7 1 Vol. 7 No. 1 2016 1 Journal of Food Safety and Quality Jan., 2016 肖家勇 1, 李莉 2, 丁利 1, 朱绍华 1, 龚强 1, 付善良 1, 王利兵 (1.,,, 410004; 2., 100176) 1* 摘要 : 目的 6 - (HPLC-MS/MS) 方法 Agilent ZORBAX SB-C 18, -0.1%,, 0.35 ml/min; (ESI+), (MRM);, 结果 6 (r 2 0.999), 0.04~0.2 μg/kg, 0.01~0.04 μg/kg 81.9%~103.9%, 7.2% 结论,, 6 关键词 : ; - ; Determination of phthalates in food by gel permeation chromatography-high performance liquid chromatography-tandem mass spectrometry XIAO Jia-Yong 1, LI Li 2, DING Li 1, ZHU Shao-Hua 1, GONG Qiang 1, FU Shan-Liang 1, WANG Li-Bing 1* (1. Technology Center of Hunan Entry-Exit Inspection and Quarantine Bureau, State Key Laboratory of Food Safety Testing, Hunan Key Laboratory of Food Safety Science & Technology, Changsha 410004, China; 2. Chinese Academy of Inspection and Quarantine, Beijing 100176 China) ABSTRACT: Objective To establish a method for the simultaneous determination of 6 kinds of phthalates in food by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). Methods The samples were separated by an Agilent ZORBAX SB-C 18 column with a mobile phase of methanol-0.1% formic acid/water in gradient elution mode at the flow rate of 0.35 ml/min. The mass spectrometry worked in the positive ion mode, and multiple reaction monitoring (MRM) was adopted for the quantification of the analytes. Samples were purified by gel permeation chromatography (GPC) after extraction with organic solvents. Results A good linearity for 6 kinds of phthalates was achieved, with all correlation coefficients more than 0.999. Average recoveries of 6 analytes at three spiked level were ranged from 81.9% to 103.9%, with RSDs less than 7.2%. The limits of quantification and detection of the method was 0.04~0.2 μg/kg and 0.01~0.04 μg/kg, respectively. Conclusion The established method is sensitive and simple, and suitable for 基金项目 : (2012BAD29B05) Fund: Supported by the National Key Technology Research and Development Program of the Ministry of Science and Technology of China (2012BAD29B05) * 通讯作者 :,, E-mail: wanglb0419@126.com *Corresponding author: WANG Li-Bing, Researcher, Hunan Key Laboratory of Food Safety Science Technology, Technology Center of Hunan Entry-Exit Inspection and Quarantine Bureau, Changsha 410004, China. E-mail: wanglb0419@126.com
92 7 the determination of 6 kinds of phthalates in food. KEY WORDS: phthalates; high performance liquid chromatography-tandem mass spectrometry; gel permeation chromatography 1 引言 (phthalic acid esters, PAEs),,, [1-3], PVC,,, 60%,,, [4,5] [6],, - [7-10],,, -, - [11] (DHXP)(BBP) (DCHP)(DHP) (DPRP) (DBEP) 6, - 6,, 2 材料与方法 2.1 材料 2.1.1 试验材料, 2.1.2 仪器与试剂 API 4000 ( ); 20A (); ( Sigma ); (Turbo Vap II, Caliper ); GPC (GPC Vario, Lctech ); (KQ-250E, ) HPLC ( Merck ), ( ); (DHXP) (BBP) (DCHP) (DHP) (DPRP) (DBEP) ( 99%, Sigma ); ( Millipore ) 2.2 方法 2.2.1 标准溶液的配制 (DHXP) (BBP) (DCHP) (DHP) (DPRP) (DBEP) 10.0 mg, 10 ml, 1.0 mg/ml, 4 1 ml 10 ml,, 100 μg/ml, 4, 1.0 μg/ml 2.2.2 仪器条件 : Agilent ZORBAX SB-C 18 (100 mm 2.1 mm, 3.5 μm); : A, B 0.1%, : 0~10.0 min, 50%~2%B, 10.0~15.0 min, 2%B, 15.1 min, 50%B, 15.0~20.0 min, 50%B; : 10 μl; : 0.35 ml/min; : 35 (ESI); : ; (Gas1): 413 kpa(60.00 psi); (Gas2): 448 kpa(65.00 psi); (IS): 4500.00
1, : -- 93 V; (Cur): 172 kpa(25.00 psi); (TEM): 500 ; : (MRM) 2.2.3 样品前处理 5.0 g 20 ml, 10 ml 2,, 40, 7.0 ml - (1:1, V:V), GPC GPC - (1:1, V:V), 5.0 ml/min; 840 960 300 s; 6400 μl GPC 40, 1.0 ml, 0.45 µm HPLC-MS/MS 3.2 质谱条件的优化 (ESI+), (FIA), 6 (MRM), 6, 1 6 MRM 2 表 1 多反应检测扫描模式检测 6 种邻苯二甲酸酯类物质的质谱参数 Table 1 MS parameters for the analysis of 6 kinds of PAEs m/z m/z DP/V CE/V EP/V CXP/V 3 结果与讨论 3.1 色谱条件的优化, 6, ESI+,,,, 6,, ( 1), -0.1% C 18,,,, 98%, DPRP 251.4 BBP 313.3 DCHP 331.2 DHXP 335.3 DHP 363.6 DBEP 367.4 * 149* 19 10 10 53 191.3 12 10 10 205.1* 11 10 8 149.1 70 18 10 7 239.1 10 10 10 149.1* 30 10 10 230.8 50 13 10 15 249.2 14 10 15 149.3* 19 10 10 70 233.1 12 10 10 149* 19 10 10 60 247.4 12 10 10 249.3* 11 10 10 75 101.1 17 10 10 1 Fig. 1 Effect of mobile phase modifier
94 7 3.4 干扰的排除,, ;, ; 24 h 2, 2 6 (a 5 ng/ml, b ) Fig. 2 MRM chromatograms of 6 kinds of PAEs (a: 5 ng/ml standard solution, b: blank solution) 3.5 线性范围与检测限 6, 2.2.2,,, 6, 0.999 (LOD) (LOQ) 3 10 (S/N), 2 2,, GB/T 21911-2008, 0.01~0.04 μg/kg 3.3 GPC 条件的优化 6 ( 0.1 µg/ml), 2.2.3 GPC, 12 min, 12 min 2 min, 2.2.2 HPLC-MS/MS, 6 ( 3) 3, 6 14~30 min 表 2 6 中邻苯二甲酸酯的线性范围 检测限及定量限结果 Table 2 Linear ranges, limit of detection and limit of quantification of 6 kinds of PAEs (ng/ml) LOD(μg/kg) LOQ(μg/kg) DPRP 0.2~100 0.01 0.04 BBP 0.2~100 0.01 0.04 DCHP 0.2~100 0.01 0.04 DHXP 0.2~100 0.01 0.04 DHP 0.2~100 0.01 0.04 DBEP 1.0~100 0.04 0.2 3 6 Fig. 3 Elution curve of 6 kinds of PAEs
1, : -- 95 3.6 准确度与精密度 6,, 4 24 h, 3,, 6, (RSD%) 3 3.7 实际样品的检测 ( 6 3 2 5 2 1 ) 6, 6 DPRP DHXP DBEP 19, BBP DCHP ( 42.1% 31.6%), 0.04~0.77 µg/kg, DHP ( 2 ), 0.04 µg/kg 4 4 结论 - (HPLC-MS/MS) 6,,, 表 3 3 个添加水平的回收率和精密度 (n=6) Table 3 Recoveries and precisions for blank sample spiked at 3 different levels (n=6) DPRP BBP DCHP DHXP DHP DBEP (µg/kg) (%) RSD(%) 2.0 81.9 3.0 5.4 20.0 85.3 2.4 7.8 50.0 90.2 5.9 8.3 2.0 93.6 4.4 6.7 20.0 91.7 3.4 5.5 50.0 102.5 4.9 4.9 1.0 103.9 5.7 7.3 20.0 99.3 3.6 5.7 50.0 98.2 6.9 7.1 1.0 89.6 5.5 6.1 20.0 93.7 7.2 7.9 50.0 91.5 4.4 6.7 1.0 87.0 5.6 6.5 20.0 92.3 6.4 7.7 50.0 95.9 6.9 9.6 1.0 91.7 1.9 6.7 20.0 89.3 3.2 5.5 50.0 93.5 3.0 4.6 4 MRM Fig. 4 MRM chromatogram of sample
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