Research Of Ornithine Aspartate Injection

Ornithine aspartate injection is widely used for hepatic encephalopathy treatment. The aims of this research are:(1) to establish a method for pilot scale experiment of ornithine aspartate injection preparation;(2) to development a method for the evaluation of the trial product and standard for its quality control of ornithine aspartate injection;(3) to study the stability of the trial product.The whole work included three parts:the optimization of the formulation.(1)the pH ranges of the ornithine aspartate injection were studied first and pH 5.7~6.7 were chosen as the suitable pH range;(2)the screening of the species and dosage of the antioxidants in prescription were been carry out. Our study found that the content of most of the impurities can be reduced through adding antioxidants, however, the content of impurity A become higher along with the amount of antioxidants increasing. So the method of fill nitrogen protection was adopted to control the increase of the impurities;(3)the dissolution temperature of pharmaceutical ingredients, the ways of fill nitrogen protection, and the amount of activated carbon and its adsorption time has carried on the exploration,(4)the best process conditions and their parameters were determined;(5) the packaging material was investigated, and brown glass ampoule bottle as the best was employed.Based on the process features of the product, the new standard is more rigorous, scientific. High Performance Liquid Chromatography(HPLC)methods were established to determine the content of the related substance. Themobile phase consisted of acetonitrile/phosphate buffer(Potassium dihydrogen phosphate 13.61 g, water 500 ml dissolved after add 25%- 28% of the ammonia water 5 ml, then add water diluted to 1000 ml, after blending, with phosphoric acid adjust pH value to 4.2 ± 0.1)(60:40).The detective wavelength was set at 205 nm. Based on the above method, the possible five kinds of impurity were effective detection and separation.The content is determined by high performance liquid chromatography too, The chromatographic conditions of content determination was same with that ofrelevant material, the methodology validation, including of recovery rate, precision, limit of quantitation, were carried out and their indexes meet the requirements.Amino acid ratio determined also by high performance liquid chromatography, the chromatographic conditions of the determination of amino acid ratio was same with that of relevant material. However, the original method was greatly improved toavoiding the instrument system error, the results of the methodology validation showed the validation indexes accord with the requests.Endotoxin determined by the limulus reagent method in detailed research, the result showed that the amount of endotoxin in each 1 ml contains should be less than 10 EU, and the amount can replace the imported standard execution in rabbit pyrogen method test, the method can save time cost and improve the economic benefit.The stability studies were carried out including influence factors test, freeze-thaw test, Accelerated test and long-term test. The results showed that the qualityof ornithine aspartate injection made by ourselveshave no obvious differencewith imported sample in characters and the ratio and content of amino acid.The impurity content of ornithine lactam made by ourselves significantly lower than that of the imported samples. The impurity content of ornithine lactam made by ourselvesdoes not exceed 0.8% after accelerating at 6 months and long-term investigation12 months, butthe impurity content of import ornithine lactamsamples has reached 2.85%in the same period.compared with imported samples, the overall quality of ornithine aspartate injection made by ourselves obtained improved obviously. The impurity control of ornithine lactam also had significantly improved. Moreover, the content of impurities in samples made by ourselves was far lower than the import samples, andindustrial mass productioncan realized through pilot production validation.