根據流行病理學調查顯示造成人類致癌的因素中飲食占了60%。其中一種被US-EPA列為B2致癌物亞硝胺(N-nitrosamines)是令人關注探討的一群化合物，亞硝胺這一類化學物質具有高毒性、誘發突變及致癌的性質。近30年來，許多研究指出食物中含有亞硝化合物，主要因為亞硝胺容易由亞硝酸鹽或硝酸鹽與二級胺類發生胺化反應而產生，例如本研究的醃製肉品等加工食品。
本研究主要是發展出一套快速、溶劑量少且高通量的樣品前處理方法。使用微波輔助萃取(microwave-assisted extraction，MAE)及搭配分散微固相萃取法(dispersive micro-solid phase extraction，D-µ-SPE)，一次可萃取30組肉類樣品中的亞硝胺。本研究除了開發新的檢測肉品中亞硝胺的技術外，並使用實驗統計設計(statistical experimental design)及變異數分析(analysis of variance，ANOVA)來做最佳化的探討。將肉品中亞硝胺萃取到水溶液裡，其中因子為時間及溫度，最佳化結果為10分鐘、100 C。而在D-µ-SPE方面，選用100 mg 商業化分子篩-Carboxen 1000，搖盪30分鐘完成萃取後，快速過濾吸附劑以200 μL的二氯甲烷回溶。以10 μL大體積進樣方式導入氣相層析儀，可測得亞硝胺方法偵測極限至0.006 ng/g。檢測市面上肉類產品其萃取相對標準偏差小於20%。

The presence of N-nitrosamines (NAms) in food stuff has become a significant issue on Public health in last decades, NAms are throught to be mutagenic and carcinogenic with action on the liver, kidney and lung. These compounds are formed by reaction of N-containing substances. In this study, a sensitive procedure, microwave-assisted extraction (MAE) coupled dispersive micro solid-phase extraction (D-μ-SPE), was developed to extract N-nitroso-
dimethylamine (NDMA) and other six volatile N-nitrosamines (NAms) from meat products. The parameters affecting the MAE and D-μ-SPE efficiency were systematically investigated. For MAE, 5-g of pasted meat sample was extracted in 30 mL sodium hydroxide (0.025 M) solution at 100 C for 10 min. The best D-μ-SPE conditions were immersing 100 mg of Carboxen 1000 adsorbent in MAE extract, after 30 min of extraction by vigorously shaking, the NAms were then desorbed by 200 μL of dichloromethane. A 10 μL aliquot was determined by gas chromatography with chemical ionization mass spectrometry (GC-CI-MS) using the selected-ion -storage (SIS) mode. The limits of detection (LODs) were 0.006−0.12 ng/g.

1. 王乃龍，科學月刊，1980，11月，131期。
2. 朱建華、徐揚、王英、周仕祿、周春芳，物理化學報，2004年，第二十期，946-952。
3. 陳財富，以吸附劑作為透水性反應牆材料處理地下水中MTBE 之研究，2001。
4. 曾新華、丁望賢，液相化學游離法之原理與在分析化學上的應用，科儀新知，1998。
5. 曾新華、丁望賢，離子阱質譜儀，儀器總覽，1998。
6. 程立強、劉應亮、張靜嫺、袁定勝、徐常威、孫廣輝，化學進展，2006，第十期，18卷，1298-1304。
7. 陳家揚，液相層析/質譜儀的基質效應與因應之道，2007，第二期，第六十五卷，151-155。
8. Andersen, H. J.;Skibsted, L. H., Kinetics and mechanism of thermal oxidation and photooxidation of nitrosylmyoglobin in aqueous solution. J. Agric. Food. Chem., 1992, 40,1741–1750.
9. Ansorena, D.; Montel, M. C.; Rokka, M.; Talon, R.; Eerola, S.; Rizzo, A., Analysis of biogenic amines in northern and southern European sausages and role of flora in amine production. Meat Sci., 2002, 61, 14, 1–147.
