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      • Creating a New Batch in MassHunter Quantitative Analysis
      • Creating a Method in MassHunter Quantitative Analysis – Quant-My-Way user interface
      • Calculating the Final Concentration in MassHunter Quantitative Analysis
      • How to Average Calibration Responses in MassHunter Quantitative Analysis
      • Outlier Setup in MassHunter Quantitative Analysis Software
      • Combine multiple Methods in MassHunter Quantitative Analysis into a Single Method
      • Using the Compound Information window in MassHunter Quantitative Analysis Software
      • Modify the Integration Parameters in MassHunter Quantitative Analysis
      • Using the Batch Table in MassHunter Quantitative Analysis Software
      • Using the Calibration Curve Window in MassHunter Quantitative Analysis Software
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How to Average Calibration Responses in MassHunter Quantitative Analysis

Created by Xu Li-hua Xu Li-hua over 1 year ago | Last modified by Carlos Vargas Carlos Vargas over 1 year ago

This Information Applies To: Agilent MassHunter Quantitative Analysis Software


Issue

When creating a MassHunter Quantitative Analysis Batch with many samples, several sets of calibration standards may be included. If your protocol requires the use of the average response from several sets of calibration standards to define the calibration curve, this article demonstrates the steps needed to perform this operation in MassHunter Quantitative Analysis.


Steps to Follow

The steps below are demonstrated in MassHunter Quantitative Analysis version 10.0.

1. Initially create a calibration table based on all the sets of calibration standards for all concentration levels in the Batch (see Figure 1). 

Figure 1. View > Batch Table Layout > Flat Table

2. Click Analyze Batch to process the batch and populate the calibration table with the calibration responses. The Calibration Curve window will now show the number of Levels and Points currently in use. In this example, 7 Levels and 14 Points (two Points per Level) based on the multiple levels and points as defined in the Batch Table (see Figure 2).


Figure 2. The calibration curve window of 7 levels and 14 points

3. Edit the Method – Method > Edit or F10 (see Figure 3).


Figure 3. Method > Edit

4. Click Method Tasks > Method Setup Tasks > Concentration Setup. Verify for each compound that all the calibration sets levels and responses are populated in the Method Table (Figure 4).

 Tip 
Note: If required the concentration values (Conc.) can be reviewed and adjusted for situations where a compound does not have the same concentration as the value set for Expected Concentration (Exp. Conc.) in the Batch Table.


Figure 4. Concentration setup table

5. From the Method Edit view, click Method > Average Calibration Replicates (see Figure 5).


Figure 5. Average Calibration Replicates option

6. Select All or click the individual compounds to be calibration averaged (see Figure 6).

 Tip 
Note: If the quantitation method uses internal standards, then all the compounds associated with a given internal standard MUST be selected. 


Figure 6. Compounds selection dialogue box for average calibration

7. Review the Concentration Setup table, there will now be a single level per concentration instead of the previous multiple levels per concentration (see Figure 7).


Figure 7. Concentration setup table

8. Select Exit in the Method Setup Tasks and when Apply Method dialogue box is displayed, select Quantitate and then click Yes (see Figure 8).

 Tip 
Note: Do NOT select Analyze - Selecting Analyze will recreate a new calibration curve based on the sets of calibration standards and concentration levels defined in the Batch Table.


Figure 8. The Apply Method dialogue box

9. The Calibration Curve window will now show the number of Levels and Points currently in use after the Average Calibration Replicates step. In this example, there are now seven Levels and seven Points (one Point per Level) (see Figure 9).


Figure 9. One point per level in the calibration curve window after Average calibration replicates step

10. Only reprocess the batch using Quantitate Batch – avoid reprocessing using Analyze Batch as this will recreate a new calibration curve based on the sets of calibration standards and concentration levels defined in the Batch Table (see Figure 10). 

Quantitate Batch will retain the current Average Calibration Replicates… single level per concentration and process all samples using that protocol.


Figure 10. Quantitate batch option

 Tip
Learn how to effectively operate your Agilent MassHunter Software:
Agilent 5977 GC/MSD with MassHunter Workstation
Agilent 7000 or 7010 Triple Quadrupole GC/MS with MassHunter Workstation 
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