海产贝类脂溶性贝毒素的高效液相色谱-串联质谱分析方法及其食用安全性评价
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摘要
针对海产贝类存在多种脂溶性贝毒素复合污染的现状,采用高效液相色谱-串联质谱联用技术(HPLC-MS/MS)对海产贝类中的常见脂溶性贝毒素进行同步检测,结合多种毒素复合污染的风险评估方法,用于市售海产贝类的食用安全风险评价。结果表明,在选定的实验条件下,8种典型脂溶性贝毒素加标回收率在63.2%~88.8%之间,方法的精密度(相对标准偏差(RSD)≤14.5%)和灵敏度(检出限为0.5~2.7 ng/g)良好,能满足海产贝类样品的检测要求。在采集的105个市售海产贝类样品中,42.86%的样品中至少检出了一种脂溶性贝毒素,其中鳍藻毒素-1(DTX1)的含量均值最高,为47.6μg/kg,对海产贝类污染最严重。根据每日人均贝类摄入量(TDI)和各种脂溶性贝毒素的急性中毒参考剂量(ARf D),通过计算综合风险指数∑ERI进行市售海产贝类食用安全性评价,结果表明,在所检测的样品中,存在食用安全隐患和高风险的市售海产贝类比率为19.05%,其中扇贝的食用安全风险最大。本研究建立的基于海产贝类中脂溶性贝毒素物质组复合污染的风险评价方法,与欧盟的海产品贝毒素限量标准评价方法(单指标法)相比更加严格,可以使贝类食用者更好地规避中毒风险。
In view of the present situation that edible marine shellfishes are combinedly contaminated by different kinds of lipophilic toxins,common lipophilic shellfish toxins in marine shellfishes were simultaneously detected by liquid chromatography-tandem mass spectrometry,and the safety risk of commercial marine shellfish was evaluated using the risk assessment method based on combined contamination of various toxins.Under the optimum conditions, satisfactory recoveries( 63. 3%-88. 8%), precision( relative standard deviations RSD≤14.5%) and sensitivity( limit of detection in the range of 0.5-2.7 ng/g) of the method were achieved for all the analytes. Among the 105 commercially available shellfish samples,42.86% of the samples had at least a kind of toxin. The highest average content was 47.6 μg/kg of DTX1,which was the most serious contaminant for marine shellfishes. The total Expose Risk Index( ΣERI) was calculated based on Tolerable Daily Intake( TDI) and Acute Reference Dose( ARf D) of each toxin to evaluate the safety risk of commercial marine shellfish. The results showed that the ratio of commercially available marine shellfish with safety risk was 19.05% and the food safety risk of scallop was the highest. In summary,a new method based on the combined contamination of lipophilic shellfish toxins was successfully developed for risk assessment of the commercial marine shellfish. The proposed method is more harsh compared with the European Food Safety Authority( EFSA) regulation and can make shellfish consumers better to avoid the risk of poisoning.
In view of the present situation that edible marine shellfishes are combinedly contaminated by different kinds of lipophilic toxins,common lipophilic shellfish toxins in marine shellfishes were simultaneously detected by liquid chromatography-tandem mass spectrometry,and the safety risk of commercial marine shellfish was evaluated using the risk assessment method based on combined contamination of various toxins.Under the optimum conditions, satisfactory recoveries( 63. 3%-88. 8%), precision( relative standard deviations RSD≤14.5%) and sensitivity( limit of detection in the range of 0.5-2.7 ng/g) of the method were achieved for all the analytes. Among the 105 commercially available shellfish samples,42.86% of the samples had at least a kind of toxin. The highest average content was 47.6 μg/kg of DTX1,which was the most serious contaminant for marine shellfishes. The total Expose Risk Index( ΣERI) was calculated based on Tolerable Daily Intake( TDI) and Acute Reference Dose( ARf D) of each toxin to evaluate the safety risk of commercial marine shellfish. The results showed that the ratio of commercially available marine shellfish with safety risk was 19.05% and the food safety risk of scallop was the highest. In summary,a new method based on the combined contamination of lipophilic shellfish toxins was successfully developed for risk assessment of the commercial marine shellfish. The proposed method is more harsh compared with the European Food Safety Authority( EFSA) regulation and can make shellfish consumers better to avoid the risk of poisoning.
