Microsoft Word 欧阳少伦-二校

Transcription

1 6 5 Vol. 6 No Journal of Food Safety and Quality May., 2015 欧阳少伦 *, 邵琳智, 谢敏玲, 吴映璇, 林峰 (, ) 摘要 : 目的 (DMS) 方法,, PBA, 5 mmol/l ( 0.1% ) -, DMS ESI 结果 0.5~5.0 µg/l, 1.0 µg/l, µg/l, 68.4%~112.8%, (n=6) 7.3%~11.2% 结论,, 关键词 : ; ; Differential mobility spectrometry for analysis of amantadine and ribavirin residues in chicken muscle OUYANG Shao-Lun *, SHAO Lin-Zhi, XIE Min-Ling, WU Ying-Xuan, LIN Feng (Technical Center of Guangdong Entry-Exit Inspection and Quarantine Bureau, Guangzhou , China) ABSTRACT: Objective To establish a differential mobility spectrometry method for determination of amantadine and ribavirin residues in chicken muscle. Methods The amantadine and ribavirin in sample were extracted by trichloroacetic and enzyme hydrolyzing with acid phosphatase, then purified with PBA cartridge. The targeted compounds were separated on a HILIC column with the gradient elution of 5 mmol/l ammoniumacetate (0.1% formic acid) and acetonitrile as the mobile phases. Quantitative and qualitative were detected by tandem mass spectrometry in positive electrospray ionization and multiple reaction monitoring (MRM) mode with differential mobility spectrometry (DMS) on. Results The method showed a good linearity in the range of 0.5~5.0 µg/l, and the limits of quantification was 1.0 μg/kg. The recoveries in chicken muscle at the 1.0, 2.0 and 10 μg/kg spiked levels was in the range of 68.4%~112.8% with the relative standard deviations (RSD) in the range of 7.3%~11.2%. Conclusion This method can significantly improve the selection of target analysts, effectively eliminate the matrix interference and reduce the noise. KEY WORDS: differential mobility spectrometry; antiviral drugs; chicken muscle 基金项目 : (2014GDK01) Fund: Supported by the Scientific and Technological Project of Guangdong Entry-Exit Inspection and Quarantine Bureau (2014GDK01) * 通讯作者 :,, oyshl@163.com *Corresponding author: OUYANG Shao-Lun, Senior Engineer, Technical Center of Guangdong Entry-Exit Inspection and Quarantine Bureau, Guangzhou , China. oyshl@163.com

2 5, : 引言 -, LC-MS/MS (liquid chromatography-mass spectrometry),, LC-MS/MS [1,2],, [3-6],,,,,, [7-9], [5] (differential mobility spectrometry, DMS), LC-ESI-DMS-MS/MS (differential mobility spectrometry, DMS),,, Q0,,,, [10],,, 2 材料与方法 2.1 仪器与试剂 (UFLC LC-20AD, ); / (AB SCIEX QTRAP 5500, AB SCIEX ), (ESI) (AB SCIEX SelexION TM ); (MS3, IKA ); (T25, IKA ); (4K15, Sigma ) ; BOND ELUT-PBA (100 mg, 3 ml, ) -D 6-13 C 5 CHROMADEX Dr. Ehrenstorfer CDN TRC, 96%,, ( ), : 2.2 样品处理 5 g ( )( 0.01 g) 50 ml, -D 6-13 C 5, 15 min 20 g/l 10 ml, 1 min, 1 min, r/min 5 min,, 10 ml, 3 min, r/min 5 min, 20 ml 4 ml, ph 4.8, 2 ml ph mmol,, 25 μl 37 2 h, ph 8.5, 5 min, 3 ml 3 ml ph mmol PBA,, 3 ml ph mmol 3 ml,, 2 ml - (V:V=80:20, 0.1% ), 0.22 μm 2.3 色谱与质谱条件 : HILIC Silica (3.0 mm 100 mm, 3 μm); : 40 ; : 5 mmol/l ( 0.1% ) (A) (B) : 0 5 min, 10%A; 5 8 min, 10%A 50%A; min, 50%A 10%A; min, 10%A : 10 µl; : 300 µl/min : (ESI) ; : ; : (MRM); : 30 L/min; (GS1) : 75 L/min; (GS2) : 65 L/min; (CAD) : ; (TEM): 600 ; (IS): 5000

3 V; (MD): ; (MDC): 100 µl/min; (SV): 结果与讨论 3.1 未开启 DMS 情况下的测定 标准溶液的测定,, (HILIC) [11,12], ( 0.1% ),, MRM 鸡肉样品的测定,, MRM 2 A,,,,,, HILIC,,, 干扰物的分析 MRM,,,,,,,,, [13,14],, [6], 2 [15],, 3.2 采用 DMS 后的测定 DMS 质谱参数的优化 DMS, ESI MRM, DMS, 0.5 mg/l, SV, COV,,,, SV, COV, COV,,,,,, 鸡肉样品的测定 LC-ESI-DMS- MS/MS, MRM 2 B,,, DMS,,, (S/N), µg/kg (152.2/ /96.0) 表 1 金刚烷胺和利巴韦林及其内标物的质谱参数 Table 1 MS /MS parameters for the amantadine, ribavirin and internal standard (m/z) (m/z) (V) (V) (V) Amantadine Ribavirin * * *. Amantadine-D Ribavirin- 13 C

