Spectrophotometric and Chromatographic Determination of Alkaloids and Nicotine Contents in Lebanese Tobacco Leaves
Year 2024,
, 335 - 345, 30.06.2024
Sylvana Elchamieh
,
Ali Jaber
,
Ghassan Ibrahim
,
Edmond Cheble
Abstract
Nicotiana Tabacum; the annual herb; known as Tobacco from the Solanaceae family was known for its alkaloids and especially nicotine (NCT) content; smoking products, insecticides, anthelminthic activity and clinically proven therapeutics are examples of its uses. Herein two different methods were applied in an attempt to quantify the total alkaloids and NCT content in Nicotiana tabacum cultivated in Lebanon. Total alkaloids were investigated through the formation of a complex with bromocresol green under a simple spectrophotometric method. Whilst HPLC-DAD was the choice for the quantitation of NCT levels. The column was Lichrospher select B (5 µm, 250x4 mm), the temperature was set at 29 ⁰C and the wavelength at 260 nm. The mobile phase consisted of 2 M O-phosphoric acid and methanol (60:40, v/v) using isocratic elution at 1 mL/min. A linear relationship was proved under both instruments. The extraction yield of alkaloid totum ranges between 2.1 ± 0.25 and 6.8% ± 0.58 and alkaloids contents range from 12.14 ± 2.01 to 53.12 ± 4.54 mg of AE/g of extract for Ghandouriyeh and Danniye samples respectively. On the other side among the different areas which cultivated Tobacco in Lebanon, Danniye was found to have the highest NCT concentration of dry weight (2.64%) while Al-Hissa possesses the lowest content (0.75%). Even if the results were generally similar to other countries, the study showed a difference in values from one region to another.
References
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- Murray, J. (2014). Nicotine and what Else?: HPLC Elution Optimization for the Analysis of Alkaloids Found in Electronic Cigarettes. University of Tennessee at Chattanooga, United States.
- Perfetti, T. A., Dube, M. F., & Coleman, W. M. (2022). Qualitative and Quantitative Analysis of Nicotine, Nicotine Derivatives, and Nicotine-Related Alkaloid Optical Isomers: A Review. Contributions to Tobacco & Nicotine Research, 31, 175–185. https://doi.org/10.2478/cttr-2022-0018
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- Shikanga, E. A., Kamatou, G. P. P., Chen, W., Combrinck, S., & Viljoen, A. M. (2012). Validated RP-UHPLC PDA and GC–MS methods for the analysis of psychoactive alkaloids in Sceletium tortuosum. South African Journal of Botany, 82, 99–107. https://doi.org/10.1016/j.sajb.2012.05.004
- Shoji, T., Kajikawa, M., & Hashimoto, T. (2010). Clustered Transcription Factor Genes Regulate Nicotine Biosynthesis in Tobacco. The Plant Cell, 22, 3390–3409. https://doi.org/10.1105/tpc.110.078543
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- Tayoub, G., Sulaiman, H., & Alorfi, M. (2016). Determination of nicotine levels in the leaves of some Nicotiana tabacum varieties cultivated in Syria. Herba Polonica, 61, 23–30. https://doi.org/10.1515/hepo-2015-0028
- Tengs, T. O., Ahmad, S., Savage, J. M., Moore, R., & Gage, E. (2005). The AMA proposal to mandate nicotine reduction in cigarettes: A simulation of the population health impacts. Preventive Medicine, 40, 170–180. https://doi.org/10.1016/j.ypmed.2004.05.017
- Tepecik, M., & Ongun, A.R. (2020). Determination of some quality parameters of oriental type tobacco based on harvest times. Turkish Journal of Agricultural Research, 7(2), 156-162. https://doi.org/10.19159/tutad.691727
- Tiwari, R. K., Sharma, V., Pandey, R. K., & Shukla, S. S. (2020). Nicotine Addiction: Neurobiology and Mechanism. 23, 1–7. https://doi.org/10.3831/KPI.2020.23.001
- Wang, S. S., Shi, Q. M., Li, W. Q., Niu, J. F., Li, C. J., & Zhang, F. S. (2008). Nicotine concentration in leaves of flue-cured tobacco plants as affected by removal of the shoot apex and lateral buds. Journal of Integrative Plant Biology, 50, 958-964. https://doi.org/10.1111/j.1744-7909.2008.00684.x
- Yang, S. S., Smetena, I., & Huang, C. B. (2002). Determination of tobacco alkaloids by gas chromatography with nitrogen-phosphorus detection. Analytical and Bioanalytical Chemistry, 373, 839–843. https://doi.org/10.1007/s00216-002-1380-1
- Yasinok, A. E., Sahin, F. I., Eyidogan, F., Kuru, M., & Haberal, M. (2009). Grafting tomato plant on tobacco plant and its effect on tomato plant yield and nicotine content. Journal of the Science of Food and Agriculture, 89, 1122–1128. https://doi.org/10.1002/jsfa.3555
- Zahreddine, Z., Jaber, A., Haidar, S. A., Hosri, C., Ibrahim, G., & Cheble, E. (2021). HPLC-DAD multi-residue method for determination of florfenicol, penicillin and tetracycline residues in raw cow milk. Journal of Clinical and Laboratory Research, 2, 1–8.
