Pharmaceutical Study Of Compound Kuxuanshen Zhili Granules

Objective:Compound Kuxuanshen Zhiying Granules is based on Picria fel-terrae as a medicine,supplemented by Ilex rotunda,Rhodomyrtus tomentosa,and Artemisia anomala.It has the functions of clearing away heat and detoxifying,astringent and anti-diarrheal,and dampness and pain relief.Clinically,which is used for chicken damp heat diarrhea syndrome.This study aims to carry out the pharmaceutical research of Compound Kuxuanshen Zhiying Granules in order to lay the foundation for the subsequent research and development of this new veterinary drug.Methods:（1）Artificial heat and humidity environment and the inducing factor of bacterial infection were used to establish the model of chicken damp heat diarrhea syndrome.The indicators of morbidity,mortality,necropsy and bacteriological examination were used to investigate the model’s challenge strains,inoculation route and inoculation dose.The clinical symptoms,pathologic autopsy and laboratory diagnosis were used to comprehensively determine whether the model was established.（2）According to the principles of dialectic treatment,law,prescription,and medicine of Chinese veterinarians,Picria fel-terrae is the king medicine,Ilex rotunda and Rhodomyrtus tomentosa are the common medicine,and Artemisia anomala is the adjuvant medicine.Applicable to damp-heat diarrhea syndrome,the study established the treatment principles of"clearing heat and detoxifying,astringent and antidiarrheal,and dampness and analgesic"and also formulated three different prescriptions and conducted clinical efficacy trials,and finally comprehensively evaluated the effective formula.Furthermore,based on the effective formula,the study adopted the uniform design for prescription proportioning test and carried out mathematical analysis of uniform design according to the change rate and protection rate of the CRP,ALT and AST tests before and after which are the main pharmacy inspection indicators.Next,the study investigated the ratio of different prescriptions and screen out the best prescription based on the diarrhea,mortality,weight gain rate and liver disease rate which was used as an auxiliary evaluation index of drug efficacy.（3）Investigating the extraction process of Compound Kuxuanshen Zhiying Granules extract powder with different extraction solvents;taking medicine auxiliary ratio,sucrose dextrin ratio,ethanol concentration as the main investigation factors;taking comprehensive scores of moisture absorption rate,melting rate and molding rate as the investigating index;taking the advantages of the L₉（3⁴）orthogonal table to optimize the molding process of Compound Kuxuanshen Zhiying Granules;and the critical relative humidity of the finished particles was investigated.（4）The TLC was adopted to make a qualitative analysis of Picria fel-terrae,Ilex rotunda,Rhodomyrtus tomentosa,Artemisia anomala in Compound Kuxuanshen Zhiying Granules.Also,the HPLC was used to assay the Picfeltarraenin IA in Compound Kuxuanshen Zhiying Granules.Results:（1）The model of chicken damp heat diarrhea syndrome was successfully established and the modeling conditions were as follows:ambient temperature 42±2℃;relative humidity 85±2%;14-day-old liangfenghua chicken;the E.coli standard strain CVCC1555 was used as the inoculation strain;the inoculation route was intraperitoneal injection;the concentration of bacterial solution was 1×10¹¹CFU/m L and the inoculation volume was 0.5 m L/head.（2）The best prescription contains Picria fel-terrae 15 g,Ilex rotunda 30 g,Rhodomyrtus tomentosa 26 g,and Artemisia anomala 6 g.（3）The best extraction process are as follows are extract the Picria fel-terrae separately by using alkaline solution（p H=11）.Combine the Rhodomyrtus tomentosa,Artemisia anomala,Ilex rotunda three flavored medicinal water and then extract 3 times with the material liquid ratio of 1:10（w/v）,1 h each time.Concentrate,centrifuge and spray dry to obtain extract powder,mix the extract powder in proportion to make compound Picria fel-terrae extract powder.（4）The best forming process is Extract powder including excipient=1:7,sucrose:dextrin=4:1,ethanol concentration as 70%.The critical relative humidity of the finished particles is75.98%.（5）The results of TLC indicated that Picria fel-terrae,Ilex rotunda,Rhodomyrtus tomentosa,and Artemisia anomala in Compound Kuxuanshen Zhiying Granules showed consistent spots at the same position as their corresponding control herbs.（6）HPLC results indicated that:Picfeltarraenin IA raging 21.48-107.40μg/m L has a good linear relationship（R²=0.9998）,the average sample recovery is 92.50%,RSD=1.90%（n=6）.Conclusion:（1）The pathological model of artificially infected chicken with heat and diarrhea syndrome has been successfully established in the research.The model is also well restored and stable for a long time.（2）The best prescription of Compound Kuxuanshen Zhiying Granules has been successfully selected.（3）The preparation process of Compound Kuxuanshen Zhiying Granules has been successfully optimized.（4）The established qualitative and quantitative analysis method is easy to operate and the results are accurate and reliable,which can be used for the identification of Picria fel-terrae,Ilex rotunda,Rhodomyrtus tomentosa,Artemisia anomala in Compound Kuxuanshen Zhiying Granules as well as for the content determination of Picfeltarraenin IA.