Author/Authors :
Szemلn، نويسنده , , Julianna and Gerlَczy، نويسنده , , Andrea and Csabai، نويسنده , , Katalin and Szejtli، نويسنده , , Jَzsef and Kis، نويسنده , , Georg L and Su، نويسنده , , Ping and Chau، نويسنده , , Raymond Y and Jacober، نويسنده , , Albert، نويسنده ,
Abstract :
A high-performance size exclusion chromatographic method with analyte enhanced fluorescence detection is described for the analysis of 2-hydroxypropyl-γ-cyclodextrin (HPGCD) in different biological fluids. The principle of detection was the in situ complexation of 8-anilinonaphthalene-1-sulfonic acid (ANS) by HPGCD. When HPGCD eluted from the column the increased fluorescence was measured at excitation and emission wavelengths of 270 and 512 nm, respectively. Solid-phase extraction cleanup and concentration of samples resulted in higher than 78% recovery of HPGCD for each of the studied biological fluids. Some important details of the method development as well as the validation of the method for rabbit plasma, rabbit aqueous humour, monkey plasma and monkey urine are given. The limits of quantification varied between 1 and 10 nmol/ml (correspond to 1.5–15 μg/ml) depending on the biological matrix used. The method was successfully adapted in another laboratory proving that HPGCD had not absorbed into aqueous humour and plasma after topical application of HPGCD containing eye drop in rabbits.
