The aim of this study was to evaluate the adhesive interface with dentin of an experimental adhesive resin with nanostructured hydroxyapatite addition. The organic phase of the adhesive resin was produced by mixing 50 wt.% Bis-GMA, 25 wt.% TEGDMA and 25 wt.% HEMA. CQ and EDAB were added at 1 mol% to all groups, according to the monomer moles. HAnano was added at the following two concentrations: 0 and 2 wt%. One commercial adhesive system was used as control. Nine lower incisor bovine teeth were used to produce interfaces of adhesive resin and dentin. Tooth slices were analysed using the following micro-Raman parameters: a 100 mW diode laser with 785 nm wavelength and spectral resolution of ~ 3–5 cm-1. One-dimensional mapping was performed over a 150 μm line across the adhesive-dentine interface at 1 μm intervals using a computerized XYZ stage. These areas covered the composite resin, adhesive layer, hybrid layer, partially demineralised and un-affected dentine and were visualised and focused at x500 magnification. Raman analysis showed the penetration of experimental and commercial adhesive systems into dentin. HAnano was observed into the hybrid layer. Based on results of the present study, is possible to observe resin and nanostructured hydroxyapatite penetration at the hybrid layer.