Optimization of the extraction of phytometabolites with antioxidant and anti-inflammatory properties from leaves of Casearia sylvestris

MATIAS DAMIAN DUQUE DE ARCE; TORRES ANA MARIA; JUAN JOSE RUIZ DIAZ ; RICCIARDI GABRIELA; OJEDA GONZALO

Mar del Plata

Congreso; LI reunión anual de la asociación Argentina de Farmacología Experimental; 2019

Casearia sylvestris Sw. is a tree widely distributed in South America. In Brazil´s popular medicine, the use of this plant is correlated with its pharmacological properties including anti-inflammatory, anti-ophidian and anti-ulcer activities. However, the development of a separation technique of bioactive compounds in the herbal extracts is particularly difficult, due to the high complexity of the matrix.The objective of this work was to determine the optimal conditions for the extraction of bioactive compounds with the software Design-Expert® 11, using a mixture design of three solvents (water/ethanol/acetone).Leaves of C. sylvestris were collected in Paso de la Patria (Corrientes, Argentina). The samples were dried, powdered and extracted by maceration under agitation for 2 days in the corresponding solvent. The extracts were filtered and concentrated under reduced pressure. Total phenolic, flavonoids and tannins contents were measured using Folin-Ciocalteau, aluminum trichloride and vanillin methods respectively. Antioxidant activity (AOA) was determined by DPPH and FRAP assays. In vitro anti-inflammatory activity (AIA) was investigated by the hypotonic hemolysis inhibition method.Ten different mixtures were evaluated by response surface methodology, in their ability to extract the bioactive compounds. To maximize biological activities a graphic and numerical optimization was performed, using desirability function (D). It was found that the maximum AOA was obtained by optimizing both, phenolic content and DPPH/FRAP assays (D=1), in extracts with acetone:water 1:1. Evaluation of hemolysis inhibition indicated that the highest AIA, was achieved with flavonoids (D=0.87) and tannins (D=0.96), obtained with ethanol.In conclusion, from the analysis of these results, we can predict that in order to optimize the extraction of active metabolites with AOA and AIA, the extracts should be obtained with acetone:water 1:1 mixture and ethanol respectively.