Study On Extraction,purification And Hepatoprotective Activity Of Lignans In Schisandra Chinensis

Schisandra chinensis is the dried fruit of Schisandra chinensis of Magnoliaceae or Schisandra chinensis of Huazhong.It has the functions of protecting the central nervous system,protecting the liver and kidney,scavenging free radicals in the body,antitussive,expectorant,anti-tumor and anti-HIV.Lignans are the main pharmacological active components in Schisandra chinensis.In this study,the extraction,separation,purification and hepatoprotective activity of lignans were investigated.The main research contents are as follows:（1）In this study,a magnetic stirring assisted supercritical CO₂extraction method was established and applied to the extraction of eight lignans from the fruit of Schisandra chinensis.The target analytes were separated and determined by high performance liquid chromatography（HPLC）.In this study,supercritical CO₂was used as extraction solvent,the sample was pre-impregnated with ethanol solution,and the contact between samples and supercritical CO₂was promoted by magnetic stirring to improve mass transfer efficiency.In this study,various factors affecting the extraction efficiency of lignans were optimized,and compared with the traditional methods.The experimental results showed that the good linear relationship for analytes was obtained in the linear range（r>0.9998）.The limit of detection and limit of quantification were 21.30～990.20 ng/m L and 60.80～2992.80 ng/m L,respectively.The spiked recoveries were between 75.00%and 97.78%.The analysis results were satisfactory.Compared with other methods,the extraction yield obtained by this method was significantly higher than that obtained by microwave extraction and thermal reflux extraction,and comparable to that obtained by the pharmacopoeia method.This method has the advantages of large sample handling capacity,simple operation,environmental protection and high extraction efficiency,and certain potential in industrial production and application.It can be used for the extraction and separation of lignans in Traditional Chinese medicine.（2）In order to remove the impurities in the extracts and obtain high purity lignans compounds,the macroporous adsorption resin was used to separate and purify the lignans in Schisandra chinensis.In the static adsorption-desorption experiment,AB-8 macroporous resin was selected as the adsorption and separation material.When the concentration of ethanol was 30%and the adsorption time was 50 min,the optimal adsorption rate was 79.2%.When the concentration of desorption solvent was95%and the desorption time was 50 min,the optimal desorption rate was 76.1%.In the dynamic adsorption-desorption experiment,when the volume of loading solvent was 40 m L and the flow rate of adsorption was 5 BV/h,the optimal adsorption rate was 66.8%.When the volume of desorption solvent was 40 m L and the elution flow rate was 3 BV/h,the optimal desorption rate was 56.7%.The results showed that the macroporous adsorption resin method has a very good purification effect,which can be used to improve the purity of lignans in Schisandra chinensis.This study can be used as a reference for the large-scale separation and purification of lignans in industry.（3）In order to further determine the protective effect of lignans in Schisandra chinensis on liver injury,the hepatoprotective activity of the extracted lignans was studied in this study.Male mice of Kunming species were randomly divided into 6groups with 11 mice in each group,which were control group,model group,positive drug group,high dose group,medium dose group and low dose group,respectively.Control group and model group were intragastrically given sodium carboxymethyl cellulose（10 m L/kg,0.5%）.The positive control group was intragastrically given the contents of silybin capsule（calculated by silybin,86.4 mg/kg）.The intragastric doses of the high-dose,medium-dose and low-dose groups were 2.46 g/kg,1.23 g/kg and0.615 g/kg,respectively.The drug was given by gavage once a day for 15 consecutive days.After last administration at the 1h,except for the control group,the other groups were intraperitoneally injected with CCl₄peanut oil solution（0.1%,10 m L/kg）and fasted without water.After 16 h,blood was collected from the eye vein and centrifuged to determine the activity of aspartate aminotransferase（AST）and alanine aminotransferase（ALT）in serum.The results showed that,except the control group,the contents of AST and ALT were increased due to the injection of CCl₄,while the contents of AST and ALT were significantly decreased with the injection of lignans.Pathological results showed that animal liver cells can be effectively protected by high,medium and low doses of lignans from Schisandra chinensis,and liver cell damage can be alleviated to a certain extent.