CAJ | 학술논문

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.
목적 건립이반상액상색보-고륜진렬전화학련용자외검측법동시측정수궤뇨액중인다분알대사산물방법,병응용우0～10세인동정상치범위적학립급신경모세포류적진단.방법 반상액상색보제도세탈법분리양품,고륜진렬전화학정량검측VMA화HVA적농도,자외검측기검측Cr적농도.이VMA/Cr화HVA/Cr표시뇨인다분알대사물적함량.대해법진행계통성평개후응용우120례건강인동화15례신경모세포류환인수궤뇨양적측정중.결과 VMA、HVA화Cr적일내변이계수화일문변이계수균소우5%,선성상관계수균대우0.9990.회수솔균대우95%.건강인동VMA/Cr(2.99±1.77)μg/mg화HVA/Cr(3.40±1.50)μg/mg여년령유현저상관성(P＜0.001),0～2세조[VMA/Cr(4.04±1.24)μg/mg,HVA/Cr(4.87±1.81)μg/mg]여기타년령조[2～4세:VMA/Cr(3.02±0.81)μg/mg,HVA/Cr(3.46±1.06)μg/mg;4～6세:VMA/Cr(2.68±0.95)μg/mg,HVA/Cr(2.75±1.04)μg/mg;6～10세:VMA/Cr(2.32±0.95)μg/mg,HVA/Cr(2.61±0.84)μg/mg]차이균유통계학의의(P＜0.001).이저우(-x)+3s작위정상치,초과차범위진단위신경모세포류,대환인진단적부합솔체93%.결론 본연구건립료동시쾌속준학검측수궤뇨양VMA、HVA화Cr적방법,병분년령조건립료0～10세인동적정상치범위,위신경모세포류적진단화사사제공료유효적삼고지표화간단역행、준학령민적생화검험수단,야가용우여인다분알이상분비상관적기타연구영역.
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.