Swarming honey bee (Apis mellifera L.) colonies rear supernumerary young queens that compete for the limited resources (workers) necessary for founding a new colony. Young queens often fight to death. Queens show several adaptations to fight and conflict, such as short developmental time and early onset of venom production. During fights, queens often release rectal fluid with a strong smell of wine grapes, after which they temporarily stop fighting. This potentially reduces individual overall risk of deadly injury. The fluid and one of its components, ortho-aminoacetophenone, were previously found to have a pheromonal effect on workers, but the evidence is equivocal. Recently, it has been suggested that the effects of this substance may be context- or concentration-specific. We performed semi-quantitative gas-chromatography mass spectrometry (GC/MS) analysis of the fluid (1) released by queens during their first fight, (2) released during a subsequent fight, and (3) obtained by dissecting the hindgut of queens and (4) of workers. Following preliminary results by Page et al. 1988 (Experientia 44: 270-271), we scored presence / absence of eight substances. Five substances (ortho-aminoacetophenone, decanoic acid, dodecanoic acid, octyl decanoate, and decyl decanoate) were characteristic of queens only. ortho-Aminoacetophenone was detected in all queen and in none of the worker samples, in agreement with previous findings that worker faeces do not have any pheromonal effect. The fluid released by queens on their second fight also contained ortho-aminoacetophenone, but in smaller quantities. These data substantiate previous bioassay results, and provide estimates of ortho-aminoacetophenone concentration as required to design experiments addressing the function and adaptive significance of this behaviour.