High-performance liquid chromatography coupled to ion spray mass spectrometry for the determination of colchicine at ppb levels in human biofluids.
An original method based upon high-performance liquid chromatography coupled to ion spray mass spectrometry (HPLC-ISP-MS) has been developed for the identification and quantification of colchicine (COL) in human blood, plasma or urine. After single-step liquid-liquid extraction by dichloromethane at pH 8.0 using tofisopam (TOF) as an internal standard, solutes are separated on a 5-microns C18 Microbore (Alltech) column (250 x 1.0 mm, I.D.), using acetonitrile-2 mM NH4COOH, pH 3 buffer (75: 25, v/v) as the mobile phase (flow-rate 50 microliters/min). Detection is done by a Perkin-Elmer Sciex API-100 mass analyzer equipped with a ISP interface (nebulizing and curtain gas: N2, quality U; main settings: ISP, +4.0 kV; OR, +50 V; Q0, -10 V; Q1, -13 V; electron multiplier, +2.2 kV); MS data are collected as either total ion current (TIC, m/z 100-500 or 380-405), or selected ion monitoring (SIM) at m/z 400 and 383 for COL and TOF, respectively. COL mass spectrum shows a prominent molecular ion [M + H]+ at m/z 400. Increasing OR potential fails to provide a significant fragmentation. Retention times are 2.70 and 4.53 min for COL and TOF, respectively. The quantification method shows a good linearity (r = 0.998) over a concentration range from 5 to 200 ng/ml. The lower limit of detection in SIM mode is 0.6 ng/ml COL, making the method convenient for both clinical and forensic purposes.
Tracqui
A
A
Institut de Médecine Légale, Strasbourg, France.
Kintz
P
P
Ludes
B
B
Rougé
C
C
Douibi
H
H
Mangin
P
P
D014151:Anti-Anxiety Agents; D001569:Benzodiazepines / Q000097:blood / Q000652:urine; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003078:Colchicine / Q000097:blood* / Q000652:urine*; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D012015:Reference Standards; D012680:Sensitivity and Specificity
D014151:Anti-Anxiety Agents; D001569:Benzodiazepines; D003078:Colchicine; C012582:tofisopam