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A new approach to overcome natural cholesterol interference during simultaneous determination of two stable isotope‐enriched cholesterol tracers in human plasma
Author(s) -
Wolff E.,
Vergnes M. F.,
Kaloustian J.,
Abou L.,
Mikail C.,
Lairon D.,
Portugal H.,
Nicolay A.
Publication year - 2007
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.3193
Subject(s) - chemistry , chromatography , isotopomers , analyte , reagent , repeatability , isotope , interference (communication) , reproducibility , cholesterol , mass spectrometry , tracer , analytical chemistry (journal) , gas chromatography , molecule , biochemistry , organic chemistry , quantum mechanics , nuclear physics , electrical engineering , engineering , channel (broadcasting) , physics
We have developed a validated gas chromatography/mass spectrometry (GC/MS) method with two labelled cholesterol tracers, i.e. 2 H 4 ([2H4]‐Chol) and 2 H 7 ([2H7]‐Chol) enriched moieties, with a new way of calculating the abundance of labelled cholesterol in plasma without natural cholesterol interference. The isotopomers of the analytes could interfere during analysis. Elimination of these interferences can be performed by the blank or mathematical subtraction method. Validation was performed with the two interference elimination methods. For both methods, linearity was obtained in the range 5 × 10 −4 to 10 −2 mM for both labelled cholesterol moieties. In the same range, repeatability and reproducibility were less than 6.5% and 7.5% for [2H4]‐Chol and [2H7]‐Chol, respectively. Accuracy was about 100% and recoveries always included 100% for the two labelled cholesterols. We demonstrate that measurement of blank plasma is not necessary when using the validated abundance isotope calculation method. This saves time, reagent and samples. This calculation strategy can be extrapolated to comparable tracer approaches. Copyright © 2007 John Wiley & Sons, Ltd.