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This study aims to develop a bio-based wood adhesive with excellent water resistance, focusing on environmentally friendly solutions. The synthesis of an oxidized starch-lignin (OSTL) composite adhesive was accomplished by modifying starch via oxidation and subsequent cross-linking with lignin. Ammonium persulfate (APS) was employed for oxidation of starch, introducing aldehyde groups that upgrade its reactivity with lignin. Subsequently, the oxidized starch (OST) was cross-linked with the phenolic rings of lignin, resulting in a strong network structure. The oxidation of starch and its cross-linking mechanism with lignin were investigated using the Fourier transform infrared (FT-IR), proton nuclear magnetic resonance (1H-NMR), and X-ray photoelectron spectroscopy (XPS) techniques, proving the formation of aldehyde and carboxyl groups with subsequent reaction possibilities. The effects of oxidant dosage, oxidation time, and the ratio of starch to lignin on the adhesive properties were systematically studied. The results demonstrated that the OSTL adhesive, prepared under optimized conditions, exhibited outstanding adhesion strength (1.23 MPa in dry state) and water resistance (0.94 MPa after 24 h cold water immersion, 1.04 MPa after 3 h in hot water, and 0.69 MPa after 3 h in boiling water), significantly outperforming conventional wood adhesives in terms of cold water, hot water, and boiling water resistance. In addition, the thermal behavior of the OSTL adhesive was further validated using differential scanning calorimetry (DSC) as well as thermogravimetric analysis (TGA). This study presents new insights and technical support for the development of green, environmentally friendly, and highly water-resistant lignin-based bio-adhesives.<\/jats:p>","DOI":"10.3390\/polym17111545","type":"journal-article","created":{"date-parts":[[2025,6,2]],"date-time":"2025-06-02T03:19:38Z","timestamp":1748834378000},"page":"1545","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Characterization of Water-Resistant Adhesive Prepared by Cross-Linking Reaction of Oxidized Starch with Lignin"],"prefix":"10.3390","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0009-0005-5939-5697","authenticated-orcid":false,"given":"Chengyuan","family":"Liu","sequence":"first","affiliation":[{"name":"Yunnan Provincial Key Laboratory of Wood Adhesives and Glued Products, College of Material and Chemical Engineering, Southwest Forestry University, Kunming 650224, China"},{"name":"International Joint Research 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