In this paper we present the development of a method for the detection of toxic substances on ancient arrow points. The aim is to go back in time until the Palaeolithic period in order to determine if poisonous substances were used to enhance the hunting weapons. The ethnographic documentation demonstrates that hunters of every latitude poisoned their weapons with toxic substances derived from plants and occasionally from animals. This highlights that often the weapons would be rather ineffective if the tips were not poisoned. The fact that toxic substances were available and the benefits arising from their application on throwing weapons, suggests that this practice could be widespread also among prehistoric hunters. The project reviewed the research of the toxic molecules starting from current information on modern plants and working backwards through the ages with the study of ethnographic and historical weapons. This knowledge was then applied to the archaeological material collected from International museum collections. Results have shown that using this method it is possible to detect traces of toxic molecules with mass spectrometry (MS) and hyphenated chromatographic techniques even on samples older than one hundred years, which we consider a positive incentive to continue studying plant poisons on ancient hunting tools.