Xu Li-hua
Carlos VargasThis Information Applies To: Agilent MassHunter Qualitative and Quantitative Analysis
Issue
Calculation of signal-to-noise in MassHunter is done using the same routine for both Qualitative and Quantitative Analysis. However, there are differences in the way that the parameters are input. To get the same value between the programs, the user must make sure that they are inputting the same parameters. The following procedure is to calculate the signal-to-noise and get the same value in the Qualitative and Quantitative software for extraction chromatograms.
Steps to follow
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Note: The following example uses MassHunter Qualitative Analysis version 10.0 and MassHunter Quantitative Analysis version 10.0 with the evaldemo.d data file (path X:\MassHunter\QuantExamples\MS\EVALDEMO\EVALDEMO.D). This example uses the chromatographic peak at 6.43 minutes for Biphenyl from the Eval A 10 ng/μL Standard. |
In Qualitative Analysis, follow the steps below:
1. Load data evaldemo.d and extract chromatogram by clicking the menu Chromatograms > Extract Chromatograms(see Figure 1).
Figure 1. Extract chromatograms
2. Input the Extracted Ion Chromatogram parameters. In the MS Chromatogram tab, choose the Type EIC and set m/z value 154.1. Switch to the Advanced tab, set the Single m/z Expansion for this chromatogram to Symmetric (m/z) and ± 0.5, which will match the value set in the MassHunter Quantitative Analysis software, then press OK (see Figure 2).
Figure 2. Setup parameters of extract chromatograms
a. Select the appropriate integrator based on your application (see Figure 3), the default integrator is Agile 2 from the Chromatograms > Integrate (MS) > Integrator tab.
b. Set the signal-to-noise calculating parameters (see Figure 4).
c. Set the Signal definition, Noise measurement, and Specific noise regions from the Chromatograms > Calculate Signal-to-Noise > Signal Measurement tab.
d. Set Signal definition: to Height, Noise definition: to Peak-to-Peak and Specific noise regions to 6.600–7.400 min (use a hyphen between the Noise Region Times).
Figure 4. Calculate Signal-to-Noise option
4. Click the icon Calculate Signal-to-Noise (see Figure 5) – this will Integrate the Extracted Ion Chromatogram and display the calculated Signal-to-Noise result in the Integration Peak List (if it isn’t displayed use View > Integration Peak List).
Figure 5. Calculate Signal-to-Noise action
5. The signal-to-noise calculated result is in Figure 6.
Figure 6. Signal-to-noise calculated result
In Quantitative Analysis, follow the steps below:
6. Create a new batch for the data file EVALDEMO.D in the folder X:\MassHunter\QuantExamples\MS\EVALDEMO. To do this, click File> New batch, enter the batch name, click Create, and choose the EVALDEMO.D data file, see Figure 7).
7. Set the Type to Cal and Level to L1 in the batch table (see Figure 8).
8. Edit the Quantitative Method – Method > Edit Method Tasks > Manual Setup Tasks > New Compound creates a new compound. Then fill in the row in Compound Setup > Name Biphenyl, TS 1, Scan Scan, Type Target, MZ 154.1, RT 6.431, Ion Polarity Positive, Criteria Close RT (see Figure 9).
Change both the Left RT Delta and Right RT Delta in Retention Time Setup to 1.500 Minutes so there is available signal for the noise band calculation.
Create a calibration level from menu Method > Create Levels from Calibration Samples. Enter the concentration in the Concentration Setup as 10.000 and change the Units to ng/μL.
Method Tasks > Advanced Tasks > Integration Parameters Setup choose the Agile2 integrator.
Method Tasks > Advanced Tasks > Signal to Noise Setup set the Noise Regions 6.600 7.400, Noise Alg. to Peak-to-Peak, Noise SD Multiplier to 1.0 (Note: Use a space between the Noise Region Times).
Method Tasks > Advanced Tasks > Mass Extraction Setup set the Quantifier Extract Left m/z and Extract Right m/z to 0.50.
See Figure 10.
9. Method Tasks > Save / Exit > Validate to check for any setup errors. If no errors are found, or once errors have been rectified then Method Tasks > Save / Exit > Exit, then select Analyze, and click Yes to get the quantitative result (see Figure 11).
10. The signal-to-noise value shown in the batch table will be the same as the qualitative result (see Figure 12). (Note: Right click Biphenyl Results > Add Column > S/N if the column isn’t displayed).
Figure 12. Signal-to-noise calculated result
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Learn more on how to effectively operate your MassHunter software, please refer to the following course:
Agilent 5977 GC/MSD with MassHunter Workstation e-learning path available from Agilent education |
