7 2 Vol. 7 No. 2 2016 2 Journal of Food Safety and Quality Feb., 2016 刘丽丽 *, 刘谦, 王宇 (, 071001) 摘 要 : 目的, 方法, HLB, C 18, ESI 结果 0.1~10 μg/kg, 0.999, 0.1 0.2 10.0 μg/kg 3, 95.0%~103.2%, 3.72%~6.85%, 0.1 μg/kg. 结论, 关键词 : ; ; Optimization of detection conditions of chloramphenicol in aquatic products LIU Li-Li *, LIU Qian, WANG Yu (Baoding Entry-Exit Inspection and Quarantine Bureau, Baoding 071001, China) ABSTRACT: Objective To establish a method for the determination of chloramphenicol in aquatic products by high performance liquid chromatography- tandem mass spectrometry, and to optimize the sample pretreatment such as extraction, cleanup and mobile phase. Methods Samples were extracted with ethyl acetate, cleaned up by the Oasis HLB SPE cartridges and collected by the C 18 chromatographic column. Finally, the chloramphenicol was detected in the ESI negative ion mode. Results The linear ranges were 0.1 10 μg/kg and the correlation coefficients were all above 0.99. The recoveries of chloramphenicol in aquatic products at the spiked levels of 0.1, 0.2 and 10 μg/kg were in ranges of 95.0% 103.2%. The relative standard deviations were 3.72%~6.85%, and the limit of detection was 0.1 μg/kg. Conclusion The method is reliable and stable, which can meet the requirements for the determination of chloramphenicol in aquatic products. KEY WORDS: high performance liquid chromatography-tandem mass spectrometry; chloramphenicol; processed aquatic products 1 引言, [1], [2] ;,,,,, [3] [4] [5] [6] * 通讯作者 :,, E-mail: 13931697750@163.com *Corresponding author: LIU Li-Li, Assistant Engineer, Baoding Entry-Exit Inspection and Quarantine Bureau, Baoding 071001, China. E-mail: 13931697750@163.com
624 7 [7-9],, ;,, ;, [10] ;,,,,, 2 材料与方法 2.1 材料 试剂与仪器 2.1.1 材料 ( ) 2.1.2 试剂 (, ); (, ); ( (100 μg/ml, SB05-093-2008 ), 10 μg/ml 20 ng/ml, (d5- ) 100 μg/ml( ), 10 μg/ml 1 μg/ml 2.1.3 仪器 Waters2695 WatersTQD (ESI)( Waters ); ( ); ( Sigma ); ( ); ( Agilent ); HLB(60mg,3ml) ( Waters ) 2.2 样品处理 2.2.1 样品制备, 2.2.2 提取 5 g( 0.01 g), 50 ml, 1 μg/ml 25 μl, 10 ml, 0.45 ml, 10 min, 4000 r/min 5 min, 25 ml ; 10 ml, 25 ml, 25 ml,, 5 ml, 45 2.2.3 净化 5 ml,, HLB ( ), HLB 5 ml, 10 ml,,, 10 ml,,, 5 ml, 10 ml, 45, 1 ml, 2.2.4 标准曲线的绘制 25 50 100 250 500 μl 20 ng/ml, 5 μl 1 μg/ml, 45, 1 ml, 2.3 仪器条件 2.3.1 液相条件 : C 18 (100 mm 2.1 mm, 5 μm); : : =33:67(V:V) ; : 35 ; : 30 μl 2.3.2 质谱条件 : (ESI-),, 3.00 kv; 35 V, : 120 ; : 450 ; : 500 L/h; : 50 L/h (m/z) 1 表 1 质谱参数 Table 1 Parameters of MS (m/z) (V) (V) 320.9/152* 33 18 320.9/257.1 33 11 326/157.2 33 8 * 3 结果与讨论 3.1 提取溶剂的选取,,
2, : 625,, ;,, 3.2 净化条件的优化 HLB,,,,,,, 60%,,,, HLB,,, 95%,, 3.3 流动相的优化 C 18, - -0.1% -10 mmol/l - -0.1% -10 mmol/l,,, ESI, - 3.4 质谱条件的优化 C 11 H 12 C 12 N 2 O 5, (ESI) H +, ESI, m/z 200~400, 320.9, 320.9,, 152 ( 1), 257.1 ( 2),, 3.5 线性关系 灵敏度和精密度, ( 2), 0.10~10 μg/kg, Y=1.24058X- 0.00868135, 0.999, 0.1 μg/kg, 3 4,,, 1 (0.1 ng/ml) ( ) Fig.1 Qualitative ion current of chloramphenicol (0.1 ng/ml)
626 7 2 (0.1ng/mL) ( ) Fig. 2 Quantitative ion current of chloramphenicol (0.1 ng/ml) Table 2 表 2 样品中氯霉素回收率及相对标准偏差 (n=6) Recoveries and RSD of chloramphenicol in samples (n=6) (μg/kg) (%) (%) 0.1 95.0 4.63 0.2 103.2 6.85 10.0 97.0 3.72 0.1 95.8 4.34 0.2 99.5 5.28 10.0 101.9 7.01 Fig. 3 3 (0.1 ng/ml) The total ion chromatogram of chloramphenicol ( 0.1 ng/ml)
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