维生素C抗氧化-同位素内标稀释-超高效液相色谱-串联质谱同时测定新生期大鼠海马体神经递质的含量

引用本文: 邝洪轩, 张海彬, 谭建华, 雒亦凡, 刘舒华, 庞启华, 李楚华, 范瑞芳. 维生素C抗氧化-同位素内标稀释-超高效液相色谱-串联质谱同时测定新生期大鼠海马体神经递质的含量[J]. 色谱, 2019, 37(4): 404-411. doi: 10.3724/SP.J.1123.2018.11012 shu

Citation: KUANG Hongxuan, ZHANG Haibin, TAN Jianhua, LUO Yifan, LIU Shuhua, PANG Qihua, LI Chuhua, FAN Ruifang. Simultaneous determination of five neurotransmitters in neonatal rat hippocampus by adding vitamin C coupled with isotope dilution-ultra-high performance liquid chromatography-tandem mass spectrometry[J]. Chinese Journal of Chromatography, 2019, 37(4): 404-411. doi: 10.3724/SP.J.1123.2018.11012 shu

维生素C抗氧化-同位素内标稀释-超高效液相色谱-串联质谱同时测定新生期大鼠海马体神经递质的含量

1.
华南师范大学生命科学学院, 广东省药食资源生物加工及综合利用工程技术研究中心, 广东广州 510631;
2.
广东工业大学环境科学与工程系, 广州市环境催化与污染控制重点实验室, 广东广州 510006;
3.
广州质量监督检测研究院, 广东广州 511447
基金项目:
国家自然科学基金项目（21777048）；广州市环境催化与污染控制重点实验室开放基金（GKLECPC-09）.

摘要: 脑区神经递质的测定对于研究神经系统的作用机制具有重要意义。该研究建立了同位素内标稀释-超高效液相色谱-串联质谱同时测定双酚A暴露后大鼠海马体中5种神经递质含量的方法，包括谷氨酸、γ-氨基丁酸、乙酰胆碱、多巴胺以及5-羟色胺。选用2%（体积分数）乙酸水-甲醇（9：1，v/v）溶液配制标准样品及溶解样品，以Ultimate AQ-C₁₈（150 mm×4.6 mm，3 μ m）为分离柱，0.1%（体积分数）甲酸水和甲醇为流动相，在28℃柱温下，梯度洗脱，所有化合物在3 min内出峰。添加维生素C能显著减少多巴胺和5-羟色胺在前处理以及样品储存过程中的氧化，极大地提高神经递质的稳定性，从而准确定量。该方法标准曲线线性关系良好，相关系数R²>0.998，检出限和定量限低，日内和日间精密度为0.39%~13.6%，加标回收率为92.9%~119%，实际进样残留低，已被成功地应用于新生期双酚A暴露后大鼠海马体神经递质含量的测定。

关键词:

English

Simultaneous determination of five neurotransmitters in neonatal rat hippocampus by adding vitamin C coupled with isotope dilution-ultra-high performance liquid chromatography-tandem mass spectrometry

1.
Guangdong Provincial Engineering Technology Research Center for Drug and Food Biological Resources Processing and Comprehensive Utilization, School of Life Sciences, South China Normal University, Guangzhou 510631, China;
2.
Guangzhou Key Laboratory of Environmental Catalysis and Pollution Control, School of Environmental Science and Engineering, Institute of Environmental Health and Pollution Control, Guangdong University of Technology, Guangzhou 510006, China;
3.
GuangZhou Quality Supervision and Testing Institute, Guangzhou 511447, China
Received Date: 08 November 2018
Fund Project:
National Natural Science Foundation of China (No. 21777048)

