Diabetes mellitus (DM) is a metabolic disease characterized by an increase in blood glucose level above normal (hyperglycemia). Natural ingredients containing hypoglycemic substances can be used an alternative DM treatment, one of which is bitter leaf (Andrographis paniculata). Sambiloto leaves contain compositions of andrographolide and flavonoids which have anti-diabetic activity. The research method begins with simplicia preparation which begins with fermentation using Aspergillus niger with a fermentation temperature variation of 26°C, 30°C, 34°C, and 37°C. The sample is then extracted with a frequency of 42 kHz with 70% ethanol. The extraction results were used in the anti-diabetes test by comparing samples given α-glucosidase enzymes with positive control (diabetes medication) and negative control (ethanol 70% with corresponding concentration), and samples without fermentation process with coarse variations of crude extract 100 ppm, 200 ppm, 300 ppm, 500 ppm, and 5 mg/ml and added absorbance using UV-Vis spectrophotometry. Chemical components analysis was carried out using GC-MS.