α-Synuclein转基因小鼠内源性儿茶酚异喹啉的高效液相色谱-串联质谱检测

发布时间：2018-02-07 11:14

  本文关键词： 帕金森病 Salsolinol N-methyl-salsolinol 多反应监测 α-Synuclein转基因小鼠　出处：《分析科学学报》2017年06期 　论文类型：期刊论文

【摘要】：建立内源性儿茶酚异喹啉类物质1-甲基-6,7-二羟基-1,2,3,4-四氢异喹啉(Salsolinol,Sal)、6,7-二羟基-1,2-二甲基-1,2,3,4-四氢异喹啉(N-methyl-Sal,NMSal)的高效液相色谱-串联质谱(HPLC-MS/MS)定量检测方法,研究α-Synuclein过表达对Sal、NMSal脑内水平的影响。实验优化了色谱及质谱条件,建立基于多反应监测(MRM)技术的Sal和NMSal的高灵敏度、高选择性、高重复性分析方法,并应用此方法对过表达人源野生型(WT)、突变型(A53T)α-Synuclein蛋白的转基因小鼠脑中Sal、NMSal含量进行测定,发现WT、A53T转基因小鼠脑内Sal、NMSal的含量与对照组相比均显著升高。结果表明:Sal、NMSal分别在6.10pmol/L~6.25nmol/L和97.70pmol/L~100.00nmol/L范围呈线性(相关系数r≥0.9995),日内、日间相对标准偏差(RSD)均小于15.0%,回收率分别为86.5%~90.7%和81.0%~91.4%,Sal、NMSal的检出限分别为1.53pmol/L和24.4pmol/L,定量限分别为6.10pmol/L和97.7pmol/L。α-Synuclein过表达会导致内源性神经毒素Sal、NMSal水平的升高,可能与帕金森病发病机制有关。
[Abstract]:A high performance liquid chromatography-tandem mass spectrometry (HPLC-MS) method for the quantitative determination of the endogenous catechol isoquinoline (1-methyl-6-dihydroxy-7-dihydroxyquinoline) Salsolinoline Salsolinoline Salsolinoline-6-dihydroxy-2-dimethyl-1-dimethyl-1-dimethyl-4-tetrahydroisoquinoline (N-methyl-SalNMSaline) was developed, and a high performance liquid chromatography-tandem mass spectrometry (HPLC-MS-MSMS) method was developed for the determination of 1-methyl-6-dihydroquinoline (1-methyl-6-dihydroquinoline) and 4-dihydro-isoquinoline (1-methyl-6-dihydroquinoline). To study the effect of 伪 -Synuclein overexpression on the brain level of Salvia NMSal, the conditions of chromatography and mass spectrometry were optimized, and a high sensitivity, high selectivity and high reproducibility analysis method of Sal and NMSal based on multi-reaction monitoring technique was established. The content of Saline NMSal in the brain of transgenic mice with overexpression of human wild type WTA, mutant A53T) 伪 -Synuclein protein was determined by this method. It was found that the content of Saline NMSal in brain of WTA53T transgenic mice was significantly higher than that of control group. The results showed that the linear range of brain Saline NMSal was 6.10pmol / L 6.25nmol / L and 97.70pmol / L 100.00nmol / L (r 鈮，

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