Development of analytical method for quantification of chemical marker in ethanol extract from leaves of Campomanesia lineatifolia

The Campomanesia lineatifolia Ruiz & Pav. is native species to the Amazon west region and is a tree that can reach from 8 to 10m of height. It is popularly known as gabiroba and its fruits are edible and much appreciated3. In a previous study from our research group, we observed the presence of flavonoids in an ethanol extract from leaves of C. lineatifolia obtained by percolation. This extract was partitioned sequentially with hexane, dichloromethane, ethyl acetate and n-butanol, yielding a flavonoidic enriched fraction in the ethyl acetate step. In addition, we have observed a high antioxidant activity, suggesting a correlation with its ethnopharmacological indication as antidiarrheal, anti-gastric ulcer and healing1,2. Thus, the objective of this work was to develop an analytical method for quantification of the main constituents of the extract from leaves of C. lineatifolia. To develop the analytical method, we used an optimized ethanolic extract, flavonoids enriched, obtained from sheets of C. lineatifolia after extraction using reflux (EtOH R) and we evaluated parameters as preparation of the sample; organic modifier; oven temperature; flow; slope; time of analysis and separation, asymmetry and resolution of the peaks. An ultra efficiency UPLC Acquity System ® coupled to the detector UV / DAD and a standard Acquity UPLC ® BEH C18 column (2.1 x 100 mm d.i., 1.7 uM) with VanGuard ™ C18 pre-column (2.1 x 5 mm, 1.7 uM ), were used. The most efficient experimental condition found in a linear gradient system using a mixture of solvents water:acetonitrile (85:15; v / v) is described in Table 1. The obtained chromatogram using three chemical markers is observed in Figure 1.
The developed chromatographic conditions showed the selected chemical markers well separated in a short run time, symmetric and well resolved peaks. This methodology will contribute to the chemical and biological standardization of the extract from C. lineatifolia leaves, as well as the identification of the chemical markers for this species.