毛细管区带电电泳分离测定烟草生物碱
摘要
生物碱是烟草的重要成分。其中，烟碱含量最高，其它生物碱含量很低。有些生物碱能影响卷烟的品质。另外，卷烟在燃吸时，会产生烟草特有的亚硝胺，这些亚硝胺化合物是吸烟者易患肺、食管和口腔癌的原因。因此，浓缩和检测烟草样品中的生物碱(包括去甲基烟碱、烟碱、米渥斯明和假木贼碱等)是必要的。
近年来，已有一些关于烟草生物碱检测方法的报道，它们包括气相色谱，液相色谱和毛细管电泳法等。
毛细管电泳具有简单、快速、经济和环境友好的特点，遗憾的是用磷酸缓冲体系的毛细管区带电泳未能将米渥斯明与其它生物碱分离。本文采用酒石酸缓冲体系，成功地分离了包括米渥斯明在内的四种烟草生物碱。该缓冲体系不仅提高了分离度，还改善了检测灵敏度。为了检测烟草中微量生物碱，论文研究并采用了阳离子耗尽富集法进样。
在水溶液中，烟草生物碱存在三种质子化形态，为了使生物碱主要以双质子化形态存在，论文采用pH 2.8～3.2，360～410 mmol／L的酒石酸缓冲体系。与磷酸缓冲体系相比，这种高浓度低pH值的酒石酸缓冲体系能更有效地分离和检测烟草生物碱。
在对卷烟中生物碱进行定量测定时，仔细调查了卷烟生物碱的提取条件以及缓冲溶液的最佳pH值和浓度。测定方法的检出限优于已有的烟草生物碱毛细管电泳分析法。
Alkaloid is one of the most important components in tobacco. Among tobacco alkaloids, the concentration of nicotine is the highest and those of other alkaloids are quite low. Some alkaloids influence the quality of cigarette. In addition, the tobacco alkaloids can generate specific tobacco nitrosamines during smoking, which may be the causative factor of lung, esophagus and oral cavity cancers in the smoking population. Therefore, it is necessary to concentrate and determine tobacco alkaloids in cigarette samples, including nicotine, nornicotine, myosmine and anabasine etc.
In recent years, some determination methods of tobacco alkaloids were reported, including gas chromatography, high-performance liquid chromatography and capillary electrophoresis etc.
Capillary electrophoresis is a simple, rapid, low-cost and environment-friendly separation technique. However, only a few paper reported the separation of tobacco alkaloids except myosmine with capillary electrophoresis. In this dissertation, the separation of tobacco alkaloids was investigated and an effective separation method with capillary zone electrophoresis (CZE) was proposed, which was capable of separating more alkaloids including myosmine in tobacco samples. It was found that the tartaric buffer solution at low pH provided higher separation efficiency and detection sensitivity comparing with those of phosphate buffer solution. In addition, cation-selective exhaustive injection, as an on-column preconcentration method of CZE, was also investigated for the analysis of trace alkaloids in cigarette samples.
Tobacco alkaloids have three protonated species depending on the pH value of
近年来，已有一些关于烟草生物碱检测方法的报道，它们包括气相色谱，液相色谱和毛细管电泳法等。
毛细管电泳具有简单、快速、经济和环境友好的特点，遗憾的是用磷酸缓冲体系的毛细管区带电泳未能将米渥斯明与其它生物碱分离。本文采用酒石酸缓冲体系，成功地分离了包括米渥斯明在内的四种烟草生物碱。该缓冲体系不仅提高了分离度，还改善了检测灵敏度。为了检测烟草中微量生物碱，论文研究并采用了阳离子耗尽富集法进样。
在水溶液中，烟草生物碱存在三种质子化形态，为了使生物碱主要以双质子化形态存在，论文采用pH 2.8～3.2，360～410 mmol／L的酒石酸缓冲体系。与磷酸缓冲体系相比，这种高浓度低pH值的酒石酸缓冲体系能更有效地分离和检测烟草生物碱。
在对卷烟中生物碱进行定量测定时，仔细调查了卷烟生物碱的提取条件以及缓冲溶液的最佳pH值和浓度。测定方法的检出限优于已有的烟草生物碱毛细管电泳分析法。
Alkaloid is one of the most important components in tobacco. Among tobacco alkaloids, the concentration of nicotine is the highest and those of other alkaloids are quite low. Some alkaloids influence the quality of cigarette. In addition, the tobacco alkaloids can generate specific tobacco nitrosamines during smoking, which may be the causative factor of lung, esophagus and oral cavity cancers in the smoking population. Therefore, it is necessary to concentrate and determine tobacco alkaloids in cigarette samples, including nicotine, nornicotine, myosmine and anabasine etc.
