高效液相色谱-串联质谱法快速测定动物源食品中的氯霉素残留量
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摘要
目的建立高效液相色谱-串联质谱联用法快速测定动物源食品中氯霉素残留量的分析方法。方法样品采用乙酸乙酯提取、空气吹干浓缩、正己烷脱脂后,经乙腈和水梯度洗脱、色谱柱Poroshell120EC-C_(18)(50 mm×2.1 mm, 2.7μm)分离、液相色谱-串联质谱仪测定,以氘代氯霉素为内标物进行定量。结果回归方程Y=0.177X+0.0008,相关系数r为0.9996;方法检出限和定量限分别为0.03μg/kg、0.1μg/kg;在0.1～1.0μg/kg浓度梯度范围内的平均回收率为91.4%～107.5%,相对标准偏差为4.5%～5.7%,2次能力验证结果满意。结论该方法经济、简便、快捷、稳定、准确性高,能满足大批量动物源性食品样品中氯霉素残留量测定的要求。
Objective To establish a method for the determination of chloramphenicol residues in animal derived food by high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS). Methods The animal derived food sample was extracted by ethyl acetate, the extraction liquid was enriched by air flow, and purified by n-hexane. The obtained target compound was separated by high performance liquid chromatography using Poroshell 120 EC-C_(18)(50 mm×2.1 mm, 2.7 μm) HPLC column with gradient elution of acetonitrile and water, and was detected by HPLC-MS/MS with multiple reaction monitoring mode(MRM). Internal standard method was used for quantitative analysis, and deuterated chloramphenicol was used for internal standard. Results The regression equation was Y=0.177 X+0.0008, and the correlation coefficient r was 0.9996. The limit of detection was0.03 μg/kg, and the limit of quantitation was 0.1 μg/kg. The recoveries for spiked standards in the concentration range of 0.1-1.0 μg/kg and the relative standard deviations were 91.4%-107.5% and 4.5%-5.7%, respectively. The results of two batches proficiency testing were satisfactory. Conclusion This method is economical, simple,rapid, effective and accurate, which is suitable for the determination of the content of chloramphenicol residue in animal derived food samples.
Objective To establish a method for the determination of chloramphenicol residues in animal derived food by high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS). Methods The animal derived food sample was extracted by ethyl acetate, the extraction liquid was enriched by air flow, and purified by n-hexane. The obtained target compound was separated by high performance liquid chromatography using Poroshell 120 EC-C_(18)(50 mm×2.1 mm, 2.7 μm) HPLC column with gradient elution of acetonitrile and water, and was detected by HPLC-MS/MS with multiple reaction monitoring mode(MRM). Internal standard method was used for quantitative analysis, and deuterated chloramphenicol was used for internal standard. Results The regression equation was Y=0.177 X+0.0008, and the correlation coefficient r was 0.9996. The limit of detection was0.03 μg/kg, and the limit of quantitation was 0.1 μg/kg. The recoveries for spiked standards in the concentration range of 0.1-1.0 μg/kg and the relative standard deviations were 91.4%-107.5% and 4.5%-5.7%, respectively. The results of two batches proficiency testing were satisfactory. Conclusion This method is economical, simple,rapid, effective and accurate, which is suitable for the determination of the content of chloramphenicol residue in animal derived food samples.
引文
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[2]中华人民共和国农业部.动物性食品中兽药最高残留限量[Z].农业部公告第235号,2002.The Ministry of Agriculture of the People's Republic of China.The maximum residue limit of veterinary drugs in animal food[Z].Announcement of the ministry of agriculture No.235,2002.
[3]中华人民共和国农业部.食品动物禁用的兽药及其它化合物清单[Z].农业部公告第193号,2002.The Ministry of Agriculture of the People's Republic of China.List of banned veterinary drugs and other compounds for food animals[Z].Announcement of the ministry of agriculture No.193,2002.
[4]彭运平,齐维,唐海波,等.应用酶联免疫法检测鱼肉、蜂蜜中氯霉素的残留量[J].现代食品科技,2010,26(12):1415-1418.Peng YP,Qi W,Tang HB,et al.Detection the residues of chloramphenicol in fish meat and honey using enzyme-linked immunosorbent assay[J].Mod Food Sci Technol,2010,26(12):1415-1418.
[5]谭慧,麦琦.酶联免疫分析法测定水产品中氯霉素残留量[J].中国卫生检验杂志,2010,20(7):1649-1650.Tan H,Mai Q.Determination of chloramphenicol residues in aquatic products by enzyme immunoassay assay[J].Chin J Health Lab Technol,2010,20(7):1649-1650.
