Fluorometric Determination of Sisomicin, an Aminoglycoside Antibiotic, in Dried Blood Spots on Filter Paper by Reversed-Phase High-Performance Liquid Chromatography with Pre-column Derivatization

Abstract

A simple method for the determination of sisomicin (SISO) in dried blood spots (DBS) on filter paper has developed. SISO in DBS was recovered most effectively by ultrasonication in 0.05M KH₂PO₄-borate buffer (pH 9.0). The eluates from the DBS were treated with a carboxymethyl-Sephadex column, followed by determination be reversed-phase high-performance liquid chromatography after pre-column fluorescence derivatization using o-phthalaldehyde and β-mercaptopropionic acid in 0.05M KH₂PO₄-borate buffer (pH 9.0). The detection limit of SISO was below 0.05μg per ml of whole blood (100μl). The method permits a simple blood collection method for monitoring of aminoglycoside antibiotics at therapeutic levels.