In recent years, some determination methods of tobacco alkaloids were reported, including gas chromatography, high-performance liquid chromatography and capillary electrophoresis etc.
Capillary electrophoresis is a simple, rapid, low-cost and environment-friendly separation technique. However, only a few paper reported the separation of tobacco alkaloids except myosmine with capillary electrophoresis. In this dissertation, the separation of tobacco alkaloids was investigated and an effective separation method with capillary zone electrophoresis (CZE) was proposed, which was capable of separating more alkaloids including myosmine in tobacco samples. It was found that the tartaric buffer solution at low pH provided higher separation efficiency and detection sensitivity comparing with those of phosphate buffer solution. In addition, cation-selective exhaustive injection, as an on-column preconcentration method of CZE, was also investigated for the analysis of trace alkaloids in cigarette samples.
Tobacco alkaloids have three protonated species depending on the pH value of
引文
[1] 左天觉著，朱尊权等译，烟草的生产，生理和生物化学，上海远东出版社，1993，pp．308，310
[2] De Bardeleben M Z, Dictionary of Tobacco Terminology, Second Edition, Philip Morris, New York, 1987, p. 4
[3] Garner W W, The Production of Tobacco, Revised First Edition, Blakiston Co., Philadelphia, 1951, pp. 313,433,435
[4] Lewis L J, Lamb J D, Eatough D J, Hanson L D, Lewis E A, J. Chromatogar. Sci., 1990, 28:200-203
[5] Enzell C R, Wahlberg I, Recent Advances in Tobacco Science, 1980, 6:64-122
[6] Secor H V, Edwards W B, J. Org. Chem., 1979, 44 (18): 3136-3140
[7] James W O, Alkaloids in plants, Endeavor, 1953, 12:76-79
[8] Hecht S S, Chen C H B, Ornaf R M, Jacobs E, Adams J D, Hoffmann D, J. Org. Chem., 1978, 43 (1): 72-76
[9] Djordjevic M V, Gay S L, Bush L P, Chaplin J F, J. Agr. Food Chem., 1989, 37 (3): 752-761
[10] 辛惠君，谢复炜，邓大君，分析化学，2000，48(4)：436-438
[11] Hecht S S, Mutation Research, 1999, 424:127-142
[12] Kleinsasser N H, Wallner B C, Harreus U A, Zwickenpflug W, Richter E, Toxicology, 2003, 192:171-177
[13] Gottscho A M, Lin J L,Duck W N, Losty T A, J. Assoc. Off. Anal. Chem., 1988, 71:1110-1115
[14] Raisi A, Alipour E, Manouchehri S, Chromatographia, 1986, 21:711-712
[15] Popl M, Fahnrich J, Vlastimil T, Chromatographic Analysis of Alkaloids, Marcel Dekker Inc., New York, 1990, pp. 313-315
[16] Saunders J A, Blume D E, J. Chromatogr., 1981,205:147-154
[17] Manceau F, Fliniaux M A, Jacquin-Dubreuil A, Phytochem.Anal., 1992, 3:65-68
[18] Yang S S, Smetena I, Chromatographia, 1995, 40:375-378
[19] International Standards Organization, International Standard ISO 2881, Geneva, Switzerland, 1992
[20] Official Methods of Analysis, Method 960.08, 16th Ed. AOAC INTERNATIONAL, Gaithersburg, MD, 1997
[21] Burns D T, Collin E J, J. Chromatogr., 1977, 133:378-381
[22] Severson R F, McDuffie K L, Arrendale R F, Gwynn G R, Chaplin J F, Jonhson A W, J. Chromatogr., 1981, 211:111-121
[23] Jassie L., Revesz R., Kierstead T., Hasty E., Matz S., Microwave-Enhanced Chemistry: Fundamentals, Sample Preparation and Applications, Eds Kingston H. M., Haswell S. J., American Chemical Society, Washington DC, 1997, pp 569-609.
