The invention provides a solid-phase extraction column and a preparation method thereof. At the bottom of a column tube, the following operations are carried out in sequence: an outlet sieve plate is pressed in, a graphitized carbon black layer is filled in, a layer of insulating sieve plate is pressed in, a multi-amination packing layer is filled in, a hydrophobic group bonded silica gel resin layer is filled in after being pressed into the insulating sieve plate, and an inlet sieve plate is finally pressed in; and an outlet of the extraction column is arranged at the bottom of the column tube. The solid-phase extraction column provided by the invention is a pesticide residue detection special column and is made of a three-layer adsorbing material. Impurities, such as amino acid microelements, and heavy metal, can be removed by selectively adsorbing and selectively eluting and purifying a matrix of a complex sample, so that the defect of poor purification effect of the conventional solid-phase extraction product can be avoided, the cleaning degree of the sample is greatly improved, an analyzer is effectively protected, the accuracy of sample detection is improved, the detection sensitivity is improved, the time is saved, and the working efficiency of detectors can be improved. Furthermore, the solid-phase extraction column has the advantages of generality, and low selectivity of a matrix of the sample to be detected, and can be applicable to detection of most food samples.