Topic 1: Research On The Preparation Process And Quality Standard Of Jiedutongluo Drop Pills Topic 2: Research On The Preparation Process And Quality Standard Of Ophiopogon Japonicus Total Saponin Drop Pill

Purpose and significanceAccording to the epidemiological investigation and analysis,stroke disease has been becoming the diseases with the highest incidence rate in China in recent years,and its disability rate is also ranked first in the world;Chinese medicine believes that cerebral stroke is related to poor blood flow in the brain and congestion.Therefore,activating the circulation,promoting blood circulation and removing blood stasis have become the main methods for treating brain stroke in traditional Chinese medicine.Jiedu Tongluo precription is the academic experience of Wang yongyan academician.The precription is mainly composed of three drugs:total saponins,gardenia extract and scutellarin.In the prescription,the gardenia is a king for detoxification;Gardenia extract is the main drug used to detoxify,total saponins and scutellarin are auxiliary drugs used to Remove stasis,three drugs are organic compatibility to cut the pathogenesis of stroke.Due to the limitation of traditional decoction using,the quality of the production process is difficult to control stably.Therefore,this group is guided by the theory of traditional Chinese medicine,combined with modern Chinese and medicine technology,and it is planned to develop this prescription into a proprietary Chinese medicine according to the relevant requirements for registration of new drugs.It had provided a good scientific basis for the development and application of traditional Chinese medicine.Methods:1.preparation technology research of dropping pillsThe research group preliminarily established the preparation process for understanding Tongluo Dripping Pills.Due to the phenomenon of inhomogeneous liquid mixing,it was found through studies that the problem of unevenness was mainly caused by the inconsistent mixing of gardenia and breviscapine extraction.To solve this problem,we optimized the preparation process of Jiedu Tonglu Dripping Pills.The single factor test method was used to determine whether the roundness of the dropping pills,the uniformity of mixing of the liquid and the base,and the weight difference were used as indicators,investigating the types of substrates required for the preparation of Jiedu Tongluo Driping Pills,the method and sequence of drug addition,the proportion of drug and matrix,the type of condensing agent,the dripping temperature,droplet distance,drip rate,cooling column length,and cooling temperature.2.Study on the determination of active componentsThrough literature and research,It was found that the seven active ingredients in Jiedu Tongluo Dropping Pills can be completely dissolved after sonication in methanol,Therefore,in this project,the sample of Jiedu Tongluo Pills was prepared by methanol ultrasonic treatment.The ultrasonic treatment time of Jiedu Tongluo Drop Pills was:30 min for gardenia extract and total saponins,and 60 min for scutellarin.The ultrasonic power and frequency are 300 W and 50 KHz,respectively.Using high-performance liquid chromatography("2015 edition of the Chinese Pharmacopoeia"fourth part of the preparation of general rules 0512),established a method to determine the content of gardenia extract,total saponins,and scutellarin in the Jiedu Tongluo Droping Pills.3.Dissolution studies of dropping pillsAccording to the small cup method in the dissolution and release assay of the fourth edition of the 2015 edition of the Chinese Pharmacopoeia,a volume of dissolution medium of 250 mL,and a temperature of 37℃,speed 50 r/min,the time groups were sampled at 3,6,10,15,20 and 30 minutes,sampling 1.5 mL,immediately after sampling,adding 1.5 mL of isothermal medium.The eluate was removed by filtration using a 0.45 μm microporous filter and 20 μL of the solution was accurately pipetted into the HPLC.Six samples were prepared in parallel according to the method described above.(Dissolution medium:water was selected as the dissolution medium for gardenia extract.Total saponins,and scutellarin were pH 6.8 phosphate buffered saline as the dissolution medium).4.Quality standards researchIn this part,Quality Standard of Jiedu Tongluo Droping Pills was study initially,which according to the request of the granules in 2015《Pharmacopoeia of the people’s republic of China》.The method to test the content of loganin and morroniside were established.With a view to provide the basis for the future quality control of Jiedu Tongluo Dropping pills pilot test products.Results:1.Preparation technology optimization research of dropping pillsTake three kinds of drugs,respectively powder,screen 120 mesh sieve.The PEG1500 was melted in a boiling water bath,and wild astaxanthin was added and stirred uniformly.After the scutellarin was completely dissolved,PEG4000 was added and stirred well;Weigh the wolfberry fruit extract and add 40%water to stir to obtain the wolfberry fruit extract liquid;slowly add the scutellarin glycoside liquid to the gardenia extract liquid,stir it until it is even,add the total stir the saponin uniformly,keep it warm for 30 minutes,pour it into the dropping pill machine,drop it into the liquid paraffin and take it out.Form it with a filter paper to remove the condensing agent.