The design of an on-line semi-preparative LC-SPE-NMR system for trace analysis.
This paper reports the design of an on-line semi-preparative LC-SPE-NMR system and its use in the structural analysis of mixture components at the 0.02-1% level. The combination provides at least a five fold mass sensitivity increase over that obtained from typical analytical LC-SPE systems and a >30-fold total NMR sensitivity enhancement over analysis by LC-NMR. This is accomplished by using a novel on-line device to store, dilute (1-100-fold) and deliver (at an optimized flow-rate) the isolated component of interest to an SPE trap unit. The SPE unit consists of two cartridges connected in parallel to increase the overall SPE capacity and also to decrease the flow-rate through each trap for enhanced trapping efficiency. As the coupling of semi-preparative LC with NMR (through SPE) is well matched in terms of optimal mass loading for both techniques, only one LC-SPE cycle is required to enrich a 50 microg ml(-1) component (1% in a 5 mg ml(-1) mixture) for the acquisition of heteronuclear (1)H-(13)C NMR data using a conventional NMR flow probe. Furthermore, analytes at the 0.02% level (approximately 1 microg ml(-1)) can be studied using 2D (1)H NMR techniques if peak cuts from replicate sample injections (> or =3) are accumulated into the storage/dilution unit and the resulting solution processed by just one SPE trap and elute cycle.
Xu
Feng
F
Analytical Research and Development, Bristol-Myers Squibb Pharmaceutical Research Institute, Wallingford, Connecticut 06492, USA. feng.xu@bms.com
Alexander
Anthony J
AJ
D002853:Chromatography, Liquid / Q000379:methods*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008832:Microchemistry; D004364:Pharmaceutical Preparations / Q000032:analysis* / Q000737:chemistry; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D004364:Pharmaceutical Preparations