Improved validated assay for the determination of mefloquine and its carboxy metabolite in plasma, serum and whole blood using solid-phase extraction and high-performance liquid chromatography

An improved high-performance liquid chromatography method using a low silanol activity octadecylsilica column and a solid-phase extraction technique is validated for the simultaneous analysis of mefloquine and its carboxy metabolite in whole blood, plasma and serum. An octadecylsilica column with high silanol activity is compared to a column of low activity in terms of pH dependent variability of chromatographic retention times for mefloquine and its carboxy metabolite. The low silanol activity column showed a relatively large mobile phase pH range where retention times for both components are consistent. The solid-phase extraction procedure consists of a simple protein precipitation step followed by sample concentration and extraction using a C-18 membrane disk. The inter- and intra-assay variability for a therapeutic concentration of mefloquine (1000 ng/ml) is less than 2% in whole blood, plasma and serum while carboxymefloquine (1000 ng/ml) is 2.3% or less. At concentrations as low as 100 ng/ml the inter-assay variability is 6.2% or less for both analytes. This method shows a robust analytical procedure for the simultaneous analysis of mefloquine and its carboxy metabolite where precise measurements are useful in pharmacokinetic studies and in estimating drug compliance. (C) 1999 Elsevier Science B.V. All rights reserved.