Cagayan State University - Andrews Campus, Tuguegarao City

CLINICAL CHEMISTRT 2 Laboratory | TTh, 5:00 PM - 8:00 PM

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Wilson Leal Laranang, RMT, MSc Not started

GROUP 6 Date Submitted: 10/07/2022

GALANG, OLAND DEEMS B.
VILLANUEVA, KREZA LYN E.
VILLENA, JEIZEL CLAIRE G.

BSMLS-4 Rating: ________________

Activity No. 3A
ASPARTATE AMINO TRANSFERASE (AST) DETERMINATION

1. Describe the principle of aspartate amino transferase kinetic method of determination.

With the help of the enzyme aspartate aminotransferase (AST), oxaloacetate and L-
glutamate are produced when the amino group from L-aspartate is transferred to alpha-
ketoglutarate. The reduction of oxaloacetate and simultaneous oxidation of NADH+ to NAD are
catalyzed by malate dehydrogenase (MDH). As a result, the AST activity is directly proportional
to 340 nm which is the rate of decrease in absorbance.  To counteract endogenous pyruvate,
which is often found in serum, lactate dehydrogenase (LDH) is administered. (SGOT Test
Procedure, 2019, para 2; McPherson & Pincus, 2017, p. 295)

Figure 1. Aspartate aminotransferae (AST)/ Serum glutamic oxaloacetic transaminase (SGOT)

Note. AST catalyzes in the reaction (left) and

the assay system for measuring transaminase
activity (right). From Tietz Textbook of Clinical Chemistry and Molecular Diagnostics (6th ed., pp.
412, 414), by N. Rifai, et al. 2018. Elsevier Inc. Copyright 2018.
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CLINICAL CHEMISTRT 2 Laboratory | TTh, 5:00 PM - 8:00 PM
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2. What are the conditions affecting the AST determination?
TABLE 1. Conditions affecting the AST determination.
Elevated AST  Acute Myocardial Infarction
 Patients with chronic alcohol abuse
 Cirrhosis causing chronic hepatocyte injury
 Alcoholic hepatitis causing acute alcohol-induced hepatocyte
injury
Slight to moderate  Acute pancreatitis
elevation of AST  Crushed muscle injury
 Hemolytic disease
AST 2-3 x N Pulmonary emboli

AST 8 x N  Progressive muscular dystrophy

 Dermatomyositis
Elevated  The workup of patients with negative viral markers and negative
transaminases for drugs or alcohol ingestion should include less common
causes of hepatic injury:
o Hemochromatosis
o Wilson disease
o Autoimmune hepatitis
o Primary biliary cirrhosis
o Sclerosing cholangitis
o Celiac disease
o Alpha-1-antitrypsin deficiency
 Administration of various medications, such as:
o Non-steroidal anti-inflammatory drugs
o Antibiotics
o Antiepileptic drugs
o Statins, or opiates
o Over-the-counter medications
o Herbal preparations
 Primary or metastatic carcinoma of the liver
 Non-alcoholic fatty liver disease (NAFLD)
 Viral and alcoholic hepatitis
 Acute fulminant hepatic failure
Not elevated or low End-stage cirrhosis

Note. Adapted from Henry’s Clinical Diagnosis and Management by Laboratory Methods (23rd
ed. pp.295-296,301) by R.A. McPherson & M.R. Pincus, 2017. Copyright 2017 by Elsevier Inc;
Tietz Textbook of Clinical Chemistry and Molecular Diagnostics (6th ed. p.413) by N. Rifai et al.,
2018. Copyright 2018 by Elsevier Inc.
Cagayan State University - Andrews Campus, Tuguegarao City
CLINICAL CHEMISTRT 2 Laboratory | TTh, 5:00 PM - 8:00 PM
Instructor Status Notes

Wilson Leal Laranang, RMT, MSc Not started

3. What are the advantages of the kinetic method over the colorimetric method of AST
determination?
Colorimetric methods are inaccurate because dinitrophenylhydrazine reacts with a-
ketoglutarate and oxaloacetate, resulting in high reagent blanks (Colorimetric determination
of serum GOT, n.d., para 3) , and lack specificity because it reacts with any keto-compound
(Policarpio, 2021, p.39). In contrast, kinetic methods are specific in that they catalyze the
transfer of the amino group of aspartic acid to a-ketoglutarate (AST/GOT Liquiform, n.d., para
2).

