genform.xml

<tool id="genform" name="genform" version="@GENFORM_VERSION@" python_template_version="3.5">
    <description>Generation of molecular formulas by high-resolution MS and MS/MS data</description>
    <macros>
        <token name="@GENFORM_VERSION@">8</token>
    </macros>
    <requirements>
        <requirement type="package" version="r@GENFORM_VERSION@">genform</requirement>
    </requirements>
    <command detect_errors="exit_code"><![CDATA[
genform 
  ms='$ms'
  #if $msms
    msms='$msms'
  #end if
  #if $formula_cond.formula_select != 'advanced'
    $formula_cond.formula_select
  #else
    $formula_cond.vsp
    #if str($formula_cond.vsm2mv)
      vsm2mv=$formula_cond.vsm2mv
    #end if
    #if str($formula_cond.vsm2ap2)
      vsm2ap2=$formula_cond.vsm2ap2
    #end if
    #if str($formula_cond.ivsm2mv)
      ivsm2mv=$formula_cond.ivsm2mv
    #end if
    #if str($formula_cond.ivsm2ap2)
      ivsm2ap2=$formula_cond.ivsm2ap2
    #end if
  #end if
  #if str($m)
    m=$m
  #end if
  ion=$ion
  ppm=$ppm
  $msmv
  acc=$acc
  rej=$rej
  thms=$thms
  thmsms=$thmsms
  thcomb=$thcomb
  #if $sort
    $sort
  #end if	
  #if $elemfilt_cond.elemfilt_select == 'elem'
    el=#echo "".join(str($elemfilt_cond.el).split(','))
  #elif $elemfilt_cond.elemfilt_select == 'oc'
    oc
  #elif $elemfilt_cond.elemfilt_select == 'fuzzy'
    ff=$elemfilt_cond.ff
  #end if
  $het
  $hcf
  $wm
  $wi_cond.wi
  #if $wi_cond.wi == 'wi=log' and str($wi_cond.exp)
    exp=$wi_cond.exp
  #end if
  $oei
  #if str($dbeexc)
    dbeexc=$dbeexc
  #end if
  #if $output.out_cond.out_select == 'analyze':
      analyze
      $output.out_cond.loss
      $output.out_cond.intens
  #end if
  $output.dbe
  $output.cm
  $output.pc
  $output.sc
  out=out
  #if $addoutput.oms
     oms='$outoms'
  #end if
  #if $addoutput.omsms
     omsms='$outomsms'
  #end if
  #if $addoutput.oclean
     oclean='$outoclean'
  #end if

