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. 2022 Jun 23;7(27):23061-23068.
doi: 10.1021/acsomega.2c00089. eCollection 2022 Jul 12.

Direct Determination of Free Nicotine Content in Tobacco

Li Li  1 , Hongfei Zhang  2 , Jing Wen  1 , Yi Shen  1 , Dongliang Li  1 , Cheng Luo  1 , Jian Zheng  1 , Juan Yang  1
Affiliations

Direct Determination of Free Nicotine Content in Tobacco

Li Li et al. ACS Omega. .

Abstract

The accurate determination of the free nicotine content in tobacco is of great importance for the quality assessment of tobacco leaves and cigarettes, as well as for the study of tar reduction and cigarette formulation. A method based on solvent extraction and gas chromatography-mass spectrometry (GC-MS) was established for direct determination of free nicotine content in tobacco. Free nicotine extraction from tobacco was optimized, using different solvents and physical techniques (shaking, ultrasonic oscillation, and static extraction). Ultimately, a 24 h static extraction with cyclohexane, followed by GC-MS analysis, gave superior results. The standard addition recovery was 98.0-104.7%, with a limit of detection of 5.3 μg/g and a relative standard deviation between 1.3 and 4.1% (n = 5). Quadratic regression of the standard curve was excellent (R 2 ≥ 0.9994). The free nicotine content was determined in 67 tobacco samples, with parallel samples showing relative deviations of 0.1-3.1%. To evaluate the effect of nicotine salts present in tobacco, malic acid, citric acid, acetic acid, and nicotine citrate were spiked into samples prior to extraction and analysis. The results show no interference from bound nicotine compounds on the method. The experimental results show that direct determination of free nicotine in tobacco was achieved using a simple static cyclohexane extraction. Moreover, the high extraction efficiency of free nicotine was also achieved with ease of operation and good repeatability.

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Conflict of interest statement

The authors declare no competing financial interest.

Figures

Figure 1
Figure 1
Schematic of the extraction procedure using cyclohexane. SSP and CY represent 0.1 g of sieved sample powder and 50 mL of cyclohexane, respectively. FR, filter residue; GC–MS, gas chromatography–mass spectrometry; SE, static extraction.
Figure 2
Figure 2
Quadratic regression equations fitted to a standard curve under different experimental conditions. SR50, SR20, and SR10 represent the split ratios of 50:1, 20:1, and 10:1 respectively.
Figure 3
Figure 3
GC–MS analysis chromatogram of the internal standard and sample. (a) Standard solution; (b) sample solution: anethole (1) and nicotine (2). EI, electron ionization; GC–MS, gas chromatography–mass spectrometry; TIC, total ion current; SIM, selected ion monitoring.
Figure 4
Figure 4
Diagram of the plant cell structure.
See this image and copyright information in PMC

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