Measurement of unbound cocaine in blood, brain and bile of anesthetized rats using microdialysis coupled with liquid chromatography and verified by tandem mass spectrometry.
To investigate the disposition of unbound cocaine in the rat blood, brain and bile, we demonstrate an in vivo multiple sampling microdialysis system coupled with liquid chromatography for cocaine assay and verified by tandem mass spectrometry. Three microdialysis probes were concurrently inserted into the jugular vein, bile duct and brain striatum of each anesthetized rat. After a period of 2 h post-surgical stabilization, cocaine (10 mg kg(-1)) was administered through the femoral vein. Separation of unbound cocaine from various biological fluids was applied to a reversed-phase C(18) column (250 x 4.6 mm I.D., 5 microm). The mobile phase consisted of acetonitrile--10 mm potassium dihydrogen phosphate buffer (25:75, v/v, pH 4.0) and 0.8% diethylamine at a flow rate of 1 mL min(-1). The UV detector wavelength was set at 235 nm. The results indicate that cocaine penetrates the blood--brain barrier with a rapid distribution. However, unbound cocaine in the bile dialysate was not detectable in the UV detection. We therefore use LC--tandem mass spectrometry to detect the bile fluid after cocaine administration (3 mg kg(-1), i.v.). The results indicate that cocaine goes through hepatobiliary excretion.
Chen
Yen-Fei
YF
Institute of Traditional Medicine, National Yang-Ming University, Taipei, Taiwan.
Chang
Chun-Hao
CH
Wang
Shau-Chun
SC
Tsai
Tung-Hu
TH
D000768:Anesthesia, General; D000818:Animals; D001646:Bile / Q000737:chemistry*; D001923:Brain Chemistry; D002698:Chloralose; D002853:Chromatography, Liquid / Q000379:methods; D003042:Cocaine / Q000032:analysis* / Q000097:blood / Q000493:pharmacokinetics; D008297:Male; D013058:Mass Spectrometry / Q000379:methods*; D017551:Microdialysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D014520:Urethane
D002698:Chloralose; D014520:Urethane; D003042:Cocaine