US 12,131,893 B1
Method of identifying tobacco carbonyl components using non-targeted mass spectrometry
Ruizhi Zhu, Kunming (CN); Xiaoxi Si, Kunming (CN); Yanqun Xu, Kunming (CN); Zhihua Liu, Kunming (CN); Ji Yang, Kunming (CN); Wei Jiang, Kunming (CN); Chunbo Liu, Kunming (CN); Fengmei Zhang, Kunming (CN); Zhenjie Li, Kunming (CN); and Shiyun Tang, Kunming (CN)
Assigned to CHINA TOBACCO YUNNAN INDUSTRIAL CO., LTD., Kunming (CN)
Filed by CHINA TOBACCO YUNNAN INDUSTRIAL CO., LTD., Kunming (CN)
Filed on Oct. 30, 2023, as Appl. No. 18/497,574.
Application 18/497,574 is a continuation of application No. PCT/CN2023/102744, filed on Jun. 27, 2023.
Claims priority of application No. 202310751411.3 (CN), filed on Jun. 25, 2023.
Int. Cl. G01N 30/86 (2006.01); G01N 1/10 (2006.01); G16C 20/20 (2019.01); H01J 49/00 (2006.01); H01J 49/42 (2006.01); G01N 30/02 (2006.01); G16C 20/30 (2019.01)
CPC H01J 49/0036 (2013.01) [G01N 1/10 (2013.01); G01N 30/8631 (2013.01); G01N 30/8679 (2013.01); H01J 49/425 (2013.01); G01N 2001/1006 (2013.01); G01N 2030/027 (2013.01); G16C 20/20 (2019.02); G16C 20/30 (2019.02)] 20 Claims
OG exemplary drawing
 
1. A non-targeted mass spectrometry identification method for identifying tobacco carbonyl components, comprising:
soaking a tobacco product with water;
oscillation extracting a tobacco extract from the tobacco product with acetonitrile;
adding 2,4-dinitrophenylhydrazine (DNPH) to a first sample of the tobacco extract and derivatizing the first sample of the tobacco extract with the DNPH to obtain a first derivatized tobacco sample;
adding DNPH-d3 to a second sample of the tobacco extract and derivatizing the second sample of the tobacco extract with the DNPH-d3 to obtain a second derivatized tobacco sample with an isotope-label;
mixing the first derivatized tobacco sample and the second derivatized tobacco sample in a ratio of 1:1 thoroughly, and filtering the first and second derivatized tobacco samples through a filter membrane to obtain a third derivatized tobacco sample;
preparing a blank sample by oscillation extracting a water extract from water with acetonitrile, derivatizing a first sample of the water extract with DNPH to obtain a first derivatized water sample, derivatizing a second sample of the water extract with DNPH-d3 to obtain a second derivatized water sample, mixing the first derivatized water sample and the second derivatized water sample in a ratio of 1:1 thoroughly, and filtering the first and second derivatized water samples through a filter membrane to obtain the blank sample;
performing ultra performance liquid chromatography (UPLC) and ion trap high resolution mass spectrometry (HRMS) on the first derivatized tobacco sample, the second derivatized tobacco sample, the third derivatized tobacco sample and the blank sample to obtain original LC-MS data;
performing peak detection, peak alignment and peak grouping on the original LC-MS data to obtain original mass spectral feature data of coexisting target components and interfering components, including m/z values, retention times and peak intensities of chromatographic peaks;
filtering the original mass spectral feature data based on statistical characteristics, mass loss, paired chromatographic peaks based on DNPH and DNPH-d3 derivatization markers, and either secondary mass spectrometry or secondary mass spectrometry multiple ion mass spectrum characteristics to obtain final retained chromatographic peaks; and
annotating the final retained chromatographic peaks and structurally identifying the tobacco carbonyl components using the final retained chromatographic peaks.