Microsoft Word 邢丽红-二校出3618,3619

Size: px

Start display at page:

Download "Microsoft Word 邢丽红-二校出3618,3619"

诚嵇
4 years ago
Views:

1 7 9 Vol. 7 No Journal of Food Safety and Quality Sep., , 2, 邢丽红 3 1, 2,, 付树林 3 1, 2,, 彭吉星 3 1, 2,, 孙伟红 3* 1, 2,, 郑关超 3 1, 2,, 李兆新 3, 翟毓秀 1, 2, 3* (1., ; 2., ; 3. ( ), ) 摘要 : 目的 - 方法 3,5- (DNSAH), 2-,,, 结果 DNSAH 0.2~20 μg/l, r 2 >0.999 DNSAH 0.50~5.00 μg/kg, 88.3%~100%, <15% DNSAH 0.20 μg/kg 结论, 关键词 : ; 3,5- ; ; - Determination of nifursol metabolites in aquatic products by high performance liquid chromatography-tandem mass spectrometry XING Li-Hong 1,2,3, FU Shu-Lin 1,2,3, PENG Ji-Xing 1,2,3, SUN Wei-Hong 1,2,3*, ZHENG Guan-Chao 1,2,3, LI Zhao-Xin 1,2,3, ZHAI Yu-Xiu 1,2,3* (1. Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao , China; 2. Key Laboratory of Testing and Evaluation for Aquatic Product Safety and Quality, Ministry of Agriculture, Qingdao , China; 3. Laboratory of Quality & Safety Risk Assessment for Aquatic Products (Qingdao), Ministry of Agriculture, Qingdao , China) ABSTRACT: Objective To establish a method for the determination of nifursol metabolites in aquatic products by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). Methods The residue of nifursol metabolite of 3,5-dinitrosalicylic acid hydrazide (DNSAH) in sample was hydrolyzed with HCl, and derivatized with nitrobenzaldehyde. Then the analyte was extracted with acetonitrile and then analyzed by HPLC-MS/MS with internal standard method. Electrospray ionization was applied and operated in the negative ion mode. Results The calibration curve had a good linearity between area and the concentration of DNSAH of 0.2~20 基金项目 : 2014 ( ) ( ) Fund: Supported by Program of Agricultural Industry Standards Development ( ) and Natioanl Natural Science Foundation of China ( ) * 通讯作者 :,,,. sunwh@ysfri.ac.cn,,. zhaiyx@ysfri.ac.cn *Corresponding author: SUN Wei-Hong, Senior Engineer, Doctor, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, No.106, Nanjing Road, Shinan District, Qingdao , China. sunwh@ysfri.ac.cn ZHAI Yu-Xiu, Researcher, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, No.106, Nanjing Road, Shinan District, Qingdao , China. zhaiyx@ysfri.ac.cn

2 μg/l with the correlation coefficient over The average recoveries from spiked sample at three concentrations of 0.50, 2.00 and 5.00 μg/kg were ranged from 88.3% to 100% with intra and inter batch relative standard deviations below 15%. The detection limit of DNSAH was 0.20 μg/kg. Conclusion The efficient and simple method can be used to identify and quantify the nifursol metabolites in aquatic products with satisfactory sensitivity and repeatability. KEY WORDS: nifursol; 3,5-dinitrosalicylic acid hydrazide; aquatic products; high performance liquid chromatographytandem mass spectrometry 1 引言 (nifursol),, [1,2],,,,, [3-6] 3,5- (3,5-dinitrosalicylic acid hydrazide, DNSAH) (5-nitro-2-furoic acid), /2002/EC 90/2377/EC Ⅳ [7,8] /181/EC, (minimum required performance limits, MRPL) 1 μg/kg [9], [10,11] - (LC-MS) [12,13] - (LC-MS/MS) [14-17] (ELISA) [18,19] [20-22],,, -,, 2 材料与方法 2.1 仪器和设备 LC20 ( ); ABI ( AB ); ( Thermo Fisher ); ( ); ( Talboys ); (14000 r/min)( Sigma ); N-EVAP 112 ( Organomation ); IS-RDS3 ( ); Milli-Q ( Milipore ) 2.2 材料和试剂 3,5- (DNSAH) DNSAH- 15 N 2 ( Toronto Research Chemicals(TRC) ); 2- ( Sigma ), (, Merck ), ( ), (, ) : DNSAH DNSAH- 15 N 2, 1 ml,, 1.0 mg/ml, 试验方法,, DNSAH 2- (2-NBA) 2 Fig. 1 1 The molecular structures of nifursol and its metabolites

