期刊名称:Journal of Nutritional Science and Vitaminology
印刷版ISSN:0301-4800
电子版ISSN:1881-7742
出版年度:1979
卷号:25
期号:6
页码:479-493
DOI:10.3177/jnsv.25.479
出版社:Center for Academic Publications Japan
摘要:In order to obtain a standard compound of 25-hy-droxyvitamin D2 (25-OH-D2), a method for isolating in vivo -generated 25-OH-D2 from the blood of rats or rabbits was established by using several steps of preparative high-performance liquid chromatography (HPLC). When the unsaponifiable matter of the plasma obtained from rats or rabbits receiving a large dose of vitamin D2 was applied to the preparative HPLC using a Zorbax SIL column, a peak denoted as peak X was observed on the chromatogram. Since the peak X was thought to be due to 25-OH-D2 from the experiments of time course and dose-response, it was purified by subjecting it to successive preparative HPLC using several kinds of columns. From the results of ultraviolet (UV) absorption spectrum, gas chromatography-mass spectrometry (GC-MS) and mass chromatography, the purified peak X compound was confirmed to be 25-OH-D2. The proposed method for isolating in vivo -generated 25-OH-D2 is very convenient, because the time to perform each HPLC is very short though several steps of HPLC are used.