Determination of α-Tocopherol and α-Tocopherylquinone in Rat Tissues and Plasma by High-Performance Liquid Chromatography with Electrochemical Detection
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概要
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α-Tocopherol and α-tocopherylquinone in rat tissues and plasma were determined simulaneously by using high-performance liquid chromatography-electrochemical detection (HPLC-ED) with dual electrodes in the series mode. Biological samples were saponified in the presence of a mixture of butylated hydroxytoluene, ascorbic acid, and pyrogallol and then extracted with hexane. The compounds were separated on a C18 column using a mobile phase containing 95% methanol and 0.05 M sodium perchlorate as the supporting electrolyte. After HPLC separation, α-tocopherylquinone was first reduced at an upstream electrode at -500mV. Both α-tocopherol and the reduction product of α-tocopherylquinone were then oxidized downstream at +600 mV. Only the downstream electrode current was monitored for the determination. Linearity of the standard curves was obtained over the range 5-30 pmol for α-tocopherol and α-tocopherylquinone. Minimum detectable quantities(S/N of 3) were 0.25 pmol for α-tocopherol and 0.31 pmol for α-tocopherylquinone. The method was applied to analysis of the contents of α-tocopherol and α-tocopherylquinone in rat tissues and plasma. By hyperoxia, the content of α-tocopherol was decreased remarkably in lung, and in contrast, the contents of α-tocopherylquinone were increased in all tissues studies with the exception of plasma, though the content of α-tocopherylquinone in normal rats is quite small. The technique is particularly useful in the quantitation of the oxidation of α-tocopherol in biological samples.
- 公益社団法人日本薬学会の論文
- 2000-10-01
著者
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金澤 秀子
共立薬科大学
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金澤 秀子
Faculty Of Pharmacy Keio University
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Urano Shiro
Shibaura Institute Of Technology
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MATSUSHIMA Yoshikazu
Kyoritsu College of Pharmacy
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KANAZAWA Hideko
Kyoritsu University of Pharmacy
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MIYATA Chizuko
Kyoritsu College of Pharmacy
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NAGATA Yoshiko
Kyoritsu College of Pharmacy
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Matsuoka Yuji
School Of Medicine Fukuoka University
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Nagata Y
Kyoritsu College Of Pharmacy
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Matsushima Y
Kyoritsu College Of Pharmacy
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Kanazawa H
Dept. Biotechnology Faculty Of Engineering Sciences
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金澤 治男
Kyoritsu College Of Pharmacy
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Kanazawa Hideko
Kyoritsu College Of Pharmacy
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