引文
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3 Gerssen A,Mulder P P,De B J.Anal.Chim.Acta,2011,685(2):176-185
4 LIU Ren-Yan,LIU Lei,LIANG Yu-Bo,YU Ji,XU Dao-Yan,WEI Ning,YANG Lin,GUO Hao.Marine.Environ.Sci.,2016,35(5):787-800刘仁沿,刘磊,梁玉波,于姬,许道艳,韦宁,杨琳,郭浩.海洋环境科学,2016,35(5):787-800
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8 ZHAO Qian-Cheng,LAI Peng,QIN Cheng,WANG Chong,WU Bin,LIU Di,CHANG Ya-Qing.Food.Sci.Technol.,2009,(11):287-292赵前程,来鹏,秦成,王翀,吴斌,刘迪,常亚青.食品科技,2009,(11):287-292
9 Lee J S,Yanagi T,Kenma R,Yasumoto T.Agr.Biol.Chem.,1987,51(3):877-881
10 Gerssen A,Mulder P P,Mcelhinney M A,De B J.J.Chromatogr.A,2009,1216(9):1421-1430
11 Regueiro J,Rossignoli A E,Alvarez G,Blanco J.Food Chem.,2011,129(2):533-540
12 CHEN Jian-Gang,ZHU Bing-Hui,LIANG Su-Dan,ZHANG Gui,WU Xi-Mei.Chinese J.Food Hygiene,2015,27(6):624-629陈剑刚,朱炳辉,梁素丹,张瑰,吴西梅.中国食品卫生杂志,2015,27(6):624-629
13 TAN Zhi-Jun,WU Hai-Yan,GUO Meng-Meng,YANG Fan,WANG Lian-Zhu,LI Zhao-Xin,ZHAI Yu-Xiu.J.Fishery Sci.,2013,20(2):467-479谭志军,吴海燕,郭萌萌,杨帆,王联珠,李兆新,翟毓秀.中国水产科学,2013,20(2):467-479
14 Commission decision 2002/225/EC of 15 March 2002 laying down detailed rules for the implementation of Council Directive 91 /492/EEC as regards the maximum levels and the methods of analysis of certain marine biotoxins in bivalve molluscs,echino-derms,tunicates and marine gastropods.Off.J.Eur.Communities,2002:62-63
15 Regulation(EC)No 853/2004 of the European Parliament and of the Council of 29April 2004 laying down specific hygiene rules for food of animal origin.Off.J.Eur.Union,2004,L226:22-82
16 YU Hong,LI Xiao-Jing,LIU Miao,XIE Jin,PENG Rong-Fei.Chinese J.Health.Lab.Technol.,2014,(18):2702-2704于鸿,李晓晶,刘苗,谢进,彭荣飞.中国卫生检验杂志,2014,(18):2702-2704
17 Munday R,Reeve J.Toxins,2013,5(11):2109-2137
18 Lawrence J,Loreal H,Toyofuku H,Hess P,Iddya K,Ababouch L.Assessment and Management of Biotoxin Risks in Bivalve Molluscs.FAO Fisheries and Aquaculture.Technical Paper No.551,Rome:FAO,2011:43-44
19 Henshilwood K,Deegan B,Mcmahon T,Cusack C,Keaveney S,Silke J,Ocinneide M,Lyons D,Hess P.Proceedings of the 5th International conference on Molluscan Shellfish Safety,Ireland:Galway,2006:464-468
20 Munday R,Aune T,Rossini G P.Toxicology of the Yessotoxins,Florida:Taylor&Fran CI,2008:329-339
21 Munday R,Towers N R,Mackenzie L,Beuzenberg V,Holland P T,Miles C O.Toxicon,2004,44(2):173-178
22 Li X,Li Z Y,Chen J H,Shi Q,Zhang R T,Wang S,Wang X R.Chemosphere,2014,111:560-567
23 WANG Yan-Long,CHEN Jun-Hui,GAO Li-Yuan,WANG Shuai,ZHENG Xiao-Ling,SUN Cheng-Jun,WANG Xiao-Ru.Chinese J.Anal.Chem.,2016,44(3):335-341王艳龙,陈军辉,高莉媛,王帅,郑晓玲,孙承君,王小如.分析化学,2016,44(3):335-341
