Optimization Of Extraction Of Effective Compounds From Trigonella Foenum-graecum L.(Fenugreek) Using Response Surface Methodology

Trigonella foenum graecum L.(Fenugreek) is an annual plant from the familyPapilionaceae-Leguminosae, and is widely cultivated in China. It was used to treat kidneydeficiency, hernia, abdominal pain, arthritis, beriberi in Chinese traditional medicine.Modern scientific research indicated that fenugreek seeds have possible properties ofhypoglycemic, antihyperlipidemic, antioxidation, anti-inflammatory and analgesic, andsteroidal saponins and polysaccharides may be the effective components in the Fenugreekseeds. So the steroidal saponins and polysaccharide in the fenugreek has a great deal ofresearch and development value.There is no effective extraction and purification method of the steroidal saponin infenugreek so far, so this paper adopted hydrophilic aqueous two-phase system (ATPS)(ethanol/K2HPO4) to purify the steroid saponins effectively. Response SurfaceMethodology (RSM) was used to design the optimization experiment, and study thevarious factors on the effect of the yield of total steroid saponins. The optimum conditionsare sample loading of3.77%, the mass concentration of K2HPO4of25.06%and ethanolconcentration of22.53%, and the maximum yield was97.93%. Under these conditions,the experimental yield was98.30%, which is well matched with the predicted model.These results show that steroidal saponins of fenugreek can be purified by aqueoustwo-phase system effectively.Studies have shown that diosgenin is its drug activity center, which is the mainnuclear of steroidal saponins in the fenugreek. Its content has a close relationship with thefurther development of drug. So we established micelle capillary electrophoresis methodto detect diosgenin. Compared with the traditional method of High Performance LiquidChromatography (HPLC), this method has the advantages of easy operation, rapid,sensitive, etc. Results showed that the mass concentration of diosgenin from the crude extraction of fenugreek is102.51mg/g, while the mass concentration of diosgenin fromtop phase of aqueous two-phase purification is395.08mg/g. It was also found thataqueous two-phase extraction plays an important role in enrichment and purification ofdiosgenin.Hot water technology is the main conventional extraction method for polysaccharidesmentioned in previous studies. This paper use ultrasonic-assisted extraction technique toextract fenugreek polysaccharides. It offers simplified manipulation, lowered energy inputand solvent consumption. A three-level, three-variable Box–Behnken design (BBD) isused to optimize the process. The optimum conditions are extraction temperature of80℃,W/M ratio of41ml/g, extraction time of50min, and the predicted maximum yield ofpolysaccharides is4.98%. Under the optimized conditions, the experimental values are inclose agreement with values predicted by the model, so this methodology could be appliedin the extraction of fenugreek polysaccharides in the natural product industry.These resultsprovide a basis for the amplification of industrialization production.