Yamuna, a prominent river of India covers an extensive area of 345,843 km2 from Yamunotri glacier through six Indian states. Residues of organochlorine pesticides (OCPs) namely, isomers of HCH and endosulfan, DDT and its metabolites, aldrin, dieldrin, were analysed in water of river Yamuna along its 346 km stretch passing through Haryana–Delhi–Haryana and the canals originating from it. β-HCH, p.p′-DDT, p.p′-DDE and p.p′-DDD had maximum traceability in test samples (95–100%) followed by γ-HCH, α-HCH and o.p′-DDD (60–84%) and o.p′-DDT, δ-HCH and o.p′-DDE (7–30%) while aldrin, dieldrin, α and β endosulfan remained below detection limits (BDL). The concentration of ΣHCH and ΣDDT at different sites of the river ranged between 12.76–593.49 ng/l (with a mean of 310.25 ng/l) and 66.17–722.94 ng/l (with a mean of 387.9 ng/l), respectively. In canals the values were found between 12.38–571.98 ng/l and 109.12–1572.22 ng/l for ΣHCH and ΣDDT, respectively. Water of Gurgaon canal and Western Yamuna canal contained maximum and minimum concentration, respectively both of ΣHCH and ΣDDT residues. Sources of these pesticides and suggested measures to check pesticide pollution of this major Indian river, keeping in view its vital link with life, are discussed in this paper.