Phenolic compounds in plants function as a chemical defense system to inhibit the growth of pathogens. Folin-Ciocalteu (FC) reagent may be used to estimate the total phenolic compounds using spectrophotometry. Microanalysis of the phenolic compounds using FC reagent has not been widely used. This study aimed to develop a rapid method in estimating total phenolic compounds in soybean grain extract using a 96-wells microplate. An optimum kinetic reaction was achieved after 90 minutes of incubation. Responses of 25–1,000 μg/ml of gallic acid were linear (r = 0.99). Small amount of reagent was needed in the Microplate 2 (MP) technique. This method was more efficient, practical and environmentally friendly. Hundreds of samples could be analysed using the MP 2 in a shorter time, very useful when working with large number of soybean germplasm collection.