Microsoft Word 谢丹 二校 出2105,2107,2108

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1 7 5 Vol. 7 No Journal of Food Safety and Quality May, 2016 谢 丹 1, 邓春丽 1 2,, 赵云峰 3, 吴永宁 2, 3* (1., ; 2., ; 3., ) 摘要 : 目的 方法 - -, (ZEN) (DON), 结果 ZEN DON 3~500 ng/ml, 0.999, 95.05%~107.10%,ZEN DON 0.03 μg/kg 0.9 μg/kg ZEN DON 0.1μg/kg 3μg/kg 30, DON, ZEN 100%, 333 μg/kg, 1.95 μg/kg, 67.7 µg/kg, 400 μg/kg 结论, 关键词 : ; ; ; - Investigation of the contamination status of zearalenone and deoxynivalenol in edible oil XIE Dan 1, DENG Chun-Li 1, ZHAO Yun-Feng 2, 3 2, 3*, WU Yong-Ning (1. College of Food Science and Engineering Wuhan Polytechnic University, Wuhan , China; 2. China National Central for Food Safety and Risk Assessment, Beijing , China; 3. Key Laboratory of Food Safety Risk Assessment Ministry of Health, Beijing , China) ABSTRACT: Objective To analyze the pollution status of zearalenone and deoxynivalenol in edible oil sold in Beijing market. Methods Zearalenone(ZEN), deoxynivalenol(don) in the collected samples including peanut oil, corn oil, blend oil and other edible oils were detected and analyze by the method of isotope dilution-ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) under the multiple reaction monitoring mode. Results Zearalenone(ZEN), deoxynivalenol(don) had a good linear relationship in the range of 3~500 ng/ml with the correlation coefficients of and the recoveries were 95.05% ~ %. The detection limits of ZEN and DON were 0.03 μg/kg and 0.9 μg/kg and the quantification limits of ZEN and DON were 0.1 μg/kg and 3 μg/kg. The results of 30 samples collected from Beijing market showed that, DON was not detected, the detection rate of ZEN was 100% with the highest content of 333 μg/kg and the lowest content of 1.95 μg/kg, while 基金项目 : ( ) Fund: Supported by The National Science Fund For Distinguished Young Scholars( ) * 通讯作者 :,,, wuyongning@cfsa.net.cn *Corresponding author: WU Yongning, Professor, China National Central for Food Safety and Risk Assessment, NO.37, Guangqu Road, Chaoyang Distract, Beijing , China wuyongning@cfsa.net.cn

2 the mean amount was 67.7 μg/kg, which was lower than the tolerance limits specified in EU. Conclusion By the analysis of the toxin content in edible oils we could get a preliminary understanding of the pollution of ZEN and DON in edible oil in Beijing. KEY WORDS: zearalenone; deoxynivalenol; vegetable oil; ultra performance liquid chromatography-tandem mass spectrometry 1 引言 (zearalenone, ZEN) F-2,,, [1-4], ZEN, (EFSA) ZEN ZEN, 2006 ZEN 400 μg/kg [5] ZEN, GB ZEN 60 µg/kg [6] (deoxynivalenol, DON),,,, [7] DON, DON,, [8, 9] DON, GB [10] 1000 µg/kg ZEN DON :, [11], ZEN 69.30%, DON 84.65% [12,13] ZEN DON, ZEN DON, DON 97.4%, ZEN 87.6%, ZEN DON,,,, - - (TLC) (ELISA) (HPLC) - (LC-MS/MS) TLC ELISA, ; HPLC,, ; UPLC-MS/MS,,, 2 材料与方法 2.1 仪器与试剂 (, ); (, CNW ); : MycoSep226 ( Romer Labs ); Milli-Q : (ZEN, 61.6 μg/ml) 13 C- ( 13 C-ZEN, 10.1 μg/ml) 1 ZEN DON Fig. 1 Structural formula of ZEN and DON, (DON, μg/ml) 13 C- ( 13 C-DON, 10.1 μg/ml), Romer Labs ; : ZEN(53~105 μg/kg, Romer Labs ), DON(0.7~0.9 ppm, Romer Labs ); Waters UPLC I-Class-Xevo TQ-S - ( ); Mettler Toledo ( - ( ) ); QB-210

3 5, : 2107 ( ); SIGMA ( SIGMA ); (Eppendorf Reasch plus, Eppendorf ); ( Corning ); ; 2.2 油样, 30 ( 1), 4 表 1 样品信息 Table 1 Sample information ( ) % 36% 6% 2%