  ## for some reason genform prints the file twice if sorting is selected
  ## (once unsorted and once sorted). hence, if soring is enabled just
  ## the lower half of the file is returned
  #if $sort
  &&
    wc=\$(wc -l out | cut -d" " -f 1 )
  &&
    hwc=\$(echo "\$wc/2" | bc)
  &&
    tail -n \$hwc > '$out'
  #else
    && cp out '$out'
  #end if
    ]]></command>
    <inputs>
        <param type="data" argument="ms" label="MS data" format="tabular" help="" />
        <param type="data" argument="msms" label="MS/MS data (optional)" optional="true" format="tabular" help="" />
        <conditional name="formula_cond">
            <param type="select" name="formula_select" label="Structural formula existence check" help="Allow only molecular formulas for that at least one structural formula exists or specify advanced restictions, note that the default of genform is 'no existence check'">
                <option value="">No existence check</option>
                <option value="exist" selected="true">Only molecular formulas for that at least one structural formula exists (exist)</option>
                <option value="exist=mv">Enable multiple valencies for P and S (exist=mv)</option>
                <option value="advanced">Advanced</option>
             </param>
             <when value=""/>
             <when value="exist"/>
             <when value="exist=mv"/>
             <when value="advanced">
                <param type="select" argument="vsp" label="Valency sum parity" help="Even for graphical formulas" >
                    <option value="">Use default</option>
                    <option value="vsp=even">Even</option>
                    <option value="vsp=odd">Odd</option>
                </param>
                <param type="integer" argument="vsm2mv" label="Lower bound for valency sum - 2 * maximum valency for graphical formulas" min="0" value="" help="" />
                <param type="integer" argument="vsm2ap2" label="Lower bound for valency sum - 2 * number of atoms + 2 for graphical connected formulas" min="0" value="" help="" />
                <param type="integer" argument="ivsm2mv" label="Lower bound for valency sum - 2 * maximum valency for fragment ions" min="0" value="" help="" />
                <param type="integer" argument="ivsm2ap2" label="Lower bound for valency sum - 2 * number of atoms + 2 for fragment ions" min="0" value="" help="" />
             </when>
        </conditional>
        <param type="float" argument="m" label="Experimental molecular mass" min="0" optional="true" help="(default: mass of MS basepeak)" />
        <param type="select" argument="ion" label="Type of ion measured" help="" >
            <option value="M+H">[M+H]+</option>
            <option value="M-H">[M-H]-</option>
            <option value="M+e">M+</option>
            <option value="M-e">M-</option>
            <option value="M+Na">[M+Na]+</option>
        </param>
        <param type="integer" argument="ppm" label="Accuracy of measurement of MS1" min="0" value="5" help="In parts per million" />
        <param type="select" argument="msmv" label="MS match value" help="Select output type of the MS match value" >
            <option value="msmv=ndp">Normalized dot product</option>
            <option value="msmv=nsse">Normalized sum of squared</option>
            <option value="msmv=nsae" selected="true">Absolute errors</option>
        </param>
        <param type="integer" argument="acc" label="Allowed deviation for full acceptance" min="0" value="2" help="of MS/MS peak in ppm" />
        <param type="integer" argument="rej" label="Allowed deviation for total rejection" min="0" value="4" help="of MS/MS peak in ppm" />
        <param type="float" argument="thms" label="Threshold for the MS match value" min="0" max="1" value="0" help="" />
        <param type="float" argument="thmsms" label="Threshold for the MS/MS match value" min="0" max="1" value="0" help="" />
        <param type="float" argument="thcomb" label="Threshold for the combined match value" min="0" max="1" value="0" help="" />
        <param type="select" argument="sort" optional="true" label="Sort generated formulas by" help="" >
            <option value="sort=ppm">Mass deviation in ppm</option>
            <option value="sort=msmv">MS match value</option>
            <option value="sort=msmsmv">MS/MS match value</option>
            <option value="sort=combmv">Combined match value</option>
        </param>
        <conditional name="elemfilt_cond">
            <param type="select" name="elemfilt_select" label="Chemical element filtering">
                <option value="nofilter">No filter</option>
                <option value="elem">By used elements</option>
                <option value="oc">Only organic compounds (oc)</option>
                <option value="fuzzy">By fuzzy formula</option>
	    </param>
	    <when value="nofilter"/>
            <when value="elem">
                <param type="select" multiple="true" argument="el" label="Used chemical elements" help="" >
                    <option selected="true" value="C">C</option>
                    <option selected="true" value="N">N</option>
                    <option selected="true" value="O">O</option>
                    <option selected="true" value="P">P</option>
                    <option selected="true" value="S">S</option>
                    <option selected="true" value="H">H</option>
                    <option selected="true" value="Cl">Cl</option>
                    <option selected="true" value="Br">Br</option>
                    <option selected="true" value="F">F</option>
                    <option selected="true" value="I">I</option>
                    <option selected="true" value="Si">Si</option>
                </param>
             </when>
             <when value="oc"/>
             <when value="fuzzy">
                 <param type="text" argument="ff" checked="false" label="Uses fuzzy formula for limits of element multiplicities" help="" />
             </when>
        </conditional>
        <param type="boolean" argument="het" checked="false" truevalue="het" falsevalue="" label="Formulas must have at least one hetero atom" help="" />
        <param type="boolean" argument="hcf" checked="false" truevalue="hcf" falsevalue="" label="Apply Heuerding-Clerc filter" help="" />
        <param type="select" argument="wm" label="Weighting of m/z for MS/MS match value" help="" >
