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1 6 10 Vol. 6 No Journal of Food Safety and Quality Oct., 2015 刘印平 1, 路杨 1, 王丽英 1, 常凤启 1 1,, 杨立新 2, 张永茂 (1.,, ; 2., ) 1* 摘要 : 目的 - (liquid chromatography-tandem mass spectrometry, LC-MS/MS) 3 (FB 1, FB 2 and FB 3 ) 方法 50% /,, /, 13 C 34 -FBs 结果 75.9%~108.2%, 0.39~0.58 µg/kg, 67% 3 (FB 1, FB 2 and FB 3 ) 结论,, 关键词 : ; ; - ; ; Determination of fumonisins in corn by liquid chromatography-tandem mass spectrometry with isotope dilution method LIU Yin-Ping 1, LU Yang 1, WANG Li-Ying 1, CHANG Feng-Qi 1, YANG Li-Xin 1, 2, ZHANG Yong-Mao 1* (1. Hebei Provincial Center for Disease Control and Prevention, Hebei Provincial Key Laboratory of Food Safety Risk Monitoring, Shijiazhuang , China; 2. College of Environmental Sciences and Engineering, Peking University, Beijing , China) ABSTRACT: Objective To establish a simple and rapid method for the determination of fumonisin (FB 1, FB 2 and FB 3 ) in corn by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Methods Homogenized samples were sequentially extracted by 50% acetonitrile-water, centrifuge, multifunction columns cleanup, evaporate, re-dissolve and filtration before analysis. Quantification of fumonisin was based on the use of 13 C 34 -fumonisin. Results The present method showed adding standard recoveries of 75.9%~108.2%, with limit of detections (LODs) of FB 1, FB 2 and FB 3 ranging from 0.39 to 0.58 µg/kg. Analysis of 18 corn samples by the current method was successfully performed. The simultaneous occurrence of FB 1, FB 2 and FB 3 was observed in 67% of samples. Conclusion This quantitative method is demonstrated to be rapid, simple, and high sensitive, which can be applied in the accurate determination of fumonisin in corn. KEY WORDS: fumonisin; corn; liquid chromatography-tandem mass spectrometry; isotope dilution; multifunction columns * 通讯作者 :,, @163.com *Corresponding author: ZHANG Yong-Mao, Senior Technical Personnel, Hebei Provincial Center for Disease Control and Prevention, No.97, Huai an East Road, Shijiazhuang , China @163.com

2 引言, 3 : B1(FB 1 ) B2(FB 2 ) B 3 (FB 3 ), [1-3],,, [4,5] FB 1 FB 1,,, FB 1 FB 2 FB [6-10] , (FDA) (2 mg/kg) (1~50 mg/kg) 1 mg/kg,, (immuno-affinity chromatography, IAC) [11-13], (MultiSep 211 FUM),,,, - 13 C 34 -FBs,, 2 材料与方法 2.1 试剂与仪器 - (TSQ, ); TDL-5-A ( ); AE200 ( METTLER ); Atlantis T3 (15 mm 2.1 mm, 1.8 μm) (Waters, Ireland); MultiSep 211 FUM Romer Labs (Tulln, Austria) : FB 1 FB 2 FB 3 (50 µg/ml) 13 C 34 -FB 1 (25 µg/ml) 13 C 34 -FB 2 (10 µg/ml) 13 C 34 -FB 3 (10 µg/ml) Sigma-Aldrich Laborchemikalien GmbH (Steinheim, Germany) (, Fisher ); (, ) Milli-Q (Millipore, Bedford, MA, USA) 2.2 样品前处理 2.0 g 50 ml, ( 13 C-FB 1, 13 C-FB 2, 13 C-FB 3), 10 ml - (5:5, V:V), 3 min, 4000 r/min 10 min( 4 ), 10 ml, - (3:1, V:V) 5 ml - (5:5, V:V) 5 ml, 10 ml / (99:1, V:V), 1 ml - (5:5, V:V) 0.22 µm - (liquid chromatography-tandem mass spectrometry, LC-MS/ MS) 2.3 液相色谱条件 : Atlantis T3 (15 mm 2.1 mm, 1.8 μm) : A: 0.1% ; B: / (5:5, V:V) 1, : 0.2 ml/min; : 25 ; : 20 μl Table 1 表 1 液相色谱梯度洗脱条件 Gradient elution of liquid chromatography (min) A(%) B(%) 串接质谱仪操作条件 (ESI+); : 3.5 kv; : 100 ; (, ) : 320 ; : 30 V; : 600 L/h; 60 L/h; 40 kpa, 6 kpa,

3 10, : : Ar 2.5 标准储备液和标准工作溶液的配制, 50%,, ( ng/ml), 5 ng/ml 3 结果与讨论 3.1 质谱条件优化 2, 方法的线性范围和检出限, ng/ml, 3, 3,, 0.995, 1.4~1.9 µg/kg 3.3 回收率和精密度, µg/kg 4, 6, 4, 75.9%~108.2%, (RSDs) 2.1%~9.4%,, 3.4 实际样品测定 18, 16 FB 1,, 4.8~486 µg/kg 12 FB 2, 1.9~42.3 µg/kg 14 FB 3, 2.2~111.6 µg/kg, 12 FB 1 FB 2 FB 3 Wang [14] FB 1, 96.1% FB 1, 20~6568 µg/kg, 2878 µg/kg 表 2 LC-MS/MS 测定伏马菌素的主要参考质谱参数 Table 2 Parameters for the LC-MS/MS analysis of fumonisins (RT, min) (m/z) (m/z) /ev FB FB FB C 34 -FB C 34 -FB C 34 -FB