10. Anderson, L. M et al., Transplacental initiation of liver, lung, neurogenic, and connective tissue tumors by N-nitroso compounds in mice. Fundam. Appl. Toxicol., 1989, 12, 604-620.
11. Andrade, R.; Reyes, F.G.R.; Rath, S., A method for the determination of volatile N–nitrosamines in food by HS–SPME–GC–TEA. Food Chem., 2005, 91, 173–179.
12. Anastassiades, M.; Lehotay, S. J.; Stajnbaher, D.; Schenck, F. J., Fast and easy multiresidue method employing acetonitrile extrac-tion/partitioning and "dispersive solid phase extraction" for thedetermination of pesticide residues in produce. J. AOAC. Int., 2003, 86(2), 412-431.
13. Arinc, E.; Arslan, S.; Bozcaarmutlu, A.; Adali, O., Effects of diabetes on rabbit kidney and lung CYP2E1 and CYP2B4 expression and drug metabolism and potentiation of carcinogenic activity of N-nitrosodimethylamine in kidney and lung. Food Chem. Toxicol., 2007, 45, 107-118.
14. Belitz, H. D.; Grosch W., Lehrbuch des Lebensmittelchemie.(4th Ed)., Berlin:Springer ,1992.
15. Cardenes, L.; Ayala, J.H.; Gonzalez, V.; Afonso, A.M. Fast microwave–assisted dansylation of N-nitrosamines – Analysis by high-performance liquid chromatography with fluorescence detection. J. Chromatogr. A, 2002, 946, 133–140.
16. Casa-Lillo, M.A.; Alcaniz-Monge, J.; Raynumdo-Pinero, E.; Cazorla-Amoros, D.; Linares-Solano, A., Molecular sieve properties of general-purpose carbon fibres. Carbon, 1998, 36, 1353-1360.
17. Careri, M.; Corradini, C.; Elviri, L.; Mangia, A., Optimization of a rapid microwave assisted extraction method for the liquid chromatography–electrospray-tandem mass spectrometry determination of isoflavonoid aglycones in soybeans., J. Chromatogr. A, 2007, 1152, 274–279.
18. Denton, J. E. et al., Public Health Goal for N-Nitrosodimethylaamine in Drinking Water, California Public Health Goal, 2006.
19. Diaz Gomez, M. I.; Swann, P.F.; Magee, P.N., The absorption and metabolism in rats of small oral doses of dimethylnitrosamine. Biochem. J., 1977, 164, 497-500.
20. Doll, R.; Peto, R., The causes of cancer:quantitative estimates of avoidable risk of cancer in the United States today. J. Natl. Cacncer Inst. 1981, 66, 1191-130.
21. Donike, M.; Geyer, H.; Gotzmann, A.; Mareck-Engelke, U., Sport und Buch Strauβ, 1996, 277-284.
22. Drabik-Markiewicz, G.; Dejaegher, B.; De Mey, E.;Kowalsk, T.;Paelinck, H.; Vander Heyden, Y., Influence of putrescine, cadaverine, spermidine or spermine on the formation of N-nitrosamine in heated cured pork meat, Food Chem., 2011, 126, 1539–1545.
23. Drabik-Markiewicz, G.; Van den Maagdenberg , K.; De Mey, E.; Deprez, S.; Kowalska, T.; Paelinck, H., Role of proline and hydroxyproline in N-nitrosamine formation during heating in cured meat. Meat Sci., 2009, 81, 479–486.
24. Eisenbrand, G.;Jancowski, C.;Preussmann, R., Analysis, formation and occurrence of volatile and non-volatile N-nitrosocompounds: Recent results. In Proceedings II International Symposium on Nitrite in Meat Products, 1977, 155–169.
