Abstract   For the past 20 years, the life cycle assessment (LCA) community has sought to integrate impacts on biodiversity into the LCA framework. However, existing impact assessment methods still fail to do so comprehensively because they quantify only a few impacts related to specific species and regions. This paper proposes a methodological framework that will allow LCA practitioners to assess currently missing impacts on biodiversity on a global scale. Building on existing models that seek to quantify the impacts of human activities on biodiversity, the herein proposed methodological framework consists of 2 components: a habitat factor for 14 major habitat types and the impact on the biodiversity status in those major habitat types. The habitat factor is calculated by means of indicators that characterize each habitat. The biodiversity status depends on parameters from impact categories. The impact functions, relating these different parameters to a given response in the biodiversity status, rely on expert judgments. To ensure the applicability for LCA practitioners, the components of the framework can be regionalized on a country scale for which LCA inventory data is more readily available. The weighting factors for the 14 major habitat types range from 0.63 to 1.82. By means of area weighting of the major habitat types in a country, country-specific weighting factors are calculated. In order to demonstrate the main part of the framework, examples of impact functions are given for the categories “freshwater eutrophication” and “freshwater ecotoxicity” in 1 major habitat type. The results confirm suitability of the methodological framework. The major advantages are the framework's user-friendliness, given that data can be used from LCA databases directly, and the complete inclusion of all levels of biodiversity (genetic, species, and ecosystem). It is applicable for the whole world and a wide range of impact categories. Integr Environ Assess Manag 2018;14:282–297. © 2017 SETAC Key Points Impact assessment methods can help to evaluate human made impacts on biodiversity in order to stop the alarming decrease of life on earth. The proposed methodological framework brings life cycle assessment and biodiversity together in order to assess product environmental impacts on biodiversity. A new approach is developed based on existing ideas. This approach can be applied globally and with regard to different human-made impacts.