中国中药杂志

2019, v.44(22) 4918-4923

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UPLC-MS/MS同时快速检测大鼠组织及血、尿样品中8种神经递质含量
Simultaneous rapid detection of eight neurotransmitters in rat tissues,blood and urine samples by UPLC-MS/MS

毕欣宁;戚婧;黄运芳;魏桂杰;张婷;王宏雅;王淑琪;李志慧;郑伟;汪祺;张玉杰;
BI Xin-ning;QI Jing;HUANG Yun-fang;WEI Gui-jie;ZHANG Ting;WANG Hong-ya;WANG Shu-qi;LI Zhi-hui;ZHENG Wei;WANG Qi;ZHANG Yu-jie;School of Chinese Materia Medica,Bejing University of Chinese Medicine;National Institutes for Food and Drug Control;

摘要(Abstract):

建立超高效液相色谱-串联四极杆质谱(UPLC-MS/MS)同时测定大鼠脑、肝、肾、肾上腺、血清和尿液中8种神经递质的含量,包括五羟色胺、五羟吲哚乙酸、肾上腺素、去甲肾上腺素、多巴胺、谷氨酸、γ-氨基丁酸、乙酰胆碱,并以此考察神经递质在大鼠组织和血、尿中的分布特性。选用Waters ACQUITY UPLC BEH C_(18)色谱柱(2. 1 mm×100 mm,1. 7μm),以0. 3%甲酸水-乙腈作为流动相进行梯度洗脱,采用大气压电喷雾离子源(ESI),正离子模式,多反应监测(MRM)扫描方式建立神经递质的液质方法并进行方法学研究;将正常大鼠血浆、尿及组织经匀浆、离心、除蛋白等前处理后的上清液2μL进样分析。结果显示8种神经递质可在7 min内准确测定,线性相关系数均大于0. 99,准确度高,精密度良好,专属性、稳定性、提取回收率和基质效应等均符合生物样品分析要求;在大鼠脑、肝、肾、肾腺、血、尿中含量最多的递质分别是γ-氨基丁酸、谷氨酸、谷氨酸、肾上腺素、谷氨酸和多巴胺。该方法灵敏快捷、精密准确、专属稳定,可用于生物样品中8种神经递质的同时定量分析。考察递质在大鼠体内的分布比例关系对疾病的防治具有重要意义。
Ultra performance liquid chromatography-tandem mass spectrometry( UPLC-MS/MS) was used to establish the simultaneous determination method of eight neurotransmitters in brain,liver,kidney,adrenal gland,serum and urine,including serotonin,5-hydroxyindole acetic acid,epinephrine,norepinephrine,dopamine,glutamic acid,γ-aminobutyric acid,and acetylcholine,and then investigate the distribution characteristics of neurotransmitters in rat tissues,blood and urine. Waters ACQUITY UPLC BEH C_(18) column( 2. 1 mm×100 mm,1. 7 μm) was used,with 0. 3% formic acid solution-acetonitrile as the mobile phase for gradient elution.Multiple reaction monitoring( MRM) scanning method under positive mode by atmospheric pressure electrospray ion source( ESI) was also performed to establish the detection method of neurotransmitters for methodological research. The plasma,urine and tissues of normal rats were pre-treated including homogenization,centrifuging,and protein removal,then the 2 μL supernatant was injected for analysis. The results showed that eight kinds of neurotransmitters could be accurately determined within 7 min,with linear correlation coefficients all greater than 0. 99. This method showed high accuracy and good precision,with specificity,stability,extraction recovery and matrix effects all complying with the biological sample analysis requirements. The most abundant transmitters in the brain,liver,kidney,kidney gland,blood and urine were γ-aminobutyric acid,glutamic acid,glutamic acid,adrenaline,glutamic acid and dopamine.The method is sensitive,rapid,precise,accurate and specific,and can be used for simultaneous quantitative analysis of eight neurotransmitters in biological samples. The investigation of the distribution ratio of transmitters in rats is of important significance to disease prevention and treatment.

关键词(KeyWords): UPLC-MS/MS;神经递质;含量测定;生物样品;分布特性
UPLC-MS/MS;neurotransmitters;content determination;biological samples;distribution characteristics

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(81673680)

作者(Author): 毕欣宁;戚婧;黄运芳;魏桂杰;张婷;王宏雅;王淑琪;李志慧;郑伟;汪祺;张玉杰;
BI Xin-ning;QI Jing;HUANG Yun-fang;WEI Gui-jie;ZHANG Ting;WANG Hong-ya;WANG Shu-qi;LI Zhi-hui;ZHENG Wei;WANG Qi;ZHANG Yu-jie;School of Chinese Materia Medica,Bejing University of Chinese Medicine;National Institutes for Food and Drug Control;

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DOI: 10.19540/j.cnki.cjcmm.20190428.502

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