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Retention characteristics of some antibiotic and anti-retroviral compounds in hydrophilic interaction chromatography using isocratic elution, and gradient elution with repeatable partial equilibration

Heaton, James C.; Smith, Norman W.; McCalley, David V.

Retention characteristics of some antibiotic and anti-retroviral compounds in hydrophilic interaction chromatography using isocratic elution, and gradient elution with repeatable partial equilibration Thumbnail


Authors

James C. Heaton

Norman W. Smith



Abstract

© 2018 Elsevier B.V. The separation of some zwitterionic, basic and neutral antibiotic and antiretroviral compounds was studied using hydrophilic interaction chromatography (HILIC) on bare silica, bonded amide and urea superficially porous phases. The differences in the selectivity and retentivity of these stationary phases were evaluated for compounds with widely different physicochemical properties (logD −3.43 to 2.41 at ww pH 3.0). The mobile phase was acetonitrile-ammonium formate buffered at low ww pH. Compounds containing quinolone and serine groups showed poor peak shapes on all columns, attributed to metal-oxide interactions with system metals. Peak shapes were improved by addition of citrate buffers. Gradient elution, particularly with regard to column equilibration, was also studied due to the large differences in retention factors observed under isocratic conditions. Full equilibration in HILIC was slow for both ionogenic and neutral solutes, requiring as much as ∼40 column volumes. However, highly repeatable partial equilibration, suitable for gradient elution, was achieved in only a few minutes. Pronounced selectivity differences in the separations were shown dependent on the partial equilibration time.

Journal Article Type Article
Publication Date Jan 3, 2019
Journal Analytica Chimica Acta
Print ISSN 0003-2670
Electronic ISSN 1873-4324
Publisher Elsevier
Peer Reviewed Peer Reviewed
Volume 1045
Pages 141-151
APA6 Citation Heaton, J. C., Smith, N. W., & McCalley, D. V. (2019). Retention characteristics of some antibiotic and anti-retroviral compounds in hydrophilic interaction chromatography using isocratic elution, and gradient elution with repeatable partial equilibration. Analytica Chimica Acta, 1045, 141-151. https://doi.org/10.1016/j.aca.2018.08.051
DOI https://doi.org/10.1016/j.aca.2018.08.051
Keywords HILIC, antibiotics, antiretrovirals, gradient elution
Publisher URL http://dx.doi.org/10.1016/j.aca.2018.08.051
Additional Information Additional Information : This is the author's accepted manuscript. The final published version is available here: http://dx.doi.org/10.1016/j.aca.2018.08.051

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