4. What are the interfering factors in AST determination?

TABLE 2. Factors interfering in AST determination.

Due to differences in body mass, AST tends to be slightly higher in males than in females and
fluctuates with age. Up until the age of 15 to 20 years, AST is slightly greater than ALT. After
that, AST activity usually peaks later than ALT. AST and ALT activity roughly equalize at the
age of 60.

African males had AST levels that are 15% greater than those of Caucasian men.

Men who are obese may have slightly raised AST levels. The same person’s AST levels can
change by between 5 and 10% from day to day. For up to 24 hours, moderate exercise raises
AST levels, though this is typically shorter.

Although many medications have the potential to harm the liver, only few of users actually
experience this side effect, which can elevate AST. Natural products and supplements fit this
description as well.

Drugs and supplements that can increase AST include:

 NSAIDs like ibuprofen (Advil)
 Blood pressure medications
 Opiates
 Anti-seizure medications
 Chemotherapy agents
 Kava

Sex, age, race, diet, exercise, and the medications and supplements a person is taking are
just a few of the variables that can affect an AST result.

Note. Adapted from “Aspartate Aminotransferase” (para 2) by A. Cooper, 2019. Copyright 2022
by Exeter Clinical Laboratory International; High/Low Aspartate Aminotransferase
Levels+Function & Diseases (para 27) by P. Yazdi, 2021. Copyright 2013-2022 by SelfCode.
Cagayan State University - Andrews Campus, Tuguegarao City
CLINICAL CHEMISTRT 2 Laboratory | TTh, 5:00 PM - 8:00 PM
Instructor Status Notes

Wilson Leal Laranang, RMT, MSc Not started

5. What are the pre-analytical factors to consider when testing AST?

Wrong sample tube the sample is already contaminated and
should be rejected.
Patient with severe vitamin B6 deficiency

Hemolysis increases the concentration of aspartate

aminotransferase and can also interfere with
the reaction mechanism of AST testing (Irjala
& Grönroos, 1998).
Medication many different medicines and supplements
might alter overall AST level and results
(Aspartate Transferase (AST) Blood Test:
What It Is, Procedure & Results, n.d.)

Storage at 4°C, AST activity in serum is stable for up

to 48 hours. If samples are to be retained
for a longer period, they must be frozen
(Rifai et al., 2018, p.414).

Day-to-day 5-10% variation/day

Muscle injury Significant elevation

6. What are other methods of AST determination?

Colorimetric analysis The colorimetric analysis principle is based on the interaction of an

Cagayan State University - Andrews Campus, Tuguegarao City
CLINICAL CHEMISTRT 2 Laboratory | TTh, 5:00 PM - 8:00 PM
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Wilson Leal Laranang, RMT, MSc Not started

