H-intervals was selected. Figure 3. Time course in the conversion of FAME when methanol was added at distinct intervals. The reaction was carried out at 40 for 72 h with water TLR7 Antagonist Accession content of 10 (w/w of oil). The molar ratio of methanol to oil was 3:1; 3 separate additions at 0, 12 and 24 h () or at 0, 24 and 48 h (), 1 third from the total quantity every time.60 50 Conversion ( ) 40 30 20 1040 Reaction time (h)2.3. Model Fitting and Evaluation of Variance In addition to temperature and volume of methanol, the volume of water is also crucial for the synthesis of FAME. Lipase μ Opioid Receptor/MOR Agonist drug possesses the exclusive feature of acting at the interface among an organic and an aqueous phase. The addition of water facilitates the formation of interfacial location; nonetheless, excess water could stimulate competitive hydrolysis reactions [7]. The optimal water content is really a compromise amongst minimizing hydrolysis and maximizing enzyme activity for the transesterification reaction [15]. Determined by previous reports employing P. cepacia lipase immobilized on various supports for the transesterification of triglyceride to biodiesel [15,32,33], the variables chosen for optimization and the corresponding ranges had been temperature from 35 to 50 , water content material of 1 to 20 (w/w of oil), as well as the molar ratio of methanol to oil from 3:1 to 8:1. The style of experiments as well as the corresponding data are provided in Table 1. Just after fitting the data with many models followed by analysis of variance (ANOVA), the following quadratic polynomial most suitably described the correlation among conversion as well as the tested variables: Y = -347.13 + 13.1A + 33.48B + eight.38C 0.16A2 three.31B2 0.43C2 + 0.06AB + 0.07AC 0.1BC (2)exactly where Y, A, B and C were conversion of FAME, temperature, substrate molar ratio (methanol/oil), and water content material ( , w/w of oil), respectively. The F-value of 25.92 for the model was higher than F0.01,9,7 of six.72, indicating the model was important at confidence degree of 99 . The F-value for lack of fitInt. J. Mol. Sci. 2013,was 6.42, substantially reduced than F0.01,three,four of 16.69, indicating lack of match was insignificant. General, the model had a tiny p-value of 0.0001 as well as a suitable coefficient of determination (R2 = 0.97), clearly indicating that the model was very substantial and adequate to describe the correlation amongst the conversion of FAME along with the tested variables. The higher value of adjusted determination coefficient (Adj. R2 = 0.93) also supported the significance on the model. The value of sufficient precision (a measure of signal to noise ratio) with the model was 14.29, that is higher than 4, hence giving adequate model discrimination [22]. Water content material and each of the square terms had been significant for the approach with p-values smaller than 0.05 (Table 2). Table 1. Three-level-three-factor Box-Behnken design and style of experiments and also the corresponding conversions.Remedy No. a Temperature ( ) 1 35 (-1) 2 50 (1) 3 35 (-1) four 50 (1) five 35 (-1) six 50 (1) 7 35 (-1) eight 50 (1) 9 42.five (0) ten 42.5 (0) 11 42.five (0) 12 42.5 (0) 13 42.5 (0) 14 42.5 (0) 15 42.5 (0) 16 42.five (0) 17 42.five (0)aVariable b Molar ratio (methanol/oil) Water content ( , w/w of oil) 3 (-1) 10.five (0) three (-1) 10.5 (0) eight (1) 10.five (0) 8 (1) 10.five (0) five.5 (0) 1 (-1) five.5 (0) 1 (-1) five.five (0) 20 (1) five.5 (0) 20 (1) three (-1) 1 (-1) eight (1) 1 (-1) three (-1) 20 (1) eight (1) 20 (1) 5.five (0) 10.five (0) five.5 (0) ten.five (0) 5.five (0) ten.5 (0) 5.five (0) 10.five (0) five.5 (0) ten.5 (0)Conversion ( ) 48 1 54.1 0.3 42.70 0.01 52.six 0.4 13.five 0.1 eight 44.6 0.9 58.13 0.06 14.0 0.9.