Application Note
Performance Assessment of the Waters
Breeze QS HPLC System GPC Applications
Jennifer Gough,
Neil J. Lander
Waters Corporation
This is an Application Brief and does not contain a detailed Experimental section.
Abstract
This application brief assess the performance of Waters Breeze QS HPLC System for Gel Permeation
Chromatography (GPC).
Benefits
Waters Breeze QS HPLC System, running Empower QuickStart Software, delivers precise flow rate to obtain
accurate, reproducible peaks in GPC tests.
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Waters most affordable GPC analysis using a Breeze QS HPLC System.
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Empower QuickStart (QS) Software: a simplified and intuitive workflow.
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Linear GPC separation to 7 million Daltons with a Waters Ultrahydrogel Linear Column.
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Performance Assessment of the Waters Breeze QS HPLC System GPC Applications
Introduction
Gel permeation chromatography (GPC) requires a high performance liquid chromatography (HPLC) system that
delivers precise flow and reproducible results. In GPC, the retention time is plotted against the log of the
molecular weight. Thus, flow rate variations can affect peak reproducibility. The flow rate for this experiment is
listed in the method parameters (Table 1). The molecular weight of experimental samples is calculated by
comparing of unknown sample data to a calibration curve using narrow molecular weight polyethylene oxide
(PEO) and polyethylene glycol (PEG) calibration standards (Figure 1).
The excellent flow rate precision of Breeze isocratic pumps is demonstrated in an overlay of six injections of a
PEO/PEG mixed molecular weight standard, as shown in Figure 2. Relative standard deviations of the peak
molecular weight (Mp) at <0.5% are obtained on a Waters Breeze QS HPLC System using two linear GPC
columns and a 30-minute run time (Table 2).
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Performance Assessment of the Waters Breeze QS HPLC System GPC Applications
Experimental
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Performance Assessment of the Waters Breeze QS HPLC System GPC Applications
Table 1. Method parameters.
Results and Discussion
Empower QS Software provides easy-to-use fundamental GPC calculations. In this experiment, the narrow
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Performance Assessment of the Waters Breeze QS HPLC System GPC Applications
PEO/PEG standards are used as calibration standards, as well as broad unknowns. The Empower QS Software’s
partial list of reportable peak result fields includes Mn, Mw, Mp, Mv, Mz, Mz+1, and polydispersity (Figure 3).
Relative and modified universal calibration types employing point-to-point and first- through fifth-order fits are
available, and an example of relative calibration is shown in Figure 1.
Figure 1. PEO/PEG calibration.
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Performance Assessment of the Waters Breeze QS HPLC System GPC Applications
Figure 2. Six injections of the PEO/PEG calibration standard.
Table 2. Empower QuickStart data report from six injections of the yellow cap PEO/PEG calibration standard (Mp
= peak molecular weight).
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Performance Assessment of the Waters Breeze QS HPLC System GPC Applications
Figure 3. Calculated peak fields and distribution curve of PEO/PEG calibration standard.
Empower QS has many built-in features that enable sophisticated reporting of data usually done in
spreadsheets. Two popular reporting areas are column plate counts and control charting. By reporting the
column plate counts, the column efficiency can be charted and observed for any changes in performance (Table
3). Control charts can capture upper and lower limits of molecular weight ranges used in a polymer pre-
manufacture notice (PMN) or for charting impurity levels in raw materials (Figure 4).
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Performance Assessment of the Waters Breeze QS HPLC System GPC Applications
Table 3. System suitability features in Empower QS include plate count calculations.
Figure 4. Example of an Empower QS control chart of calibration standard yellow cap peak 4.
Conclusion
Breeze isocratic pumps provide the excellent flow precision necessary for high-quality gel permeation
chromatography.
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Performance Assessment of the Waters Breeze QS HPLC System GPC Applications
All basic GPC calculations can be calculated using the easy-to-use interface included with Empower QS
Software.
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Short analysis time of 30 minutes was demonstrated using two linear columns.
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References
Waters Corporation. Gel Permeation Chromatography (GPC) Capabilities. https://www.waters.com/waters/
library.htm?cid=511436&lid=134697666, August 2014.
1.
Waters Corporation. Empower GPC Software Getting Started Guide. https://www.waters.com/waters/
support.htm?lid=1852970, May 2004.
2.
Waters Corporation. Empower Tips. https://blog.waters.com/category/empower-tips
3.
Featured Products
Breeze QS HPLC <https://www.waters.com/135032418>
Empower 3 Chromatography Data Software <https://www.waters.com/10190669>
720006668, October 2019
©2019 Waters Corporation. All Rights Reserved.
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Performance Assessment of the Waters Breeze QS HPLC System GPC Applications
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Performance Assessment of the Waters Breeze QS HPLC System GPC Applications