[24] Ganzler K, Szinai I, Salg6 A, J. Chromatogr., 1990, 520:257-262
[25] Pare J. R. J., Sigouin M., Lapointe J., 1991, U. S. Patent No.5002784
[26] Ganzler K, Salgd A, Valk6 K, J. Chromatogr., 1986, 371:299-306
[27] Loperz-Avila V, Young R, Teplitsky N, J. AOAC Int., 1996, 79:142-156
[28] Pare J R J, Matini G, Bélanger J M R, Li K, Rule C, Thibert B, Yaylayan V, Liu Z, Mathé D, Jacquault P, J. AOAC Int., 1997, 80:928-933
[29] Bichi C, Beliarab F F, Rubiolo P, 1992, Lab 2000, 6:36-38
[30] Cundiff R H, Markunas P C, J. AOAC Int., 1960, 43:519-524
[31] Lourenco M.G. (1985) Ph.D Thesis, Instituto Superior de Agronomia. Lisbon, Portugal
[32] Etal J, Jones N M, Bernardo-Gil M G, Lourenco M G, J. AOAC Int., 2001, 84 (2): 309-316
[33] 谢振伟，但德忠，赵燕，周娜，化学通报，2005，68(w090)：1-11
[34] 何梅，夏之宁，郭宝元，王力，化学通报，2002，65(w018)：1-4
[35] Ogg C L, Bates W W, Cogbill E, Harrow L S, Petersen E L, J. AOAC, 1960, 32: 524
[36] Piade J J, Hoffmann D, J. Liq. Chromatogr., 1980, 3 (10): 1505
[37] Hariharan M, VanNoord T, Greden J F, Clin. Chem., 1988, 34 (4): 724
[38] Cundy K C, Crook PA, J. Chromatogr., 1984, 306:291
[39] Cai J B, Liu B Z, Lin P, Su Q D, J. Chromatogr. A, 2003, 1017:187-193
[40] Sudan B J L, Brouillard C, Strehler C, Strub H, Sterboul J, Sainte-Laudy J, J. Chromatogr., 1984, 288:415
[41] Armstrong D W, Bertrand G L, Ward K D, Secor H V, Seeman J I, Anal. Chem., 1990, 62:332
[42] 单卫青，李华儒，分析测试通报，1992，11(5)：53-55
[43] Lu G H, Ralapati S, Electrophoresis, 1998, 19:19-26
[44] Yang S S, Smetena I, Goldsmith A I, J Chromatogr. A, 1996, 746: 131-136
[1] Hjerten S, Chromatgr. Rev., 1967, 9:122
[2] J.W. Jorgenson, K.D. Lukacs, Anal. Chem. 1981, 53:1298
[3] Terabe S, Otsuka K, Ichikawa K, Anal.Chem., 1984, 56:111
[4] Hjerten S, Liao J, Yao K J, Chromatgr., 1987, 387:127
[5] Cohen A, Karger B L J, Chromatogr., 1987, 397:409
[6] Rose D J, Jorgenson J W, Anal.Chem., 1988, 60:642
[7] Jones W R, Jandik P, American Laboratory, 1990, 22:51
[8] Proceedings of the Sixth Annual Frederick Conference on Capillary Electrophoresis. J. Chromatogr. B, 1996 683: 1-131.
