Gold nanoparticles (AuNPs) assisted laser desorption ionization mass spectrometry (LDI-MS) emerged as an effective technique for the detection of analytes with high sensitivity. The surface chemistry and the size of AuNPs are the crucial parameters for lowering the detection limits and increasing the selectivity of LDI-MS. Here we show that chemical-free size selected AuNPs, obtained by laser ablation synthesis in solution (LASiS), have very low background in the low mass region (<500 Da), contrary to citrate stabilized AuNPs (citrate-AuNPs) and dihydroxyacetophenone (DHAP). This allowed better performances for the picomole detection of low mass analytes like arginine, fructose, atrazine, anthracene and paclitaxel. The results suggest that chemical-free LASiS-AuNPs can be an excellent matrix for nanoparticle-assisted LDI-MS.