Improved and simplified liquid chromatographic assay for adefovir, a novel antiviral drug, in human plasma using derivatization with chloroacetaldehyde.
A rapid and simplified chromatographic assay is reported for the quantification of adefovir (PMEA) utilizing derivatization with chloroacetaldehyde. Adefovir is isolated from plasma using protein precipitation with trichloroacetic acid; next, the fluorescent 1,N6-etheno derivative is directly formed at 98 degrees C in the buffered extract with chloroacetaldehyde. This derivative is analyzed using isocratic ion-pair liquid chromatography and fluorescence detection at 254 nm for excitation and 425 nm for emission. In the evaluated concentration range (10-1000 ng/ml) precisions < or = 5% and accuracies between 95 and 117% were found, using a 0.2-ml volume of plasma. The lower limit of quantification is 10 ng/ml with a intra-assay precision of 16%. The currently reported bioanalytical method is 20-25-fold more sensitive than previously published assays.
Sparidans
R W
RW
Utrecht University, Faculty of Pharmacy, Department of Pharmaceutical Analysis, The Netherlands. r.w.sparidans@pharm.uu.nl
Veldkamp
A
A
Hoetelmans
R M
RM
Beijnen
J H
JH
D000079:Acetaldehyde / Q000031:analogs & derivatives*; D000225:Adenine / Q000031:analogs & derivatives* / Q000097:blood; D000998:Antiviral Agents / Q000097:blood*; D011232:Chemical Precipitation; D002853:Chromatography, Liquid / Q000379:methods*; D004355:Drug Stability; D005456:Fluorescent Dyes; D006801:Humans; D007202:Indicators and Reagents; D013058:Mass Spectrometry; D063065:Organophosphonates; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence; D014238:Trichloroacetic Acid
D000998:Antiviral Agents; D005456:Fluorescent Dyes; D007202:Indicators and Reagents; D063065:Organophosphonates; D014238:Trichloroacetic Acid; C053001:adefovir; C004656:chloroacetaldehyde; D000079:Acetaldehyde; D000225:Adenine