[9] Ralapati S, J. Chromatogr. B, 1997,685:117-129.
[1] Yang S S, Smetena I, Chromatographia, 1995, 40:375-378
[2] Lu G H, Ralapati S, Electrophoresis, 1998, 19:19-26
[3] Joselito P. Quirino, Shigeru Terabe, Anal. Chem., 2000, 72:1023-1030
[4] Chien R. L., Burgi, D. S., Anal. Chem., 1992, 64: 489A-496A
[5] Quirino, J. P., Terabe, S., Science, 1998, 282:465-468
[6] Quirino, J. P., Terabe, S., Anal. Chem., 1999, 71:1638-1644
[1] Yang S S, Smetena I, Chromatographia, 1995, 40:375-378
[2] Morie G P, Tobacco Science, 1972, 16:167
[3] Lu G H, Ralapati S, Electrophoresis, 1998, 19:19-26
[2] De Bardeleben M Z, Dictionary of Tobacco Terminology, Second Edition, Philip Morris, New York, 1987, p. 4
[3] Garner W W, The Production of Tobacco, Revised First Edition, Blakiston Co., Philadelphia, 1951, pp. 313,433,435
[4] Lewis L J, Lamb J D, Eatough D J, Hanson L D, Lewis E A, J. Chromatogar. Sci., 1990, 28:200-203
[5] Enzell C R, Wahlberg I, Recent Advances in Tobacco Science, 1980, 6:64-122
[6] Secor H V, Edwards W B, J. Org. Chem., 1979, 44 (18): 3136-3140
[7] James W O, Alkaloids in plants, Endeavor, 1953, 12:76-79
[8] Hecht S S, Chen C H B, Ornaf R M, Jacobs E, Adams J D, Hoffmann D, J. Org. Chem., 1978, 43 (1): 72-76
[9] Djordjevic M V, Gay S L, Bush L P, Chaplin J F, J. Agr. Food Chem., 1989, 37 (3): 752-761
[10] 辛惠君，谢复炜，邓大君，分析化学，2000，48(4)：436-438
[11] Hecht S S, Mutation Research, 1999, 424:127-142
[12] Kleinsasser N H, Wallner B C, Harreus U A, Zwickenpflug W, Richter E, Toxicology, 2003, 192:171-177
[13] Gottscho A M, Lin J L,Duck W N, Losty T A, J. Assoc. Off. Anal. Chem., 1988, 71:1110-1115
[14] Raisi A, Alipour E, Manouchehri S, Chromatographia, 1986, 21:711-712
[15] Popl M, Fahnrich J, Vlastimil T, Chromatographic Analysis of Alkaloids, Marcel Dekker Inc., New York, 1990, pp. 313-315
[16] Saunders J A, Blume D E, J. Chromatogr., 1981,205:147-154
[17] Manceau F, Fliniaux M A, Jacquin-Dubreuil A, Phytochem.Anal., 1992, 3:65-68
[18] Yang S S, Smetena I, Chromatographia, 1995, 40:375-378
[19] International Standards Organization, International Standard ISO 2881, Geneva, Switzerland, 1992
[20] Official Methods of Analysis, Method 960.08, 16th Ed. AOAC INTERNATIONAL, Gaithersburg, MD, 1997
[21] Burns D T, Collin E J, J. Chromatogr., 1977, 133:378-381
[22] Severson R F, McDuffie K L, Arrendale R F, Gwynn G R, Chaplin J F, Jonhson A W, J. Chromatogr., 1981, 211:111-121
[23] Jassie L., Revesz R., Kierstead T., Hasty E., Matz S., Microwave-Enhanced Chemistry: Fundamentals, Sample Preparation and Applications, Eds Kingston H. M., Haswell S. J., American Chemical Society, Washington DC, 1997, pp 569-609.
