王雁飞1 陶蓓蓓2 张绪得2 黄春雷3.LC-MS/MS 检测癫痫患者神经递质类氨基酸[J].,2011,11(14):2705-2708 |
LC-MS/MS 检测癫痫患者神经递质类氨基酸 |
Detection of Amino Acids Neurotransmitter in Epilepsy by LC-MS/MS |
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DOI: |
中文关键词: 高效液相色谱- 串联质谱 神经递质 氨基酸 |
英文关键词: LC-MS/MS Neurotransmitter Amino acid |
基金项目:国家自然科学基金资助项目(20905051) |
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中文摘要: |
目的:建立液相色谱串联质谱同位素内标法检测神经递质类氨基酸并用于癫痫患者临床评价。方法:选用AAA-C18 柱色谱
柱,以乙腈水(含有0.01%七氟丁酸、0.1%甲酸)为流动相,采用梯度洗脱进行分离,血浆样品用iTRAQ-115 衍生化试剂处理后,加
入iTRAQ-114 衍生化的氨基酸内标并进样,选用3200QTRAP 型质谱仪的多重反应监测(MRM)扫描方式进行检测。疾病组与健
康组的统计采用t 检验和主成份分析。结果:疾病组和健康组氨基酸测定结果显示:Trp、GABA 两组间没有显著性差异(P>0.05),
Arg、Gly、Ser、Tau、Asp、Glu、EtN、两组间有显著性差异(P<0.05),通过PCA 分析显示,疾病组与健康组之间差异明显,Asp、Glu、
Ser 等是引起差异的主要氨基酸。结论:试验方法灵敏、专属性强,并初步的用于癫痫患者体内氨基酸评价。 |
英文摘要: |
Obiective: To develop a LC-MS/MS method with amino acids isotope as internal standard, to detect amino acids neurotransmitter
in the Epileptic for clinical evaluation. Methods: After derivatizated by iTRAQ-115, plasma samples were added internal
standard which had been derivatizated by iTRAQ-114. The separation was carried out on an AAA-C18 column using acetonitrile and water
(both containing 0.01% heptafluorobutyric acid and 0.1% formic acid) as mobile phase by gradient elution. Detection was carried out
by multiple reaction monitoring (MRM) on a 3200QTRAP LC-MS/MS system. T-test and PCA analysis used as the statistics methods.
Results: There was no significant difference for Trp and GABA and significant difference for Arg, Gly, Ser, Tau, Asp, Glu, EtN between
disease group and health group. There was significant difference between disease group and health group due to Asp, Glu, Ser by PCA
analysis. Conclusion: The method is sensitive and selective and applied to detect amino acids neurotransmitter in the Epileptic for clinical
evaluation. |
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