Data container storing xcms preprocessing results

The XCMSnExp object is a container for the results of a G/LC-MS data preprocessing that comprises chromatographic peak detection, alignment and correspondence. These results can be accessed with the chromPeaks, adjustedRtime and featureDefinitions functions; see below (after the Usage, Arguments, Value and Slots sections) for more details). Along with the results, the object contains the processing history that allows to track each processing step along with the used settings. This can be extracted with the processHistory method. XCMSnExp objects, by directly extending the OnDiskMSnExp object from the MSnbase package, inherit all of its functionality and allows thus an easy access to the full raw data at any stage of an analysis. To support interaction with packages requiring the old objects, XCMSnExp objects can be coerced into xcmsSet objects using the as method (see examples below). All preprocessing results will be passed along to the resulting xcmsSet object.

General functions for XCMSnExp objects are (see further below for specific function to handle chromatographic peak data, alignment and correspondence results):

processHistoryTypes returns the available types of process histories. These can be passed with argument type to the processHistory method to extract specific process step(s).

hasFilledChromPeaks: whether filled-in peaks are present or not.

profMat: creates a profile matrix, which is a n x m matrix, n (rows) representing equally spaced m/z values (bins) and m (columns) the retention time of the corresponding scans. Each cell contains the maximum intensity measured for the specific scan and m/z values. See profMat for more details and description of the various binning methods.

hasAdjustedRtime: whether the object provides adjusted retention times.

hasFeatures: whether the object contains correspondence results (i.e. features).

hasChromPeaks: whether the object contains peak detection results.

hasFilledChromPeaks: whether the object contains any filled-in chromatographic peaks.

adjustedRtime,adjustedRtime<-: extract/set adjusted retention times. adjustedRtime<- should not be called manually, it is called internally by the adjustRtime methods. For XCMSnExp objects, adjustedRtime<- does also apply retention time adjustments to eventually present chromatographic peaks. The bySample parameter allows to specify whether the adjusted retention time should be grouped by sample (file).

featureDefinitions, featureDefinitions<-: extract or set the correspondence results, i.e. the mz-rt features (peak groups). Similar to the chromPeaks it is possible to extract features for specified m/z and/or rt ranges. The function supports also the parameter type that allows to specify which features to be returned if any of rt or mz is specified. For details see help of chromPeaks. See also featureSummary for a function to calculate simple feature summaries.

chromPeaks, chromPeaks<-: extract or set the matrix containing the information on identified chromatographic peaks. Rownames of the matrix represent unique IDs of the respective peaks within the experiment. Parameter bySample allows to specify whether peaks should be returned ungrouped (default bySample = FALSE) or grouped by sample (bySample = TRUE). The chromPeaks<- method for XCMSnExp objects removes also all correspondence (peak grouping) and retention time correction (alignment) results. The optional arguments rt, mz, ppm and type allow to extract only chromatographic peaks overlapping the defined retention time and/or m/z ranges. Argument type allows to define how overlapping is determined: for type == "any" (the default), all peaks that are even partially overlapping the region are returned (i.e. for which either "mzmin" or "mzmax" of the chromPeaks or featureDefinitions matrix are within the provided m/z range), for type == "within" the full peak has to be within the region (i.e. both "mzmin" and "mzmax" have to be within the m/z range) and for type == "apex_within" the peak's apex position (highest signal of the peak) has to be within the region (i.e. the peak's or features m/z has to be within the m/z range). See description of the return value for details on the returned matrix. Users usually don't have to use the chromPeaks<- method directly as detected chromatographic peaks are added to the object by the findChromPeaks method. Also, chromPeaks<- will replace any existing chromPeakData.

chromPeakData and chromPeakData<- allow to get or set arbitrary chromatographic peak annotations. These are returned or ar returned as a DataFrame. Note that the number of rows and the rownames of the DataFrame have to match those of chromPeaks.