4 5, : 1709 Fig. 1 1 (0.5 μg/l) (MRM) Chromatograms of a mixture of amantadine and ribavirin standard solution (0.5 μg/l) in MRM mode Fig µg/kg MRM MRM chromatograms of spiked chicken musle 1.0 µg/kg with amantadine and ribavirin (A: - ; B: - ) (A: DMS-OFF; B: DMS-ON)

5 Fig µg/kg MRM MRM chromatograms of spiked chicken musle 1.0 µg/kg with amantadine and ribavirin (A: - ; B: - ) (A: DMS-OFF; B: DMS-ON)

6 5, : 方法的线性范围 定量限 加标回收率和精密度 (X), (Y) 0.5~5.0 µg/l, Y=0.158X (r=0.9996) Y=0.527X (r=0.9994), (LOQ), (S/N) 10, 3,, 1.0 µg/kg µg/kg, 6,, 2 表 2 空白鸡肉中金刚烷胺和利巴韦林的加标回收率与精密度 (n=6) Table 2 Recoveries and relative standard deviations (RSD) of amantadine and ribavirin spiked in blank chicken muscle sample (n=6) /(µg/kg) /% RSD/% 3.4 实际样品的测定 ~ ~ ~ ~ ~ ~ , µg/kg, MRM 3 Fig. 3 3 MRM MRM chromatograms of chicken muscle sample of a mixture of amantadine and ribavirin in MRM mode

7 结论,,,,,,,, 参考文献 [1] DB32/T [S]. DB32/T Determination of total residues of ribavirin and its metabolites in chicken liver-liquid chromatographytandem mass spectrometry method [S]. [2] DB32/T [S]. DB32/T Determination of amantadine residues in chicken liver-liquid chromatography-tandem mass spectrometry method [S]. [3],,,. - [J]., 2013, 31(8): Yun H, Cui FY, Yan H, et al. Determination of ribavirin and amantadine in chicken by ultra performance liquid chromatographytandem mass spectrometry [J]. Chin J Chromatogr, 2013, 31(8): [4],,,. - [J]., 2013, 31(10): Zhu WX, Yang JZ, Yuan P, et al. Rapid determination of total residues of ribavirinand its metabolites in chicken and chicken product by ultra-performance hydrophilic interaction chromatographytandem mass spectrometry [J]. Chin J Chromatogr, 2013, 31(10): [5],,,. - [J]., 2013, 32(12): Shao LZ, Yao YX, Xie ML, et al. Determination of amantadine and ribavirin in animal tissues by hydrophilic interaction chromatography-tandem mass spectrometry [J]. J Instrum Anal, 2013, 32(12): [6] Wei C, Grace JE Jr, Zvyaga TA, et al. Utility of high-resolution accurate MS to eliminate interferences in the bioanalysis of ribavirin and its phosphate metabolites [J]. Bioanal, 2012, 4(15): [7] Kromdijk W, Pereira SA, Rosing H, et al. Development and validation of an assay for the simultaneous determination of zidovudine, abacavir, emtricitabine, lamivudine, tenofovir and ribavirin in human plasma using liquid chromatography-tandem mass spectrometry [J]. J Chromatogr B, 2013, : [8] Zironi E, Gazzotti T, Lugoboni B, et al. Development of a rapid LC MS/MS method for ribavirin determination in rat brain [J]. J Pharm Bioned Anal, 2011, 54(4): [9] Meijuan X, Wenzheng J, Xiaoyan X, et al. Determination of rimantadine in rat plasma by liquidchromatography electrospray mass spectrometry and its application in a pharmacokinetic study [J]. J Chromatogr B, 2008, 864: [10]. AB SCIEX SelexION TM - [J]., 2011, 5: Li CB. AB SCIEX SelexION TM Technology [J]. Mod Sci Instrum, 2011, 5: [11],,,. - 5 [J]., 2012, 30(12): Liu ZC, Yang F, Yu KJ, et al. Multi-residue determination of five antiviral drugs in chicken tissues by liquid chromatographyelectrospray ionization tandem mass spectrometry [J]. Chin J Chromatogr, 2012, 30(12): [12],,,. /LC-MS/MS [J]., 2009, 15(6): Yun H, Zhang ZH, Luo SL, et al. Determination of amantadine residues in food of animal origin by Solid phase extraction /LC-MS/MS [J]. Mod Instrum, 2009, 15(6): [13] Yeh L T, Nguyen M, Dadgostari S, et al. LC MS/MS method for simultaneous determination of viramidine and ribavirin levels in monkey red blood cells [J]. J Pharm Biomed Anal, 2007, 43(3): [14] Meléndez M, Rosario O, Zayas B, et al. HPLC-MS/MS method for the intracellular determination of ribavirin monophosphate and ribavirin triphosphate in CEMss cells [J]. J Pharm Biomed Anal, 2009, 49(5): [15] Van der Lijke H, Alffenaar JWC, Kok WT, et al. Determination of ribavirin in human serum using liquid chromatography tandem mass spectrometry [J]. Talanta, 2012, 88(15): 作者简介 ( 责任编辑 : 李振飞 ) 欧阳少伦, 工程师, 主要研究方向为食品分析 oyshl@163.com