空间位阻净化-超高效液相色谱-串联质谱技术测定动物源性食品中抗组胺类药物残留
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摘要
建立超高效液相色谱-串联质谱法快速测定动物源性食品中19种抗组胺类药物及其代谢物残留的方法。样品用乙腈-乙酸乙酯溶液提取,提取液经空间位阻净化吸附剂EMR净化,再经无水硫酸镁和氯化钠盐析并浓缩后测定,以0.1%甲酸和0.1%甲酸-乙腈作为流动相进行梯度洗脱,在Poroshell 120 EC-C18色谱柱(100 mm×2.1 mm,2.7μm)上分离,柱温35℃,流速0.3 mL/min,进样量10μL,在电喷雾正离子模式下以动态多反应监测采集数据,外标法定量。结果表明,19种抗组胺类药物及其代谢物在相应质量浓度范围内线性良好,相关系数均大于0.999,不同阴性样品基质中(牛肉、鸡肉、猪肝)在3个质量浓度水平加标实验回收率在75.1%～89.1%之间,相对标准偏差为2.3%～7.1%,方法检出限为0.2～2.0μg/kg,定量限为0.6～6.0μg/kg。经方法学验证,本方法简便快速、重复性好、灵敏度高、结果可靠。适用于动物源性食品中19种抗组胺类药物及其代谢物的快速筛查和定量测定。
A rapid method for the determination of the residues of 19 anti-histamines and their metabolites in animal-derived foods by ultra performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS) was established. Samples were extracted with acetonitrile-ethyl acetate mixture, and the extract was purified with Enhanced Matrix Removal-Lipid sorbent(EMR-Lipid), and then salted out and concentrated with MgSO4 and NaCl. The chromatographic separation was accomplished on a Poroshell 120 EC-C18 column by gradient elution using 0.1% formic acid as mobile phase A and 0.1%formic acid-acetonitrile as mobile phase B at a flow rate of 0.3 mL/min. The column temperature was set at 35 ℃, and the injection volume was 10 μL. Data were collected by dynamic multi-reaction monitoring(MRM) in electrospray positive ion mode. The analytes were quantitated by external standard method. Statistical analysis indicated that the calibration curves for all analytes had a good linearity with correlation coefficients R2 greater than 0.999 in the tested concentration ranges. The average recoveries for different negative samples(beef, chicken and porcine liver) at three spiked levels were in the range of75.1%–89.1%, with relative standard deviations(RSDs) of 2.3%–7.1%. The limits of detection(LODs) of this method were0.2–2.0 μg/kg and the limits of quantitation(LOQs) were 0.6–6.0 μg/kg. In a word, this method proved to be simple, rapid,highly repeatable and sensitive, and reliable, and it could be suitable for rapid screening and quantitative determination of 19 antihistamines and their metabolites in anima-derived food samples.
A rapid method for the determination of the residues of 19 anti-histamines and their metabolites in animal-derived foods by ultra performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS) was established. Samples were extracted with acetonitrile-ethyl acetate mixture, and the extract was purified with Enhanced Matrix Removal-Lipid sorbent(EMR-Lipid), and then salted out and concentrated with MgSO4 and NaCl. The chromatographic separation was accomplished on a Poroshell 120 EC-C18 column by gradient elution using 0.1% formic acid as mobile phase A and 0.1%formic acid-acetonitrile as mobile phase B at a flow rate of 0.3 mL/min. The column temperature was set at 35 ℃, and the injection volume was 10 μL. Data were collected by dynamic multi-reaction monitoring(MRM) in electrospray positive ion mode. The analytes were quantitated by external standard method. Statistical analysis indicated that the calibration curves for all analytes had a good linearity with correlation coefficients R2 greater than 0.999 in the tested concentration ranges. The average recoveries for different negative samples(beef, chicken and porcine liver) at three spiked levels were in the range of75.1%–89.1%, with relative standard deviations(RSDs) of 2.3%–7.1%. The limits of detection(LODs) of this method were0.2–2.0 μg/kg and the limits of quantitation(LOQs) were 0.6–6.0 μg/kg. In a word, this method proved to be simple, rapid,highly repeatable and sensitive, and reliable, and it could be suitable for rapid screening and quantitative determination of 19 antihistamines and their metabolites in anima-derived food samples.