3 9, : Fig. 2 2 DNSAH Derived reaction mechanism of DNSAH with 2-NBA 水解和衍生化 2 g 50 ml, 50 μl, 15 min, 5 ml 0.5 mol/l HCl, 150 μl 50 mmol/l 2-, 1 min, h 提取和净化 1 mol/l, ph 7.0~ ml, 10 min, r/min 10 min 1 ml 2 ml, r/min 5 min, 0.22 μm, LC-MS/MS 标准工作曲线的制作 50 ml, DNSAH ng,, , 2.3.4, 液相色谱 - 串联质谱条件 (1) : Waters XTerra C 18 (2.1 mm 150 mm, 3.5 μm), : A.2 mmol/l, B., : 0~4 min, 90%A~10%A; 4~5 min, 10%A; 5~6 min, 10%A~90%A; 6~7 min, 90%A; : 0.3 ml/min; : 30 ; : 10 μl (2) (ESI), : V; : 550, (CAD): Medium; (curtain gas): 30 psi; (Gas1): 35 psi; (Gas2): 35 psi; (DP): -110V; : (MRM); DNSAH 1 3 结果与分析 3.1 色谱条件优化 NP-DNSAH, %,,, NP-DNSAH,,,,,, 质谱条件的优化 10 ng DNSAH, 2.3, NP-DNSAH NP-DNSAH- 15 N 2,, NP-DNSAH NP-DNSAH- 15 N 2,,, 1 表 1 Table 1 选择反应监测母离子子离子和碰撞能量 Parent ions and product ions of DNSAH and the collision energy (m/z) DNSAH ESI DNSAH- 15 N 2 ESI : * (m/z) (ev) 183* *

4 Fig. 3 3 Chromatograms of DNSAH before and after optimization 3.3 前处理条件优化衍生化实验条件的研究 1 DNSAH, , 4 Fig.5 5 The relationship curve between derivatization time and derivative product Fig. 4 4 The relationship curve between temperature and derivative product, DNSAH 16 h, 37, 25, 60,,, 37 2, DNSAH 37, DNSAH 5 5, 10 h,, 14~18 h,,, 16 h,,,, 18 h, 16 h ph 值对提取率的影响 ph 3.0~10.0, 6 6, DNSAH ph, ph 3.0~4.0, ph, DNSAH, ph 6.5,, ph7.5,, ph 3.0~4.0, DNSAH, ph,,, ph 6.0,, ph,,, ph 7.0~7.5

5 9, : Fig. 6 6 ph DNSAH Effects of ph on extraction rate of DNSAH 提取试剂的选择 [23,24],, DNSAH,, DNSAH ( 2), DNSAH ( 7),,,, 8 Table 2 表 2 不同定容溶剂对测定结果的影响 Effects of different constant volume solvents on the determination results A B C D E F G DNSAH : A. :2 mmol/l =5:95; B. :2 mmol/l =20:80; C. :2 mmol/l =40:60; D. :2 mmol/l =50:50; E. :2 mmol/l =60:40; F. :2 mmol/l =80:20; G. :2 mmol/l =95:5 7 DNSAH Fig. 7 Effects of matrix on DNSAH

6 方法评价线性关系和检出限 DNSAH ng,,, DNSAH 3 DNSAH 0.999, 0.20~20 μg/l ; 3 DNSAH 0.20 μg/kg, μg/kg 方法回收率精密度, 3, 6, 3,, 4, 0.50~5.00 μg/kg 88.3%~100%, 5.07%~10.8%, 6.60%~9.00%, DNSAH 8 表 3 硝呋索尔代谢物 DNSAH 的线性范围线性方程相关系数 Table 3 Linear range, linear equation and correlation coefficient of nifursol metabolite DNSAH (μg/l) (r) DNSAH 0.20~20 Y= X 表 4 样品中 DNSAH 的添加回收率和精密度数据表 (n=6) Table 4 The recovery and precision of DNSAH in the samples(n=6) (μg/kg) (μg/kg) (%) RSD(%) RSD(%) , 0.486, , 98.8, , 5.07, , 1.83, , 96.9, , 8.55, , 5.24, , 99.3, , 9.47, , 0.487, , 97.7, , 6.21, , 2.05, , 94.8, , 9.01, , 5.32, , 97.7, , 9.88, , 0.433, , 89.3, , 9.68, , 2.12, , 93.0, , 8.17, , 5.32, , 95.7, , 7.35, Fig. 8 8 DNSAH (A. ; B. ; C. ) Characteristic ion mass chromatograms of DNSAH(A.DNSAH standard; B. carp blank sample; C. carp spiked sample)