- Zhang, J., Ji, H., Sun, S., Mao, D., Liu, H., & Guo, Y. (2007). Selective Determination of Pyridine Alkaloids in Tobacco by PFTBA Ions/Analyte Molecule Reaction Ionization Ion Trap Mass Spectrometry. Journal of the American Society for Mass Spectrometry, 18, 1774–1782. https://doi.org/10.1016/j.jasms.2007.07.017
Year 2024,
, 335 - 345, 30.06.2024
Sylvana Elchamieh
,
Ali Jaber
,
Ghassan Ibrahim
,
Edmond Cheble
References
- Ajanal, M., Gundkalle, M., & Nayak, S. (2012). Estimation of total alkaloid in Chitrakadivati by UV-Spectrophotometer. Ancient Science of Life, 31, 198-201. https://doi.org/10.4103/0257-7941.107361
- Baldwin, I. T., Oesch, R. C., Merhige, P. M., & Hayes, K. (1993). Damage-induced root nitrogen metabolism in Nicotiana sylvestris: Testing C/N predictions for alkaloid production. Journal of Chemical Ecology, 19, 3029–3043. https://doi.org/10.1007/BF00980600
- Beldar, V., Laddha, K. S., Dudure, R. H., Fayed, M. A. A., & Jadhao, M. (2022). An approach for developing a simple and quick method for separation of asiatic acid and asiaticoside rich fraction From Centella Asiatica and simultaneous determination by reversed-phase high-performance liquid chromatography. Yuzuncu Yil University Journal of Agricultural Sciences, 33, 18-28. https://doi.org/10.29133/yyutbd.1175779
- Bhatia, S., Sharma, K., Dahiya, R., & Bera, T. (2015). Modern Applications of Plant Biotechnology in Pharmaceutical Sciences, Oxford, UK: Elsevier Inc.
- Burner, N., McCauley, A., Pramod, S., Frederick, J., Steede, T., Kernodle, S. P., & Lewis, R. S. (2022). Analyses of diverse low alkaloid tobacco germplasm identify naturally occurring nucleotide variability contributing to reduced leaf nicotine accumulation. Molecular Breeding : New Strategies in Plant Improvement, 42, 1-15. https://doi.org/10.1007/s11032-021-01274-5
- Clemens, K. J., Caillé, S., Stinus, L., & Cador, M. (2009). The addition of five minor tobacco alkaloids increases nicotine-induced hyperactivity, sensitization, and intravenous self-administration in rats. International Journal of Neuropsychopharmacology, 12, 1355–1366. https://doi.org/10.1017/S1461145709000273
- Fazel, S., Hamidreza, M., Rouhollah, G., & Verdian-Rizi, M. (2010). Spectrophotometric determination of total alkaloids in some Iranian medicinal plants. Journal of Applied Horticulture, 32, 17-20. https://doi.org/10.37855/jah.2010.v12i01.15
- Food and Drug Administration. (2018). Tobacco Product Standard for Nicotine Level of Combusted Cigarettes Access date: 03.08.2022. https://www.govinfo.gov/content/pkg/FR-2018-03-16/pdf/2018-05345.pdf
- Ganapathi, T. R., Suprasanna, P., Rao, P. S., & Bapat, V. A. (2004). Tobacco (Nicotiana tabacum L.)-A model system for tissue culture interventions and genetic engineering. Indian Journal of Biotechnology, 3, 171-184.
- Goettel, M., Niessner, R., Mueller, D., Scherer, M., Scherer, G., & Pluym, N. (2017). Metabolomic Fingerprinting in Various Body Fluids of a Diet-Controlled Clinical Smoking Cessation Study Using a Validated GC-TOF-MS Metabolomics Platform. Journal of Proteome Research, 16, 3491–3503. https://doi.org/10.1021/acs.jproteome.7b00128
- Gonzalez-Coloma, A., Reina, M., Diaz, C. E., & Fraga, B. M. (2010). Natural product-based biopesticides for insect control. In Comprehensive Natural Products II: Chemistry and Biology. https://doi.org/10.1016/b978-008045382-8.00074-5
- Habib, M. A., Islam, M. M., Islam, M. M., Hasan, M. M., & Baek, K.-H. (2023). Current Status and De Novo Synthesis of Anti-Tumor Alkaloids in Nicotiana. Metabolites, 13, 623. https://doi.org/10.3390/metabo13050623
- Jaber, A. (2017). Matrices MALDI bithiophéniques spécifiques aux alcaloïdes: Étude des mécanismes fondamentaux et applications. Université d’Angers, France.
