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Jones oxidation and high performance liquid chromatographic analysis of cholesterol in biological samples. 2007
Jun Dong, and
Wenxiang Chen, and
Shu Wang, and
Jiangtao Zhang, and
Hongxia Li, and
Hanbang Guo, and
Yong Man, and
Baosheng Chen
Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing 100710, China.
A simple pre-column derivatization procedure for HPLC analysis of cholesterol in biological samples was developed. Cholesterol was treated with chromic acid and sulfuric acid in acetone (the Jones oxidation) and cholest-4-en-3,6-dione was formed. The reaction was finished in 5 min at room temperature and the product showed a strong UV absorbance at 250 nm that enabled an HPLC detection limit of 0.2 pmol. With stigmasterol as an internal standard, the reaction was applied to the analysis of total and free cholesterol in serum and high-density lipoproteins and the analysis showed a within-run and total coefficient of variation of about 0.2% and 0.5%, respectively.
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