库仑阵列电化学测定尿儿茶酚胺代谢物方法的建立及其在神经母细胞瘤诊断中的应用  

Determination of urinary catecholamines metabolites by coulometric electrode array system （CoulArray） in diagnosis of neuroblastoma

作　　者：常会波[1] 保睿[1] 叶辉[2] 李龙[2] 张晓伦[2] 邹继珍[3] 刘卓[1] 吴建新[1] 

机构地区：[1]首都儿科研究所生物化学研究室,北京100020 [2]首都儿科研究所外科,北京100020 [3]首都儿科研究所病理科,北京100020

出　　处：《中华小儿外科杂志》2010年第8期570-574,共5页Chinese Journal of Pediatric Surgery

基　　金：国家自然科学基金项目（30671156）北京市自然科学基金（7042017,5072014）北京市优秀人才课题资助项目（20081D0303200106）

摘　　要：目的 建立以反相液相色谱-库仑阵列电化学联用紫外检测法同时测定随机尿液中儿茶酚胺代谢产物方法,并应用于0～10岁儿童正常值范围的确立及神经母细胞瘤的诊断.方法 反相液相色谱梯度洗脱法分离样品,库仑阵列电化学定量检测VMA和HVA的浓度,紫外检测器检测Cr的浓度.以VMA/Cr和HVA/Cr表示尿儿茶酚胺代谢物的含量.对该法进行系统性评价后应用于120例健康儿童和15例神经母细胞瘤患儿随机尿样的测定中.结果 VMA、HVA和Cr的日内变异系数和日问变异系数均小于5%,线性相关系数均大于0.9990.回收率均大于95%.健康儿童VMA/Cr（2.99±1.77）μg/mg和HVA/Cr（3.40±1.50）μg/mg与年龄有显著相关性（P＜0.001）,0～2岁组[VMA/Cr（4.04±1.24）μg/mg,HVA/Cr（4.87±1.81）μg/mg]与其他年龄组[2～4岁：VMA/Cr（3.02±0.81）μg/mg,HVA/Cr（3.46±1.06）μg/mg;4～6岁：VMA/Cr（2.68±0.95）μg/mg,HVA/Cr（2.75±1.04）μg/mg;6～10岁：VMA/Cr（2.32±0.95）μg/mg,HVA/Cr（2.61±0.84）μg/mg]差异均有统计学意义（P＜0.001）.以低于（-x）＋3s作为正常值,超过此范围诊断为神经母细胞瘤,对患儿诊断的符合率达93%.结论 本研究建立了同时快速准确检测随机尿样VMA、HVA和Cr的方法,并分年龄组建立了0～10岁儿童的正常值范围,为神经母细胞瘤的诊断和筛查提供了有效的参考指标和简单易行、准确灵敏的生化检验手段,也可用于与儿茶酚胺异常分泌相关的其他研究领域.Objective To establish reverse phase liquid chromatographic（RPLC） with CoulArray（coulometric electrode array system） in assaying urinary catecholamines metabolites and establish normal reference ranges for neuroblastoma diagnosis. Methods Samples were processed by binary gradient reversed phase chromatography. Vanillylmandelic acid （VMA） and homovanillic acid（HVA）were assayed by CoulArray, and creatinine（Cr）was by UV detector. The contents of catecholamines metabolites were expressed as VMA/Cr and HVA/Cr. After calibration, this method was extended to random urine samples from healthy children （n = 120） and neuroblastoma patients （n = 15）. Results The results showed good correlation （r＞0.9990） with VMA, HVA and Cr. The methodological recovery was more than 95%, and the intra- and inter-day coefficients of variation were both less than 5%. VMA/Cr（2.99 ± 1.77 μg/mg） and HVA/Cr（3.40±1.50μg/mg）decreased as the age increased （P＜0. 001 ） in normal children, and there were significant differences between 0～ 2 years group （VMA/Cr 4. 04 ± 1.24μg/mg, HVA/Cr 4.87 ± 1.81μg/mg） and the old children groups（2～4 years：VMA/Cr 3.02 ± 0.81μg/mg, HVA/Cr 3.46 ± 1.06 μg/mg; 4～6 years： VMA/Cr 2.68 ± 0.95 μg/mg, HVA/Cr 2. 75 ± 1.04 μg/mg; 6～10 years： VMA/Cr 2.32±0.95μg/mg, HVA/Cr 2.61±0.84μg/mg） （P＜0. 001 ）. The normal range was less than （-x） ＋ 3s; values above it were suggestive of neuroblastoma. The diagnostic accuracy of the CoulArray was 93%. Conclusions This method was proven to be simple, accurate, highly sensitive,and could be applied simultaneously to the analysis of random urinary VMA, HVA and Cr. The normal VMA/Cr and HVA/Cr variation of different age groups have been established. This will be useful for the diagnosis and monitoring of neuroblastoma.

关 键 词：电化学 色谱法 液相 高香草酸 香草扁桃酸 神经母细胞瘤 

分 类 号：R739.4[医药卫生—肿瘤]