Lab 3 ALT and AST Daheeya Al Enazi

Lab 3 ALT and AST Daheeya Al. Enazi Cromatest Kit

Quantitative Determination of Alanine aminotransferase (ALT) q. Test is typically used to detect liver injury. q. It is often ordered in conjunction with aspartate aminotransferase (AST) or as part of a liver panel to screen for and/or help diagnose liver disease. q Although ALT is more specific to the liver than is AST. q. ALT is often used to monitor the treatment of persons who have liver disease

Signs and Symptoms of liver disease: ü • Weakness, fatigue ü • Loss of appetite ü • Nausea, vomiting ü • Abdominal swelling and/or pain ü • Jaundice ü • Dark urine, light-colored stool

Principle L-Alanine + 2 Oxoglutarate ALT L- Glutamate + Pyruvate + NADH + H+ LDH Lactate + NAD+ +H 2 O The rate of decrease in NADH absorbance at 340 nm is proportional to the ALT activity of the sample.

Clinical significance q. Hepatic disorders, high serum levels are found in hepatocellular disorder rather than on obstructive disorders. q. Cardiac tissue contains a small amount of ALT activity but serum levels usually remain normal in AMI. q. Test used for infectious and toxic hepatitis

procedure WR-ml (mix 4 ml of 1 R 1+1 ml f R 2) Sample-µl 50 ØMix, incubate at 37℃ for 1 minute and read the absorbance at 340 nm against D. W, and after 3 min read the absorbance Ø Calculation: ΔA/min x 3333= U/L of ALT ØNormal Range: (ALT: at 37 C) Adult up to 40 U/L

Quantitative Determination of Aspartate aminotransferase (AST) L-Aspartate +2 Oxoglutarate AST > L- Glutamate+ Oxaloacetate + NADH +H+ MDH > L-Malate + NAD+ The rate of decrease of NADH absorbance at 340 nm is proportional to the AST activity of the sample.

Clinical significance ØThe AST found in highest conc. in heart muscle, liver and skeletal muscle. ØSmaller amounts are found in kidney, pancreas and erythrocytes. ØClinical use: Ømyocardial infarction, hepatocellular disease, and disorders of skeletal muscles.

procedure WR-ml(4 ml R 1+1 ml R 2) 1 Sample -µl 50 Mix, incubate at 37℃ for 1 minute. Read initial absorbance of the sample, start the stopwatch and read Abs. for 3 minutes. Calculate the difference between absorbance and the average absorbance differences per minute ( ΔA/min)

Calculations ΔA/min x 3333= U/L of AST ØReference values: (at 37 C) Adult up to 40 U/L