Development and application of physically separated references for aqueous (1)H DOSY diffusion coefficient-formula weight (D-FW) correlation analysis is reported. Commercially available biological buffers (Tris and HEPES) and a water-soluble alcohol (tert-butanol) were used as physically separated references for a Ru and a Mn complex in D(2)O. This extension of DOSY D-FW analysis expands its applicability to a wide variety of water-soluble molecules or metal complexes, with particular application to green chemistry.