液相色谱-串联质谱法检测干血点中的同型半胱氨酸

发布时间：2018-05-14 05:24

  本文选题：液相色谱-串联质谱 + 自动移液平台　； 参考：《色谱》2016年06期

【摘要】：建立了一种高通量液相色谱-串联质谱技术检测干血点(DBS)中同型半胱氨酸(homocycteine,Hcy)的方法。以DBS为样本,homocystine-D8为同位素内标,二硫苏糖醇(DTT)为蛋白结合态Hcy的还原剂,使用含0.1%(v/v)甲酸、0.05%(v/v)三氟乙酸的乙腈溶液萃取。整个前处理过程使用自动移液平台及96孔板实现高通量自动化操作。处理后的样本经过Phenomenex CN柱分离,使用多反应监测模式进行LC-MS/MS分析。结果表明:Hcy的检出限为0.12μmol/L(S/N=3),定量限为0.46μmol/L(S/N=10)。Hcy在1.16~148.00μmol/L范围内线性关系良好,R~2=0.994。Hcy的平均回收率为(103.0±4.97)%~(112.0±2.13)%,日内相对标准偏差(RSD)为1.9%~4.6%,日间RSD为1.5%~7.1%。DBS样本在不同温度(-4、-20、22和37℃)下储存不同时间(0、1、2、3、4、5、6、14天)后的稳定性试验显示样本总体RSD15%,经前处理后的样本在48 h内的稳定性试验显示样本总体RSD5%。该方法与传统生化分析方法的相关性好(R~2=0.9818,n=47)。
[Abstract]:A high throughput liquid chromatography-tandem mass spectrometry (HPLC-MS) method was developed for the determination of homocysteine homocyte-homocycteine (Hcyine) in DBS. Homocystine-D8 was used as the internal standard of DBS, DTT was used as the reductant of protein-bound Hcy, and acetonitrile solution containing 0.1% v / v) formic acid was used to extract the acetonitrile solution containing 0.05% V / v) trifluoroacetic acid. The whole preprocessing process uses automatic fluid transfer platform and 96 hole plate to realize high-throughput automatic operation. The treated samples were separated by Phenomenex CN column and analyzed by LC-MS/MS using multi-reaction monitoring model. The results showed that the detection limit was 0.12 渭 mol 路L ~ (-1) 路L ~ (-1) S / N ~ (3) and the quantitative limit was 0.46 渭 mol/L(S/N=10).Hcy. The linear range of 0.46 渭 mol/L(S/N=10).Hcy was 1.16 ~ 148.00 渭 mol/L. The average recovery of 0.994.Hcy was 103.0 卤4.97 ~ 0.994.Hcy was 112.0 卤2.133.The relative standard deviation (RSD) was 1.9 卤4.6. the daily RSD for 1.5%~7.1%.DBS samples was stored at different temperatures (-4-2022 and 37 鈩，

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