Development of a Spectrophotometric Flow-Based System for the Determination of Total Polyphenol Content in Legume...

Conventional nutritional characterization of legume flours comprises costly and laborious analytical methods for nutrient quantification that establish the quality of seeds, including their macronutrient quantification, fiber, mineral, antioxidant content, and more. The quantification of total polyphenol content (TPC) in legumes is performed using different analytic methods, namely the Folin-Ciocalteu (FC) method, which is lengthy and employs potentially toxic reagents. Additionally, it is time-consuming and prone to human error if carried out in a conventional batch mode.

To develop a semi-automatic method for a faster, greener, and more precise TPC quantification, resorting to flow injection analysis (FIA).

The development of the FIA method was based on the FC method. The flow manifold was structured by performing an array of preliminary studies, which led to the establishment of the method that displayed the highest sensitivity (highest slope of the calibration curve). The method was applied to determine the TPC in various legume flours.

This novel method resulted in a throughput of 1 analysis per minute, allowing to analyze 20 samples in triplicate within an hour, with a LOD of 4.32 mg L^-1, a LOQ of 14.4 mg L^-1, and an RSD of 4.4%. The results calculated from the proposed FIA method agreed with those of the reference procedure.

By using the FIA system, a lower consumption of reagents was observed. Additionally, as there is no need to reach chemical equilibrium, a high throughput has been achieved, resulting in a faster and greener method for the determination of polyphenols in various types of legumes.