Human well-being can be affected by the loss of ecosystem services from conversion of agricultural lands. Uncovering negative ecological consequences of rural-urban conversion is important for regulating rural-urban land conversion. This paper evaluates the impacts of rural-urban land conversion on the ecological well-being of different interest groups in China and makes policy recommendations for mitigating them. This research empirically quantifies and compares changes in the ecological well-being of rural and urban residents due to rural-urban land conversion and examines how transformation factors affect such changes in Hubei, China using the Fuzzy Synthetic Evaluation Model. Results show that compared with urban residents, rural resident ecological well-being level declines more obviously with rural-urban land conversion. Two socio-demographic characteristics, age and education level, as well as zoning characteristics, influence both rural and urban resident well-being changes. It is argued that there is a need for quantitative measurement of agricultural ecosystem services changes and that the construction of ecological compensation policies in areas undergoing rural-urban land conversion is essential for regulating rural-urban land conversion and for maintaining resident ecological well-being.