超高效液相色谱-串联质谱法测定罗非鱼血浆和肌肉中的丁香酚残留
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摘要
建立了一种使用超高效液相色谱-串联质谱联用仪检测罗非鱼血浆和肌肉中丁香酚残留的方法。血浆经甲醇提取,以乙腈和0.1%的氨水为流动相,采用梯度洗脱对待测物进行分离,采用电喷雾离子源(ESI)负离子扫描,多反应监测模式(MRM)测定,外标法定量;肌肉经正己烷提取,HLB固相萃取柱净化,内标法定量。结果表明,在1.0～100.0μg/L线性范围内,丁香酚在罗非鱼血浆中线性良好(r=0.998 5),平均回收率为97.0%～98.9%,相对标准偏差为1.14%～2.21%;在1.0～90.0μg/kg线性范围内,丁香酚在罗非鱼肌肉中线性良好(r=0.999 8),平均回收率为90.2%～95.9%,相对标准偏差为1.91%～5.28%。血浆和肌肉中丁香酚的定量限分别为2.0μg/L和2.0μg/kg。实验结果满足罗非鱼血浆和肌肉中丁香酚残留的检测需求,可为日后丁香酚相关检测研究提供参考。
The study aims to establish a method for the determination of eugenol residues in the plasma and muscle of tilapia using ultra performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS). The plasma was directly extracted with methanol. The muscle was extracted with n-hexane and purified by hydrophilic-lipophilic balance(HLB) cartridge. Both of them were separated by a mobile phase of acetonitrile and 0.1% ammonia using gradient elution, and detected by tandem mass spectrometry using multiple reaction monitoring(MRM) in negative ion mode. The plasma sample was determined by an external reference method, and the muscle sample was quantitated with an internal standard method. The results showed that the eugenol from the plasma of tilapia had a good linear relationship(r=0.998 5)in the range of 1.0-100.0 μg/L, the average recovery was 97.0%-98.9%, and the relative standard deviation was 1.14%-2.21%. The eugenol from in the muscle of tilapia had a good linear relationship in the range of 1.0-90.0 μg/kg(r=0.999 8), the average recovery was 90.2%-95.9%, and the relative standard deviation was 1.91%-5.28%. The limits of quantity(LOQs) were 2.0 μg/L in plasma and 2.0 μg/kg in muscle, respectively.It suggested that the established method can meet the requirement for the detection of eugenol residues in the plasma and muscle of tilapia, which could provide reference for relevant studies on the detection of eugenol in future.
The study aims to establish a method for the determination of eugenol residues in the plasma and muscle of tilapia using ultra performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS). The plasma was directly extracted with methanol. The muscle was extracted with n-hexane and purified by hydrophilic-lipophilic balance(HLB) cartridge. Both of them were separated by a mobile phase of acetonitrile and 0.1% ammonia using gradient elution, and detected by tandem mass spectrometry using multiple reaction monitoring(MRM) in negative ion mode. The plasma sample was determined by an external reference method, and the muscle sample was quantitated with an internal standard method. The results showed that the eugenol from the plasma of tilapia had a good linear relationship(r=0.998 5)in the range of 1.0-100.0 μg/L, the average recovery was 97.0%-98.9%, and the relative standard deviation was 1.14%-2.21%. The eugenol from in the muscle of tilapia had a good linear relationship in the range of 1.0-90.0 μg/kg(r=0.999 8), the average recovery was 90.2%-95.9%, and the relative standard deviation was 1.91%-5.28%. The limits of quantity(LOQs) were 2.0 μg/L in plasma and 2.0 μg/kg in muscle, respectively.It suggested that the established method can meet the requirement for the detection of eugenol residues in the plasma and muscle of tilapia, which could provide reference for relevant studies on the detection of eugenol in future.
引文
[1] 吕海燕,王群,刘欢,等.鱼用麻醉剂安全性研究进展[J].中国渔业质量与标准,2013(2):24-28.
[2] Meinertz J R,Schreier T M.Depletion of isoeugenol residues from the fillet tissue of AQUI-STM exposed rainbow trout(Oncorhynchus mykiss) [J].Aquaculture,2009,296(3/4):200-206.
[3] 柯常亮,刘奇,陈浩文,等.气相色谱-串联质谱法测定水中丁香酚残留 [J].中国渔业质量与标准,2014,4(4):49-55.
[4] Executive summary of safety and toxicology information﹣isoeugenol[R].1991.https://ntp.niehs.nih.gov/ntp/htdocs/chem_backgronnd/exsumpdf/isoeugeno1_508.pdf.
[5] U.S.National Institutes of Health.Nationaltoxicologyprogram technical report series,No.223 carcinogenesis studies of eugenol[EB/OL].(1983-12-16) [2018-05-30].http://ntp.niehs.nih.gov/ntp/htdocs/It_ rpts/tr223.pdf.
