Determination of α-tocopherol and α-tocopherylquinone in rat tissues and plasma by high-performance liquid chromatography with electrochemical detection

Hideko Kanazawa, Chizuko Miyata, Yoshiko Itoh, Shiro Urano, Yoshikazu Matsushima

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

α-Tocopherol and α-tocopherylquinone in rat tissues and plasma were determined simultaneously 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 -500 mV. 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 studied 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.

Original languageEnglish
Pages (from-to)1462-1466
Number of pages5
JournalChemical and Pharmaceutical Bulletin
Volume48
Issue number10
Publication statusPublished - 2000

Fingerprint

Tocopherols
High performance liquid chromatography
Rats
High Pressure Liquid Chromatography
Tissue
Plasmas
Electrodes
Pyrogallol
Butylated Hydroxytoluene
Hyperoxia
tocopherylquinone
Hexanes
Electrolytes
Ascorbic Acid
Methanol
Oxidation
Lung

Keywords

  • α-Tocopherol
  • α-Tocopherylquinone
  • HPLC-ED
  • Hyperoxia
  • Rat tissue

ASJC Scopus subject areas

  • Drug Discovery
  • Organic Chemistry
  • Chemistry(all)
  • Pharmacology

Cite this

Determination of α-tocopherol and α-tocopherylquinone in rat tissues and plasma by high-performance liquid chromatography with electrochemical detection. / Kanazawa, Hideko; Miyata, Chizuko; Itoh, Yoshiko; Urano, Shiro; Matsushima, Yoshikazu.

In: Chemical and Pharmaceutical Bulletin, Vol. 48, No. 10, 2000, p. 1462-1466.

Research output: Contribution to journalArticle

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