Determination of cortisol production rates with contemporary liquid chromatography-mass spectrometry to measure cortisol-d3 dilution after infusion of deuterated tracer
Determination of cortisol production rates with contemporary liquid chromatography-mass spectrometry to measure cortisol-d3 dilution after infusion of deuterated tracer
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DOI：
10.1016/j.clinbiochem.2010.12.008
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发表时间：
2011-04-01
影响因子：
2.8
通讯作者：
DeBarber, Andrea E.
中科院分区：
文献类型：
--
作者：
Klopfenstein, Bethany J.;Purnell, Jonathan Q.;DeBarber, Andrea E.
目的：通过稳态输注氘化皮质醇和质谱分析稳定同位素稀释来测量24小时皮质醇生成率（CPR）是检测人类下丘脑-垂体-肾上腺轴活性的一种有价值的工具。我们已经开发并验证了一种改进的方法，用现代LC-MS技术测量皮质醇稀释度。设计与方法：血浆样品和校准器用乙酸乙酯提取。LC-MS采用配备常压化学电离（APCI）源的Surveyor HPLC和TSQ Quantum三重四极杆质谱仪。结果：通过消除被鉴定为20 β -二氢皮质醇的干扰，选择性比以前的方法有所提高。血浆校准器在1.5 ~ 10%皮质醇-d(3)浓度范围内线性关系良好，相关系数> 0.995。结论：该APCI LC-MS方法简化了样品的检查和分析，并可选择性检测低浓度的皮质醇-d(3)输注，用于测定24 h CPR。(C) 2011加拿大临床化学学会。Elsevier Inc.出版。版权所有。
Objectives: Measurement of 24-h cortisol production rate (CPR) using steady-state infusion of deuterated cortisol and analysis of stable-isotope dilution by MS is a valuable tool to examine hypothalamic-pituitary-adrenal axis activity in humans. We have developed and validated an improved method for measuring cortisol dilution with contemporary LC-MS technology.Design and methods: Plasma samples and calibrators were extracted with ethylacetate. LC-MS was performed with a Surveyor HPLC and TSQ Quantum triple-quadrupole mass spectrometer equipped with an atmospheric pressure chemical ionization (APCI) source.Results: Selectivity was improved over previous methods via elimination of an interferent identified as 20 beta-dihydrocortisol. The LLOQ for cortisol-d(3) was 2.73 nmol/L and LOD 1.37 nmol/L Plasma calibrators were linear over the concentration range 1.5-10% cortisol-d(3), with correlation coefficients > 0.995.Conclusions: This APCI LC-MS method offers simplified sample work-up and analysis and enables selective detection of the low concentration of cortisol-d(3) infused for determination of 24-h CPR. (C) 2011 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.