Studying On The Changes Of The Main Compounds In Co-boiling Extracts, Pooled Individual Boiling Extracts In Dioscorea Bulbifera L.and Glycyrrhiza Uralensis Fisch. Decoction

Objective:1. To establish the analysis methods liquid chromatography fingerprinting on Dioscorea bulbifera L. and Glycyrrhiza uralensis Fisch.To compare the contents of the seperated and mixed decoction of Dioscorea bulbifera L. and Glycyrrhiza uralensis Fisch. Investigate the theory of Glycyrrhiza uralensis Fisch. reduce the liver toxicity of Dioscorea bulbifera L. and provide the experimental basis for Dioscorea bulbifera L. used safely and resonable in clinical,at the same time,we explore the detoxification mechanism of the traditional antidote Glycyrrhiza uralensis Fisch.2. To establish the analysis methods liquid chromatography on Diosbulbin B and Glycyrrhizin.To compare the contents of Diosbulbin B and Glycyrrhizin in the seperated and mixed decoction of Dioscorea bulbifera L. and Glycyrrhiza uralensis Fisch.Methods:1. Part 1:To compare the contents of the seperated and mixed decoction of Dioscorea bulbifera L. and Glycyrrhiza uralensis Fisch. The samples were separated at 30℃on a PLATISILTM-C18 column (4.6 mm×250 mm,5μm)and gradient mobile phase were used(A:MEOH,B:0.1%HAC).The elution gradients included 0-5 min(2-17% A),5-15 min(17-40% A),15-40 min(40-80%A),40-65 min(80-100% A),65-70 min(100-100%A).Flow rate was 0.8 mL-min-1 and the detection wavelength was set at 260 nm.2. Part 2:To compare the toxicity and effects of contents of Diosbulbin B in the seperated and mixed decoction of Dioscorea bulbifera L. and Glycyrrhiza uralensis Fisch.The samples were separated at 25℃on a PLATISILTM- C18 column(4.6 mm×250 mm,5μm) eluted with acetonitrile-water-glacialacetic acid (40:60:0.1).Flow rate was 0.8 mL-min-1 and the detection wavelength was set at 210 nm.3. Part 3:To compare the contents of Glycyrrhizin in the seperated and mixed decoction of Dioscorea bulbifera L. and Glycyrrhiza uralensis Fisch. The samples were separated at 25℃on a PLATISILTM-C18 column(4.6 mm×250 mm,5μm) eluted with acetonitrile-water-glacialacetic acid (40:60:3).Flow rate was 0.8 mL-min-1 and the detection wavelength was set at 250 nm.Result:1. We established the method of liquid chromatography fingerprinting of the seperated and mixed decoction of Dioscorea bulbifera L. and Glycyrrhiza uralensis Fisch. The main compounds was no difference between the two solution.But the content of the some chemical substances has made great changes.2. In the range of 37.5-310μg·mL-1 (r=0.9999),the linear relationship were excellent;th mean recoveries were between 100.89% and 100.55%,and RSD values were 3.26% and 2.79%. The content of Diosbulbin B:Dioscorea bulbifera L. (1.175 mg·g-1)> seperated decoction (0.808 mg·g-1) > mixed decoction (0.717 mg·g-1)。3. In the range of 30.2-604.0μg-mL-1 (r=0.9999),the linear relationship were excellent;the mean recoveries were between 99.82% and 99.85%,and RSD values wer 2.78% and 3.48%. The content of Glycyrrhizin:Glycyrrhiza uralensis Fisch. (10.29 mg·g-1)> seperated decoction (7.245 mg·g-1)> mixed decoction (4.322 mg·g-1)Conclusion:1. These methods are simple,high reproducibility,specificity,reliablity,can determinate th the contents of Diosbulbin B and Glycyrrhizin.2. The main chemical substances were no difference between the two solution.But the content of the main chemical substances had made great changes.3. The contents of Diosbulbin B and Glycyrrhizin reduced.The reducation of Diosbulbin B maybe the reason for the toxicity of Dioscorea bulbifera L reduced. We will mak a further investigation of the specific reason.