利用毛细管电泳电化学发光(CE-ECL)法检测了尿样中的香草扁桃酸(VMA)和高香草酸(HVA),得到了最佳的分离检测条件:50 mmol/L磷酸盐缓冲溶液(pH 8.2);分离电压16 kV;检测电势1.2 V(相对于饱和甘汞电极).在最佳条件下,VMA和HVA的浓度检测限(S/N=3)分别为5.0×10-7 mol/L和6.1×10-7 mol/L,线性回归系数分别为0.999 3和0.997 9.并以此方法将尿液中的VMA和HVA在10 min内有效地分离检测,不受其他干扰,得到加标回收率分别为98%和99%,取得了满意的结果.
Vanilla mandelic acid(VMA)and homovanillic acid(HVA)in the urine were detected by capillary electrophoresis with electrochemiluminescence detection(CE-ECL).The following conditions were suitable for the determination of VMA and HVA:running buffer,50 mmol/L NaH2PO4-Na2HPO4(pH=8.2);separation voltage,16 kV;detection potential,1.2 V(vs.saturated calomel electrode(SCE)).Under the optimum conditions of detection,satisfactory sensitivities(LOD of 5.0×10-7 mol/L for VMA and 6.1×10-7 mol/L for HVA at S/N=3)were obtained,the correlation coefficients of the method were 0.999 3 and 0.997 9,respectively.The determination of these metabolites in human urine was completed within 10 min without any interferences,the recoveries were VMA 98% and HVA 99%,the result obtained is satisfactory.
[1] SADILKOVA K,DUGAW K,BENJAMIN D,et al.Analysis of vanillylmandelic acid and homovanillic acid by UPLC–MS/MS in serum for diagnostic testing for neuroblastoma[J].Clinica Chimica Acta,2013,424:253-257.DOI:10.1016/j.cca.2013.06.024.
[2] 许育丹.重氮法和微柱比色法检测尿液香草扁桃酸的方法比较[J].实用医技杂志,2005,12(03A):578-579.
[3] 陈曼娜,廖瑛.微柱比色法检测尿液VMA含量的评价[J].现代检验医学杂志,2003,18(1):14-15. CHEN Manna,LIAO Ying.To envalue the way of using micro-column chromatography determine VMA in the urine[J].Journal Of Modern Laboratory Medicine,2003,18(1):14-15.
[4] TOKUDA T,TOKIEDA T,ANAZAWA A,et al.Simultaneous determination of vanillylmandelic acid,homovanillic acid and creatinine in urine by reversed-phase ion-pair chromatography[J].Journal of Chromatography B:Biomedical Sciences And Applications,1990,530:418-423.DOI:10.1016/s0378-4347(00)82344-0.
[5] 张华,李永兴.高效液相电化学法检测尿儿茶酚胺和VMA诊断嗜铬细胞瘤[J].现代检验医学杂志,2003,18(3):8-9. ZHANG Hua,LI Yongxing.Urinary catecholamines and VMA measured by high performance liquid chromatography(HPLC)-electrochemical detector(ECD)for diagnosis of pheochromocytoma[J].Journal of Modern Laboratory Medicine,2003,18(3):8-9.
[6] DENG Biyang,SU Caina,KANG Yanhui.Determination of norfloxacin in human urine by capillary electrophoresis with electrochemiluminescence detection[J].Analytical And Bioanalytical Chemistry,2006,385(7):1336-1341.DOI:10.1007/s00216-006.
[7] AN Dong,CHEN Zhuqiu,ZHENG Jiachun,et al.Determination of biogenic amines in oysters by capillary electrophoresis coupled with electrochemiluminescence[J].Food Chemistry,2015,168:1-6.DOI:10.1016/j.foodchem.2014.07.019.
[8] LI Jianguo,YAN Qinyi,GAO Yunlong,et al.Electrogenerated chemiluminescence detection of amino acids based on precolumn derivatization coupled with capillary electrophoresis separation[J].Analytical Chemistry,2006,78(8):2694-2699.DOI:10.1021/ac052092m.
[9] 高萍,徐佳,孙玉倩,等.毛细管电泳检测尿儿茶酚胺代谢产物及在诊断嗜铬细胞瘤中的应用[J].哈尔滨医科大学学报,2008,42(5):511. GAO Ping,XU Jia,SUN Yuqian,et al.Determination of urine catecholamine metabolic products by capillary electrophoresis for diagnosis of pheochromocytoma[J].Journal Of Harbin Medical University,2008,42(5):511.
[10] JIN Wenrui,WENG Qianfeng,WU Jianren.Determination of bovine serum albumin by capillary zone electrophoresis with end-column amperometric detection at the carbon fiber microdisk array electrode[J].Analytica Chimica Acta,1997,342(1):67-74.DOI:10.1016/s0003-2670(96)00530-2.
[11] JIN Wenrui,YU Daiqing,DONG Qian,et al.A new method of quantification of pipemidic acid by capillary zone electrophoresis with end-column amperometric detection[J].Electrophoresis,2000,21(5):925-929.
