How to manually create a calibration in Clarity Chromatography Software

Accurate calibration is a fundamental requirement for quantitative chromatography. Clarity Chromatography Software allows users to build and edit calibration curves from measured standards—even retroactively. This guide provides a detailed, step-by-step procedure for manually creating a calibration from already measured standards using the Calibration window.

The workflow described below is suitable for laboratories performing GC, HPLC, or LC/MS analysis where calibration data must be generated after the chromatographic runs have been completed.

Materials and Software Requirements

Software

• Clarity Chromatography Software (any edition supporting quantitative calibration)

Prerequisites

• Chromatograms of measured calibration standards
• Concentrations or amounts of each compound in every standard level
• An empty calibration file created or opened in Clarity

Overview of the Calibration Workflow

1. Open an empty calibration file
2. Load the first standard
3. Import integrated peaks into the calibration table
4. Name compounds and enter concentrations
5. Load the next standard level
6. Add only existing compounds to the next level
7. Enter concentrations for all remaining calibration levels
8. Link the calibration to the chromatogram and verify calculated results

Step-by-Step Procedure

1. Open or Create an Empty Calibration File

Begin by opening the Calibration window in Clarity.

• Either create a new calibration file
• Or open a previously prepared empty file

This ensures all following actions populate a clean calibration structure.

2. Load the First Standard Chromatogram

Open the chromatogram corresponding to the first calibration level.

3. Add All Integrated Peaks

Use the Add All function to import all integrated peaks into the calibration.

• All detected peaks are transferred into the Calibration Summary Table.
• This establishes the initial list of compounds for the calibration.

Important:
Calibration compound names are identical across all levels and must be set only once.

4. Rename Compounds (If Needed)

In the Calibration Summary Table:

• Rename each peak to its correct compound name.
• Ensure consistency, as these names will be used for all subsequent levels.

5. Enter Compound Amounts for Level 1

For every compound:

• Enter the appropriate amount, concentration, or mass corresponding to the first calibration level.

This completes Level 1 of the calibration.

6. Load the Second Standard Chromatogram

Open the chromatogram corresponding to the second calibration level.

Verify Toolbar Settings

Before adding peaks, ensure the toolbar settings are correct:

• Level:2
• Mode:Automatic
• Calibration:Enabled

Correct settings ensure values are added to the appropriate calibration level.

7. Add Existing Compounds Only

Instead of adding all peaks, use the Add Existing function:

• This imports only the compounds already defined in Level 1.
• Helps maintain consistent compound identity across all levels.

The Calibration Summary Table automatically switches to Level 2.

8. Enter Compound Amounts for Level 2

Just as with Level 1:

• Fill in the required amounts for each compound for the second calibration point.

Repeat Steps 6–8 for every additional calibration level.

9. Verify Calibration in the Chromatogram Window

Switch to the Chromatogram window where the calibration is linked.

You should now see:

• Correct compound names displayed above peaks
• Quantitative results automatically calculated
• The calibration curve applied to the chromatogram

This confirms that the manually constructed calibration is active and functioning.

Discussion

The manual calibration procedure is useful when:

• Standards were run before a calibration file was prepared
• Analysts need to reconstruct or verify quantitation
• Retrospective calibration is necessary after data review

Advantages of manual calibration building:

• Full control over compound naming
• Ability to exclude unwanted peaks
• Exact control over concentration entries
• Flexible creation of multi-level calibrations

Conclusion

Creating a manual calibration from existing chromatographic standards in Clarity is a straightforward process when following a structured workflow. By using functions such as Add All and Add Existing, assigning accurate compound names, and entering correct standard amounts for each level, users can efficiently generate precise calibration curves for quantitative analysis.

This manual calibration method is ideal for laboratories that need to reconstruct, edit, or refine calibration data outside of automated acquisition workflows.

For more instructional content and advanced Clarity tutorials, visit the official DataApex YouTube channel.