Third-hand smoke (THS) is composed of surface-deposited remnants resulting from tabacco-smoking. Because THS components have properties of remaining on, re-emitting from and reacting on and with surfaces, in-situ analysis of the components on different surfaces is both in high demand and challenging. The aim of this study is to establish desorption corona beam ionization (DCBI)-MS/MS as an analytical tool for THS research. To this end, an in-situ DCBI-MS/MS approach was developed for the quantitative analysis of typical THS environmental markers, i.e. nicotine and cotinine on different surfaces such as fruits, cotton clothing, glass, and toys etc. The limits of detection of nicotine and cotinine were both 1.4 μg m-2. Low-temperature DCBI-MS/MS was applied to the direct detection of THS on fingers without any skin damage. Smoking-related biomarkers analyses in urine were accomplished, with a 10 s DCBI analysis time. The on-surface tobacco-specific nitrosamines (TSNAs), such as 1-(N-methyl-N-nitrosamino)-1-(3-pyridinyl)-4-butanal) (NNA), 4-(methylnitrosamino)-1-(3-pyridinyl)-1-butanone (NNK), and N-nitroso nornicotine (NNN) were in-situ successfully detected in dust samples.