Quantification of cortisol in human eccrine sweat by liquid chromatography - tandem mass spectrometry.
Quantification of cortisol in human eccrine sweat by liquid chromatography - tandem mass spectrometry.
复制标题
DOI：
10.1039/c5an02387d
复制
发表时间：
2016-03-21
期刊：
影响因子：
--
通讯作者：
Sternberg EM
中科院分区：
文献类型：
--
作者：
Jia M;Chew WM;Feinstein Y;Skeath P;Sternberg EM
长期以来，皮质醇一直被认为是评估压力相关疾病的“压力生物标志物”。血浆、尿液或唾液是目前皮质醇分析的来源。这些生物流体的采样要么是侵入性的，要么存在可靠性问题，可能导致不准确的结果。汗液作为一种有前途的非侵入性应力分析来源，已引起越来越多的关注。建立了测定人汗液中皮质醇（(11β)-11,17,21-三羟基孕酮-4-烯-3,20-二酮）含量的高效液相色谱-质谱联用方法。用混合模式RP - HPLC从皮质醇中分离出至少一种未知的异构体，这种异构体以前没有报道过，可能会干扰定量。以皮质醇-9,11,12,12- d4为内标，采用常压化学电离法（APCI）和正离子模式选择反应监测法（SRM）检测皮质醇。定量限和定量限分别为0.04 ng/ml和0.1 ng/ml。在0.10 ~ 25.00 ng/ml的线性范围内。日内精密度（2.5% ~ 9.7%）、准确度（0.5% ~ 2.1%）、日间精密度（12.3% ~ 18.7%）、准确度（7.1% ~ 15.1%）均可实现。该方法已成功应用于人体汗液样本的皮质醇分析。这是首次证明HPLC-MS/MS可以在至少一种与皮质醇具有相似疏水性的异构体存在的情况下，用于灵敏和高度特异性地测定人体分泌汗液中的皮质醇。这项研究表明，人类汗液可以作为无创评估应激生物标志物（如皮质醇和其他类固醇激素）的有希望的来源。人汗液中皮质醇与其异构体的分离
Cortisol has long been recognized as the “stress biomarker” in evaluating stress related disorders. Plasma, urine or saliva are the current source for cortisol analysis. The sampling of these biofluids is either invasive or has reliability problems that could lead to inaccurate results. Sweat has drawn increasing attention as a promising source for non-invasive stress analysis. A sensitive HPLC-MS/MS method was developed for the quantitation of cortisol ((11β)-11,17,21-Trihydroxypregn-4-ene-3,20-dione) in human eccrine sweat. At least one unknown isomer that has previously not been reported and could potentially interfere with quantification was separated from cortisol with mixed mode RP HPLC. Detection of cortisol was carried out using atmospheric pressure chemical ionization (APCI) and selected reaction monitoring (SRM) in positive ion mode, using cortisol-9,11,12,12-D4 as internal standard. LOD and LOQ were estimated to be 0.04 ng/ml and 0.1 ng/ml, respectively. Linear range of 0.10 – 25.00 ng/ml was obtained. Intraday precision (2.5% – 9.7%) and accuracy (0.5% – 2.1%), interday precision (12.3% – 18.7%) and accuracy (7.1% – 15.1%) were achieved. This method has been successfully applied to the cortisol analysis of human eccrine sweat samples. This is the first demonstration that HPLC-MS/MS can be used for the sensitive and highly specific determination of cortisol in human eccrine sweat in the presence of at least one isomer that has similar hydrophobicity as cortisol. This study demonstrated that human eccrine sweat could be used as a promising source for non-invasive assessment of stress biomarkers such as cortisol and other steroid hormones. Separation of cortisol from its isomers in human eccrine sweat