Microsoft Word 张璐琪-二校

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1 7 5 Vol. 7 No Journal of Food Safety and Quality May, β- 张璐琪 1*, 王凤美 2, 张鸿伟 2, 徐彪 2, 梁成珠 (1., ; 2., ) 2 摘要 : 目的 3 - 方法 (CLB) (RAC) (SAL)3 β-, - (LC-MS/MS) 结果 - - LC-MS/MS 83.3 % 87.5%(CLB) 12.5 %(RAC) 100%(SAL), CLB RAC SAL β- ELISA - 94 % 94% 76.2 % 75 % 结论,,, 关键词 : β- ; ; ; Detection of 3 kinds of β-agonists in animal foods by rapid test card and ELISA kit ZHANG Lu-Qi 1*, WANG Feng-Mei 2, ZHANG Hong-Wei 2, XU Biao 2, LIANG Cheng-Zhu 2 (1. College of Food Science and Engineering, Ocean university of China, Qingdao , China; 2. Technical Center for Inspection and Quarantine of Shandong Entry-Exit Inspection and Quarantine Bureau, Qingdao , China) ABSTRACT: Objective To evaluate the accuracy of the rapid test card and enzyme-linked immunosorbent assay (ELISA) kit in detecting 3 kinds of β-agonists in animal foods. Method Three kinds of β-agonists residues including clenbuterol (CLB), ractopamine (RAC) and salbutamol (SAL) in animal foods were detected by rapid test card and ELISA, respectively. The results were compared with those of the liquid chromatography-tandem mass spectrometry (LC-MS/MS) and the methods were evaluated. Results The coincidence rate of clenbuterol rapid test card and LC-MS/MS was 83.3%. The coincidence rates of triple rapid test card and LC-MS/MS were 87.5%, 12.5% and 100% for CLB, RAC and SAL, respectively. The coincidence rates of ELISA kits and LC-MS/MS were 94%, 94%, 76.2% and 75% for CLB, RAC, SAL and β-agonists, respectively. Conclusion The accuracy of kits are relatively high, but the false positive and false negative of some kits are obvious. While the rapid test card has the high LOD and low accuracy, it does not meet the general detecting demand. KEY WORDS: β-agonists; rapid test card; enzyme-linked immunosorbent assay; accuracy evaluation * 通讯作者 :,, @163.com *Corresponding author: ZHANG Lu-Qi, Master, College of Food Science and Engineering, Ocean University of China, Qingdao , China @163.com

2 5, : 3 β 引言 β- ( ) (clenruberol, CLB) (ractopamine, RAC) (salbutamol, SAL),,,,, [1] β-,,, [2], CLB RAC SAL (HPLC) [3-5] - (GC-MS) [6,7] (ELISA) [8,9] [10,11], [12] ELISA ELISA, - (LC-MS/MS), 2 材料与方法 2.1 材料与仪器 LC-MS/MS ( 0.05 μg/kg) (ph=3); (, Merck ) ( : CLB 3 μg/kg RAC 3 μg/kg SAL 5 μg/kg, ); ELISA ( : 0.04 μg/kg, r-biopharm, 12445); ELISA ( : 0.35 μg/kg 0.2 μg/kg,, ); β- ELISA ( : 0.1 μg/kg,, CS1506) TECAN infinite M200 ( Tecan ), Himac CR22GII ( HITACHI ),,,,, MS1Minishaker ( IKA ) 2.2 检测方法速测卡 (1) : 3 g ; 95 10~15 min; r/min, min, (2):, 2 ( 40 μl), 30 s 1~2 ; 5 min, 盐酸克仑特罗试剂盒 (1) : 5 g, 25 ml 50 mmol/l HCl, 20 min; 8000 r/min, min; 18 ml (3 g ), 2 ml 0.5 mmol/l NaOH, 10 min; 10 ml 500 mmol/l KH 2 PO 4 (ph=3), h; 8000 r/min, min; 10 ml (1 g ), (20~25 ), RIDA C 18 (2) RIDA C 18 : 3 ml (100 %) ; 2 ml 50 mmol/l KH 2 PO 4 (ph=3); ; 2 ml 50 mmol/l KH 2 PO 4 (ph=3) ; 2 min ; 1 ml (100 %), 15 /min, ; 50~60 ; 1 ml, 20 μl (3):, ; 100 μl, 15 min;, 250 μl,, 2,, ; 20 μl, 100 μl,, 30 min;, 250 μl,,,, ; 100 μl /,, 15 min; 100 μl, 30 min 450 nm 莱克多巴胺和沙丁胺醇 ELISA 试剂盒 (1) : 2 g, 4 ml ( 20 1:20 ), 2 ml 0.1 mol/l, 30 s; 8000 r/min 10 min; 1 ml, 1 mol/l ph 6.75±0.25,, 8000 r/min 5 min, ; 50 μl (2):, ; 50 μl, 100 μl,, 10 min( 30 min);, 250 μl,, 3,,

