Scanometry as microplate reader for high throughput method based on DPPH dry reagent for antioxidant assay

Abstract

The stable chromogenic radical 1,1′-diphenyl-2-picrylhydrazyl (DPPH) solution was immobilized on the microwell plate as dry reagent to construct a simple antioxidant sensor. Then, a regular flatbed scanner was used as microplate reader to obtain analytical parameters for antioxidant assay using one-shot optical sensors as scanometry technique. Variables affecting the acquisition of the images were optimized and the analytical parameters are obtained from an area of the sensing zone inside microwell using the average luminosity of the sensing zone captured as the mean of red, green, and blue (RGB) value using ImageJ® program. By using this RGB value as sensor response, it is possible to determine antioxidant capacity in the range 1–25ppm as gallic acid equivalent (GAE) with the response time of 9min. The reproducibility of sensor was good (RSD<1%) with recovery at 93%–96%. The antioxidant sensor was applied to the plant extracts, such as sappan wood and Turmeric Rhizome. The results are good when compared to the same procedure using a UV/Vis spectrophotometer.