4 Fig. 1 1 (ESI+, 5 ng/ml) The chromatogram of fumonisins standard solution (5 ng/ml, ESI+) Table 3 表 3 线性回归方程和方法检出限, 定量限 Equation of linear regression, limit of detection (LOD, S/N=3), limit of quantification (LOQ, S/N=10) and linearity (r 2 ) of fumonisins FBs r 2 LOD(µg/kg) LOQ(µg/kg) FB 1 Y=0.0362X FB 2 Y=0.0541X FB 3 Y=0.0491X Table 4 表 4 LC/ESI(+)-MS/MS 方法测定伏马菌素的准确度和精密度 Accuracy and precision of the LC/ESI(+)-MS/MS method for total fumonisins FBs ±RSD(%, n=6) 2 µg/kg 10 µg/kg 100 µg/kg 500 µg/kg FB ± ± ± ±2.1 FB ± ± ± ±2.9 FB ± ± ± ±3.3

10, : - 3821 4 结论与讨论 - 3,,,,,, 300,,, 参考文献 [1] Hartl M, Humpf HU. Toxicity assessment of fumonisins using the brine shrimp (Artemia salina) bopassau [J]. Food Chem Toxicol, 2000, 38: 1097 1102.

[3] Gumprecht LA, Smith GW, Constable PC, et al. Species and organ specificity of fumonisin-induced endothelial alterations: potential role in porcine pulmonary edema [J].

5 10, : 结论与讨论 - 3,,,,,, 300,,, 参考文献 [1] Hartl M, Humpf HU. Toxicity assessment of fumonisins using the brine shrimp (Artemia salina) bopassau [J]. Food Chem Toxicol, 2000, 38: [2] Kwon OS, Slikker JW, Dacies DL. Biochemical and morphological effects of fumonisin B1 on primary cultures of fat cerebrum [J]. Neurotoxcicol Teratol, 2000, 22: [3] Gumprecht LA, Smith GW, Constable PC, et al. Species and organ specificity of fumonisin-induced endothelial alterations: potential role in porcine pulmonary edema [J]. Toxicology, 2001, 160, 1 3, 71. [4] Yoshizawa T, Yamashita A, Luo Y. Fumonisin occurrence in corn from high- and low-risk areas for human esophageal cancer in China [J]. Appl Environ Microbiol, 1994, 60: [5] Shephard CS, Snijman PW. Elimination and excretion of a single dose of the mycotoxin fumonisin B2 in a non-human primate [J]. Food Chem Toxic, 1999, 37: [6] Kedera CJ, Plattner RD, Desjardins AE. Incidence of Fusarium spp. and levels of fumonisin B1 in maize in western Kenya [J]. Appl Environ Microbiol, 1999, 65: [7] Omurtag GZ. Determination of fumonisin B1 and B2 in corn and corn-based products in Turkey by high performance liquid chromatography [J]. J Food Prot, 2001, 64: [8] Gutema T, Munimbazi C, Bullerman LB. Occurrence of fumonisins and moniliformin in corn and corn-based food products of U.S. origin [J]. J Food Prot, 2000, 63: [9] Seo E, Yoon Y, Kim K, et al. Fumonisins B1 and B2 in agricultural products consumed in South Korea: an exposure assessment [J]. J Food Prot, 2009, 72: [10] Caldas ED, Silva AC. Mycotoxins in corn-based food products consumed in Brazil: an exposure assessment for fumonisins [J]. J Agric Food Chem, 2007, 55: [11] Visconti A, Solfrizzo M, De Girolamo A. Determination of fumonisins B1 and B2 in corn and corn flakes by liquid chromatography with immunoaffinity column cleanup: collaborative study [J]. J AOAC Int, 2001, 84: [12] Solfrizzo M, De Girolamo A, Gambacorta L, et al. Determination of fumonisins B1 and B2 in corn-based foods for infants and young children by LC with immunoaffinity column cleanup: interlaboratory validation study [J]. J AOAC Int, 2011, 94: [13] Brera C, Debegnach F, Minardi V, et al. Immunoaffinity column cleanup with liquid chromatography for determination of aflatoxin B1 in corn samples: interlaboratory study [J]. J AOAC Int, 2007, 90: [14] Wang Y, Liu SL, Zheng H, et al. T-2 toxin, zearalenone and fumonisin B1 in feedstuffs from China [J]. Food Add Contam: Part B, 2013, 6: 作者简介 ( 责任编辑 : 杨翠娜 ) 刘印平, 硕士, 中级技师, 主要研究方向为食品安全检测 liuyinping0930@163.com 张永茂, 副主任技师, 主要研究方向为预防医学与食品安全检测 @163.com

DOI /j.issn Food Research And Development UPLC-MS/MS 5 0.

DOI /j.issn Food Research And Development UPLC-MS/MS 5 0. 118 217 5 38 1 DOI 1.3969/j.issn.5-6521.217.1.28 Food Research And Development 1 2 1 2 1 2 1 2 1 2 1 2 1. 731 2. 731 - UPLC-MS/MS 5. 1 mol/l ProElut PXC 5 % - Thermo Hypersil Gold mm 2.1 mm 1.9 μm ESI