Comparison of Analytical Methods in Chemometric Fingerprinting of Metallicolous and Non‐metallicolous Populations of Echium vulgare L

INTRODUCTION: Adverse environmental conditions usually change plant biochemical pathways resulting in accumulation or decreased content of both primary and secondary metabolites. The chemometric fingerprinting analysis proves to be a useful tool to reveal phytochemical differentiation between plants inhabiting heavy metal‐contaminated and uncontaminated areas. OBJECTIVE: Development and assessment of four analytical techniques – high performance capillary electrophoresis (HPCE), thin‐layer chromatography (TLC), mass spectrometry (MS), and Fourier transform infrared (FTIR) spectroscopy in chemometric fingerprinting of metallicolous and non‐metallicolous populations of Echium vulgare L. MATERIAL AND METHODS: Twenty‐one crude methanol extracts of shoot samples representing three populations of Echium vulgare L., two originating from highly metal polluted areas and one from an unpolluted area, were investigated using four analytical methods: HPCE, TLC, MS, and FTIR spectroscopy. Data pre‐processing (denoising, background subtracting, horizontal alignment) followed by principal component analysis (PCA), hierarchical clustering analysis (HCA), and phytochemical difference index (DI) calculations facilitated exploration of the differences and similarities between the populations. RESULTS: Clear phytochemical divergence between metallicolous and non‐metallicolous populations of Echium vulgare was found. The suitability of the analytical techniques for revealing phytochemical markers and discrimination of individuals originating from different populations differed and in general increased in the order: TLC < MS = HPCE < FTIR. CONCLUSION: The chemometric methods applied were successful in discrimination between samples from polluted and unpolluted areas, showing a potential perspective for environmental quality control. Copyright © 2016 John Wiley & Sons, Ltd.