Abstract
This experiment was conducted in order to purify some active ingredients of 70% alcoholic extract of aniseed by adsorption chromatography on silica gel and gel filtration on sepharose CL-6B column, using phosphate buffer (0.1M, pH 7.0) as eluent, as well as gel filtration on sephadex LH-20 where chloroform : methanol (3:1) was used as an eluent. Thin layer chromatography on silica gel was used as a criterion for the identification of purified fractions. Ultra violet transilluminator as well as vanillin-H2SO4 reagent were used to identify and develop of separated fractions, respectively. However, the fractions which purified on sephadex LH-20, showed an Rf value (0.71) which corresponded to the Rf values of anethole. To assess the estrogenic activity of purified fractions a dose of 150 mg/kg of body weight of collected fractions were given to immature female mice at three weeks age and administered orally for four weeks period. Twenty immature female mice were divided randomly into five groups. First group received crude extract of aniseed, while the second, third, and fourth groups received fractions purified on silica gel, sepharose CL-6B and sephadex LH-20 columns, respectively. Group five served as control, such treatments were continued until maturity, and then the animals were sacrified for studying weight changes in ovaries and uterus. The results of this study revealed that there is a significant increment (P<0.01) in the weights of ovaries in the treated groups as compared to the control, while fractions obtained from sepharose CL-6B had no significant effect (P>0.01). Also there is a significant increment (P<0.01) in the weights of uterus in the treated group with respect to fractions obtained from sephadex LH- 20 as compared to other treated groups and control.
Highlights
Herbs have been used for nutritive and medicinal purposes for centuries
The least significant difference (LSD) test was used to compare significance between the means [18]. The result of this experiment showed that chromatography of 70% alcoholic extract of anis seed on sephadex LH-20 column yield to some extent a homogenous symmetrical peak Figure (3)
Thin layer chromatography technique revealed that the bioactive fractions obtained from sephadex LH-20 column appears as rose color spot seen by using UV transilluminator and vanillin-H2SO4 reagent, which has Rf value equal to 0.71 .This Rf value identical to Rf value of anethole reported by [14]
Summary
Herbs have been used for nutritive and medicinal purposes for centuries. The World Health Organization estimated that 80% of the earth s inhabitants rely on traditional medicine for their primary health care needs, and most of this therapy involves the use of plant extracts or their active components [1]. [13] concluded that the administration doses (43 mg, 86mg /kg B.W.) of anise oil to male mice resulted in amarked improvement in reproductive capacity, which explained by the effect of treatment on spermatogenesis, probably by hormonal regulation across hypothalamic-pituitary-testicular axis. This experiment was aimed and designed to evaluate the estrogenic performance of the active ingredients of aniseed in albino immature female mice, by using chromatographic purification of bioactive componentes of alcoholic extract of aniseed on silica gel, sepharose CL-6B and sephadexLH-20, and identification of purified fractions
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