rtime: extracts the retention time for each scan. The bySample parameter allows to return the values grouped by sample/file and adjusted whether adjusted or raw retention times should be returned. By default the method returns adjusted retention times, if they are available (i.e. if retention times were adjusted using the adjustRtime method).

mz: extracts the mz values from each scan of all files within an XCMSnExp object. These values are extracted from the original data files and eventual processing steps are applied on the fly. Using the bySample parameter it is possible to switch from the default grouping of mz values by spectrum/scan to a grouping by sample/file.

intensity: extracts the intensity values from each scan of all files within an XCMSnExp object. These values are extracted from the original data files and eventual processing steps are applied on the fly. Using the bySample parameter it is possible to switch from the default grouping of intensity values by spectrum/scan to a grouping by sample/file.

spectra: extracts the Spectrum objects containing all data from object. The values are extracted from the original data files and eventual processing steps are applied on the fly. By setting bySample = TRUE, the spectra are returned grouped by sample/file. If the XCMSnExp object contains adjusted retention times, these are returned by default in the Spectrum objects (can be overwritten by setting adjusted = FALSE).

processHistory: returns a list of ProcessHistory objects (or objects inheriting from this base class) representing the individual processing steps that have been performed, eventually along with their settings (Param parameter class). Optional arguments fileIndex, type and msLevel allow to restrict to process steps of a certain type or performed on a certain file or MS level.

dropChromPeaks: drops any identified chromatographic peaks and returns the object without that information. Note that for XCMSnExp objects the method drops by default also results from a correspondence (peak grouping) analysis. Adjusted retention times are removed if the alignment has been performed after peak detection. This can be overruled with keepAdjustedRtime = TRUE.

dropFeatureDefinitions: drops the results from a correspondence (peak grouping) analysis, i.e. the definition of the mz-rt features and returns the object without that information. Note that for XCMSnExp objects the method will also by default drop retention time adjustment results, if these were performed after the last peak grouping (i.e. which base on the results from the peak grouping that are going to be removed). All related process history steps are removed too as well as eventually filled in peaks (by fillChromPeaks). The parameter keepAdjustedRtime can be used to avoid removal of adjusted retention times.

dropAdjustedRtime: drops any retention time adjustment information and returns the object without adjusted retention time. For XCMSnExp objects, this also reverts the retention times reported for the chromatographic peaks in the peak matrix to the original, raw, ones (after chromatographic peak detection). Note that for XCMSnExp objects the method drops also all peak grouping results if these were performed after the retention time adjustment. All related process history steps are removed too.

findChromPeaks performs chromatographic peak detection on the provided XCMSnExp objects. For more details see the method for XCMSnExp. Note that by default (with parameter add = FALSE) previous peak detection results are removed. Use add = TRUE to perform a second round of peak detection and add the newly identified peaks to the previous peak detection results. Correspondence results (features) are always removed prior to peak detection. Previous alignment (retention time adjustment) results are kept, i.e. chromatographic peak detection is performed using adjusted retention times if the data was first aligned using e.g. obiwarp (adjustRtime).

dropFilledChromPeaks: drops any filled-in chromatographic peaks (filled in by the fillChromPeaks method) and all related process history steps.

spectrapply applies the provided function to each Spectrum in the object and returns its results. If no function is specified the function simply returns the list of Spectrum objects.

XCMSnExp objects can be combined with the c function. This combines identified chromatographic peaks and the objects' pheno data but discards alignment results or feature definitions.

plot plots the spectrum data (see plot for MSnExp objects in the MSnbase package for more details. For type = "XIC", identified chromatographic peaks will be indicated as rectangles with border color peakCol.

