28 August / September 2019
ease of use for medicinal chemists, reduced capital and consumable costs and reduced fraction volumes. The main advantage being that chemists now have the ability to purify extremely polar samples by use of HILIC flash which is readily available at their own fume hood, instead of having to submit samples for HILIC semi-prep or normal phase, SFC separations. HILIC chromatography on flash has been demonstrated to be reproducible. The ability to purify nucleosides and nucleotides is impressive and demonstrates how easy this methodology is to adapt to medicinal chemistry programs where polar compound separations is a bottleneck. Additionally, the use of alternative stationary phases other than bare silica has been discussed for analytical HILIC and these phases are also available in flash chromatography cartridge form.
References
1. A.J. Alpert, Hydrophilic-interaction chromatography for the separation of peptides, nucleic acids and other polar compounds. J. Chromatography, 499, 177 (1990)
2. N.P. Dinh, T. Jonsson and K. Irgum, Probing the interaction mode in hydrophilic interaction chromatography. J. Chromatography A, 1218, 5880-5991 (2011)
3. S. Cubbon, T. Bradbury, J. Wilson and J. Thomas-Oates, Hydrophilic interaction chromatography for mass spectrometric metabonomic studies of urine. Anal. Chem, 79, 8911-8918 (2007)
4. M. Isokawa, T. Kanamori, T. Funatsu and M. Tsunoda, Recent advances in hydrophilic interaction chromatography for quantitative analysis of endogenous and pharmaceutical compounds in plasma samples. Bioanalysis 6(18), 2421- 2439 (2014)
5. J.P. Danaceau, E.E. Chambers and K.J. Fountain, Hydrophilic Interaction Chromatography (HILIC) for LCMS/ MS of monoamine neurotransmitters Bioanalysis 4(7), 783-794 (2012)
6. K. Spagou, H. Tsoukali, N. Raikos, H. Gika, I. Wilson and G. Theodoridis, Hydrophilic Interaction Chromatography coupled to MS for metabonomic/ metabolomic studies, J. Sep. Sci. 33, 716-727 (2010)
7. J. Heaton, N. Gray, D.A. Cowan, R. Plumb, C. Legido-Quigley and N.W. Smith, Comparison of reversed-phase and hydrophilic interaction liquid chromatography for the separation of ephedrines, J. Chromatography A, 1228, 329-337 (2012)
8. J.C. Heaton and D.V. McCalley, Some factors that can lead to poor peak shape in hydrophilic interaction chromatography, and possibilities for
their remediation, J. Chromatography A, 1427, 37-44 (2016)
9. W. Yu, H. Jin, A. Shen, L. Deng, J. Shi, X. Xue, Y. Guo, Y. Liu and X. Liang, Purification of high-purity glycyrrhizin from licorice using hydrophilic interaction solid phase extraction coupled with preparative reversed- phase liquid chromatography, J. Chromatography B, 1040, 47-52 (2017)
10. G. Liu, B. Luan, G. Liang, L. Xing, L. Huang, C. Wang and Y. Xu, Isolation and identification of four major impurities in capreomycin, J. Chromatography A, 1571, 155-164 (2018)
11. G. Jaffuel, L. Chappuis, D. Guillarme, T.C.J Turlings and G. Glauser, Improved separation by at-column in preparative hydrophilic interaction chromatography, J. Chromatography A, 1532, 136-143 (2018)
12. J. Reilly, Q. Liu, C. Howson, K. Boss, B. Shin, M. Palmermo, J. Larrow, M. Qing and T. Mann, Method Development for Reproducible Flash Purification of Pharmaceuticals by UHPLC and HPLC, Chromatography Today, Nov/Dec issue, 2-5, 2016
13. C. Zhang, Z. Liu, X. Liu, L. Wei, Y. Liu, J. Yu and L. Sun, Targeted metabolic analysis of nucleotides and identification of biomarkers associated with cancer in cultured cell models, Acta Pharmaceutica Sinica B, 3(4), 254-262 (2013)
The Advantages of Coupling GCxGC with QTOF Detection in MS/MS Mode
An application note from JSB studies the feasibility of coupling GCxGC with QTOF detection in MS/MS mode.
GC-QTOF is a valuable technique for challenging applications that demand confident identification in highly complex matrices, such as, metabolomics and pesticides analysis. Comprehensive two-dimensional gas chromatography (GC×GC) is the chromatographic method of choice to unravel high sample complexity. Compared to classic GC, GC×GC grants superior separation power and unmatched peak capacity by coupling two different separation mechanism in a single analysis.
The results of the study show the practicality of operating the QTOF in MS/ MS mode also coupled to GCxGC. This feature can be a powerful profiling tool to complement two-dimensional chromatograms and HRMS and their
excellent selectivity in order to achieve even more confident identification and determine more structures of unknowns.
The study concluded that by using the Agilent 7250 QTOF detector, high-resolution, accurate mass, soft ionisation and MS/MS capability was ensured. These features are generally used in GC-QTOF for advanced identity confirmation and structure elucidation. It was possible to use GC×GC with the QTOF in MS/MS mode, and by careful optimisation, good MS/MS data, also for the sharp modulated peaks, was obtained. This technique can be a very powerful tool for the identification of targets and unknowns with enhanced confidence and selectivity.
More information online:
ilmt.co/PL/v0Jm and
ilmt.co/PL/AREZ
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