A general colorimetric procedure for the estimation of enzymes which are linked to the NADH/NAD+ system

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Abstract

  • 1.

    I. Two colour reactions for determining the reduced coenzyme NADH are described.

    • 1.1.

      (a) A coloured formazan is produced by the reduction of the tetrazolium salt INT with NADH, electron transfer being facilitated by PMS ; the formazan is stable and the extinction is read at 500 mμ.

    • 1.2.

      (b) The coloured ferrous dipyridyl complex is produced following the reduction of a ferric salt with NADH, in the presence of PMS; the coloured complex is stable for 30 min. The extinction is read at 520 mμ.

    I μmole of NADH, when reacted with the ferric salt, gives the same absorbance as 2 τmmoles of a ferrous salt, and it is possible to standardise the solution of NADH against a stable iron standard without the need for an ultraviolet spectrophotometer.

  • 2.

    2. Details are given of tlie assay of HBD, LDH, SDH, MDH and GOT, GPT, using these colour reactions, micro-volumes of sample being used.

  • 3.

    3. Details are given of standard deviation, cost, reliability, etc., of the methods described.

  • 4.

    4. Present methods for the assay of GOT and GPT are discussed in the light of recent literature.

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