Abstract
A typical metabolomic analysis consists of a multi-step procedure. Variation can be introduced in any analysis segment if proper care in quality assurance is not taken, thus compromising the final results. Sample stability is one of those factors. Although sophisticated studies addressing sample decay over time have been performed in the medical field, they are emerging in plant metabolomics. Here, we focus on the stability of wheat floret extracts on queue inside an auto-injector held at 25 °C. The objective was to locate an analytical time window from extraction to injection with no significant difference occurring in the sample. Total ion current chromatograms, principal component analysis, and volcano plots were used to measure changes in the samples. Results indicate a maximum work window time of 7:45 h for Steele-ND wheat methanolic extractions in an auto-sampler at 25 °C. Comparisons showed a significant gradual increase in the number and intensity of compounds observed that may be caused by the degradation of other molecules in the sample extract. The approach can be applied as preliminary work in a metabolite profiling study, helping to set the appropriate workload to produce confident results.
Highlights
Metabolite profiling refers to the identification and quantitation of low molecular weight molecules that may be found in a particular metabolic pathway using hyphenated analytical techniques such as LC-MS/MS [1,2]
We investigated the stability of samples as part of the initial steps in designing an untargeted metabolite profiling study
The effect of time on the stability of extracted wheat floret samples was investigated over a period of time during analysis
Summary
Metabolite profiling refers to the identification and quantitation of low molecular weight molecules that may be found in a particular metabolic pathway using hyphenated analytical techniques such as LC-MS/MS [1,2]. One of the many analytical technologies used for the study of metabolites is liquid chromatography coupled to high-resolution mass spectrometry, or LC-HR-MS. This analytical technique has been widely used for its capacity to separate and detect a diverse set of molecules with high sensitivity [2,5]. An example of a comprehensive untargeted protocol using an ultra-high-performance LC-MS by De Vos et al [6] resulted in the detection of several hundred metabolites from the analysis of Arabidopsis seedlings. This work shows quality assurance results that ensure the stability of the masses detected for 240 h [6]
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