3 ; 100 μl /,, 10 min; 100 μl, 10 min 450 nm β- 兴奋剂 ELISA 试剂盒 (1) : 4 g, 8 ml, 30 min; 8000 r/min 10 min, 4 ml, ; 1 ml ( 10 ), 2 ml, 30 s, 2000 r/min 10 min; 100 μl, 400 μl B,, 50 μl (2):, ; 50 μl, 50 μl 50 μl,, 30 min;, 250 μl, 3, ; 150 μl, 10 min; 50 μl, 450 nm 2.3 试验设计盐酸克伦特罗速测卡 5 clen ( μg/kg) 20, 盐酸克伦特罗 - 莱克多巴胺 - 沙丁胺醇三联速测卡 2 clen ( μg/kg) 2 rac ( μg/kg) 2 sal ( μg/kg) 20, 盐酸克伦特罗试剂盒 5 clen ( μg/kg) 20, 莱克多巴胺试剂盒 5 rac ( μg/kg) 20, 沙丁胺醇试剂盒 3 sal ( μg/kg) 18, β- 兴奋剂试剂盒 2 clen ( μg/kg) 2 sal ( μg/kg) 20, 2 3 结果与分析 3.1 金标速测卡 1,, 83.3%, (3~9 μg/kg),,,,, (7~9 μg/kg) 3.2 ELISA 试剂盒 2,, 94%, β-, 76.2 % 75 %, IC 50,, 3 IC 50,, β- 表 1 快速检测卡的检出结果 Table 1 Detection results of rapid test cards Clen Rac Sal 6/10 0/4 4/4 4/4 19/20 28/28 0/28 28/28 /% : ; ;

4 5, : 3 β 表 2 Table 2 ELISA 试剂盒的检出结果 Detection results of ELISA kits ELISA β- 8/10 7/10 6/6 8/8 39/40 40/40 26/36 28/40 /% : ; ; 表 3 各试剂盒 IC 50 及线性范围比较 (μg/kg) Table 3 Comparison of IC 50 and linear ranges of kits(μg/kg) ELISA β , , , , IC ~ ~ ~ ~20.00 : IC 50 50%, 4 讨论 β-,,,,,,,,,, : (1),, [13] (2), RIDA C 18 C 18,, ;,, [14] (3),,,, (4),, (5) [15], β-,,,, 5 结论,,,,,,,,,, β-,, 参考文献 [1],,. [J].