            <option value="">No weighting</option>
            <option value="wm=lin">Linear</option>
            <option value="wm=sqrt">Square root</option>
            <option value="wm=log">Logarithmic</option>
        </param>
        <conditional name="wi_cond">
            <param type="select" argument="wi" label="Intensity weighting for MS/MS match value" help="" >
                <option value="">No weighting</option>
                <option value="wi=lin">Linear</option>
                <option value="wi=sqrt">Square root</option>
                <option value="wi=log">Logarithmic</option>
            </param>
            <when value=""/>
            <when value="wi=lin"/>
            <when value="wi=sqrt"/>
            <when value="wi=log">
                <param type="float" argument="exp" optional="true" label="Exponent" value="" help="" />
            </when>
        </conditional>
        <param type="boolean" argument="oei" checked="false" truevalue="oei" falsevalue="" label="Allow odd electron ions for explaining MS/MS peaks" help="" />
        <param type="integer" argument="dbeexc" optional="true" label="Excess of double bond equivalent for ions" help="" />
        <section name="output" title="Output options for main output" expanded="true">
            <conditional name="out_cond">
                <param type="select" name="out_select" label="Write explanations for MS/MS peaks to output" help="" >
                    <option value="">no</option>
                    <option value="analyze">yes</option>
                </param>
                <when value=""/>
                <when value="analyze">
                    <param type="boolean" argument="loss" checked="false" truevalue="loss" falsevalue="" label="For analyzing MS/MS peaks write losses instead of fragments" help="" />
                    <param type="boolean" argument="intens" checked="false" truevalue="intens" falsevalue="" label="Write intensities of MS/MS peaks to output" help="" />
                </when>
            </conditional>
            <param type="boolean" argument="dbe" checked="false" truevalue="dbe" falsevalue="" label="Write double bond equivalents to output" help="" />
            <param type="boolean" argument="cm" checked="false" truevalue="cm" falsevalue="" label="Write calculated ion masses to output" help="" />
            <param type="boolean" argument="pc" checked="false" truevalue="pc" falsevalue="" label="Output match values in percent" help="" />
            <param type="boolean" argument="sc" checked="false" truevalue="sc" falsevalue="" label="Strip calculated isotope distributions" help="" />
        </section>
        <section name="addoutput" title="Additional output data sets" expanded="true">
            <param type="boolean" argument="oms" checked="false" truevalue="oms" falsevalue="" label="Write scaled MS peaks output" help="" />
            <param type="boolean" argument="omsms" checked="false" truevalue="omsms" falsevalue="" label="Write scaled MS/MS peaks output" help="" />
            <param type="boolean" argument="oclean" checked="false" truevalue="oclean" falsevalue="" label="Write MS/MS with only explained peaks" help="" />
            <param type="boolean" argument="oclean" checked="false" truevalue="oms" falsevalue="" label="Write MS/MS with only explained peaks" help="" />
        </section>
    </inputs>
    <outputs>
        <data name="out" format="tabular" />
        <data name="outoms" format="tabular" label="${tool.name} on ${on_string}: MS">
            <filter>addoutput['oms']</filter>
        </data>
        <data name="outomsms" format="tabular" label="${tool.name} on ${on_string}: MS/MS" >
            <filter>addoutput['omsms']</filter>
        </data>
        <data name="outoclean" format="tabular" label="${tool.name} on ${on_string}: clean">
            <filter>addoutput['oclean']</filter>
        </data>
    </outputs>
    <tests>
        <!-- test with MS input and mostly default values -->
        <test expect_num_outputs="1">
            <param name="ms" ftype="tabular" value="CreatineMs.txt" />
            <param name="acc" value="20" />
            <param name="rej" value="40" />
            <output name="out" ftype="tabular" file="CreatineMs.out"/>
            <assert_command>
                <has_text text=" ms=" />
                <not_has_text text=" msms=" />
                <has_text text=" exist " />
                <not_has_text text=" m=" />
                <has_text text=" ion=M+H " />
                <has_text text=" ppm=5 " />
                <has_text text=" msmv=nsae " />
                <has_text text=" acc=20 " />
                <has_text text=" rej=40 " />
                <has_text text=" thmsms=0.0 " />
                <has_text text=" thmsms=0.0 " />
                <has_text text=" thcomb=0.0 " />
                <not_has_text text=" sort" />
                <not_has_text text=" el=" />
                <not_has_text text=" oc " />
                <not_has_text text=" ff=" />
                <not_has_text text=" het " />
                <not_has_text text=" vsp=" />
                <not_has_text text=" vsp2mv=" />
                <not_has_text text=" vsm2ap2=" />
                <not_has_text text=" hcf " />
                <not_has_text text=" wm=" />
                <not_has_text text=" wi=" />
                <not_has_text text=" exp=" />
                <not_has_text text=" oei " />
                <not_has_text text=" dbeexc=" />
                <not_has_text text=" ivsm2mv=" />
                <not_has_text text=" ivsm2ap2=" />
                <not_has_text text=" oms=" />
                <not_has_text text=" omsms=" />
                <not_has_text text=" oclean=" />
                <not_has_text text=" analyze " />
                <not_has_text text=" loss " />
                <not_has_text text=" intens " />
                <not_has_text text=" dbe " />
                <not_has_text text=" cm " />
                <not_has_text text=" pc " />
                <not_has_text text=" sc " />
            </assert_command>
        </test>
        <!-- test with MS/MS input and non default output options -->
        <test expect_num_outputs="4">
            <param name="ms" ftype="tabular" value="CreatineMs.txt" />
            <param name="msms" ftype="tabular" value="CreatineMsMs.txt" />
            <param name="acc" value="20" />
