麦鲮(Cirrhina mrigola)肌肉组织在孔雀石绿阳性环境中的残留消除规律
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摘要
为研究在使用过孔雀石绿(malachite green,MG)的池塘中养殖麦鲮(Cirrhina mrigola)是否会对其产生一定的影响,连续两年度开展MG阳性环境下养殖麦鲮代谢残留实验。首年6月,将初始体重为(2. 68±0. 51) g的麦鲮放在MG阳性环境中饲养,于养殖后的0、0. 5、1、2、3、5、10、15、20和30 d取样;次年6月,重新换水后,再次放入与首年体长、质量相近的麦鲮进行饲养,于放养后0、1、2、5、10、15、20、30、40、50、70、90、120、150、180和210 d取样。采用高效液相色谱—串联质谱法(HPLC-MS/MS)测定麦鲮肌肉中MG及其代谢物无色孔雀石绿(leucomalachite green,LMG)的残留量,评估阳性环境中MG在放养鲮肌肉中残留情况。结果显示:1)建立的高效液相色谱-串联质谱方法检测MG和LMG在1. 0～200. 0 ng/m L质量浓度范围内分别呈现良好的线性关系,MG和LMG的检出限(LOD)与定量限(LOQ)均为0. 5μg/kg。2)首次放养0～30 d期间,肌肉中MG残留量先随时间波动式上升后逐渐下降,至160. 00μg/kg左右达动态平衡;同时肌肉中LMG残留量以同样的趋势至3 370. 00μg/kg左右,并达动态平衡。3)次年相同池塘再次放养与首年体长、质量相近的麦鲮后,0～70 d期间,其体内MG一直维持在(1. 43±2. 45)～(4. 98±2. 01)μg/kg,直到90 d后未检出; LMG在0～5 d残留量从(3. 10±2. 31)μg/kg上升到(105. 30±8. 25)μg/kg,随后波动式下降,直至180 d低于定量限(0. 50μg/kg)。上述结果表明,在使用过MG的养殖池塘中放养麦鲮,能够连续两年度在其肌肉中检出MG和LMG,且第一年度的检出值远高于第二年度的检出值,说明阳性池塘环境中MG和LMG难以短时间内彻底消除,能够在放养麦鲮肌肉中残留富集。本研究可为监督和执法管理部门对水产品中MG来源调查提供参考依据,对水产品质量安全风险评估具有一定意义。
To study on enrichment effect of mud carp( Cirrhina mrigola) in malachite green( MG) positive environment and the elimination rules of malachite green in vivo,the mud carp with body weight of( 2. 68 ± 0. 51) g were fed in malachite green positive environment( ponds splashed with MG before) in the first year of June,and sampling at 0,0. 5,1,2,3,5,10,15,20 and 30 d after breeding. After changing breeding water in the next year of June,the mud crap,with similar body length and quality to previous samples,sampling at 0, 1, 2, 5, 10, 15, 20, 30, 40,50, 70,90, 120,150,180 and 210 d after feeding in the same pond. The residues of MG and LMG in fish muscles were determined by high performance liquid chromatography tandem mass spectrometry( HPLC-MS/MS) in order to evaluate the residual MG and LMG in stocking muscles in MG positive environment. The results showed that: 1) the established high performance liquid chromatography-tandem mass spectrometry method showed that MG and LMG had a good linear relationship in the range of 1. 0 ～ 200. 0 ng/m L respectively. The detection limits( LOD) and limit of quantitation( LOQ) of MG and LMG were both 0. 5 μg/kg. 2) during the first feeding 0-30 d,the residue contents of MG in muscles increased with time,and then decreased gradually to 160. 00 μg/kg,achieving a dynamic balance. Meanwhile,the residue contents of LMG in muscles exhibited the similar changing manner compared with MG,reaching a dynamic balance with 3 370. 00 μg/kg. 3) in next year,the residue contents of malachite green in muscles from fresh cultured mud crap maintained between( 1. 43 ± 2. 45) μg/kg and( 4. 98 ± 2. 01) μg/kg during 0-70 d,and were undetected until 90 d. The residue contents of leucomalachite green increased from( 3. 10 ± 2. 31) μg/kg to( 105. 30 ± 8. 25) μg/kg at 0-5 d,and reached below the detection limit of 0. 50 μg/kg until 180 d in a fluctuant decreasing manner. The above results indicated that MG and LMG could be detected in the muscles of mud carp cultured in the positive ponds for two consecutive years,and the detection value in the first year was much higher than that in the second year. In the MG positive pond environment,MG and LMG were difficult to completely eliminate in a short period of time,and the residual enrichment could occur in the stocking of the muscle. This study could provide references for the supervision and law management administration to investigate the source of MG in aquatic products. It is of great significance to assess the quality and safety of aquatic products.
