Anthocyanins and phenolic compounds in five ripening stages of Byrsonima ligustrifolia after extraction optimization

Byrsonima ligustrifolia is an astringent fleshy berry fruit whose colour changes during the maturation process from green to purple-black at over-ripe. Like other berries, it can be a rich source of antioxidant phenolics. In the present study, the best combination between three extraction solvents (acetone, methanol, and water) was determined, using simplex-centroid design with axial points and two replicates at the center point. After that, phenolic compounds and anthocyanins from B. ligustrifolia fruit in five ripening stages were studied by colorimetric assays, high performance liquid chromatography (HPLC) coupled to mass spectrometry (MS/MS), and HPLC equipped with diode array detector (DAD). Our results showed that acetone : methanol (20 : 80) worked better in extracting both compounds and, from the first to the last ripening stage, the content of total phenolic compounds decreased to 57% and total monomeric anthocyanins became 50 times more concentrated. HPLC-MS/MS allowed to identify cyanidin-3-glucoside, pelargonidin-3-glucoside, peonidin-3-glucoside, delphinidin-3-glucoside, gallic acid, protocatechuic acid, p-hydroxybenzoic acid, caffeic acid, p-coumaric acid, quercetin and catechin. Among these compounds, six were quantified by HPLC-DAD: cyanidin-3-glucoside, pelargonidin-3-glucoside, gallic acid, p-hydroxybenzoic acid, p-coumaric acid, and catechin.