欢迎访问作物学报,今天是

作物学报 ›› 2012, Vol. 38 ›› Issue (01): 186-189.doi: 10.3724/SP.J.1006.2012.00186

• 研究简报 • 上一篇    

高效液相色谱-紫外法同时检测小麦中DON、15ACDON和3ACDON

杨丹,耿志明,马鸿翔*,姚金保,张旭,张平平,张鹏   

  1. 江苏省农业科学院 / 江苏省农业生物学重点实验室,江苏南京 210014
  • 收稿日期:2011-05-27 修回日期:2011-09-18 出版日期:2012-01-12 网络出版日期:2011-09-30
  • 通讯作者: 马鸿翔, E-mail: mahx@jaas.ac.cn
  • 基金资助:

    本研究由江苏省农业科技自主创新基金项目[CX(09)635, CX(10)128], 现代农业产业技术体系项目(nycytx-03), 国家科技部国际合作项目(2009DFA32020)和国家转基因生物新品种培育重大专项(2008ZX08002-001)资助。

Establishment of a HPLC-UV Method for Simultaneous Determination of DON, 15ACDON, and 3ACDON in Wheat

YANG Dan,GENG Zhi-Ming,MA Hong-Xiang,YAO Jin-Bao,ZHANG Xu,ZHANG Ping-Ping,ZHANG Peng   

  1. Provincial Key Laboratory of Agrobiology / Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China
  • Received:2011-05-27 Revised:2011-09-18 Published:2012-01-12 Published online:2011-09-30
  • Contact: 马鸿翔, E-mail: mahx@jaas.ac.cn

摘要: 建立了测定小麦中B型单端孢酶烯族毒素脱氧雪腐镰刀菌烯醇(deoxynivalenol, DON)、3-乙酰脱氧雪腐镰刀菌烯醇(3-acetyldeoxynivalenol, 3-ACDON)和15-乙酰脱氧雪腐镰刀菌烯醇(15-acetyldeoxyni-valenol, 15-ACDON)的高效液相色谱-紫外(HPLC-UV)检测方法。用水提取小麦样品,提取液经无水乙醇等体积沉淀,再结合Oasis HLB固相萃取小柱可取得较好的净化效果。采用乙腈/0.005%磷酸水溶液二元梯度洗脱程序在高效液相色谱-紫外检测器上完成DON、15ACDON和3ACDON的定性定量分析。结果表明,在0.5~15.0 mg L-1线性范围内,DON、15ACDON和3ACDON的平均加标回收率分别为89.8%、93.4%和92.9%,相对标准偏差分别为2.2%、2.0%和2.5% (n=3);检测限分别为12.2、10.5和16.7 μg kg-1。该方法准确、重现性好,样品净化方法使杂峰干扰少,大幅减少有机溶剂的使用,成本低,适用于小麦中B型单端孢酶烯族毒素的大批量检测。

关键词: 小麦, B型单端孢酶烯族毒素, 高效液相色谱-紫外检测法

Abstract: A HPLC-UV method was established to determine deoxynivalenol (DON), 3-acetyldeoxynivalenol (3-ACDON) and 15- acetyldeoxynivalenol (15-ACDON) in wheat. Wheat sample was extracted by water, and the extracting solution was precipitated with an equal volume of ethanol followed by solid phase extraction with Oasis HLB cartridge. Qualitative and quantitative analyses of DON, 15ACDON and 3ACDON were done by HPLC-UVD using gradient elution program with acetonitrile and 0.005% phosphoric acid solution. Results showed that at the linear range of 0.5–15.0 mg L-1, the recoveries were 89.8%, 93.4%, and 92.9%, respectively. The relative standard deviations were 2.2%, 2.0%, and 2.5%, respectively, and the detection limit levels were 12.2, 10.5, and 16.7 μg kg-1, respectively. The method was accurate and reproducible. Sample purification lessened interference peaks, and due to the use of organic solvents was greatly reduced, the cost was decreased. Therefore, the method was suitable for the large batch determination of type B trichothecenes in wheat.