引文
[1]周瑾.我国兽药产业发展问题研究[D].保定:河北农业大学,2013.
[2]田鹏飞.国内外兽药企业创新与发展研究[D].武汉:华中农业大学,2012.
[3]闫小峰.浅谈我国兽药行业存在的主要问题及对策[J].中国兽药杂志,2009,43(8):35-38.DOI:10.3969/j.issn.1002-1280.2009.08.010.
[4]潘明飞,王俊平,方国臻,等.食品中农兽药残留检测新技术研究进展[J].食品科学,2014,35(15):277-282.DOI:10.7506/spkx1002-6630-201415056.
[5]车文军,张征,徐春祥,等.固相萃取/高效液相色谱法测定化妆品中5种抗组胺药物残留[J].分析测试学报,2012,31(8):1005-1008.DOI:10.3969/j.issn.1004-4957.2012.08.022.
[6]叶林虎,徐松,王丽莎,等.LC-MS/MS法检测皮肤科常用中药制剂中添加的7种抗组胺药物[J].药物分析杂志,2016,36(11):20-22.
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[8]林芳,王一欣,李涛,等.安神类保健食品中新型非法添加药物的检测研究[J].食品安全质量检测学报,2016,7(4):1631-1636.
[9]邱畅,温预关,李恒,等.LC-ESI-MS/MS法测定人血浆中西替利嗪浓度及其咀嚼片人体生物等效性评价[J].中国临床药学杂志,2008,17(4):234-238.DOI:10.3969/j.issn.1007-4406.2008.04.012.
[10]王伟,陈振玲,崔玉静,等.超高效液相色谱质谱法同时测定人血浆中8种抗组胺类药物[J].中国临床药理学杂志,2017,33(10):929-932.DOI:10.13699/j.cnki.1001-6821.2017.10.017.
[11]卫生部,中国标准化管理委员会.饲料中盐酸异丙嗪、盐酸氯丙嗪、地西洋、硫酸硫利达嗪、奋乃静的同步测定高效液相色谱法和液相色谱质谱联用法:NY/T 1458-2007[S].北京:中国标准出版社,2007.
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[13]BROEDERS J,BLAAUBOER B,HERMENS J.Development of a negligible depletion-solid phase microextraetion method to determine the free concentration of chlorpromazine inaqueous samples containing albumin[J].Chromatography A,2011,1218:8529-8535.
[14]李婧妍,郭春锋,崔立辉,等.QuEChERs-HPLC-MS/MS法检测液态乳中9种β2-受体激动剂残留量[J].中国乳品工业,2016,44(11):49-52.DOI:10.3969/j.issn.1001-2230.2016.11.013.
[15]李琴,陶大利,周红,等.超高效液相-串联质谱法同时测定生鲜乳中45种兽药残留量[J].中国乳品工业,2016,44(1):44-47.DOI:10.3969/j.issn.1001-2230.2016.01.010.
[16]路平,曲志娜,谭维泉,等.气相色谱-质谱法测定猪肝中氯丙嗪残留的研究[J].中国动物检疫,2006,23(7):30-11.DOI:10.3969/j.issn.1005-944X.2006.07.016.
[17]吕燕,杨挺,赵健,等.气相色谱-质谱法测定猪肝中氯丙嗪残留量[J].分析试验室,2008,27(3):119-121.DOI:10.3969/j.issn.1000-0720.2008.03.033.