[6] US Food and Drug Administration.Guideline No.150.guidance for industry:concerns related to the use of clove oil as an anesthetic for fish[EB/OL].(2007-04-24)[20 18-05-30].http://www.Fda.gov/downloads/animalveterinary/guidancecomplianceenforcement/guidanceforindustry/ucm052520.pdf.
[7] Canadian Council on Animal Care.Guidelines for additional information on effects of euthanasia methods on research results:clove oi1(eugenol)[EB/OL].(2016-04-16)[2018-05-30].http://www.ccac.ca/en_/standards/guidelines/additional/additional-information-on-effects-of-euthanasia-methods.on-research-results/clove-oil.
[8] The Japan Food Chemical Research Foundation.Revision of MRLs of agricultural chemicals,feed additives and veterinary drugs in foods[EB/OL].(2016-04-20)[2018-05-30].http://www.ffcr.or.jp/zaidan/ffcrhome.nsf/TrueMainE?OpenFrameSet.
[9] 赵东豪,王强,王旭峰,等.超高效液相色谱-串联质谱法检测鱼肉和暂养水中丁香酚残留量[J].食品科学,2016,37(24):252-256.
[10] 《生命时报》.鲜鱼被曝注“丁香油水门汀”麻醉后运输[EB/OL].(2013-03-18)[2018-05-30].http://health.qq.com/a/20130318/000035.htm.
[11] 李晋成,刘欢,王群,等.水产品中丁香酚的残留风险评估研究进展[J].食品安全质量检测学报,2014,5(11):3598-3602.
[12] 陈焕,黄和,高平,等.高效液相色谱法测定水产品中丁香酚类麻醉剂的残留量[J].食品与发酵工业,2014,40(12):156-160.
[13] 高平,黄和,刘文侠,等.固相萃取一高效液相色谱一荧光检测法测定水产品中4种丁香酚类化合物[J].中国食品卫生杂志,2016,28(1):56-61.
[14] 黄武,徐金龙,刘建芳,等.高效液相色谱法检测罗非鱼中丁香酚残留量[J].食品安全质量检测学报,2018,9(1):103-106.
[15] 焦亚琴,吕世明,谭艾娟,等.高效液相色谱法测定鲤鱼血浆中丁香酚含量[J].基因组学与应用生物学,2016(8):2022-2026.
[16] 孙宇航,王绿洲,李锋刚,等.丁香酚在罗非鱼体内的药物代谢动力学研究[J].水产科技情报,2018,45(4):218-222.
[17] 陈焕,黄和,高平,等.分散固相萃取-气相色谱法同时测定水产品中六种丁香酚类麻醉剂的残留量[J].食品工业科技,2015,36(8):88-92.
[18] 孙鹏,高玉玲,王金梅,等.漂浮固化分散液液微萃取-UHPLC-MS/MS法测定水产品中丁香酚的残留量[J].质谱学报,2017,38(3):342-348.
[19] 倪峥飞,顾晔,冯永巍,等.UPLC-MS/MS 同时测定水产品中4种丁香酚类麻醉剂残留量[J].质谱学报,2018,39(4):451-458.
[20] 芦智远,刘辰乾,冯歆轶,等.超高效液相色谱-串联质谱法鱼肉丁香酚的残留量测定[J].科学养鱼,2016 (7):76-77.
[21] 陈焕,黄和,高平,等.GC-MS法同时测定水产品中6种丁香酚类麻醉剂的残留量[J].食品工业,2017,38(7):305-309.
[22] 余颖.气相色谱一质谱法测定鱼肉中丁香酚的残留[J].福州大学学报(自然科学版),2015,43(2):266-270.
[23] Li J,Zhang J,Liu Y.Optimization of solid-phase-extraction cleanup and validation of quantitative determination of eugenol in fish samples by gas chromatography-tandem mass spectrometry [J].Anal Bioanal Chem,2015,407(21):6563- 6568.
[24] 刘海新,余颖,汤水粉,等.气相色谱质谱法测定养殖水体中丁香酚含量不确定度评定[J].渔业研究,2016,38(4):310 - 317.
[25] Li J,Liu H,Wang C,et al.Determination of eugenol in fish and shrimp muscle tissue by stable isotope dilution assay and solid-phase extraction coupled gas chromatography-triple quadrupole mass spectrometry [J].Anal Bioanaly Chem,2016,408(24):6537-6544.
[2] Meinertz J R,Schreier T M.Depletion of isoeugenol residues from the fillet tissue of AQUI-STM exposed rainbow trout(Oncorhynchus mykiss) [J].Aquaculture,2009,296(3/4):200-206.
[3] 柯常亮,刘奇,陈浩文,等.气相色谱-串联质谱法测定水中丁香酚残留 [J].中国渔业质量与标准,2014,4(4):49-55.
