Optimization of Microwave-Assisted Extraction for the Determination of Glycyrrhizin in Menthazin Herbal Drug by Experimental Design Methodology

Chromatographia. 2009;70(1):191-197. doi: 10.1365/s10337-009-1146-4. Epub 2009 Jun 16.

Abstract

In the present study, a microwave-assisted extraction (MAE) method has been investigated for the extraction of glycyrrhizin from Menthazin herbal drug. The extracted samples have been analyzed by a developed reversed-phase liquid chromatography with ultraviolet detection. The separation was performed by a Eurospher-100 C8 reversed-phase column (250 × 4.6 mm i.d., 5 μm) and the mobile phase consisted of methanol:acetonitrile:water:glacial acetic acid (30:30:40:1 v/v/v/v) with a flow rate of 0.8 mL min-1. The extraction procedure has been screened by a two level full factorial design for determination of statistically significant parameters. Thereafter, the identified parameters, extraction temperature, time and solvent volume were optimized by a Box-Behnken design. The proposed mathematical model was based on analysis of variance results and correctly explained the behavior of the response in the experimental domain. R 2 value adjusted for numbers of degrees of freedom was 0.9915 and P-value for lack of fit, 0.8499 at the 95% confidence level, P > 0.05. The optimal condition identified were extraction temperature, 70 °C, time, 13.8 min and solvent volume 2.0 mL. To evaluate the applicability of the proposed MAE method, results were compared with those obtained with the liquid extraction method. Extraction efficiency and precision were higher when MAE has been used. The proposed method allows extracting the glycyrrhizin in a small quantity of solvent and faster than the liquid extraction method.

Keywords: Box–Behnken design; Column liquid chromatography; Experimental design; Glycyrrhizin; Microwave-assisted extraction.