The information on molecular weight (MW) characteristics of DOM and relevant evolution behaviors during composting are limited. In this study, DOM extracted from co-composting of chicken manure and rice husks were comprehensively analyzed by using high-performance size exclusion chromatography (HPSEC) combined with a two-dimensional correlation spectroscopy (2D COS) to explore the evolution characteristics of MW of compost DOM. The HPSEC detected at UV of 254 nm and at uorescence (FL) Ex/Em wavelengths (315/410, 270/455 nm) all showed a gradual increase in both weight-average and number-average MW for DOM, suggesting that the large MW fractions were continuously generated and polymerized during composting. The 2D COS applied on HPSEC-UV and -FL further identi ed the key active MW chromophoric (i.e., 0.5, 7.2. 9.5, 26.3, 30.7, and 83.9 kDa) and uorophoric (i.e., 0.55 and 3.5 kDa) molecules that mainly participated in the transformation processes of compost DOM. Moreover, these active MW species were preferentially formed by the order of small to large molecules. A hetero-2D COS analysis disclosed the change sequence in the order of 0.5 and 7.2 kDa chromophores → 3.5 kDa uorophores, and the 0.55 and 3.5 kDa uorophores → 26.3 and 83.9 kDa chromophores.