Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/16867
標題: QuEChERS萃取技術結合液相層析串聯質譜術於穀類產品中真菌毒素之分析
Analysis of Mycotoxins in Grain Products by QuEChERS Combined with Liquid Chromatography-Tandem Mass Spectrometry
作者: 尤品人
Yu, Pin-Jen
關鍵字: Mycotoxin
真菌毒素
Liquid Chromatography-Mass Spectrometry
Grain product
液相層析串聯質譜術
穀類產品
出版社: 化學系所
引用: 1. M. Sulyok; F. Berthiller; R. Krska; R. Schuhmacher, Development and validation of a liquid chromatography/tandem mass spectrometric method for the determination of 39 mycotoxins in wheat and maize. Rapid Commun. Mass Spectrom. 2006, 20 (18), 2649-2659. 2. G. S. Shephard, Determination of mycotoxins in human foods. Chem. Soc. Rev. 2008, 37 (11), 2468-2477. 3. 林茂勇, 呂鋒洲, 黴菌毒素學, 民國八十年, 244-246。 4. R. Lawley; L. Curtis; J. Davis, The food safety hazard guidebook, Springer Verlag, 2008, 179-187. 5. L. S. Weckbach; E. H. Marth, Aflatoxin production by Aspergillus parasiticus in a competitive environment. Mycopathologia 1977, 62 (1), 39-45. 6. International Agency for Research on Cancer, Monograph on the Evaluation of Carcinogenic Risk to Humans, Some Naturally Occurring Substances: Food Items and Constituents, Heterocyclic Aromatic Amines and Mycotoxins. 7. B. Kabak, The fate of mycotoxins during thermal food processing. J. Sci. Food Agric. 2009, 89 (4), 549-554. 8. P. S. Steyn, Mycotoxins, general view, chemistry and structure. Toxicol. Lett. 1995, 82-83, 843-851. 9. H. S. Hussein; J. M. Brasel, Toxicity, metabolism, and impact of mycotoxins on humans and animals. Toxicology 2001, 167 (2), 101-134. 10. M. J. Sweeney; A. D. W. Dobson, Molecular biology of mycotoxin biosynthesis. FEMS Microbiol. Lett. 1999, 175 (2), 149-163. 11. C. M. Franco; C. A. Fente; B. Vazquez; A. Cepeda; L. Lallaoui; P. Prognon; G. Mahuzier, Simple and sensitive high-performance liquid chromatography-fluorescence method for the determination of citrinin application to the analysis of fungal cultures and cheese extracts. J. Chromatogr. A 1996, 723 (1), 69-75. 12. V. Gökmen; J. Acar, Simultaneous determination of 5-hydroxymethylfurfural and patulin in apple juice by reversed-phase liquid chromatography. J. Chromatogr. A 1999, 847 (1-2), 69-74. 13. B.-H. Liu; F.-Y. Yu; T.-S. Wu; S.-Y. Li; M.-C. Su; M.-C. Wang; S.-M. Shih, Evaluation of genotoxic risk and oxidative DNA damage in mammalian cells exposed to mycotoxins, patulin and citrinin. Toxicol. Appl. Pharmacol. 2003, 191 (3), 255-263. 14. A. Ciegler; R. F. Vesonder; L. K. Jackson, Produciton and biological activity of patulin and citrinin from Penicillium expansum. Appl. Environ. Microbiol. 1977, 33 (4), 1004-1006. 15. B.-j. Xu; X.-q. Jia; L.-j. Gu; C.-k. Sung, Review on the qualitative and quantitative analysis of the mycotoxin citrinin. Food Control 2006, 17 (4), 271-285. 16. V. Gökmen; J. Acar, Incidence of patulin in apple juice concentrates produced in Turkey. J. Chromatogr. A 1998, 815 (1), 99-102. 17. M. Takino; S. Daishima; T. Nakahara, Liquid chromatography/mass spectrometric determination of patulin in apple juice using atmospheric pressure photoionization. Rapid Commun. Mass Spectrom. 2003, 17 (17), 1965-1972. 18. G. Boonzaaijer; I. Bobeldijk; W. A. van Osenbruggen, Analysis of patulin in dutch food, an evaluation of a SPE based method. Food Control 2005, 16 (7), 587-591. 