enzyme and a substrate to create and colorimetrically evaluate a
colored, light-absorbing complex by adding another reagent upon
stopping the enzyme process (Huang et al., 2006).
Since the 1950s, colorimetric analysis has gradually been replaced
by spectrophotometric analysis due to its limitations, including the
Spectrophotometric
need to make calibration curve and stop enzyme activity.
Meanwhile, there are apparent advantages to spectrophotometric
measurement
measurement, such as continuous testing and a large
measurement range (Huang et al., 2006).
Because of its excellent sensitivity, low background signal level,
Chemiluminescence and wide range of quantifiable concentrations ranging over five
decades of magnitude, chemiluminescence detection was chosen
for GOT and GPT detection (Huang et al., 2006).
it really is important to keep in mind that the chromatographic
separating of the reaction mixture was more difficult due to the
Chromatography presence of molecules with very similar chromatographic activity
but very different hydrophobicity and charge. As a result, optimizing
the chromatographic conditions for the detection of both enzymes
in a single blood sample is important (Huang et al., 2006).
Fluorimetric techniques have been established by Rietz & Guilbault
(1975) for determining the activity of the enzymes GOT, GPT, and -
Fluorescence and
HBD in solution and via a solid-surface approach using Dade
Reagent Tablets. The rate of appearance of NADH fluorescence at
UV absorbance
455 nm (ex=365 nm) was determined in each case and equated to
the concentrations of these enzymes in control serum and blood
serum.

References.
Cagayan State University - Andrews Campus, Tuguegarao City
CLINICAL CHEMISTRT 2 Laboratory | TTh, 5:00 PM - 8:00 PM
Instructor Status Notes

Wilson Leal Laranang, RMT, MSc Not started

Aspartate Transferase (AST) Blood Test: What It Is, Procedure & Results. (n.d.). Cleveland

Clinic.

https://my.clevelandclinic.org/health/diagnostics/22147-aspartate-transferase-ast

AST/ASAT/GOT. Colorimetric determination of serum GOT. (n.d.). Internet.

https://www.biotec-co.com/images/products/clinical_chemistry/kits/chemistry/AST

%20colorimetric.pdf

AST/Liquiform. (n.d.). Ref.:109. Internet,

https://labtest.com.br/wp-content/uploads/2016/09/

Ref_109_RevMaio2013_Ref180613_Eng.pdf

Cooper, A. (2019, March 19). Aspartate Aminotransferase. Welcome to the Exeter Clinical

Laboratory International website.

https://www.exeterlaboratory.com/test/aspartate-aminotransferase/

Huang, X. J., Choi, Y. K., Im, H. S., Yarimaga, O., Yoon, E., & Kim, H. S. (2006, July 31).

Aspartate Aminotransferase (AST/GOT) and Alanine Aminotransferase (ALT/GPT)

Detection Techniques. Sensors, 6(7), 756–782.

https://doi.org/10.3390/s6070756
Cagayan State University - Andrews Campus, Tuguegarao City
CLINICAL CHEMISTRT 2 Laboratory | TTh, 5:00 PM - 8:00 PM
Instructor Status Notes

Wilson Leal Laranang, RMT, MSc Not started

Irjala, K. M., & Grönroos, P. E. (1998, January). Preanalytical and analytical factors affecting

laboratory results. Annals of Medicine, 30(3), 267–272.

https://doi.org/10.3109/07853899809005854

McPherson, R.A. & Pincus, M.R. (2017). Henry’s Clinical Diagnosis and Management by

Laboratory Methods. 23rd Edition. Elsevier. Philadelphia, USA.

Policarpio, J.M. (2021). Checkpoint. Clinical Chemistry.

Rietz, B., & Guilbault, G. (1975, July). Fluorimetric assay of serum glutamate oxaloacetate

transaminase, glutamate pyruvate transaminase and α-hydroxybutyrate dehydrogenàse

by solution an. Analytica Chimica Acta, 77, 191–198.

https://doi.org/10.1016/s0003-2670(01)95170-0

Rifai, N. et al. (2018). Tietz Texbook of Clinical Chemistry and Molecular Diagnostics. Sixth

Edition. Elsevier. St. Louis, Missouri, USA.

SGOT Test Procedure. (2019, December). Lab Tests Guide.

https://www.labtestsguide.com/sgot-test-procedure

Yazdi, P. (2021, January 21). High/low aspartate aminotransferase levels + function & diseases.

SelfDecode Labs.

https://labs.selfdecode.com/blog/aspartate-aminotransferase-ast/