神经母细胞瘤患儿尿中儿茶酚胺含量的HPLC测定

Determination of urinary catecholamines with neuroblastoma in children by HPLC

目的建立高效液相色谱法（HPLC）测定尿中去甲肾上腺素（NE）、肾上腺素（E）和多巴胺（DA）三种儿茶酚胺类物质的方法,为临床确诊神经母细胞瘤（neuroblastoma,NB）提供诊断手段.方法利用高效液相色谱及荧光检测器,采用Zorbax SB-Aq-C18（4.6mm×150mm,5μm）色谱柱,使用0.02mol/L KH2PO4溶液为流动相,对正常儿童和神经母细胞瘤患儿24h尿中NE、E和DA进行测定.结果NE、E和DA的日内精密度分别为5.46%、3.96%、4.49%,日间精密度分别为6.48%、4.67%、5.43%.NE、E和DA低、中、高3种浓度加标回收率分别为92.20%～94.38%、94.60%～95.85%、88.43%～91.68%.NE、E的线性范围为25～800 μg/L、DA的为50～1600μg/L.最低检测限分别为2.5、1.5、0.5μg/L.结论建立的方法精密度好、准确度高、特异性强,适用于临床神经母细胞瘤的诊断.

Objective To develop the method of high performance liquid chromatography （HPLC） to determine the level of catecholamines（CAs） in 24h urinary,such as norepinephrine（NE） ,epinephrine （E） ,dopamine（DA） ,and to apply in the diagnosis of neuroblastoma（NB）. Methods The determination was performed in a Zorbax SB-Aq-C18 （4.6 × 150mm,Sμm） column with a mobile phase of 0.02mol/L KH2PO4 solution, and with fluorescence detector. In the condition,the levels of 24h urinary NE, E, DA were detected by HPLC in healthy children, and in children with NB. Results The recovery rates of NE, E and DA were 92.20% - 94.38% ,94.60% - 95.85% and 88.43% - 91.68%, respectively, the corresponding with-day precisions （n = 6） were 5.46%, 3.96% and 4.49%, respectively, and the between-day precisions （ n = 6） were 6.48 %, 4.67 % and 5.43 %, respectively. The linear scopes of NE and E were 25 - 800p.g/L, respectively, the linear scope of DA was 50 - 1600μg/L. The least detections were 2.5, 1.5,0.5μg/L, respectively. Conclusion This method is simple,accurate and high specificity,and can be used for the clinical diagnosis of NB.