Parallel diurnal fluctuation of testosterone, androstenedione, dehydroepiandrosterone and 17OHprogesterone as assessed in serum and saliva: validation of a novel liquid chromatography-tandem mass spectrometry method for salivary steroid profiling
Parallel diurnal fluctuation of testosterone, androstenedione, dehydroepiandrosterone and 17OHprogesterone as assessed in serum and saliva: validation of a novel liquid chromatography-tandem mass spectrometry method for salivary steroid profiling
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DOI：
10.1515/cclm-2016-0805
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发表时间：
2017-09-01
影响因子：
6.8
通讯作者：
Fanelli, Flaminia
中科院分区：
文献类型：
--
作者：
Mezzullo, Marco;Fazzini, Alessia;Fanelli, Flaminia
背景：唾液雄激素检测是一种有价值的生物学信息来源。然而，这种低水平的适当测量对于直接免疫测定是具有挑战性的，缺乏足够的准确性。在过去的几年里，许多相互矛盾的结果报告低相关性与血清同行阻碍了唾液雄激素检测的临床应用。液相色谱-串联质谱方法：从500 μ L唾液中提取睾酮（T）、雄烯二酮（A）、脱氢表雄酮（DHEA）和17羟孕酮（17 OHP），用9.5 min液相色谱梯度分离，用正离子电喷雾电离-多反应监测法检测。唾液和血清雄激素的日变化由四对收集协议描述19名健康受试者分别于上午8时、中午12时、下午4时和下午8时进行了上述4个时间点的动态观察。该测定允许T、A、DHEA和17 OHP的定量分别降至3.40、6.81、271.0和23.7 pmol/L，所有分析物的准确度在83.0和106.1%之间。所有雄激素在唾液和血清中均观察到平行的昼夜节律，从早晨到晚上时间点，数值逐渐降低。唾液雄激素水平与血清雄激素水平呈高度线性相关（T：R > 0.85; A：R > 0.90; DHEA：R > 0.73和17 OHP：R > 0.89;所有p < 0.0001）。我们的LC-MS/MS方法允许对雄激素唾液水平进行敏感的评价，并且代表了探索综合性研究的相关性的最佳技术。在唾液中测量雄激素谱，用于研究生理和病理状态下的雄激素分泌调节和活性。
Background: Salivary androgen testing represents a valuable source of biological information. However, the proper measurement of such low levels is challenging for direct immunoassays, lacking adequate accuracy. In the last few years, many conflicting findings reporting low correlation with the serum counterparts have hampered the clinical application of salivary androgen testing. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) makes it possible to overcome previous analytical limits, providing new insights in endocrinology practice.Methods: Salivary testosterone (T), androstenedione (A), dehydroepiandrosterone (DHEA) and 17OHprogesterone (17OHP) were extracted from 500 mu L of saliva, separated in 9.5 min LC-gradient and detected by positive electrospray ionization-multiple reaction monitoring. The diurnal variation of salivary and serum androgens was described by a four paired collection protocol (8 am, 12 am, 4 pm and 8 pm) in 19 healthy subjects.Results: The assay allowed the quantitation of T, A, DHEA and 17OHP down to 3.40, 6.81, 271.0 and 23.7 pmol/L, respectively, with accuracy between 83.0 and 106.1% for all analytes. A parallel diurnal rhythm in saliva and serum was observed for all androgens, with values decreasing from the morning to the evening time points. Salivary androgen levels revealed a high linear correlation with serum counterparts in both sexes (T: R > 0.85; A: R > 0.90; DHEA: R > 0.73 and 17OHP: R > 0.89; p < 0.0001 for all).Conclusions: Our LC-MS/MS method allowed a sensitive evaluation of androgen salivary levels and represents an optimal technique to explore the relevance of a comprehensive androgen profile as measured in saliva for the study of androgen secretion modulation and activity in physiologic and pathologic states.