            <param name="rej" value="40" />
            <section name="output">
                <conditional name="out_cond">
                    <param name="out_select" value="analyze"/>
                    <param name="loss" value="loss" />
                    <param name="intens" value="intens" />
                </conditional>
                <param name="dbe" value="dbe" />
                <param name="cm" value="cm" />
                <param name="pc" value="pc" />
                <param name="sc" value="sc" />
            </section>
            <section name="addoutput">
                <param name="oms" value="oms" />
                <param name="omsms" value="omsms" />
                <param name="oclean" value="oclean" />
                <param name="oclean" value="oms"/>
            </section>
            <output name="out" ftype="tabular" file="CreatineMsMs.out"/>
            <output name="outoms" ftype="tabular" file="CreatineMsMs-oms.out"/>
            <output name="outomsms" ftype="tabular" file="CreatineMsMs-omsms.out"/>
            <output name="outoclean" ftype="tabular" file="CreatineMsMs-oclean.out"/>
            <assert_command>
                <has_text text=" ms=" />
                <has_text text=" msms=" />
                <has_text text=" exist" />
                <not_has_text text=" m=" />
                <has_text text=" ion=M+H " />
                <has_text text=" ppm=5 " />
                <has_text text=" msmv=nsae " />
                <has_text text=" acc=20 " />
                <has_text text=" rej=40 " />
                <has_text text=" thmsms=0.0 " />
                <has_text text=" thmsms=0.0 " />
                <has_text text=" thcomb=0.0 " />
                <not_has_text text=" sort" />
                <not_has_text text=" el=" />
                <not_has_text text=" oc " />
                <not_has_text text=" ff=" />
                <not_has_text text=" het " />
                <not_has_text text=" vsp=" />
                <not_has_text text=" vsp2mv=" />
                <not_has_text text=" vsm2ap2=" />
                <not_has_text text=" hcf " />
                <not_has_text text=" wm=" />
                <not_has_text text=" wi=" />
                <not_has_text text=" exp=" />
                <not_has_text text=" oei " />
                <not_has_text text=" dbeexc=" />
                <not_has_text text=" ivsm2mv=" />
                <not_has_text text=" ivsm2ap2=" />
                <has_text text=" oms=" />
                <has_text text=" omsms=" />
                <has_text text=" oclean=" />
                <has_text text=" analyze " />
                <has_text text=" loss " />
                <has_text text=" intens " />
                <has_text text=" dbe " />
                <has_text text=" cm " />
                <has_text text=" pc " />
                <has_text text=" sc " />
            </assert_command>
        </test>
        <!-- test with MS/MS input and other non-default values 
             (creates empty output .. but test is still usefull) -->
        <test expect_num_outputs="1">
            <param name="ms" ftype="tabular" value="CreatineMs.txt" />
            <param name="msms" ftype="tabular" value="CreatineMsMs.txt" />
            <conditional name="formula_cond">
                <param name="formula_select" value=""/>
            </conditional>
            <param name="m" value="111.0201" />
            <param name="ion" value="M-H" />
            <param name="ppm" value="4" />
            <param name="msmv" value="msmv=ndp"/>
            <param name="acc" value="21" />
            <param name="rej" value="39" />
            <param name="thms" value="0.3" />
            <param name="thmsms" value="0.2" />
            <param name="thcomb" value="0.1" />
            <param name="sort" value="sort=combmv" />
            <conditional name="elemfilt_cond">
                <param name="elemfilt_select" value="elem"/>
                <param name="el" value="C,N,O,P,S,H,Cl,Br,F,I" />
            </conditional>
            <param name="het" value="het" />
            <param name="hcf" value="hcf" />
            <param name="wm" value="wm=sqrt"/>
            <conditional name="wi_cond">
                <param name="wi" value="wi=log"/>
                <param name="exp" value="2.31"/>
            </conditional>
            <param name="oei" value="oei" />
            <param name="dbeexc" value="1" />
            <output name="out" ftype="tabular" file="CreatineMsMs.out" lines_diff="3"/>
            <assert_command>
                <has_text text=" ms=" />
                <has_text text=" msms=" />
                <not_has_text text=" exist" />
                <has_text text=" m=111.0201" />
                <has_text text=" ion=M-H" />
                <has_text text=" ppm=4" />
                <has_text text=" msmv=ndp" />
                <has_text text=" acc=21" />
                <has_text text=" rej=39" />
                <has_text text=" thms=0.3" />
                <has_text text=" thmsms=0.2" />
                <has_text text=" thcomb=0.1" />
                <has_text text=" sort=combmv" />
                <has_text text=" el=CNOPSHClBrFI" />
                <not_has_text text=" oc " />
                <not_has_text text=" ff=" />
                <has_text text=" het " />
                <not_has_text text=" vsp=" />
                <not_has_text text=" vsp2mv=" />
                <not_has_text text=" vsm2ap2=" />
                <has_text text=" hcf " />
                <has_text text=" wm=sqrt " />
                <has_text text=" wi=log " />
                <has_text text=" exp=2.31 " />
                <has_text text=" oei " />
                <has_text text=" dbeexc=1" />
                <not_has_text text=" ivsm2mv=" />
                <not_has_text text=" ivsm2ap2=" />
                <not_has_text text=" oms=" />
                <not_has_text text=" omsms=" />
                <not_has_text text=" oclean=" />
                <not_has_text text=" analyze " />
                <not_has_text text=" loss " />
                <not_has_text text=" intens " />
                <not_has_text text=" dbe " />
                <not_has_text text=" cm " />
                <not_has_text text=" pc " />
                <not_has_text text=" sc " />
            </assert_command>
        </test>
    </tests>
    <help><![CDATA[
        
