Microsoft Word 甘蓓-二校-出3677

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1 6 9 Vol. 6 No Journal of Food Safety and Quality Sep., 甘蓓 1*, 况慧娟 2, 涂瑾 3 (1., ; 2., ; 3., ) 摘要 : 目的, - - 方法, 4, - 4 结果 %~83.5%, (RSD, n=5) 2.2%~16.0%, 0.13~0.79 ng/ml, 0.44~2.63 ng/ml, 1.0~25.0 µg/l ~ 结论, 关键词 : ; ; - ; ; Magnetic solid phase extraction-liquid chromatography-tandem mass spectrometry for the determination of 4 kinds of benzodiazepines residues in beef GAN Bei 1*, KUANG Hui-Juan 2, TU Jin 3 (1. Jiangxi Provincial Product Quality Supervision Testing College, Nanchang , China; 2. State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang , China; 3. College of Food Science and Engineering, Jiangxi Agricultural University, Nanchang , China) ABSTRACT: Objective To establish a novel method for the determination of 4 kinds of benzodiazepines (BZDs, chlordiazepoxide, diazepam, oxazepam, and triazolam) by magnetic solid phase extraction (MSPE)-liquid chromatography-tandem mass spectrometry (LC-MS/MS), based on the sulfo-functionalized magnetic beads. Methods After optimization of the extraction/desorption conditions, 4 BZDs were extracted with sulfofunctionalized magnetic beads, and detected by liquid chromatography-tandem mass spectrometry in multiple-reaction monitoring mode. Results The average recoveries of 4 benzodiazepines were 49.6%~83.5%, the relative standard deviation (RSD) of this method was between 2.2%~16.0%. The limits of detection (LOD) and limits of quantification (LOQ) of 4 kinds of benzodiazepines were in the range of 0.13~0.79 and 0.44~2.63 ng/ml, respectively. This method had a good linearity in the range of 1.0~25.0 μg/l between the peak areas and 基金项目 : (GZJK201202) Fund: Supported by Major Scientific and Technological Projects of the Jiangxi Provincial Bureau of Quality and Technical Supervision (GZJK201202) * 通讯作者 :,, ganbeikm@126.com *Corresponding author: GAN Bei, Engineer, Jiangxi Provincial Product Quality Supervision Testing College, Nanchang , China. ganbeikm@126.com

2 concentration in 4 kinds of benzodiazepines with the correlation coefficients were ~ Conclusion The method was fast, accurate and sensitive, which could be applied in the routine assay on benzodiazepines in beef. KEY WORDS: sulfo-functionalized magnetic beads; magnetic solid phase extraction; liquid chromatographytandem mass spectrometry; benzodiazepines; beef 1 引言 (benzodiazepines, BZDs) 1,4-,, [1, 2],, [3],, [4],,,, [3],, [5] (LC-MS/MS) [6-8] (LC-PDA) [9] (GG-MS) [10] (UPLC-PDA) [2],,,,, (LLE) [11,12] (SPE) [13,14] (SPME) [15] (MAE) [9], SPE, [16,17], (MSPE),,,,,,, LC-MS/MS, 4, 2 材料与方法 2.1 材料与试剂 4 98%,, (sulfo-mnbs), 10 mg(fe)/ml 2.2 仪器与设备 Agilent 6400 ( ); ( ); (JA2603B, ); (1-14, Sigma ); (Milli-Q, Milipore ) 2.3 实验方法标准工作储备液的配置 4 10 mg, g, 10 ml, 100 ml,, mg/l 10 mg, 10 ml, 0.1 mol/l HCl 100 mg/l mol/l HCl, 样品预处理 100 mg/l 100 µl 1.5 ml, 0.1 mol/l HCl 2 ng/ml

