EVALUATION OF THE GENOTOXICITY OF EURYCOMA LONGIFOLIA AQUEOUS EXTRACT (PHYSTA®) USING IN VITRO AMES TEST AND IN VIVO MAMMALIAN MICRONUCLEUS TEST
Keywords:
Eurycoma longifolia, Ames test, Micronucleus assay, GenotoxicityAbstract
Objective: In this study the genotoxic potential of Eurycoma longifolia aqueous extract (PHYSTA®) was investigated by the in vitro Ames test (Salmonella/microsome mutation assay) and the in vivo mouse peripheral blood cell micronucleus test. Eurycoma longifolia also known as Tongkat Ali in Malaysia, is traditionally used in South East Asia to treat fever, intestinal worms, mouth ulcers, headache, erectile dysfunction and many other general pains. It is traditionally used as a health tonic and anti-stress remedy. Recent studies of Eurycoma longifolia pharmacological profile have revealed antioxidant properties and other potentially useful biological activities thereby lending some scientific support to its use in folk medicine. Evaluation of the genotoxic potential is one of the most important nonclinical safety studies required for registration and approval for marketing of pharmaceutical products.
Methods: The Salmonella/microsome mutation assay (TA 98, TA 100, TA 102, TA 1535 and TA 1537; plate incorporation method) was performed in the presence or in the absence of extrinsic metabolic activation (S9 mixture). In the mouse micronucleus assay, Eurycoma longifolia aqueous extract was administered intraperitoneally (100, 250 and 500 mg/kg body weight; single dose) to male and female NMRI mice (N= 5 per dose per sex) and peripheral blood was analyzed after 44 and 68 h after treatment using a flow cytometer to determine micro nucleated polychromatic (immature) erythrocytes.
Results: The results were expressed as the relative proportion of polychromatic erythrocytes among total erythrocytes (relative PCE). Tested at doses up to 5 mg/plate, the Eurycoma longifolia aqueous extract (PHYSTA®) was not toxic to Salmonella tester strains and did not increase the number of revertant colonies over the background incidence. In the mouse peripheral blood cell micronucleus assay, the extract did not alter the relative PCE, nor did it increase the incidence of micro nucleated polychromatic erythrocytes.
Conclusion: Based on the aforementioned findings, it is concluded that the Eurycoma longifolia aqueous extract (PHYSTA®) has no mutagenic potential and considered to be non-genotoxic with respect to clastogenicity.
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