[6]胡红美,郭远明,孙秀梅,等.超声波萃取-PSA净化-气相色谱法测定水产品中氯霉素[J].浙江海洋学院学报(自然科学版),2016,35(3):222-227.Hu HM,Guo YM,Sun XM,et al.Determination of chloramphenicol and florfenicol in fishery products by gas chromatography combined with ultrasonic extraction and PSA purification[J].J Zhejiang Ocean Univ(Nat Sci Ed),2016,35(3):222-227.
[7]吴明媛,杨姝丽,李小正,等.毛细管电子捕获气相色谱法同时测定水产品中氯霉素、甲砜霉素和氟甲枫霉素残留[J].广西农业科学,2009,40(5):567-570.Wu MY,Yang SL,Li XZ,et al.Simultaneous determination of residues of chloramphenicol,thiamphenicol and florfenicol in aquatic products by gas chromatographic method with electron capture detection[J].Agric Sci Guangxi,2009,40(5):567-570.
[8]邵会,冷凯良,周明莹,等.水产品中氯霉素、甲砜霉素、氟苯尼考、氟苯尼考胺多残留的同时测定-GC/MS法[J].渔业科学进展,2015,36(3):137-141.Shao H,Leng KL,Zhou MY,et al.Simultaneous determination of chloramphenicol,thiamphenicol,florfenicol and florfenicol amine residues in aquatic products with gas chromatography/mass spectrometry[J].Prog Fish Sci,2015,36(3):137-141.
[9]梁晓聪,王玮,田丽,等.气相色谱-串联质谱法测定淡水鱼中氯霉素残留量[J].中国卫生检验杂志,2015,25(7):940-942.Liang XC,Wang W,Tian L,et al.Determination of chloramphenicol residues in fresh water fish by GC-MS/MS[J].Chin J Health Lab Technol,2015,25(7):940-942.
[10]王安伟,覃锐,刘天密,等.高效液相色谱法检测罗非鱼中氯霉素的残留量[J].食品安全质量检测学报,2018,9(7):1665-1668.Wang AW,Qin R,Liu TM,et al.Determination of chloramphenicol residues in tilapia by high performance liquid chromatography[J].J Food Saf Qual,2018,9(7):1665-1668.
[11]占春瑞,郭平,陈振桂,等.超高效液相色谱法测定水产品中甲砜霉素和氟甲砜霉素残留[J].分析化学,2008,36(4):525-528.Zhan CR,Guo P,Chen ZG,et al.Determination of thiamphenicol and florfenico residues in aquatic products by ultra performance liquid chromatography[J].Chin J Anal Chem,2008,36(4):525-528.
[12]彭涛,李淑娟,储晓刚,等.高效液相色谱-串联质谱法同时测定虾中氯霉素、甲砜霉素和氟甲砜霉素残留量[J].分析化学,2005,33(4):463-466.Peng T,Li SJ,Chu XG,et al.Simultaneous determination of residues of chloramphenicol,thiamphenicol and florfenicol in shrimp by high performance liquid chromatography-tandem mss spectrometry[J].Chin JAnal Chem,2005,33(4):463-466.
[13]张凤清,王岩松,范世华,等.高效液相色谱-串联质谱法测定动物肌肉组织中氯霉素、甲砜霉素和氟苯尼考残留量[J].食品科学,2010,30(12):1329-1337.Zhang FQ,Wang YS,Fan SH,et al.High performance liquid chromatography-tandem mss spectrometric determination of chloramphenicol,thiamphenicol and florfenicol residues in animal muscles[J].Food Sci,2010,30(12):1329-1337.
[14]Vasiliy A,Anton K.A simple sample preparation for simultaneous determination of chloramphenicol and its succinate esters in food products using high-performance liquid chromatography/high-resolution mass spectrometry[J].Food Addit Contam A,2017,34(2):211-217.
[15]Tomasz?,Ma?gorzata G,Andrzej P.Analytical strategy for determination of chloramphenicol in different biological matrices by liquid chromatography-mass spectrometry[J].J Vet Res,2017,(61):321-327.
[16]柯庆青,李诗言,周凡,等.基于通过型固相萃取-高效液相色谱-串联质谱法同时测定水产品中的氯霉素和红霉素[J].食品科学,2017,38(24):241-246.Ke QQ,Li SY,Zhou F,et al.Simultaneous determination of chloramphenicol and erythromycin residues in aquatic products by high performance liquid chromatography-tandem mss spectrometry with oasis prime hlb solid phase extraction cleanup[J].Food Sci,2017,38(24):241-246.
