Materials and Methods: Crude methanolic fraction of E. suberosa (Roxb) bark and its respective fractions were screened for the presence of different phytochemicals with different reagents. On the basis of increasing order of polarity, different organic solvents were used to obtain different fractions. Enzymatic studies were performed on crude methanolic extract of the plant. All the assays were performed under standard in vitro conditions.

Results: The phytochemical analysis shows the presence of alkaloids, phenols, triterpenoids, phytosterols, and flavonoids. Phenolic compounds and flavonoids are the major constituents of the plant. In anticholinesterase assay, the percent inhibition of standard drug (eserine) was 91.27 ± 1.17 and the half maximal inhibitory concentration (IC50) was 0.04 ± 0.0001. For α-glucosidase inhibition, the IC50 value for Dichloromethane fraction was 8.45 ± 0.13, for Methanol fraction it was 64.24 ± 0.15, and for aqueous fraction it was 42.62 ± 0.17 as compared with standard IC50 that is 37.42 (acarbose). Furthermore, results show that all fractions have potential against anti-urease enzyme, but DCM fraction of crude aqueous extract has significant IC50 value (45.26 ± 0.13) than other fractions.

Materials and Methods: The bark was extracted using different solvents, for example, dichloromethane, ethyl acetate, methanol, and aqueous for obtaining the organic fractions. These organic fractions were then evaluated for their cytotoxic and antimicrobial activity compared with the standard. Cefixime was used as the standard for antibacterial assay, whereas clotrimazole was used as the standard for antifungal activities. Bacterial strains used were Staphylococcus aureus and methicillin-resistant S. aureus (MRSA), whereas for antifungal activities Candida albicans, Candida parapsilosis, and Candida krusei strains were used.

Results: The organic fractions obtained were evaluated for their cytotoxic and antimicrobial activities. In cytotoxic assay (Brine shrimp lethality assay), dichloromethane fraction was the most potent with LD50 of 47.63, whereas aqueous, methanol, and ethyl acetate fractions showed LD50 of 121.74, 422.2, and 201.96, respectively. Similarly, for antibacterial assay, dichloromethane fraction showed 32.2mm zone of inhibition against MRSA in comparison with standard cefixime (zone of inhibition, 30.5mm). A minimal zone of inhibition with crude saponins (13.1 and 12.2mm) was observed against C. albicans in comparison to standard (cefixime) with a zone of inhibition of 28.5mm. No prominent results were observed against C. parapsilosis and C. krusei strains.

Methods: Chemical profiling of P. blanda was carried out using gas chromatography mass spectrometry (GCMS) followed by isolation of bioactive compounds by column chromatography. DPPH• and FRAP assays were used to evaluate antioxidant activity and the MTT assay was performed to estimate the cytotoxicity activity against three cancer cell lines, namely MCF-7, HL-60 and WEHI-3, and three normal cell lines, MCF10A, WRL-68 and HDFa.

Results: X-ray crystallographic data for peperomin A is reported for the first time here and N,N'-diphenethyloxamide was isolated for the first time from Peperomia blanda. Methanol and dichloromethane extracts showed high radical scavenging activity with an IC50 of 36.81 ± 0.09 µg/mL, followed by the dichloromethane extract at 61.78 ± 0.02 µg/mL, whereas the weak ferric reducing activity of P. blanda extracts ranging from 162.2 ± 0.80 to 381.5 ± 1.31 µg/mL were recorded. In addition, petroleum ether crude extract exhibited the highest cytotoxic activity against all the tested cancer cell lines with IC50 values of 9.54 ± 0.30, 4.30 ± 0.90 and 5.39 ± 0.34 µg/mL, respectively. Peperomin A and the isolated mixture of phytosterol (stigmasterol and β-sitosterol) exhibited cytotoxic activity against MCF-7 and WE-HI cell lines with an IC50 of (5.58 ± 0.47, 4.62 ± 0.03 µg/mL) and (8.94 ± 0.05, 9.84 ± 0.61 µg/mL), respectively, compared to a standard drug, taxol, that has IC50 values of 3.56 ± 0.34 and 1.90 ± 0.9 µg/mL, respectively.