Application of Box-Behnken Design for Ultrasound- Assisted Extraction and Recycling Preparative HPLC for Isolation of Anthraquinones from Cassia singueana

Abstract

Introduction – Cassia singueana Del. (Fabaceae) is a rare medicinal plant used in the traditional medicine preparations to treat various ailments. The root of C. singueana is a rich source of anthraquinones that possess anticancer, antibacterial and antifungal properties. Objective – The objective of this study was to develop ultrasound-assisted extraction (UAE) method for achieving a high extraction yield of anthraquinones using the response surface methodology (RSM), Box-Behnken Design (BBD), and a recycling preparative HPLC protocol for isolation of anthraquinones from C. singueana. Methodology – Optimisation of UAE was performed using the Box-Behnken experimental design. Recycling preparative HPLC was employed to isolate anthraquinones from the root extract of C. singueana. Results - The BBD was well-described by a quadratic polynomial model (R2 = 0.9751). The predicted optimal UAE conditions for a high extraction yield were obtained at: extraction time 25.00 min, temperature 50C and solvent-sample ratio of 10 mL/g. Under the predicted conditions, the experimental value (1.65 ± 0.07%) closely agreed to the predicted yield (1.64%). The obtained crude extract of C. singueana root was subsequently purified to afford eight anthraquinones. Conclusion - The extraction protocol described here is suitable for large-scale extraction of anthraquinones from plant extracts