S. platensis is a microalga that contains carbohydrate composition of 30.21% which makes it potential to be used as raw material for ethanol production. Hydrolysis of S. platensis is the first step for converting its carbohydrates into monosaccharides. The second step is fermentation of monosaccharides into ethanol. This research aims to study the effect of temperature and microalgae concentration on the hydrolysis of S. platensis using sulfuric acid as catalyst. This research was conducted using 300 mL sulfuric acid of 2 mol/L, hydrolysis temperatures of 70, 80 and 90 °C, and microalgae concentrations of 20, 26.7, and 33.3 g/L. The effect of temperature is significant in the hydrolysis of S. platensis using sulfuric acid. At microalgae concentration of 20 g/L and hydrolysis time of 35 minutes, the higher the temperatures (70, 80, and 90 °C), the more the glucose yields would be (8.9, 13.5, and 22.9%). This temperature effect got stronger when the hydrolysis was running for 15 minutes. Every time the hydrolysis temperature increased by 10 °C, the glucose yield increased by 13.0% at microalgae concentration of 33.3 g/L. At temperature of 90 °C and time of 35 minutes, the higher the microalgae concentrations (20, 26.7, and 33.3 g/L), the higher the glucose yields would be (25.5, 27.7, and 28.2%). The highest glucose concentration obtained was 2.82 g/L at microalgae concentration of 33.3 g/L, temperature of 90 °C, and time of 35 minutes.