25. EPA., US Environmental Protection Agency, NNitrosodimethylamine,
甲、 CASRN 62–75–9. IRIS, 1997. Available at http://www.epa.gov/iris/ subst/0045.htm.
26. EPA Method 521., Determination of nitrosamines in drinking water by solid phase extraction and capillary column gas chromatography with large volume injection and chemical ionization tandem mass spectrometry (MS/MS).2004.
27. Fazio, T.; White, R. H.; Howard, J. W., Analysis of nitrite-and/or nitrate-processed meats for N-nitrosodimethylamine. J. Assoc. Off. Anal. Chem, 1971, 54, 1157-1159.
28. Filhoa, P. J. S.; Riosb, A.; Miguel, V.; Kelen, D. Z., Determination of nitrosamines in preserved sausages by solid-phase extraction–micellar electrokinetic chromatography. J. Chromatogr. A, 2003, 985, 503–512.
29. Forman, D., Dietary exposure to N-nitroso compounds and the risk of human cancer. Cancer Surveill.,1987, 6, 719–738.
30. Fu, S.C.; Tzing, S.H.; Chen, H.C.; Wang, Y.; Ding, W.H., Dispersive micro-solid phase extraction combined with gas chromatography–chemical ionization mass spectrometry for the determination of N-nitrosamines in swimming pool water samples. Anal Bioanal Chem , 2012, 402, 2209–2216.
31. Gasarasi, G.; Troy, P.D.; Debeuckelaere, W.; Delvaux, F.; Derdelinckx, G., Les N-nitrosamines volatiles dans l’alimentation: La viande. Journal de Pharmacie de Belgique, 2001, 56, 51–53.
32. Greenfield, E.L.; Smith, W.J.; Malanoski, A.J., Mineral–oil vacuum distillation method for nitrosamines in fried bacon,with thermal-energy analyzer– Collaborative study. J. Assoc. Off. Anal. Chem., 1982, 65, 1319–1332.
33. Haldane, J., The red color of salted meat. J. Hygiene, 1901, I, 115–122.
34. Honikel, K.,The use and control of nitrate and nitrite for the processingof meat products. Meat Sci., 2008, 78, 68.
35. Hubschmann, H. J., Handbook of GC/MS: Fundamentals and Applications, 2th ed, Wiley-VCH, 2008.
36. Iris Substance List, U.S. Environmental Protection Agency, 2010,
甲、 http://www.epa.gov/iris/subst/index.html.
37. Jurado–Sanchez B.; Ballesteros E.; Gallego M., Gas chromatographic determination of N-nitrosamines in beverages following automatic solid–phase extraction. J. Agric. Food. Chem., 2007, 55, 9758–9763.
38. Jurado, B.; Ballesteros, E.; Gallego, M., Comparison of several solid-phase extraction sorbents for continuous determination of amines in water by gas chromatography-mass spectrometry. Talanta, 2009,79, 613-620,
39. Kalač, P., Biologically active polyamines in beef, pork and meat products: A review. Meat Sci., 2006, 73, 1–11.
40. Knappe, D., Effects Of Activated Carbon Characteristics On Organic Contaminant Removal, AwwaRF report number 99926F, 2003.
41. Li, L., Effects of activated carbon surface chemistry and pore structure on the adsorption of trace organic contaminants from aqueous solution., 2002.
42. Lilov, S. K.,Determination of the Effective Kinetic Diameter of the Complex Molecules. Cryst. Res. Technol., 1986, 10, 1299-1302.
43. Lijinsky, W.; Epstein, S., Nitrosamines as environmental carcinogens. Nature, 1970, 225, 21.
44. Liu, R. H.; Conboy, J. J.; Hotchkiss, J. H., Nitrosation by nitro- nitroso derivatives of oleifins: a potential mechanism for Nnitrosamine formation in fried bacon. J. Agric. Food. Chem., 1988, 36 , 984.
