Establishment and Application of High Throughput Analysis Method for Methylcardiolipin Based on High Resolution Mass Spectrometry

JIAO Yu-pei; WANG Xue-ying; XU Li-na; WANG Yu-song; LIU Zi-qi; LIU Xiao-hui

doi:10.19969/j.fxcsxb.21100601

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Establishment and Application of High Throughput Analysis Method for Methylcardiolipin Based on High Resolution Mass Spectrometry

Scientific Paper | 更新时间：2022-06-22

- Establishment and Application of High Throughput Analysis Method for Methylcardiolipin Based on High Resolution Mass Spectrometry
- Journal of Instrumental Analysis Vol. 41, Issue 5, Pages: 717-723(2022)
- 作者机构：
  
  1.清华大学蛋白质研究技术中心，北京 100084
  2.清华大学生命学院，北京 100084
- 作者简介：
- 基金信息：
- DOI：10.19969/j.fxcsxb.21100601
  CLC：
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焦玉佩,王雪颖,许丽娜等.基于高分辨质谱的甲基化心磷脂高通量分析方法的建立和应用[J].分析测试学报,2022,41(05):717-723.

JIAO Yu-pei,WANG Xue-ying,XU Li-na,et al.Establishment and Application of High Throughput Analysis Method for Methylcardiolipin Based on High Resolution Mass Spectrometry[J].Journal of Instrumental Analysis,2022,41(05):717-723.
焦玉佩,王雪颖,许丽娜等.基于高分辨质谱的甲基化心磷脂高通量分析方法的建立和应用[J].分析测试学报,2022,41(05):717-723. DOI： 10.19969/j.fxcsxb.21100601.

JIAO Yu-pei,WANG Xue-ying,XU Li-na,et al.Establishment and Application of High Throughput Analysis Method for Methylcardiolipin Based on High Resolution Mass Spectrometry[J].Journal of Instrumental Analysis,2022,41(05):717-723. DOI： 10.19969/j.fxcsxb.21100601.

摘要

该文基于心磷脂标准品的甲基化反应，建立了高灵敏度高通量分析甲基化心磷脂的液相色谱 - 高分辨质谱方法。由于甲基化后的心磷脂具有特征性的碎片，因此建立了包含780个分子特征片段的模拟数据库用于脂质鉴定。衍生化后，CL（14∶0/14∶0/14∶0/14∶0）信号提高了10倍，定量下限（LOQ）达到10 ng/mL。将该方法应用于小鼠衰老过程中心磷脂的调控研究，通过Tracefinder本地二级数据库共鉴定了43个心磷脂分子，其中21个心磷脂在衰老的大脑中有差异变化。结果发现：含有FA（20∶4）、FA（22∶6）长链多不饱和脂肪酸的心磷脂在衰老过程中呈下降趋势。高分辨质谱平台为甲基化的心磷脂分析提供了一种高通量筛选方法，对于低丰度心磷脂的检测具有很大潜力。

Abstract

Cardiolipin is important for assembly and integrity of mitochondrial electron transfer complex．A sensitive and high throughput method via liquid chromatography - high resolution mass spectrometry was developed for the detection of low abundant cardiolipin based on methylation reaction．Unique fragmentation pattern was derived from methylated cardiolipins．Therefore，a simulated database including the featured fragments of 780 molecules was established for lipid identification．After derivatization，the signal of CL（14∶0/14∶0/14∶0/14∶0） was increased by 10 times，and 10 ng/mL of the limit of quantitation（LOQ） was achieved in the analysis．This method was applied to study the cardiolipin regulation in mouse brain during aging．A total of 43 cardiolipin molecules were identified using Tracefinder coupled with in-house MS/MS database，of which 21 cardiolipin molecules were differentially changed in aged brains．Cardiolipins with polyunsaturated acyl-component e.g. FA（20∶4） and FA（22∶6） were decreased during the aging process．The orbitrap-based platform with methylation provides a high throughput screening approach for cardiolipin analysis，which is potentially beneficial to the detection of low abundant lipids．

关键词

心磷脂甲基化高分辨质谱高通量

Keywords

cardiolipinmethylationhigh resolution mass spectrometryhigh throughput

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