Xue Xue, Chen Hucheng, Xia Qingqing, He Ping, Lv Yuan, Liu Peng, Yang Yan, Wu Chaochao, Gao Qiang, Peng Jun, Yu Yang, Wang Feng, Qu Wei, Wang Zizheng
Department of Nuclear Medicine, Nanjing First Hospital, Nanjing Medical University, Nanjing, Jiangsu, China.
Central Laboratory, First People's Hospital of Taizhou, Taizhou, Zhejiang, China.
Ann Clin Lab Sci. 2019 Mar;49(2):204-211.
The measurement of plasma catecholamines (CAs) including dopamine (DA), epinephrine (E), and norepinephrine (NE) and their derivatives including metanephrine (MN), normetanephrine (NMN), vanillylmandelic acid (VMA), and homovanillic acid (HVA) has been used in the diagnosis of pheochromocytoma and paraganglioma (PPGL) and primary hypertension (PH) but are typically detected individually when clinical testing. In this study, pre-column derivatization with dansyl chloride (DNS-Cl) combined with an ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed to simultaneously quantify HVA, VMA, MN, NMN, DA, E, and NE in the plasma from patients with PPGL and PH.
Plasma samples were extracted by acetonitrile and derivatized with DNS-Cl, followed by reverse phase separation and triple quadruple detection. Quantification of the CAs and their derivatives in 10 PPGL, 10 PH, and 100 healthy subjects was performed by UPLC-MS/MS analysis.
All the values of detected CAs/derivatives were in the linearity ranges of the fitted curves. The expression levels of the seven CAs in the PPGL and PH patients were significantly higher than the healthy controls, suggesting increased CA production in the former. There were significant differences in plasma NE, NMN, and VMA levels between the PPGL and PH patients, but there was no significant difference in plasma E, MN, DA, and HVA. A discriminant analysis showed that 90% of the final cases were classified correctly based on the detected CAs/derivatives.
Our results show that the combined detection of the seven CAs/derivatives could be used for the clinical diagnosis of PPGL and PH.
血浆儿茶酚胺（CAs）包括多巴胺（DA）、肾上腺素（E）和去甲肾上腺素（NE）及其衍生物包括间甲肾上腺素（MN）、去甲间甲肾上腺素（NMN）、香草扁桃酸（VMA）和高香草酸（HVA）的测定已用于嗜铬细胞瘤和副神经节瘤（PPGL）以及原发性高血压（PH）的诊断，但在临床检测时通常是单独检测。在本研究中，开发了一种用丹磺酰氯（DNS-Cl）进行柱前衍生结合超高效液相色谱-串联质谱（UPLC-MS/MS）的方法，以同时定量PPGL和PH患者血浆中的HVA、VMA、MN、NMN、DA、E和NE。
血浆样本用乙腈提取并用DNS-Cl衍生，然后进行反相分离和三重四极杆检测。通过UPLC-MS/MS分析对10例PPGL患者、10例PH患者和100例健康受试者的CAs及其衍生物进行定量。
所有检测到的CAs/衍生物的值均在拟合曲线的线性范围内。PPGL和PH患者中七种CAs的表达水平显著高于健康对照，表明前者的CA生成增加。PPGL和PH患者之间血浆NE、NMN和VMA水平存在显著差异，但血浆E、MN、DA和HVA水平无显著差异。判别分析表明，基于检测到的CAs/衍生物，90%的最终病例被正确分类。
我们的结果表明，联合检测七种CAs/衍生物可用于PPGL和PH的临床诊断。
