高等学校化学学报 ›› 2013, Vol. 34 ›› Issue (12): 2699.doi: 10.7503/cjcu20130835

• 分析化学 • 上一篇    下一篇

神经递质类相关物质的毛细管电泳分离及实际尿液的检测应用

杨娇美1, 任杰2, 刘慧青1, 王永乐1, 徐中其1   

  1. 1. 东华大学化学化工与生物工程学院, 上海 201620;
    2. 上海体育学院运动科学学院, 上海200438
  • 收稿日期:2013-08-29 出版日期:2013-12-10 发布日期:2013-11-01
  • 作者简介:任 杰,男,博士,副教授,主要从事体育运动科学研究. E-mail:renjie@sus.edu.cn;徐中其,男,博士,副教授,主要从事毛细管电泳分离技术研究. E-mail:chemxzq@dhu.edu.cn
  • 基金资助:

    上海市科委“创新行动计划”部分地方院校能力建设专项(批准号:10490503600)和中央高校基本科研业务费专项(批准号:2012D10514)资助.

Separation and Detection of Neurotransmitter-related Substances in Urine Sample by Capillary Electrophoresis

YANG Jiao-Mei1, REN Jie2, LIU Hui-Qing1, WANG Yong-Le1, XU Zhong-Qi1   

  1. 1. College of Chemistry, Chemical Engineering and Biotechnology, Donghua University, Shanghai 201620, China;
    2. School of Kinesiology, Shanghai University of Sport, Shanghai 200438, China
  • Received:2013-08-29 Online:2013-12-10 Published:2013-11-01

摘要:

建立了单胺类神经递质(5-羟色胺、多巴胺和肾上腺素)、神经递质类代谢产物(高香草酸、5-羟吲哚乙酸和香草扁桃酸)及神经递质类前体(精氨酸和酪氨酸)混合物的毛细管电泳(CE)分离方法. 利用标准试剂混合样考察了缓冲体系的组成、pH值及添加剂对分离的影响,并探讨了尿液中基体成分如肌酸酐、尿酸和乙酰乙酸对分离的干扰. 在Na2B4O7-NaOH缓冲体系(pH=9.90)及紫外(UV)检测(波长200 nm)条件下对8种神经递质类相关物质的分析获得了良好的定量线性关系,检出限(LOD)为0.04~0.60 μmol/L,迁移时间和峰面积的相对标准偏差(RSD,n=5)范围分别为0.09% ~0.48%和0.47% ~3.34%. 利用该方法对实际尿液中的精氨酸和香草扁桃酸进行了定性和定量分析,其定量结果分别为(95.8±3.8)和(44.6±3.5) μmol/L,加标回收率为96.65%~104.5%.

关键词: 毛细管电泳, 神经递质, 尿液, 代谢产物

Abstract:

Capillary electrophoresis(CE) method was developed to separate the mixture of monoamine neurotransmitters(serotonin, dopamine, epinephrine), neurotransmitter metabolites(homovanillic acid, 5-hydroxyl indole acetic acid, vanilmandelic acid) and neurotransmitter precursors(arginine, tyrosine). The compositions, pH and concentration of background electrolyte(BGE) were optimized to separate the standard samples, meanwhile, the possible interferences by matrix components in urine as creatinine, uric acid and acetoacetic acid were investigated. Good quantitative linearity was obtained at the BGE of Na2B4O7-NaOH(pH=9.90) and ultraviolet(UV) detection(200 nm) for eight neurotransmitter-related substances, which owned the limit of detection(LOD) in a range of 0.04—7.00 μmol/L. Relative standard deviation(RSD)(n=5) of migration time and peak area were 0.09%—0.48% and 0.47%—3.34%, respectively. Eventually, the developed CE method was applied for the qualitative and quantitative analysis of arginine(Arg) and vanilmandelic acid(VMA) in urine sample. Their concentrations were respectively at (95.8±3.8) μmol/L and (44.6±3.5) μmol/L with the recovery at a range of 96.65%—104.5%.

Key words: Capillary electrophoresis, Neurotransmitter, Urine, Metabolite

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