Microsoft Word 大喜终校稿

Transcription

1 7 9 Vol. 7 No Journal of Food Safety and Quality Sep., , 肖英平 2, 李玲 3, 曾庆坤 3 1,, 杨华 2*, 任大喜 (1., ; 2., ; 3., ; 4., ) 4* 摘要 : 目的 - 方法 , - 结果 0.05~0.25 µg/l 88.2%~109.2% 8.06%, 6.89% 结论,, 关键词 : - ; ; ; Determination of quinolone residues in raw Holstein and Buffalo milk by high performance liquid chromatography-tandem mass spectrometry XIAO Ying-Ping 1, 2, LI Ling 3, ZENG Qing-Kun 3, YANG Hua 1, 2*, REN Da-Xi 4* (1. Institute of Quality and Standard for Agro-products, Zhejiang Academy of Agricultural Sciences, Hangzhou , China; 2. State Key Laboratory Breeding Base for Zhejiang Sustainable Pest and Disease Control, Zhejiang Academy of Agricultural Sciences, Hangzhou , China; 3. Water Buffalo Institute, Chinese Academy of Agricultural Science, Nanning , China; 4. College of Animal Sciences, Zhejiang University, Hangzhou , China) ABSTRACT: Objective To establish a method for the determination of quinolone residues in raw Holstein and Buffalo milk in south China by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). Methods One hundred and twenty milk samples were collected from Holstein and Buffalo farms in Zhejiang and Guangxi, and the quinolones residues in raw milk were determined by HPLC-MS/MS. Results The LODs and recovery rates were 0.05~0.25 µg/l and 88.2%~109.2%, respectively. The detection rates of quinolone residues in raw Buffalo and Holstein milk were 8.06% and 6.89%. Conclusion This method is accurate and reliable for the 基金项目 : (2010DS ZM1608) Fund: Supported by State Key Laboratory Breeding Base for Zhejiang Sustainable Pest and Disease Control (2010DS ZM1608) * 通讯作者 :,,, yanghua806@hotmail.com,,,, dxren@zju.edu.cn *Corresponding author: YANG Hua, Senior Livestock Engineer, Institute of Quality and Standard for Agro-products, Zhejiang Academy of Agricultural Sciences, Hangzhou , China. yanghua806@hotmail.com REN Da-Xi, Associate Professor, College of Animal Sciences, Zhejiang University, Hangzhou , China. dxren@zju.edu.cn

2 9, : determination of quinolones residues in raw milk. Compared to the results of other parts and area, the quinolone residues in raw Holstein and Buffalo are in safety situation in south China. KEY WORDS: high performance liquid chromatography-tandem mass spectrometry; Holstein; Buffalo; quinolone 1 引言 (quinolones, QNs), 20 60,,,,, [1],, [2],,,,,,, [3],, TTC STOP [4,5],, [6] [7] [8] [9],, [10-12], [13],,, [14] - (high performance liquid chromatography-tandem mass spectrometry, HPLC-MS/MS), 2 材料与方法 2.1 试剂与材料 : (pipemidic acid) (marbofloxacin) (fleroxacin) (ofloxacin) (pefloxacin) (enoxacin) (norfloxacin) (ciprofloxacin) (enrofloxacin) (lomefloxacin) (danofloxacin) (orbifloxacin) (difloxacin) (sarafloxacin) (sparfloxacin) (cinoxacin) (oxolinic acid) (nalidixic acid) (flumequin) Dr. Ehrenstorfer GmbH (, ); (, ), 15, , 58, ml, -20, 2.2 仪器与设备 Agilent1290 ( Agilent ); QTRAP 6500 ( AB ); (T8 Basic, IKA ); (945616): TALBOYS ; (Biofuge Primor): Thermo Scientific ; (HB10 digital): IKA ; (KQ-100B): 2.3 试验方法 标准溶液的配制 10 mg 50 ml,, 0.2 mg/ml 100 μl 200 ml, -0.1% (1:9, V:V), 100 ng/ml -0.1% 10 ng/ml, 样品的前处理 [15] 37, 2 g 50 ml, 50 µl 10 ml 4 ml, 1 min, 7000 r/min 3 min, 5 ml, 1 min, 4 ml 50 ml, 1 min, 7000 r/min 3 min, 1 min,,, 50

