Microsoft Word 刘文卫二校修改出1803

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1 7 5 Vol. 7 No Journal of Food Safety and Quality May, 刘文卫 *, 朱鹏飞, 周闰, 凌霞 (, ) 摘要 : 目的 (malachite green, MG) (leucomalachite green, LMG)(crystal violet, CV)(leucocrystal violet, LCV)- (UPLC-MS/MS) 方法,, - (electrospray ionization, ESI+)(multiple reaction monitoring, MRM), 结果 MG LMG CV LCV, LCV, ( 50%) MG LMG CV LCV 4 0.2~5.0 ng/ml, 0.999, 0.1~0.2 μg/kg MG LMG CV 93.5%~109.2%, 5.4%~9.6% (n=6) 结论, MG LMG CV LCV 关键词 : ; ; - ; ; Simultaneous determination of malachite green, crystal violet and their metabolites residues in aquatic products by ultra performance liquid chromatography-tandem mass spectrometry LIU Wen-Wei *, ZHU Peng-Fei, ZHOU Run, LING Xia (Wuxi Center for Disease Control and Prevention, Wuxi , China) ABSTRACT: Objective To establish a method for simultaneous determination of malachite green (MG), leucomalachite green (LMG), crystal violet (CV) and leucocrystal violet (LCV) residues in aquatic products by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Methods The samples were extracted with acetonitrile, cleaned up with neutral alumina column, and then detected by UPLC-MS/MS with electrospray ionization (ESI+) mode and multiple reaction monitoring (MRM) mode. Isotope internal standards were used for quantification. Meanwhile the matrix effects were also observed. Results Different matrix effects existed in 4 kinds of analytes, especially in the LCV. The recovery of LCV was low (about 50%). MG, LMG, CV, and LCV had good linear relationships in the range of 0.2~5.0 ng/ml (r>0.999). The limits of detection (LOD) were 0.1~0.2 μg/kg. The recoveries of MG, LMG and CV were ranged from 93.5% to 109.2%. The relative standard deviations (RSDs) were 5.4%~9.6% (n=6). Conclusion The method is rapid, accurate and sensitive, which is suitable for the detection of MG, LMG, CV and LCV residues in aquatic products. * 通讯作者 :,, liuwwcdc@126.com *Corresponding author: LIU Wen-Wei, Associate Chief Technician, Wuxi Center for Disease Control and Prevention, No.499, Jincheng Road, Wuxi , China. liuwwcdc@126.com

2 KEY WORDS: malachite green; crystal violet; ultra performance liquid chromatography-tandem mass spectrometry; aquatic product; matrix effect 1 引言 (malachite green, MG)(crystal violet, CV), (leucomalachite green, LMG) (leucocrystal violet, LCV) MG CV [1,2] MG LMG CV LCV [3-5], MG CV, 2002 MG, [6] MG,,, [6], MG LMG CV LCV [7,8] [9,10] [11-13] [14],, -(UPLC-MS/MS) MG LMG CV LCV 2 材料与方法 2.1 仪器与试剂 Agilent 1290/AB 3200 Q TRAP - (, ); Sigma 3-30K ( Sigma ); Vortex-Genie 2 ( Scientific Industries ), AG 245 ( Mettler Toledo ), (60 mg/3 ml, ) (Merck ) (CNW )( ), (MG, : 98.0 %) (LMG, : 94.0 %) (CV, : 96.0 %)(LCV, : 99.0%), Dr. Ehrenstorfer ; (MG-D 5, : 99.0 %) (LMG-D 5, : 99.8%), Witega 2013~ 试验方法标准溶液的配制 MG LMG CV LCV 10 mg( 0.1 mg),, 10 ml, μg/ml, μg/ml μl, 10 ml, 100 ng/ml, 4 MG-D 5 LMG-D 5 10 mg( 0.1 mg),, 10 ml, μg/ml, μg/ml μl, 10 ml, 100 ng/ml, ng/ml, ng/ml, 1.0 ng/ml 样品前处理 (1) 500 g,,,, 20 5 g( 0.1 g), 50.0 μl, 25 ml,, 8000 r/min 10 min,, (2) 5 ml, 5 ml,, 4 ml,, 50, 1 ml, 0.22 μm, 仪器条件 (1): Agilent C 18 (2.1 mm 50 mm, 1.8 μm); A, B 5 mmol/l 0.1% ; : 0.25 ml/min; : 5 μl; : 30 : 0~0.5 min, 30%A; 0.5~1 min, 30%~95%A; 1~3 min, 95%A; 3~3.5 min, 95%~30% A; 3.5~5 min, 30% A (2) : (electrospray ionization, ESI+), (multiple reaction monitoring, MRM), 5500 V, 500, 25 psi, 70 psi, 70 psi, Medium

3 5, : 基质效应的考察 [15], (ME/%)=S 2 /S 1 100%, S 1, S 2 ME<100%, [16] 3 结果与分析 3.1 质谱条件的优化,, MRM, (declustering potential, DP) (collision energy, CE), 1, 3.5 min, MRM 方法的线性范围及检出限 UPLC-MS/MS,,,, 0.2~5.0 ng/ml, 3 (limit of detection, LOD) r 方法的精密度和准确度, μg/kg, 1.0 μg/kg, 6,, LCV, MG LMG CV 93.5%~109.2%, (RSD) 5.4%~9.6%, LCV 38.1%~50.3%, RSD 3.9%~6.8%, 基质效应的考察, LCV, () 40%, 50%, MG LMG CV,,, LCV, LCV LMG-D 5,, CV LCV LCV 3.5 样品测定 2013~ , 15 MG, 6 LMG, 5 MG LMG, CV LCV Table 1 表 1 各组分的质谱参数 Mass spectrometry parameters of analytes (m/z) (m/z) DP(V) CE(eV) MG LMG CV LCV MG-D 5 LMG-D

