Wang Yuting, Li Quan, Deng Yuhang, Wu Wenqing, Zhang Cuiping, Zheng Yichi, Guan Ming, Jiang Haoqin
Department of Laboratory Medicine, Huashan Hospital, Fudan University, Shanghai, 200040, China.
Huashan CSF Laboratory, Huashan Hospital, Fudan University, Shanghai, 200040, China.
Pract Lab Med. 2025 Apr 17;45:e00471. doi: 10.1016/j.plabm.2025.e00471. eCollection 2025 Jul.
Catecholamines (CAs) and their metabolites in human cerebrospinal fluid (CSF) and plasma are potential biomarkers of Alzheimer's disease (AD) and facilitate early diagnosis. Liquid chromatography-tandem mass spectrometry is the gold standard method for analyzing CAs. The objective of this study was to develop and validate a liquid chromatography-tandem mass spectrometry assay capable of simultaneously quantifying dopamine (DA), epinephrine (E), norepinephrine (NE), metanephrine (MN), normetanephrine (NMN), and 3-methoxytyramine (3-MT) in both human CSF and plasma. Samples were processed by solid-phase extraction with a weak cation exchange adsorbent and then separated using an ultra-performance reversed-phase chromatography column. Analyte detection was performed using a triple quadrupole mass spectrometer operated in positive-ion multiple reaction monitoring mode. The developed assay was validated according to standard guidelines. The linearity, specificity, precision, accuracy, carryover and stability were assessed to ensure compliance with specified criteria. The lower limits of quantification for DA, E, NE, MN, NMN, and 3-MT were 4.5, 2.5, 4.5, 2.5, 2, and 0.3 pg mL, respectively. The total runtime for a single sample was 6.5 min. These results demonstrated that the method was sensitive, rapid, and reliable for the simultaneous quantification of DA, E, NE, MN, NMN, and 3-MT in clinical practice. We successfully detected CAs and their metabolites in plasma and CSF samples from patients with normal cognition and AD. This study demonstrates an efficient laboratory workflow for high-throughput analysis of CAs and their metabolites and lays a foundation for further studies on AD biomarkers.
儿茶酚胺（CAs）及其在人脑脊液（CSF）和血浆中的代谢产物是阿尔茨海默病（AD）的潜在生物标志物，有助于早期诊断。液相色谱 - 串联质谱法是分析CAs的金标准方法。本研究的目的是开发并验证一种液相色谱 - 串联质谱测定法，该方法能够同时定量测定人CSF和血浆中的多巴胺（DA）、肾上腺素（E）、去甲肾上腺素（NE）、间甲肾上腺素（MN）、去甲间甲肾上腺素（NMN）和3 - 甲氧基酪胺（3 - MT）。样品通过用弱阳离子交换吸附剂进行固相萃取处理，然后使用超高效反相色谱柱进行分离。使用在正离子多反应监测模式下运行的三重四极杆质谱仪进行分析物检测。根据标准指南对所开发的测定法进行验证。评估线性、特异性、精密度、准确度、残留和稳定性以确保符合指定标准。DA、E、NE、MN、NMN和3 - MT的定量下限分别为4.5、2.5、4.5、2.5、2和0.3 pg/mL。单个样品的总运行时间为6.5分钟。这些结果表明该方法在临床实践中对同时定量测定DA、E、NE、MN、NMN和3 - MT是灵敏、快速且可靠的。我们成功检测了认知正常患者和AD患者血浆和CSF样品中的CAs及其代谢产物。本研究展示了一种用于CAs及其代谢产物高通量分析的高效实验室工作流程，并为进一步研究AD生物标志物奠定了基础。