Usage

processHistoryTypes()

# S4 method for class 'XCMSnExp'
hasFilledChromPeaks(object)

# S4 method for class 'OnDiskMSnExp'
profMat(
  object,
  method = "bin",
  step = 0.1,
  baselevel = NULL,
  basespace = NULL,
  mzrange. = NULL,
  fileIndex,
  ...
)

# S4 method for class 'XCMSnExp'
show(object)

# S4 method for class 'XCMSnExp'
hasAdjustedRtime(object)

# S4 method for class 'XCMSnExp'
hasFeatures(object, msLevel = integer())

# S4 method for class 'XCMSnExp'
hasChromPeaks(object, msLevel = integer())

# S4 method for class 'XCMSnExp'
hasFilledChromPeaks(object)

# S4 method for class 'XCMSnExp'
adjustedRtime(object, bySample = FALSE)

# S4 method for class 'XCMSnExp'
adjustedRtime(object) <- value

# S4 method for class 'XCMSnExp'
featureDefinitions(
  object,
  mz = numeric(),
  rt = numeric(),
  ppm = 0,
  type = c("any", "within", "apex_within"),
  msLevel = integer()
)

# S4 method for class 'XCMSnExp'
featureDefinitions(object) <- value

# S4 method for class 'XCMSnExp'
chromPeaks(
  object,
  bySample = FALSE,
  rt = numeric(),
  mz = numeric(),
  ppm = 0,
  msLevel = integer(),
  type = c("any", "within", "apex_within"),
  isFilledColumn = FALSE
)

# S4 method for class 'XCMSnExp'
chromPeaks(object) <- value

# S4 method for class 'XCMSnExp'
rtime(object, bySample = FALSE, adjusted = hasAdjustedRtime(object))

# S4 method for class 'XCMSnExp'
mz(object, bySample = FALSE, BPPARAM = bpparam())

# S4 method for class 'XCMSnExp'
intensity(object, bySample = FALSE, BPPARAM = bpparam())

# S4 method for class 'XCMSnExp'
spectra(
  object,
  bySample = FALSE,
  adjusted = hasAdjustedRtime(object),
  BPPARAM = bpparam()
)

# S4 method for class 'XCMSnExp'
processHistory(object, fileIndex, type, msLevel)

# S4 method for class 'XCMSnExp'
dropChromPeaks(object, keepAdjustedRtime = FALSE)

# S4 method for class 'XCMSnExp'
dropFeatureDefinitions(object, keepAdjustedRtime = FALSE, dropLastN = -1)

# S4 method for class 'XCMSnExp'
dropAdjustedRtime(object)

# S4 method for class 'XCMSnExp'
profMat(
  object,
  method = "bin",
  step = 0.1,
  baselevel = NULL,
  basespace = NULL,
  mzrange. = NULL,
  fileIndex,
  ...
)

# S4 method for class 'XCMSnExp,Param'
findChromPeaks(
  object,
  param,
  BPPARAM = bpparam(),
  return.type = "XCMSnExp",
  msLevel = 1L,
  add = FALSE
)

# S4 method for class 'XCMSnExp'
dropFilledChromPeaks(object)

# S4 method for class 'XCMSnExp'
spectrapply(object, FUN = NULL, BPPARAM = bpparam(), ...)

# S3 method for class 'XCMSnExp'
c(...)

# S4 method for class 'XCMSnExp'
chromPeakData(object, ...)

# S4 method for class 'XCMSnExp'
chromPeakData(object) <- value

# S4 method for class 'XCMSnExp,missing'
plot(x, y, type = c("spectra", "XIC"), peakCol = "#ff000060", ...)

Arguments

object

For adjustedRtime, featureDefinitions, chromPeaks, hasAdjustedRtime, hasFeatures and hasChromPeaks either a MsFeatureData or a XCMSnExp object, for all other methods a XCMSnExp object.

method

character(1) defining the profile matrix generation method. Allowed are "bin", "binlin", "binlinbase" and "intlin". See details section for more information.

step

numeric(1) representing the m/z bin size.

baselevel

numeric(1) representing the base value to which empty elements (i.e. m/z bins without a measured intensity) should be set. Only considered if method = "binlinbase". See baseValue parameter of imputeLinInterpol() for more details.

basespace

numeric(1) representing the m/z length after which the signal will drop to the base level. Linear interpolation will be used between consecutive data points falling within 2 * basespace to each other. Only considered if method = "binlinbase". If not specified, it defaults to 0.075. Internally this parameter is translated into the distance parameter of the imputeLinInterpol() function by distance = floor(basespace / step). See distance parameter of imputeLinInterpol() for more details.

mzrange.

