A validated Folin-Ciocalteu method for total phenolics quantification of condensed tannin-rich açaí (Euterpe oleracea Mart.) seeds extract

The widely used methodology to quantify polyphenols-the Folin-Ciocalteu (FC) method-cannot be applied indiscriminately since different matrices may impair the assay's accuracy. Thus, this study aimed to adapt the FC method for the açaí seed extract, a tannin-rich extract with potential applications for various therapies. Firstly, a pre-method standardization was established to determine parameters such as reading wavelength (765 nm), reaction time (30 min), and the reference substance (pyrogallol). In the validation step, the adapted method responded linearly to the analyte (R2 = 0.9910), ensuring its selectivity (linearity and selectivity curves statistically parallel) and accuracy (99.18-101.43%). Furthermore, the method proved to be precise (RSD ≤ 2.63%) at the two levels assessed (repeatability and intermediate precision) and robust (RSD ≤ 4.45%) concerning variation on the Na2CO3 concentration and the reaction time. The limits of detection and quantification were also calculated (9.9 µg/mL and 33.1 µg/mL, respectively). An additional step for tannins quantification based on its reported selective precipitation by complexing agents was also evaluated; however, unspecific precipitation was observed, reducing the results' accuracy. Our work successfully adapted and validated a method for total phenolics quantification of açaí seed extract, resulting in 38 g of pyrogallol equivalent/100 g of extract.