| Abstract: | Procedures for the micro-determination of beta-cyclodextrin (beta-CyD) in plasma were investigated by four methods using high-performance liquid chromatography (HPLC). In methods A and B, underivatized beta-CyD was detected with a refractive index detector and determined by the absolute calibration graph method. An NH2-bonded silica/acetonitrile-water system was used in A and a C18-bonded silica/methanol-water system in B. In method C, the percarbanilate of beta-CyD was separated on a C8-bonded silica column with acetonitrile-water and determined using gamma-CyD as the internal standard with a UV detector at 231 nm. In method D, the per[1-14C]acetate of beta-CyD was fractionated on a silica column with n-hexane-ethanol containing 1% of water and the radioactivity of each fraction was measured with a liquid scintillation counter. gamma-CyD was used as the internal standard. Interfering plasma proteins were removed by centrifugal ultrafiltration with an MPS-1 micro-partition system. Method B was superior to the other methods with respect to ease of sample preparation, sensitivity and time required for analysis. The cumulative amount of beta-CyD in the mesenteric vein absorbed from the rat intestinal lumen after administration of phenobarbital-beta-CyD complex in a closed loop method was determined by the use of method B. |
