Determination of Isepamicin in the Eluates from the Dried Blood Spots on Filter Paper for Monitoring of Blood Levels in a Guinea-pig

In general, collection of serial blood samples from small experimental animals is difficult in terms of sampling site and operation technique. To overcome these problems, a simple reproducible method has been improved by the use of filter papers. Whole blood obtained by venipuncture from the ear vein of guinea-pig was spotted onto a filter paper. Isepamicin in the dried blood spot was extracted with 0.5 M Na2HPO4 buffer by incubation and determined by fluorescence polarization immunoassay. Linearity was established over the range of 5-150 μg/ml by using only 100 μl of whole blood. Consequently its accuracy and precision were good, with mean coefficient of variation of less than 5%. The method described here correlates well with a conventional sampling method and could be used for the pre-clinical study of isepamicin blood levels of individual guinea-pigs. This method is suitable for the simulataneous measurement of aminoglycoside antibiotics or physiological parameters after the administration of aminoglycoside antibiotics for the pharmacokinetics study.