Development of high throughput targeted phospholipidomics based on liquid chromatography-mass spectrometry
10.13699/j.cnki.1001-6821.2018.03.041
- VernacularTitle:基于液质联用技术的高通量靶标磷脂组学的分析研究
- Author:
Bing YANG
1
;
Qiao-Nan LIN
;
Xin XIONG
;
Xian-Hua ZHANG
;
Rong-Sheng ZHAO
Author Information
1. 北京大学第三医院药剂科
- Keywords:
liquid chromatography-tandem mass spectrometry technology;
phospholipidomics;
method validation
- From:
The Chinese Journal of Clinical Pharmacology
2018;34(3):327-331
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a high throughput targeted method to study phospholipids profding and to screen out and quantitate the potential biomarkers in urine samples.Methods The phospholipids in urine was extracted by modified MTBE(methyl tert-butyl ether)method.Semi -quantitative analysis of phospholipids in urine was realized by using high performance liquid chromatography-electrospray ionization-qua-drupoles/trap (HPLC-ESI-Q/Trap) technique.7 standards and 15 endogenous phospholipids were chosen to conduct method validation,including specificity,sensitivity,precision,matrix effect,carryover effect,stability and recovery.Principal component analysis (PCA) of quality control (QC) samples interspersed during the detection process was used to evaluate the reliability of the data obtained.Results The lower limit of quantitation of 7 phospholipids were phosphatidylcholine and lysophosphatidylcholine 0.25 ng· mL-1,phosphatidylethanolamine and phosphatidylserine 2.00 ng· mL-1,phosphatidylglycerol and phosphatidylinositol 1.00 ng · mL-1,sphingomyelin 625.00 pg · mL-1,respectively.During the continuous analysis,the relative standard deviation (RSD) of the retention time was 0.72％-3.44％,the peak area was 0.71％-10.53％.The recoveries of the 7 phospholipids were in the range of 54.05％-105.73％.All samples were stable after being stored 12 h at room temperature,being stored 24h after preparation,two freeze-thaw cycles and being cryopreserved 1 month at-80 ℃.QC samples in the first principal component diagram showed that the data was reliable.Conclusion The developed HPLC-ESI-Q/Trap method was simple,stable and sensitive,which can be applied to the subsequent study of large sample size of phospholipidomics research and quantitative analysis of potential urinary phospholipids biomarkers.
