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Quantitative Determenation of Alanine aminotransferase (ALT)

Quantitative Determenation of Alanine aminotransferase (ALT) . CLS 431. Determenation of Alanine aminotransferase (ALT). Principle of method Alanine + 2-oxoglutrate ALT Glutamate + Pyruvate Pyruvate + NADH + H + LDH Lactate + NAD +

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Quantitative Determenation of Alanine aminotransferase (ALT)

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  1. Quantitative Determenation of Alanineaminotransferase (ALT) CLS 431

  2. Determenation of Alanine aminotransferase (ALT) • Principle of method • Alanine + 2-oxoglutrate ALT Glutamate + Pyruvate • Pyruvate + NADH + H+LDH Lactate + NAD+ The rate of decrease in absorbance at 340nm is proportional to the ALT activity of the sample.

  3. Enzyme distribution: Found in highest conc. in liver and kidney. It is considered to be the most liver specific transaminase

  4. Clinical significance of ALT • hepatic disorders, hight serum levels are found in hepatocellular disorder rather than on obstructive disorders. • Cardiac tissue contains a small amount of ALT activity but serum levels usually remain normal in AMI.

  5. Samples:serum or plasma • Procedure • Assay conditions: • Wavelength ......................... 340nm. • Cuvette ................................ 1cm. light path • constant tempreture .................. 37℃ • Adjust the instrument to zero with distilled water or air.

  6. Pipette into a cuvette: • Mix, incubate at 37℃ for 1 minute and read the absorbance at 340 nm against D.W. (A1). • Take the Ab every minutes for 3 min read the absorbance(A2).

  7. Calculations ΔA/min x 1768= U/L of AST • Reference values: • ALT: • Men up to 42 U/L • Women up to 32 U/L

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