In this study the aqueous extract of alfalfa leaves was screened for identification of hypoglycemic active compound (s). For this purpose the aqueous extract of alfalfa leaves, were separated by Gel Filtration and Ion-Exchange Chromatography. Only one fraction, which was obtained from Ion-Exchange Chromatography significantly (P<0.001), lowered blood glucose level in normal mice (intraperitonealy). At dose levels of 0.5 mg/kg and 10 mg/kg body weight, there was 26.9% and 34% reductions in blood glucose levels from that of control, respectively after 2 hours of protein fraction administration.The electrophoretic assay shows that only one component is present in the hypoglycemic active protein fraction obtained from Ion-Exchange Chromatography.The M.Wt. of the active protein was about 60,000 daltons and the amino acid analysis shows that the active fraction contains 14 types of amino acids, with unusual percentage (93%), of amino acid derivative 1-methylhistidine. This protein, which contains such amino acid derivative in such high percentage, which is not discovered in other source;it is worth to test such protein further.