Operating conditions in preparative medium pressure liquid chromatography (MPLC).II. Influence of solvent strength and flow rate of the mobile phase, capacity and dimensions of the column

G. C. Zogg, S. Nyiredy, O. Sticher

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Abstract

This study describes the influence of some of the most important operating parameters in preparative medium pressure liquid chromatography (MPLC). Apolar test compounds with closely related structures were used for the experiments. For all separations the columns were dry-packed with the aid of vacuum and nitrogen overpressure and then equilibrated with n-hexane. The variation of the solvent strengths and sample loads gave a linear correlation. In view of the high resolutions also obtained in preparative MPLC separations, sample loads of 3-10 mg extract/g support can be used for the isolation of closely related compounds, with high purity (≥95%). A wide range of dimensions of MPLC glass column was tested to study the influence of column diameter and length. The diameter was varied from 26 to 49 cm, the length between 173 and 920 cm and the volume between 102 and 1740 ml. The resolution between all compounds to be separated indicated a linear correlation for the applied column dimensions. A pronounced decrease in the net retention times could be observed on increasing the flow rate but the flow rate does not exert any pronounced influence on the resolution, when working in a range where the backpressure does not exceed 20 bar and keeping all other parameters constant.

Original languageEnglish
Pages (from-to)2049-2065
Number of pages17
JournalJournal of Liquid Chromatography
Volume12
Issue number11
Publication statusPublished - Sep 1989

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Liquid chromatography
Liquid Chromatography
Flow rate
Pressure
Vacuum
Glass
Nitrogen
Experiments

ASJC Scopus subject areas

  • Molecular Medicine

Cite this

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AU - Sticher, O.

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N2 - This study describes the influence of some of the most important operating parameters in preparative medium pressure liquid chromatography (MPLC). Apolar test compounds with closely related structures were used for the experiments. For all separations the columns were dry-packed with the aid of vacuum and nitrogen overpressure and then equilibrated with n-hexane. The variation of the solvent strengths and sample loads gave a linear correlation. In view of the high resolutions also obtained in preparative MPLC separations, sample loads of 3-10 mg extract/g support can be used for the isolation of closely related compounds, with high purity (≥95%). A wide range of dimensions of MPLC glass column was tested to study the influence of column diameter and length. The diameter was varied from 26 to 49 cm, the length between 173 and 920 cm and the volume between 102 and 1740 ml. The resolution between all compounds to be separated indicated a linear correlation for the applied column dimensions. A pronounced decrease in the net retention times could be observed on increasing the flow rate but the flow rate does not exert any pronounced influence on the resolution, when working in a range where the backpressure does not exceed 20 bar and keeping all other parameters constant.

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