Isolation and purification of in vivo-generated 25-hydroxyvitamin D2 by preparative high-performance liquid chromatography.
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概要
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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.
- 財団法人 学会誌刊行センターの論文
著者
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小林 正
Department Of Biochemistry Kobe Gakuin University
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松尾 保
Department of Pediatrics Kobe University School of Medicine
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黒田 英造
Department of Pediatrics, Kobe University School of Medicine
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児玉 荘一
Department of Pediatrics, Kobe University School of Medicine
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岡野 登志夫
Department of Hygienic Chemistry, Kobe Women's College of Pharmacy
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松山 直子
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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