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Journal of Lipid Research, Vol 18, 232-238, Copyright © 1977 by Lipid Research, Inc.

ARTICLES

Bile acid synthesis in man. II. Determination of 7 alpha- hydroxycholesterol, (22R)-22-hydroxycholesterol, and 26- hydroxycholesterol in human meconium

U Lavy, S Burstein, M Gut and NB Javitt

7 alpha-Hydroxycholesterol, (22R)-22-hydroxycholesterol and 26- hydroxycholesterol have been quantitated in human meconium. The method used tetrahydrofuran for extraction and solvolysis of the sulfate esters, liquid partition chromatography for the separation of the hydroxysterols, gas-liquid chromatography for quantitation, gas-liquid chromatography-mass spectrometry for identification, and tritiated and 14C-labeled tracers for overall recovery standards. (22R)-22- Hydroxycholesterol and 26-hydroxycholesterol were present almost entirely, ( greater than 93%) in the sulfate fraction at concentrations of 3.8-6.4 and 0.4-0.8 mg per 100 g meconium, respectively. Since free tritiated (22R)-22-hydroxycholesterol was used as the tracer to assess recovery of this hydroxysterol, the concentrations found for this compound may be minimal. Tritiated 26-hydroxycholesterol 3,26-disulfate was used as tracer to determine the levels of this compound, and the solvolysis procedure was optimized for recovery of 26- hydroxycholesterol and least decomposition of 7 alpha- hydroxycholesterol. No significant amounts of 7 alpha- hydroxycholesterol were found based on the tracer-free hydroxysterol as recovery standard.
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