How does SkylineCmd.exe calculate chromatographic area?

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How does SkylineCmd.exe calculate chromatographic area? lparsons  2017-09-21 08:11
 
If I import raw data into SkylineCmd.exe and use the --chromatogram-file option to export chromatograms to a tab delimited file, how are the peak areas calculated?

I had - perhaps erroneously - expected that the peak area would be the sum of the intensities for the ion in question. I am using a transition list that is already in the template file, which is looking only at MS1 data. However I found that if I take the intensities from the exported chromatogram file and sum up the intensities of just the window that is +/- 60 seconds of the most intense peak for the ion m/z in question, I often get a sum that far exceeds the area reported for the chromatogram.

Is the area calculated differently? I may have missed a tutorial that better describes how this is done; please feel free to refer me to the one I should review for this if so.

Thank you
Lee
 
 
Nick Shulman responded:  2017-09-21 14:38
When you extract chromatograms using SkylineCmd.exe, you should end up getting the same numbers as you would have gotten if you had done the same work through the regular Skyline user interface.

One difference between what you get by just summing chromatogram intensities, and what Skyline computes might involve background subtraction. Here is a page that talks about how Skyline calculates background by drawing a horizontal line across the peak at the lower of either the start or end heights of the peak:
https://skyline.ms/wiki/home/software/Skyline/page.view?name=tip_peak_calc