Capillary electrophoresis with laser-induced fluorescence: a routine method to determine moxifloxacin in human body fluids in very small sample volumes.
The feasibility of capillary electrophoresis with HeCd laser-induced fluorescence detection as a validated routine method for bioanalytical analysis is reported. Method evaluation, validation and results of the determination of moxifloxacin (BAY 12-8039), a new antimicrobially active 8-methoxy-quinolone, in plasma and microdialysate are described. After a one step sample preparation the samples can be injected directly into the capillary. The volume of microdialysate and plasma, respectively, needed for more than 50 injections is only 10 microl and 20 microl. Total run time is less than 7 min using a 27 cm capillary on commercial instrumentation. An analysis time of less than 1 min was shown to be possible, however it could not be used routinely since appropriate instrumentation was not available. Evaluation is based on the relative corrected peak area (analyte/I.S.). The method's dynamic range comprises three orders of magnitude (plasma: 2.5-5000 microg/l; microdialysate: 5-5000 microg/l). Validation according to international guidelines yielded data on accuracy and precision of the method throughout the entire working range of inter-day precision: plasma <6%, microdialysate <5% and inter-day accuracy: plasma <2%, microdialysate <4%. The crossvalidation with an existing HPLC method utilizing clinical study samples shows linear correlation. In view of its adequate sensitivity and high selectivity capillary electrophoresis with laser-induced fluorescence is a very versatile tool in pharmacokinetic studies of quinolones, especially in situations with limited sample volumes: e. g. pediatrics, patients at risk, animal-, microdialysis- and tissue-kinetic studies. Validation parameters and other features, like high sample throughput and robustness, are comparable to or even better than HPLC. Further necessary improvements of the capillary electrophoresis with laser-induced fluorescence instrumentation (autosampler, vials, parallel capillaries) and its use in bioanalytical routine analysis are discussed.
Möller
J G
JG
Institute of Clinical Pharmacology, Clinical Pharmacokinetics, Bayer AG, Wuppertal, Germany.
Stass
H
H
Heinig
R
R
Blaschke
G
G
D000890:Anti-Infective Agents / Q000032:analysis* / Q000097:blood; D001372:Aza Compounds; D001826:Body Fluids / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid; D019075:Electrophoresis, Capillary / Q000379:methods*; D005453:Fluorescence; D024841:Fluoroquinolones; D006801:Humans; D007834:Lasers; D008297:Male; D008832:Microchemistry; D017551:Microdialysis; D000077266:Moxifloxacin; D011786:Quality Control; D011804:Quinolines; D012680:Sensitivity and Specificity; D013053:Spectrophotometry
D000890:Anti-Infective Agents; D001372:Aza Compounds; D024841:Fluoroquinolones; D011804:Quinolines; D000077266:Moxifloxacin