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Kaempferol (KA) is a natural flavonol that can be found in plants and plant-derived foods with a plethora of different pharmacological properties. In the current study, we developed an efficient extraction method for the isolation of KA from ultrasonicated basil leaves ( Ocimum basilicum ). We successfully employed a Box-Behnken design (BBD) in order to investigate the effect of different extraction variables including methanol concentration (40-80%), extraction temperature (40-60 °C), and extraction time (5-15 min). The quantification of KA yield was carried out by employing a validated densitometric high performance thin layer chromatography in connection with ultraviolet detection (HPTLC-VIS). The obtained data showed that the quadratic polynomial model (R 2 = 0.98) was the most appropriate. The optimized ultrasonic extraction yielded 94.7 ng/spot of KA when using methanol (79.99%) at 60 °C for 5 min. When using toluene-ethyl acetate-formic acid (70:30:1 v / v / v ) as a solvent, KA was detected in basil leaves at an Retention factor (Rf) value of 0.26 at 330 nm. Notably, the analytical method was successfully validated with a linear regression of R 2 = 0.99, which reflected a good linear relationship. The developed HPTLC-VIS method in this study was precise, accurate, and robust due to the lower obtained results from both the percent relative standard deviation (%RSD) and SEM of the O. basilicum. The antioxidant activity of KA (half maximal inhibitory concentration (IC 50 ) = 0.68 μg/mL) was higher than that of the reference ascorbic acid (IC 50 = 0.79 μg/mL) and butylated hydroxytoluene (BHT) (IC 50 = 0.88 μg/mL). The development of economical and efficient techniques is very important for the extraction and quantification of important pharmaceutical compounds such as KA.

Optimization of Ultrasonicated Kaempferol Extraction from Ocimum basilicum Using a Box–Behnken Design and Its Densitometric Validation