Development and validation of an HPLC-DAD method for bis(12)-hupyridone and its application to a pharmacokinetic study.
ABSTRACT A rapid and simple method of high performance liquid chromatography with UV detection for the quantification of bis(12)-hupyridone in rat blood has been developed and validated. Chromatographic separation was carried out in an Agilent Extend C(18) 5microm column (length, 250mm; inner diameter, 4.6mm) using a mixture of water-acetonitrile-trifluoroacetic acid (81:19:0.04, v/v/v) as the mobile phase at a flow rate of 1mL/min, with detection at 229nm. The method used for the bis(12)-hupyridone quantification showed linearity for concentration range of 0.1-7.5microg/mL with r(2)=0.9991. The limit of detection and quantification of this method were 0.05microg/mL and 0.1microg/mL, respectively. The intra- and inter-day variations of the analysis were less than 4.22% with standard errors less than 13.3%. The developed method was successfully applied to the pharmacokinetic study of bis(12)-hupyridone after intravenous administration of 5mg/kg and intraperitoneal administration of 10 and 20mg/kg in rats.