19. T. Shivanandappa; M. K. Krishnakumari; S. K. Majumder, Reproductive Potential of male rats fed dietary ethylene dibromide. J. Food Saf. 1987, 8 (3), 147-155. 20. M. L. Fernández-Cruz; M. L. Mansilla; J. L. Tadeo, Mycotoxins in fruits and their processed products: Analysis, occurrence and health implications. J. Adv. Res. 2010, 1 (2), 113-122. 21. G. S. Shephard; N. L. Leggott, Chromatographic determination of the mycotoxin patulin in fruit and fruit juices. J. Chromatogr. A 2000, 882 (1-2), 17-22. 22. M. Solfrizzo; G. Avantaggiato; A. Visconti, Use of various clean-up procedures for the analysis of ochratoxin A in cereals. J. Chromatogr. A 1998, 815 (1), 67-73. 23. A. Pfohl-Leszkowicz; R. A. Manderville, Ochratoxin A: An overview on toxicity and carcinogenicity in animals and humans. Mol. Nutr. Food Res. 2007, 51 (1), 61-99. 24. G. J. Soleas; J. Yan; D. M. Goldberg, Assay of Ochratoxin A in Wine and Beer by High-Pressure Liquid Chromatography Photodiode Array and Gas Chromatography Mass Selective Detection. J. Agric. Food. Chem. 2001, 49 (6), 2733-2740. 25. H. Reinhard; B. Zimmerli, Reversed-phase liquid chromatographic behavior of the mycotoxins citrinin and ochratoxin A. J. Chromatogr. A 1999, 862 (2), 147-159. 26. 行政院衛生署衛署食字第 0980462647 號,食品中真菌毒素限量標準,2009。 27. European Commission (EU). Commission Regulation (EC) No. 165/2010 of 26 February 2010 setting maximum levels for certain contaminants in foodstuffs. 28. European Commission (EU). Commission Regulation (EC) No. 1881/2006 of 19 December 2006 setting maximum levels for certain contaminants in foodstuffs. 29. G. J.; A. E., Validation of analytical methods for determining mycotoxins in foodstuffs. TrAC-Trends Anal. Chem. 2002, 21, 468-486. 30. D. R. Katerere; S. Stockenström; G. S. Shephard, HPLC-DAD method for the determination of patulin in dried apple rings. Food Control 2008, 19 (4), 389-392. 31. M. J. Barreira; P. C. Alvito; C. M. M. Almeida, Occurrence of patulin in apple-based-foods in Portugal. Food Chem. 2010, 121 (3), 653-658. 32. A. Desmarchelier; J.-M. Oberson; P. Tella; E. Gremaud; W. Seefelder; P. Mottier, Development and Comparison of Two Multiresidue Methods for the Analysis of 17 Mycotoxins in Cereals by Liquid Chromatography Electrospray Ionization Tandem Mass Spectrometry. J. Agric. Food. Chem. 2010, 58 (13), 7510-7519. 33. R. Rasmussen; I. Storm; P. Rasmussen; J. Smedsgaard; K. Nielsen, Multi-mycotoxin analysis of maize silage by LC-MS/MS. Anal. Bioanal. Chem. 2010, 397 (2), 765-776. 34. M. Sulyok; R. Krska; R. Schuhmacher, A liquid chromatography/tandem mass spectrometric multi-mycotoxin method for the quantification of 87 analytes and its application to semi-quantitative screening of moldy food samples. Anal. Bioanal. Chem. 2007, 389 (5), 1505-1523. 35. E. M. S. M. Gaspar; A. F. F. Lucena, Improved HPLC methodology for food control - furfurals and patulin as markers of quality. Food Chem. 2009, 114 (4), 1576-1582. 36. A. Molinié V. Faucet; M. Castegnaro; A. Pfohl-Leszkowicz, Analysis of some breakfast cereals on the French market for their contents of ochratoxin A, citrinin and fumonisin B1: development of a method for simultaneous extraction of ochratoxin A and citrinin. Food Chem. 2005, 92 (3), 391-400. 