.