Key words: Wheat, B trichothecenes, HPLC-UV

[1]Cavaliere C, D’Ascenzo G, Foglia P, Pastorini E, Samperi R, Laganà A. Determination of type B trichothecenes and macrocyclic lactone mycotoxins in field contaminated maize. Food Chem, 2005, 92: 559–568
[2]Wang Y-Z(王裕中), Miller J D. Toxin producing potential of Fusarium graminearum from China. Mycosystema (真菌学报), 1994, 13(3): 229–234 (in Chinese with English abstract)
[3]Zhang P(张鹏), Ma H-X(马鸿翔). Biosynthesis and regulation of trichothecenes in Fusarium species. Biotechnol Bull (生物技术通报), 2009, 1: 11–15 (in Chinese with English abstract)
[4]FAO/WHO, Summary report of the seventy-second meeting of the Joint FAO/WHO Expert Committee on Food Additives (JECFA), issued 16th March 2010, available at http://www.who.int/foodsafety/chem/summary72 rev.pdf
[5]Wolf C E, Bullerman L B. Heat and pH alter the concentration of deoxynivalenol in an aqueous environment. Food Prot, 1998, 61: 365–367
[6]Wang Y-Z(王裕中), Chen H-G(陈怀古), Yang X-Y(杨新宁), Lu M(陆鸣), Wu Z-F(吴志凤). Bioactivity of crude toxin produced by Fusarium graminearum and its application in identification of fusarium head blight resistance of wheat cultivars. China Agric Sci (中国农业科学), 1989, 2(4): 54–57 (in Chinese with English abstract)
[7]Wei R-Y(魏润蕴). Thin-layer chromatography determining of deoxynivalenol in wheat. J Hyg Res (卫生研究), 1986, 15(5): 40–43 (in Chinese)
[8]Trucksess M W, Page S W, Wood G E, Cho T H. Determination of deoxynivalenol in white flour, whole wheat flour, and bran by solid-phase extraction/liquid chromatography: interlaboratory study. J AOAC Int, 1998, 81: 880–886
[9]Scott P M, Kanhere S R. Comparison of column phases for separation of derivatized trichothecenes by capillary gas chromatography. J Chromatogr, 1986, 368: 374–380
[10]Tacke B K, Casper H H. Determination of deoxynivalenol in wheat, barley, and malt by column cleanup and gas chromatography with electron capture detection. J AOAC Int, 1996, 79: 472–475
[11]Jestoi M, Ritieni A, Rizzo A. Analysis of the Fusarium mycotoxins fusaproliferin and trichothecenes in grains using gas chromatography-mass spectrometry. J Agric Food Chem, 2004, 52: 1464–1469
[12]Klötzel M, Gutsche B, Lauber U, Humpf H U. Determination of 12 type A and B trichothecenes in cereals by liquid chromatography electrospray ionization tandem mass spectrumetry. J Agric Food Chem, 2005, 53: 8904–8910
[13]Park J J, Chu F S. Assessment of mimunochemical methods for the analysis of trichothecene mycotoxins in naturally occur-ringmoldy corn. J AOAC Int, 1996, 79: 465–471
[14]Casale W L, Pestka J J, Hart L P. Enzyme-linked mimunosor-bentassay employingmonoclonalantibody specific for deoxynivalenol (vomitoxin) and sever alanalogues. J Agric Food Chem, 1988, 36: 663–668
[15]Laamanen I, Veijalainen P. Factors affecting the results of T-2 mycotoxin ELISA assay. J Food Addit Contam, 1992, 9: 337–343
