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Instrumentation for advanced microseparations in pharmaceutical analysis and proteomics
Authors:Rozing Gerard  Nägele Edgar  Hörth Patric  Vollmer Martin  Moritz Ralf  Glatz Bernd  Gratzfeld-Hüsgen Angelika
Affiliation:

Agilent Technologies GmbH, Pharmaceutical Solutions, P.O. Box 1280, Waldbronn D 76337, Germany

Abstract:
Small-volume chromatographic columns are only able to generate narrow peaks when flow rates, injection volume and instrument components, such as detector, connecting tubing and fittings, are matched to the peak dispersion from the column. Criteria for the proper design of chromatographic instrumentation are therefore derived from a general model on total dispersion. The performance of such a system is then experimentally evaluated from applications run on narrow-bore, small-volume columns. In order to achieve flow rates that match the dimensions of such columns, a new concept for electronic flow control (EFC) is introduced. A theoretical optimization of column efficiency and throughput is discussed and the results verified with practical examples on short, narrow-bore columns packed with small, porous and superficially porous particles. For complex sample mixtures, the concept of peak capacity is introduced and applied to orthogonal separation principles in multiple chromatographic dimensions through column switching techniques.
Keywords:Capillary HPLC   Nanoflow HPLC   Peak dispersion   Electronic flow control   Superficially porous particles   Column switching   Proteomics   Mass spectrometry
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