To study on enrichment effect of mud carp( Cirrhina mrigola) in malachite green( MG) positive environment and the elimination rules of malachite green in vivo,the mud carp with body weight of( 2. 68 ± 0. 51) g were fed in malachite green positive environment( ponds splashed with MG before) in the first year of June,and sampling at 0,0. 5,1,2,3,5,10,15,20 and 30 d after breeding. After changing breeding water in the next year of June,the mud crap,with similar body length and quality to previous samples,sampling at 0, 1, 2, 5, 10, 15, 20, 30, 40,50, 70,90, 120,150,180 and 210 d after feeding in the same pond. The residues of MG and LMG in fish muscles were determined by high performance liquid chromatography tandem mass spectrometry( HPLC-MS/MS) in order to evaluate the residual MG and LMG in stocking muscles in MG positive environment. The results showed that: 1) the established high performance liquid chromatography-tandem mass spectrometry method showed that MG and LMG had a good linear relationship in the range of 1. 0 ～ 200. 0 ng/m L respectively. The detection limits( LOD) and limit of quantitation( LOQ) of MG and LMG were both 0. 5 μg/kg. 2) during the first feeding 0-30 d,the residue contents of MG in muscles increased with time,and then decreased gradually to 160. 00 μg/kg,achieving a dynamic balance. Meanwhile,the residue contents of LMG in muscles exhibited the similar changing manner compared with MG,reaching a dynamic balance with 3 370. 00 μg/kg. 3) in next year,the residue contents of malachite green in muscles from fresh cultured mud crap maintained between( 1. 43 ± 2. 45) μg/kg and( 4. 98 ± 2. 01) μg/kg during 0-70 d,and were undetected until 90 d. The residue contents of leucomalachite green increased from( 3. 10 ± 2. 31) μg/kg to( 105. 30 ± 8. 25) μg/kg at 0-5 d,and reached below the detection limit of 0. 50 μg/kg until 180 d in a fluctuant decreasing manner. The above results indicated that MG and LMG could be detected in the muscles of mud carp cultured in the positive ponds for two consecutive years,and the detection value in the first year was much higher than that in the second year. In the MG positive pond environment,MG and LMG were difficult to completely eliminate in a short period of time,and the residual enrichment could occur in the stocking of the muscle. This study could provide references for the supervision and law management administration to investigate the source of MG in aquatic products. It is of great significance to assess the quality and safety of aquatic products.
引文
[1] Culp S J,Beland F A. Malachite green:a toxicological review[J]. J Am Coll Toxicol,1996,15(3):219-238.
[2]王声瑜.怎样有效选用含氯消毒剂和孔雀石绿[J].北京水产,1999(3):16.
[3] Plakas S M,Said K R,Stehly G R,et al. Uptake tissue distribution and metabolism of malachite green in the channel catfish(Ictalurus Punctatus)[J]. Can J Fish Aquat Scj,1996,53(6):1427-1433.
[4] Fernandes C,Lalitha V S,Rao V K. Enhancing effects of malachite green on development of hepatic preneoplastic lesions induced by N-nitrosodiethylamine in rats[J].Carcinogensis,1991,12(5):839-845.
[5] Rao K V K. Inhibition of DNA synthesis in primary rat hepatocyte cultures by malachite green:a new liver tumor promoter[J]. Toxicol Lett,1995,81(2/3):107-113.
[6]中华人民共和国农业部.中华人民共和国农业部第193号公告:食品动物禁用的兽药及其它化合物清单[EB/OL].(2011-04-22)[2018-01-05]. http://jiuban. moa. gov. cn/zwllm/tzgg/gg/201104/t20110422_1976324. htm.
