Magnetic solid phase extraction integrated with in situ derivations for the profiling of 12 phytohormones in a single rapeseed seed was developed by using ultra-high performance liquid chromatography-tandem mass spectrometry. The Fe3O4@Ti3C2@β-cyclodextrin nanoparticles were firstly synthesized and used as an adsorbent for the solid-phase extraction of phytohormones. The magnetic dispersive solid-phase extraction and in situ derivation by the addition of N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide were ingeniously combined. This efficient pre-treatment method integrated the extraction, purification, and derivatization processes into one single step. Satisfactory methodological performance was achieved by optimization of the parameters. Linearities (R2 > 0.9928) and recoveries (80.4 %-115.1%) at three spiked levels, as well as the low matrix effect (from -16.63% to 17.06%) and limits of detection (0.89-13.62 pg/mL) were obtained. The spatio-temporal profiling of target phytohormones in different tissues of rapeseed germination was investigated. This method was successfully employed for analyzing target phytohormones in different oilseeds samples.