| Literature DB >> 27218859 |
Changhong Yun1, Taijun Yin1, Katherine Shatzer1, Douglas G Burrin2, Liwei Cui2, Yifan Tu1, Ming Hu3.
Abstract
An LC-MS/MS method was developed and validated to determine 7α-OH cholesterol in liver microsome. This method was convenient and fast with high specificity and sensitivity. Briefly, a gradient elution was performed on a Synergi polar-C18 column (50×4.6mm i.d., 3μm). The mobile phase (consisting of 0.1% HCOOH solution and acetonitrile) eluted in gradient at a flow rate of 1ml/min. MS detection was operated on APCI (+) mode; the MRM transitions for 7α-OH cholesterol and D7-cholesterol (I.S.) were 385.1≥159.1 and 376.4≥266.3, respectively. The linear response range of 7α-OH cholesterol was covered from 1.563 to 100.0ng/ml. All of the validation items meet the requirement of FDA guidance for bioanalytical method validation. This method was applied to enzymatic studies for determination of cholesterol 7alpha-hydroxylation activity catalyzed by CYP7A1 in the cholestatic minipigs liver microsomes.Entities:
Keywords: 7α-OH cholesterol; CYP7A1; Cholesterol 7α-hydroxylase activity; LC–MS/MS; Liver microsome
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Year: 2016 PMID： 27218859 PMCID： PMC5358015 DOI： 10.1016/j.jchromb.2016.05.005
Source DB: PubMed Journal: J Chromatogr B Analyt Technol Biomed Life Sci ISSN： 1570-0232 Impact factor: 3.205