| Celandine,also known as Coptidis chinensis,is an angiosperm phylum,dicotyledonous plant class,and Papaveraceae plant.It was first recorded in "Herbal Rescue" and belongs to the whole grass class and is a perennial herb.It grows in the shady places of valleys and ravines,and is distributed in most parts of China.Contains a variety of alkaloids,known by the public for their alkaloid effects.In this experiment,celandine alkaloids were extracted and enriched,and the mechanism of action of the active components of celandine alkaloids on the immune and antioxidant functions of dogs was studied,and the safety evaluation was made.Methods:(1)Collect fresh celandine and divide it into two parts: fixed and unfinished.The fresh vegetables were used,dried in the shade,dried in the sun,and dried.The HPLC detection method was established,and the alkaloid content was verified by HPLC.The chelidrine and chelerythrine were used as evaluation indexes to discuss whether they were used dry,wet or rehydrated.(2)Celandine alkaloids were extracted by decoction,hot soaking and Soxhlet reflux extraction using the treated whole grass of celandine,and the suitable extraction method was discussed,and analyzed by single factor combined with response surface methodology.,optimize the extraction process.And the macroporous resin enrichment experiment of celandine alkaloids was carried out,11 different types of resins were screened,and more suitable resin types were found to provide the basis for the extraction and purification of the drug.(3)The clinical application of celandine alkaloids was carried out,and the maximum administration concentration and maximum administration volume were used to administer to dogs,and the health status and blood biochemical indexes of dogs were observed at 1d,3d,and 7d after administration.To make an effective evaluation of the safety,at the same time,the dogs were given 1m L/kg of celandine alkaloids,twice a day,and fed continuously for 21 days.Immunity and its effects on antioxidant function.Results:(1)By comparison,there was no significant difference in the contents of celandine and celandine in celandine before and after greening.The contents of celandine and celandine in freshly harvested celandine were the highest,respectively.It was 9.876 ± 0.625mg/g,2.321 ±0.015mg/g.After drying in the shade,the content loss of chelidrine and chelerythrine was the lowest,which were 5.643 ± 0.327mg/g and 0.924 ± 0.013mg/g,respectively.(2)Through the comparison of the three extraction methods,the decoction method has the worst effect on the dissolution of alkaloid components,and the Soxhlet extraction method has the best effect.The optimized results were that the extraction temperature was 64.86℃,the extraction time was 1.85 h,and the concentration of hydrochloric acid was 0.62%.The calculated total extraction rate of celandine alkaloids was 0.756%.The optimized results of Soxhlet extraction of celandine alkaloids were that the extraction temperature was 80.8℃,the number of siphons was 6.16,and the concentration of hydrochloric acid was 0.55%.The calculated total extraction rate of celandine alkaloids was 1.089%.(3)Through screening,LX-28 resin is more suitable for the enrichment of celandine alkaloids than other types of resin,the adsorption capacity is 4.1mg/g,the resolution rate is 76%,and under dynamic elution,80 m L is used.As the sample volume,the resin can be saturated in 160 min,and the chelidonine can appear in the 300 m L(30% ethanol)fraction,but the content is very small,and it is mainly concentrated in the 400 m L(50% ethanol)fraction to be eluted,The concentration is the highest,and chelerythrine appears in the 425 m L(75% ethanol)fraction,which is mainly eluted by 95% ethanol,and the concentration is the highest.(4)In the safety experiment on dogs,at the dose of 20 m L/kg(equivalent to 20g/kg of crude drug),there was no significant difference between the blank group and the administration group(P>0.05),and 1m L/kg At the dose of kg,chelendron alkaloids can significantly improve the body’s immunity.On the 7th,14 th,and 21 st days after administration,the levels of immunoglobulin Ig A,Ig G,and Ig M in the serum of dogs were significantly increased,and the serum inflammatory factors IL-1,The contents of IL-6 and interferon INF-γ were significantly increased,and the administration group was significantly higher than the blank control group(P<0.05),and the activity of antioxidant enzymes was also significantly increased.Compared with the blank group,the activity was significantly different(P<0.05),and the content of MDA in serum was significantly lower than that of the blank group(P<0.05).Conclusion: It is recommended to use celandine directly after harvesting.The loss of celandine alkaloids can be significantly reduced by drying in the shade.The Soxhlet extraction method uses the parameters of extraction temperature of 80℃,siphoning times of 6 times,and hydrochloric acid concentration of 0.55%.Extraction can significantly increase the alkaloid content in celandine extract.The dose of 20 m L/kg(crude drug 20g/kg)was administered to dogs,which was basically non-toxic.In addition,continuous administration of 1m L/kg twice a day for21 days can significantly improve the immune function of dogs,and enhance their disease resistance and antioxidant capacity.This experiment provides a reference for the clinical application of celandine. |
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