Application of Box-Behnken design for ultrasound-assisted extraction and recycling preparative HPLC for isolation of anthraquinones from Cassia singueana

Phytochem Anal. 2019 Jan;30(1):101-109. doi: 10.1002/pca.2795. Epub 2018 Oct 4.

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 an 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 high-performance liquid chromatography (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 50°C 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.

Keywords: Box-Behnken design; Cassia singueana; Fabaceae; HPLC; Optimisation; anthraquinone; ultrasound-assisted extraction.

MeSH terms

  • Anthraquinones / isolation & purification*
  • Cassia / chemistry*
  • Chemical Fractionation / methods
  • Chromatography, High Pressure Liquid / methods*
  • Models, Statistical
  • Plant Extracts / chemistry
  • Plant Roots / chemistry*
  • Proton Magnetic Resonance Spectroscopy
  • Reproducibility of Results
  • Research Design*
  • Ultrasonics / methods*

Substances

  • Anthraquinones
  • Plant Extracts