The Preparation Process And Quality Standard Of Jianpi Huoxue Jiedu Granules

Objective:Jianpi Huoxue Jiedu granules are based on a formula in clinical applications,which comprises eleven herbs,including radix codonopsis,rhizoma atractylodis macrocephalae,poria,fortune euonymus,radix sanguisorbae and others.In clinical practice,this formula presented good effect in treating symptoms of the patients at the remission stage of ulcerative colitis（UC）,such as diarrhea,abdominal pain,purulent stool,rectal tenesmus and so on.Since this formula was applied usually on a long-term basis,the inconvenience of decoction,preservation and portability limited its clinical application.Therefore this study followed the requirement of“Administrative Measures for the Registration of Pharmaceutical Preparations of Medical Institutions”to develop this formula in the form of granules,conducted researches on the preparation process,quality standard,stability,etc.,and prepared all the needed materials for applying for hospital preparation.Methods:This paper,based on the standard concept of decoction,conducted research on the fingerprints of water extract,intermediate and the finish product,screened for the optimal preparation process of Jianpi Huoxue Jiedu granules,established the quality standard for the granules and carried out the stability study.（1）Fingerprint study based on standard decoction:The Similarity Evaluation System Software for Chromatographic Fingerprint of Traditional Chinese Medicine（Version 2012）was utilized to establish the fingerprints of the standard decoction of the formula,water extract,intermediate and the finished product and identify the common peaks.With reference to the fingerprint of the standard decoction,the common peaks of the fingerprints of water extract,intermediate and the finished product were compared.The similarity of the fingerprints was evaluated to ensure the consistency of the product and the former formula’s ingredients.The change of the content of gallic acid as effective ingredient in these four was analyzed to evaluate if the preparation process can preserve the effective ingredient effectively.The fingerprint of the finished product was compared with the ones of the standard decoction,single herb and negative herbs to identify common peaks’attribution.（2）Preparation process study:（1）Single-factor experiment:Five factors such as crush degree,soaking time,added water amount,decocting time and frequency were investigated with extract yield rate,total polysaccharide,total flavonoids and gallic acid content as multiple indexes;Orthogonal experiment:With reference to the result of single-factor experiment,total polysaccharide,total flavonoids and gallic acid content were set as indexes to investigate crush degree,added water amount,decocting time and frequency.（2）The granulation rate and character of granules were set as indexes to screen for the appropriate type and amount of the thinner.The investigation of angle of repose determined the mesh size of the sieve for granulation.By investigating character of granules and water content,the drying time and drying temperature were determined.（3）Quality standard:Guided by the requirement of general granules in Chinese Pharmacopoeia 2015 edition,the quality of finished product was examined;Rhizoma atractylodis macrocephalae,fortune euonymus,radix sanguisorbae,herba lycopi,hedyotis diffusa willd,pteris multifida poir,costusroot,radix glycyrrhizae preparata,as 11 herbs in this formula,were identified by TLC（Thin layer chromatography）method;The gallic acid content was measured by HPLC method.（4）Stability study:Three batches of products were prepared by the optimal preparation process.Stress test,accelerated stability test and long-term stability test were carried out on these three batches of products.Results:（1）Fingerprint study based on standard decoction:26 common peaks were marked in the HPLC fingerprints of the standard decoction,which can be identified in the corresponding positions in the fingerprints of water extract,intermediate and finished product.The similarity of the fingerprints of water extract,intermediate and finished product to the one of standard decoction was respectively 0.969、0.958、0.968.The content of gallic acid in the formula’s ingredients was 100%.The tranfer rate of gallic acid was82.06%in standard decoction,90.75%in water extract,76.30%in intermediate,and 71.8%in the final product.18 common peaks out of the 26were attributed to the formula’s ingredients.（2）The preparation process of Jianpi Huoxue Jiedu granules:（1）Single-factor experiment:The crush degree of the ingredients was coarse.Soaking was not required.12 times the water was added.Decoction was two times and 1 hour each.Orthogonal experiment:16 times the water was added to the ingredients.Decoction was three times and 1 hour each.About 0.5m L of Volatile oil was collected to keep in reserve.（2）Forming process:Dextrin and soluble starch were added at 2:1 as excipients to mix with the clear paste,and screen with sieve of 16 mesh,dry at 60℃and constant pressure for 1.5 hours,spray with volatile oil.（3）Quality standard:The quality of the finished product met the standard required by general granules section in Chinese Pharmacopoeia 2015 edition;The TLC was established for Rhizoma atractylodis macrocephalae,fortune euonymus,radix sanguisorbae,herba lycopi,hedyotis diffusa willd,pteris multifida poir,costusroot,radix glycyrrhizae preparata.The TLC of the investigated ingredients showed clear spots,was specific and non disruptive to negative samples.The reference standard for the granules was set to be the content of gallic acid（C₇H₆O₅）in the radix sanguisorbae,which is at minimum 1.75mg per 1g.（4）Stability study:The sample showed serious water absorption in high humidity.The critical relative humidity of this product was measured to be 68.1%.At high temperature,the TLC spots of Rhizoma atractylodis macrocephalae and costusroot blurred.Under strong light all checks have passed.Products in blister packs have passed all the checks in accelerated stability test and 6-month long-term stability test.Conclusion:In comparison with the standard decoction,this product has small material changes during extracting,forming and drying process,which ensured the consistency of the final product and the former formula’s ingredients.The screened optimal preparation process was simple and feasible.The quality was under control and the products have high stability.The quality standard can control the granulation quality.This product should use blister packaging and be kept in a cool place with maximum 68%relative humidity.