2. Usage

This Chapter deals with some features of pymzml and explains the basic usage.

2.1. Basic usage

The Python scripting language is extended by pymzML to enable rapid evaluation, prototyping and even complex evaluation of large mass spectrometry datasets. The following examples are executed within the Python console (indicated by “>>>” ) but can equally be incorporated in standalone scripts.

The pymzML file object is declared as follows:

>>> import pymzml
>>> msrun = pymzml.run.Reader("big-1.0.0.mzML")

The input files can be plain or compressed mzML files. Initialization of the Run class accepts additional keywords, i.e. precision of MS1 or MSn runs and extra accessions (see: obo). The run object returns an iterator, hence one can loop over all spectra and/or chromatograms using classic Python syntax. Additionally, one can retrieve a specific spectrum by its nativeID via random access using msrun[1], msrun[‘TIC’] to access the chromatogram or in case of MRM experiments as e.g. msrun[‘transition_445-672’]. Note that random access requires the mzML file not to be compressed or truncated by a conversion program.

>>> for spectrum in msrun:
>>>   if spectrum['ms level'] == 2:

>>> spectrum_with_nativeID_100 = msrun[100]

Each iteration returns a spectrum object (spec.Spectrum) which offers basic information on the spectrum. The information can be accessed like a Python dictionary. The keys in this dictionary are the accession numbers (e.g. MS:1000511) or the name of the accession (e.g. “ms level”), as they are defined by the HUPO Proteomics Standards Initiative in the mzML vocabulary, i.e. in the OBO files. Both keys of the spectrum dictionary point to the value that has been extracted from the mzML file (see: obo). Association of MS tag to name is done on the basis of the OBO files which are supplied in the pymzML package. The pymzML package offers a simple script (see: queryOBO) that can be used to translate between MS tags and names. Examples for accessing MS tags by their names can be found here: OBO parser Class. It is worth noting that the definition of MS tags and their attribute that the user wants to be associated with the tag and the trivial name is a feature. For example, the scan time is associated with two values, the actual time and its unit. From a programming point of view, access to the time as float by calling spectrum[‘scan time’] is desirable. Defining those attributes and their value one is interested in, facilitates live ;)

Data that is associated with the current spectrum, i.e. mass over charge (m/z) or intensity values can be accessed by the .mz (spec.Spectrum.mz) or .intensity (spec.Spectrum.i) properties, respectively, which are iterators themselves. The .peaks (spec.Spectrum.peaks) property also offers an iterator which returns mz and intensity as a tuple for each peak:

>>>     for mz, i in spectrum.peaks:
>>>       print(mz, i)

Since mass spectrometry data can be measured in profile mode, the .centroidedPeak (spec.Spectrum.centroidedPeaks) property offers an iterator which performs a simple Gauss fit on the profiled data, thereby returning the fitted m/z and maximum intensity of the bell shape curve as tuple. Basic visualization of spectra using XHTML and SVG can be done using the plot submodule.

2.2. Advanced usage

The spectrum class also offers other functions, such as deconvolution, estimation of similarity of spectra, simple noise reduction, addition of spectra and simple arithmetics on the intensity values by multiplication or division. An averaged spectrum can be created during parsing as follows:

>>> t = pymzml.spec.Spectrum()
>>> for s in msrun:
>>>   if s['ms level'] == 1:
>>>     t += s / s['total ion current']

The addition of spectra is done by creating Gaussian distributions around the centroided peak. This reprofiling is done internally as part of the addition and its values can be accessed by the .reprofiledPeaks (spec.Spectrum.reprofiledPeaks) property of the spectrum. To reduce the computational overhead one can remove noise by calling the .removeNoise() (spec.Spectrum.removeNoise()) method of the spectrum class. This method accepts different modes of noise reduction. Deconvoluted peaks can be accessed for high precision spectra by calling the .deconvolutedPeaks (spec.Spectrum.deconvolutedPeaks) property. The pymzML package contains detailed example scripts for all these methods. ( Example scripts )