Paper-based radial chromatographic assay for rapid in situ measurement of gelatinized starch concentration

Starch gelatinization is pivotal in food processing, impacting viscosity, texture, and functionality. However, current methods for determining concentration and viscosity of gelatinized starch (GS), such as falling ball and rotational viscometers, are impractical for in situ measurements due to bulky equipment. Here, we introduce a paper-based radial chromatographic assay (PRCA) method for rapidly measuring the concentration of gelatinized starch (GS). We utilized Evans Blue (EB) as a chromatic agent to label GS and generate color-ring patterns on a nitrocellulose membrane based on the “Coffee-ring” effect. The resulting PRCA patterns varied depending on the GS concentrations, yielding a good linear correlation (R² > 0.967) between grayscale value and viscosity. This method enables rapid measurement of GS concentration within 10 s, even with varying ratios of amylose to amylopectin. PRCA demonstrates a detection limit for GS concentration between 0.027 % and 0.051 %, which is 10-20 fold lower than that of traditional rotational-viscometer. The intensity of PRCA patterns at pH 3 (45.8) was significantly lower than those at higher pH levels (5 to 9), consistent with the decreased viscosity measured by the viscosimeter. The intensity of PRCA patterns did not change with increasing temperature from 25°C to 90°C, despite the significant decrease in viscosity from 345 mPa•s to 195 mPa•s. Additionally, PRCA can monitor changes in starch viscosity during the hydrolysis process, which decreases from 744 mPa•s to 6 mPa•s. This capability provides an efficient means of measuring the concentration and viscosity of GS during food processing.