Purification and characterisation of phytochemicals extracted from Rhizophora mucronata : Their efficacy against Pseudomonas aeruginosa infection in Catla catla

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Abstract
The aim of this work is to purify and characterise bioactive molecules from various solvent extracts prepared with the leaves of Rhizophora mucronata , a mangrove plant. In vitro assessment of antibacterial activity of n -hexane, diethyl ether, ethanol, and aqueous extracts (AEs) against Bacillus subtilis (0.4 ± 0.1, 1.1 ± 0.3, 0.9 ± 0.1, and 1.2 ± 0.7 mm), Staphylococcus aureus (1.2 ± 0.1, 2.3 ± 0.5, 3.2 ± 1.2, and 4.4 ± 1.7 mm), Klebsiella pneumoniae (0.5 ± 0.3, 1.7 ± 0.4, 3.9 ± 1.4, and 3.9 ± 0.9 mm), and Pseudomonas aeruginosa (0.2 ± 0.1, 2.5 ± 0.6, 3.7 ± 0.5, and 4.2 ± 1.3 mm) has proven to act as potent similar to that of commercial antibiotic. Corresponding to antibacterial activity result, antioxidant activity was also higher for AE (75.97% ± 0.5%) compared to remaining extracts (73.77% ± 2.5%, 69.99% ± 0.4%, and 65.24% ± 1.8%) at a concentration of 500 μg·mL −1 . Considering both the activities, AE was selected for separation of bioactive molecule by deploying two purification techniques – column chromatography and preparative thin layer chromatography consecutively. After purification, the sample was injected into mass spectrometer and the presence of N - p -coumaroyltyramine and luteolin with excellent antibacterial and antioxidant properties (13.1 ± 2.2 mm and 80.9% ± 2.3%, respectively) were recognised. Further, in vivo investigations on Catla catla fingerlings infected with P. aeruginosa indicated that partially purified extracts cured the infection and resulted in a 100% survival rate.

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