6 5 Vol. 6 No. 5 2015 5 Journal of Food Safety and Quality May., 2015 欧阳少伦 *, 邵琳智, 谢敏玲, 吴映璇, 林峰 (, 510623) 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, 1.0 2.0 10 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 510623, 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 :,, E-mail: oyshl@163.com *Corresponding author: OUYANG Shao-Lun, Senior Engineer, Technical Center of Guangdong Entry-Exit Inspection and Quarantine Bureau, Guangzhou 510623, China. E-mail: oyshl@163.com
5, : 1707 1 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, 10000 r/min 5 min,, 10 ml, 3 min, 10000 r/min 5 min, 20 ml 4 ml, ph 4.8, 2 ml ph 4.8 250 mmol,, 25 μl 37 2 h, ph 8.5, 5 min, 3 ml 3 ml ph 8.5 250 mmol PBA,, 3 ml ph 8.5 250 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; 8 8.1 min, 50%A 10%A; 8.1 13 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
1708 6 V; (MD): ; (MDC): 100 µl/min; (SV): 3250 1 3 结果与讨论 3.1 未开启 DMS 情况下的测定 3.1.1 标准溶液的测定,, (HILIC) [11,12], ( 0.1% ),, MRM 1 3.1.2 鸡肉样品的测定,, MRM 2 A,,,,,, HILIC,,, 3.1.3 干扰物的分析 MRM,,,,,,,,, [13,14],, [6], 2 [15],, 3.2 采用 DMS 后的测定 3.2.1 DMS 质谱参数的优化 DMS, ESI MRM, DMS, 0.5 mg/l, SV, COV,,,, SV, COV, COV,,,,,, 2.3 3.2.2 鸡肉样品的测定 LC-ESI-DMS- MS/MS, MRM 2 B,,, DMS,,, (S/N), 2 1.0 µg/kg (152.2/135.2 245.2/96.0) 42 4 115 382 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 152.2 Ribavirin 245.2 135.2 * 25 100 93.1 39 113.1 14 60 96.0 * 43-29 -13.5 *. Amantadine-D 6 158.2 141.2 100 25-29 Ribavirin- 13 C 5 250.2 113.1 60 14-13.5
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. 2 2 1.0 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)
1710 6 Fig. 2 2 1.0 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, : 1711 3.3 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-0.0369(r=0.9996) Y=0.527X-0.0286(r=0.9994), (LOQ), (S/N) 10, 3,, 1.0 µg/kg 1.0 2.0 10 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 1.0 68.4~93.6 10.4 2.0 71.9~107.2 7. 3 10.0 86.8~103.5 9.7 1.0 70.2~112.8 11.2 2.0 74.7~97.1 9.5 10.0 82.5~98.6 8.9 10, 1 3.8 µg/kg, MRM 3 Fig. 3 3 MRM MRM chromatograms of chicken muscle sample of a mixture of amantadine and ribavirin in MRM mode
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