The Jinying gold deposit is located in southern Jilin Province in northeast China and is representative of the large Early Cretaceous gold deposits in this area. To better understand ore genesis of this deposit, a multi-isotope integrated analysis of U–Pb–Rb–Sr–He–Ar–S has been carried out. Laser ablation inductively coupled plasma–mass spectrometry (LA–ICP–MS) dating of zircons from the granodiorite porphyry and dioritic porphyrite in the study area yields ages of 172.1 ± 1.2 Ma and 122.5 ± 0.8 Ma, suggesting that corresponding intrusion occurred in the Middle Jurassic and the Early Cretaceous. Rb–Sr dating of the pyrite yields an isochron age of 120 ± 3 Ma, suggesting that gold mineralization occurred in the Early Cretaceous. The fluid inclusions in pyrite yield 3He/4He ratios clustered within a small range from 0.08 to 0.13 Ra, 40Ar/36Ar ratios between 331.6 and 351.3, and mantle He in the range of 1.0–1.6%, indicating that the ore-forming fluids originated from a mixed crustal and mantle source. The in situ S isotopic values of pyrite vary between + 0.1 ‰ and + 2.8 ‰, suggesting that the ore-related sulphur came from the deep magmatic source. Combined with the geological history of the study area, it can be concluded that the gold mineralization was possibly related to the extensional setting associated with the rollback of the Palaeo-Pacific Plate.