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Method to determine 4‐oxo‐retinoic acids, retinoic acids and retinol in serum and cell extracts by liquid chromatography/diode‐array detection atmospheric pressure chemical ionisation tandem mass spectrometry
Author(s) -
Rühl Ralph
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.2621
Subject(s) - retinoic acid , chemistry , chromatography , retinol , high performance liquid chromatography , tandem mass spectrometry , mass spectrometry , liquid chromatography–mass spectrometry , biochemistry , vitamin , gene
Retinoic acid isomers, 4‐oxo‐retinoic acid isomers and retinol are present in the serum of mammals. In this study a high‐performance liquid chromatography (HPLC) separation, sample preparation and tandem mass spectrometry (MS/MS) method was established for quick and easy sample preparation and sensitive determination of retinoids such as all‐ trans ‐4‐oxo‐retinoic acid, 13‐ cis ‐4‐oxo‐retinoic acid, 13‐ cis ‐retinoic acid, 9‐ cis ‐retinoic acid, all‐ trans ‐retinoic acid and retinol in serum and cell extracts. Serum samples were simply treated with three times the volume of isopropanol, dried under vacuum, taken up in the HPLC solvent and immediately put into the autosampler for an automated single‐run HPLC analysis. With this MS/MS method we were able to detect 7 pg and quantify 20 pg of all‐ trans ‐retinoic acid, 4‐oxo‐all‐ trans ‐retinoic acid and retinol directly on‐column and were able to determine a concentration as low as 0.2 ng/mL in ethanolic standards and in biological samples. This method allows ultra‐sensitive detection, excellent selectivity and a very simple sample preparation to determine retinoic acids, 4‐oxo‐retinoic acids and retinol in serum and cell extracts for the study of endogenous retinoids. Copyright © 2006 John Wiley & Sons, Ltd.