The aim of this paper was separation and purification of Lactic acid (LA) produced from Sisal Waste by investigating the influence of modified hybrid short path evaporation (M-HSPE) system on the concentration of LA. A 22 full factorial design was used to study the influence of temperature and time on the concentration of LA and % mass obtained in distillate and residue flask. Minitab V.17 was used in designing the LA concentration experiments and in calculating the effect of each variable and their interactions. Higher LA concentration of 108 g/L occurred at system temperature of 47°C for 25 minutes. This represented a concentration of 4.5 times the initial concentration (24 g/L) with 100% recovery in residues stream/ flask. Therefore, the M-HSPE system was able to concentrate LA to almost twice the original hybrid short path evaporation (HSPE) system. According to ANOVA, LA concentration presented a correlation coefficient of 0.993, which confirm that, linear model correlated well with the experimental data. The system temperature (Temp.), operation time (time) and the interaction between system temperature and time were statistically significant variables for LA concentration at alpha = 0.05.The financial support by Tanzania commission for Science and Technology (COSTECH) under Bioplastics Project and DAAD- in Country scholarship through University of Dar es Salaam (UDSM)-Tanzania.