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High‐performance liquid chromatography/atmospheric pressure chemical ionization tandem mass spectrometry determination of cholesterol uptake by Caco‐2 cells
Author(s) -
Tian Qingguo,
Failla Mark L.,
Bohn Torsten,
Schwartz Steven J.
Publication year - 2006
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.2700
Subject(s) - chemistry , atmospheric pressure chemical ionization , chromatography , tandem mass spectrometry , liquid chromatography–mass spectrometry , mass spectrometry , cholesterol , high performance liquid chromatography , chemical ionization , methanol , analytical chemistry (journal) , ionization , ion , biochemistry , organic chemistry
A simple, sensitive and selective liquid chromatography/atmospheric pressure chemical ionization tandem mass spectrometric method (LC/APCI‐MS/MS) was developed and applied to quantitative determination of uptake of cholesterol by Caco‐2 human intestine cells. Caco‐2 cells were cultured in medium containing cholesterol‐3,4‐ 13 C 2 and phytosterols from nutritional supplements after in vitro digestion. Cellular cholesterol (cholesterol‐3,4‐ 13 C 2 ) and endogenous cholesterol were extracted using methanol/chloroform (1:2, v/v) and directly analyzed using LC/APCI‐MS/MS with selected reaction monitoring (SRM), using cholesterol‐2,2,3,4,4,6‐ d 6 as an internal standard. Detection and quantification limits were 2.2 and 7.2 pmol, respectively. This method provides an effective tool for rapid determination of cholesterol uptake by cells with increased selectivity and sensitivity in comparison to previously reported LC/APCI‐MS analysis using selected ion monitoring (SIM). Copyright © 2006 John Wiley & Sons, Ltd.