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  

Simultaneous determination of amino acid and monoamine neurotransmitters in serum by high performance liquid chromatography coupled with precolumn derivatization

作　　者：白洁 王妲[1] 刘泽平 张佳琪 刘丽艳 韩艳梅[2] BAI Jie;WANG Da;LIU Zeping;ZHANG Jiaqi;LIU Liyan;HAN Yanmei(College of Public Health, Hebei University, Baoding 071000, China;Medical Comprehensive Experimental Center, Hebei University, Baoding 071000, China)

机构地区：[1]河北大学公共卫生学院,河北保定071000 [2]河北大学医学综合实验中心,河北保定071000

出　　处：《色谱》2020年第8期923-928,共6页Chinese Journal of Chromatography

基　　金：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。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),andγ-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and ethanol were mixed at a volume ratio of 1∶2(v/v)for protein precipitation.After centrifugation,the supernatant was withdrawn and blown to dryness using nitrogen.The residue was pre-column derivatized with OPA,and the derivatized product was isolated by gradient elution ona Luna 5u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,5μm).Under the optimal experimental conditions,the five neurotransmitters showed good linearities(r2≥0.9866).The limits of detection were between 0.10 and 0.40μmol/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.

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 

分 类 号：O658[理学—分析化学]