- Jaber, A., Cheble, E., Abdel Sater, F., Soukariyeh, R., & Khalil, A. (2020). Biological activities of total oligomeric flavonoids enriched extracts of Nicotiana tabacum from eight lebanese regions. Int. J. Pharm. Sci. Rev. Res., 61, 70–77.
- Jaber, A., Hajjar, S., El Riachi, M., Abdel Sater, F., & Cheble, E. (2022). GC-MS Analysis of essential oil and anticancer activities of extracts from discarded leaves of Nicotiana Tabacum Linn. 46, 291–307. https://doi.org/10.33483/jfpau.991289
- Jacob, P., Chan, L., Cheung, P., Bello, K., Yu, L., St Helen, G., & Benowitz, N. L. (2022). Minor Tobacco Alkaloids as Biomarkers to Distinguish Combusted Tobacco Use From Electronic Nicotine Delivery Systems Use. Two New Analytical Methods. Front. Chem., 10. https://doi.org/10.3389/fchem.2022.749089
- John, B., C T, S., George, S., & Reddy, V. R. K. (2014). Spectrophotometric estimation of total alkaloids in selected justicia species. International Journal of Pharmacy and Pharmaceutical Sciences, 6, 647–648.
- Khoi, A. A. M. E. (1983). Spectrophotometric promethazine hydrochloride determination using bromcresol green. Journal of Pharmaceutical Sciences, 72, 704–705. https://doi.org/10.1002/jps.2600720629
- Koetz, M., Santos, T. G., Rayane, M., & Henriques, A. T. (2017). Quantification of atropine in leaves of atropa belladonna: Development and validation of method by high-perfomance liquid chromatography (HPLC). Drug Analytical Research, 1, 44-49. https://doi.org/10.22456/2527-2616.74150
- Lerman, C., Schnoll, R. A., Hawk, L. W., Cinciripini, P., George, T. P., Wileyto, E. P., Swan, G. E., Benowitz, N. l, Heitjan, D. F., & Tyndale, R. F. (2015). Use of the nicotine metabolite ratio as a genetically informed biomarker of response to nicotine patch or varenicline for smoking cessation: A randomised, double-blind placebo-controlled trial. The Lancet Respiratory Medicine, 3, 131–138. https://doi.org/10.1016/S2213-2600(14)70294-2
- Lewis, R. S. (2019). Potential Mandated Lowering of Nicotine Levels in Cigarettes: A Plant Perspective. Nicotine & Tobacco Research, 21, 991–995. https://doi.org/10.1093/ntr/nty022
- Lewis, R. S., Bowen, S. W., Keogh, M. R., & Dewey, R. E. (2010). Three nicotine demethylase genes mediate nornicotine biosynthesis in Nicotiana tabacum L.: Functional characterization of the CYP82E10 gene. Phytochemistry, 71, 1988–1998. https://doi.org/10.1016/j.phytochem.2010.09.011
- Li, X., Liu, F., Wang, H., He, F., Yang, R., & Zhao, M. (2019). Gas Chromatography-Mass Spectrometry Method for Simultaneous Detection of Nine Alkaloids in Tobacco and Tobacco Products by QuEChERS Sample Preparation. Analytical Sciences: The International Journal of the Japan Society for Analytical Chemistry, 35, 849–854. https://doi.org/10.2116/analsci.19P063
- Liu, Y., & Liu, C. (2015). Determination of Total Alkaloids in Different Parts of Actinidia arguta by Spectrophotography. International Conference on Materials, Environmental and Biological Engineering (MEBE 2015) https://doi.org/10.2991/mebe-15.2015.31
- Murray, J. (2014). Nicotine and what Else?: HPLC Elution Optimization for the Analysis of Alkaloids Found in Electronic Cigarettes. University of Tennessee at Chattanooga, United States.
- Perfetti, T. A., Dube, M. F., & Coleman, W. M. (2022). Qualitative and Quantitative Analysis of Nicotine, Nicotine Derivatives, and Nicotine-Related Alkaloid Optical Isomers: A Review. Contributions to Tobacco & Nicotine Research, 31, 175–185. https://doi.org/10.2478/cttr-2022-0018
- Petruczynik, A. (2012). Analysis of alkaloids from different chemical groups by different liquid chromatography methods. Open Chemistry, 10, 802–835. https://doi.org/10.2478/s11532-012-0037-y
- Popova, V., Ivanova, T., Stoyanova, A., Georgiev, V., Hristeva, T., Nikolova, V., Docheva, M., Nikolov, N., & Damyanova, S. (2018). Phytochemicals in leaves and extracts of the variety “Plovdiv 7” of Bulgarian oriental tobacco (Nicotiana tabacum L.). Trends in Phytochemical Research, 2, 27–36.
