| ObjectivesTo make an all-around and systematic study of process optimization of Qingkailing Injection,and give explicit technological parameters of production process which are thought to be related to preparation safety to ensure product quality.MethodsThe research optimized the technological parameters of production process,including the extraction,hydrolysis,purification of herbs and matching liquid to determine the best production processes.It provided a effective support for the good stability and uniformity of Qingkailing Injection.1.To optimize the extraction process of radix isatidis,gardenia and honeysuckle.(1)Radix isatidis,gardenia and honeysuckle need to be extracted in the production process of Qingkailing Injection.By the content of epigoitrin,geniposide,chlorogenic acid and galuteolin respectively,the research explored the water amount to optimize the extraction of radix isatidis,gardenia and honeysuckle.(2)To provide objective foundations for concentration,the relative density of concentrated liquid in the production process of Qingkailing injection was first detected.2.To optimize the purification process of radix isatidis and honeysuckle.(1)Taking the content of epigoitrin as an evaluation index,the study optimized the purification process of radix isatidis,including ammonia precipitation and decolorization with activated carbon.(2)Three influencing factors in the alcohol precipitation techniques,including the fluid temperature,the stirring speed and the speed of adding alcohol,were optimized by the orthogonal test.The content of chlorogenic acid and galuteolin were taken as comprehensive indicators,and multi-index comprehensive scoring method was used in the data analysis.3.To optimize the purification and hydrolysis process of bubali cornu and concha margaritifera.(1)Inductively coupled plasma mass spectrometry(ICP·MS)was first introduced to make the maximum acceptable concentration of barium ions of the hydrolysate.The orthogonal test was adopted to optimize the hydrolysis process by the content of barium ions and nitrogen.(2)The relative density and the influence of pH hydrolysis have been studied.In addition,the molecular weight distribution of protein in hydrolysate was determined using SDS-PAGE.4.To optimize the confecting technique of Qingkailing injection.(1)Taking the comprehensive score of active components as an indicator,the subject optimize the confecting technique,membrane pore size of ultrafiltration technology and sterilization with activated carbon.(2)With the investigation of the method of nitrogen adding,the paper determined the appropriate filling style to ensure the stability of Qingkailing injection.(3)Based on the process optimization of intermediates,the paper investigated the storage time.Results1.The optimization of the extraction process of Qingkailing injection.(1)The result of the water addition suggested that the amount of radix isatidis and gardenia extraction process for the first time was 6-fold of the material,and the second time was 4 and 5 time respectively.The water amount of honeysuckle extraction was 10 times for the first time and 6 times for the second.(2)The specific results of concentrated liquor of radix isatidis,gardenia and honeysuckle were as follows:(1.1167±0.0251)g·mL-1(60?)?(1.0833±0.0153)g·mL-1(600?)?(1.0933±0.0153)g·mL-1(60?).2.The optimization of the purification process of radix isatidis and honeysuckle.(1)The study of the ammonia precipitation showed that adjusting pH to 8.50 with ammonia and adjusting pH to 6.10 after removal of ammonia by heating could improve the content of effective component.The optimal eonditions of decolorization were 1%of activated carbon,and bleaching time 15 min.(2)The best alcohol-precipitation procedure was determined as adding alcohol with the speed of one time of the material per minute at 20?,and the stirring speed was 240 rpm.3.The optimization of the purification and hydrolysis process of bubali cornu and concha margaritifera.(1)The optimal hydrolysis conditions were obtained:bubali cornu was 1.5 mol·L-1 Ba(OH)2 for 7h and concha margaritifera was 1.5 mol·L-1 H2SO4 for 7h,the amount of Ba(OH)2 and H2SO4 is 8 times of the material.The content of barium ions of the hydrolysate should less than 0.2?g/g.(2)After combining hydrolysate,the mixture should be adjusted pH to 3.80,its relative density was(1.0163±0.0020)g·mL-1(60?).The hydrolysate molecular weight is range from 4.6 to 10 KDa.4.The optimization of the confecting technique of Qingkailing injection.(1)It turned out that the proper mixing sequence is to add hydrolysate,radix isatidis extracts and gardenia extracts in order,and then add them to 75%ethanol solution of cholalic acid and hyodeoxycholic acid to make mixed liquid of six.The pH value of the ethanol precipitation was 7.0 and the injection preparation was 7.50.The result indicated that activated carbon dosage was 0.5%and the pore size of ultra filtration membrane was 30KD.(2)The paper determined the appropriate nitrogen filling style was fill twice,Pre and post,eould keep the effective components of preparation stable.(3)The survey of storage time showed that extracts,hydrolysis and intermediates were stable within 72 hours.ConclusionThe paper established a quantitative method for Qingkailing injection intermediate and product through different analysis and testing technologies·The production process was optimized based on the process and information provided by pharmaceutical factory.Since the level of parameters control for alcohol-precipitation is quiet low and the hydrolysis process of bubali cornu and concha margaritifera could introduce large molecular substances,which will cause safety risks.The subject adopted the multi-target orthogonal experiment for an in-depth study of the alcohol-precipitation and hydrolysis process.In addition,the relative density of concentrated liquid in the production process of Qingkailing injection was first detected to provide objective foundations for concentration.The above researches provided a effective support for the good stability and uniformity of Qingkailing Injection.It eliminated the bottleneck of differences among product batches,which was caused by the low level of parameters control.Thus the adverse reactions of Qingkailing injection are reduced at the source.The paper has great value to keep stable in quality and in ensuring clinical safety and effectiveness. |
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