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Nick Shulman responded: |
2018-04-25 15:53 |
Can you attach that file "exported_sml_molecule_translist.csv"?
I believe SkylineRunner expects the same sort of file as doing:
File > Import > Transition List
The first row of a small molecule transition list should be the column names ("PrecursorFormula", etc).
It's only if the first row does not have the column names that Skyline then tries to guess which column is the peptide sequence.
This is the complete list of column names that you can use in small molecule transition lists:
MoleculeGroup
PrecursorName
ProductName
PrecursorFormula
ProductFormula
PrecursorMz
ProductMz
PrecursorCharge
ProductCharge
PrecursorRT
PrecursorRTWindow
PrecursorCE
PrecursorDT
HighEnergyDTOffset
PrecursorIM
HighEnergyIMOffset
IMUnits
PrecursorCCS
SLens
ConeVoltage
CompensationVoltage
DeclusteringPotential
Note
LabelType
PrecursorAdduct
ProductAdduct
CAS
InChiKey
InChi
HMDB
SMILES
You can also put a space in the middle of some of those names, so that they look the column names that you see when you do:
Edit > Insert > Transition List |
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dspiciarich responded: |
2018-04-25 17:22 |
Maybe I'm misunderstanding but what I'm trying to do is look for peptides that won't be predicted by the peptide sequence so I'm trying to use the small molecule feature. I can do this in the gui but when try to do this with skylinerunner it is looking for peptides.
Also when I export my transition list and then try to reimport my transition list it throws an error that my transitions are for peptides. Any thoughts on how to resolve this?
Thanks,
David |
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Nick Shulman responded: |
2018-04-25 18:15 |
When you import a transition list into Skyline, you have to use column names that Skyline recognizes.
I can rename/remove some of the columns in the sml_translist.csv that you attached, and then Skyline is willing to import it:
MoleculeGroup,PrecursorName,PrecursorMz,PrecursorCharge,ProductMz,ProductCharge,PrecursorCE,ConeVoltage,ProductName
Compound 1,Molecule [2240.401097/2240.401097].light,1120.2,2,366.2,1,35.7,4,Ion [366.200549/366.200549]
Compound 2,Molecule [2345.401097/2345.401097].light,1172.7,2,366.2,1,37.4,4,Ion [366.200549/366.200549]
Compound 3,Molecule [2491.601097/2491.601097].light,1245.8,2,366.2,1,39.6,4,Ion [366.200549/366.200549]
If you you want to change all of the peptides in a Skyline document into small molecules, you can do a pretty good job by exporting a report that looks like a transition list, and then importing that.
Unfortunately, the report that comes with Skyline called "Small Molecule Transition List" does not have quite the right columns to be able to do this.
I have attached a .skyr file which does have a report that works.
You can get that .skyr file into Skyline by doing:
File > Export > Report > Edit List > Import
and then if you have a peptide document, you can export the report called "PeptideSmMolTranList" and you will have a .csv file that you can import into Skyline and get a document that measures the same transitions, but that treats everything as a small molecule.
I am still not sure that I understand your question, but I hope this helps. |
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dspiciarich responded: |
2018-04-25 18:21 |
Thank you Nick, I really appreciate it.
-David |
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warham responded: |
2025-08-07 11:25 |
Hi Nick,
Importing this molecule transition list works in the standard skyline interface (in any mode protein, mixed, or molecule) but does not work in the skylinerunner command line interface. Based on this discussion it looks like that skylinerunner looks for a precursor ion formula or precursor formula but that is no longer even an option (as seen in the drop down, captured, attached). The molecular formula and the precursor ion formula are of course different but it seems now the transition list import interface in the standard skyline gui supports only the molecular formula and then calculates the precursor formula from the charge/adduct info. If I have all that right, what is the convention for importing transition lists using skylinerunner command line, and is it possible to align the to two interfaces? |
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Nick Shulman responded: |
2025-08-07 11:47 |
I think you should call the column "MolecularFormula".
I actually thought "PrecursorFormula" would have worked as well. Somewhere in the Skyline source code there is a list of all of the column names which would be treated as synonyms for the "MolecularFormula" column, but I am not sure where that code lives.
For small molecules, if you haven't specified anything for the "Adduct" column, and all you have specified is the molecule formula and the charge, the molecule formula and the precursor formula are the same except for the number of electrons.
