Fuente: WIPO "tobacco agriculture" The invention discloses a tobacco root metabonomic analysis method based on a gas chromatography-mass spectrometry. The tobacco root metabonomic analysis method comprises the steps of pre-treating a sample, carrying out chromatographic-mass spectrum separation, identifying a metabolite structure, quantitatively analyzing metabolites and analyzing data. The tobacco root metabonomic analysis method takes a polar solvent and a non-polar solvent to extract polar metabolites and non-polar metabolites of tobacco roots respectively, and all the collected metabolites collected by the gas chromatography-mass spectrometry are qualitatively identified through adopting retention indexes and a mass spectrogram library; and all the collected metabolites are quantitatively analyzed by adopting a non-target quantitative analysis method. The establishment of the method provides important references to development of relative researches of tobacco root metabonomics.