Liu Zhenni, Jin Lizi, Zhang Jiangtao, Zhang Tianjiao, Zeng Jie, Zhou Weiyan, Zhang Chuanbao
National Center for Clinical Laboratories, Institute of Geriatric Medicine, Chinese Academy of Medical Sciences, Beijing Hospital/National Center of Gerontology, P. R. China.
Chinese Academy of Medical Sciences and Peking Union Medical College, No. 1 Dahua Road, Dongcheng District, Beijing 100730, P. R. China.
Anal Methods. 2023 Jan 26;15(4):492-501. doi: 10.1039/d2ay01646j.
Plasma renin activity (PRA) is recommended as the first screening indicator for primary aldosteronism. Immunoassays and liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods have been developed for quantifying PRA, but the interchangeability across assays and laboratories was suboptimal, which predominantly related to the differences in the plasma incubation strategy. This study aims to establish and validate a designed comparison method based on LC-MS/MS. The sensitivity, matrix effect, precision, accuracy, and storage stability were validated according to the Clinical Laboratory Standard Institution (CLSI) C-62A guidelines. The plasma incubation procedure was optimized to achieve maximum PRA results. The short-term stability of PRA plasma was assessed at 4 °C and room temperature (RT) for specific time points. Differences from the baseline were calculated using a one-way analysis of variance. The designed comparison method for PRA measurement exhibits excellent performance characteristics. The results from the 2022 national external quality assessment scheme for PRA showed good consistency of the developed method with other LC-MS/MS methods (relative biases: -6.8% to 4.6%), which demonstrated the reliability of the established method. Two sets of generation buffers were optimized to maximize the renin activity. The acetate buffer was recommended to be used in laboratory practice due to better metrological sensitivity. PRA plasma is stable for one day at 4 °C and RT. In summary, a reliable, traceable, and reproducible LC-MS/MS method for determining PRA was well-established and validated. The recommended incubation protocol is hoped to reduce the discrepancy in Ang1 generation. The evaluated short-term stability for PRA plasma could provide flexibility in clinical practice.
血浆肾素活性（PRA）被推荐作为原发性醛固酮增多症的首个筛查指标。已经开发了免疫测定法和液相色谱-串联质谱法（LC-MS/MS）来定量PRA，但各检测方法和实验室之间的互换性欠佳，这主要与血浆孵育策略的差异有关。本研究旨在建立并验证一种基于LC-MS/MS的设计比较方法。根据临床实验室标准协会（CLSI）C-62A指南对灵敏度、基质效应、精密度、准确度和储存稳定性进行了验证。优化了血浆孵育程序以获得最大的PRA结果。在4℃和室温（RT）下的特定时间点评估PRA血浆的短期稳定性。使用单向方差分析计算与基线的差异。所设计的PRA测量比较方法具有优异的性能特征。2022年PRA全国外部质量评估计划的结果表明，所开发的方法与其他LC-MS/MS方法具有良好的一致性（相对偏差：-6.8%至4.6%），这证明了所建立方法的可靠性。优化了两组生成缓冲液以最大化肾素活性。由于计量灵敏度更高，建议在实验室实践中使用乙酸盐缓冲液。PRA血浆在4℃和室温下可稳定一天。总之，建立并验证了一种可靠、可溯源且可重复的用于测定PRA的LC-MS/MS方法。所推荐的孵育方案有望减少血管紧张素1生成的差异。所评估的PRA血浆短期稳定性可为临床实践提供灵活性。
