超高效液相色谱-三重四极杆质谱法测定大鼠海马和大脑皮层中生物胺类及氨基酸类神经化学物质
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摘要
建立了超高效液相色谱-三重四极杆质谱法检测大鼠海马和大脑皮层中生物胺类及氨基酸类神经化学物质含量的方法。选用Hypercarb(100 mm×2.1 mm,5μm)色谱柱,以0.1%甲酸-甲醇为流动相,梯度洗脱。电喷雾离子源(ESI)在正离子模式下,选择多反应监测模式(MRM)扫描,对检测的6种生物胺类和11种氨基酸类神经化学物质分别选择一个定性离子对和一个定量离子对。测定Wistar大鼠海马和大脑皮层中6种生物胺类和11种氨基酸类神经化学物质的含量,对比两种不同脑组织样品前处理方法对神经化学物质含量测定结果的影响。17种神经化学物质在10 min内即可完成同时测定,检测方法线性关系良好,日内和日间精密度、加标回收率和重复性满足分析要求。本方法准确、灵敏度高、专属性强、重复性好,适用于脑组织中生物胺类及氨基酸类神经化学物质的含量测定,可为生物胺类及氨基酸类神经化学物质提供有效的样品前处理方法和检测手段。
An ultra performance liquid chromatography-tandem mass(UPLC-MS / MS)spectrometry method was established for determination of the contents of bioamine and amino acid neurochemicals in hippocampus and cerebral cortex of rat. Hypercarb column(100 mm × 2. 1 mm,5 μm)was used for the sample separation with 0. 1% formic acid-methanol as the mobile phase under gradient elution. The neurochemicals were detected by MS / MS using ESI ion source under positive ionization mode with MRM scan. Both quantification and confirmation ions were chosen for each 6 bioamine and 11 amino acid neurochemicals. The influence of two different processing methods on the contents of neurochemicals in brain tissues was compared. Total 17 neurochemicals were simultaneously detected in 10 min. The calibration curve was with a good linear relationship. The intra-day and inter-day precision,average recovery and repeatability could meet the analysis requirement. This UPLC-MS / MS method shows excellent selectivity,accuracy,high sensitivity,specificity and good repeatability,and is suitable for the separation and quantization of bioamine and amino acid neurochemicals in brain tissue.
An ultra performance liquid chromatography-tandem mass(UPLC-MS / MS)spectrometry method was established for determination of the contents of bioamine and amino acid neurochemicals in hippocampus and cerebral cortex of rat. Hypercarb column(100 mm × 2. 1 mm,5 μm)was used for the sample separation with 0. 1% formic acid-methanol as the mobile phase under gradient elution. The neurochemicals were detected by MS / MS using ESI ion source under positive ionization mode with MRM scan. Both quantification and confirmation ions were chosen for each 6 bioamine and 11 amino acid neurochemicals. The influence of two different processing methods on the contents of neurochemicals in brain tissues was compared. Total 17 neurochemicals were simultaneously detected in 10 min. The calibration curve was with a good linear relationship. The intra-day and inter-day precision,average recovery and repeatability could meet the analysis requirement. This UPLC-MS / MS method shows excellent selectivity,accuracy,high sensitivity,specificity and good repeatability,and is suitable for the separation and quantization of bioamine and amino acid neurochemicals in brain tissue.
引文
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23 LI Xue,PI Zi-Feng,XING Jun-Peng,LIN Na,LIU Zhi-Qiang,SONG Feng-Rui.Chinese J.Anal.Chem.,2014,42(11):1646-1650李雪,皮子凤,邢俊鹏,林娜,刘志强,宋凤瑞.分析化学,2014,42(11):1646-1650
24 WANG Qian-Qian,ZHANG Jing,PI Zi-Feng,LIU Shu,SONG Feng-Rui,LIU Zhi-Qiang.Chinese J.Anal.Chem.,2014,42(7):997-1001王倩倩,张静,皮子凤,刘舒,宋凤瑞,刘志强.分析化学,2014,42(7):997-1001
