## ----include=FALSE------------------------------------------------------------ # default chunk options knitr::opts_chunk$set(collapse = TRUE, message = FALSE, comment = "#>") ## ----------------------------------------------------------------------------- # load isoorbi library library(isoorbi) ## ----------------------------------------------------------------------------- # path to a test file included in the isoorbi package # this file contains data from multiple analyses for the same compound file_path <- system.file(package = "isoorbi", "extdata", "testfile_flow.isox") # load data df <- file_path |> # reads .isox test data orbi_read_isox() |> # optionally: keeps only most important columns; equivalent to simplify check box in IsoX orbi_simplify_isox() # If you wish to read a whole folder of files, you can use the orbi_find_isox() function # df <- "path_to_your_folder" |> orbi_find_isox() |> orbi_read_isox() ## ----------------------------------------------------------------------------- # check for noise and outliers df_flagged <- df |> orbi_flag_satellite_peaks() |> # removes minor signals that were reported by IsoX in the same tolerance window where the peak of interest is orbi_flag_weak_isotopocules(min_percent = 2) |> # removes signals of isotopocules that were not detected at least in min_percent scans orbi_flag_outliers(agc_fold_cutoff = 2) # removes outlying scans that have more than 2 times or less than 1/2 times the average number of ions in the Orbitrap analyzer; another method: agc_window (see function documentation for more details) ## ----------------------------------------------------------------------------- # define base peak and calculate the results table df_results <- df_flagged |> orbi_define_basepeak(basepeak_def = "M0")|> # sets one isotopocule in the dataset as the base peak (denominator) for ratio calculation orbi_summarize_results(ratio_method = "sum") # calculates ratios and generates a results table ## ----fig.width=6, fig.height=4------------------------------------------------ # use a ggplot to plot the data library(ggplot2) df_results |> ggplot() + aes( x = sprintf("%s/%s", isotopocule, basepeak), y = ratio, ymin = ratio - ratio_sem, ymax = ratio + ratio_sem, color = filename ) + geom_pointrange() + scale_color_brewer(palette = "Set1") + orbi_default_theme() + labs(x = NULL, y = "ratio") + facet_wrap(~compound)