Haemaphysalis longicornis is a parasitic arthropod on the body surface of vertebrates, which lives by sucking host blood. In addition, H. longicornis can carry and transmit a variety of deadly pathogens to the host. The control of ticks mainly depends on chemical insecticides, among which chlorpyrifos is a common drug to control them. The salivary gland is an important organ to promote the blood-sucking of ticks and regulate the development of ticks, meanwhile, it plays an important role in protecting against pesticides. With the increasing drug resistance of ticks to chlorpyrifos and the harm of the drug to the environment, it is urgent to find other drugs with low toxicity and high efficiency to replace chlorpyrifos. In this study, DIA quantitative proteomics was used to study the dynamic changes of all proteins in the salivary gland of H. longicornis treated with chlorpyrifos. Bioinformatics analysis was used to analyze these proteins, and a variety of proteins that may be involved in the detoxification process were found, such as glutathione S-transferase, glutamine synthase and a variety of heat shock proteins, which provided new molecular targets and ideas for the development of anti-tick vaccines and related drugs.