Human remains from excavation sites in southern China, dated from 100 to 10,000 years, were investigated for nicotine. The nicotine concentrations were measured by radioimmunoassay. Consequently, the values measured represent the sum of nicotine and its metabolites. Positive results were observed in 25.9% (mean values of all samples). The concentrations ranged from 11.8 to 474.2 ng/g sample. From the excavation sites with the highest number of individuals, Chongtong (18 individuals) were 38.9%, and from the site Leigund (21 individuals) were 28.6% from the samples nicotine positive. The highest level with 474.2 ng/g nicotine was found in a sample dated 100 years. It is possible, that this individual was smoker. In addition the samples were investigated also by gas chromatography/mass spectrometry. The GC/MS indicated the presence of nicotine and its primary metabolite, cotinine. The presence of cotinine points out the assumption that nicotine was used antemortem. The detection of unchanged nicotine indicates that nicotine remained present in bones up centuries.