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Exploitation of the modulation freedom by on-column peak refocusing in TRLC×RPLC : speeding up 2D-LC via combined temperature and flow rate gradients

Kristina Wicht, Mathijs Baert, Sonja Schipperges, Norwin von Doehren, André de Villiers and Frederic Lynen (UGent)
(2022) 17th International Symposium on Hyphenated Techniques in Chromatography and Separation Technology (HTC-17), Abstracts.
Author
Organization
Abstract
In recent years, TRLC was introduced as a separation mode that offers interesting features when implemented in the first dimension (1D) of 2D-LC. The separation mode is based on the coupling of stimuli-responsive polymer to the silica supporting material. Commonly, poly(N-isopropyl acrylamide) is used, which depicts increasing retention of hydrophobic compounds for rising temperatures. One of the major benefits of this type of stationary phase is that these effects occur in water, thus allowing for the usage of purely aqueous mobile phases. In combination with RPLC, this establishes a great advantage in terms of robust modulation, as full solute refocusing at the 2D column head is obtained without methodical complexity for solutes which can be retained in RPLC. With more commonly used LC×LC combinations, this transfer of 1D effluent is often hampered due to the peak distorting effects of transferring high organic content. It was shown in TRLC×RPLC, that the volume transferred can be chosen freely, tested for transfer volumes of 10-2000 µL without peak broadening. The combination of the analyte refocusing and the resulting almost unlimited transferable injection volumes in 2D also paves the way for the implementation of flow rate gradients in the 1D. In this way, in this work, the possibilities offered by TRLC in 2D-LC are explored to reduce analysis times while maintaining or improving the overall method performance. This can be done via the usage of single-component type gradients (comprising either a decreasing temperature or increasing flow rate gradient) or combined dual-component gradients (combining temperature and flow rate gradients). It is shown that a change in flow rate (e.g. from 100 - 400 µL/min) and the consequent increase in modulation volume has little to no effect on the 2D separation. The possibilities of the approach are demonstrated through the separation of representative mixtures of small pharmaceuticals and food additives.

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MLA
Wicht, Kristina, et al. “Exploitation of the Modulation Freedom by On-Column Peak Refocusing in TRLC×RPLC :  Speeding up 2D-LC via Combined Temperature and Flow Rate Gradients.” 17th International Symposium on Hyphenated Techniques in Chromatography and Separation Technology (HTC-17), Abstracts, 2022.
APA
Wicht, K., Baert, M., Schipperges, S., von Doehren, N., de Villiers, A., & Lynen, F. (2022). Exploitation of the modulation freedom by on-column peak refocusing in TRLC×RPLC :  speeding up 2D-LC via combined temperature and flow rate gradients. 17th International Symposium on Hyphenated Techniques in Chromatography and Separation Technology (HTC-17), Abstracts. Presented at the 17th International Symposium on Hyphenated Techniques in Chromatography and Separation Technology (HTC-17), Ghent, Belgium.
Chicago author-date
Wicht, Kristina, Mathijs Baert, Sonja Schipperges, Norwin von Doehren, André de Villiers, and Frederic Lynen. 2022. “Exploitation of the Modulation Freedom by On-Column Peak Refocusing in TRLC×RPLC :  Speeding up 2D-LC via Combined Temperature and Flow Rate Gradients.” In 17th International Symposium on Hyphenated Techniques in Chromatography and Separation Technology (HTC-17), Abstracts.
Chicago author-date (all authors)
Wicht, Kristina, Mathijs Baert, Sonja Schipperges, Norwin von Doehren, André de Villiers, and Frederic Lynen. 2022. “Exploitation of the Modulation Freedom by On-Column Peak Refocusing in TRLC×RPLC :  Speeding up 2D-LC via Combined Temperature and Flow Rate Gradients.” In 17th International Symposium on Hyphenated Techniques in Chromatography and Separation Technology (HTC-17), Abstracts.
Vancouver
1.
Wicht K, Baert M, Schipperges S, von Doehren N, de Villiers A, Lynen F. Exploitation of the modulation freedom by on-column peak refocusing in TRLC×RPLC :  speeding up 2D-LC via combined temperature and flow rate gradients. In: 17th International Symposium on Hyphenated Techniques in Chromatography and Separation Technology (HTC-17), Abstracts. 2022.
IEEE
[1]
K. Wicht, M. Baert, S. Schipperges, N. von Doehren, A. de Villiers, and F. Lynen, “Exploitation of the modulation freedom by on-column peak refocusing in TRLC×RPLC :  speeding up 2D-LC via combined temperature and flow rate gradients,” in 17th International Symposium on Hyphenated Techniques in Chromatography and Separation Technology (HTC-17), Abstracts, Ghent, Belgium, 2022.
@inproceedings{8725918,
  abstract     = {{In recent years, TRLC was introduced as a separation mode that offers interesting features when implemented in the first dimension (1D) of 2D-LC. The separation mode is based on the coupling of stimuli-responsive polymer to the silica supporting material. Commonly, poly(N-isopropyl acrylamide) is used, which depicts increasing retention of hydrophobic compounds for rising temperatures. One of the major benefits of this type of stationary phase is that these effects occur in water, thus allowing for the usage of purely aqueous mobile phases. In combination with RPLC, this establishes a great advantage in terms of robust modulation, as full solute refocusing at the 2D column head is obtained without methodical complexity for solutes which can be retained in RPLC. With more commonly used LC×LC combinations, this transfer of 1D effluent is often hampered due to the peak distorting effects of transferring high organic content. It was shown in TRLC×RPLC, that the volume transferred can be chosen freely, tested for transfer volumes of 10-2000 µL without peak broadening. The combination of the analyte refocusing and the resulting almost unlimited transferable injection volumes in 2D also paves the way for the implementation of flow rate gradients in the 1D. In this way, in this work, the possibilities offered by TRLC in 2D-LC are explored to reduce analysis times while maintaining or improving the overall method performance. This can be done via the usage of single-component type gradients (comprising either a decreasing temperature or increasing flow rate gradient) or combined dual-component gradients (combining temperature and flow rate gradients). It is shown that a change in flow rate (e.g. from 100 - 400 µL/min) and the consequent increase in modulation volume has little to no effect on the 2D separation. The possibilities of the approach are demonstrated through the separation of representative mixtures of small pharmaceuticals and food additives.}},
  author       = {{Wicht, Kristina and Baert, Mathijs and Schipperges, Sonja and von Doehren, Norwin and de Villiers, André and Lynen, Frederic}},
  booktitle    = {{17th International Symposium on Hyphenated Techniques in Chromatography and Separation Technology (HTC-17), Abstracts}},
  language     = {{eng}},
  location     = {{Ghent, Belgium}},
  title        = {{Exploitation of the modulation freedom by on-column peak refocusing in TRLC×RPLC :  speeding up 2D-LC via combined temperature and flow rate gradients}},
  year         = {{2022}},
}