3 r/min, 1 ml ( :0.1% =1:9, V:V), 1 min,, 检测条件 ACQITY UPLC BEH C 18 (100 mm 2.1 mm, 1.7 μm), (A) 0.1% 2 mmol/l (B), 1 : 0.20 ml/min, 30, 10 µl : (electronic spray ion, ESI + ); 5500 V; 450 ; 50 psi; 60 psi 表 1 梯度洗脱程序 Table 1 The gradient elution procedure (min) (ml/min) A(%) B(%) 检测限 回收率及精密度试验 µg/l,, µg/l 3, 6, MultiQuant 3.0, 3 结果与分析 3.1 喹诺酮标准品 ~7.07 min,, 表 2 19 种喹诺酮类抗生素标准品保留时间和质谱信息 Table 2 Retention time and mass fragment ions of 19 kinds of QNs (min) (m/z) (V) (v) / / / / / / / / / / / / / / / / / / / Fig. 1 Chromatograms of 19 kinds of QNs

4 第9期 肖英平, 等: 高效液相色谱-三重四级杆质谱联用法检测荷斯坦和水牛原料乳中喹诺酮残留 续图 1 19 种喹诺酮标准品的色谱图 Fig. 1 Chromatograms of 19 kinds of QNs 3729

5 回收率 检出限和精密度 - 3 3, 19, (r 2 ) 0.993~0.999, 0.05~0.25 µg/l, [16,17], -, 0.3~2.0 ng/g 88.2%~109.2%, 70%~120%, Noemí [16] 96%~104.5%, 6 6.1%, -, 3.3 原料牛乳和水牛乳中喹诺酮含量 , 9, 5 ( 8.06%), 4 ( 6.89%), 0.06~1.02 μg/l 表 4 原料乳中喹诺酮检测结果 Table 4 Results of quinolone in raw milk μg/l 3/ ~1.02 2/ ~0.51 4/ ~0.87 2/ ~0.55 表 3 高效液相色谱串联质谱法测定喹诺酮抗生素的回收率 检出限及精确度 Table 3 The recovery rate, LOD and precision of quinolone residues determined by HPLC-MS/MS 1/ / ~0.62 2/ ~0.89 (%) (%) (µg/l) (%, RSD) 3/ ~0.35 2/ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ Noemí [16] 28, 5, 0.2~0.6 ng/g Chung [4] 269, 4 [18] - 30, 0.05 μg/l,, 4 结论 ~025 μg/l, 88.2%~109.7%, 6.1% 120, 8.06%, 0.06~1.02 μg/l; 6.89%, 0.06~0.89 μg/l - 参考文献 [1] Jiang W, Beier RC, Wang Z, et al. Simultaneous screening analysis of 3-methyl-quinoxaline-2- carboxylic acid and quinoxaline-2-carboxylic

