Dear all, another query r.e. Waters QTOF data....

We have a QTOF Premier which is now being used for SILAC experiments,
and we want to quantify using ASAPRatio / Xpress. Have no problems
doing this in our pipeline using Orbitrap data, but the QTOF data is
causing headaches. Up until now we've mostly been using MassWolf just
to get MS data into .mzXML for purposes other than ID & quant, PLGS
having been the preferred analysis tool for the instrument.

If we use .mzXML files and feed them to Mascot/OMSSA/Tandem + TPP we
cannot get anywhere near the number of IDs as with .pkl files
generated using PLGS. We've tried conversion to mzXML using MassWolf
and msconvert, applying lockmass correction in MassLynx prior to
conversion, and comparing acquisition of MS/MS scans in centroid mode,
vs acquisition in profile mode converted to centroid using MassLynx.
An example of the differences in IDs:

RAW->PLGS->PKL->Mascot->TPP: 874 peptide IDs (1% FDR)
RAW->MassWolf->mzXML->mgf->Mascot->TPP: 279 peptide IDs (1 % FDR)

No matter what combination of converter / pre-processing in MassLynx
we try, the number of IDs is never above 1/3 of that we get from a PKL
file. We've also tried changes to the acquisition methods, without
success.

Is anyone able to provide a MassLynx method file, and/or procedure for
pre-processing / .mzXML conversion they are using successfully with a
QTOF Premier or similar instrument?

Any help would be gratefully appreciated. Many Thanks,

DT
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