| Bacterial Endotoxin Test(BET) are widely used to detect endotoxin in pharmaceutical industry. Based on the clotting reaction of Limulus Amoebocyte Lysate Reagent with endotoxin, BET is used for judging whether endotoxin limit comply with requirement. As one of the primary test items of drug quality control, BET plays an very important role to safeguard the usage of drug in the clinic application.Many factors influence the standardization of BET, one of the most important factor is Reference Standard Endotoxin. The import and establishment of RSE promote the BET standardization in great degree, harmonize and unify it in the range of the world.Despite many countries' RSE are prepared by hot phenol-water from E.coli, the certain difference of chemical structure, physical character and biological activity etc aspects exists in endotoxins by different extract methods. The material of national standard endotoxin(NSE) used in our country was produced by Shanghai institute of biological product in 1997, although it satisfied the request as standard material on biological activity and its biological character was analysis partly in the beginning. It has no any physical and chemical data, so it cannot compare with international standard and other countries' RSE, at the same time, it has been used nearly eight years, the stock are gradually dropping off, so we have to prepare new NSE as soon as possible.The main purpose of this experiment is studying the extract method, the assay method of physical, chemical character and biological activity of NSE's material to hope prepare new NSE soon.First, we established the extract method of NSE's material. According to the related literature and information, we selected four methods to extract endotoxin from E.coli O111:B4. Comparing endotoxins' biological activity and impurity content extracted by different methods, the results showed that the endtotoxin by Japanese method has stronger biological activity and lower impurity content. So we selected... |
PDF Full Text Request