Agarwood-producing plants which only available in the East part of Indonesia is Gyrinops versteegii. Traditionally the plant has been used by the local people to cure many diseases including tumor, diabetes, malaria, etc. This study aimed to evaluate the antioxidant activity of the old and young leaves of G. versteegii and their cytotoxic activities on HeLa and Vero cell lines. The leaves of G. versteegii was collected from Bogor Botanical Gardens. Leaves samples were dried and ground, prior to extraction with various solvents i.e. chloroform, methanol and water. The antioxidant activity was determined using DPPH methods, while the cytotoxic activities against HeLa and Vero cell lines were done using MTT assay at a serial concentration of 10 µg · mL−1 to 2 000 µg · mL−1. The most potent extract was then fractionated using VLC with a mixture of solvents elution. The VLC result fractions were subjected again for antioxidant assay and cytotoxic assay. The phytochemical profiles of the potent fraction were determined using TLC. The results showed that the chloroform extract of the old leaves had the highest antioxidant activity with IC50 of 11.66 ± 2.11 μg · mL−1. On the other hand, the chloroform extract of young leaves has highest cytotoxic activity against HeLa cell line with IC50 of 22.71 ± 1.31 μg · mL−1. The antioxidant activities of all the VLC’s fractions showed a significant lower compared to that of the crude extract. This was indicating the possible of synergism effect from the bioactive compounds in the crude extract. The bioassay-guided fractionation of the chloroform extract of young leaves showed that the most active fraction was obtained from with a solvent mixture of chloroform:ethyl acetate (70:30) with IC50 of 11.37 ± 2.67 μg · mL−1. This fraction did not show cytotoxicity on Vero cell lines. TLC analysis showed that this fraction contained phenolic, flavonoids and terpene as the bioactive compounds.

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