Pharmacological, charactreization and safety evaluation Rhizophora mucronata Lam. methanolic extract for aphrodisiac potential in rats

Pharmacological studies were conducted to evaluate the potential aphrodisiac activity of mangrove plant in coastal area of Sabah, Rhizophora mucronata methanol extract (RM methanol extract) or commonly called ‘Red Mangrove’ using animal model. The study was initiated based on observation of proboscis monkeys in Sabah active sexual activity and high sex ratio towards the female (sex ratio 1:8.4) and their diet in consuming local plant (mangrove) that believed to contribute to the high sexual activities. Young leaves of RM collected around coastal area of Sabah was authenticated at the Sabah Forestry Department, Sandakan, Sabah with a voucher specimen, Rhizophora mucronata (Bakau Kurap); SAN 149220. The leaves were oven dried at 40 ± 3 °C overnight, powdered and subjected to 50 % Soxhlet extraction using methanol: water. The extract was filtered, rotary evaporated and freeze dried at 4 °C to remove excess solvent. The prepared extract was characterized for contents of total flavonoid, steroid and saponin. It was also furtherer analysed for determination of active compound using HPLC, ELSD-HPLC dan LCMS/MS. The investigation of aphrodisiac potentials on RM leave extract were determined by assessing the sexual potency enhancement. Assessments were done on the sexual desire (mounting behaviour test), sexual motivational (Partner Preference test), sexual performance (Sexual Behaviour Test) and quantification of reproductive hormones (Luteinizing hormone (LH), Follicle stimulating hormone (FSH) and Testosterone(T)), in animals of four groups (n=6) namely, Control (RO), RM extract (300 mg/kg), E. Longifolia extract (500 mg/kg) and L-Dopa (100 mg/kg) given orally (p.o). Pharmacokinetics of catechins were determined using rats plama following single oral dose of RM extract in the rat. Safety evaluation was conducted on RM methanol extract with, Heavy Metal Test, Microbial Test, Bacterial Reverse Mutation Test, Cytotoxicity Test and Acute Oral Toxicity Test and Subacute Toxicity test in animals RM extract 1000 (mg/kg). Aphrodisiac studies of RM methanol extract (300 mg/kg) have shown that the extract significantly increased the serum testosterone level in sexually experience rats. Sexual desire (libido) and motivation was improved as evidenced by the reduction in mount latency. Whereas sexual performance is enhanced by the reduction of intromission latency. From HPLC analysis, fraction collected matches with the catechin standard. Further analysis using LCMS/MS analysis, it has confirmed that compounds identified are Catechin. The biologically active compounds identified through steroidogenesis study showed that catechin found in the extract increases testosterone level. As part of a safety evaluation of RM extract, no reverse mutagenic potential at the highest concentration of 5000 µg/plate with and without metabolic activation system under the present test conditions. Acute oral toxicity of RM methanol extract was classified as Category 5 according to the GHS for the classification of chemicals as the LD50 is greater than 2000 mg/kg body weight. In the 28-day repeated oral dose toxicity of RM methanol extract was evaluated at 1000 mg/kg per day dose levels. The results showed that no significant toxicological changes were observed when 1000 mg/kg per day of RM methanol extract was administered to rats. Body weights and food consumptions showed constant increment, meanwhile, heart and kidney weights were observed with slight hypertrophy of heart and tubular necrosis of kidney. Besides this observation, no other sign of toxicity was observed. Based on these results, it can be concluded that the no observed adverse effect level of RM methanol extract is 1000 mg/kg per day. Therefore, the use of RM is expected to be safe and pharmacologically effective in animals, thus, represent safety data for humans.

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