Benzidine and dimethyl yellow reagents have been used for distinguishing heartwood from sapwood in the Pinaceae and Araucariacae families, and in Eucalypt species. Both have been classified as carcinogenic by European and United States authoritites, yet the need for effective heartwood/sapwood differentiation remains, not only in the laboratory but also out in the timber processing chain. Safer alternative reagents have been proposed over the past twenty years, but some of these have since been linked with health problems, and most of the rest lack the specificity and general usefulness of those traditionally used. Recent research has investigated various azo, diazo, and other nitrated reagents. Safe but useful alternative methods of sapwood/heartwood differentiation are being established, and recommendations are made for several timber species.

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