3 9, : ml,,, 5 min,, 1 ml, 0.22 µm 液相色谱和质谱条件 (1) : ZORBAX Eclipse XDB C 18 (4.6 mm 150 mm, 5 μm); : 40 ; : 0.3 ml/min; : 30 µl; : (A) 0.1% (B); 1 4,, 1 1, 2~25 µl, 4 ; 25 µl,, 25~40 µl, 25 µl 10 mg(fe)/ml 2 ng/ml 4 Table 1 表 1 流动相梯度洗脱程序 Program of gradient elution of mobile phase /min A/% B/% (2) : (ESI); : ; : (MRM); : 350 ; : 30 L/min; : 4000 V; 2 3 结果与分析 3.1 样品前处理的优化不同磁珠用量对回收率的影响 2 ng/ml µl 10 mg (Fe)/mL, 1 4 Fig. 1 Effect of amount of sulfo-mnbs on recoveries of 4 kinds of BZDs ph 值对回收率的影响, ph 1 mol/l NaOH HCl ( 2 ng/ml) ph , 25 µl,,, 2 表 2 4 种苯二氮卓类物质的离子对碰撞电压保留时间 Table 2 Ion pair, collision voltage, and retention time of 4 kinds of BZDs (m/z) (V) (min) 285.1/222.1, 285.1/193.1* 165, /241.1, 287.1/104.1* 135, /308.1, 343.1/239* 160, /241, 300.1/227* 110,

4 ph 4 Fig. 2 Effect of ph value on recoveries of 4 kinds of BZDs 4 3 3, 3 % ph, 3.2 方法学验证线性方程检出限定量限 μg /L, 4 1.0~25.0 μg/l, 4 (r 2 ) 0.999, (S/N 3) 0.13~0.79 ng/ml, (S/N 10) 0.44~2.63 ng/ml, 3 2, ph 1~6, 4 ph, ph=6.0,,,,,, [18],, 不同的洗脱液对回收率的影响 ph=6.0, 25 µl, - (99:1, 98:3, 97:4, V:V) - (97:3, V:V)4 a-1% ; b-3% ; c-4% ; d-3% 3 4 Fig. 3 Effect of different eluent on recoveries of 4 kinds of BZDs Table 3 表 3 4 种苯二氮卓类物质的线性方程相关系数检出限定量限 Linear regression equation, correlation coefficients, LOD and LOQ of 4 kinds of BZDs (r 2 ) (ng/ml) (ng/ml) Y= X Y= X Y= X Y= X

5 9, : 回收率和精密度, µg/kg 4, 5,, %~83.5%, 4 (RSD) 2.2%~16.0%, 4, 1 µg/kg 4 4 表 4 4 种苯二氮卓类物质的添加浓度平均回收率和相对标准偏差 (n=5) Table 4 Spiked level, average recovery and RSD for 4 kinds of BZDs (n=5) 1.0 µg/kg 2.0 µg/kg 5.0 µg/kg /% RSD/% /% RSD/% /% RSD/% ( 1.0 µg/kg) Fig. 4 MRM chromatograms of a blank beef spiked with the 4 kinds of BZDs at 1.0 µg/kg

