Study On The Quality Standard Of Freeze-Dried Hirudo Nipponica And The Hirudo Nipponica Seedlings

Objective: Freeze-dried Hirudo nipponica is a new type of processed product obtained through low temperature drying.To ensure its quality and safety,this study eventually developed a quality standardby examining its stability,safety,toxicology and pharmacodynamics.This study developed quality grading standards and seedling quality standards by studying its egg cocoon and seedlings,it has profound significance for the long-term development of Chinese herbal medicines leeches.Method: 1)A series of suitability experiments were conducted to establish the microbiological detection method for freeze-dried Hirudo nipponica.2)The effects of temperature,humidity and strong light irradiation on the freeze-dried Hirudo nipponica were investigated,as well as accelerated tests to determine the packaging and storage conditions.3)Through the maximum tolerance test(MTD),the safety of the dosage was investigated.The effects of BT andCT on mice and acute blood stasis in rats were used to examine efficacy.4)Character and TLC are used for its identification.The water,total ash,acid insoluble ash content,PH,heavy metals and harmful elements,aflatoxin,microbial limit to control and antithrombin activity test are used for controling the quality of freeze-dried Hirudo nipponica,and establishing quality standards.5)The quality grading standards.of egg cocoon was established by studying the weight,width,seedling rate and incubation rate of the cocoon.6)Through the study of the morphology,weight,length and vigor of Hirudo nipponica seedlings,the quality standard of seedling was drafted.Results: 1)Freeze-dried Hirudo nipponica is basic no bacteriostasis vermiculite,vaccination five positive experimental strain recovery ratio isbetween 0.5 ~ 2.0,therefore can adopt the method of plating(routine method,1 ml)per dish for microbial count.2)Under the conditions of high temperature,high humidity and strong light,the activity of antithrombin was obviously decreased.In the accelerated test,the antithrombin activity of the freeze-dried Hirudo nipponica was slowly decreasing,and the activity of antithrombin in the 6th month was still higher than that of 200U/g.3)The maximum tolerance test dose was 26.4g /kg,which was 58 times the maximum dosage of the pharmacopoeia,and no death or adverse reactions occurred in the mice.It can obviously prolong the BT and CT of mice,and prolong the PT,TT and APTT of acute blood stasis syndrome,and have strong anticoagulant effect.It can inhibit platelet aggregation in rats and reduce the whole blood viscosity of rats.However,there was no obvious effect on Fibg and plasma viscosity.4)In TLC chromatogram of the test,the same magenta spot was shown in the corresponding position of the control drug chromatogram,and the same orange red fluorescence spot was shown under the ultraviolet light(365nm).Increased microbial detection,and the contents,water,total ash,acid insoluble ash,PH and aflatoxin were all higher than those in the pharmacopoeia.5)Selecting weight as the basis for the classification of freeze-dried Hirudo nipponica egg cocoon,the weight of the first egg cocoon was 0.08 g ~ 0.16 g,and the incubation rate was 90%,and the seedling rate was6.78.6)There were differences in body length,weight and vigor of seedlings in different ages,and the solid line was growth slower than the dotted line.Conclusion: Hirudo nipponica have high antithrombin activity,but traditional processing methods such as traditional drying and ironing can destroy the active ingredient hirudin.In addition,the leech medicinal materials which is susceptible to Aspergillus flavus and other microorganisms without packing and placed at random.Compared with traditional processed products,the antithrombin activity of Hirudo nipponicawas greatly improved.Therefore,establishing the quality standards of freeze-dried Hirudo nipponica to ensure the authenticity and anticoagulant activity.meanwhile,in the condition of refrigeration,it can eliminate the possibility of infecting aflatoxin and other microorganisms at the circulation and storage link which use low-density medicinal polyethylene bag as direct contact inner packing of the medicinal material and use pet/al/PE composite membrane as outer packing material.It has improved the biological potency of leech and improved the safety of medicinal materials.In this study,the weight,Seedling emergence rate and incubation rate of Hirudo nipponica egg cocoon were studied,and the quality grading standard of egg cocoon was established.By studying the morphology,length,weight and vitality of Hirudo nipponica seedlings,the quality standard of Hirudo nipponica seedlings was drafted.Hirudo nipponica eggs cocoon grading standards can ensure the quality of eggs cocoon.Hirudo nipponica seedlings quality standards can ensure the quality of seedlings and provide reliable seedlings,it has a great significance to develop the artificial cultivation of Hirudo nipponica and relieve the shortage of the market resources of leech medicinal materials.