Scientific article

Method transfer for fast liquid chromatography in pharmaceutical analysis: application to short columns packed with small particle. Part I: isocratic separation

Published inEuropean journal of pharmaceutics and biopharmaceutics, vol. 66, no. 3, p. 475-482
Publication date2007

Liquid chromatography (LC) is considered to be the gold standard in pharmaceutical analysis. Today, there is a need for fast and ultra-fast methods with good efficiency and resolution for achieving separations in few minutes or even seconds. The present work describes a simple methodology for performing a successful method transfer from conventional LC to fast and ultra-fast LC. In order to carry out fast separations, short columns (20-50mm) packed with small particles (3.5 and 1.7 microm) were used and their chromatographic performance was compared to that of a conventional column (150 mm, 5 microm). For that purpose, an optimized LC system was employed to limit extra-column volumes which can have a dramatic impact on efficiency and resolution. This paper reports the fundamental equations used for transferring an isocratic chromatographic separation performed with a given column geometry and chemistry to a smaller column packed with similar or identical stationary phase, without influence on chromatographic performance. For this purpose, the flow rate and the injected volume need to be adapted. The effect of column length and particle size reduction on chromatographic resolution and analysis time was described for an isocratic separation. Using the method transfer equations, it is possible to predict the new conditions to be used, for fast and ultra-fast separations. In this work, ultra-fast separations were achieved thanks to a new generation of instrumentation (ultra performance liquid chromatography, UPLC) which uses simultaneously short column packed with sub-2 microm particles and ultra-high pressure (up to 1000 bar). This work demonstrates an analysis time reduction up to a factor 12, compared to a conventional LC separation, without affecting the quality of separation. Therefore, the complete resolution of a pharmaceutical formulation was achieved in only a few seconds.

  • Chromatography, High Pressure Liquid
  • Chromatography, Liquid/methods
  • Pharmaceutical Preparations/analysis
Citation (ISO format)
GUILLARME, Davy et al. Method transfer for fast liquid chromatography in pharmaceutical analysis: application to short columns packed with small particle. Part I: isocratic separation. In: European journal of pharmaceutics and biopharmaceutics, 2007, vol. 66, n° 3, p. 475–482. doi: 10.1016/j.ejpb.2006.11.027
Main files (1)
Article (Published version)
ISSN of the journal0939-6411

Technical informations

Creation06/15/2010 12:09:00 PM
First validation06/15/2010 12:09:00 PM
Update time03/14/2023 3:30:04 PM
Status update03/14/2023 3:30:04 PM
Last indexation10/18/2023 9:15:10 AM
All rights reserved by Archive ouverte UNIGE and the University of GenevaunigeBlack