Tick hemolymph plays an important role in the transportation of nutrients as well as metabolites. The hemolymph consists of plasma and blood cells, and proteins are the main components of plasma. This study aimed to investigate the protein composition of the hemolymph of Haemaphysalis qinghaiensis and to explore the effects of different hosts on the hemolymph protein composition of ticks, which could provide a reference for the screening of tick-protective antigens. Hemolymph was collected from the engorged females of the H. qinghaiensis ticks from the Bos grunniens (HqB) and Ovis aries (HqO) hosts. We identified 17 host-derived high-confidence proteins and 156 tick-derived high-confidence proteins from HqB. Fifteen host-derived high-confidence proteins and 155 tick-derived high-confidence proteins were identified from HqO. There were significant differences in the composition and abundance of the host-derived protein in the hemolymph from the two sources, with fibrinogen, alpha-1-antiproteinase, α-2-macroglobulin, and an uncharacterized protein present only in HqB, while ubiquitin-60S ribosomal protein L 40 was found only at HqO. Besides, the abundance of these proteins also varied significantly. The 163 tick-derived proteins identified are classified as enzymes, inhibitors, transporters, immunity-related proteins, cytoskeletal proteins, heat shock proteins, nuclear proteins, other proteins, uncharacterized proteins, and secreted proteins. The KGD sequence of A1 in the uncharacterized protein suggested that the unidentified protein may be associated with anti-coagulants, but further research was needed to confirm the function of these uncovered proteins. HqB and HqO shared a total of 148 tick-derived proteins, with eight proteins present only in HqB and seven only in HqO. The abundance of 65 shared proteins was significantly higher in HqO. In conclusion, the hemolymph proteins of H. qinghaiensis tick were composed of host-derived and tick-derived proteins. Different blood meals had a large effect on the composition and abundance of both host-derived and tick-derived proteins.