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RPM for 10 minutes. The upper organic layer was collected and reading was taken at 532nm using
butanol : pyridine (15:1) as blank.

2.7. Biochemical estimation
The blood sample collected was kept in an anticoagulant tube for 45 minutes at room temperature and
then was centrifuged at 7200 rpm at 4 C for 15 minutes and serum was separated. This serum was
0
then used for various biochemical estimations mainly Serum glutamate pyruvate transaminase activity
(SGPT) and Serum glutamate oxaloacetate transaminase (SGOT).

2.7.1. Serum glutamate pyruvate transaminase activity (SGPT)
Human serum contains several transaminases of which aspartate transaminase and alanine
transaminase are of clinical significance. An elevated level of serum transaminase is linked with the

hepatic injury and is widely used to study the extent of liver damage. They are used for the diagnosis
of severe hepatitis and liver damage due to various drugs and chemicals. ERBA SGPT kit was used

for the determination of transaminase level in the serum and the method used was one specified by the
International Federation of clinical chemistry (IFCC). Absorbance was measured in autoanalyzer.
Multiple readings for each sample were taken at a fixed interval of 1 minute.

The absorbance change per minute (ΔA/min) were converted into international units of activity using
following formula
3
IU/L = (ΔA/min) × T.V. × 10
S.V. × Absorptivity×P
Where: T.V. = Total reaction volume in µL

S.V. = Sample volume in µL
Absorptivity = Millimolar absorptivity of NADH at 340 nm = 6.22
P = Cuvette pathlength =1cm

Activity at 37º (IU/L) = (ΔA/min)× Factor(1768)
c
2.7.2. Serum glutamate oxaloacetate transaminase (SGOT).

SGOT is an enzyme located in the cytosol of the liver cell. In terms of abundance liver is the second
most abundant organ having SGOT. It’s clinical significance being that its level in the blood serum

increases in case of hepatic injury and hence its level is used to detect for the possible damage to the
liver (Reitman and Frankel, 1957). ERBA SGOT kit was used and standard procedure as laid out by
International Federation of clinical chemistry (IFCC) was strictly followed. The absorbance was

measured using autoanalyzer and multiple reading of each sample was taken at an interval of 1
minute. The formula stated above in the SGTP section was used to convert our readings into
international units of activity.

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