Determination of Ascorbic Acid (Vitamin C) with Paper Based Colorimetric Method

Abstract

Ascorbic acid (AA) is the most important water-soluble organic biomolecule with antioxidant properties, known as vitamin C in many foods and beverages. The number of different methods developed for the determination of antioxidant properties of foods is increasing. Paper-based microfluidic analytical devices (µPADs) have been applied in the development of fast, simple, and cost-effective field analyzes in recent years. These developed analyzes are used as an alternative to laboratory-equipped qualitative and quantitative analyzes. In this study, a paper-based colorimetric method was applied for the determination of ascorbic acid. Geometric shapes with hydrophobic barrier were created on Whatman filter papers with the help of a wax pen. Using the principle of reduction of the AgNO3 solution in the presence of antioxidants in these shapes, the images of the color transformations were recorded with the help of a smartphone camera. ImageJ software was used in the image processing to determine the concentrations of standard AA solutions. Linear calibration curves for AA concentrations in the 0,2 to 1,0 mM range were obtained in grayscale and RGB (Red-Green-Blue) values. The coefficient of determination of the calibration curves (R2), respectively; 0,9389 for grayscale, 0,9728, 0,9628, and 0,8859 for Red, Green, and Blue channels. High-Performance Liquid Chromatography (HPLC) was used as a standard method to compare with the paper-based method. Freshly squeezed orange juice was used for the determination of ascorbic acid in the real food sample. The ascorbic acid value determined in orange juices by HPLC and colorimetric methods were statistically compared.