ANTISICKLING EFFECT AND ACTIVITY GUIDED FRACTIONATION OF DICHROSTACHYS CINEREA (Wight & Arn) ROOT AND STERCULIA SETIGERA (Del) LEAF METHANOLIC EXTRACTS ON HUMAN SICKLE RED BLOOD CELLS IN VITRO
ANTISICKLING EFFECT AND ACTIVITY GUIDED FRACTIONATION OF DICHROSTACHYS CINEREA (Wight & Arn) ROOT AND STERCULIA SETIGERA (Del) LEAF METHANOLIC EXTRACTS ON HUMAN SICKLE RED BLOOD CELLS IN VITRO
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Date
2016-02
Authors
ABDULLAHI, Baraka
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Abstract
The in vitro antisickling effects of the crude methanolic extract, ethyl acetate, n-hexane, butanol, and aqueous fractions of methanolic extracts of Dichrostachys cinerea and Sterculia setigera were determined using different concentrates on sodium metabisulphite-induced sickled red blood cells. The result showed that antsickling activity was both concentration and time dependent, with highest activity observed at the longest time interval of 120 min, with the exception of n-hexane concentrate of 0.1mg/ml and aqueous fraction of 0.2mg/ml of Dichrostachys cinerea having a percentage unsickling effect of 38.46±1.4% and 33.39±1.4%, respectively. The ethylacetate butanol, n-hexane, and aqueous concentrates of Dicrostachys cinerea fractions showed a significant antisickling difference (p<0.05) when compared with p-hydroxybenzoic acid, (PABA) as positive control. However, the butanol concentrate of Sterculia setigera did not show any significant difference when compared with the positive control. Osmotic fragility test was carried out in saline concentrations of 0.1, 0.2, 0.3, 0.5, 0.6, 0.9ml using extract concentrations of 0.5, 1.0, 2.0, and 2.5mg/ml. A significant difference was observed in percentage inhibition of lysis in ethylacetate fraction of Sterculia setigera for all saline concentration when compared with a standard drug, Ibuprofen. For butanolic fraction of Sterculia setigera, there is no significant difference in percentage inhibition of haemolysis when compared with Ibuprofen in all tested saline concentrations. N-hexane fraction of Sterculia setigera showed a significant difference in percentage inhibition of haemolysis, when compared with Ibuprofen. Also, a significant difference was observed in percentage inhibition of lysis when compared with Ibuprofen. For ethylacetate fraction of Dicrostachys cinerea, a significant difference was observed in all tested saline concentrations, when compared with Ibuprofen.
However, in the butanolic fraction, no significant difference in percentage inhibition of haemolysis was observed when compared to Ibuprofen. The n-hexane and aqueous fractions of Dicrostachys cinerea showed a significant difference in percentage lysis in all tested saline concentrations when compared with ibuprofen. The percentage methaemoglobin concentration was analysed using extract concentrations of 0.2, 0.4, 0.6, 0.8, and 1mg/ml. results showed that no significant difference was observed in all fractions of both test plants, when compared with the control. Elemental analysis was carried out using an AAS, the highest concentrations of Fe, Zn, Cu, and Cr were observed in aqueous fraction of Dichrostachys cinerea with a concentration of 287.20±0.00, 72.60±0.00, 57.20±0.00, 20.80±0.00 respectively, while the ethylacetate fraction has the least concentrations of the elements. Methanolic extracts of D. cinerea and S. setigera showed the highest antisickling activity. Aqeous fraction of S. setigera showed the highest methaemoglobin reduction property, while methanolic extract showed the best methaemoglobin reduction effect. Membrane protection activity was best in butanol fraction of D. cinerea, but methanolic extract of S. setigera had the highest membrane protection activity. The aqueous fraction of D. cinerea had the highest mineral concentration, while butanolic fraction of S. setigera had highest concentration of minerals. .
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A THESIS SUBMITTED TO THE SCHOOL OF POSTGRADUATE STUDIES, AHMADU BELLO UNIVERSITY, ZARIA IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE AWARD OF MASTER OF SCIENCE DEGREE IN BIOCHEMISTRY DEPARTMENT OF BIOCHEMISTRY, FACULTY OF SCIENCE, AHMADU BELLO UNIVERSITY, ZARIA, NIGERIA
Keywords
ANTISICKLING EFFECT,, ACTIVITY GUIDED FRACTIONATION,, DICHROSTACHYS CINEREA,, ROOT,, STERCULIA SETIGERA,, LEAF METHANOLIC EXTRACTS,, HUMAN SICKLE RED BLOOD CELLS,, VITRO