Isolation and Identification of Bioactive Fractions from Mangrove Plants Hibiscus tiliaceus (Bhola) and Derris trifoliata (Kalilata), targeting Diuretic and Laxative Drug Discovery.

This project work was designed to investigate different fractionated stem extracts (in ethanol, n-hexane, chloroform, ethyl acetate and water) of two mangrove plants Hibiscus tiliaceus (Bhola) and Derris trifoliata (Kalilata) for diuretic and laxative potentials and their phytochemical nature. In diuretic test, furosemide (5 mg/kg body weight) was used as standard drug and the parameters like volume of urine up to 6^th hour, pH, density, conductivity and electrolyte contents (Na⁺, K⁺, Cl^-) of collected urine were measured. Among the four fractions, ethyl acetate fraction of stem of Hibiscus tiliaceus, showed significant (p<0.001) diuretic activity at dose 200 mg/kg and 400 mg/kbw compared with control. For stem of Derris trifoliata, chloroform fraction demonstrated more diuretic activity at dose 75 mg/kg and 150 mg/kbw. Upon urine analysis for ions, pH and conductivity, both the samples were found to work as loop diuretics mechanism, having carbonic anhydrase inhibitory effect also. In laxative activity test, bisacodyl (10 mg/kg body weight) was used as standard drug and a significant percent increment of soft fecal output (p<0.001) was observed for chloroform fraction of Hibiscus tiliaceus. Then one of the most bioactive fractions for each plant (i.e., ethyl acetate fraction for Hibiscus tiliaceus and chloroform fraction for Derris trifoliata) was subjected to column chromatography. One single spot (R_f value: 0.4, in mobile phase 20% EtOAc:80% n-hexane) was isolated (for 1.25 mg for Hibiscus tiliaceus) and in case of Derris trifoliata the R_f value of isolated single spot was 0.5 (mobile phase dichloromethane 100%) and the quantity was 1.15 mg. Phytochemical analysis of fractionated extracts for both the plants exhibited the presence of steroids, terpenoids, flavonoids, glycosides, phenolics. Further spectroscopic techniques (NMR, Mass) may reveal the chemical structure of these pure fractions.

• Research Project