The invention provides a pretreatment method of detection on malachite green, crystal violet and residues of malachite green and crystal violet in eels, belonging to the field of analytical chemistry. The method comprises the operations of weighing of a sample to be detected, homogenization, solid phase extraction, elution and the like; instruments used in the method comprise a constant temperature oscillator, a high speed dispersing homogenizer, a centrifugal machine and the like. A sample solution prepared by using the pretreatment method can be directly detected by using a liquid chromatogram-tandem mass spectrometry method; compared with the prior art, the pretreatment method has the advantages of simple and effective operation, less time consumption, low cost, good reproducibility and the like, and has very strong operability, so that the pretreatment method has the hope to be widely popularized and applied in a large scale in industries such as environment protection, commodity inspection, entry and exit inspection and quarantine and the like, with significant economic benefits.