Qing Ling Injection Of Macromolecular Protein Detection Research

Chinese Medicine Injection is an innovation of the modernization of traditional Chinese medicine, and it has played an important role in Medical Social Work. The practical applicability and effectiveness of Chinese Medicine Injection have brought huge economic results in society. But as the widely application of Chinese Medicine Injection, the reports of adverse reactions is increasing, too. The safety of Chinese Medicine Injection has attracted the attention of the society. The complexity of Chinese Medicine Injection’s composition, the difference of pharmaceutical technology, the lack of appraisal system, are all the factors of the security of Chinese Medicine Injection. It’s very important to strengthen the control of the Chinese Medicine Injection preparation technology and set up the evaluation system of Chinese Medicine Injection.This paper studied one of the most often occur adverse reaction Chinese Medicine Injections in recent yeas:Qingkailing Injection as the objects of study. We also select some Chinese Medicine Injections to prove the analysis method. These Chinese Medicine Injections included Astragalus Injection, Shengmai Injection, Yanhuning Injection. We aimed at the protein in Chinese Medicine Injections. We selected some analysis methods which often use in protein analysis and suitable for this study. Through the study of intermediates of Qingkailing Injection, we discussed the source of the protein in Qingkailing Injection, and this could provide reference to the process control of Qingkailing Injection.We used Coomassie birilliant blue and biuret colorimetry to determine the content of protein in Qingkailing Injection which had been precipitated by trichloroacetic acid. The objects of the study were Qingkailing Injection, Qingkailing Injection Intermediate. We also use Astragalus Injection, Astragalus Injection Intermediate, Shengmai Injection to prove the analysis method. Through the study, we got the approximate scope of the protein in the Qingkailing Injection. Qingkailing Injection took some reactions with Coomassie birilliant blue reagent, so we used biuret colorimetry to determine the content of protein which had been precipitated by trichloroacetic acid. The content of protein in Qingkailing Injection was0.1125mg/mL, The content of protein in Shengmai Injection Intermediate was0.416mg/mL We use Coomassie birilliant blue to test the others. The content of protein in Astragalus Injection was0.0496mg/mL. The content of protein in Astragalus Injection Intermediate was0.2336mg/mL. The content of protein in Shengmai Injection was0.017mg/mL The results showed that the content of Chinese Medicine Injection is higer than Chinese Medicine Injection Intermediate. The results provided a reference to loading quantity of SDS-polyacrylamide gel electrophoresis.We used SDS-polyacrylamide gel electrophoresis to investigate the molecular weight scope of the protein in Qingkailing Injection. We added several kinds of Qingkailing Injection Intermediates in this part.The results showed that there were high molecular weight protein in Qingkailing Injection Intermediates, and through the alcohol sink, the molecular weight decreaseed. Take Banlangen Concentrate and Banlangen alcohol sink solution as an example, after alcohol sink, the molecular weight was from3-80KD reduced to3-30KD. The molecular weight in Qingkailing Injection was3-17KD. We also use Astragalus Injection, Shengmai Injection to prove the analysis method. We detected no protein in Astragalus Injection, the biggest protein in its intermediate was58KD. The molecular weight in Shengmai Injection was17-175KD.We used SEC to for a further investigation of the molecular weight of protein in Qingkailing Injection and their intermediates. There was no peak in the chromatograms of Qingkailing Injection and Qingkailing Hydrolyzate solution intermediate. By calculating, the molecular weight of protein in Gardenia Hydrolysate was283KD. The molecular weight of protein in Honeysuckle was31KD. The molecular weight of protein in Banlangen Hydrolysate was42KD. We also use Astragalus Injection, Astragalus Injection Intermediate, Shengmai Injection to prove the analysis method. The molecular weight of protein in Astragalus Injection intermediate was64KD. The molecular weight of protein in Shengmai Injection was29KD. The molecular weight of protein in Yanhuning Injection was10KD. Although there was difference between SDS-polyacrylamide gel electrophoresis and SEC, the trend was consistent.We use MS to investe the structure of the protein in Qingkailing Injection and its Intermediate. We use the protein which was got from SDS-PAGE. The result shows:the messages which we got from four samples are similar. We guess there were the same protein in these samples. The studies above showed that SDS-polyacrylamide gel electrophoresis and SEC could use to analysis the protein in Qingkailing Injection, and to provide a renference to quality control of Qingkailing Injection.