Design of Experiments and Optimization of Monacolin K Green Extraction from Red Yeast Rice by Ultra-High-Performance Liquid Chromatography

Monacolin K (MK), in red yeast rice (RYR) in the forms of lactone (LMK) and hydroxy acid (AMK), is known for its anti-hypercholesterolemic activity. Under the rising demand for natural bioactive molecules, we present a green ultrasound-assisted extraction (UAE) optimization study for MK in RYR. The development and validation of a fast, sensitive, selective, reproducible, and accurate ultra-high-performance liquid chromatography (UHPLC) method coupled to diode array detection for LMK and AMK allowed us to evaluate the MK recovery in different extract media. Firstly, the ethanol comparability to acetonitrile was assessed (recovery of 80.7 +/- 0.1% for ethanol and 85.5 +/- 0.2% for acetonitrile). Then, water/ethanol mixtures, with decreasing percentages of organic solvent, were tested by modulating temperature and extraction times. Water extractions at 80 degrees C for 10 min produced MK yield > 85%. Thus, UAE conditions were optimized by a DOE study using a water-based formulation (mouthwash). The optimal total MK extraction yield (86.6 +/- 0.4%) was found under the following conditions: 80 degrees C, 45 min, 5 mg mL(-1) (RYR powder/solvent). Therefore, the new single-process green approach allowed the simultaneous direct extraction of MK and mouthwash enrichment (MK concentration = 130.0 +/- 0.6 mu g mL(-1)), which might be tested for the prevention and treatment of periodontitis or oral candidiasis.