Phytochemical Analyses and Biological Activities of Different Parts of Three Varieties of Labisia Pumila Benth

Labisia pumila (Myrsinaceae), also known as "Kacip Fatimah" (KF), has been used by many generations to induce and facilitate childbirth. There are three varieties of KF in Malaysia, namely, L. pumila var. alata, L. pumila var. pumila and L. pumila var.lanceolata. Recently, researches have demonstrated its estrogenic activity and high concentration of phenols. Due to ethnopharmacological application and lack of scientific information about bioactive compounds and possible biological activities of Labisia pumila Benth. with the potential to be applied in pharmaceutical, cosmetic and food industries, this study was conducted to evaluate the leaf, stem and root parts of three varieties of L. pumila for the presence of flavonoids, phenolics and antioxidant activities in extracts obtained by three solvents (methanol, ethyl acetate and water) with different polarities. The antimicrobial properties of extracts were assessed in reflux and microwave obtained extracts. Furthermore, qualitative and quantitative compositions of flavonoids and phenolics in all the extracts were analyzed using High-performance liquid chromotography (HPLC) system. Apart from that the anti-inflammatory and anticancer activities were determined in microwave methanolic extract. Finally, different levels of glasshouse irradiance were tested in order to examine possible changes in the total phenolic and flavonoid contents and antioxidant activities of all varieties of L. pumila. Similar results were obtained from two different methods (reflux and microwave) of extraction on biological activities but the microwave result was more effective to increase the yield in a shorter period while at the same time, using less solvent compared to the reflux extraction. Meanwhile, in all the three varieties of L. pumila Benth. the methanolic extract showed higher contents of phenolics and flavonoids as compared to ethyl acetate and boiling water extract. Results on the phenolics and flavonoids contents in the leaves, stems and roots of the three varieties showed significant difference (p<0.05). In all the varieties, the leaves contained higher phenolics and flavonoids as well as higher antioxidant activity compared to the roots and stems. Antioxidant activities of all varieties also showed significant difference (p<0.05) by DPPH and FRAP analyses. L. pumila var. alata contained higher antioxidants compared to var. pumila followed by var. lanceolata. Meanwhile, methanolic extract from the leaf of L. pumila var. pumila had higher total flavonoids content (1.53 and 2.77 mg rutin equivalent /g dry weight (DW) than L. pumila var. alata (1.32 and 2.49 mg rutin equivalent /g DW) and var. lanceolata (1.28 and 2.29 mg rutin equivalent /g DW) while the leaf of L. pumila var. alata recorded higher total phenolics content (2.65 and 3.48 mg gallic acid equivalent (GAE)/g DW)than L. pumila var. pumila (2.56 and 3.37 mg GAE/g DW) and var. lanceolata (2.43 and 3.23 mg GAE/g DW) using both the reflux and microwave extractions methods, respectively. High performance liquid chromatography (HPLC) analyses of phenolics and flavonoids in all three varieties revealed the presence of gallic acid, caffeic acid, rutin,quercetin, kaempferol and myricetin in all plant parts. Crude methanolic extract of all varieties had exhibited antibacterial activities against both Gram positive (Micrococcus luteus, Bacillus subtilis B145, Bacillus cereus B43, Staphylococcus aureus S1431) and Gram negative (Enterobacter aerogenes, Klebsiella pneumonia K36, Escherichia coli E256, Pseudomonas aeruginosa PI96) pathogens at low concentrations but with lower activity than kanamycin. The antifungal activity of methanolic extracts of all varieties against Fusarium sp., Candida sp. and Mucor sp. using the agar diffusion disc was moderate compared to streptomycin. The anti-inflammatory activity was determined using microwave extract of the leaf and root by in vitro nitric oxide (NO) inhibitory activities in a murin macrophage cell line (RAW 264.7). The results demonstrated strong inhibitory activity in NO inhibition by all varieties. Cell exposed to the extracts showed viability with a range of 50 to 90%. In vitro anticancer activities of extracts against two human cancer cell lines (MCF-7; MDA-MB-231) and Chang cell as a normal human hepatocyte could be categorized as moderate to weak by leaf and root methanolic extracts. The compounds present in the extracts were non-toxic, which render them as suitable potential therapeutics to be developed into anticancer drug. Exposure to different levels of light intensity had affected the phenolics, flavonoids and antioxidant activity of L. pumila where the highest concentration and activity were demonstrated under high irradiance of 630 compared to 310 μmol m−2 s−1, suggesting that the efficacy of L. pumila can be altered and enhanced under increasing light intensity. The highest amount of these components accumulated mainly in the leaves followed by the roots and then the stems. In conclusion, three varieties of L. pumila Benth. are a potential source of bioactive compounds endowed with useful biological activities such as antioxidant, antimicrobial,anti-inflammatory and anticancer properties.

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