Li Bingchu, Liu Zhenni, Qin Nijiao, Li Binghui, Chen Zhiyang, Li Ziyang, Lyu Xing, Tan Li, Guo Zhe, Wu Weimin, Xiang Zhongyuan, Liu Zhendong, Long Qichen, Hu Min
Department of Laboratory Medicine, The Second Xiangya Hospital, Central South University, Changsha, Hunan, China; Center for Clinical Molecular Diagnostics, The Second Xiangya Hospital, Central South University, Changsha, Hunan, China.
Department of Laboratory Medicine, The Second Xiangya Hospital, Central South University, Changsha, Hunan, China; Center for Clinical Molecular Diagnostics, The Second Xiangya Hospital, Central South University, Changsha, Hunan, China; National Clinical Research Center for Metabolic Diseases, Department of Metabolism and Endocrinology, The Second Xiangya Hospital of Central South University, Changsha, China.
Talanta. 2025 Dec 1;295:128268. doi: 10.1016/j.talanta.2025.128268. Epub 2025 May 6.
Angiotensin I and II (Ang I, Ang II), aldosterone (ALD), 18-hydroxycorticosterone (18-OHB), 18-hydroxycortisol (18-OHF), and 18-oxocortisol (18-OXOF) are critical biomarkers for the screening, confirming, and subtyping of primary aldosteronism (PA). Liquid chromatography-tandem mass spectrometry (LC-MS/MS) enables simultaneous quantification of multiple analytes; however, obstacles such as labor-intensive manual procedures, complex biological matrices, structural analog interference, and the physicochemical diversity of target analytes limit its application and promotion in clinical laboratories. This study established and validated a robust LC-MS/MS method based on automated magnetic-bead-assisted sequential extraction (MBASE) for the simultaneous detection and quantitation of Ang I, Ang II, ALD, 18-OHB, 18-OHF, and 18-OXOF. The method achieved complete chromatographic resolution with no interference from endogenous hormones or exogenous medications. The analytical recoveries ranged from 86.14 % to 112.89 %, with total imprecisions between 2.93 % and 12.89 %. The MBASE demonstrated a strong correlation with traditional SPE for all six analytes (R: 0.967-0.997), with biases ranging from -4.0 % to 5.1 %. Compared to traditional SPE process, the MBASE workflow reduced total processing time by 46 % and manual handling time by 77 %. The clinical application of this LC-MS/MS method indicated that Ang I and Ang II are effective screening biomarkers, and ALD, 18-OHB, 18-OHF, and 18-OXOF could be used to differentiate PA subtypes. Overall, the newly developed MBASE-LC-MS/MS method exhibits excellent performance and significant advantages for PA diagnosis. It can be a promising tool in the early screening and subtyping for PA and enhance automated mass spectrometry processes in clinical application.
血管紧张素I和II（Ang I、Ang II）、醛固酮（ALD）、18-羟皮质酮（18-OHB）、18-羟皮质醇（18-OHF）和18-氧代皮质醇（18-OXOF）是原发性醛固酮增多症（PA）筛查、确诊及分型的关键生物标志物。液相色谱-串联质谱法（LC-MS/MS）能够同时对多种分析物进行定量；然而，诸如劳动强度大的手工操作、复杂的生物基质、结构类似物干扰以及目标分析物的物理化学多样性等障碍限制了其在临床实验室中的应用和推广。本研究建立并验证了一种基于自动磁珠辅助顺序萃取（MBASE）的稳健LC-MS/MS方法，用于同时检测和定量Ang I、Ang II、ALD、18-OHB、18-OHF和18-OXOF。该方法实现了完全的色谱分离，不受内源性激素或外源性药物的干扰。分析回收率在86.14%至112.89%之间，总不精密度在2.93%至12.89%之间。MBASE与传统固相萃取法对所有六种分析物均显示出强相关性（R：0.967 - 0.997），偏差范围为-4.0%至5.1%。与传统固相萃取过程相比，MBASE工作流程将总处理时间减少了46%，手工操作时间减少了77%。这种LC-MS/MS方法的临床应用表明，Ang I和Ang II是有效的筛查生物标志物，而ALD、18-OHB、18-OHF和18-OXOF可用于区分PA亚型。总体而言，新开发的MBASE-LC-MS/MS方法在PA诊断方面表现出优异的性能和显著优势。它可能成为PA早期筛查和分型的有前途的工具，并在临床应用中增强自动质谱分析过程。