4 实验方法 溶液的配制 : 10 μl ZEN (61.6 μg/ml) 20 μl DON (100.1 μg/ml) -0.2% -10 mmol/l (V:V, 10:90) 1 ml, ZEN ng/ml, DON 2000 ng/ml; : -0.2% -10 mmol/l (V:V, 10:90), ( 13 C-ZEN 13 C-DON), ZEN ng/ml, DON ng/ml, 13 C-ZEN 3.01 ng/ml, 13 C-DON 10.1 ng/ml 样品前处理 (1) 2 g( ±0.02 g) 15 ml, 100 μl ( 13 C-ZEN 3.01 ng/ml, 13 C-DON 10ng/ml,), 10 ml : (V:V, 86:14), 1 h, 1 h, 9000 r/min 15 min, (2) 8 ml MycoSep 226, 4 ml, 40 1 ml -0.2% 10 mm (V:V, 10:90) 30 s, 0.20 μm, 仪器条件 (1) UPLC A: ; B: 0.2% +10 mmol/l ; : 0.3 ml/min; 2; : 40 C; : 10 µl (2) : ESI; : 0.5 kv; : 150 ; : 150 L/h; : 500 ; : 1000 L/h; : (multiple reaction monitoring, MRM), 3 3 结果与讨论 3.1 方法的线性范围与定量限 2.3.3,,,,, 4 4, 4 3~500 ng/ml, S/N=10 (limit of quantitation, LOQ) S/N=3 (limit of detection, LOD) Table 2 表 2 UPLC 梯度洗脱程序 Gradient elution program for UPLC (min) (ml/min) A(%) B(%)

5 5, : 2109 表 3 质谱参数 Table 3 MS parameters (m/z) (V) (m/z) (ev) (m/z) (ev) DON C-DON ZEN C-ZEN Table 3 表 4 线性范围与定量限 Linear equations and the limit of quantitation (μg/kg) ZEN Y=0.6734X ~600 ng/ml DON Y =1.0413X ~500 ng/ml 回收率与质控样 2, ZEN %, DON 95.05%; 2, ZEN DON 2g, ZEN 54.1 μg/kg, DON μg/ml, 3.3 样品和标准品的色谱图 2, 2, ZEN 162 μg/kg, DON, ZEN 8.81 min, DON 3.68 min 植物油中真菌毒素污染调查分析, 30 DON, ZEN 100%, ZEN 1.95 μg/kg, 333 μg/kg, 67.7 μg/kg, EFSA 400 μg/kg; DON 0% ZEN 5, 8, 100~400 μg/kg, 6, 7 5 7, : > >, ZEN 4 讨论 [14], ZEN, ZEN μg/kg 333 μg/kg, 148 μg/kg, [15] ZEN ; [16], ZEN,, ZEN DON, ZEN, ZEN DON, 3 [17] ( ), DON, ;, DON, :,, [18],,, [19]

6 Fig. 2 MRM chromatograms of positive samples

7 5, : Fig. 3 MRM chromatograms of standards

8 表 5 植物油样品中 ZEN 检测结果 Table 5 Results of ZEN in edible oils ( ) (μg/kg) (μg/kg) (μg/kg) (μg/kg) 表 6 植物油样品中 DON 检测结果 Table 6 Results of DON in edible oils ( ) (μg/kg) (μg/kg) (μg/kg) (μg/kg) 6 <LOD <LOD <LOD <LOD 11 <LOD <LOD <LOD <LOD 12 <LOD <LOD <LOD <LOD 1 <LOD <LOD <LOD <LOD 表 7 含量偏高样品的检测结果 Table 7 Results of samples with high content (μg/kg) ZEN(μg/kg) ,,, 参考文献 [1],,,. [J]., 2012, (9): Liu YZ, Zhu FH, Ning XJ, et al. Research progress on pollution and toxicity of zearalenone in feed [J]. Feed Rev, 2012, (9): [2],, [J]., 2011, 23(2): Jiang HZ, Yang WR, Yang ZB. Metabolism toxicity and preventive measures of zearalenone [J]. Chin J Anim Nutr, 2011, 23(2): [3],,,. -HPLC [J]., 2013, 2: Bao L, Jing P, Xu YL, et al. Determination of zearalenone in edible oil by HPLC-fluorometry with immunoaffinity column clean up [J]. Acad Peri Farm Prod Proc, 2013, 2: [4],. [J]., 2007, 28(2): Deng YT, Yuan H. The development of zearalenone toxicity mechanism [J]. Progr Veterin Med, 2007, 28(2): [5] Drzymala SS, Weiz S, Heinze J, et al. Automated solid-phase extraction coupled online with HPLC-FLD for the quantification of zearalenone in edible oil [J]. Anal Bioanal Chem, 2015, (407): [6] GB [S]. GB Hygienic standard for grains [S]. [7] TUTELYAN V A. Deoxynivalenol in cereals in Russia [J].Toxico Lett, 2004, 153(1): [8],,,. [J]., 2012, 24(1): Yin J, Wu L, Peng ZX, et al. Deoxynivolenol: toxicity and mechanisms of action [J]. Chin J Anim Nutr, 2012, 24(1): [9],,,. B 1 B 2 [J]., 2015, 51(4): Cui XN, Li H, Wang HT, et al. Detemination of zearaleimne and its metabolites and fumonisins B l and B 2 in the grain and feed with

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