fr
Article scientifique
Anglais

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

Publié dansEuropean journal of pharmaceutics and biopharmaceutics, vol. 66, no. 3, p. 475-482
Date de publication2007
Résumé

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.

Mots-clés
  • Chromatography, High Pressure Liquid
  • Chromatography, Liquid/methods
  • Pharmaceutical Preparations/analysis
Citation (format ISO)
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
Fichiers principaux (1)
Article (Published version)
accessLevelRestricted
Identifiants
ISSN du journal0939-6411
865vues
1téléchargements

Informations techniques

Création15/06/2010 12:09:00
Première validation15/06/2010 12:09:00
Heure de mise à jour14/03/2023 15:30:04
Changement de statut14/03/2023 15:30:04
Dernière indexation15/01/2024 20:03:52
All rights reserved by Archive ouverte UNIGE and the University of GenevaunigeBlack