45. Liviac, D.; Creus, A.; Marcos, R.,Genotoxic evaluation of the non-halogenated disinfection by-products nitrosodimethylamine and nitrosodiethylamine. J. Hazard. Mater, 2011, 185, 613-618.
46. Loeppky, R.N.; Hansen, J.J. ; Keefer, L.K. Reducing nitrosamine contamination in cutting fluids. Food chem. Toxicol, 1983, 5, 607–613
47. Magee, P. N.; Barnes, J. M., The production of malignant primary hepatic tumours in the rat by feeding dimethylnitrosamine. Bri. J. Cancer, 1956, 10, 114–122.
48. Marion, D.; Gladys, B.; Janice, M; Patricia, B.; Stephen, H.; Preston-Martin, S., A review:dietary and endogenously formed N-nitroso compounds and risk of chikdhood brain tumors. Cancer Causes and Control, 2005, 16, 619-63.
49. Mavelle, T.; Bouchikhi B.; Debry G. The occurrence of volatile
甲、 N-nitrosamines in French foodstuffs. Food Chem., 1991, 42, 321–338.
50. Mirvish, S. S., Blocking formation of N-nitroso compounds with ascorbic acid in vivo and in vitro. Ann. N. Y. Acad. Sci., 1975, 258, 175–180.
51. Mirvish, S. S., Role of N-nitroso compounds (NOC) and N-nitrosation in etiology of gastric, esophageal and bladder cancer and contribution to cancer of known exposures to NOC. Cancer Lett., 1995 , 93, 17-48.
52. Nawrocki, J.; Andrzejewski, P., Nitrosamines and water. J. Hazard. Mater., 2011, 189, 1-18.
53. Özel, M.Z.; Gogus, F.; Yagci, S.; Hamilton, J.F.; Lewis, A.C., Determination of volatile nitrosamines in various meat products using comprehensive gas chromatography – nitrogen chemiluminescence detection. Food Chem. Toxicol., 2010, 48, 3268-3273.
54. Pensabene, J.W.; Fiddler, W., Dual column chromatographic method for determination of N-nitrosothiazolidine in fried bacon. J. Assoc. Off. Anal. Chem., 1982, 65, 1346–1349.
55. Preussmann, R; Stewart, B.W., Chemical Carcinogens. American Chemical Society, 1984, 643-828.
56. Reinik, M.; Tamme, T.; Roasto, M.;Juhkan, K.; Jurtśenko, S.;Tenńo,T.;Kiis, A., Nitrites, nitrates and N-nitrosoamines in Estonian cured meat products:Intake by Estonian children and adolescents. Food Addit. Contam., 2005; 22(11), 1098–1105.
57. Ruiz-Capillas, C.; Jimenez-Colmenero, F., Biogenic amines in meat and meat products. Crit. Rev. Food Sci. Nutrition., 2004, 44, 489–499.
58. Rywotycki, R., Meat nitrosamine contamination level depending on animal breeding factors. Meat Sci., 2003, 65, 669–676.
59. Rywotycki, R., Nitrosamine concentrations in beef ham:Influence of smoking and diversified combinations of functional additives. Fleischwirtsch. Int., 2001, 2(5), 77–80.
60. Sen, N. P., Recent studies in Canada on the occurrence and formation of N-nitroso compounds in foods and food contact materials. IARC Sci. Publications., 1991, 105, 232-234.
61. Sen, N. P., Seaman S.W., Page B.D., Rapid semi–quantitative estimation of N-nitrosodibutylamine and N-nitrosodibenzylamine in smoked hams by solid-phase microextraction followed by gas chromatography-thermal energy analysis. J. Chromatogr. A, 1997, 788, 131–140.
62. Sergei, J.; Toomas, T.; Uldo, M.; Mari, R., Determination of volatile N-nitrosamines by gas chromatography–mass spectrometry with positive-ion chemical ionization. Proc. Estonian Acad. Sci. Chem, 2002, 51, 3, 169–184.