5 1962 7, 2008, 19: Liu WH, Li L, Qin L. Determination of clenbuterol hydrochloride in pork by enzyme-linked immunosorbent assay [J]. Anhui Agric Sci Bull, 2008, 19: [2],,,. [J]., 2008, 6: 61. Lu XC, Hou YZ, Wang HL, et al. Comparison of rapid test card and kit in detecting clenbuterol hydrochloride [J]. Shanghai J Anim Husb Vet Med, 2008, 6: 61. [3],,,. [J]., 2010, 6: Wu YL, Yang T, Shan JH, et al. Determination of residual clenbuterol enantiomers in swine urine by high performance liquid chromatography [J]. Chin J Anal Chem, 2010, 6: [4],. 6 β [J]., 2008, 7: Cai XX, Zhang XY. Rapid simultaneous determination of six β-agonists in food stuffs of animal origin by ultra-performance liquid chromatography coupled with triple quadrupole mass spectrometry [J]. Chin J Heal Lab Technol, 2008, 7: [5],,, β 2 - [J]., 2014, 10: Liu J, Xie YF, Ren DD, et al. Determination of residues of 26 β 2 -agonists in pork liver by reversed phase high performance liquid chromatographytandem mass spectrometry [J]. Chin J Anal Chem, 2014, 10: [6],,,. [J]., 2008, 12: , Zhao JJ, Li YL, Wang ZQ, et al. Investigation of determination of clenbuterol hydrochloride in hair of rat using capillary by GC-MS [J]. Chin J Heal Lab Technol, 2008, 12: , [7],,. -- [J]., 2002, 11: Xie MX, Liu Y, Jiang M. Determination of clenbuterol in meat sample by coupling solid phase extraction with gas chromatography-mass spectrometry [J]. Chin J Anal Chem, 2002, 11: [8]. ELISA [J]., 2014, 5: Liu DX. Comparison of determination of clenbuterol hydrochloride by ELISA, colloidal gold test paper and high performance liquid chromatography [J]. Chin J Health Lab Technol, 2014, 5: [9],,,. - [J]., 2015, 8: He FY, Yu XN, Wang K, et al. Determination of salbutamol residues in pork by ELISA and LC-MS/MS [J]. Chin Brew, 2015, 8: [10],,,. (ELISA) [J]., 2005, 11: Liu ZZ, Yin WZ, Du YP, et al. Rapid screening of clenbuterol residue in pig urine by enzyme-linked immunosorbent assay(elisa) combined with colloidal gold test paper [J]. Chin Anim Quarant, 2005, 11: [11],,,. β- [J]., 2015, 2: Xiong L, Li WH, Gao YQ, et al. A review of inspection technology of β-agonist drug residues in meat [J]. J Food Saf Qual, 2015, 2: [12],,,. [J]., 2013, 5: Ye XZ, Wang Q, Zhao SP, et al. Evaluation of rapid detection of clenbuterol hydrochloride in urine [J]. Zhejiang Agric Sci, 2013, 5: [13]. ELISE [J]., 2008, 6: Yu J. Application of ELISE on residue determination of clenbuterol hydrochloride [J]. Meat Ind, 2008, 6: [14],,. [J]., 2008, 1: Fan F, Mao AM, Ni GJ. Error analysis and solutions of the determination of clenbuterol hydrochloride by enzyme-linked immunosorbent assay[j]. J Anim Sci Vet Med, 2008, 1: [15]. [J]., 2015, 29: 115. Zha WY. Common influence factors and control methods of enzyme-linked immunosorbent assay [J]. World Latest Med Inf, 2015, 29: 115. 作者简介 ( 责任编辑 : 金延秋 ) 张璐琪, 硕士, 主要研究方向为食品安全检测 @163.com

Microsoft Word - 4839-4843 20150924002-李建煌-二校全出

Microsoft Word - 4839-4843 20150924002-李建煌-二校全出 6 12 Vol. 6 No. 12 215 12 Journal of Food Safety and Quality Dec., 215 ONPG 李建煌 1, 姚蕾 1, 李玉优 1, 李焕仪 1, 何亮辉 1, 姜春宝 2, 曹维强 (1., 51852; 2., 5166) 1* 摘要 : 目的 βd(onitrophenylβdgalactopyranoside,