INTRODUCTION Sysmbrium irio (Synonym- London Rocket) belongs1 to the Family Cruciferae and commonly known as "Khubkalan". It is used as an expectorant, restorative and used externally as a stimulating poultice. The plant is indigenous to northern India and its distribution extends through Afghanistan to Europe and the Canary Islands 2,3 . The seeds have a hot sharp taste. An extensive survey of literature has revealed that the fixed oil of S.irio has been reported partly 4 . The unsaponifiable part of the oil, however still remains to be studied for its components and its pharmacological activity. Protein fraction of the seed also needs its isolation and separation. MATERIALS AND METHODS The seeds of S. irio were dried in a hot air oven at 40-50°C for 48 hrs. The dried seeds were crushed to a coarse powder in an iron pestle & mortar. The powder was sieved & stored in air light container. Biomedical & Pharmacology Journal Vol. 2(1), 183-184 (2009) Isolation and separation of proteins from the seeds of Sysmbrium irio AKHLESH SINGHAI 1 * , GOVIND NAYAK 1 , ASHOK BUDHWANI 2 and ASHISH SINGHAI 3 1 Lakshmi Narain College of Pharmacy , Bhopal (India). 2 Daksh Institute of Pharmacy, Chhatarpur (India). 3 VNS Institute of Pharmacy, Bhopal (India). (Received: March 25, 2009; Accepted: May 13 , 2009) ABSTRACT In the present investigation we have attempted the isolation and separation of proteins from the seeds of Sysmbrium irio. Its seeds are used as an expectorant, restorative and externally as stimulating poultice. The protein content was found to be 18.25%. The amino acids were identified by Thin layer and Paper Chromatography. These studies revealed the presence of amino acids, namely- Serine, Glycine, Threonine, and Methionine, Leucine, Lysine, Proline, and Glutamic acid and two remained Unidentified. Key words: Sysmbrium irio, Proteins, amino acids. Chemical Various chemicals used were potassium Iodide, Iodine, Metallic Mercury from CDH chemical. Sodium Hydroxide, Sodium Silicate, Conc. Sulfuric acid, Selenium, Copper Sulfate & Potassium Sulfate of analytical Grade were used. Isolation of Proteins The isolation of the proteins from seeds of S. irio was carried out by salt solution method. About 150 g defatted seeds of S. irio were crushed and soaked in 10%w/v Sodium Chloride solution for 10 hrs with occasional shaking. After 10 hrs maceration, the mixture was filtered & marc discarded. To the filtrate 0.5 N HCl was added slowly to get proteins precipitation (pH 3.0- 4.5).The proteins obtained were separated by filtration, washed, dried and weighed. The presence of organic nitrogen was found positive 5-7 . Determination of nitrogen content of proteins in seed 8 Estimation method consisted of two parts: (i) Conversion of nitrogen to ammonium Sulfate by Kjehldal method,