A RAPID AND PRECISE METHOD FOR THE DETER-MINATION OF VITAMIN D3 IN RAT SKIN BY NIGH-PERFORMANCE LIQUID CHROMATOGRAPHY
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概要
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In order to develop the investigations into photobiogenesis of vitamin D3, a rapid and precise method for the determination of the vitamin in rat skin was established by using high-performance liquid chromatography (HPLC). The proposed method included saponification of small pieces of rat skin, extraction of the unsaponifiable matter and application to HPLC using “Zorbax SIL” (straight-phase) as an adsorbent and 0.5% isopropanol in n-hexane as a mobile phase. The applicable lower limit of the method was 2ng of vitamin D3/cm2 of subcutaneous tissue-removed skin and it was possible to assay a concentration higher than 2ng/cm2. The proposed method was applied to determine the content of vitamin D3 in rat skin obtained from in vivo and in vitro irradiation experiments. In the in vitro experiment, the yield of vitamin D3 increased in proportion to the irradiation time. On the other hand, the yield in the in vivo experiment showed a proportional increase similar to the in vitro experiment until 60min irradiation, while a nearly constant value was obtained by irradiation for longer than 60min. When the rat skin obtained from the in vitro experiment was irradiated with monochromatic UV rays in the range 260-350nm, the most effective wavelength for the formation of vitamin D3 was confirmed to be 303nm, which differs from the result obtained from the experiment in a test tube (295nm). Moreover, the yield of vitamin D3 by irradiation with UV rays below 288nm was extremely low, which again differed from the results of a test tube experiment. These differences were thought to be due to the filter effect of the malpighian layer in the epidermis of rat skin.
著者
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小林 正
Department Of Biochemistry Kobe Gakuin University
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岡野 登志夫
Department of Hygienic Sciences, Kobe Women's College of Pharmacy
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高田 勝次
神戸女子薬大
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高田 勝次
Department of Hygienic Chemistry, Kobe Womens College of Pharmacy
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田村 葉子
Department of Hygienic Chemistry, Kobe Womens College of Pharmacy
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松井 幸子
Department of Hygienic Chemistry, Kobe Womens College of Pharmacy
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岡野 登志夫
Department of Hygienic Chemistry, Kobe Women's College of Pharmacy
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高田 勝次
Department of Hygienic Chemistry, Kobe Women's College of Pharmacy
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小林 正
Department of Hygienic Sciences, Kobe Women's College of Pharmacy
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小林 正
Department of Hygienic Scfences, Kobe Women's College of Pharmacy
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田村 葉子
Department of Hygienic Chemistry, Kobe Women's College of Pharmacy
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松井 幸子
Department of Hygienic Chemistry, Kobe Women's College of Pharmacy
関連論文
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