Abstract: The present invention discloses a quantitative detection method of multiple metabolic components in a biological sample and a metabolic chip used in the method. The detection method includes performing derivatization treatment on the biological sample and then detecting the derivatized biological sample by liquid chromatography-mass spectrometry. During derivatization treatment, 3-nitrophenylhydrazine is used as a derivatization reagent, and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide is used as a derivatization reaction catalyst. According to the detection method of the present invention, high-sensitivity detection can be achieved, multiple metabolic components of different magnitudes can be detected, the operation is simple and fast, and the method is applicable to clinical detection and scientific research examination.