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  • Title: Measurement of cholesterol 7 alpha-hydroxylase activity with selected ion monitoring.
    Author: Sanghvi A, Grassi E, Bartman C, Lester R, Galli Kienle M, Galli G.
    Journal: J Lipid Res; 1981 May; 22(4):720-4. PubMed ID: 7276747.
    Abstract:
    Formation of 7 alpha-hydroxycholesterol by rat liver microsomes was quantitated using a gas chromatograph-mass spectrometer operated in selected ion monitoring mode. This procedure measures the mass of 7 alpha-hydroxycholesterol formed and does not require addition of labeled cholesterol to monitor the reaction, since endogenous microsomal cholesterol serves as substrate for the enzyme cholesterol 7 alpha-hydroxylase. The method requires only the extraction of lipids from microsomal incubations and injection of a small fraction of this extract into the apparatus. Microsomes from normal rat livers incubated for different periods were found to yield increased 7 alpha-hydroxycholesterol with time.
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