[17]Posyniak A,Zmudzki J,Niedzielska J.Evaluation of sample preparation for control of chloramphenicol residues in porcine tissues by enzyme-linked immunosorbent assay and liquid chromatography[J].Anal Chim Acta,2003,483(1/2):307-311.
[18]沈美芳,赵文亚,费志良,等.用酶联免疫法测定水产品中氯霉素的残留量[J].南京师范大学学报:工程技术版,2003,3(2):1-4.Shen MF,Zhao WY,Fei ZL,et al.Enzyme linked immuno sorbent assay(ELISA)for quantitative analysis of chloramphenicol residues in aquatic products[J].J Nanjing Norm Univ(Eng Technol),2003,3(2):1-4.
[19]庄宛,叶玫.动物性食品中氯霉素(CAP)残留检测方法概述[J].福建畜牧兽医,2004,26(1):8-9.Zhuang W,Ye M.Overview of determination methods of chloramphenicol(CAP)residues in animal food[J].Fujian Anim Husb Vet Med,2004,26(1):8-9.
[20]GB/T 20756-2006可食动物肌肉、肝脏和水产品中氯霉素、甲砜霉素和氟苯尼考残留量的测定液相色谱-串联质谱法[S].GB/T 20756-2006 Method for the determination of chloramphenicol,thiamphenicol,and florfenicol residues in edible animal muscles,liver and aquatic products-LC-MS-MS method[S].
[2]中华人民共和国农业部.动物性食品中兽药最高残留限量[Z].农业部公告第235号,2002.The Ministry of Agriculture of the People's Republic of China.The maximum residue limit of veterinary drugs in animal food[Z].Announcement of the ministry of agriculture No.235,2002.
[3]中华人民共和国农业部.食品动物禁用的兽药及其它化合物清单[Z].农业部公告第193号,2002.The Ministry of Agriculture of the People's Republic of China.List of banned veterinary drugs and other compounds for food animals[Z].Announcement of the ministry of agriculture No.193,2002.
[4]彭运平,齐维,唐海波,等.应用酶联免疫法检测鱼肉、蜂蜜中氯霉素的残留量[J].现代食品科技,2010,26(12):1415-1418.Peng YP,Qi W,Tang HB,et al.Detection the residues of chloramphenicol in fish meat and honey using enzyme-linked immunosorbent assay[J].Mod Food Sci Technol,2010,26(12):1415-1418.
[5]谭慧,麦琦.酶联免疫分析法测定水产品中氯霉素残留量[J].中国卫生检验杂志,2010,20(7):1649-1650.Tan H,Mai Q.Determination of chloramphenicol residues in aquatic products by enzyme immunoassay assay[J].Chin J Health Lab Technol,2010,20(7):1649-1650.
[6]胡红美,郭远明,孙秀梅,等.超声波萃取-PSA净化-气相色谱法测定水产品中氯霉素[J].浙江海洋学院学报(自然科学版),2016,35(3):222-227.Hu HM,Guo YM,Sun XM,et al.Determination of chloramphenicol and florfenicol in fishery products by gas chromatography combined with ultrasonic extraction and PSA purification[J].J Zhejiang Ocean Univ(Nat Sci Ed),2016,35(3):222-227.
[7]吴明媛,杨姝丽,李小正,等.毛细管电子捕获气相色谱法同时测定水产品中氯霉素、甲砜霉素和氟甲枫霉素残留[J].广西农业科学,2009,40(5):567-570.Wu MY,Yang SL,Li XZ,et al.Simultaneous determination of residues of chloramphenicol,thiamphenicol and florfenicol in aquatic products by gas chromatographic method with electron capture detection[J].Agric Sci Guangxi,2009,40(5):567-570.
[8]邵会,冷凯良,周明莹,等.水产品中氯霉素、甲砜霉素、氟苯尼考、氟苯尼考胺多残留的同时测定-GC/MS法[J].渔业科学进展,2015,36(3):137-141.Shao H,Leng KL,Zhou MY,et al.Simultaneous determination of chloramphenicol,thiamphenicol,florfenicol and florfenicol amine residues in aquatic products with gas chromatography/mass spectrometry[J].Prog Fish Sci,2015,36(3):137-141.
