Chan E C, Ho P C
Department of Pharmacy, National University of Singapore, Singapore.
Rapid Commun Mass Spectrom. 2000;14(21):1959-64. doi: 10.1002/1097-0231(20001115)14:21<1959::AID-RCM117>3.0.CO;2-T.
An assay of norepinephrine (NE), epinephrine (E), dopamine (DA), normetanephrine (NE) and metanephrine (MN) based on high-performance liquid chromatography (HPLC) in combination with atmospheric pressure chemical ionization mass spectrometry (APcI-MS) is described. The catecholamines and metanephrines were extracted from urine and aqueous samples using Bio-Rex 70 cation-exchange resin and subjected to analysis by HPLC/APcI-MS. The separation was performed on a C18 column in 50 mM ammonium formate buffer, pH 3.0, using a flow rate of 0.8 mL/min. Acetonitrile was added post-column at a flow rate of 0.2 mL/min via a post-column addition tee. The total analysis time was 6.5 min. The quantitative analysis was performed using 3,4-dihydroxybenzylamine (DHBA) as the internal standard (I.S.). Selected ion monitoring detection was applied: m/z 170 (for NE), 184 (for E and NM), 154 (for DA), 198 (for MN) and 140 (for DHBA, I.S.). The limits of quantitation were 5 ng/mL for NE, E and DA and 2.5 ng/mL for NM and MN. The recovery ranged from 75 to 83% for each analyte. The method was found to be simple and highly selective for the determination of catecholamines and metanephrines in the urine of patients suspected of pheochromocytoma.
本文描述了一种基于高效液相色谱（HPLC）结合大气压化学电离质谱（APcI-MS）的去甲肾上腺素（NE）、肾上腺素（E）、多巴胺（DA）、去甲变肾上腺素（NE）和变肾上腺素（MN）的检测方法。儿茶酚胺类和变肾上腺素类物质通过Bio-Rex 70阳离子交换树脂从尿液和水性样品中提取出来，并采用HPLC/APcI-MS进行分析。分离在C18柱上进行，流动相为pH 3.0的50 mM甲酸铵缓冲液，流速为0.8 mL/min。乙腈通过柱后添加三通以0.2 mL/min的流速在柱后添加。总分析时间为6.5分钟。定量分析使用3,4-二羟基苄胺（DHBA）作为内标（I.S.）。采用选择离子监测检测：m/z 170（用于NE）、184（用于E和NM）、154（用于DA）、198（用于MN）和140（用于DHBA，内标）。NE、E和DA的定量限为5 ng/mL，NM和MN的定量限为2.5 ng/mL。每种分析物的回收率在75%至83%之间。该方法被发现对于疑似嗜铬细胞瘤患者尿液中儿茶酚胺类和变肾上腺素类物质的测定简单且具有高度选择性。