[18]WEJDAN S K,BAHRUDDIN S,NORMALIZA H A M,et al.Microsolid phase extraction with liquid chromatograph-tandem mass spectrometry for the determination of aflatoxins in coffee and malt beverage[J].Food Chemistry,2014,147(4):287-294.DOI:10.1016/j.foodchem.2013.09.049.
[19]黄德凤,VAN TRAN K,MICHEAL S,等.Oasis PRIME HLB-UPLC-MS/MS测定猪肉中78种兽药残留[G]//Oasis PRIME HLB食品应用文集.2015:8-10.
[20]LI M,KONG W,LI Y,et al.High-throughput determination of multimycotoxins in Chinese yamand related products by ultra fast liquid chromatography coupled with tandem mass spectrometry after onestep extraction[J].Journal of Chromatography B,2016,1022(6):118-125.DOI:10.1016/j.jchromb.2016.04.014.
[21]孙伟华,杨欢,曹赵云,等.基于分散固相萃取液相色谱-串联质谱法测定大米中8种真菌毒素[J].分析测试学报,2017,36(1):47-53.DOI:10.3969/j.issn.1004-4957.2017.01.008.
[22]李诗言,柯庆青,王鼎南,等.QuEChERS-LC-MS/MS法测定水产配合饲料中4种常见黄曲霉毒素残留[J].中国渔业质量与标准,2017,7(5):25-31.DOI:10.3696/j.issn.2095-1833.2017.05.004.
[23]谢寒冰,蒋万枫,赵海峰,等.高效液相色谱法-四极杆飞行时间质谱法测定猪肉中20种兽药残留量[J].理化检验:化学分册,2014,50(6):702-707.
[24]梁玉禧,李益珍,郭小敏,等.液质联用法测定猪肉产品中违禁添加物氯丙嗪、异氯丙嗪和万古霉素残留研究[J].粮食与饲料工业,2016(2):69-73.DOI:10.7633/j.issn.1003-6202.2016.02.009.
[25]LIU P,LIANG S,WANG B J,et al.Development and validation of a sensitive LC-MS method for the determination of promethazine hydrochloride in human plasma and urine[J].European Journal of Drug Metabolism and Pharmacokinetics,2009,34(3/4):177-184.DOI:10.1007/BF03191171.
[26]程建桥,王传胜,张峰,等.超高效液相色谱-四极杆/静电场轨道阱高分辨质谱法测定猪肉中氯丙嗪及其代谢物[J].食品安全质量测试学报,2018,9(10):2440-2445.DOI:10.3969/j.issn.2095-0381.2018.10.027.
[27]李志刚,任南,马燕红,等.高效液相色谱-串联质谱法检测鲜冻肉中违禁注水药物[J].食品科学,2018,39(12):308-312.DOI:10.7506/spkx1002-6630-201812047.
[28]曹亚飞,康健,常巧英,等.QuEChERS结合液相色谱-串联质谱法快速测定奶酪中多种兽药残留[J].色谱,2015,33(2):132-139.DOI:10.3724/SP.J.1123.2014.10022.
[29]陈晓鹏,顾采琴,张施敬,等.QuEChERS-UPLC-MS/MS同时测定牛奶和奶粉中42种类固醇激素[J].食品科学,2018,39(8):314-321.DOI:10.7506/spkx1002-6630-201808049.
[30]CALATAYUD-VERNICH P,CALATAYUD F,SIMO E,et al.Efficiency of QuEChERS approach for determining 52 pesticide residues in honey and honey bees[J].MethodsX,2016(3):452-458.DOI:10.1016/j.mex.2016.05.005.
[31]LOZOWICKA B,RUTKOWSKA E,JANKOWSKA M.Influence of QuEChERS modifications on recovery and matrix effect during the multi-residue pesticide analysis in soil by GC/MS/MS and GC/ECD/NPD[J].Environmental Science and Pollution Research,2017,24(8):7124-7138.DOI:10.1007/s11356-016-8334-1.
