Untargeted Bioassay Strategy for Medicinal Plants: In Vitro Antidiabetic Activity and 13C NMR Profiling of Extracts from Vitex negundo L
Bioassay-guided fractionation is the principal method for the identification of active constituents in medicinal plants. By design, this method aims to identify the most active compound in a complex mixture with the objective of discovering novel drug candidates. Described here is a complementary method for the identification of known bioactive compounds in medicinal plants which is untargeted and which takes advantage of the large NMR database of known natural products and availability of statistical software. This untargeted bioassay strategy is demonstrated as a proof of principle in the determination of the antidiabetic compounds in Vitex negundo L. Crude methanol and ethanol extracts, and chloroform, ethyl acetate and aqueous fractions of V. negundo L. were prepared and tested for their in vitro antidiabetic potential using the glucose diffusion retardation assay and the in vitro starch-amylase inhibition assay. The same crude extracts and fractions were profiled using 13C nuclear magnetic resonance (NMR) spectroscopy. The 13C NMR spectra of twelve known compounds from the semi-polar fraction of V. negundo – two iridoids, seven iridoid glucosides, two flavonoids and one flavonoid C-glucoside – were matched from the 13C NMR spectra of the extracts and fractions. The 13C NMR match factor values of the twelve compounds were used in the multivariate correlation analysis with antidiabetic activity using the glucose diffusion retardation activity and the starch-amylase inhibition assay. This method was able to correlate the seven iridoid glucosides with the antidiabetic activity, a result that would have been difficult to obtain using bioassay-guided fractionation.
Zabala MJB, Lagurin LG, Dayrit FM (2018) Untargeted Bioassay Strategy for Medicinal Plants: In Vitro Antidiabetic Activity and 13C NMR Profiling of Extracts from Vitex negundo L. Med Aromat Plants (Los Angeles) 7: 313. doi:10.4172/21670412.1000313