Simultaneous measurement of cortisol and cortisone in human saliva using liquid chromatography-tandem mass spectrometry: Application in basal and stimulated conditions
Simultaneous measurement of cortisol and cortisone in human saliva using liquid chromatography-tandem mass spectrometry: Application in basal and stimulated conditions
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DOI：
10.1016/j.jchromb.2009.09.014
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发表时间：
2009-11-01
影响因子：
3
通讯作者：
Keevil, Brian G.
中科院分区：
文献类型：
--
作者：
Perogamvros, Ilias;Owen, Laura J.;Keevil, Brian G.
用于测量唾液皮质醇的免疫测定受到可的松可变干扰的限制。唾液可的松是唾液腺表达11 β -羟基类固醇脱氢酶2型（11 β - hsd2）的结果，该酶能将皮质醇转化为可的松。我们报道了唾液皮质醇和可的松（分别为SalF和SalE）液相色谱-串联质谱（LC-MS/MS）联合检测，以解决可的松在皮质醇免疫分析中的交叉反应性，并作为研究11 β - hsd2活性的工具。SalF和SalE在400 nmol/L和200 nmol/L的浓度范围内呈线性关系，定量下限分别为0.39 nmol/L和0.78 nmol/L。没有发现离子抑制的证据，精密度、准确度和回收率在国际公认的限度内。13种结构相关类固醇未发现干扰。在刺激肾上腺后，早晨的SalF、SalE和SalF/SalE显著高于睡前。随着血清皮质醇的升高，SalF呈指数上升，SalE呈线性上升，并在较高的SalF浓度下达到平台。我们开发了一种新的，强大的LC-MS/MS分析方法，用于SalF和SalE的联合测量。我们的研究结果证实了唾液腺的11 β - hsd2活性导致高SalE浓度和高底物浓度下的酶饱和。该方法可作为一种简单、无创和高度特异性的工具来评估唾液皮质醇作为游离血清皮质醇替代品的价值，并作为评估11 β - hsd2活性的潜在新方法。(C) 2009 Elsevier B.V.版权所有
Immunoassays used for the measurement of salivary cortisol are limited by variable interference from cortisone. Salivary cortisone is a consequence of the salivary glands expressing 11 beta-hydroxysteroid dehydrogenase type 2 (11 beta-HSD2) which converts cortisol to cortisone. We report a combined salivary cortisol and cortisone (SalF and SalE respectively) liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay to address the Cortisone cross-reactivity in cortisol immunoassays and as a tool to study 11 beta-HSD2 activity. The method was linear up to 400 nmol/L for SalF and 200 nmol/L for SalE and the lower limits of quantitation were 0.39 nmol/L (SalF) and 0.78 nmol/L (SalE). No evidence of ion suppression Was found and precision, accuracy and recovery were within internationally accepted limits. No interference was identified from 13 structurally related steroids. SalF, SalE and SalF/SalE were significantly greater in the morning than at bed-time and following stimulation of the adrenal glands. As serum cortisol increased, an exponential rise was observed in SalF and a linear increase in SalE which reached a plateau at higher SalF concentrations. We have developed a novel, robust LC-MS/MS assay for the combined measurement of SalF and SalE. Our results confirm the 11 beta-HSD2 activity of the salivary glands resulting in high SalE concentrations and the enzyme saturation at high Substrate concentrations. This method can be used as a simple, non-invasive and highly specific tool to assess the Value of salivary cortisol as a Surrogate for free serum cortisol and as a potential novel way to assess 11 beta-HSD2 activity. (C) 2009 Elsevier B.V. All rights reserved.