[2]田鹏飞.国内外兽药企业创新与发展研究[D].武汉:华中农业大学,2012.
[3]闫小峰.浅谈我国兽药行业存在的主要问题及对策[J].中国兽药杂志,2009,43(8):35-38.DOI:10.3969/j.issn.1002-1280.2009.08.010.
[4]潘明飞,王俊平,方国臻,等.食品中农兽药残留检测新技术研究进展[J].食品科学,2014,35(15):277-282.DOI:10.7506/spkx1002-6630-201415056.
[5]车文军,张征,徐春祥,等.固相萃取/高效液相色谱法测定化妆品中5种抗组胺药物残留[J].分析测试学报,2012,31(8):1005-1008.DOI:10.3969/j.issn.1004-4957.2012.08.022.
[6]叶林虎,徐松,王丽莎,等.LC-MS/MS法检测皮肤科常用中药制剂中添加的7种抗组胺药物[J].药物分析杂志,2016,36(11):20-22.
[7]齐士林,吴敏,严丽娟,等.超高效液相色谱-质谱对动物源食品中氯丙嗪、异丙嗪及其代谢物的测定[J].分析测式学报,2009,28(6):677-681.DOI:10.3969/j.issn.1004-4957.2009.06.009.
[8]林芳,王一欣,李涛,等.安神类保健食品中新型非法添加药物的检测研究[J].食品安全质量检测学报,2016,7(4):1631-1636.
[9]邱畅,温预关,李恒,等.LC-ESI-MS/MS法测定人血浆中西替利嗪浓度及其咀嚼片人体生物等效性评价[J].中国临床药学杂志,2008,17(4):234-238.DOI:10.3969/j.issn.1007-4406.2008.04.012.
[10]王伟,陈振玲,崔玉静,等.超高效液相色谱质谱法同时测定人血浆中8种抗组胺类药物[J].中国临床药理学杂志,2017,33(10):929-932.DOI:10.13699/j.cnki.1001-6821.2017.10.017.
[11]卫生部,中国标准化管理委员会.饲料中盐酸异丙嗪、盐酸氯丙嗪、地西洋、硫酸硫利达嗪、奋乃静的同步测定高效液相色谱法和液相色谱质谱联用法:NY/T 1458-2007[S].北京:中国标准出版社,2007.
[12]YADOLLAH Y,MOHAMMAD F.Extraction and determination of trace amounts of chlorpromazine in biological fluids using magnetic solid phase extraction followed by HPLC[J].Journal of Pharmaceutical Analysis,2014,4(4):279-285.DOI:10.1016/j.jpha.2014.03.003.
[13]BROEDERS J,BLAAUBOER B,HERMENS J.Development of a negligible depletion-solid phase microextraetion method to determine the free concentration of chlorpromazine inaqueous samples containing albumin[J].Chromatography A,2011,1218:8529-8535.
[14]李婧妍,郭春锋,崔立辉,等.QuEChERs-HPLC-MS/MS法检测液态乳中9种β2-受体激动剂残留量[J].中国乳品工业,2016,44(11):49-52.DOI:10.3969/j.issn.1001-2230.2016.11.013.
[15]李琴,陶大利,周红,等.超高效液相-串联质谱法同时测定生鲜乳中45种兽药残留量[J].中国乳品工业,2016,44(1):44-47.DOI:10.3969/j.issn.1001-2230.2016.01.010.
[16]路平,曲志娜,谭维泉,等.气相色谱-质谱法测定猪肝中氯丙嗪残留的研究[J].中国动物检疫,2006,23(7):30-11.DOI:10.3969/j.issn.1005-944X.2006.07.016.
[17]吕燕,杨挺,赵健,等.气相色谱-质谱法测定猪肝中氯丙嗪残留量[J].分析试验室,2008,27(3):119-121.DOI:10.3969/j.issn.1000-0720.2008.03.033.
