A process for the preparation of mustard protein isolate, comprising steps such as dispersion of defatted meal in 0.1 mol/l NaCl solution, incubation, extraction at alkaline pH, followed by treatment of the protein solution with activated carbon was developed. The protein, coagulated by steam injection, was subjected to separation by centrifugation, washing and spray drying. The parameters evaluated were protein yield, purity, presence of antinutritional factors and nutritional quality of proteins. The protein yield was 58-60%. The purity of the protein isolate was 95%. The hydrolysed products of glocosinolates like isothiocyanates and oxazolidine thione levels, phenolics and phytic acid levels were low in the protein isolate. The in vitro digestibility of the protein isolate was 92.4% compared to 80.6% of the meal. Chemical score of the meal and protein isolate were similar; isoleucine was the first limiting amino acid. The calculated nutritional indices, essential amino acid index, biological value, nutritional index and C-PER of protein isolate were higher compared to meal. The protein quality as indicated by amino acid profile and PDCAAS scores for 10-12-years old and adults were 100. (c) 2005 Swiss Society of Food Science and Technology. Published by Elsevier Ltd. All rights reserved.