7 9, : 结论, 2-,,, DNSAH,,,, 参考文献 [1] Eric V, Pierrick C, Pascal S. Multi-residue monitoring for the simultaneous determination of five nitrofurans (furazolidone, furaltadone, ni trofurazone, nitrofurantoine, nifursol) in poultry muscle tissue through the detection of their five major metabolites (AOZ, AMOZ, SEM, AHD, DNSAH) by liquid chromatography coupled to electrospray tandem mass spectrometry In-house validation in line with Commission Decision 657/2002/EC [J]. Anal Chim Acta, 2007, 586: [2],,. [J]., 2010, 31(2): Zhu WX, Liu YF, Liang W. Progress on nitrofuran drug residues in animal-borne food [J]. Prog Vet Med, 2010, 31(2): [3] Zuidema T, Mulder PPJ, van Rhijn JA, et al. Metabolism and depletion of nifursol in broilers [J]. Anal Chim Acta, 2005, 529: [4] Vahl M. Analysis of nifursol residues in turkey and chicken meat using liquid chromatography- tandem mass spectrometry [J]. Food Addit Contam, 2005, 22(2): [5] Mulder PPJ, Zuidema T, Keestra NGM, et al. Determination of nifursol metabolites in poultry muscle and liver tissue. Development and validation of a confirmatory method [J]. The Analyst, 2005, 130: [6],,. [J]., 2014, 4(6): Wang Q, Lv HY, Song Y. Research progress of nitrofurans elimination law in aquatic products and aquaculture environment [J]. Chin Fish Qual Stand, 2014, 4(6): [7] Council Regulation No. 1756/2002/EC [Z]. Official Journal of the European Communities, 2002, L 265/1. [8] Council Regulation No. 2377/1990/EC [Z]. Official Journal of the European Communities, 1990, L 224/1. [9] Commission Decision No. 2003/181/EC [Z]. Official Journal of the European Communities, 2003, L 71/17. [10],,,. 7 [J]., 2011, 47(5): Zhang HC, Li J, Yan XD, et al. Determination of seven nitrofuran drugs in feeds by a high performance liquid chromatography method [J]. Chin J Vet Med, 2011, 47(5): [11] [S]. Announcement of the Ministry of Agriculture Determination of nitrofurans in feeds-high performance liquid chromatography [S]. [12] Finzi JK, Donato JL, Sucupira M, et al. Determination of nitrofuran metabolites in poultry muscle and eggs by liquid chromatography-tandem mass spectrometry [J]. J Chromatogr B, 2005, 824(1 2): [13] Laure T, Olivier P, Guy D, et al. Advantages of LC-MS-MS compared to LC-MS for the determination of nitrofuran residues in honey [J]. Anal Bioanal Chem, 2006, 386(7 8): [14] Zhang YB, Qiao HO, Chen C, et al. Determination of nitrofurans metabolites residues in aquatic products by ultra-performance liquid chromatography tandem mass spectrometry [J]. Food Chem, 2016, 192: [15] Kaufmann A, Butcher P, Maden K, et al. Determination of nitrofuran and chloramphenicol residues by high resolution mass spectrometry versus tandem quadrupole mass spectrometry [J]. Anal Chim Acta, 2015, 862: [16] Dongkyum K, Byungjoo K, Seok-Won H, et al. An optimized method for the accurate determination of nitrofurans in chicken meat using isotope dilution liquid chromatography/mass spectrometry [J]. J Food Compos Anal, 2015, 41: [17],,,. - [J]., 2011, 32(4): Zhang LJ, Li JJ, Zhang LT, et al. Determination of 4 nitrofurans and chloramphenicol in fisheries water by liquid chromatography coupled tandem mass spectrometry [J]. J Chin Mass Spectr Soc, 2011, 32(4): [18],. ELISA( cielisa) [J]., 2005, 35(2): Luo J, Li J. An Indirect Competitive ELISA Method for the Quantitative Analysis of Furazolidone [J]. Period Ocean Univ China, 2005, 35(2): [19] Franek M, Diblikova I, Vass M, et al. Validation of a monoclonal antibody-based ELISA for the quantification of the furazolidone metabolite(aoz)in eggs using various sample preparation [J]. Vet Med, 2006, 51(5): [20] Jorge B, Andreia F, Sara M, et al. Detection, accumulation, distribution, and depletion of furaltadone and nifursol residues in poultry muscle, liver, and gizzard [J]. J Agric Food Chem, 2011, 59(2): [21],,,. [J]., 2012, 31(11): Zhao FJ, Xiao CG, Yue ZF, et al. Determination of metabolite residue of nifursol in animal tissue by high performance liquid chromatographytandem mass spectrometry [J]. J Instrum Anal, 2012, 31(11): [22],,,. SPE-LC-MS/MS 5 [J]., 2013, 29(6): Hou XC, Huang JF, Du ZF, et al. Quantitative determination of five nitrofuran metabolites in meat meal using solid phase extraction and LC-tandem MS[J]. Mod Food Sci Technol, 2013, 29(6): [23] [S]. Announcement of the Ministry of Agriculture Determination of nitrofuran metabolic residues in aquatic products by LC-MS/MS method[s]. [24] GB/T / [S].

3622 7 GB/T 21311-2007 Determination of residues of

8 GB/T Determination of residues of nitrofuran metabolites in foodstuffs of animal origin HPLC-MS/MS method [S]. ( 责任编辑 : 白洪健 ) 孙伟红, 高级工程师, 主要研究方向为水产品质量安全及风险评估研究 sunwh@ysfri.ac.cn 作者简介邢丽红, 助理研究员, 主要研究方向为水产品质量安全 xinglh@ysfri.ac.cn 翟毓秀, 研究员, 主要研究方向为水产品质量安全 zhaiyx@ysfri.ac.cn

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

Microsoft Word 邢丽红-二校 出3618,3619

Microsoft Word 邢丽红-二校出3618,3619