[18]WEJDAN S K,BAHRUDDIN S,NORMALIZA H A M,et al.Microsolid phase extraction with liquid chromatograph-tandem mass spectrometry for the determination of aflatoxins in coffee and malt beverage[J].Food Chemistry,2014,147(4):287-294.DOI:10.1016/j.foodchem.2013.09.049.
[19]黄德凤,VAN TRAN K,MICHEAL S,等.Oasis PRIME HLB-UPLC-MS/MS测定猪肉中78种兽药残留[G]//Oasis PRIME HLB食品应用文集.2015:8-10.
[20]LI M,KONG W,LI Y,et al.High-throughput determination of multimycotoxins in Chinese yamand related products by ultra fast liquid chromatography coupled with tandem mass spectrometry after onestep extraction[J].Journal of Chromatography B,2016,1022(6):118-125.DOI:10.1016/j.jchromb.2016.04.014.
[21]孙伟华,杨欢,曹赵云,等.基于分散固相萃取液相色谱-串联质谱法测定大米中8种真菌毒素[J].分析测试学报,2017,36(1):47-53.DOI:10.3969/j.issn.1004-4957.2017.01.008.
[22]李诗言,柯庆青,王鼎南,等.QuEChERS-LC-MS/MS法测定水产配合饲料中4种常见黄曲霉毒素残留[J].中国渔业质量与标准,2017,7(5):25-31.DOI:10.3696/j.issn.2095-1833.2017.05.004.
[23]谢寒冰,蒋万枫,赵海峰,等.高效液相色谱法-四极杆飞行时间质谱法测定猪肉中20种兽药残留量[J].理化检验:化学分册,2014,50(6):702-707.
[24]梁玉禧,李益珍,郭小敏,等.液质联用法测定猪肉产品中违禁添加物氯丙嗪、异氯丙嗪和万古霉素残留研究[J].粮食与饲料工业,2016(2):69-73.DOI:10.7633/j.issn.1003-6202.2016.02.009.
[25]LIU P,LIANG S,WANG B J,et al.Development and validation of a sensitive LC-MS method for the determination of promethazine hydrochloride in human plasma and urine[J].European Journal of Drug Metabolism and Pharmacokinetics,2009,34(3/4):177-184.DOI:10.1007/BF03191171.
[26]程建桥,王传胜,张峰,等.超高效液相色谱-四极杆/静电场轨道阱高分辨质谱法测定猪肉中氯丙嗪及其代谢物[J].食品安全质量测试学报,2018,9(10):2440-2445.DOI:10.3969/j.issn.2095-0381.2018.10.027.
[27]李志刚,任南,马燕红,等.高效液相色谱-串联质谱法检测鲜冻肉中违禁注水药物[J].食品科学,2018,39(12):308-312.DOI:10.7506/spkx1002-6630-201812047.
[28]曹亚飞,康健,常巧英,等.QuEChERS结合液相色谱-串联质谱法快速测定奶酪中多种兽药残留[J].色谱,2015,33(2):132-139.DOI:10.3724/SP.J.1123.2014.10022.
[29]陈晓鹏,顾采琴,张施敬,等.QuEChERS-UPLC-MS/MS同时测定牛奶和奶粉中42种类固醇激素[J].食品科学,2018,39(8):314-321.DOI:10.7506/spkx1002-6630-201808049.
[30]CALATAYUD-VERNICH P,CALATAYUD F,SIMO E,et al.Efficiency of QuEChERS approach for determining 52 pesticide residues in honey and honey bees[J].MethodsX,2016(3):452-458.DOI:10.1016/j.mex.2016.05.005.
[31]LOZOWICKA B,RUTKOWSKA E,JANKOWSKA M.Influence of QuEChERS modifications on recovery and matrix effect during the multi-residue pesticide analysis in soil by GC/MS/MS and GC/ECD/NPD[J].Environmental Science and Pollution Research,2017,24(8):7124-7138.DOI:10.1007/s11356-016-8334-1.