4 : 1(MG, R t 1.9 min); 2(LMG, R t 2.6 min); 3(D 5 -MG, R t 1.9 min); 4(D 5 -LMG, R t 2.6 min); 5(CV, R t 2.2 min); 6(LCV, R t 2.7 min) 1 MRM Fig. 1 MRM chromatograms of analytes Table 2 表 2 各组分的回归方程相关系数和检出限 Regression equations, correlation coefficients and limits of detection of analytes r (μg/kg) MG MG-D 5 Y=1.47X LMG LMG-D 5 Y=0.809X CV MG-D 5 Y=6.13X LCV LMG-D 5 Y=0.358X Table 3 表 3 各组分的回收率与相对标准偏差 (n=6) Spiked recoveries and relative standard deviations of analytes(n=6) 0.5 μg/kg 1.0 μg/kg 2.0 μg/kg (%) RSD(%) (%) RSD(%) (%) RSD(%) MG LMG CV LCV 结论 - MG LMG CV LCV, MG LMG CV LCV 参考文献 [1],,,.

5 5, : [J]., 2007, 37(1): Zhai YX, Guo YY, Geng X, et al. Advances in studies on metabolic mechanism and bio-toxicity of malachite green [J]. Period Ocean Univ China, 2007, 37(1): [2],,,. [J]., 2007, 41(4): Cai CP, Lin G, Liu ZC, et al. Studies on the metabolism and depletion principles of crystal violet in eel tissues [J]. Chin J Vet Drug, 2007, 41(4): [3] Majeed SA, Nambi KSN, Taju G, et al. Cytotoxicity, genotoxicity and oxidative stress of malachite green on the kidney and gill cell lines of freshwater air breathing fish Channa striata [J]. Environ Sci Pollut Res, 2014, 21: [4] Chu YL, Chimeddulam D, Sheen LY, et al. Probabilistic risk assessment of exposure to leucomalachite green residues from fish products [J]. Food Chem Toxicol, 2013, 62: [5] Wan H, Weng S, Liang L, et al. Evaluation of the developmental toxicity of leucomalachite green administered orally to rats [J]. Food Chem Toxicol, 2011, 49: [6]. [D]. :, Li XL. Optimization of the detection methods of malachite green and dietary exposure assessment [D]. Chongqing: Southwest University, [7] Shen YD, Deng XF, Xu ZL, et al. Simultaneous determination of malachite green, brilliant green and crystal violet in grass carp tissues by a broad-specificity indirect competitive enzyme-linked immunosorbent assay [J]. Anal Chim Acta, 2011, 707: [8] Oplatowska M, Connolly L, Stevenson P, et al. Development and validation of a fast monoclonal based disequilibrium enzyme-linked immunosorbent assay for the detection of triphenylmethane dyes and their metabolites in fish [J]. Anal Chim Acta, 2011, 698: [9] GB/T [S]. GB/T Determination of malachite green and gentian violet residues in fishery products-high performance liquid chromatography with fluorescence detector [S]. [10] Xie J, Peng T, Chen DD, et al. Determination of malachite green, crystal violet and their leuco-metabolites in fish by HPLC-VIS detection after immunoaffinity column clean-up [J]. J Chromatogr B, 2013, : [11] Huang P, Zhao P, Dai X, et al. Trace determination of antibacterial pharmaceuticals in fishes by microwave-assisted extraction and solid-phase purification combined with dispersive liquid-liquid microextraction followed by ultra-high performance liquid chromatography-tandem mass spectrometry [J]. J Chromatogr B, 2016, 1011: [12],,,. [J]., 2013, 32(2): Sun YC, Li CT, Du NN, et al. Determination of malachite green and leucomalachite green residues in water and sediment by ultra-performance liquid chromatography-tandem mass spectrometry [J]. J Instrum Anal, 2013, 32(2): [13] Hurtaud-Pessel D, Couëdor P, Verdon E, et al. Determination of residues of three triphenylmethane dyes and their metabolites(malachite green, leuco malachite green, crystal violet, leuco crystal violet, and brilliant green) in aquaculture products by LC/MS/MS: first action [J]. J AOAC Int, 2013, 96(5): [14],,,. [J]., 2015, 6(1): Liu MY, Xiao SS, Yu B, et al. Research progress of the residue determination of triphenymethane dyes in aquatic product [J]. J Food Saf Qual, 2015, 6(1): [15] Eeckhaut AV, Lanckmans K, Sarre S, et al. Validation of bioanalytical LC MS/MS assays: evaluation of matrix effects [J]. J Chromatogr B, 2009, 877: [16] Matuszewski BK, Constanzer ML, Chavez-Eng CM. Strategies for the assessment of matrix effect in quantitative bioanalytical methods based on HPLC-MS/MS [J]. Anal Chem, 2003, 75(13): 作者简介 ( 责任编辑 : 姚菲 ) 刘文卫, 副主任技师, 主要研究方向为理化检验技术 liuwwcdc@126.com

Microsoft Word _刘丽丽-二校

Microsoft Word _刘丽丽-二校 7 2 Vol. 7 No. 2 2016 2 Journal of Food Safety and Quality Feb., 2016 刘丽丽 *, 刘谦, 王宇 (, 071001) 摘要 : 目的, 方法, HLB, C 18, ESI 结果 0.1~10 μg/kg, 0.999, 0.1 0.2 10.0 μg/kg 3, 95.0%~103.2%, 3.72%~6.85%, 0.1

Microsoft Word 刘文卫 二校 修改出1803

Microsoft Word 刘文卫二校修改出1803