The study observed the biodiesel separation of glycerine for several concentration. In this study, the observations of the average droplet velocity were carried out for biodiesel glycerine mixture with concentrations of 0.1, 0.2, 0.3, 0.4 and 0.5. The biodiesel glycerine mixture was put in measuring cylinders with diameters of 1.3 cm, 2.8 cm 3.9 cm. The Biodiesel and glycerine mixture was stirred using a magnetic stirrer with a rotational speed of 1000 rpm for 1 h. The separation time was observed for each mixture. The droplet diameter and average speed of biodiesel can be determined by using the Stoke’s equation. The viscosity of glycerine is 0.064 Pa.s, whereas the density of glycerine and biodiesel respectively 1200 kg/m3 and 880 kg/m3. The Hunter’s equation can be applied for 1,3 cm measuring cylinder diameter with the value Фc and k respectively 0.6 and 6.12, and the Hunter equation is not valid for the measuring cylinder diameter of 2,8 and 3,9 cm.