Analytical Sciences

Determination of Anti-virus Drug, Ganciclovir, in Human Serum by HPLC with Precolumn Fluorescence Derivatization Using Phenylglyoxal

Masahiko TSUCHIE,* Shuuji HARA,*  Masahiko KIMURA,* Megumi FUJII,* Nobufumi ONO,* and Masaaki KAI**

*Faculty of Pharmaceutical Sciences, Fukuoka University, Nanakuma, Jonan, Fukuoka 814-0180, Japan
**School of Pharmaceutical Sciences, Nagasaki University, Bunkyo-Machi, Nagasaki 852-8521, Japan

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°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.

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