Determination of unbound theophylline in rat blood and brain by microdialysis and liquid chromatography.
To investigate the mechanism by which theophylline crosses the blood-brain barrier (BBB) and its disposition, we determined unbound theophylline in rat blood and brain using microbore liquid chromatography coupled with microdialysis. Microdialysis probes were inserted into the jugular vein and the brain striatum of male Sprague-Dawley rats. Then theophylline at dosage of 10 or 30 mg/kg was administered through the femoral vein. Theophylline and dialysates were separated using a microbore phenyl-hexyl column (150 mm x 1 mm, 5 microm). The mobile phase comprised of acetonitrile-methanol-10 mM monosodium phosphate (pH 3.0) (10:20:70, v/v/v). The UV wavelength was set at 270 nm. The concentration-response relationship was linear over a concentration range of 0.05-50 microg/ml; intra-assay and inter-assay precision and accuracy of theophylline fell within 10%. Average in vivo recoveries were 0.74 +/- 0.06 in blood and 0.27 +/- 0.07 in brain with theophylline at concentrations 1, 2 and 5 microg/ml. This biological sampling method thereby allowed the determination of theophylline levels in blood and brain tissues. The disposition of theophylline in the blood and brain tissue suggests that there was rapid exchange and equilibration between the blood and brain system. The drug-drug interaction results indicate that theophylline was able to cross BBB, but that it might not be regulated by p-glycoprotein to the pharmacokinetics of theophylline.
Tsai
Tung-Hu
TH
National Research Institute of Chinese Medicine, 155-1, Li-Nong Street Section 2, Shih-Pai, Taipei 112, Taiwan. thtsai@nricm.edu.tw
Liu
Mei-Chun
MC
D000818:Animals; D001812:Blood-Brain Barrier; D001921:Brain / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D017551:Microdialysis; D051381:Rats; D013806:Theophylline / Q000008:administration & dosage / Q000097:blood / Q000493:pharmacokinetics*
D013806:Theophylline