FACULTY OF SCIENCE

BACHELOR OF SCIENCE (HONS) BIOCHEMISTRY

UDEE3144 CLINICAL BIOCHEMISTRY

PRACTICAL REPORT 1

Name : Poon Wei Chi

Student ID : 1805087
Practical group : P1
Lecturer’s name : Dr. Anto Cordelia Vijanth
Title
Estimation of transaminase

Aim
To understand the principle and determine SGOT (AST) activity in given serum sample

Principle

 SGOT catalyses the transfer of Amino group from L-Aspartate to α-ketoglutarate

resulting in formation of oxaloacetate and glutamate

 Oxaloacetate formed reacts with 2,4-dinitrophenyl hydrazine to produce a hydrazone

derivative, which in alkaline medium produces a brown colored complex whose intensity
is measured colorimetrically at 520 nm

Reagents and equipment

 Small test tubes, micropipettes, constant temperature water bath, spectrophotometer

 Substrate buffer pH 7.4, 2,4-dinitrophenylhydrazine, 200 mM α-ketoglutarate, 0.4 M
NaOH

Procedures

i) Preparation for blank and test sample

1) 0.5 mL substate and 0.1 mL serum were mixed in a test tube.
2) 0.5 mL substrate and 0.1 mL sterile water were mixed in a test tube.
3) Two test tubes were incubated at 37°C for 30 minutes
 4) 0.5 mL 2,4-DNPH was added into the two test tubes respectively and sited for 20
minutes at room temperature.
5) 5.0 mL of 0.4 M NaOH was added into two test tubes respectively and sited for
10 minutes.
6) The absorbance for two test tubes were read at 520 nm.
ii) Preparation of standard curve
1) Six test tubes were prepared according to the following table:

Test tube 1 2 3 4 5 6
Substrate
0.5 0.4 0.3 0.2 0.1 0.05
(mL)
Std OAA
(2 µmol/mL) - 0.1 0.2 0.3 0.4 0.45
(mL)

2) Six test tubes were incubated at 37°C for 30 minutes.

3) 0.5 mL 2,4-DNPH was added into the six test tubes respectively and sited for 20
minutes at room temperature.
4) 5 mL of 0.4 M NaOH was added into the six test tubes respectively and sited for
10 minutes.
5) The absorbance for six test tubes were read at 520 nm.

Discussion

SGOT test is used to determine the level of the SGOT which is the tissue enzyme catalyse the
exchange of amino and keto groups between alpha-amino acid and alpha-keto acid. This test
could help doctor in diagnosis of the liver disease, heart disease, kidney disease or muscle
damage through the blood test (Gotter, 2018). The SGOT is found in all tissues including liver,
heart and kidney. But the tissue contains the highest of the SGOT activity is in heart or cardiac
muscle. If there are damage of liver, heart, kidney or muscle the SGOT level will be higher than
normal. The normal level of the SGOT level is lower than 36 U/L (Fletcher and Sissons, 2021).
Normally, the patient with the high level of SGOT level may has myocardial infarction or
hepatitis. While the low than normal level of SGOT may face with vitamin B6 deficiency, liver
or kidney disease.

Conclusion

SGOT test is one of the diagnoses for certain disease such as heart, liver or kidney disease via
measure the amount of the SGOT presence in blood.

References

Gotter, A., 2018. SGOT Test. Available at:< https://www.healthline.com/health/sgot-

test#followup> [Accessed 25 February 2022]

Fletcher, J. and Sissons, B., 2021. AST (SGOT) blood test result ranges, and how to interpret
them. Available at:< https://www.medicalnewstoday.com/articles/320982> [Accessed 25
February 2022]