Studies On Toxicity Test In Vitro And Capillary Electrophoresis Of Chinese Medicine

As more and more research have recived about traditional Chinese medicine efficacy,their effective constituent ,molecular structure,the pharmacodynamics action,toxicology and so on have also studied partily.But much Chinese medicine havn,t estimated by strict cytotoxicity test and hazardous analysis because of the restriction of historical condition. We have learned more toxicity from clinical manifestation while less about essence and action mechanism; more about acute toxicity that found in short-time while less about the potential toxicity of organic injury. With the development of genetic toxicology,a mount of research findings about Chinese medicine genetic toxicology have been made,and they provides a scientific basis for safety application of Chinese medicine.For this study, the human hepatocyte carcinoma cells.HepG2 was selected as test system for the following experiments. Actute toxicity tests and genetic toxicity tests were studied by neutral red(NR) stainied and the single cell gel electrophoresis(SCGE) respectively,and then the second screening of effective ingredient in Chinese herbal medicine have been made by high performance capillary electrophoresis (HPCE) after toxicity tests, it have provided experiment materials for the safety of chinese clinical medication.1 To use NR analyse dazens of Chinese medicines which may cause genetic toxicity for HepG2 cells,and then assessed the genetic toxicity by SCGE on two of them . 2 With Ultrasonic extraction technology,the optimum extraction condition of total isoflavone from Rhizoma Belamcandae and total alkaloids (MTA) in Evodia rutaecarpa was confirmed by the orthogonal test: extracting 120 min by 75 mL methanol,after marinated 12h,under these conditions was 37.7 mg/g, while extracting 120 min by 200 mL methanol,after marinated 2h,under 100W ultrasonic frequency, the extraction rate of MTA in these conditions was 2.99%.3 Separation of Tectoridin and Tectorigenin by High Performance Capillary Electrophoresis. Effects of several important factors were investigated such as the pH and concentration of running buffer, operation voltage and injection time. Tectoridin and Tectorigenin can be separated under the optimized conditions: 20.0 mmol/L borate(pH=9.0) as running buffer ,15 kv operation voltage, 6s as injection time.4 A quick and simple method of High Performance Capillary Electrophoresis with amperometrie detection (CE/AD) was established for the detection of Rutecarpine in Evodia Rutaecarpa. Effects of several important factors were investigated such as the pH and concentration of running buffer, operation voltage, injection time. Rutecarpine can be detected in ten minutes under the optimized condition: 5 mmol/L borate (pH = 9.5) as running buffer, 15 kv operation voltage. The concentration of Rutecarpine was lined with the peak height in the range of 1.74×10-⁸～3.48×10^-6 mol/L, Y = 3.0×10^-7+1.2×10^-3·X ,r=0.9996. Recovery rate between 94.0%～106.0%. This method has been successfully applied for Evodia Rutaecarpa sample with satisfactory results.5 Rutecarpine from Evodia rutaecarpa(Juss.)Benth. was determined by Capillary Electrophoresis with Conductivity detection. Effects of several important factors to determine Rutecarpine from Evodia rutaecarpa(Juss.)Benth. were investigated such as running buffer and its concentration, the pH ,operation voltage , the injection height and injection time. Under the optimized condition: 4mmol/Lborate (pH8.9) as running buffer, 10 kv operation voltage, the injection height was 20cm,the average recovery rate between 93.4%～102.4%.