Advanced search
2 files | 7.00 MB Add to list

Enhanced sensitivity in comprehensive liquid chromatography : overcoming the dilution problem in LC × LC via temperature-responsive liquid chromatography

(2022) ANALYTICAL CHEMISTRY. 94(48). p.16728-16737
Author
Organization
Project
Abstract
In comprehensive two-dimensional liquid chromatography (LC x LC), solvents of high eluotropic strength are frequently used in the first dimension (1D), which lead to peak broadening in the second dimension (2D). In the majority of the current LC x LC column combinations, analytes are less than optimally refocused upon transfer to the second column, which negatively affects sensitivity. Furthermore, the typical combination of 1 or 2.1 mm columns in the 1D paired with a 3 mm (or broader) column in the 2D leads to at least a 9-or 4-fold dilution and a corresponding loss of sensitivity when using concentration-sensitive detectors. This occurs due to the enhanced radial dilution of the analytes in a broader column, while the sensitivity problem is further exacerbated in LC x LC due to the high flow operated 2D. In this paper, we introduce a solution to neutralize and inverse this dilution problem through a reconcentrating solution using temperature-responsive liquid chromatography (TRLC) in the 1D, which is a purely aqueous separation mode. Full solute refocusing at the 2D column head is thereby obtained when TRLC is combined with reversed-phase liquid chromatography (RPLC). This is shown for the combination of a 2.1 mm I.D. TRLC column with decreasing RPLC column diameters (3-2.1-1 mm) operated at the same linear velocities, hence a resulting decrease in dilution, respectively. Ultraviolet (UV) and electrospray ionization time-of-flight mass spectrometry (ESI-TOF-MS) detection were used to determine the experimental detection limits. Sensitivity improvements with UV detection were somewhat lower than expected, but represent similar to 1.5-and 3-fold sensitivity enhancement when using a 1 mm I.D. column compared to 2.1 or 3 mm I.D. columns in the 2D, respectively. This is attributed to extra-column dispersion and the poorer performance of 1 mm I.D. columns. A major benefit of the use of 1 mm I.D. columns in the 2D is that it allows split-free coupling of 2D effluent with ESI-MS (at 450 mu L/min), making the coupling robust and simple. When using ESI-MS even better, albeit more variable, sensitivity enhancements were obtained on the narrower columns. The benefits of the methodology are demonstrated for paraben test solutes and for phenolic compounds in a blueberry extract by TRLC x RPLC-UV-ESI-TOF-MS.
Keywords
Analytical Chemistry, MASS-SPECTROMETRY, SEPARATION, OPTIMIZATION, TIME, MODULATION

Downloads

  • (...).pdf
    • full text (Published version)
    • |
    • UGent only
    • |
    • PDF
    • |
    • 3.54 MB
  • AAM.pdf
    • full text (Accepted manuscript)
    • |
    • open access
    • |
    • PDF
    • |
    • 3.46 MB

Citation

Please use this url to cite or link to this publication:

