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柱前衍生-高效液相色谱法同时测定血清中氨基酸类及单胺类神经递质
引用本文:白洁,王妲,刘泽平,张佳琪,刘丽艳,韩艳梅.柱前衍生-高效液相色谱法同时测定血清中氨基酸类及单胺类神经递质[J].色谱,2020,38(8):923-928.
作者姓名:白洁  王妲  刘泽平  张佳琪  刘丽艳  韩艳梅
作者单位:1 河北大学公共卫生学院, 河北 保定 0710002 河北大学医学综合实验中心, 河北 保定 071000
基金项目:国家自然科学基金(21605035);河北大学校内人才培养科研项目(521000981384)
摘    要:以邻苯二甲醛(o-phthalaldehyde,OPA)为衍生试剂,建立了柱前衍生-高效液相色谱(HPLC)同时测定血清中氨基酸类神经递质牛磺酸(Tau)、谷氨酸(Glu)、甘氨酸(Gly)、γ-氨基丁酸(γ-GABA)和单胺类神经递质多巴胺(DA)含量的分析方法。血清与乙醇以1∶2的体积比混合,进行蛋白质沉淀后离心,取其上清液,氮吹至近干。前处理后的样品与OPA进行柱前衍生,衍生化产物采用Luna 5u C18色谱柱(250 mm×4.6 mm,5μm)分离,以柠檬酸-乙酸钠缓冲溶液(pH 3.73)为流动相A、乙腈为流动相B进行梯度洗脱,流速为1.0 mL/min,柱温为30℃,检测波长为338 nm。5种神经递质在各自范围内线性关系良好(r2≥0.9866),检出限为0.10~0.40μmol/L,不同加标水平下目标物的加标回收率为87.57%~115.31%,相对标准偏差均低于7.80%。方法操作简单,灵敏度高,精密度、线性关系和回收率等方法学指标较好,可实现血清中氨基酸类及单胺类神经递质的同时检测。

关 键 词:高效液相色谱  柱前衍生  氨基酸类神经递质  单胺类神经递质  邻苯二甲醛  血清
收稿时间:2019-12-23

Simultaneous determination of amino acid and monoamine neurotransmitters in serum by high performance liquid chromatography coupled with precolumn derivatization
BAI Jie,WANG Da,LIU Zeping,ZHANG Jiaqi,LIU Liyan,HAN Yanmei.Simultaneous determination of amino acid and monoamine neurotransmitters in serum by high performance liquid chromatography coupled with precolumn derivatization[J].Chinese Journal of Chromatography,2020,38(8):923-928.
Authors:BAI Jie  WANG Da  LIU Zeping  ZHANG Jiaqi  LIU Liyan  HAN Yanmei
Institution:1 College of Public Health, Hebei University, Baoding 071000, China2 Medical Comprehensive Experimental Center, Hebei University, Baoding 071000, China
Abstract:Using o-phthalaldehyde (OPA) as the derivatization reagent, a precolumn derivatized -high performance liquid chromatography (HPLC) method was developed for the simultaneous determination of amino acid neurotransmitters taurine (Tau), glutamic acid (Glu), glycine(Gly), and γ -aminobutyric acid (γ-GABA), as well as the monoamine neurotransmitter dopamine (DA), in serum samples. The samples and ethanol were mixed at a volume ratio of 1:2 (v/v) for protein precipitation. After centrifugation, the supernatant was withdrawn and blown to dryness using nitrogen. The residue was pre-column derivatized with OPA, and the derivatized product was isolated by gradient elution ona Luna 5u C18 column (250 mm×4.6 mm, 5 μm). Under the optimal experimental conditions, the five neurotransmitters showed good linearities (r2 ≥ 0.9866). The limits of detection were between 0.10 and 0.40 μmol/L. The spiked recoveries at different spiked levels were 87.57%-115.31%, and the RSDs were below 7.80%. This method is simple, sensitive, and it can be promised for the simultaneous detection of amino acid and monoamine neurotransmitters.
Keywords:high performance liquid chromatography (HPLC)  precolumn derivatization  amino acid neurotransmitter  monoamine neurotransmitters  o-phthalaldehyde (OPA)  serum  
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