(The ratio of drug to matrix is 1:3.6,PEG4000:PEG1500=1:3.6,dropping temperature is 80 ℃,condensing agent is liquid paraffin,condensing agent temperature is 0～4 ℃,cooling cylinder length is 50 cm,drop distance is 2～4cm,Drop speed 60 drops/min.)2.Study on the determination of active componentsSample preparation:take 20 drops of dripping pill sample and finely weigh dripping powder in a volumetric flask,add methanol ultrasonic,put it to room temperature,add methanol to constant volume,shake well and get it.Through the experiment concluded:the three established methods of specificity,linearity,precision,repeatability,and rate of recovery meet the content determination requirements and they can be used as a method for determining the content of active ingredients.Three batches of Detox Tongluo Pills were prepared according to the optimized dropping pill forming process.The average content of geniposide in the three batches of Jiedu Tongluo Drop Pills was determined by high performance liquid chromatography to be 15.8 mg/g.The content was 13.66 mg/g,the average content of notoginsenoside R1 was 11.10 mg/g,the average content of ginsenoside Rg1 was 35.16 mg/g,ginsenoside Re was 3.8 mg/g,ginsenoside Rbl was 37.89 mg/g,and ginsenosides Rd is 10.98 mg/g3.Dissolution studies of dropping pillsThe vitro dissolution rate of Jiedu Tongluo dripping pills determined by high performance liquid chromatography showed that the seven active ingredients in Jiedu Tongluo dripping tablets dissolved completely in 15 minutes,and the cumulative dissolution percentage was in the range of 97%～103%,so Jiedu Tongluo dissolution of the seven active ingredients in the dripping pills meets the requirements and reflects the instant characteristics of the dripping pills.4.Quality standards researchEstablish a method for determining the contents of geniposide,scutellarin,and total saponins in Jiedu Tongluo Drop Pills.Geniposide:Take 20 drops of dripping pills,sifting through 60 mesh,take 0.15g of powder,accurately weigh,place in 25 mL volumetric flask,measure 20 mL of methanol as solvent,ultrasonic for 30 min(power 300 W,frequency 50 KHz),room temperature,add methanol to the mark,mix well,and sample the 0.45 μm filter to obtain the test solution.The content was measured on a Kromasil-C 18 column(4.6 mm×150 mm,5 μm,column number 1815045);acetonitrile-water(15:85)as the mobile phase;the detection wavelength was 238 nm;the injection volume was 10 μL;1.0 mL/min;column temperature 30℃.Scutellarin:Take 20 drops of dripping pills,sifting through 60 mesh,take 0.11g of powder,accurately weigh,place in 25 mL volumetric flask,measure 20 mL of methanol as solvent,ultrasonic for 60 min(power 300 W,frequency 50 KHz),remove and put it to room temperature,add methanol to the mark,mix well,and sample the 0.45 μm filter membrane to obtain the test solution.The content was measured on a Kromasil-C 18 column(4.6 mm× 150 mm,5 μm,column number 1815045);methanol-0.1%phosphoric acid(40:60)was used as the mobile phase;the detection wavelength was 335 nm;the injection volume was 5 μL;The flow rate was 1.0 mL/min;column temperature 40℃.total saponins:Take 20 drops of dripping pills,sifting through 60 mesh,take 0.15g of powder,accurately weigh,place in 25 mL volumetric flask,measure 20 mL of methanol as solvent,ultrasonic for 30 min(power 300 W,frequency 50 KHz),room temperature,add methanol to the mark,mix well,and sample the 0.45 filter to obtain the test solution.The determination was performed on a CAPCELL PAK C18 column(4.6 mm × 250 mm,5 μm,column number AKAD 12731);acetonitrile was used as the mobile phase A and water was used as the mobile phase B(elution conditions:0 to 30 min 19.6%B.,30～60min 19.6%→39%B,60～63min 39%B,63～70min 39%～60%B)Column temperature 25℃;Detection wavelength:203 nm;Injection volume 10 μL;Flow rate:1.5 mL/min.Conclusions:This topic has optimized the molding process of Jiedu Tongluo Dripping Pills and established a method for determining the content of the components in the Tongluo Dripping Pills based on the 2015 edition of the Chinese Pharmacopoeia.The purpose of this study was to investigate the in vitro dissolution of Dudu Tongluo Pills.Initially establish the quality standards for understanding Tongluo Dripping Pills,and provide reference for the quality control of Jiedu Tongluo Dripping Pills in the later period.Purpose and significanceA tumor is a new organism formed by the proliferation of local tissue cells under the action of various tumorigenic factors..In most patients with tumors,tumor metastasis is the cause of death in 90%of cancer patients,so inhibition of tumor metastasis is one of the effective strategies for tumor treatment.In recent years,some scholars have found that the total saponins of ophiopogon japonicus have a good effect of inhibiting tumor metastasis,and have the advantages of low dose,high safety,and no cytotoxicity to normal cells.Pills have the advantages of simple preparation process,improved bioavailability of drugs,rapid efficacy,and small drug loading.Since the total saponins