Abstract: In order to rapidly identify nicotine converters in tobacco plants, a screening method based on gas chromatography-mass spectrometry was developed, and two strategies (with and without incubation) were proposed. The non-incubation strategy took about 5-6 min from sampling to analysis. With this strategy, the complete removal of converters could be achieved with the sacrifice of 9% non-converters. For the incubation strategy, in-tube samples were cured at 37 ℃ for 12 h before the extraction and analysis. The complete separation of converters from non-converters in 221 seedlings was achieved by using the incubation strategy. The developed method provides a technical support for the fast and efficient identification of nicotine converters from tobacco seedlings.