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. doi: 10.3724/SP.J.1123.2019.12029 shu

Citation: 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. doi: 10.3724/SP.J.1123.2019.12029 shu

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

白洁 ^{1,2,
,} ,
王妲 ^1, ,
刘泽平 ^1, ,
张佳琪 ^1, ,
刘丽艳 ^2, ,
韩艳梅 ^{2,
,}

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

通讯作者: 白洁, E-mail:bj0427@126.com; 韩艳梅, E-mail:hanyanmei6465@sina.com

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，5 μm）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0.10~0.40 μmol/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:

English

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

BAI Jie ^{1,2,
,} ,
WANG Da ^1, ,
LIU Zeping ^1, ,
ZHANG Jiaqi ^1, ,
LIU Liyan ^2, ,
HAN Yanmei ^{2,
,}

1.
College of Public Health, Hebei University, Baoding 071000, China;
2.
Medical Comprehensive Experimental Center, Hebei University, Baoding 071000, China

Corresponding author: BAI Jie, ; HAN Yanmei,

Received Date: 23 December 2019
Fund Project:
National Natural Science Foundation of China (No. 21605035)

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 After centrifugation, the supernatant was withdrawn and blown to dryness using nitrogen.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.

Key words:

high performance liquid chromatography (HPLC)
/ precolumn derivatization
/ amino acid neurotransmitter
/ monoamine neurotransmitters
/ o-phthalaldehyde (OPA)
/ serum

1. [1] Si B, Song E. Chemosensors, 2018, 6(1): 1
2. [2] Brandman O, Liou J, Park W S, et al. Cell, 2007, 131(7): 1327
3. [3] Sangubotla R, Kim J. TrAC-Trends Anal Chem, 2018, 105: 240
4. [4] Meller S, Brandt C, Theilmann W, et al. Brain Res, 2019, 1712: 109
5. [5] Azmanova M, Pitto-Barry A, Barry N P E. Med Chem Comm, 2018, 9(5): 759
6. [6] Mayor D, Tymianski M. Neuropharmacology, 2018, 134: 178
7. [7] Stead S M, Daube J R. Clinical Neurophysiology, Oxford: Oxford University Press, 2016
8. [8] Luan H, Huang Y, Li J, et al. Microgravity Sci Technol, 2018, 30(4): 483
9. [9] Forgacsova A, Galba J, Garruto R M, et al. JChromatogr B, 2018, 1073: 154
10. [10] Kuster A, Arnoux J B, Barth M, et al. J Inherit Metab Dis, 2018, 41(1): 129
11. [11] Cao Y H, Zhang M Y, Liu M, et al. Chinese Journal of Chromatography, 2019, 37(3): 265 曹雨虹, 张明勇, 刘敏, 等. 色谱, 2019, 37(3): 265
12. [12] Zhao Y Y, Liu L Y, Han Y Y, et al. Chinese Journal of Chromatography, 2011, 29(2): 146 赵燕燕, 刘丽艳, 韩媛媛, 等. 色谱, 2011, 29(2): 146
13. [13] Qi W M, Zhao X E, Qi Y, et al. Chinese Journal of Chromatography, 2015, 33(9): 981 亓伟梅, 赵先恩, 亓永, 等. 色谱, 2015, 33(9): 981
14. [14] Sun Z, Li Q, Bi K. Molecules, 2018, 23(12): 3111
15. [15] Huang D, Jiang Q S, Yang J Q, et al. Biomed Chromatogr, 2018, 32(11): e4338
16. [16] Olesti E, Rodríguez-Morató J, Gomez-Gomez A, et al. Talanta, 2019, 192: 93
17. [17] Kuang H X, Zhang H B, Tan J H, et al. Chinese Journal of Chromatography, 2019, 37(4): 404 邝洪轩, 张海彬, 谭建华, 等. 色谱, 2019, 37(4): 404
18. [18] Hook V, Kind T, Podvin S, et al. ACS Chem Neurosci, 2019, 10(3): 1369
19. [19] Sánchez-López E, Montealegre C, Crego A L, et al. TrAC-Trends Anal Chem, 2015, 67: 82
20. [20] Madhu S, Manickam P, Pierre M, et al. Sens Actuator A: Phys, 2018, 269: 401
21. [21] Vander Ende E, Bourgeois M C, Henry A I, et al. Anal Chem, 2019, 91(15): 9554
22. [22] Arya A, Sindhwani G, Kadian R. Asian J Pharm Clin Res, 2018, 11(6): 4
23. [23] Li H, Lu Y J, Zheng X X. Journal of Chinese Microcirculation, 2003, 7(3): 160 李惠, 卢英俊, 郑筱祥. 中国微循环, 2003, 7(3): 160
24. [24] Chen L M, Shang Y F, Zhao M B, et al. Chinese Journal of Chromatography, 2010, 128(12): 1154 陈丽梅, 尚艳芬, 赵孟彬, 等. 色谱, 2010, 128(12): 1154
25. [25] He X R, Wu Z Y, Ye Y L, et al. Journal of Instrumental Analysis, 2016, 35(7): 922 赫欣睿, 武中庸, 叶永丽, 等. 分析测试学报, 2016, 35(7): 922
26. [26] Zhao F, Li Q, Liang M L, et al. Chinese Journal of Analytical Chemistry, 2018, 46(1): 121 赵芳, 李强, 梁梅丽, 等. 分析化学, 2018, 46(1): 121
27. [27] Li H, Lu W H, Jiao L L, et al. Chinese Journal of Analytical Chemistry, 2018, 46(4): 586 李慧, 鲁文慧, 焦丽丽, 等. 分析化学, 2018, 46(4): 586

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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

通讯作者: 白洁, E-mail:bj0427@126.com; 韩艳梅, E-mail:hanyanmei6465@sina.com

English

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

Corresponding author: BAI Jie, ; HAN Yanmei,

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中国化学会秘书处

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

通讯作者: 白洁, E-mail:bj0427@126.com; 韩艳梅, E-mail:hanyanmei6465@sina.com

English

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

Corresponding author: BAI Jie, ; HAN Yanmei,

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

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