MLA
Wicht, Kristina, et al. “Enhanced Sensitivity in Comprehensive Liquid Chromatography : Overcoming the Dilution Problem in LC × LC via Temperature-Responsive Liquid Chromatography.” ANALYTICAL CHEMISTRY, vol. 94, no. 48, 2022, pp. 16728–37, doi:10.1021/acs.analchem.2c03300.
APA
Wicht, K., Baert, M., Schipperges, S., von Doehren, N., Desmet, G., Van Geem, K., … Lynen, F. (2022). Enhanced sensitivity in comprehensive liquid chromatography : overcoming the dilution problem in LC × LC via temperature-responsive liquid chromatography. ANALYTICAL CHEMISTRY, 94(48), 16728–16737. https://doi.org/10.1021/acs.analchem.2c03300
Chicago author-date
Wicht, Kristina, Mathijs Baert, Sonja Schipperges, Norwin von Doehren, Gert Desmet, Kevin Van Geem, André de Villiers, and Frederic Lynen. 2022. “Enhanced Sensitivity in Comprehensive Liquid Chromatography : Overcoming the Dilution Problem in LC × LC via Temperature-Responsive Liquid Chromatography.” ANALYTICAL CHEMISTRY 94 (48): 16728–37. https://doi.org/10.1021/acs.analchem.2c03300.
Chicago author-date (all authors)
Wicht, Kristina, Mathijs Baert, Sonja Schipperges, Norwin von Doehren, Gert Desmet, Kevin Van Geem, André de Villiers, and Frederic Lynen. 2022. “Enhanced Sensitivity in Comprehensive Liquid Chromatography : Overcoming the Dilution Problem in LC × LC via Temperature-Responsive Liquid Chromatography.” ANALYTICAL CHEMISTRY 94 (48): 16728–16737. doi:10.1021/acs.analchem.2c03300.
Vancouver
1.
Wicht K, Baert M, Schipperges S, von Doehren N, Desmet G, Van Geem K, et al. Enhanced sensitivity in comprehensive liquid chromatography : overcoming the dilution problem in LC × LC via temperature-responsive liquid chromatography. ANALYTICAL CHEMISTRY. 2022;94(48):16728–37.
IEEE
[1]
K. Wicht et al., “Enhanced sensitivity in comprehensive liquid chromatography : overcoming the dilution problem in LC × LC via temperature-responsive liquid chromatography,” ANALYTICAL CHEMISTRY, vol. 94, no. 48, pp. 16728–16737, 2022.
@article{01GQFB2NAF33FHMABWZP9KD3WV,
  abstract     = {{In comprehensive two-dimensional liquid chromatography (LC x LC), solvents of high eluotropic strength are frequently used in the first dimension (1D), which lead to peak broadening in the second dimension (2D). In the majority of the current LC x LC column combinations, analytes are less than optimally refocused upon transfer to the second column, which negatively affects sensitivity. Furthermore, the typical combination of 1 or 2.1 mm columns in the 1D paired with a 3 mm (or broader) column in the 2D leads to at least a 9-or 4-fold dilution and a corresponding loss of sensitivity when using concentration-sensitive detectors. This occurs due to the enhanced radial dilution of the analytes in a broader column, while the sensitivity problem is further exacerbated in LC x LC due to the high flow operated 2D. In this paper, we introduce a solution to neutralize and inverse this dilution problem through a reconcentrating solution using temperature-responsive liquid chromatography (TRLC) in the 1D, which is a purely aqueous separation mode. Full solute refocusing at the 2D column head is thereby obtained when TRLC is combined with reversed-phase liquid chromatography (RPLC). This is shown for the combination of a 2.1 mm I.D. TRLC column with decreasing RPLC column diameters (3-2.1-1 mm) operated at the same linear velocities, hence a resulting decrease in dilution, respectively. Ultraviolet (UV) and electrospray ionization time-of-flight mass spectrometry (ESI-TOF-MS) detection were used to determine the experimental detection limits. Sensitivity improvements with UV detection were somewhat lower than expected, but represent similar to 1.5-and 3-fold sensitivity enhancement when using a 1 mm I.D. column compared to 2.1 or 3 mm I.D. columns in the 2D, respectively. This is attributed to extra-column dispersion and the poorer performance of 1 mm I.D. columns. A major benefit of the use of 1 mm I.D. columns in the 2D is that it allows split-free coupling of 2D effluent with ESI-MS (at 450 mu L/min), making the coupling robust and simple. When using ESI-MS even better, albeit more variable, sensitivity enhancements were obtained on the narrower columns. The benefits of the methodology are demonstrated for paraben test solutes and for phenolic compounds in a blueberry extract by TRLC x RPLC-UV-ESI-TOF-MS.}},
  author       = {{Wicht, Kristina and Baert, Mathijs and Schipperges, Sonja and von Doehren, Norwin and Desmet, Gert and Van Geem, Kevin and de Villiers, André and Lynen, Frederic}},
  issn         = {{0003-2700}},
  journal      = {{ANALYTICAL CHEMISTRY}},
  keywords     = {{Analytical Chemistry,MASS-SPECTROMETRY,SEPARATION,OPTIMIZATION,TIME,MODULATION}},
  language     = {{eng}},
  number       = {{48}},
  pages        = {{16728--16737}},
  title        = {{Enhanced sensitivity in comprehensive liquid chromatography : overcoming the dilution problem in LC × LC via temperature-responsive liquid chromatography}},
  url          = {{http://doi.org/10.1021/acs.analchem.2c03300}},
  volume       = {{94}},
  year         = {{2022}},
}

Altmetric
View in Altmetric
Web of Science
Times cited: