6 12 Vol. 6 No. 12 2015 12 Journal of Food Safety and Quality Dec., 2015-14 * 吴健, 朱峰, 吉文亮, 李放, 阮丽萍, 刘华良 (, 210009) 摘要 : 目的 14 - 方法 0.22 μm, (0.05%)+ -, Atlantis dc-c 18, (electro spray ion source, ESI) (multi-reaction monitoring, MRM) 结果 14, 5 ng/l~1000 ng/l, 10 ng/l~1000 ng/l, 20 ng/l~1000 ng/l, 50 ng/l~1000 ng/l, r 13 0.99, 83.9%~110.0%, 2.4%~15.4%, 1.0 ng/l~20 ng/l 结论, 关键词 : ; ; ; - Simultaneous determination of 14 quinolone residues in water by high performance liquid chromatography-tandem mass spectrometry WU Jian, ZHU Feng, JI Wen-Liang, LI Fang, RUAN Li-Ping, LIU Hua-Liang * (Jiangsu Provincial Center for Disease Prevention and Control, Nanjing 210009, China) ABSTRACT: Objective To establish a rapid analytical method for the simultaneously screening and detection of 14 quinolone residues in water by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). Methods After a high speed centrifugation, the samples were filtered 0.22 µm millipore filter and then were separated on an Atlantis dc-c18 column with the mobile phases of formic acid (0.05%) and methanol-acetonitrile. The electrospray ionization (ESI) source in positive ion mode was used for multiple reaction monitoring (MRM). Results The 14 quinolone chemical components showed a good linear relationship in their concentration ranges with the correlation coefficients more than 0.99 except ciprofloxacin. The linear relationship of ofloxacin, enrofloxacin, lomefloxacin, difloxacin, sarafloxacin, flumequine and gatifloxacin were 5 ng/l~1000 ng/l, ciprofloxacin, orbifloxacin, marbofloxacin and pefloxacin were 10 基金项目 : (BE2010745) (ZX201109) Fund: Supported by Jiangsu Provincial Science and Technology Support Project (BE2010745) and Jiangsu Provincial 12th Five-Year Health Development Project with Science and Education (ZX201109) * 通讯作者 :,,, E-mail: LHL-1 @163.com *Corresponding author: LIU Hua-Liang, Associate Researcher, Jiangsu Provincial Center for Disease Control and Prevetion, No.172, Jiangsu Road, Nanjing 210009, China. E-mail: LHL-1 @163.com
12, : - 14 4967 ng/l~1000 ng/l, fleroxacin and enoxacin were 20 ng/l~1000 ng/l, and danofloxacin was 50 ng/l~1000 ng/l. The recoveries were all of 83.9%~110.0% and the relative standard deviation (RSD) were all of 2.4%~15.4%. The limits of detection (LOD) were all of 1.0 ng/l~20 ng/l. Conclusion This method is rapid, simple, sensitive, and accurate, which provides a basis for the investigation and monitoring of the pollution level of antibiotics in water. KEY WORDS: quinolone residues; antibiotics; water; high performance liquid chromatography mass spectrometry mass spectrometry 1 引言 (Quinolones) [1],,,,, ( ) [2],,,,, [3] (capillary electrophoresis, CE) [4-8] (high performance liquid chromatography, HPLC) - [9-17] (high performance liquid chromatography mass spectrometry mass spectrometry, HPLC-MS/MS), 14 - (HPLC-MS/MS) -,,, 14 -,,, 2 材料与方法 2.1 仪器 试剂与材料 UHPLC LC 30A ( SHIMADZU ); Q-trap 5500 - ( AB SCIEX ); ( Sigma ); ( Merck ); MS ( Fisher ) ( ) (fleroxacin) (lomefloxacin) (gatifloxacin) (danofloxacin), (enoxacin)sigma, (difloxacin) (pefloxacin) (ofloxacin) (sarafloxacin) (flumequine) (enrofloxacin) (marbofloxacin) (ciprofloxacin) Dr.Ehrenstorfer, (orbifloxacin) 2.2 标准溶液配制 (1.0 mg/ml): 0.01g, 10 ml, 1.0 mg/ml, 20 (1.0 μg/ml): 25 μl 25 ml,, 1.0 μg/ml, 4 : 5 10 50 100