If you have specified an adduct, such as "[M+Na]", then the sodium gets added to the molecule formula to get the precursor ion formula. I think we thought it would be strange to specify a precursor formula that already included the sodium, so that Skyline would have to subtract the adduct to get back to the molecule formula.
I am not the expert on small molecules so I might be mistaken about some of this.
-- Nick |
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warham responded: |
2025-08-07 12:06 |
My column is called MolecularFormula, I also tried Molecular Formula. They both work in skyline gui, and I'm not able to get either to work in skylinerunner.
Thanks for responding so quickly. |
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Nick Shulman responded: |
2025-08-07 13:03 |
warham,
Can you send us your CSV file?
-- Nick |
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warham responded: |
2025-08-07 13:14 |
Here's a small one. Thank you |
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Nick Shulman responded: |
2025-08-07 14:35 |
warham,
What output are you seeing when you try to import that transition list using the command line interface?
I am not seeing any errors about column names, but I am seeing things like:
Error: (line 416, column 4) The precursor m/z 1685.805078 is not measureable with your current instrument settings.
Normally, I would be able to deal with that sort of error by saving a Skyline document where the instrument settings specify that the max mz is 2000.
However, when I specify "--in=" on the command line to tell Skyline to load a document with the correct settings, Skyline seems to be trying to import the transition list into a document with the wrong settings.
It sounds like you are running into a completely different issue than I am.
After I see the text of the output that you are seeing I will probably have a better idea of what is going wrong for you.
-- Nick |
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warham responded: |
2025-08-07 14:59 |
Hi Nick,
I have no errors with this file importing into skyline.
attached is the error I see with skyline runner. |
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warham responded: |
2025-08-07 15:02 |
that's a different tl file but the error is the same, sorry. |
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warham responded: |
2025-08-07 15:11 |
here's the same error from that file |
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Nick Shulman responded: |
2025-08-07 15:21 |
The error that you are seeing is:
Use --in to specify a Skyline document to open
The Skyline command line does require that you either user the "--in=" argument to tell Skyline to read a particular file, or "--new=" argument to tell Skyline to create a new document with the default settings.
I was able to successfully import your transition list, but I had to use the "--batch-commands" argument to specify a file with several lines of commands.
This is necessary because if you try to do it all in a single command line, Skyline will try to import the transition list into the current document before Skyline has opened the document that you specified with the "--in=" argument, so the settings will be wrong.
I got your transition list to work using the attached files, and the command line:
skylinerunner --batch-commands=commands.txt
The commands.txt file that I attached has the following lines in it:
--in=blank.sky
--import-transition-list=test_tl.csv
--out=foo.sky
This tells Skyline to open the file "blank.sky", then import the transition list and then save it as "foo.sky".
-- Nick
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warham responded: |
2025-08-14 15:22 |
so when I import the attached transition list into skyline by hand skyline calculates the precursor isotopic envelope ions. (The M0 is in the file and skyline calculates the M1-Mn, as usual).
When I import this transition list using skylinerunner it does not calculate the precursor isotopic envelope correctly, instead it generates the festival of excitement in the attached image. Also attached is the skylinerunner batch command file.
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Nick Shulman responded: |
2025-08-14 15:42 |
warham,
Can you send me the Skyline document that you ended up with?
You can open it up in Skyline and use the "File > Share" menu item to create a .zip file containing the Skyline document and supporting files.
The Share Document dialog gives you the option to include the .raw files, which would probably be helpful.
Files which are less than 50MB can be attached to these support requests.
You can always upload larger files here:
https://skyline.ms/files.url
I probably will also need that file "AcAnd_interior_ion_blank.sky" that you specified with the "--in" parameter in your commands.txt.
-- Nick |
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warham responded: |
2025-08-14 16:43 |
uploaded the zip file. data files are in folders (waters), the upload site doesn't support folders, not sure which files you need. |
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Nick Shulman responded: |
2025-08-14 17:01 |
warham,
One way to get Skyline to add the precursor isotope envelope transitions to the document is to open the document in the Skyline user interface and then use the menu item:
Refine > Advanced
In the Refine dialog, you can check the checkbox next to "Auto-select all...Transitions".
Because the "Ion types" textbox on the "Filter" tab at "Settings > Transition Settings" includes "p", Skyline will add precursor transitions to the document based on the "Isotope peaks included" on the "Full Scan" tab.
The way to do this on the command-line would be to have a line in that commands.txt which includes:
--refine-auto-select-transitions
-- Nick |
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