After 30 min at room temperature, the reactions were stopped by the addition of 40 μl of sodium hydroxide solution (at a concentration of 0.1, 0.4, 0.8, 1.0, 1.5, 2.0, or 3.0 M, respectively). Louis, Mo.) in rat plasma was determined by mixing 150-μl samples of drug-free rat plasma, freshly spiked with CMS (25 mg/liter of plasma), with 30 μl of netilmicin sulfate solution (5.0 mg/liter, internal standard Schering-Plough, Kenilworth, N.J.) and 20 μl of sulfuric acid solution (0.1, 0.4, 0.8, 1.0, 1.5, 2.0, or 3.0 M). It involves accelerated hydrolysis of CMS to colistin in biological fluids, extraction of the generated colistin, and derivatization prior to HPLC analysis according to a previously described method for the quantification of colistin.The optimal concentration of sulfuric acid for the hydrolysis of CMS (Sigma, St. The present report describes a simple and sensitive HPLC method for the assay of CMS in plasma and urine. Microbiological assay is currently the common analytical method for measuring CMS in biological fluids.
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