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Development and validation of a new method for extraction and quantification of Oleuropein in olive leaves by HPLC-DAD
Letícia Barbieri Estrada
Departamento de Ciência de Alimentos / FEA / UNICAMP
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Create a topicOlive trees are plants native to the Mediterranean, known for their products, olive oil, and olives. However, several by-products are discarded, such as olive leaves, which have in their composition several phenolic compounds, predominantly oleuropein, a compound beneficial to health. This work aimed to develop a DLLME (liquid-liquid dispersive micro-extraction) method for the extraction and quantification of Oleuropein in olive leaves cultivated in Delfim Moreira - MG, during their pruning period. Three cultivars were evaluated (Arbequina, Grapollo, and Koroneiki) using two drying methods (oven at 35ºC and lyophilization). An HPLC-DAD was used to validate the method and quantify Oleuropein, injecting 1 to 10 µL of extract, depending on the analyte concentration. The validated method had a detection limit of 0.1 mg.kg-1, with recoveries ranging from 101.61 to 112.56% and precision from 1.68 to 12.96%. The curve with the Oleuropein standard presents good linearity, with regression equation y = 9.4772 x - 1.3252 and R2 = 0.9986. The results show that the oven-dried cv. Arbequina is below the limit of quantification, on the other hand, the lyophilized contains 2.02 ± 0.16 mg.kg-1. For cv. Grapollo, the results were 0.50 ± 0.03 mg.kg-1 and 33.87 ± 0.48 mg.kg-1 for oven and lyophilized samples, respectively. Finally, the oven-dried cv. Koroneiki has 1.00 ± 0.10 mg.kg-1 of oleuropein, compared to 3.58 ± 0.05 mg.kg-1 for lyophilized. Thus, it can be observed that drying the samples in oven favors the degradation of the compounds of interest and that the method is suitable for quantifying Oleuropein.
Amanda Mellissa B Oliveira
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