Screening and quantification of the enzymatic deglycosylation of the plant flavonoid rutin by UV-visible spectrometry

- Quantification of rutin-hydrolyzing enzymes using the natural substrate.
- Determination of rutin-deglycosylation activity in buckwheat flours and naringinase.
- Spectrophotometric method and HPLC method did not shown significant differences.
- Simplicity and speed of analysis allows its application for a large number of samples.

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 320 nm was correlated with the quercetin concentration by linear regression (molar extinction coefficient 23.2 (±0.3) × 103 M−1 cm−1). 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.