In this paper, novel amide-hydroxamic acid surfactants, such as (N-(6-(hydroxyamino)-6-oxohexyl)benzamide (NHOB), N-(6-(hydroxyamino)-6-oxohexyl) octanamide (NHOO), N-(6-(hydroxyamino)-6-oxohexyl) decanamide (NHOD) and N-(4-(hydroxyamino)-4-oxobutyl) octanamide (NOBO), wereintroduced as flotation collectors for selective separation of scheelite from calicite. The micro-flotationresults demonstrated that compared to NHOO, NOBO and NHOB, NHOD exhibited superior collectingpower to scheelite and enabled the separation of scheelite and calcite under pH around 10. Thecalculation results of density functional theory (DFT) and C log P (octanol–water partition coefficient)indicated that NHOD owned excellent hydrophobicity and affinity to scheelite surfaces. The analyses ofFTIR spectra, zeta potential and XPS illustrated that besides the electrostatic attraction with Ca cationicspecies on scheelite surfaces, NHOD might chemisorb onto scheelite by formation of NHOD-W surfacecomplexes, resulting in a superior selectivity to scheelite compared with calcite. NHOD’s uniqueproperties, such as characteristic bond patterns on scheelite surfaces, double hydrophobic groups, andintermolecular hydrogen bonds between neighboring molecules coated on scheelite surfaces, renderedit to be a superior collector for scheelite/calcite flotation separation.