Performance Assessment of the Waters Breeze QS HPLC System GPC Applications

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