Abstract: Six-leaves seedling of barley as raw material, investigated the most optimum conditions of extraction of barley green.Compared to heating solvent extraction, microwave-assisted extraction and ultrasonic-assisted extraction on barley leaf exhibited more amount of barley green.Under ultrasonic treatment 30min at 40℃, followed by microwave-assisted extract 8min at 300W power, the best quality of barley green was attained, which contained total flavonoids 11224.85μg/g of green weight and had the DPPH free radicals scavenging potential of 78.52%.The nutrient components and antioxidant activity of barley green descended obviously at cold storage and room temperature.Betain and total flavonoids in barley green were easy to keep, while soluble protein, soluble total sugar and super-oxide dismutase (SOD) were hard to keep at different temperature conditions.Compared with the room temperature, the cold storage could better keep the nutrient components and antioxidant activity of barley green.