GAS CHROMATOGRAPHY-MASS SPECTROMETRY ANALYSIS AND ANTIOXIDANT ACTIVITY OF PUNICA GRANATUM L. PEELS AND ITS ROLE AS IMMUNOSTIMULANT AGAINST SCHISTOSOMA MANSONI INFECTION IN BIOMPHALARIA ALEXANDRINA
DOI:
https://doi.org/10.22159/ajpcr.2017.v10i1.15107Abstract
Objective: To evaluate the antioxidant activity and chemical composition of Punica granatum L. and test it as immunostimulants against Schistosoma mansoni infection to Biomphalaria alexandrina snails.
Methods: Antioxidant activity was evaluated by measuring the free radical scavenging activity of the 90% defatted methanol extract (90% DM) of P. granatum peels and its sub-derived fractions was evaluated via 2,2'-diphenyl-1-picrylhydrazyl and its chemical constituents were identified via gas chromatography-mass spectrometry (GC-MS) analysis. B. alexandrina snails were exposed to pomegranate extracts (PEs) for 1 month before their challenging with S. mansoni miracidia. Infection rates, immunological and histological parameters were, then, evaluated in PE-exposed snails and compared to controls.
Results: The antioxidants activities of PE, expressed as scavenging concentration at 50%, were in the following order; 90% DM (12.45) ˃n-butanol(15.59) ˃ethyl acetate (21.36) ˃water (49.16) µg/ml, compared to 7.50 µg/ml for ascorbic acid. The infection rates of PE-exposed snails were 20%,50%, 60%, 70%, and 80%, respectively, for 90% DM, n-butanol, ethyl acetate, water, and dichloromethane extracts compared to 95% in control snails. The number of amoebocytes showed a significant increase, clear differentiation, and size increment in exposed snails compared to controls. Moreover, hermaphrodite glands histology shows a full maturity in the formation of reproductive cells in PE-exposed snails. The GC-MS analysis of the 90% DM extract revealed the presence of 36 compounds representing 93.1% of the total composition. Piperidin-4-ol, 1,3-dimethyl-2,4,6-triphenyl (19.87%), and 6,11-dihydroxy-5,12 naphthacenequinone-1-carboxylic acid (7.80%) were the major components.
Conclusion: The identified compounds in 90% DM extract of P. granatum may be responsible for the high antioxidant activity of the fruit and it may account for its immunostimulatory effect against S. mansoni infection in B. alexandrina.
Keywords: Punica granatum L., Antioxidant activity, Biomphalaria alexandrina, Schistosoma mansoni, Infection rate, Gas chromatography-mass spectrometry.
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