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  • 标题:IDENTIFICATION AND DETERMINATION OF 7-DEHYDROCHOLESTEROL IN RAT SKIN
  • 本地全文:下载
  • 作者:Mitsue YASUMURA ; Toshio OKANO ; Kumiko MIZUNO
  • 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:1977
  • 卷号:23
  • 期号:6
  • 页码:513-523
  • DOI:10.3177/jnsv.23.513
  • 出版社:Center for Academic Publications Japan
  • 摘要:Lipids were extracted from the skin of normal rats with chloroform-methanol (1:1) and the solvent evaporated. The resulting residue was saponified, and the unsaponifiable matter extracted with benzene was purified by subjecting the extract successively to hydroxy-alkoxypropyl (HAP) Sephadex column chromatography, silica-gel column chromatography and preparative thin-layer chromatography (TLC). The resulting purified provitamin D fraction was confirmed to contain 7-dehydrocholesterol (7-DHC) based on data of ultraviolet (UV) spectrum, high-performance liquid chromatography (HPLC), gas-liquid chromatography (GLC) and gas-liquid chromatography-mass spectro-metry (GC-MS). Based on the results of GC-MS and mass chromato-graphy, it was also confirmed that the fraction contained small amounts of cholesterol, lathosterol and two other unknown sterols besides 7-DHC. Then, the determination of 7-DHC in rat skin was investigated and a HPLC method was proposed. This method included extraction of lipids from small pieces of rat skin with chloroform-methanol (1:1), saponifica-tion, isolation of the unsaponifiable matter and application to HPLC. The estimated average values of 7-DHC in normal rat skin (20 weeks old), with and without UV irradiation, were 107.7 and 103.8μg/g, respectively. These were significantly higher than the corresponding values of vitamin D-deficient rat skin. UV irradiation of rat skin did not affect the change of 7-DHC content in the tissue.
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