HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC ASSAY FOR METHYL-BETA-CYCLODEXTRIN IN PLASMA AND CELL LYSATE

This paper describes a high-performance liquid chromatographic method with fluorescence detection for the analysis of methyl-beta-cyclodextr in (MEBCD) in plasma and cell lysate, after in situ complexation with 1-naphthol. The size-exclusion HPLC column packed with TSK 3000 SW gel , was equilibrated with an eluent mixture composed of methanol and pur ified water (2:98, v/v) containing 10(-4) M 1-naphthol as a fluorophor e. The detection is based on fluorescence enhancement caused by the fo rmation of inclusion complexes and was performed at 290 and 360 nm for excitation and emission, respectively. The method involved a simple t reatment of the samples with chloroform. Daunorubicin was used as inte rnal standard. Limits of quantitation were 0.8 mu M in plasma and 0.5 mu M in cell lysate. Detection limits of 0.5 mu M (50 pmol) and 0.3 mu M (30 pmol) were obtained for MEBCD in the two media, respectively. L inear detection response was obtained for concentrations ranging from 1 to 100 mu M in plasma and cell lysate. Recovery from plasma proved t o be more than 40%. Precision, expressed as C.V. was in the range of 4 to 11%. Accuracy ranged from 89 to 105%.