Optional numeric(2) manually specifying the mz value range to be used for binnind. If not provided, the whole m/z value range is used.

fileIndex

For processHistory: optional integer specifying the index of the files/samples for which the ProcessHistory objects should be retrieved.

...

Additional parameters.

msLevel

integer specifying the MS level(s) for which identified chromatographic peaks should be returned.

bySample

logical(1) specifying whether results should be grouped by sample.

value

For adjustedRtime<-: a list (length equal to the number of samples) with numeric vectors representing the adjusted retention times per scan.

For featureDefinitions<-: a DataFrame with peak grouping information. See return value for the featureDefinitions method for the expected format.

For chromPeaks<-: a matrix with information on detected peaks. See return value for the chromPeaks method for the expected format.

mz

optional numeric(2) defining the mz range for which chromatographic peaks should be returned.

rt

optional numeric(2) defining the retention time range for which chromatographic peaks should be returned.

ppm

optional numeric(1) specifying the ppm by which the mz range should be extended. For a value of ppm = 10, all peaks within mz[1] - ppm / 1e6 and mz[2] + ppm / 1e6 are returned.

type

For processHistory: restrict returned ProcessHistory objects to analysis steps of a certain type. Use the processHistoryTypes to list all supported values. For chromPeaks: character specifying which peaks to return if rt or mz are defined. For type = "any" all chromatographic peaks partially overlapping the range defined by mz and/or rt are returned, type = "within" returns only peaks completely within the region and type = "apex_within" peaks for which the peak's apex is within the region.

isFilledColumn

logical(1) whether a column "is_filled" is included in the returned "matrix" providing the information if a peak was filled in. Alternatively, this information would be provided by the chromPeakData data frame.

adjusted

logical(1) whether adjusted or raw (i.e. the original retention times reported in the files) should be returned.

BPPARAM

Parameter class for parallel processing. See bpparam.

keepAdjustedRtime

For dropFeatureDefinitions,XCMSnExp: logical(1) defining whether eventually present retention time adjustment should not be dropped. By default dropping feature definitions drops retention time adjustment results too.

dropLastN

For dropFeatureDefinitions,XCMSnExp: numeric(1) defining the number of peak grouping related process history steps to remove. By default dropLastN = -1, dropping the chromatographic peaks removes all process history steps related to peak grouping. Setting e.g. dropLastN = 1 will only remove the most recent peak grouping related process history step.

param

A CentWaveParam, MatchedFilterParam, MassifquantParam, MSWParam or CentWavePredIsoParam object with the settings for the chromatographic peak detection algorithm.

return.type

Character specifying what type of object the method should return. Can be either "XCMSnExp" (default), "list" or "xcmsSet".

add

For findChromPeaks: if newly identified chromatographic peaks should be added to the peak matrix with the already identified chromatographic peaks. By default (add = FALSE) previous peak detection results will be removed.

FUN

For spectrapply: a function that should be applied to each spectrum in the object.

x

For plot: XCMSnExp object.

y

For plot: not used.

peakCol

For plot: the color that should be used to indicate identified chromatographic peaks (only in combination with type = "XIC" and if chromatographic peaks are present).

Value

For profMat: a list with a the profile matrix matrix (or matrices if fileIndex was not specified or if length(fileIndex) > 1). See profile-matrix for general help and information about the profile matrix.

For adjustedRtime: if bySample = FALSE a numeric vector with the adjusted retention for each spectrum of all files/samples within the object. If bySample = TRUE a list (length equal to the number of samples) with adjusted retention times grouped by sample. Returns NULL if no adjusted retention times are present.