25 ANZuo-Ling,SHI Chen,ZHAO Rui,LI Peng-Fei,LIU Li-Hong.Chinese J.Anal.Chem.,2015,43(9):1408-1414安卓玲,史忱,赵瑞,李鹏飞,刘丽宏.分析化学,2015,43(9):1408-1414
26 Falasca S,Petruzziello F,Kretz R,Rainer G,Zhang X Z.J.Chromatogr.A,2012,1241:46-51
2 Brichta L,Greengard P,Flajolet M.Trends Neurosci.,2013,36:543-554
3 Carlsson A,Waters N,Carlsson M L.Eur.Arch.Psychiatry Clin.Neurosci.,1999,249(Suppl.4):37-43
4 Kenche V B,Zawisza I,Masters C L,Bal W,Barnham K J,Drew S C.Inorg.Chem.,2013,52:4303-4318
5 Zhao L,Zheng S,Su G,Lu X,Yang J,Xiong Z,Wu C.J.Chromatogr.B,2015,988:59-65
6 LIU Tian-Ya,HONG Zong-Yuan,QU Wei-Min,HUANG Zhi-Li.Acta Pharmaceutica Sinica,2011,46(3):247-252刘天雅,洪宗元,曲卫敏,黄志力.药学学报,2011,46(3):247-252
7 PI Zi-Feng,WANG Qian-Qian,ZHANG Jing,Song Feng-Rui,LIU Zhi-Qiang.Chem.J.Chinese Universities,2015,36(3):422-448皮子凤,王倩倩,张静,宋凤瑞,刘志强.高等学校化学学报,2015,36(3):422-448
8 JING Hong-Jiang,CHENG Yi-Yong,LI Shu-Tian,ZHANG Guang-Yuan.Journal of Hygiene Research,2000,29(1):40-42景洪江,程义勇,李树田,章广远.卫生研究,2000,29(1):40-42
9 ZHANG Wei,LIU Peng,YUAN Yuan,ZHANG Yi-Bao,ZHANG Shao-Bo,MA Jing-Lin,WANG Jing.Chinese Journal of Cerebrovascular Diseases,2000,29(1):40-42张伟,刘鹏,袁媛,张义宝,张少博,马靖琳,汪静.中国脑血管病杂志,2016,13(4):198-203
10 YANG Pei,LIXue-Chun,TIAN Jing-Chen,LIN Long-Fei,FU Jing,ZHANG Hui,NI Jian.World Science and Technology/Modernization of Traditional Chinese Medicine and Materia Medica,2014,16(10):2174-2179杨培,李雪春,田晶辰,林龙飞,付京,张慧,倪健.世界科学技术-中医药现代化,2014,16(10):2174-2179
11 YANG Xiao-Ting,HU Yu-Fei,LI Gong-Ke.Journal of Instrumental Analysis,2013,32(11):1394-1400杨晓婷,胡玉斐,李攻科.分析测试学报,2013,32(11):1394-1400
12 GU Qun,SHI Xian-Zhe,XU Guo-Wang.Chinese Journal of Chromatography,2007,25(4):457-462顾群,石先哲,许国旺.色谱,2007,25(4):457-462
13 Bucher E S,Wightman R M.Annu.Rev.Anal.Chem.,2015,8:239-261
14 LI Hui,LI Zhe-Xuan,ZHANG Yuan,LUO Wen-Hong.Journal of Instrumental Analysis,2005,24(2):89-91李慧,林哲绚,张原,罗文鸿.分析测试学报,2005,24(2):89-91
15 SUFeng-Li,WANG Feng,ZHU Rong-Hua,HUANG Ai-Ling,LI Huan-De.Chin.J.Pharm.Anal.,2008,28(8):1238-1243苏芬丽,王峰,朱荣华,黄爱玲,李焕德.药物分析杂志,2008,28(8):1238-1243
16 Wei B,Li Q,Fan R,Su D,Chen X,Jia Y,Bi K.J.Pharm.Biomed.Anal.,2014,88:416-422
17 Cai H L,Zhu R H,Li H D,Zhang J,Li L F.J.Chromatogr.B,2011,879:1993-1999
18 Zhao X E,Suo Y R.Talanta,2008,76:690-697
19 Cai H L,Zhu R H,Li H D.Anal.Biochem.,2010,396:103-111
20 ZHAO Xian-En,YOU Jin-Mao,LIU Hong-Zhen,SUO You-Rui.Chinese J.Anal.Chem.,2007,35(7):938-944赵先恩,尤进茂,刘洪珍,索有瑞.分析化学,2007,35(7):938-944
21 ZHAO Xian-En,SUO You-Rui.Chinese J.Anal.Chem.,2008,36(1):12-18赵先恩,索有瑞.分析化学,2008,36(1):12-18
22 Yang Z L,Li H,Wang B,Liu S Y.J.Chromatogr.B,2016,1012-1013:79-88
23 LI Xue,PI Zi-Feng,XING Jun-Peng,LIN Na,LIU Zhi-Qiang,SONG Feng-Rui.Chinese J.Anal.Chem.,2014,42(11):1646-1650李雪,皮子凤,邢俊鹏,林娜,刘志强,宋凤瑞.分析化学,2014,42(11):1646-1650
24 WANG Qian-Qian,ZHANG Jing,PI Zi-Feng,LIU Shu,SONG Feng-Rui,LIU Zhi-Qiang.Chinese J.Anal.Chem.,2014,42(7):997-1001王倩倩,张静,皮子凤,刘舒,宋凤瑞,刘志强.分析化学,2014,42(7):997-1001
25 ANZuo-Ling,SHI Chen,ZHAO Rui,LI Peng-Fei,LIU Li-Hong.Chinese J.Anal.Chem.,2015,43(9):1408-1414安卓玲,史忱,赵瑞,李鹏飞,刘丽宏.分析化学,2015,43(9):1408-1414
26 Falasca S,Petruzziello F,Kretz R,Rainer G,Zhang X Z.J.Chromatogr.A,2012,1241:46-51