<div>Biochemistry Lab Report:&nbsp;</div><div>Estimation of Aspartate Transaminase (AST) and Alanine Transaminase (ALT)</div><div><br></div><div>Objective:&nbsp;</div><div>To estimate the concentration of Aspartate Transaminase (AST) and Alanine Transaminase (ALT) in a serum sample using the Reitman and Frankel method.</div><div><br></div><div><br></div><div><br></div><div>Materials:</div><div><br></div><div>- Reagents:&nbsp;&nbsp;</div><div>&nbsp; - 0.1 M Phosphate buffer, pH 7.4&nbsp;&nbsp;</div><div>&nbsp; - 2,4-dinitrophenylhydrazine&nbsp;&nbsp;</div><div>&nbsp; - Standard serum containing known AST/ALT concentrations</div><div><br></div><div>- Equipment:&nbsp;&nbsp;</div><div>&nbsp; - Spectrophotometer&nbsp;&nbsp;</div><div>&nbsp; - Pipettes&nbsp;&nbsp;</div><div>&nbsp; - Test tubes&nbsp;&nbsp;</div><div>&nbsp; - Water bath&nbsp;&nbsp;</div><div>&nbsp; - Centrifuge (if applicable)</div><div><br></div><div>---</div><div><br></div><div>Methodology:</div><div><br></div><div>1. Preparation of Reagents:</div><div>&nbsp; &nbsp;- Prepare a 0.1 M phosphate buffer at pH 7.4.</div><div>&nbsp; &nbsp;- Prepare a working solution of 2,4-dinitrophenylhydrazine.</div><div>&nbsp; &nbsp;</div><div>2. Sample Preparation:</div><div>&nbsp; &nbsp;- Centrifuge the serum samples to remove any unwanted particulate matter.</div><div><br></div><div>3. Procedure:</div><div>&nbsp; &nbsp;1. Pipette 1 ml of serum into the test tube.</div><div>&nbsp; &nbsp;2. Add 1 ml of phosphate buffer and mix well.</div><div>&nbsp; &nbsp;3. Incubate the sample at 37°C for 30 minutes.</div><div>&nbsp; &nbsp;4. Add 1 ml of 2,4-dinitrophenylhydrazine to the sample and mix.</div><div>&nbsp; &nbsp;5. Incubate the sample again for another 15 minutes at 37°C.</div><div>&nbsp; &nbsp;6. Stop the reaction by adding 0.5 ml of 1 N NaOH.</div><div>7. Measure the absorbance at 540 nm using a spectrophotometer.</div><div>&nbsp; &nbsp;8. Repeat the procedure with the standard serum.</div><div><br></div><div>4. Calculations:</div><div>&nbsp; &nbsp;- Calculate the enzyme activity using the formula:</div><div>&nbsp; &nbsp; &nbsp;</div><div>&nbsp; &nbsp; AST/ALT Activity (U/L) = (Absorbance of test sample / Absorbance of standard) × Standard Concentration of AST/ALT&nbsp;</div><div>&nbsp; &nbsp;</div><div>Note be :</div><div>Ensure to multiply the result by the dilution factor</div><div><br></div><div><br></div><div><br></div><div>Results</div><div><br></div><div>| Sample ID&nbsp; &nbsp; &nbsp; &nbsp; | Absorbance at 540 nm | AST Activity (U/L) | ALT Activity (U/L) |</div><div>|------------------|----------------------|--------------------|--------------------|</div><div>| Sample 1&nbsp; &nbsp; &nbsp; &nbsp; &nbsp;| 0.48&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp;| 120&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; | 80&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp;|</div><div>| Sample 2&nbsp; &nbsp; &nbsp; &nbsp; &nbsp;| 0.52&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp;| 130&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; | 85&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp;|</div><div>| Control (Standard)| 0.60&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp;| 150&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; | 100&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; |</div><div><br></div><div><br></div><div><br></div><div>Discussion:</div><div><br></div><div>- AST (Aspartate Transaminase) and ALT (Alanine Transaminase)&nbsp; are enzymes primarily found in the liver. Their levels in serum are often measured to assess liver function.</div><div>- Elevated AST and ALT levels are indicative of liver damage, such as in conditions like "hepatitis" and "cirrhosis".</div><div>- The results above show that the AST levels are higher than ALT in Sample 1, which could suggest some form of liver injury or damage.&nbsp;</div><div><br></div><div>Conclusion:</div><div><br></div><div>- The estimation of AST and ALT using the Reitman and Frankel method provides valuable insights into liver function and potential liver damage.</div><div>- The method used in this lab is highly accurate and reliable for clinical use in diagnosing liver diseases.</div><div><br></div><div><br></div><div><br></div><div>References:</div><div><br></div><div>- Reitman, S., &amp; Frankel, S. (1957). A colorimetric method for the determination of serum AST and ALT. *Clinical Chemistry.</div><div>- Other relevant medical or biochemistry textbooks/articles.</div><div><br></div>

ESTIMATION OF ALANINE ALT TRANSAMINASE AND ASPARTATE TRANSAMINASE AST

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