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| Establishment and Application of High Throughput Analysis Method for Methylcardiolipin Based on High Resolution Mass Spectrometry |
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| DOI： |
| KeyWord:cardiolipin ?methylation ?high resolution mass spectrometry ?high throughput |
| Author | Institution |
| JIAO Yu-pei，WANG Xue-ying，XU Li-na，WANG Yu-song，LIU Zi-qi，LIU Xiao-hui |
1. National Protein Science Facility（Beijing），Tsinghua University，Beijing ，China;2. School of Life Sciences，Tsinghua University，Beijing ，China |
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| 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． |
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