Capillary electrochromatography of complex plasma matrix on a C18/SCX column using UV-Vis and mass spectrometric detection

被引:19
作者
Spikmans, V [1 ]
Lane, SJ
Smith, NW
机构
[1] Univ London Imperial Coll Sci Technol & Med, Zeneca SmithKline Beecham Ctr Analyt Sci, London SW7 2AY, England
[2] Glaxo Wellcome Res & Dev Ltd, Stevenage SG1 2NY, Herts, England
关键词
capillary electrochromatography; mass spectrometry; mixed mode columns; plasma analysis;
D O I
10.1007/BF02490690
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Separations of compounds in plasma were performed on a Hypersil Duet C-18/SCX capillary eleetrochromatography (CEC) column, utilizing an automated injection system combined with a short in-house designed and fabricated micro-electrospray CEC mass spectrometer interface. Protein precipitation was used prior to the CEC separations. More than two hundred separations of the corticosteroids Dexamethasone and Betamethasone 17-valerate, and Fluticasone Propionate in complex plasma matrix were pet-formed on a single column under isocratic conditions. The method demonstrated good reproducibility, selectivity, sensitivity and high efficiencies. Linear calibration with good correlation was typical. Estimated detection limits in the low micromolar and nanomolar range for all compounds were obtained using UV-Vis absorbance (UV) and Electrospray Mass Spectrometric (ES/MS) detection respectively. Efficiencies for all compounds were typically 87,500 plates on a 25 cm column (350,000 plates m(-1)) and increased with the number of plasma samples injected, up to 250,000 plates per column (1,000,000 plates m(-1)). These very high observed plate counts may be artificially enhanced by the inadequate scan possibilities of the MS over very narrow chromatographic peaks.
引用
收藏
页码:18 / 24
页数:7
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