Optimized Extraction Method for Kleeb Bua Daeng Formula with the Aid of the Experimental Design

Kleeb Bua Daeng formula is one of the popular remedies sold in Chao Phya Abhaibhubejhr Hospital, Thailand. This formula contains Piper nigrum L., Nelumbo nucifera Gaertn., and Centella asiatica L. as active components. Owing to getting the highest content of its phytochemical compounds, the conditions of solvent extraction for this formula were optimized. The type of solvent, number of extraction times, and ratio between the material and solvent were varied in this study using the Box–Behnken design. The important phytochemical constituents (total phenolics, flavonoids, carotenoids, and anthocyanins) were also determined. From the result of this study, it was found that the highest content of each total active compound was obtained from different conditions such as the optimal extraction condition of phenolic content was obtained using methanol as solvent, one time of extraction, and the ratio of powder and solvent was 1 : 6. Thus, the variation of solvent extraction condition could affect the phytochemical content. Further studies about the herbal formula involving the extraction process should concern the variation of extraction conditions to get the highest content of the active compound.