A statistical design of experiments DOE was applied to investigate biodiesel fuel BDF production process from sunflower waste cooking oil SWCO using heterogeneous bio-catalyst produced from eggshells ES. It was based on 3 level D-optimal design involving as factors methanol:oil M:O molar ratio, catalyst concentration (wt%), reaction time (min) and mixing rate (rpm). Twenty runs were carried out. A predictive linear interaction model has been correlated finding out how significant the effects of

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... these variables are in practice. LINGO software was used to find out the optimum values of the aforementioned variables for enhancing the process. According to the results obtained, the most dominant positive factor influencing the response variable (% BDF yield) was M:O molar ratio followed by catalyst concentration (wt%) and mixing rate in a decreasing order while the reaction time showed to have a negative effect on the yield. The maximum BDF yield (98.8% and 97.5%, predicted and experimental, respectively) was obtained at M:O 6:1 M ratio, catalyst concentration 3 wt%, reaction time 30 min, mixing rate 350 rpm and 60°C. Also response surface methodology RSM has been applied to study the interactive effects of independent variables on BDF yield. It was found that, the interaction between M:O and catalyst concentration (wt%) has more significant effect than interaction between other variables. The activity of the produced bio-catalyst was comparable to that of chemical CaO and immobilized enzyme Novozym 435. All the physicochemical characteristics of the produced BDF using the prepared bio-catalyst and its blends with petro-diesel fuel PDF are completely acceptable and meet most of the required standard specifications.