24 Wang Y L,Chen J H,Li Z Y,Wang S,Shi Q,Cao W,Zheng X L,Sun C J,Wang X R,Zheng L.Mar.Pollut.Bull.,2015,101(2):954-960
25 MU Qing-Lin,FANG Jie,WAN Han-Xing,WANG Xiao-Hua,CAO Liu-Yan,ZHANG Qing-Hong.Chinese J.Anal.Chem.,2011,39(1):111-114母清林,方杰,万汉兴,王晓华,曹柳燕,张庆红.分析化学,2011,39(1):111-114
26 GB 5009.212-2016,Determination of Lipophilic Shellfish Toxins in Shellfish.National Standards of the People's Republic of China贝类中腹泻性贝类毒素的测定.中华人民共和国国家标准.GB 5009.212-2016
27 YAO Jian-Hua,TAN Zhi-Jun,ZHOU De-Qing.Chinese J.Anal.Chem.,2010,38(12):1714-1720姚建华,谭志军,周德庆.分析化学,2010,38(12):1714-1720
28 SUN Xing-Quan,ZHENG Qiu-Yue,PANG Yan-Hua,LI Yi-Chen,XIAO Shan-Shan,XU Jing,CAO Ji-Juan.Chinese J.Anal.Chem,2013,41(9):1423-1427孙兴权,郑秋月,庞艳华,李一尘,肖珊珊,徐静,曹际娟.分析化学,2013,41(9):1423-1427
29 DENG Shuo,YU Wei-Sen,LYU Xiao-Jing,TAN Zhi-Jun.Prev.Med.Trib.,2017,(1):11-16邓硕,于维森,吕晓静,谭志军.预防医学论坛,2017,(1):11-16
30 Alexander J,Benford D,Boobis A,Ceccatelli S,Cravedi J P,Domenico A D,Doerge D,Dogliotti E,Edler L,Farmer P,Filipic M,Fink-Gremmels J,Furst P,Guerin T,Knutsen H K,Machala M,Mutti A,Schlatter J,Leeuwen R,Verger P.The EFSA J.,2009,1306:1-23
2 LIU Ren-Yan,GAO Chun-Lei,LIANG Yu-Bo,ZHANG Fang,LIU Yong-Jian,XU Dao-Yan.Acta Oceanol.Sin.,2008,30(6):171-176刘仁沿,高春蕾,梁玉波,张芳,刘永健,许道艳.海洋学报,2008,30(6):171-176
3 Gerssen A,Mulder P P,De B J.Anal.Chim.Acta,2011,685(2):176-185
4 LIU Ren-Yan,LIU Lei,LIANG Yu-Bo,YU Ji,XU Dao-Yan,WEI Ning,YANG Lin,GUO Hao.Marine.Environ.Sci.,2016,35(5):787-800刘仁沿,刘磊,梁玉波,于姬,许道艳,韦宁,杨琳,郭浩.海洋环境科学,2016,35(5):787-800
5 Li A F,Ma J G,Cao J J,Mc Carron P.Toxicon,2012,60(3):420-425
6 ZHONG Yuan-Fang,JIANG Hai-Tang,LI Yun-Feng,CHEN Rong-Kai,LIN Wei.Chinese.Prim.Heal.Care,2014,28(2):104-105钟苑芳,江海棠,李云峰,陈荣凯,林维.中国初级卫生保健,2014,28(2):104-105
7 Yasumoto T,Oshima Y,Yamaguchi M.Nippon Suisan Gakkaishi,1978,44(11):1249-1255
8 ZHAO Qian-Cheng,LAI Peng,QIN Cheng,WANG Chong,WU Bin,LIU Di,CHANG Ya-Qing.Food.Sci.Technol.,2009,(11):287-292赵前程,来鹏,秦成,王翀,吴斌,刘迪,常亚青.食品科技,2009,(11):287-292
9 Lee J S,Yanagi T,Kenma R,Yasumoto T.Agr.Biol.Chem.,1987,51(3):877-881
10 Gerssen A,Mulder P P,Mcelhinney M A,De B J.J.Chromatogr.A,2009,1216(9):1421-1430
11 Regueiro J,Rossignoli A E,Alvarez G,Blanco J.Food Chem.,2011,129(2):533-540
12 CHEN Jian-Gang,ZHU Bing-Hui,LIANG Su-Dan,ZHANG Gui,WU Xi-Mei.Chinese J.Food Hygiene,2015,27(6):624-629陈剑刚,朱炳辉,梁素丹,张瑰,吴西梅.中国食品卫生杂志,2015,27(6):624-629
13 TAN Zhi-Jun,WU Hai-Yan,GUO Meng-Meng,YANG Fan,WANG Lian-Zhu,LI Zhao-Xin,ZHAI Yu-Xiu.J.Fishery Sci.,2013,20(2):467-479谭志军,吴海燕,郭萌萌,杨帆,王联珠,李兆新,翟毓秀.中国水产科学,2013,20(2):467-479
