Use of teicoplanin stationary phase for the enantiomeric resolution of atenolol in human urine by nano-liquid chromatography-mass spectrometry.
Nano-liquid chromatography (nano-LC) was used for the enantiomeric resolution of atenolol employing a teicoplanin modified silica stationary phase prepared in our laboratory. Experiments were carried out in a fused silica capillary of 75 microm i.d. packed with chiral modified silica particles of 5 microm diameter. Separated enantiomers were revealed by on-line UV detector at 205 nm or electrospray-ion-trap mass spectrometer (ESI-MS). Atenolol enantiomers were eluted utilizing a mobile phase with the following composition: 500 mM ammonium acetate pH 4.5/methanol/acetonitrile 1:60:39 (v/v/v) allowing to achieve good enantioresolution in a reasonable analysis time (about 8 min) with a flow rate of about 900 nL/min. After comparing the sensitivity of the nano-LC method using a conventional UV detector for capillary electrophoresis, a zeta cell (3 nL volume) employed in nano-LC and the ion-trap MS the method was validated with the MS detector offering the highest sensitivity (limit of detection (LOD)=50 ng/mL; limit of quantification (LOQ)=400 ng/mL for each atenolol enantiomer). (-)-Psi-Nor-ephedrine was used as the internal standard. The method was successfully applied to the analysis of atenolol enantiomers present in human urine samples of a patient under atenolol therapy.
D'Orazio
Giovanni
G
Institute of Chemical Methodologies, Consiglio Nazionale delle Ricerche, Area della Ricerca di Roma I, Via Salaria Km 29,300, 00016 Monterotondo Scalo, Roma, Italy.
Fanali
Salvatore
S
D000319:Adrenergic beta-Antagonists / Q000652:urine*; D000465:Algorithms; D001262:Atenolol / Q000652:urine*; D002138:Calibration; D002853:Chromatography, Liquid; D006801:Humans; D007202:Indicators and Reagents; D013058:Mass Spectrometry; D036103:Nanotechnology; D021241:Spectrometry, Mass, Electrospray Ionization; D013056:Spectrophotometry, Ultraviolet; D013237:Stereoisomerism; D017334:Teicoplanin / Q000737:chemistry*
D000319:Adrenergic beta-Antagonists; D007202:Indicators and Reagents; D001262:Atenolol; D017334:Teicoplanin