6 9, : acid residues in edible animal tissues by a competitive indirect immunoassay [J]. J Agric Food Chem, 2013, 61(42): [2] Wang J, Leung D, Lenz SP. Determination of five macrolide antibiotic residues in raw milk using liquid chromatography-electrospray ionization tandem mass spectrometry [J]. J Agric Food Chem, 2006, 54, [3] Choma I, Grenda D, Malinowska I, et al. Determination of flumequine and doxycycline in milk by a simple thin-layer chromatographic method [J]. J Chromatogr B, 1999, 734: 7 14 [4] Chung H, Lee J, Chung Y, et al. Analysis of sulfonamide and quinolone antibiotic residues in Korean milk using microbial assays and high performance liquid chromatography [J]. Food Chem, 2009, 113: [5] Cho BH, Kim BH, Sohn SW, et al. Development of modified 2,3,5-triphenyltetrazolium chloride (TTC) reduction test for the detection of sulfonamide residues in raw milk [J]. Korean J Vet Public Health, 1993, 17(1): [6] Vidal JL, Aguileraluiz MM, Romerogonzález R, et al. Multiclass analysis of antibiotic residues in honey by ultraperformance liquid chromatography-tandem mass spectrometry [J]. J Agric Food Chem, 2009, 57(5): [7] Hu W, Xin L, He G, et al. Sensitive competitive immunoassay of multiple mycotoxins with non-fouling antigen microarray[j]. Bios Bioelect, 2013, 50(24): [8] Ediage EN, Mavungu JDD, Goryacheva IY, et al. Multiplex flow-through China Cattle Sci, 2008, 34(2): [14],,. [J]., 2001, 17(6): Li SY, Meng CL, Quan HH. The investigation report of antibiotic residues in pork, chicken and milk in Liu Zhou market[j]. Guangxi J Anim Husb Vet Med, 2001, 17(6): [15] Cho HJ, Yi H, Cho SM, et al. Single-step extraction followed by LC for determination of (fluoro) quinolone drug residues in muscle, eggs, and milk [J]. J Sep Sci, 2010, 33(8): [16] Noemi D, Alexandra Z, Alberto, et al. Simultaneous determination of quinolone and beta-lactam residues in raw cow milk samples using ultrasound-assisted extraction and dispersive-spe prior to UHPLC-MS/MS analysis [J]. Food Control, 2016, 60: [17] Jiang W, Beloglazova N, Wang Z, et al. Development of a multiplex flow-through immunoaffinity chromatography test for the on-site screening of 14 sulfonamide and 13 quinolone residues in milk [J]. Bios Bioelect, 2015, 66: [18],,,. - [J]., 2016, 25(1): 5 9. Yan Z, Ren MW, Zhang LB, et al. Simultaneous analysis of sulfonamides and quinolones in milk and milk powder by UFLC-MS/MS[J]. Chem Anal Meter, 2016, 25(1): 5 9. ( 责任编辑 : 杨翠娜 ) immunoassay formats for screening of mycotoxins in a variety of food matrices [J]. Anal Bioanal Chem, 2012, 403(1): [9] Ma X, Truong PL, Anh NH, et al. Single gold nanoplasmonic sensor for 作者简介 clinical cancer diagnosis based on specific interaction between nucleic acids and protein[j]. Bios Bioelect, 2014, 67(8): [10],. [J]., 2009, 30(8): Wang X, Che ZM, Huang TR. Assay on residue of antibiotics in pure milk 肖英平, 博士, 助理研究员, 主要研究方向为畜产品质量安全 ypxiaozju@126.com from market[j]. Food Res Dev, 2009, 30(8): [11],,,. - [J]., 2007, 40(5): Rao Y, Zeng ZL, Yang GX et al. Confirmation of Fluoroquinolone Residues in Milk by Liquid Chromatography-tandem Mass Spectrometry[J]. Sci Agric Sin, 2007, 40(5): 杨华, 高级畜牧师, 副院长, 主要研究方向为畜产品质量安全 yanghua806@hotmail.com [12] Antonio H, Javier H, Miguel R, et al. Determination of quinolone residues in infant and young children powdered milk combining solid-phase extraction and ultra-performance liquid chromatography-tandem mass spectrometry [J]. J Chromatogr A, 2011, 1218(42): [13],. [J]., 2008, 任大喜, 博士, 副教授, 博士生导师, 主要研究方向为营养与乳制品加工 dxren@zju.edu.cn 34(2): Li HZ, Cai SS. The detection of residues of antibiotics in cow s milk[j].