63. Sigma-Aldrich (http://www.sigmaaldrich.com/taiwan.html)
64. Skoog, D. A.; Holler, F. J.; Nieman, T. A., Principles of instrumental analysis. 5th ed., Thomoson Learning, 2002.
65. Skrypec, D. J.; Gray, J. I.; Mandagere, A. K.; Booren, A. M.;Pearson,M.; Cuppet, S.L., Effects of bacon composition and processing of N-nitrosamine formation. Food Technol., 1985, 39, 74–79.
66. Sokolova, I. A.; Lusternik, B. E., Numerical analysis of density dependence for effective transport kinetic diameter of real fluids. Lecture Notes in Computer Sci., 1997, 1196, 474-481.
67. Thepkaew, J.; Therdthianwong, A.; Therdthianwong, S., Key parameters of active layers affecting proton exchange membrane (PEM) fuel cell performance . Energy, 2008, 33, 1794–1800.
68. Stuff Janice, E.; Goh Eugenia, T.; Barrera Stephanie, L.; Bondy Melissa, L.; Forman Michele, R., Construction of an N-nitroso database for assessing dietary intake. J. Food Compos. Anal., 2009, 22 ,42–47.
69. The determination of N- nitrosamines infood. Quality Assurance and safety of Crops & Foods, 2010, ISSN 1757-8361.
70. Tricker, A.R.; Preussmann, R.; Carcinogenic N-nitrosamines in the diet:occurrence, formation, mechanisms and carcinogenic potential. Mutat.Res.,1991, 259, 277–289.
71. US-EPA, IRIS substance File, 1987( http://www.epa.gov/IRIS/).
72. US-FDA.,Chapter III –Food Safety and Inspection Service, Department of
i. Agriculture. 2005. Available at http://cfr.vlex.com/vid/22-certain-other- permitted-uses-19611025.
73. US-FDA Code of Federal Regulations – Title 21: Food and Drugs – 21 CFR 179.26 – Ionizing radiation for the treatment of food. 2005 . Available at http://cfr.vlex.com/vid/22- certain-other-permitted-uses-19611025.
74. Wei, F.; Xua, X.; Zhou, G.; Zhao, G.; Li, C.; Zhang, Y.; Lingzhen, C.; Jun, Q., Irradiated Chinese Rugao ham: Changes in volatile N-nitrosamine, biogenic amine and residual nitrite during ripening and post-ripening., Meat Sci., 2009, 81, 451–455.
75. Wexler, P.; Anderson, B.; Peyster, A.; Gad, S.; Hakkinen, P. J.; Kamrin, M.; Locey, B.; Mehendale, H.; Pope, C.; Shugart, L., Encyclopedia of Toxicol., 2005, Second Edition, 255-258.
76. Wyllie, T.; Morehouse, L., Mycotoxic fungi, mycotoxins, mycotoxicoses. N. Y. Basel: Marcel Dekker Inc., 1990–1991.
77. Yurchenko, S.; Molder, U.; The occurrence of volatile N-nitrosamines in Estonian meat products. Food Chem., 2007, 100, 1713–1721.
78. Yurchenko, S., Molder, U.,N-nitrosodimethylamine analysis in Estonian beer using positive-ion chemical ionization with gas chromatography mass spectrometry. Food Chem., 2005, 89, 455–463.
79. Zhao, Y.; Boyd, J.; Woodbeck, M.; Andrews, R.; Hrudey, S.E.; Li, X.-F., Formation of N-nitrosamines from eleven disinfection treatments of seven different surface waters. Environ. Sci. Technol., 2008, 42, 4857-4862.
80. Zhao, Y.; Wang, J; Luan, Z.; Peng, X.; Liang, Z.; Shi, L., Removal of phosphate from aqueous solution by red mud using a factorial design. J. Hazardous Mater., 2009, 165, 1193–119.