An Improved Method for Determination of Ascorbic Acid in Blood Plasma and Urine by α,α'-Dipyridyl Method

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  • OKAMURA Masato
    Laboratory of Biochemistry,Kanagawa Prefectural College of Nursing and Medical Technology

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Other Title
  • α,α'-Dipyridyl法による血漿及び尿中のアスコルビン酸の定量法
  • α,α′-Dipyridyl法による血漿及び尿中のアスコルビン酸の定量法
  • アルファ アルファ Dipyridylホウ ニヨル ケッショウ オヨビ ニョウ

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Description

Various conditions for reducing dehydroascorbic acid (DAsA) were studied, using dithiothreitol (DTT). This led to an improved method for the separate determination of ascorbic acid (AsA) and DAsA in blood plasma, and the specific method for determination of total AsA in urine. (1) Separate determination of AsA and DAsA in blood plasma. The content of AsA was determined by α,α'-dipyridyl method (A). DAsA was reduced to AsA by incubation with DTT. After removing the excess DTT with N-ethylmaleimide (NEM), total AsA, i.e. reduced plus original, were determined by theα,α'-dipyridyl method (B). The amount of DAsA could then be calculated from these results (B minus A). (2) Determination of total AsA in urine. The urine sample was acidified with trichloroacetic acid and was shaken with activated carbon to remove interfering substances. As AsA was oxidized with this treatment, the resultant DAsA was reduced by the DTT-NEM system for determination byα,α'-dipyridyl method.

Journal

  • VITAMINS

    VITAMINS 55 (11), 495-508, 1981

    THE VITAMIN SOCIETY OF JAPAN

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