4968 6 200 500 1000 ng/l 2.3 样品处理 15 min, 1.5 ml, 14000 rpm/min 5 min, 0.22 μm, 2.4 实验条件 : Atlantis dc-c18 (4.6 mm 150 mm, 5 µm, Waters); : A (0.05%), B - (4:6, V:V); : 20%B (0~3.0 min); 20%B 70%B (3.0~6.5 min); 70%B 90%B (6.5~9.0 min); 90%B 20%B (9.0~9.1 min); 20%B (9.1~11.0 min) 0.5 ml/min; 40 ; 20 μl : : (ESI); : ; : (MRM); : 550 ; Curtain Gas: 35 psi; Ion Source Gas1: 55 psi; Ion Source Gas2: 55 psi; : 5.5 kv; 1 表 1 14 种喹诺酮类药物残留的离子参数 Tbale 1 Ion parameters of the 14 quinolone residues (m/z) (m/z) (V) (V) 370.2 362.2 321.1 332.2 360.2 352.2 358.2 396.2 400.2 386.2 262.2 363.2 326.2* 22 70 269.2 32 318.2* 28 130 261.2 36 303.2 14 140 232.2* 48 288.2 23 100 245.2* 29 316.2* 24 60 245.2 37 308.2* 24 70 265.2 32 340.2 26 60 283.2* 30 352.2* 26 110 295.2 29 356.2 32 70 299.2* 46 342.2* 23 100 299.2 38 244.2 24 80 202.2* 42 320.2* 22 110 277.2 29
12, : - 14 4969 (m/z) (m/z) (V) (V) 续表 1 334.2 376.2 290.2* 25 100 233.2 33 332.2* 24 118 289.1 31 : * 3 结果与讨论 3.1 质谱条件的优化, 100 ng/ml 14 10 μl/min, (ESI + ) (ESI - Q1,, 14 ESI +,,, MRM DP CE,, 2.4 3.2 色谱条件的优化 - - - - [16] [M+H] +, ESI +,,, 0.05% 0.1% 0.5% -,, (6:4 5:5 4:6, V:V), 4:6,, 0.05% + - (4:6, V:V), 14,,, 1 3.3 目标化合物提取方法的选择, ng/l, [9,18] - μg/l,, ;,,, 15 min, 0.22 μm, (10 20 50 μl),, 20 μl,, 3.4 方法的线性范围 相关系数 检出限 定量限和精密度, (Y) (X, ng/l), 5 ng/l~1000 ng/l, 10 ng/l~1000 ng/l, 20 ng/l~1000 ng/l, 50 ng/l~1000 ng/l; r 0.98, 13 0.99; (S/N=3) (S/N=10), 14 20 ng/l, 50 ng/l; 20 ng/l 100 ng/l 500 ng/l (n=6), 83.9%~110.0%, 2.4%~15.4% 2 3.6 实际样品的测定 17 (6 6 5 ), 17 14
4970 食品安全质量检测学报 Fig. 1 图 1 14 种喹诺酮类药物残留的多反应监测色谱图 Multiple reaction monitoring (MRM) chromatography of the 14 quinolone residues 第6卷
第 12 期 吴 Fig. 1 健, 等: 高效液相色谱-质谱联用法同时检测水体中 14 种喹诺酮类药物残留 续图 1 14 种喹诺酮类药物残留的多反应监测色谱图 Multiple reaction monitoring (MRM) chromatography of the 14 quinolone residues 4971
4972 6 Table 2 表 2 14 种喹诺酮类药物残留的 LOD LOQ r 线性方程 回收率及 RSD LOD, LOQ, linear correlation coefficient(r), linear equation, recoveries and relative standard deviation(rsd) for 14 quinolone residues (ng/l) (ng/l) (r) /RSD(%, n=6) 20 ng/l 100 ng/l 500 ng/l 10 20 0.9992 Y = 610.1 X 3178.0 2 5 0.998 Y = 174.4X+210.12 10 20 0.994 Y = 221.7X+501.74 5 10 0.98 Y = 474.5 X 635.39 2 5 0.997 Y = 2217.9 X+227.62 2 5 0.997 Y = 3952.3 X+3465.8 20 50 0.996 Y = 154.8 X 535.44 5 10 0.998 Y = 608.0 X 1832.2 2 5 0.998 Y = 1340.2 X+61.776 2 5 0.996 Y = 945.2 X 1634.5 2 5 0.997 Y = 3508.9X+9297.3 5 10 0.990 Y = 733.2X+1110.5 5 10 0.994 Y = 1137.9 X 407.93 1 5 0.998 Y = 1584.9 X+2411.8 110.0 96.5 99.8 12.0 7.9 3.2 95.1 87.4 94.0 6.7 4.4 2.4 85.0 94.6 89.4 10.1 7.7 3.0 86.0 83.9 99.0 9.3 12.4 5.0 90.7 90.0 92.0 15.4 5.2 3.4 95.5 89.7 93.6 4.2 6.6 2.6 85.5 89.0 15.0 7.3 84.0 94.7 98.0 10.6 3.7 3.3 94.7 94.3 100.2 8.0 6.1 3.1 99.3 93.5 92.3 6.5 5.0 2.5 99.5 100.8 96.6 5.7 4.8 2.7 97.0 86.7 91.0 6.9 5.2 3.9 89.5 91.3 91.5 9.7 5.8 2.8 97.1 92.3 95.8 8.8 5.3 4.2
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