%\VignetteIndexEntry{RI correction extra}
%\VignetteKeywords{preprocess, GC-MS, analysis}
%\VignettePackage{TargetSearch}
%\VignetteEngine{knitr::knitr}

\documentclass[a4paper]{article}

<<style-knitr, eval=TRUE, echo=FALSE, results="asis">>=
BiocStyle::latex()
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\usepackage{booktabs}
\definecolor{NoteBlue}{RGB}{45, 28, 156}
\newcommand{\Note}[2][Note]{\textsl{\textcolor{NoteBlue}{#1: #2}}}

\title{RI Correction Extra}
\author{\'Alvaro Cuadros-Inostroza}
\affil{Max Planck Institute for Molecular Plant Physiology, Potsdam, Germnay}

\begin{document}

\maketitle

\begin{abstract}
This vignette explains how to transform retention time (RT) to retention
index (RI) when the RI markers are not spiked in the samples. The
same principle can be applied to other uncommon scenarios.
\end{abstract}

\packageVersion{\Sexpr{BiocStyle::pkg_ver("TargetSearch")}}

Report issues on \url{https://github.com/acinostroza/TargetSearch/issues}

\newpage

\tableofcontents

\section{Motivation}

The \Biocpkg{TargetSearch} package assumes that the retention index
markers (RIM), such as n-alkanes or FAMEs (fatty acid methyl esters),
are injected together with the biological samples. A different approach
often used is to inject the RIMs separately, alternating between
RIMs and biological samples. For example, a GC-MS run may look like
the list shown in Table~\ref{tab:01}.

\begin{table}[ht]
\begin{tabular}{cc}
\toprule
Measurement Order & Sample Type \\
\midrule
1  & Alkanes \\
2  & Biological \\
3  & Biological \\
4  & Biological \\
5  & Biological \\
6  & Alkanes \\
7  & Biological \\
8  & Biological \\
9  & Biological \\
10 & Biological \\
11 & Alkanes \\
12 & Biological \\
13 & Biological \\
14 & Biological \\
15 & Biological \\
\bottomrule
\end{tabular}
\caption{An example of a GC-MS run. \emph{Alkanes} samples are retention
index markers (RIMs).}
\label{tab:01}
\end{table}

In the example (Table~\ref{tab:01}), samples 1, 6, 11 are RIMs and the rest
are biological samples. The assumption is that the retention time shifts
between consecutive runs are not significant, so sample \#1 is used to correct
samples \#2-5, sample \#6 corrects samples \#7-10, and so on. This document
shows how to perform RI correction in such case with \Biocpkg{TargetSearch}, using
the available chromatograms of the \Biocpkg{TargetSearchData} package.

\section{Retention Index correction}

First, we load the required packages
<<LibraryPreload>>=
library(TargetSearch)
library(TargetSearchData)
@

Specify the directory where the CDF files are. In this example we will
use the CDF files of \Biocpkg{TargetSearchData} package, which
can be retrieved by the function \Rfunction{tsd\_data\_path()}.

<<cdffiles>>=
cdfPath <- tsd_data_path()
dir(cdfPath, pattern="cdf$")
@

Create a \Robject{tsSample} object using all chromatograms from the
previous directory. Also set the RI path to the current directory.
Use the R commands \Rfunction{setwd()} and \Rfunction{getwd()}
to set/get the working directory.

<<samples>>=
samples.all <- ImportSamplesFromDir(cdfPath)
RIpath(samples.all) <- "."
@

Import retention index marker times limits. We will use the
ones defined in \Biocpkg{TargetSearchData} (the function
\Rfunction{tsd\_file\_path()} gets the full path of the file).

<<rim>>=
rimLimits <- ImportFameSettings(tsd_file_path("rimLimits.txt"))
rimLimits
@

Run the retention index correction methods. Here we use
an intensity threshold of 50,
the peak detection method is "ppc", and the time window width
is $2*15+1=31$ scan points (equal to 1.5 seconds in this example).

\Note{We skip conversion to CDF-4 files for illustration purposes.
In a normal workflow you would set \Rcode{writeCDF4path=TRUE} (the
default)}.

<<ricorrect>>=
RImatrix <- RIcorrect(samples.all, rimLimits, writeCDF4path=FALSE,
            Window=15, pp.method="ppc", IntThreshold=50)
RImatrix
@

Now you should make sure that the retention times (RT) of the RIMs
(not the biological samples!!) are correct by using a chromatogram
visualization tool such as LECO Pegasus, AMDIS, etc. Because there
are no RIMs inyected in the biological samples, the RTs of the
RIMs found in \Robject{RImatrix} will make no sense, so you don't need
to check them.

Up to now we have run the usual \Biocpkg{TargetSearch} workflow
which you can find in the main vignette. To correct the RI of
the biological samples, the following procedure can be used.

First, we need a logical vector indicating which samples are
RIMs and which biological. For example, you could create a
vector like this.

<<isRIMarker>>=
isRIMarker <- rep(FALSE, length(samples.all))
isRIMarker[c(1, 6, 11)] <- TRUE
@

where \Robject{isRIMarker} is a logical vector of length equal
to the number of samples in which \Rcode{TRUE} signals that the
respective sample is a RIM and \Rcode{FALSE} otherwise. Note that here
we have set components 1, 6, 11 to \Rcode{TRUE} just like
the example in Table~\ref{tab:01}.

Then we have to copy the RIM columns of \Robject{RImatrix} to
their respective biological sample columns. In other words,
copy column 1 to columns 2, 3, 4, 5; column 6 to columns 7,
8, 9, 10; and so on. There are many ways to achieve that,
here I show two examples.

<<updateRImatrix2>>=
RImatrix2 <- RImatrix
RImatrix2[, 2:5]   <- RImatrix[,1]
RImatrix2[, 7:10]  <- RImatrix[,6]
RImatrix2[, 12:15] <- RImatrix[,11]
@

This code snippet is a straight forward method, but has the
disadvantage that the column indexes must be filled manually.
A more general approach could be.

<<updateRImatrix>>=
RImatrix2 <- RImatrix
z <- cumsum(as.numeric(isRIMarker))
for(i in unique(z))
    RImatrix2[, z==i] <- RImatrix[, z==i][,1]
RImatrix2
@

After the \Robject{RImatrix2} object is corrected, the RI files
of the biological samples must be fixed as well.

<<fixRI>>=
fixRI(samples.all, rimLimits, RImatrix2, which(!isRIMarker))
@

Finally, we remove the standards, since we don't need them anymore.

<<finally>>=
samples <- samples.all[!isRIMarker]
RImatrix <- RImatrix2[, !isRIMarker]
@

After that, we can continue we the normal \Biocpkg{TargetSearch}
workflow.

A script with all the commands used in this document is located
in the \file{doc} directory of the \Biocpkg{TargetSearch} package.

\section{Session info}

Output of \Rfunction{sessionInfo()} on the system on which
this document was compiled.

<<sessionInfo, results="asis", echo=FALSE>>=
toLatex(sessionInfo())
@

\end{document}