For featureDefinitions: a DataFrame with peak grouping information, each row corresponding to one mz-rt feature (grouped peaks within and across samples) and columns "mzmed" (median mz value), "mzmin" (minimal mz value), "mzmax" (maximum mz value), "rtmed" (median retention time), "rtmin" (minimal retention time), "rtmax" (maximal retention time) and "peakidx". Column "peakidx" contains a list with indices of chromatographic peaks (rows) in the matrix returned by the chromPeaks method that belong to that feature group. The method returns NULL if no feature definitions are present. featureDefinitions supports also parameters mz, rt, ppm and type to return only features within certain ranges (see description of chromPeaks for details).

For chromPeaks: if bySample = FALSE a matrix (each row being a chromatographic peak, rownames representing unique IDs of the peaks) with at least the following columns: "mz" (intensity-weighted mean of mz values of the peak across scans/retention times), "mzmin" (minimal mz value), "mzmax" (maximal mz value), "rt" (retention time of the peak apex), "rtmin" (minimal retention time), "rtmax" (maximal retention time), "into" (integrated, original, intensity of the peak), "maxo" (maximum intentity of the peak), "sample" (sample index in which the peak was identified) and Depending on the employed peak detection algorithm and the verboseColumns parameter of it, additional columns might be returned. If parameter isFilledColumn was set to TRUE a column named "is_filled" is also returned. For bySample = TRUE the chromatographic peaks are returned as a list of matrices, each containing the chromatographic peaks of a specific sample. For samples in which no peaks were detected a matrix with 0 rows is returned.

For rtime: if bySample = FALSE a numeric vector with the retention times of each scan, if bySample = TRUE a list of numeric vectors with the retention times per sample.

For mz: if bySample = FALSE a list with the mz values (numeric vectors) of each scan. If bySample = TRUE a list with the mz values per sample.

For intensity: if bySample = FALSE a list with the intensity values (numeric vectors) of each scan. If bySample = TRUE a list with the intensity values per sample.

For spectra: if bySample = FALSE a list with Spectrum objects. If bySample = TRUE the result is grouped by sample, i.e. as a list of lists, each element in the outer list being the list of spectra of the specific file.

For processHistory: a list of ProcessHistory objects providing the details of the individual data processing steps that have been performed.

Slots

.processHistory: list with XProcessHistory objects tracking all individual analysis steps that have been performed.
msFeatureData: MsFeatureData class extending environment and containing the results from a chromatographic peak detection (element "chromPeaks"), peak grouping (element "featureDefinitions") and retention time correction (element "adjustedRtime") steps. This object should not be manipulated directly.

Note

The "chromPeaks" element in the msFeatureData slot is equivalent to the @peaks slot of the xcmsSet object, the "featureDefinitions" contains information from the @groups and @groupidx slots from an xcmsSet object.

Chromatographic peak data

Chromatographic peak data is added to an XCMSnExp object by the findChromPeaks function. Functions to access chromatographic peak data are:

hasChromPeaks whether chromatographic peak data is available, see below for help of the function.
chromPeaks access chromatographic peaks (see below for help).
dropChromPeaks remove chromatographic peaks (see below for help).
dropFilledChromPeaks remove filled-in peaks (see below for help).
fillChromPeaks fill-in missing peaks (see respective help page).
plotChromPeaks plot identified peaks for a file (see respective help page).
plotChromPeakImage plot distribution of peaks along the retention time axis (see respective help page).
highlightChromPeaks add chromatographic peaks to an existing plot of a Chromatogram (see respective help page).

Adjusted retention times

Adjusted retention times are stored in an XCMSnExp object besides the original, raw, retention times, allowing to switch between raw and adjusted times. It is also possible to replace the raw retention times with the adjusted ones with the applyAdjustedRtime. The adjusted retention times are added to an XCMSnExp by the adjustRtime function. All functions related to the access of adjusted retention times are:

hasAdjustedRtime whether adjusted retention times are available (see below for help).
dropAdjustedRtime remove adjusted retention times (see below for help).
applyAdjustedRtime replace the raw retention times with the adjusted ones (see respective help page).
plotAdjustedRtime plot differences between adjusted and raw retention times (see respective help page).

