How to Match the Signal-To-Noise Calculations Between MassHunter Qualitative and Quantitative Analysis Software for Total Ion Chromatograms

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

This Information Applies To: All versions of Agilent MassHunter Qualitative Analysis Software , and Agilent MassHunter Quantitative Analysis version B08.00 and up

Issue

Calculation of signal-to-noise in MassHunter is performed using the same parameters for both Qualitative and Quantitative Analysis, however the parameters are entered with different steps. To match the results for both programs, the user must make sure to input the same parameters. The following procedure shows how to calculate the signal-to-noise and match the value in the Qualitative and Quantitative software for total ion chromatograms.

Steps to follow

Note: The following example uses MassHunter Qualitative Analysis version 10.0 and MassHunter Quantitative Analysis version 10.0. It applies the evaldemo.d data file (path X:\MassHunter\QuantExamples\MS\EVALDEMO\EVALDEMO.D) using the chromatographic peak at 6.43 minutes for biphenyl from the Eval A 10 ng/μL Standard.

In Qualitative Analysis, follow these steps:

Load the data file (File > Open Data File), select the evaldemo.d data file, then click Open, which will extract the TIC as the default chromatogram.
Go to View > Method Editor to display the Method Editor window.
1. In the Chromatogram drop down menu, click Integrate (MS) and select the appropriate integrator based on your application. The default integrator is Agile 2.
2. In the Chromatogram drop down menu, click Calculate Signal-to-Noise and select or enter the appropriate settings:
  
  Signal definition > Height
  
  Noise definition > Peak-to-Peak
  
  Specific noise regions > 6.600-7.400 (for Qualitative Analysis, use a hyphen between the noise region times).
Click Calculate Signal-to-Noise. This action will integrate the extracted ion chromatogram.

The signal-to-noise calculated is displayed in the Chromatogram Results window and Integration results table (View > Integration Peak List).

In Quantitative Analysis, follow these steps:

Create a new batch for the data file EVALDEMO.D in the folder X:\MassHunter\QuantExamples\MS\EVALDEMO. Click File> New batch, enter the batch name (Training), click Create, and then at the Add Samples window select the EVALDEMO.D data file.
Set the Type to Cal and Level to L1 in the batch table.
Click Method > New > New Method using Manual Setup to create the new Quantitative method for the batch.

From the Method Tasks window click the following Method Setup Tasks:
1. Compound Setup - fill in the Quantifier table row:
  
  Name > Biphenyl, TS > 1, Scan > Scan, Type > Target, MZ > 0, RT > 6.431, Ion Polarity > Positive, Criteria > Close RT.
  
  Tip: For a TIC signal, the MZ value must be set as zero.
2. Retention Time Setup - set both the Left RT Delta and Right RT Delta to 5.000 minutes so there is enough baseline signal available for the noise band calculation.
3. Concentration Setup - set Conc. to 10.000 and Units to ng/μL.
From the Method Tasks window click Advanced Tasks then click the following:
1. Integration Parameters Setup - select the Agile2 integrator in the Int. field of the Quantifier table.
2. Signal to Noise Setup - set Noise Regions to 6.600 7.400 (for Quant use a space between the noise regions times), Noise Alg. to Peak-to-Peak, and Noise SD Multiplier to 1.0 in the Quantifier table.
From the Method Tasks window click Save / Exit > Validate to check for any setup errors.

If there are no errors, or all errors have been fixed, click Save / Exit > Exit and select Analyze. Click Yes to apply the method to the batch and get the quantitative result.
The signal-to-noise value will be shown in the Batch Table. If the column is not displayed, right-click Biphenyl Results > Add Column > S/N.

It will match the value from the qualitative analysis result.