6 方法的应用,,, 4 4 结论 - - 4,, -,, 参考文献 [1] Mahjoub AE, Staub C. Simultaneous determination of benzodiazepines in whole blood or serum by HPLC/DAD with a semi-micro column [J]. J Pharm Biom, 2000, 23 (2) : [2] Purificación F, Cristina GM, Teresa P, et al. A rapid ultrasound-assisted dispersive liquid liquid microextraction followed by ultra-performance liquid chromatography for the simultaneous determination of seven benzodiazepines in human plasma samples [J]. Anal Chim Acta, 2013, 767: [3],,,. [J]., 2007, 28(8): Li QS, Xu CL, Peng CF, et al. Progress on determination of benzodiazepines in animal product foods [J]. Food Sci, 2007, 28(8): [4],,, [J]., 2008, 27(11): Deng XJ, Guo DH, Zhu J, et al. Determination of benzodiazepines and metabolites residues in foodstuffs of animal origin by liquid chromatography-tandem mass spectrometry [J]. J Instrum Anal, 2008, 27(11): [5] Asthana A, Bose D, Durgbanshi A, et al. Determination of aromatic amines in water samples by capillary electrophoresis with electrochemical and fluorescence detection [J]. J Chromatogr A, 2000, 895(1 2): [6] Laurito TL, Mendes GD, Santagada V, et al. Bromazepam determination in human plasma by high-performance liquid chromatography coupled to tandem mass spectrometry: a highly sensitive and specific tool for bioequivalence studies [J]. J Mass Spectrom, 2004, 39(2): [7] Smink BE, Brandsma JE, Dijkhuizen A, et al. Quantitative analysis of 33 benzodiazepines, metabolites and benzodiazepine-like substances in whole blood by liquid chromatography (tandem) mass spectrometry [J]. J Chromatogr B, 2004, 811(1): [8] Bugey A, Staub C. Application of monolithic supports to online extraction and LC-MS analysis of benzodiazepines in whole blood samples [J]. J Sep Sci, 2007, 30(17): [9] Fernández P, Vázquez C, Lorenzo RA, et al. Experimental design for optimization of microwave-assisted extraction of benzodiazepines in human plasma [J]. Anal Bioanal Chem, 2010, 397(2): [10] Papoutsis II, Athanaselis SA, Nikolaou PD, et al. Development and validation of an EI GC MS method for the determination of benzodiazepine drugs and their metabolites in blood: Applications in clinical and forensic toxicology [J]. J Pharma Biom, 2010, 52(4): [11] Bugey A, Staub C. Rapid analysis of benzodiazepines in whole blood by high-performance liquid chromatography: use of a monolithic column [J]. J Pharm Biom, 2004, 35(3): [12] Ming DS, Heathcote J. A rapid and accurate UPLC/MS/MS method for the determination of benzodiazepines in human urine [J]. J Chromatogr B, 2011, 879(5): [13] Uddin MN, Samanidou VF, Papadoyannis IN. Development and validation of an HPLC method for the determination of six 1, 4-benzodiazepines in pharmaceuticals and human biological fluids [J]. J Liq Chromatogr RT, 2008, 31(9): [14] Fernandez P, Vázquez C, Lorenzo RA, et al. Development of a liquid chromatographic method for the simultaneous determination of six benzodiazepines in human plasma after solid-phase extraction [J]. Anal Lett, 2010, 43(6): [15] Luo Y, Pan L, Pawliszyn J. Determination of five benzodiazepines in aqueous solution and biological fluids using solid-phase microextraction with carbowaxtm/dvb fiber coating [J]. J Microcolumn Sep, 1998, 10(2): [16] Fernández P, Regenjo M, Fernández AM, et al. Optimization of ultrasound-assisted dispersive liquid liquid microextraction for ultra performance liquid chromatography determination of benzodiazepines in urine and hospital wastewater [J]. Anal Method, 2014, 6 (20):

7 9, : [17] Guo L, Zhang JW, Wei H, et al. Nanobeads-based rapid magnetic solid phase extraction of trace amounts of leuco-malachite green in Chinese major carps [J]. Talanta, 2012, 97: [18],,,. [J]., 2002, (4): Liu KL, Zhang CS, Zhou SG, et al. Summary of analysis methods of benzodiazepines in biological samples [J]. Foren Sci Technol, 2002, (4): 作者简介 ( 责任编辑 : 李振飞 ) 甘蓓, 工程师, 主要研究方向为食品安全检测技术 ganbeikm@126.com

g ml 10% ph 色谱条件 kinetex C μm 100 A 4. 6 mm 150 mm 25 5 μl A B 10 mmol /L ml /min

g ml 10% ph 色谱条件 kinetex C μm 100 A 4. 6 mm 150 mm 25 5 μl A B 10 mmol /L ml /min 25 4 2013 8 监测技术 5 1 2 1 ( 1. 江苏省疾病预防控制中心, 江苏南京 210009; 2. 南京医科大学第二附属医院, 江苏南京 210029) 5 Phenomenex kinetex C 18 2. 6 μm 4. 6 mm 150 mm 10 mmol /L ph 2. 5 + 60 40 DAD 280 nm 5 20. 0 mg /L ~ 100 mg /L 0.

Microsoft Word 甘 蓓-二校-出3677

Microsoft Word 甘蓓-二校-出3677