Correspondence results, features

The correspondence analysis (groupChromPeaks) adds the feature definitions to an XCMSnExp object. All functions related to these are listed below:

hasFeatures whether correspondence results are available (see below for help).
featureDefinitions access the definitions of the features (see below for help).
dropFeatureDefinitions remove correspondence results (see below for help).
featureValues access values for features (see respective help page).
featureSummary perform a simple summary of the defined features (see respective help page).
overlappingFeatures identify features that are overlapping or close in the m/z - rt space (see respective help page).
quantify extract feature intensities and put them, along with feature definitions and phenodata information, into a SummarizedExperiment. See help page for details.

Author

Johannes Rainer

Examples


## Load a test data set with detected peaks
library(MSnbase)
#> Loading required package: BiocGenerics
#> Loading required package: generics
#> 
#> Attaching package: ‘generics’
#> The following objects are masked from ‘package:base’:
#> 
#>     as.difftime, as.factor, as.ordered, intersect, is.element, setdiff,
#>     setequal, union
#> 
#> Attaching package: ‘BiocGenerics’
#> The following objects are masked from ‘package:stats’:
#> 
#>     IQR, mad, sd, var, xtabs
#> The following objects are masked from ‘package:base’:
#> 
#>     Filter, Find, Map, Position, Reduce, anyDuplicated, aperm, append,
#>     as.data.frame, basename, cbind, colnames, dirname, do.call,
#>     duplicated, eval, evalq, get, grep, grepl, is.unsorted, lapply,
#>     mapply, match, mget, order, paste, pmax, pmax.int, pmin, pmin.int,
#>     rank, rbind, rownames, sapply, saveRDS, table, tapply, unique,
#>     unsplit, which.max, which.min
#> Loading required package: Biobase
#> Welcome to Bioconductor
#> 
#>     Vignettes contain introductory material; view with
#>     'browseVignettes()'. To cite Bioconductor, see
#>     'citation("Biobase")', and for packages 'citation("pkgname")'.
#> Loading required package: mzR
#> Loading required package: Rcpp
#> Loading required package: S4Vectors
#> Loading required package: stats4
#> 
#> Attaching package: ‘S4Vectors’
#> The following object is masked from ‘package:utils’:
#> 
#>     findMatches
#> The following objects are masked from ‘package:base’:
#> 
#>     I, expand.grid, unname
#> 
#> This is MSnbase version 2.33.2 
#>   Visit https://lgatto.github.io/MSnbase/ to get started.
#>  Consider switching to the 'R for Mass Spectrometry'
#>  packages - see https://RforMassSpectrometry.org for details.
#> 
#> Attaching package: ‘MSnbase’
#> The following object is masked from ‘package:base’:
#> 
#>     trimws
data(faahko_sub)
## Update the path to the files for the local system
dirname(faahko_sub) <- system.file("cdf/KO", package = "faahKO")

## Disable parallel processing for this example
register(SerialParam())

## The results from the peak detection are now stored in the XCMSnExp
## object
faahko_sub
#> MSn experiment data ("XCMSnExp")
#> Object size in memory: 1.22 Mb
#> - - - Spectra data - - -
#>  MS level(s): 1 
#>  Number of spectra: 3834 
#>  MSn retention times: 41:41 - 74:60 minutes
#> - - - Processing information - - -
#> Data loaded [Thu Mar 14 11:42:34 2024] 
#>  MSnbase version: 2.29.3 
#> - - - Meta data  - - -
#> phenoData
#>   rowNames: ko15.CDF ko16.CDF ko18.CDF
#>   varLabels: sampleNames
#>   varMetadata: labelDescription
#> Loaded from:
#>   [1] ko15.CDF...  [3] ko18.CDF
#>   Use 'fileNames(.)' to see all files.
#> protocolData: none
#> featureData
#>   featureNames: F1.S0001 F1.S0002 ... F3.S1278 (3834 total)
#>   fvarLabels: fileIdx spIdx ... spectrum (35 total)
#>   fvarMetadata: labelDescription
#> experimentData: use 'experimentData(object)'
#> - - - xcms preprocessing - - -
#> Chromatographic peak detection:
#>  method: centWave 
#>  248 peaks identified in 3 samples.
#>  On average 82.7 chromatographic peaks per sample.