[9]梁晓聪,王玮,田丽,等.气相色谱-串联质谱法测定淡水鱼中氯霉素残留量[J].中国卫生检验杂志,2015,25(7):940-942.Liang XC,Wang W,Tian L,et al.Determination of chloramphenicol residues in fresh water fish by GC-MS/MS[J].Chin J Health Lab Technol,2015,25(7):940-942.
[10]王安伟,覃锐,刘天密,等.高效液相色谱法检测罗非鱼中氯霉素的残留量[J].食品安全质量检测学报,2018,9(7):1665-1668.Wang AW,Qin R,Liu TM,et al.Determination of chloramphenicol residues in tilapia by high performance liquid chromatography[J].J Food Saf Qual,2018,9(7):1665-1668.
[11]占春瑞,郭平,陈振桂,等.超高效液相色谱法测定水产品中甲砜霉素和氟甲砜霉素残留[J].分析化学,2008,36(4):525-528.Zhan CR,Guo P,Chen ZG,et al.Determination of thiamphenicol and florfenico residues in aquatic products by ultra performance liquid chromatography[J].Chin J Anal Chem,2008,36(4):525-528.
[12]彭涛,李淑娟,储晓刚,等.高效液相色谱-串联质谱法同时测定虾中氯霉素、甲砜霉素和氟甲砜霉素残留量[J].分析化学,2005,33(4):463-466.Peng T,Li SJ,Chu XG,et al.Simultaneous determination of residues of chloramphenicol,thiamphenicol and florfenicol in shrimp by high performance liquid chromatography-tandem mss spectrometry[J].Chin JAnal Chem,2005,33(4):463-466.
[13]张凤清,王岩松,范世华,等.高效液相色谱-串联质谱法测定动物肌肉组织中氯霉素、甲砜霉素和氟苯尼考残留量[J].食品科学,2010,30(12):1329-1337.Zhang FQ,Wang YS,Fan SH,et al.High performance liquid chromatography-tandem mss spectrometric determination of chloramphenicol,thiamphenicol and florfenicol residues in animal muscles[J].Food Sci,2010,30(12):1329-1337.
[14]Vasiliy A,Anton K.A simple sample preparation for simultaneous determination of chloramphenicol and its succinate esters in food products using high-performance liquid chromatography/high-resolution mass spectrometry[J].Food Addit Contam A,2017,34(2):211-217.
[15]Tomasz?,Ma?gorzata G,Andrzej P.Analytical strategy for determination of chloramphenicol in different biological matrices by liquid chromatography-mass spectrometry[J].J Vet Res,2017,(61):321-327.
[16]柯庆青,李诗言,周凡,等.基于通过型固相萃取-高效液相色谱-串联质谱法同时测定水产品中的氯霉素和红霉素[J].食品科学,2017,38(24):241-246.Ke QQ,Li SY,Zhou F,et al.Simultaneous determination of chloramphenicol and erythromycin residues in aquatic products by high performance liquid chromatography-tandem mss spectrometry with oasis prime hlb solid phase extraction cleanup[J].Food Sci,2017,38(24):241-246.
[17]Posyniak A,Zmudzki J,Niedzielska J.Evaluation of sample preparation for control of chloramphenicol residues in porcine tissues by enzyme-linked immunosorbent assay and liquid chromatography[J].Anal Chim Acta,2003,483(1/2):307-311.
[18]沈美芳,赵文亚,费志良,等.用酶联免疫法测定水产品中氯霉素的残留量[J].南京师范大学学报:工程技术版,2003,3(2):1-4.Shen MF,Zhao WY,Fei ZL,et al.Enzyme linked immuno sorbent assay(ELISA)for quantitative analysis of chloramphenicol residues in aquatic products[J].J Nanjing Norm Univ(Eng Technol),2003,3(2):1-4.
[19]庄宛,叶玫.动物性食品中氯霉素(CAP)残留检测方法概述[J].福建畜牧兽医,2004,26(1):8-9.Zhuang W,Ye M.Overview of determination methods of chloramphenicol(CAP)residues in animal food[J].Fujian Anim Husb Vet Med,2004,26(1):8-9.
[20]GB/T 20756-2006可食动物肌肉、肝脏和水产品中氯霉素、甲砜霉素和氟苯尼考残留量的测定液相色谱-串联质谱法[S].GB/T 20756-2006 Method for the determination of chloramphenicol,thiamphenicol,and florfenicol residues in edible animal muscles,liver and aquatic products-LC-MS-MS method[S].