System Suitability
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System Suitability

In System Suitability you set the EIC and UV integration of QC standards.

This is necessary to correctly determine the UV Purity, MS and UV calibration curves, RT window, and other System Suitability calculations.

Page 1: EIC Integration

The List of Ions table shows all entries from the ion list with the type Ion. EEI and LEI ions have a corresponding flag in their entries.

The EIC Integration window shows the EIC of the selected ion in the first QC standard. The largest integrated peak, which will be used for calculations, is highlighted.

If necessary, adjust the peak start and end times manually as follows:

  1. In the List of Ions window, select the relevant ion.

  2. In the EIC Integration window, click and drag the integration points.

    Peak start and end points are updated in the table

  3. Click Integrate to apply potential retention time updates.

NOTE

Applying a changed EEI start time or a changed LEI end time will change the main peak start or end times on the UV Integration page. You will override manual corrections that you already applied on the UV Integration page.

EICs that have been manually integrated will be labeled with EIC manually integrated in the oligo analysis results report.

To check EIC integration of a different ion, select the corresponding entry in the table.
To compare multiple ions in an overlaid mode, hold the Ctrl key while selecting the ions.

Page 2: UV Integration

The QC Checks window lists all 25 µL Working Standard Solutions (QC standards) with their recommended peak start and end times, and resulting impurities. The UV Integration window shows the UV signal for the selected QC standard.

OAA recommends the UV integration as follows, based on EICs of early and late eluting impurities:

  • To determine the UV main peak start time, the wizard selects the earliest eluting known impurity (from the m/z ion list).

    Using the EIC for the earliest eluting known impurity, the software selects the integration time start, then subtracts the difference in retention times between FLP EIC and UV main peak.

    The UV main peak is split using this retention time as the new peak start time.

  • To determine the UV main peak end time, the wizard selects the latest eluting known impurity (from the m/z ion list).

    Using the EIC for the latest eluting known impurity, the software selects the integration time end, then subtracts the difference in retention times between FLP EIC and UV main peak.

    The UV main peak is split using this retention time as the new peak end time.

NOTE

To ensure proper integration, it is imperative that the UV main peak baseline has been established in the processing method.

If necessary, adjust the peak start and end times manually as follows:

  1. In the QC Checks window, select your first QC standard.

  2. In the UV Integration window, click and drag the dashed lines. Alternatively, enter the times in the input fields below the graph.

  3. Click Integrate to apply the modified settings to all QC standards.

Page 3: Results

The Results window provides an overview of the system suitability passing criteria.

Calibration curves are provided for both UV and MS signals.

UV plots the amount (in µg) of the pure drug substance relative to mAU. This mass is calculated based on injection volume (µL) and concentration (mg/mL), which is ultimately calculated based on the overall sample purity. The response is expected to be linear. The linear regression formula (y=ax+b) is displayed in a table below the Pass/Fail results.

MS plots injection volume (in µL) relative to the sum of FLP and FLP, P=O EIC areas. The response is expected to be correlated by second order polynomial regression, with the zero forced into the origin. The polynomial regression formula (y=Ax² + Bx) is displayed in a table below the Pass/Fail results.

All system suitability results are displayed and indicate pass or fail.

If all results passed, click Next to proceed. Confirm that you finished the system suitability settings. You will not be able to return to this step. Your changes are then saved as a new version of the result set.

  1. Based on the results, decide if you proceed with the Oligo analysis, or if you need to go back to OpenLab CDS Data Analysis.

  2. To proceed, click Next.

    If some of the tests failed, you will see a warning message. Confirm that you want to proceed with failed tests.

  3. Confirm that you finished the system suitability step. You will not be able to return to this step.

    Your changes are then saved as a new version of the result set.

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