[16]Turner N W, Subrahmanyam S, Piletsky S A. Analytical methods for determination of mycotoxins: a review. Anal Chim Acta, 2009, 632: 168–180
[17]Klötzel M, Schmidt S, Lauber U, Thielert G, Humpf H U. Comparison of different clean-up procedures for the analysis of deoxynivalenol in cereal-based food and validation of a reliable HPLC method. Chromatographia, 2005, 62: 41–48
[18]Jiménez M, Mateo R. Determination of mycotoxins produced by Fusarium isolates from banana fruits by capillary gas chromatography and highperformance liquid chromatography. J Chromatogr A, 1997, 778: 363–372
[19]Luongo D, Severino L, Bergamo P, D’Arienzo R, Rossi M. Trichothecenes NIV and DON modulate the maturation of murine dendritic cells. Toxicon, 2010, 55: 73–80
[20]Yasuaki D, Sayaka K, Masaomi K, Yukie S, Tomoaki H, Keisuke S, Kunitoshi M, Susumu K, Yoshiko S K, Makoto S. Rapid deposition of glomerular IgA in BALB/c mice by nivalenol and its modifying effect on high IgA strain (HIGA) mice. Exp Toxicol Pathol, 2011, 63: 17–24
[21]Bony S, Olivier-Loiseau L, Carcelen M, Devaux A. Genotoxic potential associated with low levels of the Fusarium mycotoxins nivalenol and fusarenon X in a human intestinal cell line. Toxicol in Vitro, 2007, 21: 457–465
[1] 翟胜男, 曹新有, 李豪圣, 李吉虎, 李法计, 刘金栋, 夏先春, 吕莹莹, 马瑞峰, 王颖, 耿洪伟, 刘建军. 小麦Pod-A1Pod-D1Pod-2D位点等位变异对籽粒过氧化物酶活性的遗传效应分析[J]. 作物学报, 2026, 52(6): 1593-1603.
[2] 习千辉, 徐梓瑗, 刘梦梦, 王宏艺, 郎凯琳, 井震海, 陈锋, 赵磊. 小麦籽粒铜含量的全基因组关联分析及候选基因预测[J]. 作物学报, 2026, 52(6): 1604-1617.
[3] 毛嘉琦, 黄朋雨, 赵佳佳, 郑兴卫, 武棒棒, 郝宇琼, 屈非, 刘成, 马朋涛, 郑军. 山西小麦品种白粉病抗性评价及抗病基因分子检测[J]. 作物学报, 2026, 52(6): 1669-1681.
[4] 胡川, 赵凯男, 黄修利, 吴金芝, 任开明, 王贺正, 付国占, 黄明, 李友军. 一次灌溉下耕作方式和氮肥用量对旱地小麦产量和品质的影响[J]. 作物学报, 2026, 52(6): 1830-1846.
[5] 陈雪燕, 何华川, 李政嘉, 董新盼, 李藕琪, 刘小云, 李丹萍, 陈志伟, 刘国霞, 吕胜源, 吴印莹, 赵振东, 曹新有, 万何平. 水培盐碱复合胁迫下‘济麦60’苗期根系有机酸分泌动态变化及其转录调控机制[J]. 作物学报, 2026, 52(6): 1859-1875.
[6] 高沛阳, 李瑾璇, 董宇奎, 石玉, 张振, 张永丽. 测墒补灌下小麦分蘖发生和成穗对施氮量的响应[J]. 作物学报, 2026, 52(6): 1847-1858.
[7] 张献丰, 郭利建, 李康春, 孔斌雪, 刘玉芳, 车卓, 杨德龙. 小麦ABHD6基因家族鉴定与粒重功能标记开发[J]. 作物学报, 2026, 52(6): 1711-1727.
[8] 王壮壮, 武紫君, 张永新, 张芯源, 袁丽雪, 陈如雪, 刘世举, 段剑钊, 冯伟, 王同朝, 王永华. 豫东南黏壤潮土区水氮优化协同提高冬小麦产量和氮素利用效率[J]. 作物学报, 2026, 52(5): 1501-1521.
[9] 何万龙, 耿洪伟, 张飞飞, 米克热阿依·阿巴白克热, 罗紫洋, 李鹏程, 周钊宇, 程宇坤. 基于深度学习的小麦重要病害图像识别系统的研究[J]. 作物学报, 2026, 52(5): 1401-1417.
[10] 张振, 冯连杰, 石玉, 于振文, 张永丽. 节水补灌下不同穗型小麦产量形成差异研究[J]. 作物学报, 2026, 52(5): 1522-1535.
[11] 侯思宇, 王国璀, 韦金贵, 谢玮欣, 殷文, 樊志龙, 柴强, 胡发龙. 绿肥配施化学氮肥对西北干旱灌区小麦干物质积累及产量形成的影响[J]. 作物学报, 2026, 52(4): 1208-1219.
[12] 尚云秋, 赵竹, 陈欢, 丁永刚, 乔玉强, 李玮, 张向前, 曹承富, 杜世州. 长期定位耕作方式对雨养小麦籽粒灌浆和产量形成的影响[J]. 作物学报, 2026, 52(4): 1236-1250.
[13] 乔宇馨, 李程越, 康晓玉, 张鑫琪, 贾绍辉, 刘倩, 曹亚丽, 史鑫蕊, 郝兴宇, 李萍. 基于APSIM模型的长期免耕秸秆覆盖对旱地小麦增产效应研究[J]. 作物学报, 2026, 52(4): 1181-1192.
[14] 李灿, 张喜伟, 朱博涛, 张沛沛. 小麦GSK激酶TaSK41的功能分析及互作蛋白的筛选[J]. 作物学报, 2026, 52(3): 677-687.
[15] 侯洁, 付朵朵, 武海峰, 郝宇琼, 郑兴卫, 武棒棒, 周凯, 李晓华, 郑军, 赵佳佳. 山西省小麦地方品种的染色体多样性及遗传效应分析[J]. 作物学报, 2026, 52(3): 746-763.
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
No Suggested Reading articles found!