14 Commission decision 2002/225/EC of 15 March 2002 laying down detailed rules for the implementation of Council Directive 91 /492/EEC as regards the maximum levels and the methods of analysis of certain marine biotoxins in bivalve molluscs,echino-derms,tunicates and marine gastropods.Off.J.Eur.Communities,2002:62-63
15 Regulation(EC)No 853/2004 of the European Parliament and of the Council of 29April 2004 laying down specific hygiene rules for food of animal origin.Off.J.Eur.Union,2004,L226:22-82
16 YU Hong,LI Xiao-Jing,LIU Miao,XIE Jin,PENG Rong-Fei.Chinese J.Health.Lab.Technol.,2014,(18):2702-2704于鸿,李晓晶,刘苗,谢进,彭荣飞.中国卫生检验杂志,2014,(18):2702-2704
17 Munday R,Reeve J.Toxins,2013,5(11):2109-2137
18 Lawrence J,Loreal H,Toyofuku H,Hess P,Iddya K,Ababouch L.Assessment and Management of Biotoxin Risks in Bivalve Molluscs.FAO Fisheries and Aquaculture.Technical Paper No.551,Rome:FAO,2011:43-44
19 Henshilwood K,Deegan B,Mcmahon T,Cusack C,Keaveney S,Silke J,Ocinneide M,Lyons D,Hess P.Proceedings of the 5th International conference on Molluscan Shellfish Safety,Ireland:Galway,2006:464-468
20 Munday R,Aune T,Rossini G P.Toxicology of the Yessotoxins,Florida:Taylor&Fran CI,2008:329-339
21 Munday R,Towers N R,Mackenzie L,Beuzenberg V,Holland P T,Miles C O.Toxicon,2004,44(2):173-178
22 Li X,Li Z Y,Chen J H,Shi Q,Zhang R T,Wang S,Wang X R.Chemosphere,2014,111:560-567
23 WANG Yan-Long,CHEN Jun-Hui,GAO Li-Yuan,WANG Shuai,ZHENG Xiao-Ling,SUN Cheng-Jun,WANG Xiao-Ru.Chinese J.Anal.Chem.,2016,44(3):335-341王艳龙,陈军辉,高莉媛,王帅,郑晓玲,孙承君,王小如.分析化学,2016,44(3):335-341
24 Wang Y L,Chen J H,Li Z Y,Wang S,Shi Q,Cao W,Zheng X L,Sun C J,Wang X R,Zheng L.Mar.Pollut.Bull.,2015,101(2):954-960
25 MU Qing-Lin,FANG Jie,WAN Han-Xing,WANG Xiao-Hua,CAO Liu-Yan,ZHANG Qing-Hong.Chinese J.Anal.Chem.,2011,39(1):111-114母清林,方杰,万汉兴,王晓华,曹柳燕,张庆红.分析化学,2011,39(1):111-114
26 GB 5009.212-2016,Determination of Lipophilic Shellfish Toxins in Shellfish.National Standards of the People's Republic of China贝类中腹泻性贝类毒素的测定.中华人民共和国国家标准.GB 5009.212-2016
27 YAO Jian-Hua,TAN Zhi-Jun,ZHOU De-Qing.Chinese J.Anal.Chem.,2010,38(12):1714-1720姚建华,谭志军,周德庆.分析化学,2010,38(12):1714-1720
28 SUN Xing-Quan,ZHENG Qiu-Yue,PANG Yan-Hua,LI Yi-Chen,XIAO Shan-Shan,XU Jing,CAO Ji-Juan.Chinese J.Anal.Chem,2013,41(9):1423-1427孙兴权,郑秋月,庞艳华,李一尘,肖珊珊,徐静,曹际娟.分析化学,2013,41(9):1423-1427
29 DENG Shuo,YU Wei-Sen,LYU Xiao-Jing,TAN Zhi-Jun.Prev.Med.Trib.,2017,(1):11-16邓硕,于维森,吕晓静,谭志军.预防医学论坛,2017,(1):11-16
30 Alexander J,Benford D,Boobis A,Ceccatelli S,Cravedi J P,Domenico A D,Doerge D,Dogliotti E,Edler L,Farmer P,Filipic M,Fink-Gremmels J,Furst P,Guerin T,Knutsen H K,Machala M,Mutti A,Schlatter J,Leeuwen R,Verger P.The EFSA J.,2009,1306:1-23