37. D. Spadaro; A. Ciavorella; S. Frati; A. Garibaldi; M. L. Gullino, Incidence and level of patulin contamination in pure and mixed apple juices marketed in Italy. Food Control 2007, 18 (9), 1098-1102. 38. S. C. Cunha; J. O. Fernandes, Development and validation of a method based on a QuEChERS procedure and heart-cutting GC-MS for determination of five mycotoxins in cereal products. J. Sep. Sci. 2010, 33 (4-5), 600-609. 39. E. Beltrán; M. Ibáñez; J. V. Sancho; F. Hernández, Determination of mycotoxins in different food commodities by ultra-high-pressure liquid chromatography coupled to triple quadrupole mass spectrometry. Rapid Comm. Mass Spectrom. 2009, 23 (12), 1801-1809. 40. G. Boonzaaijer; W. van Osenbruggen; A. Kleinnijenhuis; W. van Dongen, An exploratory investigation of several mycotoxins and their natural occurrence in flavour ingredients and spices, using a multi-mycotoxin LC-MS/MS method. World Mycotoxin J. 2008, 1 (2), 167-174. 41. S. Monbaliu; C. Van Poucke; C. Van Peteghem; K. Van Poucke; K. Heungens; S. De Saeger, Development of a multi-mycotoxin liquid chromatography/tandem mass spectrometry method for sweet pepper analysis. Rapid Comm. Mass Spectrom. 2009, 23 (1), 3-11. 42. A. Santini; R. Ferracane; G. Meca; A. Ritieni, Comparison and improvement of the existing methods for the determination of aflatoxins in human serum by LC-MS/MS. Anal. Methods 2010, 2 (7), 884-889. 43. V. Sewram; J. J. Nair; T. W. Nieuwoudt; N. L. Leggott; G. S. Shephard, Determination of patulin in apple juice by high-performance liquid chromatography-atmospheric pressure chemical ionization mass spectrometry. J. Chromatogr. A 2000, 897 (1-2), 365-374. 44. M. C. Spanjer; P. M. Rensen; J. M. Scholten, LC–MS/MS multi-method for mycotoxins after single extraction, with validation data for peanut, pistachio, wheat, maize, cornflakes, raisins and figs. Food Addit. Contam. Part A-Chem. 2008, 25 (4), 472 - 489. 45. P. Martos; W. Thompson; G. Diaz, Multiresidue mycotoxin analysis in wheat, barley, oats, rye and maize grain by high-performance liquid chromatography-tandem mass spectrometry. World Mycotoxin J. 2010, 3 (3), 205-223. 46. 李茂榮,質譜分析技術專輯,行政院國家科學委員精密儀器發展中心編印,2002,第五章,87-111。 47. 陳銘華,液相層析質譜術於食品中黃麴毒素及赭麴毒素之檢測-國立中興大學化學研究所碩士論文,2004。 48. M. C. García, The effect of the mobile phase additives on sensitivity in the analysis of peptides and proteins by high-performance liquid chromatography-electrospray mass spectrometry. J. Chromatogr. B 2005, 825 (2), 111-123. 49. J. T. Watson; O. D. Sparkman, Introduction to mass spectrometry:Instrumentation, Applications, and Strategies for Data Interpretation, John Wiley and Sons Inc., 2007, 486. 50. M. Anastassiades; S. J. Lehotay; D. Stajnbaher; F. J. Schenck, Fast and easy multiresidue method employing acetonitrile extraction/partitioning and “dispersive solid-phase extraction”for the determination of pesticide residues in produce. J. AOAC Int. 2003, 86 (2), 412-431. 51. D. Herebian; S. Zühlke; M. Lamshöft; M. Spiteller, Multi-mycotoxin analysis in complex biological matrices using LC-ESI/MS: Experimental study using triple stage quadrupole and LTQ-Orbitrap. J. Sep. Sci. 2009, 32 (7), 939-948.
