INCITAP UNLPam-CONICET, Instituto de Ciencias de la Tierra y Ambientales de La Pampa, Departamento de Química, FCEyN, Universidad Nacional de La Pampa (UNLPam), Av. Uruguay 151, (6300) Santa Rosa, La Pampa, Argentina.
INCITAP UNLPam-CONICET, Instituto de Ciencias de la Tierra y Ambientales de La Pampa, Departamento de Química, FCEyN, Universidad Nacional de La Pampa (UNLPam), Av. Uruguay 151, (6300) Santa Rosa, La Pampa, Argentina.
Food Chem. 2017 Aug 15;229:44-49. doi: 10.1016/j.foodchem.2017.02.029. Epub 2017 Feb 13.
The enzymatic deglycosylation of the plant flavonoid rutin (quercetin-3-O-(6-O-α-l-rhamnopyranosyl-β-d-glucopyranoside) is usually assessed by means of high performance liquid chromatography (HPLC). We have developed a spectrophotometric method for the quantification of the released quercetin. After the enzymatic reaction, quercetin is extracted with ethyl acetate, and subsequently oxidized under basic conditions. The absorbance of quercetin autooxidation products at 320nm was correlated with the quercetin concentration by linear regression (molar extinction coefficient 23.2 (±0.3)×10Mcm). With this method, rutin-deglycosylation activity in buckwheat flour and a commercial naringinase was measured, and showed no significant differences with the results obtained by HPLC. The convenience of this method resides on the enzymatic activity quantification using the natural substrate by UV-visible spectrometry. Moreover, the simplicity and speed of analysis allows its application for a large number of samples.
植物类黄酮芦丁（槲皮素-3-O-（6-O-α-L-鼠李吡喃糖基-β-D-吡喃葡萄糖苷）的酶解糖化通常通过高效液相色谱法（HPLC）进行评估。我们开发了一种用于定量释放的槲皮素的分光光度法。在酶反应后，用乙酸乙酯提取槲皮素，然后在碱性条件下氧化。在 320nm 处，槲皮素自动氧化产物的吸光度与槲皮素浓度通过线性回归相关（摩尔消光系数 23.2（±0.3）×10Mcm）。使用该方法，测定了荞麦粉和一种商业柚皮苷酶中的芦丁去糖化活性，其结果与 HPLC 测定的结果无显著差异。该方法的便利性在于使用 UV-可见分光光度法通过天然底物进行酶活性定量。此外，分析的简单性和速度使其适用于大量样品。
