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A novel method has been developed for the rapid extraction and determination of 8 defoliations (including thidiazuron, butiphos, methabenzthiazuron, abscisic acid, carfentrazone-ethyl, diuron, paraquat, and pyrithiobac-sodium) in cotton by accelerated solvent extraction (ASE) coupled with ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS).