- 學位論文
用高效能液相層析分析Colistin並探討市售產品含量差異
Analysis of Colistin by a Modified HPLC Method and Application to Analysis of Conditioning Colistin Products
摘要
Colistin化學名為Colistimethate sulfate（CMS），俗名Polymyxin E。1950年由日本小山田在Bacillus polymyxa var. colistinus菌所分離而得，屬於鎖環狀多胜體抗生素，亦屬抗生素polymyxin其中之ㄧ，但毒性比polymyxin B為低，且二者均屬於狹效性抗生素 ;對革蘭氏陰性菌具有抑制作用，尤以沙門氏桿菌，綠膿桿菌及大腸桿菌有很強的抑菌作用。 分析colistimethate sulfate主要以Fluorescence HPLC分析其水解產物polymyxin A, B之含量並再加入內標標準品來進行分析，但準確度不高、且操作不便。本實驗將樣品處理方法經過調整與改善，使colistimethate sulfate水解物與螢光物FMOC-Cl反應產生FMOC-(colistin)-NH2，再以FMOC-(colistin)-NH2為標的測定之；結果發現樣品經處理方式修改比文獻提供樣品經處理方式較容易進行HPLC分析，而本研究亦比較市售產品之國外原廠藥品則是將原料經過處理後再以凍晶方式貯存販售與國內學名藥將原料經過處理後再以粉充方式貯存販售。由於兩者來源不同且處理與貯存方式也不同，所造成CMS溶解後經水解含量，發現具有差異。並發現二者配製貯放在37 ℃時，放置約16小時發現其colistin含量最高，而放置約24小時後趨近平緩。最後進行微生物抑菌能力測試發現：存放在37 ℃時，放置約16小時發現其colistin含量最高，抑菌能力較大。然而以抑菌圈能力差異判讀並不會太過明顯；若以效價計算，國外原廠藥品較國內學名藥高。
關鍵字
高效能液相層析分析並列摘要
Colistin formal name is Colistimethate sulfate and popular name which is Polymyxin E. The Colistin was separated from Bacillus polymyxa var. colistinus by Japan Mr. Koyama in 1950s. It is belongs to a cyclic heptapeptide and polymyxins. However the toxicity is less more than polymyxin B and they are refer to constricted antibiotics. They could to inhibit the development of the Gram-negative Bacteria especially are Salminella, Pseudomonas aeruginosa, and Escherichia coli. The major detection of the method is to use the Fluorescence HPLC that to analyze the products of the colistimethate sulfate after dehydration. The products are contains polymyxin A and polymyxin B. Whatever this method must have the additive such as sample pretreatment included mixing standard solution. Exactly the precisely does not well and not easily to handle. The sample have to aseptic that to produce the dehydrated component and to reacte with fluorescent material like FMOC-Cl. To detect the products which is FMOC-(colistin)-NH2 and find the sensitivity and duplicate are the best more than the ordinary HPLC. Analysis to the brand name drug between the foreign manufacturer and the domestic laboratory could to discover the drugs had passed the dry products method that to sale and storage; to contrast the drugs about the internal generic drug that are to sale and storage by to stuff. We could notice the differentials the hydrated component after the CMS was dissolved by the drugs sources and different manners to storage and aseptic. Also to find the both was to storage at 37 ℃ after 16 hours and to detect the colistin that are still have strong concentration. But after 24 hours the concentration are beings to decrease gently. The experiment about to test the inhibition of the growth of the microorganisms: it is have the strong affect about the Colistin to inhibit the growth of the microorganisms that are to storage at 37 ℃ after 16 hours﹒However it is not significant to identify the value for inhibition circle zone and to calculate the potency that could to discover the brand name drug is higher than the generic drug.