Abstract: Analysis of neurotransmitters in brain tissue is useful for mechanistic studies of the central nervous system. Isotope dilution coupled with ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was developed for the simultaneous determination of glutamic acid, γ-amino butyric acid, acetylcholine, dopamine, and serotonin in the hippocampus tissue. A 2% (v/v) acetic acid in water-methanol (9:1, v/v) solution was used to prepare the standards and re-dissolute samples after nitrogen drying. An Ultimate AQ-C₁₈ column (150 mm×4.6 mm, 3 μm) was used as the analytical column and 0.1% (v/v) formic acid in water and methanol were used as the mobile phase. Gradient elution was performed and all target compounds were eluted over 3 min at 28℃. Vitamin C spiked during the sample pretreatment and storage periods significantly retarded the oxidation of dopamine and serotonin and improved the neurotransmitter stability. The developed method showed good linearities (correlation coefficient (R²)>0.998), low detection limits (0.15-1.0 μg/L), good inter-and intra-day precisions (relative standard deviations:0.39%-13.6%), good accuracy (92.9%-119%), low carry-over, and excellent stability. Moreover, the method was successfully applied and validated in the determination of rat hippocampus exposed to bisphenol A.

Key words:

ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS)
/ neurotransmitters
/ vitamin C
/ hippocampus

1. [1] Huang X, Li S P, Zhang Y, et al. Chinese Journal of Analytical Chemistry, 2016, 44(11):1652 黄鑫, 李帅坪, 张勇, 等. 分析化学, 2016, 44(11):1652
2. [2] Bergh M S, Bogen I L, Lundanes E. J Chromatogr B Analyt Technol Biomed Life Sci, 2016, 1028:120
3. [3] Wu X, Jiang Q, Wang S, et al. Biomed Chromatogr, 2014, 28(4):459
4. [4] 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
5. [5] Zhang L, Lu Y J, Deng T L, et al. Chinese Journal of Chromatography, 2005, 23(1):76 张雷, 卢英俊, 邓同乐, 等. 色谱, 2005, 23(1):76
6. [6] Yoshitake T, Fujino K, Kehr J, et al. Anal Biochem, 2003, 312(2):125
7. [7] Hou J G, Liu H L, He T X, et al. Chinese Journal of Chromatography, 2002, 20(2):140 侯经国, 刘慧玲, 何天稀, 等. 色谱, 2002, 20(2):140
8. [8] Del Pino J, Martinez M A, Castellano V J, et al. Toxicology, 2011, 287:145
9. [9] Parrot S, Neuzeret P C, Denoroy L, et al. J Chromatogr B Analyt Technol Biomed Life Sci, 2011, 879:3871
10. [10] 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
11. [11] 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
12. [12] Wojnicz A, Ortiz J A, Casas A, et al. Clinica Chimica Acta, 2016, 453:174
13. [13] Greco S, Danysz W, Zivkovic A, et al. Anal Chim Acta, 2013, 771:65
14. [14] Zhang M L, Fang C W, Smagin G. J Pharm Biomed Anal, 2014, 100:357
15. [15] European Medicines Agency (EMA). Guideline on Bioanalytical Method Validation EMEA/CHMP/EWP/192217/2009. (2011-07-21)[2018-10-09]. https://wenku.baidu.com/view/6f5bfa0852ea551810a68749.html
16. [16] Zhou Y X, Wang Z Y, Xia M H, et al. Environ Pollut, 2017, 229:40

扫一扫看文章

计量

PDF下载量: 4
文章访问数: 738
HTML全文浏览量: 82

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

维生素C抗氧化-同位素内标稀释-超高效液相色谱-串联质谱同时测定新生期大鼠海马体神经递质的含量

English

Simultaneous determination of five neurotransmitters in neonatal rat hippocampus by adding vitamin C coupled with isotope dilution-ultra-high performance liquid chromatography-tandem mass spectrometry

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？

计量

通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

维生素C抗氧化-同位素内标稀释-超高效液相色谱-串联质谱同时测定新生期大鼠海马体神经递质的含量

English

Simultaneous determination of five neurotransmitters in neonatal rat hippocampus by adding vitamin C coupled with isotope dilution-ultra-high performance liquid chromatography-tandem mass spectrometry

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？

计量

文章相关

通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs