The Effect of Methanolic Extract and Ethyl Acetate Fraction of Kepel Fruit (Stelechocarpus burahol) to α-Glutathione S-Transferase Enzyme Concentration of Rat Liver and Blood Induced by CCl4

Abstract

Degenerative diseases are mostly caused by high free radical concentrations in the body. Kepel fruit
(Stelechocarpus burahol) is known to contain flavonoid compounds, a class of compounds that has free radical
scavenging activity in the body and could affect the activity of antioxidant enzymes. There have been reports
that methanolic extract and ethyl acetate fractions of Kepel fruit have the lowest IC50 values, compared to
extracts or fractions which are made using other organic solvents, on in vitro assays so that flavonoids are
suspected as responsible compounds to the free radical scavenging activity. This study aimed to identify the
presence of flavonoid content in methanolic extract and the fraction of ethyl acetate of Kepel fruit and also its
effect on rat alpha Glutathione S-Transferase (α-GST) enzyme concentration for the function to prevent hepatic
cell damage due to carbon tetrachloride exposure. The identification of flavonoids content used a series of
color chemical reaction tests and thin layer chromatography that used silica gel 60F254 as the solid phase and
mixture of ethyl acetate, formic acid, glacial acetic acid, and water (100: 11 : 11: 20 v/v) as the mobile phase.
This study used 30 Sprague-Dawley female rats (age 2-3 months) that were grouped into 5 groups as solvent
control (CMC-Na) group, negative control (CCl4) group, positive control (vitamin C) group, methanolic extract
group, and ethyl acetate fraction group. All groups were induced with carbon tetrachloride (CCl4) compounds
except for the CMC-Na solvent control group. Blood sampling at 0 h, 24 h, and 72 h, while liver sampling at 24
h and 72 h, with 3 samples each group. The analysis of rat α-GST enzyme concentrations used the
Enzyme-linked Immunosorbent Assay (ELISA) method. Based on the phytochemical test, the methanolic
extract and the ethyl acetate fraction of Kepel fruit have been identified contains flavonoids compound. Based
on the in vivo study, the positive control group's α-GST enzyme concentration was decreased, while the
negative control group's α-GST enzyme concentration was increased. Administration of methanolic extract
and ethyl acetate fraction also decreased both liver and blood rat α-GST enzyme concentrations, although
without significant correlation, and still could prevent the hepatic cell damage due to carbon tetrachloride
exposure.