On-line determination of fluconazole in blood and dermal rat microdialysates by microbore high-performance liquid chromatography.
To study the distribution of fluconazole in the dermis of the rat, on-line microdialysis using double-site sampling coupled with a microbore HPLC system was developed. The chromatographic conditions consisted of a mobile phase of 20 mM diammonium phosphate-acetonitrile (75:25, v/v, pH 7.0) pumped through a microbore C(18) column at 40 microl/min. The eluent was monitored with UV detector with UZ flow cell (30 mm path length) at 210 nm. A microbore 10-port pneumatic valve fitted with two loops of 1 microl was used to collect and directly inject microdialysates from jugular and dermal probes. The retention time was 5.8 min for fluconazole and 10.1 min for its fluorinated analog, UK-54373 used as a retrodialysis marker. The assay was precise, with inter- and intra-assay relative standard deviation values of 0.64 and 0.71%, respectively, and with a good linearity (r=0.999) in the range of 0.15-20 microg/ml with only 1 microl injected onto the column. The LOD and LOQ values for fluconazole were 0.100 and 0.150 microg/ml, respectively. The applicability of the method was demonstrated by studying the disposition of fluconazole in blood and dermis following i.v. bolus at a dose of 10 mg/kg.
Mathy
François-Xavier
FX
Unité de Pharmacie Galénique, Université catholique de Louvain, Av. E. Mounier 73, UCL 73.20, 1200 Brussels, Belgium.
Vroman
Benoît
B
Ntivunwa
Denis
D
De Winne
Ann J
AJ
Verbeeck
Roger K
RK
Préat
Véronique
V
D000818:Animals; D000935:Antifungal Agents / Q000032:analysis* / Q000097:blood; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D015725:Fluconazole / Q000032:analysis* / Q000097:blood; D008297:Male; D017551:Microdialysis; D051381:Rats; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012867:Skin / Q000737:chemistry*
D000935:Antifungal Agents; D015725:Fluconazole