Determination of kanamycin by electrophoretically mediated microanalysis with in-capillary derivatization and UV detection.
An electrophoretically mediated microanalysis (EMMA) approach, used to perform on-line chemistry between two small molecules, has been characterized and optimized. The plug-plug type EMMA method involved electrophoretic mixing and subsequent reaction of nanoliter plugs of kanamycin-containing samples and 1,2-phthalic dicarboxaldehyde and mercaptoacetic acid within the confines of the capillary column, which acts as a microreactor. Analyses were performed by pressure-injecting a plug of kanamycin sandwiched in two reagent plugs. A potential of 375 Vcm(-1) was then applied to electrophoretically mix the two reactants, and an incubation time of up to 5 min allowed the reaction to proceed prior to the application of a separation potential of 588 Vcm(-1). UV detection was at 335 nm. The background electrolyte was 30 mM sodium tetraborate at pH 10.0, containing 16% of methanol. The method was validated in terms of linearity, limits of quantitation and detection, and precision. The method allows determination of kanamycin in bulk samples as a fully automated procedure.
Kaale
Eliangiringa
E
Laboratory for Pharmaceutical Chemistry and Drug Analysis, K. U. Leuven, Leuven, Belgium.
Van Schepdael
Ann
A
Roets
Eugène
E
Hoogmartens
Jos
J
D019075:Electrophoresis, Capillary / Q000379:methods*; D007612:Kanamycin / Q000031:analogs & derivatives / Q000032:analysis* / Q000737:chemistry; D008832:Microchemistry / Q000379:methods*; D010795:Phthalic Acids / Q000737:chemistry; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013056:Spectrophotometry, Ultraviolet / Q000379:methods*
C420844:1,2-phthalic dicarboxaldehyde; D010795:Phthalic Acids; D007612:Kanamycin