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There is an increasing expectation on the private sector to address biodiversity impacts and contribute towards global conservation goals. Appropriate evidence use can help businesses avoid biodiversity losses and realise gains, reduce resources wasted on ineffective or suboptimal action, whilst minimising biodiversity-related risks and securing opportunities from engaging with biodiversity. We review the status of evidence-based action in the private sector, where previous studies have identified concerning trends, and explore the barriers that may currently be hindering evidence use. To learn from this, and improve evidence use, we propose a set of principles for evidence-based biodiversity impact mitigation. We outline tools and resources that can help businesses move towards evidence-based practice and achieve each of these principles. Meeting these principles would improve the biodiversity outcomes from business’ biodiversity related actions. However, for business action to contribute more fully to global conservation goals, broader political and socio-economic issues also need addressing.