[24] Ganzler K, Szinai I, Salg6 A, J. Chromatogr., 1990, 520:257-262
[25] Pare J. R. J., Sigouin M., Lapointe J., 1991, U. S. Patent No.5002784
[26] Ganzler K, Salgd A, Valk6 K, J. Chromatogr., 1986, 371:299-306
[27] Loperz-Avila V, Young R, Teplitsky N, J. AOAC Int., 1996, 79:142-156
[28] Pare J R J, Matini G, Bélanger J M R, Li K, Rule C, Thibert B, Yaylayan V, Liu Z, Mathé D, Jacquault P, J. AOAC Int., 1997, 80:928-933
[29] Bichi C, Beliarab F F, Rubiolo P, 1992, Lab 2000, 6:36-38
[30] Cundiff R H, Markunas P C, J. AOAC Int., 1960, 43:519-524
[31] Lourenco M.G. (1985) Ph.D Thesis, Instituto Superior de Agronomia. Lisbon, Portugal
[32] Etal J, Jones N M, Bernardo-Gil M G, Lourenco M G, J. AOAC Int., 2001, 84 (2): 309-316
[33] 谢振伟，但德忠，赵燕，周娜，化学通报，2005，68(w090)：1-11
[34] 何梅，夏之宁，郭宝元，王力，化学通报，2002，65(w018)：1-4
[35] Ogg C L, Bates W W, Cogbill E, Harrow L S, Petersen E L, J. AOAC, 1960, 32: 524
[36] Piade J J, Hoffmann D, J. Liq. Chromatogr., 1980, 3 (10): 1505
[37] Hariharan M, VanNoord T, Greden J F, Clin. Chem., 1988, 34 (4): 724
[38] Cundy K C, Crook PA, J. Chromatogr., 1984, 306:291
[39] Cai J B, Liu B Z, Lin P, Su Q D, J. Chromatogr. A, 2003, 1017:187-193
[40] Sudan B J L, Brouillard C, Strehler C, Strub H, Sterboul J, Sainte-Laudy J, J. Chromatogr., 1984, 288:415
[41] Armstrong D W, Bertrand G L, Ward K D, Secor H V, Seeman J I, Anal. Chem., 1990, 62:332
[42] 单卫青，李华儒，分析测试通报，1992，11(5)：53-55
[43] Lu G H, Ralapati S, Electrophoresis, 1998, 19:19-26
[44] Yang S S, Smetena I, Goldsmith A I, J Chromatogr. A, 1996, 746: 131-136
[1] Hjerten S, Chromatgr. Rev., 1967, 9:122
[2] J.W. Jorgenson, K.D. Lukacs, Anal. Chem. 1981, 53:1298
[3] Terabe S, Otsuka K, Ichikawa K, Anal.Chem., 1984, 56:111
[4] Hjerten S, Liao J, Yao K J, Chromatgr., 1987, 387:127
[5] Cohen A, Karger B L J, Chromatogr., 1987, 397:409
[6] Rose D J, Jorgenson J W, Anal.Chem., 1988, 60:642
[7] Jones W R, Jandik P, American Laboratory, 1990, 22:51
[8] Proceedings of the Sixth Annual Frederick Conference on Capillary Electrophoresis. J. Chromatogr. B, 1996 683: 1-131.
[9] Ralapati S, J. Chromatogr. B, 1997,685:117-129.
[1] Yang S S, Smetena I, Chromatographia, 1995, 40:375-378
[2] Lu G H, Ralapati S, Electrophoresis, 1998, 19:19-26
[3] Joselito P. Quirino, Shigeru Terabe, Anal. Chem., 2000, 72:1023-1030
[4] Chien R. L., Burgi, D. S., Anal. Chem., 1992, 64: 489A-496A
[5] Quirino, J. P., Terabe, S., Science, 1998, 282:465-468
[6] Quirino, J. P., Terabe, S., Anal. Chem., 1999, 71:1638-1644
[1] Yang S S, Smetena I, Chromatographia, 1995, 40:375-378
[2] Morie G P, Tobacco Science, 1972, 16:167
[3] Lu G H, Ralapati S, Electrophoresis, 1998, 19:19-26