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1 5 3 Vol. 5 No Journal of Food Safety and Quality Mar., * 郑荣, 毛丹, 王少敏, 张甦, 季申 (, ) 摘要 : 目的 G 2 G 1 B 2 B 1 A B 1 B 2 T-2 10 方法 70%,, C 18, (multi-reaction monitoring, MRM), 结果 10, 3 60 ~100, RSDs(relative standard deviation) 0.4%~3.7% 结论, 关键词 : ; ; - Determination of 10 mycotoxins in peach kernel by high performance liquid chromatography-tandem mass spectrometry ZHENG Rong, MAO Dan, WANG Shao-Min, ZHANG Su, JI Shen * (Shanghai Institute for Food and Drug Control, Shanghai , China) ABSTRACT: Objective To establish a high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for determination of 10 kinds of mycotoxins (aflatoxing 2, G 1, B 2 and B 1, ochratoxin A, deoxynivalenol, zearalenone, fumonisinsb 1, B 2, and T-2 toxin) in peach kernel. Methods After extracted with 70% methonal and purified with solid-phase extraction column, 10 kinds of mycotoxins were analyzed by HPLC-MS/MS with multi-reaction monitoring (MRM) mode. Results These mycotoxins showed a good linear relationship. The mean recoveries at three spiked concentration levels were in the range of 60 ~100 with the relative standard deviation(rsds) from 0.4% to 3.7%. Conclusion This method is simple and accurate, and can satisfy the requirement of multi-mycotoxin analysis in Chinese herbal medicine. KEY WORDS: Peach kernel; multi-mycotoxin; high performance liquid chromatography-tandem mass spectrometry (mycotoxin),, 200,,, [1-3] * 通讯作者 :,, jishen2013@163.com *Corresponding author: JI Shen, Chief Physician, Shanghai Institute for Food and Drug Control, Shanghai ,China. jishen2013@163.com

2 3, : , 4, 5 [4] [5-8] A [9],,,,, ( 2),,,, [10-14], G2 G1 B2 B1 A B1 B2 T 材料与方法 1.1 仪器与试剂 ( Waters, Oasis HLB, 3 cc, 60 mg); - ( Agilent Table 1 表 1 国内外真菌毒素监测品种与限量规定 Specification and limits of mycotoxins in different nation standards G2 G1 B2 B1 5 B1 5 μg/kg, G2 G1 B2 B1 10 μg/kg G2 G1 B2 B1 G2 G1 B2 B1; A B1 G2 G1 B2 B1 G2 G1 B2 B1; A; ; ; ; T 2 ; B1 2 μg/kg, G2 G1 B2 B1 4 μg/kg 19 B1 10 μg/kg 8 B1 5 μg/kg, G2 G1 B2 B1 10 μg/kg (GB ) 表 2 真菌毒素的检测方法 Table 2 Detection methods of mycotoxins 4 HPLC 4 (1) (2) (3) (1)TLC;, (2)HPLC B1 A HPLC 4 HPLC (4 ) A T 2 (1) (2) HPLC/HPLC-MS

3 826 5, 1200 API5500, (electrospray ionization,esi); (N-EVAPTM112, Organomation Associates, Inc ) ( SUPELCO, : LB80734, G2 G1 B2 B1 0.3 µg/ml 1.0 µg/ml 0.3 µg/ml 1.0 µg/ml), A( Sigma, : L21764), ( FLuka, : SZE8150XV), ( FLuka, : SZE8150X), B1 B2( FLuka, : SZBC101XV, B1 B2 50 µg/ml), T-2 (, : LHAOM26),, H : B1 B2 G1 G2 A B1 B2 T-2, 1 ml 5 μg, ;, 1 ml 400 μg, 1.2 供试品溶液的制备 5 g(),, 70% 50 ml, 30 min, 5 ( 4000 r/min), 10 ml, 20 ml, 3 ml, ( 3 ml ),, ; 3 ml,,, 45, 50 1 ml, 1.3 液相色谱与质谱条件 : Phenomenex Kinetex XB-C 18 (1.7 μm, 100 mm, 2.1 mm); : 40 ; : 0.6 ml/min; : 5 µl; : - (1: 1) A, 0.01% B, 0.6 ml/min; 3, (ESI), G2 G1 B2 B1 B1 B2 T-2, A, 4 1 表 3 流动相梯度 Table 3 Mobile phase gradient /min A /% B /% 表 4 质谱参数表 Table 4 LC/MS/MS parameters /v /m/z /v /m/z /v B / / B / / G / / G / / B / / B / / T / / A / / / / / /

4 3, : Fig. 1 TIC spectrum in positive mode 2 Fig. 2 TIC spectrum in negative mode

5 结果 2.1 线性关系的考察,, 回收率试验 5 g(),, 18, 6, 3 ( B1 B2 G1 G2 A B1 B2 T ng/ml )0.25 ml 0.5 ml 1.0mL,,, 重复性试验 5 g(), 6,, 1 ml,,,, 7 Table 5 表 5 线性方程及相关系数 Regress equations and correlation coefficients /(ng ml -1 ) B1 0.2~4 Y=6040X B2 0.12~2.4 YY=3280 X G1 0.2~4 Y=4580 X G2 0.12~2.4 Y=1610 X B 1 1.6~32 Y=2590 X B 2 1.6~32 Y=5260 X T 2 1.6~32 Y=1610 X A 0.2~4 Y =2290 X ~1600 Y=4.88 X ~8 Y=3390 X 表 6 Table 6 回收率试验结果 Results of recovery / (μg kg 1 ) /% (n 6)/ B B G G B B T A

6 3, : 表 7 重复性试验结果 Table 7 Results of repeatability test /pg (n 6)/% B B G G B B T A 稳定性试验, 6 h,,, 0~24 h, 2.5 检测限, 3: 1, 样品测定,,, ( 9), 3 B1, 1,, 3 讨论 3.1 分析方法的选择, [11-16],,,,,, 表 8 检测限 Table 8 Detection limits / (μg kg -1 ) / (μg kg -1 ) B1 0.3 B B2 0.8 T G1 0.4 A 0.5 G B Table 9 表 9 样品测定结果 detection results of samples /(μg kg -1 ) B2 B1 B1 ( ) ( ) ( ) ( ) ( ) ( ) ( ) ( ) ( ) 136.2

7 830 5,,,,,,,,,,,,,, 3.2 分析条件的优化, Phenomenex Kinetex XB-C18,, T-2, A 3.3 样品处理方法的选择 : 1 70, HLB ; 2 84 (Mycosep 226) ; 3 84 MycoSpin 400,, 1, 70,, HLB,, 25 ml, 15 min,, 3 ml, HLB 3.4 中药中真菌毒素及其他生物毒素检测体系的建议 真菌毒素及其他生物毒素测定方法研究体系的原则 ( ), ( 10),,,, 监测品种体系,,,, T 2 6,,, Table 10 表 10 易感染中药品种及相应毒素种类 Easily contaminated herbs and corresponding mycotoxins 12 B1 B2 G1 G2 M1; M1 B1, 4 7 A 15, 60 (DON); 15 B1 B2 T 2 / T 2, HT-2, /

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9 832 5 作者简介 郑荣, 女, 副主任药师, 主要研究方向为中药质量控制和安全性检测研究 zhengrongsifdc@163.com 季申, 女, 主任药师, 主要研究方向为中药质量控制和安全性检测研究 jishen2013@163.com 功能性食品微生物 专题征稿 PUFA 投稿方式 : jfoodsq@126.com 食品安全质量检测学报 编辑部