| This project focuses on the implementation and validation of a reversed phase high performance liquid chromatography method for polymyxin B (PMX-B) in plasma. PMX-B is an antibiotic with a bactericidal action, used for the treatment of serious infections including infections of the urinary tract, meninges or blood stream caused by susceptible gram-negative bacteria (e.g. Pseudomonas aeruginosa, Escherichia coli, Enterobacter aerogenes, Klebsiella pneumonia, Haemophilus influenza). In the product information of PMX-B, there is no adequate information on pharmacokinetics and pharmacodymanics because methodical studies on Polymyxin pharmacokinetics were never conducted. Fundamental to PK/PD studies is the availability of a robust analytical method to quantify the drug in biological fluids.;Hence, the primary aim of the study is to establish a suitable analytical method for the quantification of PMX-B in human plasma. The analysis of PMX-B in biological fluids by conventional HPLC is difficult due to very weak UV absorption and no native fluorescence. In conclusion, this study is an attempt to establish, robust, and reliable analytical method for quantification. To achieve this goal we tried to implement and validate the HPLC method by formation of fluorescent derivative as described by G. Cao et al. We were able to successfully implement the method however the sensitivity was much lower than that reported in the reference 1000 ng/mL. Thus, several attempts to improve the quantification issue were done but unfortunately they were not effective. |
