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Total flavonoid content of the extracts was estimated by aluminium chloride colorimetric method. After the extraction, 1?mL of extracts was diluted with 4?mL of distilled water. 5%, 0.3?mL NaNO2 solution was added to each sample. After 5?min %10, 0.4?mL of AlCl3 and then at 6?min 1.0?M 2?mL NaOH solution were added. Absorbances of the solutions were read at 475?nm. The wet wipes were prepared by allowing dry towels to absorb 6?mL of diluted extraction samples. The hand sterilizing liquid was prepared by the addition of 9?g sodium alginate to 600?mL of diluted extraction samples. In order to determine the antimicrobial activity of the extracts different dilution ratios (10 and 30?mL/L of solution) JQ1 supplier were added to the usual growth medium of mixed microorganism cultures. In the culture, there were 6 subspecies of Bacillus and 3 subspecies of Lactobacillus types present. The growth in those of extracts was analyzed by measuring their absorbance at 420?nm during 30?h. 3. Results and Discussion 3.1. Regression Model and Optimization The coded variables and their respective yields were entered into the software (Table 2). The suggested models were produced by using Box-Behnken design. Applying all the statistical tests, the reduced cubic model was found to be the best model that describes the surface of yield. The statistical significance of the model was evaluated by analysis of variance (ANOVA) as presented in Table 3. Since the larger the magnitude of the F-value and the smaller the p values, the more significant the corresponding coefficient, the most effective parameter on the extraction yield was found to be solid-to-liquid ratio. Extraction temperature E-64 was found more effective than extraction time. Table 2 The coded variables and experimental yield values in the study. Table 3 Analysis of variance (ANOVA) for the reduced cubic model. R428 In order to check the model found, the predicted and actual yield values were compared. As can be seen from Figure 1, good agreement was observed. The regression coefficients were also in agreement with this conclusion (Table 3). Thus, the empirical relation explaining the surface was found as in Yield=64.65+11.61x1?13.44x2+6.9x3?1.63x1x2+2.89x1x3+2.28x2x3?3.71x12+4.87x22+1.73x32+9.58x12x2?3.61x12x3?7.78x1x22, (4) where x1, x2, and x3 were the coded values of the parameters defined in Table 2. Response surfaces were obtained by the respective button of the software. The parameters and their interactive effects were shown in Figure 2. The increase in extraction yield was observed with decreasing the solid-to-liquid ratio (a), while increasing the temperature up to a specific value less than 50��C (b) and the extraction time as soon as possible (c). The requirement of high temperature and long time may be due to different types and amounts of antimicrobial compounds present in plant to be extracted into the solvent.