The invention relates to a novel Avermectin derivative and a preparation method thereof. The preparation method comprises the following steps: A, dissolving Avermectin B1a or Avermectin B2a into an organic solvent; B, cooling down the solution, adding a substitution reagent and dripping an acid-capturer; C, adding the reaction solution into a sulfuric acid solution, regulating pH, stilling, layering, extracting, adding an organic phase into an ammonia solution, regulating pH to be neutral, stilling, layering, extracting and merging the organic phase; D, performing distillation of the organic phase under reduced pressure to obtain the product. According to the invention, on the basis of the Avermectin B2a or B1a, C5-OH is subject to further chemical modification, and compared with the common Avermectin product, the novel Avermectin derivative has the advantages of high drug effect, broad insecticidal spectrum, good persistence, no residues, no pollution, high efficiency and no resistance. The novel Avermectin derivative has extremely high activity on many pests and mite, has the function of not only stomach toxicity but also killing and has a very good insecticide effect at a very low dosage.