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High performance liquid chromatography assay for alanine amino transferase

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Summary

An improved method of alanine-amino transferase determination is proposed. The reaction is carried out with alanine and 2-oxoglutarate as substrates and analysis is by HPLC on a reversed-phase chromatographic system using a C18 column and tetrabutylammonium phosphate in phosphate buffer (pH 7.0)-acetonitrile as mobile phase. The enzyme activity was determined by directly following the formation of pyruvic acid without employing any secondary reaction, which is necessary in the spectrophotometric method. The detection limit of pyruvic acid is 10 pmole μl−1 and the standard deviation for the enzymatic activity of standard solutions is 5.4%. Furthermore under the chromatographic conditions selected it is possible to detect the presence of some intermediate species.

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Authors and Affiliations

  1. Dipartimento di Chimica, Università di Roma “La Sapienza”, P.le Aldo Moro 5, 00185, Roma, Italy

    S. Canepari, V. Carunchio, A. M. Girelli & A. Messina

Authors
  1. S. Canepari
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  2. V. Carunchio
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  3. A. M. Girelli
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  4. A. Messina
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Additional information

Work supported by National Research Council of Italy. Presented in part at the “First International Symposium on Applications of HPLC in Enzyme Chemistry”, Verona, September 1990.

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Canepari, S., Carunchio, V., Girelli, A.M. et al. High performance liquid chromatography assay for alanine amino transferase. Chromatographia 32, 23–26 (1991). https://doi.org/10.1007/BF02262461

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  • DOI: https://doi.org/10.1007/BF02262461

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