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Chromatography Software Articles How to Integrate Merged Peaks in OpenLab CDS (ver. 2.x)
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      • How to Create a Multipoint ESTD Calibration Method with Agilent OpenLab CDS (ver 2.x)
      • How to Check the signal-to-noise ratio with Agilent OpenLab CDS (version 2.x)
      • How to Integrate Merged Peaks in OpenLab CDS (ver. 2.x)
      • How to Review Completed Injections in a Running Sequence in Agilent OpenLab CDS (ver 2.x)
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      • How to Hide the Top Triangular Mark of the Peaks in the Intelligent Report with OpenLab CDS (ver. 2.x)
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How to Integrate Merged Peaks in OpenLab CDS (ver. 2.x)

Created by Carlos Vargas Carlos Vargas over 1 year ago | Last modified by Carlos Vargas Carlos Vargas 7 months ago

This Information Applies To: Agilent OpenLab CDS (ver 2.x)


Issue:

How to integrate non-ideally separated peaks starting above the zero baseline in OpenLab CDS


Resolution

Integrating merged peaks is a classical challenge in chromatography in cases where two compounds cannot be fully separated. This post outlines some key steps and references to helpful resources on how to address this problem.

What are merged peaks?

Merged peaks occur when a new peak begins before the end of peak is found. Figure 1 illustrates how the integrator deals with merged peaks.


Figure 1. Merged Peaks Example

How to integrate merged peaks in OpenLab CDS?

  • General recommendation is to optimize the chromatography to fully separate peaks. 

  • In case you cannot avoid coelution, optimize the automated integration to avoid unnecessary use of manual integration, which is prone to human error and variation in interpretation

  • Begin by defining starting integration events, then fine-tune the integration results. (See Figure 2)


Figure 2. Baseline construction

Peak Valley Ratio

The Peak to valley ratio is a measure of quality, indicating how well the peak is separated from other substance peaks. This user-specified parameter is a constituent of advanced baseline tracking mode. 

  • By adjusting the peak to valley ratio, you can decide if two peaks that do not show baseline separation are separated using a drop line or a valley baseline. (See video in Figure 4)

  • The peak-to-valley ratio is calculated using the following equations:

    H1 ≥ H2, Peak valley ratio = H2/Hv
    and
    H1 < H2, Peak valley ratio = H1/Hv;

    where H1=peak height of the first peak, H2=peak height of the second peak, Hv= height of the valley between the two split peaks. (see  Figure 3)


    Figure 3. Peak-To-Valley ratio calculation


Figure 4. Integrating merged peaks

  • If cannot be avoided, you can use manual integration, see OpenLab CDS tutorial - Manually integrate a peak | Agilent

 Please note that in regulated environments  you always need to refer to the guidelines outlined in your SOPs for integration parameters.

 Tip  Learn how to effectively operate your OpenLab CDS:
HPLC-OLII-1140e - Agilent Infinity LC systems with OpenLab CDS - Integration and viewing results 
GC-OLII-2134eV2 - Basic Integration of GC Data Using OpenLab CDS Data Analysis (Ver. 2.4) e-learning courses available from Agilent education

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