Martins Gabriel Rocha, Monteiro Alvaro Ferreira, do Amaral Felipe Rafael Lopes, da Silva Ayla Sant'Ana
Laboratório de Biocatálise, Instituto Nacional de Tecnologia, Ministério da Ciência, Tecnologia e Inovações, Rio de Janeiro, 20081-312 RJ Brazil.
Programa de Pós-graduação em Bioquímica, Departamento de Bioquímica, Universidade Federal do Rio de Janeiro, Rio de Janeiro, 21941-909 RJ Brazil.
J Food Sci Technol. 2021 Dec;58(12):4693-4702. doi: 10.1007/s13197-020-04959-5. Epub 2021 Jan 18.
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 (R = 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 NaCO 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.
广泛用于定量多酚的方法——福林-西奥尔特法(FC法)——不能不加区分地应用,因为不同基质可能会影响该测定的准确性。因此,本研究旨在对FC法进行调整,以用于阿萨伊种子提取物,这是一种富含单宁的提取物,在各种治疗中具有潜在应用价值。首先,建立了方法前标准化,以确定诸如读取波长(765nm)、反应时间(30分钟)和参考物质(连苯三酚)等参数。在验证步骤中,调整后的方法对分析物呈线性响应(R = 0.9910),确保了其选择性(线性和选择性曲线在统计学上平行)和准确性(99.18 - 101.43%)。此外,该方法在评估的两个水平(重复性和中间精密度)上被证明是精确的(RSD≤2.63%),并且在碳酸钠浓度和反应时间变化方面是稳健的(RSD≤4.45%)。还计算了检测限和定量限(分别为9.9μg/mL和33.1μg/mL)。还评估了基于络合剂对单宁进行选择性沉淀的报道来进行单宁定量的额外步骤;然而,观察到非特异性沉淀,降低了结果的准确性。我们的工作成功地调整并验证了一种用于阿萨伊种子提取物总酚定量的方法,结果为38g连苯三酚当量/100g提取物。