采用磁固相萃取与液相色谱-串联质谱联用技术同时定量检测继发性高血压的生物标志物谱。

A biomarker panel of secondary hypertension is simultaneously quantified by coupling of magnetic solid-phase extraction and liquid chromatography-tandem mass spectrometry.

机构信息

College of Life Sciences, University of Chinese Academy of Sciences, Beijing, China.

BGI Genomics, Shenzhen, China.

出版信息

Rapid Commun Mass Spectrom. 2024 Mar 30;38(6):e9703. doi: 10.1002/rcm.9703.

DOI:10.1002/rcm.9703

PMID:38356091

Abstract

RATIONALE

Secondary hypertension is often caused by activation of complex multi-organ endocrine systems, while renin activity indicated by angiotensins (Angs), aldosterone (ALD) and cortisol (COR) in such systems are generally accepted as its diagnostic markers. As antibody-based methods cannot offer comparable quantification for these biomarkers, a liquid chromatography (LC)-tandem mass spectrometry (MS/MS)-based approach was developed to quantify them simultaneously and accurately.

METHODS

Five different beads for magnetic solid-phase extraction (MSPE) were evaluated towards their enrichment efficiency for these biomarkers. An LC system with optimized elution gradient and a triple-quadrupole MS with tuned parameters were coupled to quantitatively monitor the extracted analytes. The method performance was further examined such as linearity, precision, stability, recovery rate and matrix effect. Based on the developed method, the abundance of Ang II, ALD and COR in plasma was measured and the quantification was compared with that derived from commercial ELISA kits.

RESULTS

As compared with other MSPEs, Angs, ALD and COR were highly enriched by the HLB magnetic beads with satisfactory recoveries. These analytes were simultaneously quantified by LC/MS/MS and all the method parameters for quantification were well matched with the requirements of clinical testing. Comparison of the quantitative results derived from ELISA and LC/MS/MS exhibited that the two methods offered basically comparable values with Pearson r values at 0.896, 0.895 and 0.835, respectively. The stability test for plasma Angs at room temperature indicated that the abundance of Ang II was relatively stable within 3 h, whereas that of Ang I and Ang 1-7 was time-dependently changed.

CONCLUSIONS

Coupling of HLB beads and LC/MS/MS thus enables simultaneous quantification of a set of biomarkers related to secondary hypertension.

摘要

原理

继发性高血压通常由复杂的多器官内分泌系统激活引起，而该系统中血管紧张素（Angs）、醛固酮（ALD）和皮质醇（COR）等物质所指示的肾素活性通常被认为是其诊断标志物。由于基于抗体的方法无法对这些生物标志物进行可比的定量，因此开发了一种基于液相色谱（LC）-串联质谱（MS/MS）的方法来同时准确地对其进行定量。

方法

评估了五种不同的磁固相萃取（MSPE）珠对这些生物标志物的富集效率。优化了洗脱梯度的 LC 系统和调谐参数的三重四极杆 MS 被耦合起来，以定量监测提取的分析物。进一步检查了方法性能，如线性、精密度、稳定性、回收率和基质效应。基于所开发的方法，测量了血浆中 Ang II、ALD 和 COR 的丰度，并将其定量与商业 ELISA 试剂盒得出的结果进行了比较。

结果

与其他 MSPE 相比，HLB 磁珠高度富集了 Angs、ALD 和 COR，且回收率令人满意。这些分析物通过 LC/MS/MS 同时进行定量，所有定量参数都与临床检测要求相匹配。ELISA 和 LC/MS/MS 定量结果的比较表明，两种方法基本提供了可比的值，皮尔逊 r 值分别为 0.896、0.895 和 0.835。室温下血浆 Angs 的稳定性测试表明，Ang II 的丰度在 3 小时内相对稳定，而 Ang I 和 Ang 1-7 的丰度则随时间而变化。