Retention time difference in library when using PDresult vs. msf file

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Retention time difference in library when using PDresult vs. msf file roberthardt  2019-04-23 07:38
 

Dear Skyline-Team,

I today created a spectral library from ProteomeDiscoverer 2.3 results (Mascot search engine) once using the .PDresult and once using the .msf file(s). I got quite similar results but interestingly, there is a clear retention time shift/offset when using the PDresult file. This becomes immediately apparent when looking at MS1-chromatograms, where the blue vertical peptide-ID lines are clearly "missing" the correct MS1-peak in the case of the PDresults-library.

Best

Robert

 
 
Nick Shulman responded:  2019-04-23 12:41
Can you send us your .pdresult file and your .msf file? I am not sure whether there are other files we need when building a library from these types of files. I imagine we will also need to see that .raw file in order for us to understand what is going wrong.

Files that are less than 50MB can be attached to this support request. You can upload larger files here:
https://skyline.ms/files.url

-- Nick
 
roberthardt responded:  2019-04-24 01:54
Hi Nick,

I will upload the files in a single zip-file named 190424_roberthardt.zip

Best

Robert
 
Matt Chambers responded:  2019-04-25 12:16

Hi Robert,

I looked at your MSF and pdResult files and the problem seems to lie in your workflow. The retention times in the pdResult file have been modified. Possibly due to some alignment process. Here's a sampling to compare the times in the msf vs pdResult. You can see that the ratio starts out wildly different but as the file progresses it approaches 1. Strangely, for the 19_PRM file, the ratio starts out at 1 and then has a minor adjustment sometime later, but as the file progresses the ratio falls back toward 1. In other words, I have no idea what's going on, but it's not something we can fix in BiblioSpec. :)

RawFile ScanNumber PdResultTime MsfTime TimeRatio
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190405_P_315_WV_018_HCD.raw 5 4.15688372736299 0.0141472456 293.82989769846
190405_P_315_WV_018_HCD.raw 6 4.16135287562966 0.0186163938666667 223.531630531341
190405_P_315_WV_018_HCD.raw 8 4.16571273536299 0.0229762536 181.305133895414
190405_P_315_WV_018_HCD.raw 12 4.18629161242966 0.0435551306666667 96.114775649921
190405_P_315_WV_018_HCD.raw 20 4.23024967294633 0.0875131911833333 48.3384232222121
190405_P_315_WV_018_HCD.raw 22 4.23668515347966 0.0939486717166667 45.0957429846032
190405_P_315_WV_018_HCD.raw 28 4.26893898227966 0.126202500516667 33.8261045922453
190405_P_315_WV_018_HCD.raw 34 4.30015769722966 0.157421215466667 27.3162526695152
190405_P_315_WV_018_HCD.raw 36 4.30636783882966 0.163631357066667 26.3174975507608
190405_P_315_WV_018_HCD.raw 41 4.33342400949633 0.190687527733333 22.7252619036332
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