## The detected peaks can be accessed with the chromPeaks method.
head(chromPeaks(faahko_sub))
#>          mz mzmin mzmax       rt    rtmin    rtmax       into       intb   maxo
#> CP001 453.2 453.2 453.2 2506.073 2501.378 2527.982  1007409.0  1007380.8  38152
#> CP002 302.0 302.0 302.0 2617.185 2595.275 2640.659   687146.6   671297.8  30552
#> CP003 344.0 344.0 344.0 2679.783 2646.919 2709.517  5210015.9  5135916.9 152320
#> CP004 430.1 430.1 430.1 2681.348 2639.094 2712.647  2395840.3  2299899.6  65752
#> CP005 366.0 366.0 366.0 2679.783 2642.224 2718.907  3365174.0  3279468.3  79928
#> CP006 343.0 343.0 343.0 2678.218 2637.529 2712.647 24147443.2 23703761.7 672064
#>          sn sample
#> CP001 38151      1
#> CP002    46      1
#> CP003    68      1
#> CP004    42      1
#> CP005    49      1
#> CP006    87      1

## The settings of the chromatographic peak detection can be accessed with
## the processHistory method
processHistory(faahko_sub)
#> [[1]]
#> Object of class "XProcessHistory"
#>  type: Peak detection 
#>  date: Thu Mar 14 11:42:34 2024 
#>  info:  
#>  fileIndex: 1,2,3 
#>  Parameter class: CentWaveParam 
#>  MS level(s) 1 
#> 

## Also the parameter class for the peak detection can be accessed
processParam(processHistory(faahko_sub)[[1]])
#> Object of class:  CentWaveParam 
#>  Parameters:
#>  - ppm: [1] 25
#>  - peakwidth: [1] 20 50
#>  - snthresh: [1] 40
#>  - prefilter: [1]     3 10000
#>  - mzCenterFun: [1] "wMean"
#>  - integrate: [1] 1
#>  - mzdiff: [1] -0.001
#>  - fitgauss: [1] FALSE
#>  - noise: [1] 10000
#>  - verboseColumns: [1] FALSE
#>  - roiList: list()
#>  - firstBaselineCheck: [1] TRUE
#>  - roiScales: numeric(0)
#>  - extendLengthMSW: [1] FALSE
#>  - verboseBetaColumns: [1] FALSE

## The XCMSnExp inherits all methods from the pSet and OnDiskMSnExp classes
## defined in Bioconductor's MSnbase package. To access the (raw) retention
## time for each spectrum we can use the rtime method. Setting bySample = TRUE
## would cause the retention times to be grouped by sample
head(rtime(faahko_sub))
#> F1.S0001 F1.S0002 F1.S0003 F1.S0004 F1.S0005 F1.S0006 
#> 2501.378 2502.943 2504.508 2506.073 2507.638 2509.203 

## Similarly it is possible to extract the mz values or the intensity values
## using the mz and intensity method, respectively, also with the option to
## return the results grouped by sample instead of the default, which is
## grouped by spectrum. Finally, to extract all of the data we can use the
## spectra method which returns Spectrum objects containing all raw data.
## Note that all these methods read the information from the original input
## files and subsequently apply eventual data processing steps to them.
mzs <- mz(faahko_sub, bySample = TRUE)
length(mzs)
#> [1] 3
lengths(mzs)
#>      1      2      3 
#> 445769 440794 442516 

## The full data could also be read using the spectra data, which returns
## a list of Spectrum object containing the mz, intensity and rt values.
## spctr <- spectra(faahko_sub)
## To get all spectra of the first file we can split them by file
## head(split(spctr, fromFile(faahko_sub))[[1]])