- Saitoh, F., Noma, M., & Kawashima, N. (1985). The alkaloid contents of sixty Nicotiana species. Phytochemistry, 24, 477–480. https://doi.org/10.1016/S0031-9422(00)80751-7
- Salamah, N., & Ningsih, D. S. (2017). Total alkaloid content in various fractions of Tabernaemonata sphaerocarpa Bl. (Jembirit) leaves. IOP Conference Series: Materials Science and Engineering. https://doi.org/10.1088/1757-899X/259/1/012017
- Shikanga, E. A., Kamatou, G. P. P., Chen, W., Combrinck, S., & Viljoen, A. M. (2012). Validated RP-UHPLC PDA and GC–MS methods for the analysis of psychoactive alkaloids in Sceletium tortuosum. South African Journal of Botany, 82, 99–107. https://doi.org/10.1016/j.sajb.2012.05.004
- Shoji, T., Kajikawa, M., & Hashimoto, T. (2010). Clustered Transcription Factor Genes Regulate Nicotine Biosynthesis in Tobacco. The Plant Cell, 22, 3390–3409. https://doi.org/10.1105/tpc.110.078543
- Singh, J. (2015). International conference on harmonization of technical requirements for registration of pharmaceuticals for human use. Journal of Pharmacology & Pharmacotherapeutics, 6, 185–187. https://doi.org/10.4103/0976-500X.162004
- Sun, B., Tian, Y.-X., Zhang, F., Chen, Q., Zhang, Y., Luo, Y., Wang, X.-R., Lin, F.-C., Yang, J., & Tang, H.-R. (2018). Variations of Alkaloid Accumulation and Gene Transcription in Nicotiana tabacum. Biomolecules, 8, 114. https://doi.org/10.3390/biom8040114
- Tassew, Z., & Chandravanshi, B. S. (2015). Levels of nicotine in Ethiopian tobacco leaves. SpringerPlus, 4, 649. https://doi.org/10.1186/s40064-015-1448-y
- Tayoub, G., Sulaiman, H., & Alorfi, M. (2016). Determination of nicotine levels in the leaves of some Nicotiana tabacum varieties cultivated in Syria. Herba Polonica, 61, 23–30. https://doi.org/10.1515/hepo-2015-0028
- Tengs, T. O., Ahmad, S., Savage, J. M., Moore, R., & Gage, E. (2005). The AMA proposal to mandate nicotine reduction in cigarettes: A simulation of the population health impacts. Preventive Medicine, 40, 170–180. https://doi.org/10.1016/j.ypmed.2004.05.017
- Tepecik, M., & Ongun, A.R. (2020). Determination of some quality parameters of oriental type tobacco based on harvest times. Turkish Journal of Agricultural Research, 7(2), 156-162. https://doi.org/10.19159/tutad.691727
- Tiwari, R. K., Sharma, V., Pandey, R. K., & Shukla, S. S. (2020). Nicotine Addiction: Neurobiology and Mechanism. 23, 1–7. https://doi.org/10.3831/KPI.2020.23.001
- Wang, S. S., Shi, Q. M., Li, W. Q., Niu, J. F., Li, C. J., & Zhang, F. S. (2008). Nicotine concentration in leaves of flue-cured tobacco plants as affected by removal of the shoot apex and lateral buds. Journal of Integrative Plant Biology, 50, 958-964. https://doi.org/10.1111/j.1744-7909.2008.00684.x
- Yang, S. S., Smetena, I., & Huang, C. B. (2002). Determination of tobacco alkaloids by gas chromatography with nitrogen-phosphorus detection. Analytical and Bioanalytical Chemistry, 373, 839–843. https://doi.org/10.1007/s00216-002-1380-1
- Yasinok, A. E., Sahin, F. I., Eyidogan, F., Kuru, M., & Haberal, M. (2009). Grafting tomato plant on tobacco plant and its effect on tomato plant yield and nicotine content. Journal of the Science of Food and Agriculture, 89, 1122–1128. https://doi.org/10.1002/jsfa.3555
- Zahreddine, Z., Jaber, A., Haidar, S. A., Hosri, C., Ibrahim, G., & Cheble, E. (2021). HPLC-DAD multi-residue method for determination of florfenicol, penicillin and tetracycline residues in raw cow milk. Journal of Clinical and Laboratory Research, 2, 1–8.
- Zhang, J., Ji, H., Sun, S., Mao, D., Liu, H., & Guo, Y. (2007). Selective Determination of Pyridine Alkaloids in Tobacco by PFTBA Ions/Analyte Molecule Reaction Ionization Ion Trap Mass Spectrometry. Journal of the American Society for Mass Spectrometry, 18, 1774–1782. https://doi.org/10.1016/j.jasms.2007.07.017