Liquid chromatographic analysis of mesna and dimesna in plasma and urine of patients treated with mesna

Abstract

We describe in this report an expedient and accurate liquid chromatographic method for measurement of mesna and dimesna in plasma and urine. The separation of mesna and the internal standard (p-aminobenzoic acid, IS) was achieved on a 10-microns, 8 mm (i.d.) x 10-cm C18-Resolve cartridge in conjunction with radial compression system. An aqueous solution of sodium citrate (0.1 M), tetrabutyl ammonium phosphate (0.001 M), and triethylamine (1:10,000, vol/vol), adjusted to pH 5 with 85% phosphoric acid was used at a flow rate of 2 ml/min as a mobile phase. The compounds were detected in the effluent electrochemically at +450 mV. After an appropriate amount of IS was added, the plasma sample (100 microliters or fraction thereof) was deproteinized with an equal volume of 0.0825 M sulfuric acid containing sodium hexametaphosphate (1.25% wt/vol), whereas urine was diluted 1:50 with water and mixed 1:1 with an aqueous solution of sodium hexametaphosphate (1.25% wt/vol). Dimesna was reduced back to mesna with sodium borohydride before analysis of the total mesna. The peak height ratio (drug/IS) varied linearly with the concentration, and the correlation coefficient was > 0.992 for both mesna and dimesna. The intrarun precision at different concentrations of mesna was equally good and the coefficient of variation was consistently < 4.5%. No interference from endogenous substances or any concomitantly used drug was observed. This assay is currently being used for measurement of mesna and dimesna in plasma of bone marrow recipients who receive high doses of cyclophosphamide.