############
## Filtering
##
## XCMSnExp objects can be filtered by file, retention time, mz values or
## MS level. For some of these filter preprocessing results (mostly
## retention time correction and peak grouping results) will be dropped.
## Below we filter the XCMSnExp object by file to extract the results for
## only the second file.
xod_2 <- filterFile(faahko_sub, file = 2)
xod_2
#> MSn experiment data ("XCMSnExp")
#> Object size in memory: 0.48 Mb
#> - - - Spectra data - - -
#>  MS level(s): 1 
#>  Number of spectra: 1278 
#>  MSn retention times: 41:41 - 74:60 minutes
#> - - - Processing information - - -
#> Data loaded [Thu Mar 14 11:42:34 2024] 
#> Filter: select file(s) 2. [Sun Dec 15 12:36:15 2024] 
#>  MSnbase version: 2.29.3 
#> - - - Meta data  - - -
#> phenoData
#>   rowNames: ko16.CDF
#>   varLabels: sampleNames
#>   varMetadata: labelDescription
#> Loaded from:
#>   ko16.CDF 
#> protocolData: none
#> featureData
#>   featureNames: F2.S0001 F2.S0002 ... F2.S1278 (1278 total)
#>   fvarLabels: fileIdx spIdx ... spectrum (35 total)
#>   fvarMetadata: labelDescription
#> experimentData: use 'experimentData(object)'
#> - - - xcms preprocessing - - -
#> Chromatographic peak detection:
#>  method: centWave 
#>  100 peaks identified in 1 samples.
#>  On average 100 chromatographic peaks per sample.

## Now the objects contains only the idenfified peaks for the second file
head(chromPeaks(xod_2))
#>          mz mzmin mzmax       rt    rtmin    rtmax       into        intb
#> CP088 384.1 384.1 384.1 2603.100 2592.145 2614.054   274906.3   274885.96
#> CP089 533.3 533.3 533.3 2686.042 2681.347 2689.172    70456.3    70450.04
#> CP090 344.0 344.0 344.0 2686.042 2648.483 2714.211  5700652.0  5574744.87
#> CP091 366.0 366.0 366.0 2686.042 2639.094 2725.166  3448376.1  3338483.07
#> CP092 343.0 343.0 343.0 2686.042 2645.353 2717.341 26229546.1 25615117.17
#> CP093 365.0 365.0 365.0 2686.042 2639.094 2726.731 15565868.0 14884380.84
#>         maxo    sn sample
#> CP088  15187 15186      1
#> CP089  11819 11818      1
#> CP090 151744    87      1
#> CP091  77640    42      1
#> CP092 692352    68      1
#> CP093 345536    48      1

##########
## Coercing to an xcmsSet object
##
## We can also coerce the XCMSnExp object into an xcmsSet object:
xs <- as(faahko_sub, "xcmsSet")
#> Note: you might want to set/adjust the 'sampclass' of the returned xcmSet object before proceeding with the analysis.
head(peaks(xs))
#>          mz mzmin mzmax       rt    rtmin    rtmax       into       intb   maxo
#> CP001 453.2 453.2 453.2 2506.073 2501.378 2527.982  1007409.0  1007380.8  38152
#> CP002 302.0 302.0 302.0 2617.185 2595.275 2640.659   687146.6   671297.8  30552
#> CP003 344.0 344.0 344.0 2679.783 2646.919 2709.517  5210015.9  5135916.9 152320
#> CP004 430.1 430.1 430.1 2681.348 2639.094 2712.647  2395840.3  2299899.6  65752
#> CP005 366.0 366.0 366.0 2679.783 2642.224 2718.907  3365174.0  3279468.3  79928
#> CP006 343.0 343.0 343.0 2678.218 2637.529 2712.647 24147443.2 23703761.7 672064
#>          sn sample
#> CP001 38151      1
#> CP002    46      1
#> CP003    68      1
#> CP004    42      1
#> CP005    49      1
#> CP006    87      1