Determination of anticancer drug vitamin K3 in plasma by high-performance liquid chromatography

Synthetic vitamin K3 (VK3, 2-methyl-1,4-naphthoquinone, or menadione) has been found to exhibit antitumor activity against various human cancer cells at relative high dose. Parallel to our study on the mechanism of VK3 action and for future clinical trials in Taiwan, we developed a simple, sensitive and accurate high-performance liquid chromatographic method for the determination of VK3 in biological fluids. VK3 was extracted from the plasma samples with n-hexane. The chromatographic separation employed an ODS analytical column (5 microns, 250 x 4.6 mm I.D.) with a mobile phase of methanol-water (70:30, v/v) and UV detection at 265 nm. On completely drying of the extraction solution, n-hexane, by a stream of nitrogen, menadione was lost to a great extent. Methanol (70%, 200 microliters) was added to the extraction solvent after extraction and centrifugation to prevent the loss of menadione. The absolute recovery was 82.4 +/- 7.69% (n = 7). The within-day and between-day calibration curves of VK3 in plasma in the ranges of interest (0.01-10.00 micrograms/ml; 0.01-5.00 micrograms/ml) showed good linearity (r > 0.999) and acceptable precision. The limit of quantitation of VK3 was 10 ng/ml in plasma. This method has been successfully applied to a pilot pharmacokinetic study of VK3 in rabbits receiving an intravenous high-dose bolus injection of 75 mg menadiol sodium diphosphate (Synkayvite). The pharmacokinetic properties of menadione could be described adequately by an open two-compartment model. The mean half-life of menadiol (transformation to menadione) was 2.60 +/- 0.12 min.(ABSTRACT TRUNCATED AT 250 WORDS)