Determination of anti-virus drug, ganciclovir, in human serum by HPLC with precolumn fluorescence derivatization using phenylglyoxal.
A selective and highly sensitive liquid chromatographic method for the determination of ganciclovir (anti-virus drug) in human serum was described. After ganciclovir and acyclovir (internal standard; IS) were extracted with solid-phase extraction cartridge from serum, they were converted into fluorescent derivatives by reaction with phenylglyoxal in a phosphate buffer (pH 5.8) at 20 degrees C for 30 min. The derivatives were separated by reversed-phase column with a mixture of acetonitrile-1 mM phosphate buffer (pH 6.2) (18:82, v/v), and were then detected spectrofluorometrically at 512 nm with excitation at 365 nm. Extraction recoveries were 87.0-91.6% for ganciclovir and 86.8-92.3% for IS. The detection limit for ganciclovir spiked to serum was 5 ng ml-1 serum (306 fmol on column) at a signal-to-noise ratio of three. The accuracy and precision of this method were 7.6% and 5.0% even at low concentration (20 ng ml-1). The within- and between-day variations are lower than 7.6% and 8.1%, respectively.
Tsuchie
M
M
Faculty of Pharmaceutical Sciences, Fukuoka University, Nanakuma, Jonan, Fukuoka 814-0180, Japan.
Hara
S
S
Kimura
M
M
Fujii
M
M
Ono
N
N
Kai
M
M
D000212:Acyclovir / Q000097:blood*; D000998:Antiviral Agents / Q000097:blood* / Q000494:pharmacology; D002624:Chemistry, Clinical / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D015774:Ganciclovir / Q000097:blood*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents / Q000494:pharmacology; D008956:Models, Chemical; D010658:Phenylglyoxal / Q000494:pharmacology*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013997:Time Factors
D000998:Antiviral Agents; D004791:Enzyme Inhibitors; D007202:Indicators and Reagents; D010658:Phenylglyoxal; D015774:Ganciclovir; D000212:Acyclovir