METHODOLOGY
The methodology adopted to assess the " Antioxidant levels
and hepatoprotective efficacy of Ashshifa-1000" is given below:
Qualitative analysis of Ashshifa-1000
Qualitative analysis of Ashifa-1000 was carried out for the presence
of protein, carbohydrate, tannins, alkaloids, steroids, saponins
and flavonoids.
Quantitative analysis of Ashshifa-1000
Ashshifa-1000 was quantitatively assessed for the presence of various
chemical components like protein, carbohydrate, tannins, flavonoids
and the minerals calcium, phosphorus and iron.
Estimation of antioxidants
Ashshifa-1000 was quantitatively assessed for the presence of various
enzymic antioxidants like catalase, peroxidase, superoxide dismutase
and polyphenol oxidase.
Non-enzymic antioxidants like ascorbic acid, riboflavin and tocopherol
were also analysed.
Determination of extent of inhibition of in vitro lipid
peroxidation
Extract of Ashshifa-1000 was prepared and the extent of inhibition
of lipid peroxidation was done by TBARS assay.
Ames Salmonella microsome assay
The mutagenic/antimutagenic effect of Ashshifa-1000 was tested using
the mutant bacterial strains of Salmonella typhimurium TA 98, TA
100 and TA 102 obtained from Prof B.N. Ames, Biochemistry Department,
University of California, Berkeley, California, U.S.A.
Assessment of the hepatoprotective efficacy of Ashshifa-1000
Preparation of the Ashshifa-1000 extract for administration
5.0g of the Ashshifa-1000 powder were taken and the extract was
prepared with 100ml alcohol. Evaporated off the alcohol completely.
To this added 50ml of 10% DMSO. 0.5ml of this (equivalent to 50mg
Ashshifa-1000) per kg body weight of mice was administered orally.
introduction of hepatotoxicity in experimental animals using
CCL4
Carbon tetra chloride (100%) was diluted with olive oil in the ratio
of 1:1 and injected at the
dose of 2ml/kg body weight by intraperitonial method. This amount
was given to the mice
groups III, V, VI and VII.
Grouping the experimental animals and the protocol
Fifty Swiss albino mice (7-8 weeks old) were obtained from Kerala
Agricultural University, Thrissur. They were divided into 7 groups
of 6 each as follows:
Group I: Control - m Normal healthy mice
Group II: Mice treated with olive oil only
Group III: Mice treated with GCL4 in olive oil (1:1)
Group IV: Mice treated with Ashshifa-1000 for 10 days
Group V: Mice treated with CC L4 followed by Ashshifa-1000 for 10
days.
Group VI: Mice treated simultaneously with CCL4 and Ashshifa- 1000
for one day only.
Group VII: Mice treated with Ashshifa-1000 for 4 days. Then CCL4
in olive oil was injected on the 5th day and continued the administration
of Ashshifa-1000 for 6 more days;
Assessment of liver enzymes in the experimental animals
The liver homogenate of the mice were prepared and the enzymes namely
aspartate transaminase, alanine transaminase, alkaline phosphatase
and lactate dehydrogenase were estimated. Liver glycogen, blood
glucose and serum bilirubin were also estimated.
Histopathological study
The animals were sacrificed by cervical dislocation and an autopsy
was carried out to obtain the liver of the animals. A portion of
the liver from mice of each group was fixed in 10% formaldehyde
and the histopathology was carried out,
RESULTS AND DISCUSSION
The observations made on the nutritional, biochemical and medicinal
properties of Ashshifa-1000 and the results obtained are presented
and discussed below:
Identification of the phytochemicals present in Ashshifa-1000
A systematic qualitative analysis of the aqueous and alcoholic extracts
of
Ashshifa-1000 was carried out to identify the phytochemicals present
in it. The result are given in Table I. |