[4] Executive summary of safety and toxicology information﹣isoeugenol[R].1991.https://ntp.niehs.nih.gov/ntp/htdocs/chem_backgronnd/exsumpdf/isoeugeno1_508.pdf.
[5] U.S.National Institutes of Health.Nationaltoxicologyprogram technical report series,No.223 carcinogenesis studies of eugenol[EB/OL].(1983-12-16) [2018-05-30].http://ntp.niehs.nih.gov/ntp/htdocs/It_ rpts/tr223.pdf.
[6] US Food and Drug Administration.Guideline No.150.guidance for industry:concerns related to the use of clove oil as an anesthetic for fish[EB/OL].(2007-04-24)[20 18-05-30].http://www.Fda.gov/downloads/animalveterinary/guidancecomplianceenforcement/guidanceforindustry/ucm052520.pdf.
[7] Canadian Council on Animal Care.Guidelines for additional information on effects of euthanasia methods on research results:clove oi1(eugenol)[EB/OL].(2016-04-16)[2018-05-30].http://www.ccac.ca/en_/standards/guidelines/additional/additional-information-on-effects-of-euthanasia-methods.on-research-results/clove-oil.
[8] The Japan Food Chemical Research Foundation.Revision of MRLs of agricultural chemicals,feed additives and veterinary drugs in foods[EB/OL].(2016-04-20)[2018-05-30].http://www.ffcr.or.jp/zaidan/ffcrhome.nsf/TrueMainE?OpenFrameSet.
[9] 赵东豪,王强,王旭峰,等.超高效液相色谱-串联质谱法检测鱼肉和暂养水中丁香酚残留量[J].食品科学,2016,37(24):252-256.
[10] 《生命时报》.鲜鱼被曝注“丁香油水门汀”麻醉后运输[EB/OL].(2013-03-18)[2018-05-30].http://health.qq.com/a/20130318/000035.htm.
[11] 李晋成,刘欢,王群,等.水产品中丁香酚的残留风险评估研究进展[J].食品安全质量检测学报,2014,5(11):3598-3602.
[12] 陈焕,黄和,高平,等.高效液相色谱法测定水产品中丁香酚类麻醉剂的残留量[J].食品与发酵工业,2014,40(12):156-160.
[13] 高平,黄和,刘文侠,等.固相萃取一高效液相色谱一荧光检测法测定水产品中4种丁香酚类化合物[J].中国食品卫生杂志,2016,28(1):56-61.
[14] 黄武,徐金龙,刘建芳,等.高效液相色谱法检测罗非鱼中丁香酚残留量[J].食品安全质量检测学报,2018,9(1):103-106.
[15] 焦亚琴,吕世明,谭艾娟,等.高效液相色谱法测定鲤鱼血浆中丁香酚含量[J].基因组学与应用生物学,2016(8):2022-2026.
[16] 孙宇航,王绿洲,李锋刚,等.丁香酚在罗非鱼体内的药物代谢动力学研究[J].水产科技情报,2018,45(4):218-222.
[17] 陈焕,黄和,高平,等.分散固相萃取-气相色谱法同时测定水产品中六种丁香酚类麻醉剂的残留量[J].食品工业科技,2015,36(8):88-92.
[18] 孙鹏,高玉玲,王金梅,等.漂浮固化分散液液微萃取-UHPLC-MS/MS法测定水产品中丁香酚的残留量[J].质谱学报,2017,38(3):342-348.
[19] 倪峥飞,顾晔,冯永巍,等.UPLC-MS/MS 同时测定水产品中4种丁香酚类麻醉剂残留量[J].质谱学报,2018,39(4):451-458.
[20] 芦智远,刘辰乾,冯歆轶,等.超高效液相色谱-串联质谱法鱼肉丁香酚的残留量测定[J].科学养鱼,2016 (7):76-77.
[21] 陈焕,黄和,高平,等.GC-MS法同时测定水产品中6种丁香酚类麻醉剂的残留量[J].食品工业,2017,38(7):305-309.
[22] 余颖.气相色谱一质谱法测定鱼肉中丁香酚的残留[J].福州大学学报(自然科学版),2015,43(2):266-270.
[23] Li J,Zhang J,Liu Y.Optimization of solid-phase-extraction cleanup and validation of quantitative determination of eugenol in fish samples by gas chromatography-tandem mass spectrometry [J].Anal Bioanal Chem,2015,407(21):6563- 6568.
[24] 刘海新,余颖,汤水粉,等.气相色谱质谱法测定养殖水体中丁香酚含量不确定度评定[J].渔业研究,2016,38(4):310 - 317.
[25] Li J,Liu H,Wang C,et al.Determination of eugenol in fish and shrimp muscle tissue by stable isotope dilution assay and solid-phase extraction coupled gas chromatography-triple quadrupole mass spectrometry [J].Anal Bioanaly Chem,2016,408(24):6537-6544.