title: The reproducibility of liquid chromatography separation technology and its potential impact on large scale plant metabolomics experiments.
abstract: Unraveling the constituents of biological samples using HPLC is a central core technology in metabolomics experiments. Consistency in retention time across many samples is a critical criterion for judging the quality of a data set, which must be met before further analysis are possible. Here, the performance of two ultra high-performance liquid chromatography (UHPLC) systems has been compared using an established separation protocol optimized for phenylpropanoids, a class of secondary compounds found in plants displaying intermediate polarity. The two systems differed markedly with respect to their reproducibility and pressure stability. The standard deviation of the retention time of representative peaks differs up to 30-folds between the systems. Adjustments made to the gradient profiles succeeded in equalizing their level of performance. However, the modifications made to the separation protocol reduced the quality of the separation, particularly of the more rapidly eluting components, and lengthened the run time.
authors.affiliation: Leibniz Institute of Plant Genetics and Crop Plant Research IPK, Correnstraße 3, 06466 Gatersleben, Germany.
authors.forename: Nadja
authors.initials: N
authors.lastname: Arens
authors.affiliation: Leibniz Institute of Plant Genetics and Crop Plant Research IPK, Correnstraße 3, 06466 Gatersleben, Germany.
authors.forename: Stefanie
authors.initials: S
authors.lastname: Döll
authors.affiliation: Leibniz Institute of Plant Genetics and Crop Plant Research IPK, Correnstraße 3, 06466 Gatersleben, Germany. Electronic address: mock@ipk-gatersleben.de.
authors.forename: Hans-Peter
authors.initials: HP
authors.lastname: Mock
mesh_terms: D027461:Beta vulgaris / Q000737:chemistry / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D055442:Metabolome; D055432:Metabolomics / Q000379:methods*; D018515:Plant Leaves / Q000737:chemistry / Q000378:metabolism; D015203:Reproducibility of Results
keywords: MS based detection; Metabolomic profiling; Retention time; Secondary metabolites; U(HPLC)