UHPLC-MS/MS同时测定人血浆中21-去氧皮质醇、17-羟孕酮、皮质醇和可的松水平的方法建立及临床应用

IF 2.1 Q3 PHARMACOLOGY & PHARMACY
Advances in Pharmacological and Pharmaceutical Sciences Pub Date : 2025-01-17 eCollection Date: 2025-01-01 DOI:10.1155/adpp/3859670
Syed N Alvi, Saleh Al Dgither, Ali Al-Odaib
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引用次数: 0

摘要

采用超高效液相色谱-串联质谱联用技术(UHPLC-MS/MS)建立了一种简便、高效的定量人血浆中21-去氧皮质醇(21-DOC)、17-羟孕酮(17-OHP)、皮质醇和可的松的方法。血浆样品在Atlantis dC18 (3 μm)色谱柱上进行分析,流动相为20.0 mM醋酸铵和乙腈(50:50,v: v),等压流速为0.3 mL/min。加入d4-皮质醇作为内标(IS)后,用二氯甲烷和叔丁基甲基醚1:2 (v/v)的混合物提取含有21-DOC、17-OHP、皮质醇和可的松的血浆样品。分析物在电喷雾电离的正离子模式下进行检测和定量,使用多重反应监测过渡集,质量电荷(m/z): 21-DOC和17-OHP,皮质醇,可的松和皮质醇-d4 (IS)分别为347.17,311.12,331.17,96.97,363.11,361.18,163.11和367.19,121.24。21-DOC、17-OHP、可的松和皮质醇的浓度与峰面积响应(分析物/IS)在0.25 ~ 50、0.5 ~ 100、1 ~ 200和2 ~ 400 ng/mL范围内呈线性关系。分析物的平均萃取回收率在83% ~ 96%之间。日内和日间变异系数小于13.6%,偏倚范围为-9.2% ~ 12%。皮质醇、可的松、17-OHP测定值分别为21.9 ~ 110、4.33 ~ 12.71、0.37 ~ 1.4 ng/mL。皮质醇-皮质醇比值的测量值在0.08-0.21之间。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Method Development and Clinical Utility for Simultaneous Measurement of 21-Deoxycortisol, 17-Hydroxyprogesterone, Cortisol, and Cortisone Levels in Human Plasma Using UHPLC-MS/MS.

A simple and efficient validated assay for quantifying 21-deoxycortisol (21-DOC), 17-hydroxyprogesterone (17-OHP), cortisol, and cortisone in human plasma has been developed using ultra-high performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS). Analysis of plasma samples were performed on Atlantis dC18 (3 μm) column using a mobile phase of 20.0 mM ammonium acetate and acetonitrile (50:50, v : v) that was delivered at isocratic flow rate 0.3 mL/minute. After addition of d4-cortisol as an internal standard (IS), plasma samples containing 21-DOC, 17-OHP, cortisol, and cortisone were extracted with mixture of dichloromethane and tert-butylmethyl ether 1:2 (v/v). Analytes were detected and quantified in the positive ion mode of electrospray ionization using multiple reaction monitoring transition set at mass to charge (m/z): 347.17 ⟶ 311.12, 331.17 ⟶ 96.97, 363.11 ⟶ 121.00, 361.18 ⟶ 163.11, and 367.19 ⟶ 121.24 for 21-DOC and 17-OHP, cortisol, cortisone, and cortisol-d4 (IS), respectively. The relationship between concentration and peak area response (analyte/IS) were linear over the range of 0.25-50, 0.5-100, 1-200, and 2-400 ng/mL for 21-DOC, 17-OHP, cortisone, and cortisol, respectively. The mean extraction recovery of the analytes was in the range of 83%-96%. The coefficient of variation within and between days was less than 13.6%, and the bias ranged from -9.2% to 12%. The measured level of cortisol, cortisone, and 17-OHP was in the range of 21.9-110, 4.33-12.71, and 0.37-1.4 ng/mL, respectively. Furthermore, the measured value of cortisone-cortisol ratio was in the range of 0.08-0.21.

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来源期刊
CiteScore
4.30
自引率
3.60%
发文量
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审稿时长
17 weeks
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