固相萃取高效液相色谱串联质谱法同时测定蜂蜜中甲硝唑、氯霉素、甲砜霉素和氟甲砜霉素残留

Simultaneous determination of of metronidazole,chloramphenicol,thiamphenicol and florfenicol residues in honey by solid-phase extraction coupled with high performance liquid chromatography-tandem mass spectrometry

目的建立蜂蜜中甲硝唑(MTZ)、氯霉素(CAP)、甲砜霉素(TAP)和氟甲砜霉素(FF)残留的固相萃取液相色谱质谱分析方法。方法以氘代氯霉素(D5-CAP)和氘代甲硝唑(D4-MTZ)为内标,试样经乙酸乙酯提取,MCS固相萃取柱净化,用乙酸乙酯洗脱氯霉素类抗生素,5%氨化甲醇洗脱甲硝唑,吹干浓缩后,分别先加入0.10 ml甲醇超声溶解残留物,再加入0.90 ml 10%甲醇/水溶液混匀,经Shim-pack XR-ODS C18色谱柱分离,以0.1%甲酸水-甲醇做流动相,梯度洗脱,氯霉素类抗生素采用ESI-监测模式、甲硝唑采用ESI+监测模式,均采用同位素内标法定量。结果该方法在0.1μg/L^10μg/L呈良好的线性,相关系数均>0.999,甲硝唑、氯霉素、甲砜霉素和氟甲砜霉素的回收率为90.6%~110.3%,相对标准偏差(RSD)为1.5%~6.1%,检出限为0.005μg/kg^0.02μg/kg,定量限为0.015μg/kg^0.06μg/kg。结论该方法操作简便、准确、检出限低、重现性好,适用于蜂蜜中甲硝唑、氯霉素、甲砜霉素和氟甲砜霉素残留的检测。

Objective To develop a method for the determination of residues of metronidazole（ MTZ）,chloramphenicol（ CAP）,thiamphenicol（ TAP） and florfenicol（ FF） in honey by solid phase extraction liquid chromatography mass spectrometry.Methods Coupled with internal standards of D5-chloramphenicol（ D5-CAP） and D4-metronidazole（ D4-MTZ）,the target antibiotics in samples were extracted with ethyl acetate,cleaned-up with MCS solid phase column. Chloramphenicol antibiotics were eluted with ethyl acetate,metronidazole was eluted with 5% ammoniated methanol. After dried,concentrated,and then 0. 10 ml methanol was added to the ultrasound to dissolve the residue respectively. Rejoined by 0. 90 ml 10% methanol/water mixture,chloramphenicol antibiotics and metronidazole were separated by gradient elution with 0. 1% formic acid water-methanol in Shim-pack XR-ODS C18 column. Chloramphenicol antibiotics and metronidazole were quantified by isotope internal standard method respectively in ESI-mode and ESI＋mode. Isometric internal standard method was used for quantification.Results The method showed good linearity in the range of 0. 1 μg/L-10 μg/L,with correlation coefficients（ r） exceeding0. 999. The recoveries for MTZ,CAP,TAP,FF were within 90. 6%-110. 3%,the relative standard deviations（ RSD） were within 1. 5%-6. 1%. The detection limits were within 0. 005 μg/kg-0. 02 μg/kg,and the limits of quantification were within0. 015 μg/kg-0. 06 μg/kg. Conclusion The method is simple,accurate,sensitive and producible,and it is suitable for the determination of metronidazole,chloramphenicol,thiamphenicol and florfenicol in honey.