of Ophiopogon japonicus is hardly soluble in water and the daily dose is relatively small,it is proposed to use solid dispersion technology to prepare dripping pills by examining the dropping pills preparation process.This subject firstly studied the preparation process of Ophiopogon japonicus saponin dropping pills in the short-grass mountain,established a method for the determination of the total saponins of Ophiopogon japonicus,and initially determined the quality standards of Ophiopogon japonicus saponins dropping pills.The study provides good research ideas for the application and development of Ophiopogon japonicus saponins.1.Preparation process studyThe single round test method was used to determine the degree of roundness of the dripping pills,whether the liquid and the base were mixed uniformly,and the difference in weight was used as the index:the type of matrix,the order of adding the drugs,the ratio of the drug to the base,the heating temperature,the dripping temperature,the drop distance,Drop speed,cooling column length,cooling temperature.2.Study on the determination of active componentsThrough literature research,it was found that the two active ingredients in the Ophiopogon japonicus saponin dripping pill can be completely solubilized in methanol after sonication.Therefore,in this project,the total saponins of Ophiopogon japonicus were prepared by the method of methanol ultrasonication.Dropping pill sample.The time of sonication of the total saponins of Ophiopogon japonicus saponin was 30 min.The ultrasonic power and frequency are 300 W and 50 KHz,respectively.The short-grass-root ophiopogon japonicus-S and short-grass root ophiopogon japonicus-R are the target components in the total saponins of ophiopogon japonicus.The content of total saponins from Ophiopogon japonicus was determined by high performance liquid chromatography("2015 edition Of the Chinese Pharmacopoeia",Part 4 General Principles 0512),and the establishment of japonicus saponin-S and japonicus Ophiopogon japonicus-R was established.Determination of content.3.Quality standards researchIn this part,Quality Standard of Ophiopogon japonicus Droping Pills was study initially,which according to the request of the granules in 2015《Pharmacopoeia of the people’s republic of China》.The method to test the content of loganin and morroniside were established.With a view to provide the basis for the future quality control of Ophiopogon japonicus Dropping pills pilot test products.Results1.Preparation Process StudyTake the total saponins extract of Ophiopogon japonicus saponin,and screen 7 sieve;Melting PEG2000 in a water bath at 80℃.,and adding the total saponins of Ophiopogon japonicus Stir to mix thoroughly.After completely dissolve,add PEG4000 and mix evenly,heat for 30 minutes.Pour into a drop pill machine,drop it into liquid paraffin and remove it after forming.Use a filter paper to remove the condensing agent to obtain a dropping pill.(drug and matrix ratio 1:11～1:13,PEG4000:PEG2000=1:10,dropping temperature 80℃,condensing agent is liquid paraffin,condensing agent temperature 0～4℃,cooling column length is 50cm,drop distance 2～4cm,drop Speed 60drops/min.2.Study on the Determination of Active ComponentsSample preparation:take 20 drops of dripping pill sample and finely weigh dripping powder in a volumetric flask,add methanol ultrasonic,put it to room temperature,add methanol to constant volume,shake well and get it.Through the experiment concluded:the two established methods of specificity,linearity,precision,repeatability,and rate of recovery meet the content determination requirements and they can be used as a method for determining the content of active ingredients.Content determination results:the results of the content determination showed that:the content of Ophiopogon japonicus saponin-S was 15.37 mg/g;the content of Ophiopogon japonicus saponin-R was 23.04 mg/g.3.Quality standards researchEstablish a method for the determination of Ophiopogon japonicus Saponins-S and Spodoptera Spp.Ophiopogon japonicus saponins-R,which are the index components of Ophiopogon japonicus saponin dripping pills,by taking 20 drops of dripping pills.Mesh sieve,take powder 0.35g,accurately weighed,set in a 25 mL volumetric flask,measure 20 mL of methanol as a solvent,ultrasound for 30 min(power 300 W,frequency 50 KHz),remove to room temperature,add methanol to the mark,Mix evenly,sampled 0.45 filter,that is,the test solution.The content was measured on an XBridgeTM C18 column(4.6×250 mm,3.5μm,column number 186003943);the mobile phase was acetonitrile-methanol-ultrapure water(39:5:56);the detection wavelength was 203 nm;the injection volume was 20 μL;flow rate:1.0 mL/min;column temperature:40℃.ResultIn this study,the optimization of the molding process of Ophiopogon japonicus saponin dripping pills was studied.According to the 2015 edition of the Chinese Pharmacopoeia,the method for determining the content of the index components in Ophiopogon japonicus saponin dripping pills was established.The quality standard of Ophiopogon japonicus total saponin dropping pills provides a reference for the quality control of Ophiopogon japonicus saponins dropping pills in the later period.