A non-catalytic method to produce biodiesel in situ from a rice milling by-product, i.e. rice bran, using subcritical water-methanol mixture has been investigated. The method was found to be unaffected by initial moisture and free fatty acids (FFA) contents in rice bran so that no pretreatment was required. The yield and purity of biodiesel were higher under CO2 atmosphere than those under N-2 atmosphere due the ability of the gas to acidify water-methanol mixture. Oil extraction from the bran was identified as the limiting step and complete oil extraction could be achieved in 3 h at 200 degrees C, 4 MPa (under CO2 atmosphere) and 43.8 wt% methanol concentration. Consequently, the highest biodiesel yield was also achieved at those operating conditions. The experimental data suggested that hydrolysis of rice bran oil into FFA followed by methyl-esterification of FFA into biodiesel could be the preferred reaction path to direct transesterification of oil. Subcritical water-methanol mixture was also able to break down complex carbohydrates in rice bran into simple sugars soluble in aqueous phase so that it could be separated easily from biodiesel. (C) 2017 Elsevier Ltd. All rights reserved.