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Quantitative analysis of phosphatidylcholine in rat liver tissue by nanoflow liquid chromatography/tandem mass spectrometry
Author(s) -
Ahn Eun Jeong,
Kim Hanna,
Chung Bong Chul,
Moon Myeong Hee
Publication year - 2007
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.200700231
Subject(s) - degree of unsaturation , chemistry , chromatography , quantitative analysis (chemistry) , calibration curve , phosphatidylcholine , mass spectrometry , liquid chromatography–mass spectrometry , tandem mass spectrometry , carbon chain , analytical chemistry (journal) , phospholipid , membrane , detection limit , biochemistry , organic chemistry
A quantitative method was developed for the determination of phosphatidylcholine (PC) species concentration using nanoflow LC‐ESI‐MS/MS. In this study, a calibration method is developed to determine the effect of PC carbon chain length on MS peak intensity. Using the multiple standard addition method, a relationship between the peak intensities of different PC species from nanoflow LC‐MS and carbon chain length is established first using different injection amounts of PC standards. From this relationship, a calibration curve for each carbon chain length can be obtained for the concentration calculation. It was found that the MS peak area of PC species analyzed by nanoflow LC‐MS linearly decreased with increased acyl carbon numbers, and that the effect of the degree of acyl chain unsaturation on MS peak intensity was minimized when the injection amount was maintained at less than 1 pmol. The method was applied for the quantitative calculation of 34 PC species from rat liver, which were identified from data‐dependent MS/MS analysis during nanoflow LC separation.