**Overview**

Genform generates candidate molecular formulas from high-resolution
MS data. It calculates match values (MV) that show how well candidate molecular formulas fit the MS
isotope peak distributions (MS MV) and the high-resolution MS/MS fragment peak masses (MSMS
MV). Finally it computes a combined match value from these two scores. This software can be
regarded as a further development of the ElCoCo and MolForm modules of MOLGEN-MS with a clear
specialization towards MS/MS.

**Input**        

MS and MS/MS centroided spectra are given as tab separated files with two columns:

1. m/z
2. intensity

**Output**

Main output containing a list of all possible molecular formulas, further options can be chosen:

1. if the explained peaks shall also be written to the output
2. if the explanation shall be given as molecular formulas for the fragment or as neutral losses
3. double bond equivalents for each molecular formula
4. calculated ion masses 
5. output match values in percent

Output structure:

molecular fomula / (double bond equivalent) / (calculated mass) / ppm / MS MV / MSMS MV / combined MV 

ppm: The value ppm denotes the relative deviation of
the calculated mass of the molecular ion from the mass of the basepeak (basemass) in the MS in
parts per million (ppm).

MS MV: It is calculated from measured and
theoretical isotope peak intensities. The MV ranges from 0 to 1. The better theoretical and measured
isotope distributions fit, the higher is the MV. There are three different algorithms implemented for
calculating the MS MV, which are nomalized dot product, nomalized sum of squared errors and the normalized sum of absolute errors.

MSMS MV: It shows how well the masses of the MS/MS fit possible fragments of the candidate molecular formula. Again, the MS/MS MV is a value
between 0 and 1, whereby higher values reflect a better fit. 

combined MV: combined match value

Example output:

======== === =========== ====== ======== ========= ========
C4H9N3O2  2   132.07675   -0.8   96.674   100.000   96.674
======== === =========== ====== ======== ========= ========

**Advanced options**

For undersanding of the parameters and rules for creating possible molecular formulas, see `the manual of the commandline tool <https://depot.galaxyproject.org/software/molgen_manual/molgen_manual_1.0.1.5_src_all.pdf>`_ for info.

    ]]></help>

    <citations>
        <citation type="doi">10.3390/metabo3020440</citation>
        <citation type="bibtex">@article{meringer2011ms,
        title={MS/MS data improves automated determination of molecular formulas by mass spectrometry},
        author={Meringer, Markus and Reinker, Stefan and Zhang, Juan and Muller, Alban},
        journal={MATCH Commun. Math. Comput. Chem},
        volume={65},
        number={2},
        pages={259--290},
        year={2011}
        }</citation>
 </citations>
</tool>