| Qihong Maitong injection (QMI) that composed of Astragalus and safflower herbs is a freeze-dried powder injection,which is clinical for treatment of Energy deficiency and Blood stasis caused by angina pectoris.It is still in the clinical research stage.The original preparation of the injection process is relatively mature,but the Passive allergy test of the finished security problems appears weakly positive occasionally.Based on the original preparation process and product validity,we tried to optimize the pretreatment process of this product by establishing a better evaluation system.The most important evaluation technology in this system was the membrane separation technology (the ultrafiltration technology),whose adaptability was systematically investigated in this study. All these optimizations could improve the safety and quality of Qihong Maitong injection,and provide a solid experimental evidence for membrane separation technology in the industrial application of the traditional Chinese injections as well. The main research contents and results are as follows:1. Preliminary establishment of evaluation system of the membrane separation processThe preliminary evaluation system was established as the following three aspects:(1) Index components determination:Establish methods to detect the components of hydroxy safflower yellow pigment A (HSYA),total saponins of astragalus and astragaloside.The methodological study showed that the methods were highly accurate,stable and viable.(2) Protein detection: Based on the examination of related substances in traditional Chinese medicine injections,we determined the quantity of protein by using the Bradford method. Results showed that this method was easy to operate with high sensitivity and less interference. Importantly,this method could enhance the efficiency of impurity removal during the subsequent pretreatment and ultrafiltration processes.(3) Fingerprint determination:In order to control the ingredients before and after ultrafiltration and the quality stability of the product, we did preliminary studies on the fingerprint detection of Qihong Maitong injection. All these detection methods above provided a more comprehensive evaluation system for further study.It also established a strong basis for the improvement of the quality and safety of the injection.2. Studies on the pretreatment process of Qihong Maitong InjectionThe optimization of heat treatment process of intermediates with liquid:Through the single factor test,we determined the optimal dissolution time for5min, and the heating temperature at80℃.Results showed that the high-temperature(≧80℃)or prolonged heating caused significant changes to the index components.The pretreatment process:We adopted refrigeration, centrifugation,and activated carbon adsorption for removal of impurities.Through the single factor test and orthogonal experiment we changed various process conditions,and the optimized process parameters are as follows:refrigerated for24h,centrifugated for15min,10000r/min,0.3%actived carbon adsorbed at40℃for30min,the original pH value.This process was simple to operate and the index components had a less loss,which could effectively remove the solid impurities, proteins,and resin.3. Adaptability research of membranes separation technique of Qihong Maitong InjectionPolysulfone, polyether sulfone, polypropylene and blended composite hollow fiber ultrafiltration membrane with the same entrapment of relative molecular mass of100000for ultrafiltration were selected to determine the best appropriate ultrafiltration membrane material. The results showed that the suitability of these four materials differs from each other.Briefly, both of the polysulfone and blend compound materials had a lower component permeation rate and a higher loss of active ingredients. What’s worse,polysulfone had a smaller membrane flux, longer ultrafiltration time and poorer pure water flux recovery rate and could easily lead to membrane fouling.Therefore,these two kinds of membrane material were not suitable in our solution system.However,polypropylene and polyether sulfone materials were much better. What’s more,polyether sulfone material had a lower removal rate of solid components,proteins and bacterial endotoxins than polypropylene material. The ultrafiltration results of Polypropylene hollow fiber ultrafiltration membrane is as follows:membrane water flux recovery rate is97.8%, index components transmittance of HSYA A, total saponins of astragalus and astragaloside are91.47%,91.44%,99.29%, bacterial endotoxin removal efficiency of100%.Taken together,the polypropylene-100000ultrafiltration membrane was suitable for the purification of Qihong Maitong injection.The polypropylene hollow fiber ultrafiltration membrane and the polyether sulfone plate ultrafiltration membrane with the same entrapment of relative molecular mass of100000and50000were selected to determine the best appropriate ultrafiltration membrane.Results showed that neither of the membranes with the entrapment of relative molecular mass of50000fit the drug solution system due to the great loss during ultrafiltration;The membranes with the entrapment of relative molecular mass of100000had the suitable permeation rate for index component,and could effectively remove pyrogens.However,the resin removal effect of hollow fiber ultrafiltration membrane was poor.The ultrafiltration results of the polyethersulfone plate ultrafiltration membrane with the entrapment of relative molecular mass of100000is as follows: index components transmittance of HSYA A, total saponins of astragalus and astragaloside are85.72%,81.52%,82.33%, solids removal rate of17.99%, protein removal rate of21.22%. The resin checks and the pyrogen test checks are in line with index requirements of the Chinese medicine injections.In short,the polyethersulfone plate ultrafiltration membrane with the entrapment of relative molecular mass of100000is applied to purification of the drug solution system.After that we tended to ascertain the appropriate ultrafiltration membrane for the II ultrafiltration process from the four kinds of membranes mentioned above.Compared with the I ultrafiltration, the solid and protein reduction rate of the two kinds of50000membranes was higher during the Ⅱ ultrafiltration,but the index components transmittance was lower.Therefore, the II ultrafiltration process is not applied to purification of the drug solution system.4. Studies on optimizing of the membrane separation processAccording to the results of adaptability study,we tried to optimize the operating conditions of the plate ultrafiltration membrane with the entrapment of relative molecular mass of100000. Through the single factor test, the liquid temperature and operating pressure of ultrafiltration were optimized respectively.The optimized conditions are as follows:liquid temperature30℃, operating pressure O.1Mpa.These conditions led to a high efficiency of ultrafiltration and a less loss of index components.5. Studies on the safety and major pharmacodynamics of Qihong Maitong InjectionWe next evaluated the safety of the finished products of Qihong Maitong injection.Results of muscle and blood vessel irritation test,hemolysis test and active allergy test were all negative. The result of passive allergy test showed that the finished products of the original preparation process was weakly positive,while the products of the optimized process was negative,which indicated that the application of ultrafiltration technology could effectively eliminate passive allergic reactions and improve the safety of the injection. A comparative study of the major pharmacodynamic between the original process finished products and the optimized process finished products showed that the pharmacodynamic of the two crafts is equal. |
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