Determination of serum levels of unesterified lathosterol by isotope dilution-mass spectrometry

Scandinavian Journal of Clinical and Laboratory Investigation
E LundI Björkhem

Abstract

The synthesis of 2H3-labelled lathosterol is described. This compound was used together with 2H7-labelled cholesterol for simultaneous assay of unesterified lathosterol and cholesterol in serum by isotope dilution-mass spectrometry. After addition of a fixed amount of the two internal standards to a fixed amount of serum (in general 25 microliter), the steroids were extracted with chloroform and subjected to Lipidex 5000 chromatography. The fraction containing cholesterol and lathosterol was converted into trimethylsilyl ether and subjected to mass spectrometric analysis with selected monitoring of the ions at m/z 458 (molecular ion of the trimethylsilyl ether derivative of unlabelled cholesterol and lathosterol), m/z 461 (molecular ion of derivative of 2H3-labelled lathosterol) and m/z 465 (molecular ion of derivative of 2H7-labelled cholesterol). Individual standard curves were used for assay of each steroid. Under the conditions employed, the coefficient of variation of the two assays was less than 6%. In different recovery experiments the maximal difference between expected and found values was less than 7%. Using a less accurate method for analysis of lathosterol, we have shown previously that there is a high correlation b...Continue Reading

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