College of Chemistry, Beijing University of Chemical Technology, Beijing 100029, PR China.
KingMed Diagnostics (Beijing) Co., Ltd., Beijing 100103, PR China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2024 Feb 15;1234:124012. doi: 10.1016/j.jchromb.2024.124012. Epub 2024 Jan 14.
Aldosterone (ALD), its precursor 18-hydroxycorticosterone (18-OHB) and its metabolite tetrahydroaldosterone (TH-ALD) are important biomarkers for the diagnosis of primary aldosteronism (PA). Liquid chromatography-tandem mass spectrometry (LC-MS/MS) is increasingly utilized in the detection of small molecules of hormones because it has advantages in terms of specificity and sensitivity. The objective of this study is to develop a new LC-MS/MS method for the simultaneous quantification of ALD (free), 18-OHB, and TH-ALD in human urine and attempt to diagnose primary aldosteronism using different indicators. The urine samples were treated with a solid-phase extraction pretreatment technique and the three analytes were separated on a reversed-phase column and detected on a triple quadrupole mass spectrometer. The established method was validated according to CLSI C62-A standard guidelines. The calibration ranges from 25 pg/mL to 5000 pg/mL for aldosterone (free), 18-hydroxycorticosterone and tetrahydroaldosterone, and the lower limit of quantification for these three analytes was 25 pg/mL. The matrix effects and recoveries of these three analytes ranged from 85.1 % to 115 % and from 86.3 % to 114 %, respectively. The intra-day and inter-day precision ranged from 1.29 % to 6.78 % and from 1.77 % to 8.64 %, respectively. The performance of the method met the requirements of the guidelines. 40 clinical urine samples including 22 PA patients and 18 non-PA patients were detected, and the ROC curves of three diagnostic indicators were established. The area under the curve (AUC) of ALD (free) is the biggest, so ALD (free) was the best compound to be used as a diagnostic indicator in this study. When the cut-off point was taken as 141 ng/24-h, the sensitivity was 72.7 % and the specificity was 88.9 %. We developed and validated an LC-MS/MS method for the simultaneous quantification of ALD (free), 18-OHB and TH-ALD in human urine. Our study provides a reference for the use of new biomarkers for the diagnosis of primary aldosteronism.
醛固酮 (ALD)、其前体 18-羟皮质酮 (18-OHB) 和其代谢物四氢醛固酮 (TH-ALD) 是原发性醛固酮增多症 (PA) 诊断的重要生物标志物。液相色谱-串联质谱 (LC-MS/MS) 因其在特异性和灵敏度方面的优势，越来越多地用于检测激素的小分子。本研究旨在建立一种新的 LC-MS/MS 方法，用于同时定量测定人尿液中的 ALD(游离)、18-OHB 和 TH-ALD，并尝试使用不同的指标来诊断原发性醛固酮增多症。尿液样本采用固相萃取预处理技术，三种分析物在反相柱上分离，在三重四极杆质谱仪上检测。该方法按照 CLSI C62-A 标准指南进行验证。ALd(游离)、18-羟基皮质酮和四氢醛固酮的校准范围为 25 pg/mL 至 5000 pg/mL，这三种分析物的定量下限均为 25 pg/mL。这三种分析物的基质效应和回收率分别在 85.1%至 115%和 86.3%至 114%之间。日内和日间精密度分别在 1.29%至 6.78%和 1.77%至 8.64%之间。方法性能符合指南要求。检测了 40 例临床尿液样本，包括 22 例 PA 患者和 18 例非 PA 患者，建立了三种诊断指标的 ROC 曲线。ALd(游离)的曲线下面积 (AUC)最大，因此 ALd(游离)是本研究中作为诊断指标的最佳化合物。当以 141 ng/24-h 为截断值时，灵敏度为 72.7%，特异性为 88.9%。我们建立并验证了一种 LC-MS/MS 方法，用于同时定量测定人尿液中的 ALD(游离)、18-OHB 和 TH-ALD。本研究为使用新的生物标志物诊断原发性醛固酮增多症提供了参考。
