Flavonoids from Passiflora cincinnata (Passifloraceae): HPLC-DAD-MSn characterization, validation of an HPLC-DAD method for quantification and CPC isolation

Passiflora cincinnata Mast. is a native tree from the Brazilian Caatinga biome, popularly known as “maracujá do mato” and “maracujá da Caatinga”. This species has been used in folk medicine as a sedative and antihypertensive. This study presents the chemical characterization of P. cincinnata using a HPLC-DAD-MSⁿ approach. Due to their nutraceutical and pharmaceutical interest, we also describe the validation of a simple and efficient HPLC-DAD method for the quantification of flavonoids. The ethanolic extract was submitted to centrifugal partition chromatography (CPC) for the isolation of flavonoids. HPLC-DAD-MSⁿ analysis allowed the annotation of nine chemical constituents with chemotaxonomic importance, including orientin, isoorientin, vitexin and isovitexin, which are considered as analytical markers of Passifloraceae species. Thereafter, we developed and validated an HPLC-DAD method for the quantification of the flavonoids orientin, vitexin and isovitexin in order to contribute to the quality control assessment of P. cincinnata samples. The analytical method has proven to be linear, accurate, robust, reproducible and sensitive to all investigated flavonoids. Two flavonoids (isoorientin and isovitexin) were isolated in a single and fast CPC run followed by preparative HPLC, and their structures were confirmed by NMR analysis. This is the first report of a validated method for flavonoid quantification in this species, as well as the isolation of flavonoids using CPC; it may also be useful for nutraceutical and pharmaceutical industries in the development of new Passiflora products.