In vivo drug release from solid lipid nanoparticles (SLN) takes place by diffusion and degradation of the lipid matrix. SLN with different degree of crystallinity were prepared to study the effect of crystallinity on the degradation velocity. These SLN were produced by using glycerides with different length of fatty acid chains and known differences in crystallisation velocity (Dynasan 114 and 116), and using stabilisers interfering differently with the crystallisation process of the lipid matrix (cholic acid sodium salt (NaCh), Poloxamer 407 (Plx 407)). NaCh disturbs the crystallisation process, Poloxamer shows little interference. The particles were characterised by photon correlation spectroscopy (PCS) and differential scanning calorimetry (DSC), degradation velocity was determined directly after production and during storage up to 4 weeks under different storage conditions using an especially developed assay based on the NEFA Test kit. After production, SLN with a lower crystallinity matrix (Dynasan 114 and 116, NaCh) degraded faster than higher crystalline particles (all SLN with Plx 407), and showed a decrease in degradation velocity with increasing crystallinity during storage. Fast crystallising particles made from Dynasan 116 stabilised with the non-interfering Plx 407 showed no change in the degradation velocity during storage. SLN produced with a higher crystalline lipid in combination with the crystallisation-disturbing NaCh (Dynasan 116, NaCh) required a 'ripening time' to reach sufficient crystallinity.