摘要: 本實驗以QuEChERS萃取穀類產品中真菌毒素,再以LC/MS/MS進行分離及偵測,使用C18管柱以梯度沖提之方式進行分離,25分鐘內成功分離8種分析物。樣品處理部分,則探討不同之萃取溶劑種類、溶劑中添加酸之種類與比例、吸附劑之種類及添加量與添加鹽類比例,並對此方法進行線性範圍、線性相關係數、偵測極限、精密度與回收率之研究,最後將此方法應用於真實樣品中真菌毒素之分析。 由實驗結果顯示,以QuEChERS萃取技術結合液相層析質譜技術,使用正、負離子電灑游離法將分析物離子化後,再以選擇反應偵測(Selected reaction monitoring, SRM)進行偵測,取0.8 g穀類產品粉末加入2.5 mL含1%甲酸之乙腈:水(40:60, v/v)萃取溶劑,用震盪機以750 rpm震盪三分鐘進行萃取,加入300 mg之氯化鈉和500 mg無水硫酸鎂於離心管中,以750 rpm震盪三分鐘後,再以4000 rpm離心五分鐘後取上層液至另一離心管,加入45 mg之C18作為吸附劑和50 mg 無水硫酸鎂去除雜質,將離心管以1400 rpm震盪三分鐘,再以14000 rpm轉速離心五分鐘後,將萃取液通過0.22 μm濾膜,取20 μL注入液相層析質譜儀分析。 由實驗結果顯示,黃麴毒素與赭麴毒素檢測線性範圍在1.5~100 ng/g之間,棒麴毒素於150~800 ng/g,橘黴素則為25~1000 ng/g,線性相關係數均0.992以上,偵測極限定義為訊號三倍雜訊比,分別介於0.4 ng/g(黃麴毒素B1)至28 ng/g(棒麴毒素)之間。同日間與異日間之相對標準偏差分別為1.6%至17.1%與5.8%至19.4%,回收率介於72.5~112.4%之間,使用此方法對於一些真實樣品進行偵測,可測得橘黴素在某傳統市場穀類產品之含量為433.4 ng/g。
QuEChERS combined with LC-MS/MS was used to determine mycotoxins in grain products. The 8 mycotoxins were separated on a Waters XTerraTM MS C18 column in 25 minutes. The optimum parameters of MS/MS for eight mycotoxins and conditions of QuEChERS preparation were also discussed. 0.8g of homogenous grain product was precisely weighted and put into a 15mL centrifuge tube. Subsequently, 2.5 mL of mixed extraction solvent (acetonitrile:H2O:formic acid/40:60:1 v/v/v) was added into the same tube. After vigorously shaking for 3 min, 300 mg of sodium chloride and 500 mg magnesium sulfate were added. The tube was shaken intensely for 3 min then centrifuged at 4000 rpm for 5 min. The supernatant was transferred to 1.5 mL centrifuge tube then 45 mg octadecyl bonded silica gel and 50 mg magnesium sulfate were added. After intensely shaking for 3 min then centrifuging at 14000 rpm for 5 min, the supernatant was passed through a 0.22 μm syringe filter, then the extract was analyzed by LC-MS/MS. The analytical linearity for aflatoxins, ochratoxins, citrinin and patulin was from 1.5 to 100 μg/kg, 25 to 1000 μg/kg and 150 to 800 μg/kg, respectively. The limits of detection were from 0.4 to 1.0 μg/kg for aflatoxins, 0.4 to 0.6 μg/kg for ochratoxins, 6 μg/kg for citrinin and 28 μg/kg for patulin. The relative standard deviations of intra-day and inter-day analysis were from 1.7% to 17.1% and from 5.8% to 19.4%. The analysis of trace mycotoxin in grain products were also determined, the concentration of citrinin was detected at 433.4 ng/g. The results demonstrate the suitability of QuEChERS combined with LC-MS/MS approach to analyze trace amount of mycotoxins in grain products.
URI: http://hdl.handle.net/11455/16867
其他識別: U0005-1507201110522100
文章連結: http://www.airitilibrary.com/Publication/alDetailedMesh1?DocID=U0005-1507201110522100
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