The broken rice was suspended in distilled water at a solid/solvent ratio of 1:20 (w/v), and a preliminary wet mill, called colloid milling, was performed for 30 min using Labor-pilot 2000/4 colloid mill (IKA Co., Germany). Microfluidization was then done twice by forcing milled slurry through a narrow orifice at a pressure of 100 MPa using a microfluidizer (Microfluidics Co., USA). The treated slurry was centrifuged at 8000 g for 10 min in a CR22G centrifuge (Hitachi Co., Japan), and the supernatant was decanted. Due to density differences between protein and starch, two different layers in the precipitates were observed, corresponding to protein-rich and carbohydrate-rich fractions, respectively. The supernatant, protein-rich fractions, was carefully scraped with a spatula and redispersed with 10 times distilled water (w/v).