Dissolved organic carbon (DOC) from Oa horizons has been proposed to be an important contributor for subsoil organic carbon stocks. We investigated the fate of DOC by directly injecting a DOC solution from 13C labelled litter into three soil depths at beech forest sites. Fate of injected DOC was quantified with deep drilling soil cores down to 2 m depth, 3 and 17 months after the injection. 27 ± 26% of the injected DOC was retained after 3 months and 17 ± 22% after 17 months. Retained DOC was to 70% found in the first 10 cm below the injection depth and on average higher in the topsoil than in the subsoil. After 17 months DOC in the topsoil was largely lost (− 19%) while DOC in the subsoil did not change much (− 4.4%). Data indicated a high stabilisation of injected DOC in the subsoils with no differences between the sites. Potential mineralisation as revealed by incubation experiments however, was not different between DOC injected in topsoil or subsoils underlining the importance of environmental factors in the subsoil for DOC stabilisation compared to topsoil. We conclude that stability of DOC in subsoil is primary driven by its spatial inaccessibility for microorganisms after matrix flow while site specific properties did not significantly affect stabilisation. Instead, a more fine-textured site promotes the vertical transport of DOC due to a higher abundance of preferential flow paths. © 2020, The Author(s).