[7]刘书贵,郑光明,尹怡,等.孔雀石绿及其代谢物在杂交鳢(斑鳢♀×乌鳢♂)体内的残留及消除规律研究[J].中国渔业质量与标准,2014,4(1):26-32.
[8]杨秋红,刘永涛,艾晓辉,等.孔雀石绿及其代谢物在斑点叉尾鮰体内及养殖环境中的消解规律[J].淡水渔业,2013,43(5):43-49.
[9]程杏安,郑子昭,蒋旭红,等.珠江三角洲水域底泥中孔雀石绿、结晶紫及其代谢物分析[J].生态科学,2015,34(3):59-64.
[10]陈培基,李刘冬,杨金兰,等.孔雀石绿在凡纳滨对虾体内的残留与消除规律[J].南方水产科学,2013,9(5):80-85.
[11]曲志娜,李存金,赵思俊,等.孔雀石绿及其代谢物在大菱鲆肌肉中的消除规律[J].中国农学通报,2008,24(6):491-496.
[12]黄金田,吕富,郑浩,等.孔雀石绿及其代谢物隐性孔雀石绿在中华绒螯蟹体中的消除规律[J].江苏农业科学,2010(2):326-328.
[13]高露姣,蔡友琼,姜朝军,等.孔雀石绿及其主要代谢物在欧洲鳗鲡肌肉中的蓄积及消除规律[J].水产学报,2007,31(9):104-108.
[14]邱绪建,林洪,王联珠,等.孔雀石绿及其代谢物无色孔雀石绿在鲫鱼肌肉中的代谢规律[J].中国海洋大学学报,2006,36(5):745-748.
[15]贾丽,王秀林,赵春晖,等.孔雀石绿及隐性孔雀石绿在罗非鱼体内残留与消除规律的研究[J].北京农业,2009(33):43-46.
[16]杨贤庆,孙满义,岑剑伟,等.孔雀石绿及其代谢产物在罗非鱼肌肉中残留规律的研究[J].热带海洋学报,2010,29(4):107-111.
[17]刘永涛,艾晓辉,索纹纹,等.浸泡条件下孔雀石绿及其代谢物隐色孔雀石绿在斑点叉尾鮰组织中分布及消除规律研究[J].水生生物学报,2013,37(2):269-280.
[18]李爱心.孔雀石绿在草鱼体内的代谢及残留研究[D].重庆:西南大学,2008.
[19] Zlatka B,Vlasta J,Ksenija S G. Elimination of malachite green residues from meat of rainbow trout and carp after water-born exposure[J]. Aquaculture,2011,321(1):13-16.
[20] Kirsi N,Tiina K,Erja L. Disappearance of malachite green residues in fry of rainbow trout Oncorhynchus mykiss after treatment of eggs at the hatching stage[J]. Aquaculture,2009,297:25-30.
[21]王志铮,施建军,吕敢堂.受短期饥饿胁迫下麦瑞加拉鲮鱼(Cirrhina mrigola)幼鱼的生长、肌体组分及其内脏消化酶活力的变化特征[J].海洋与湖沼,2006,37(3):218-224.
[22]国家质量监督检验检疫总局. GB/T19857—2005水产品中孔雀石绿和结晶紫残留量的测定[S].北京:中国标准出版社,2005.
[23]张彤晴,周刚,林海,等.孔雀石绿溶液水体自然降解初步研究[J].江苏农业科学,2007(1):211-214.
[2]王声瑜.怎样有效选用含氯消毒剂和孔雀石绿[J].北京水产,1999(3):16.
[3] Plakas S M,Said K R,Stehly G R,et al. Uptake tissue distribution and metabolism of malachite green in the channel catfish(Ictalurus Punctatus)[J]. Can J Fish Aquat Scj,1996,53(6):1427-1433.
[4] Fernandes C,Lalitha V S,Rao V K. Enhancing effects of malachite green on development of hepatic preneoplastic lesions induced by N-nitrosodiethylamine in rats[J].Carcinogensis,1991,12(5):839-845.
[5] Rao K V K. Inhibition of DNA synthesis in primary rat hepatocyte cultures by malachite green:a new liver tumor promoter[J]. Toxicol Lett,1995,81(2/3):107-113.
[6]中华人民共和国农业部.中华人民共和国农业部第193号公告:食品动物禁用的兽药及其它化合物清单[EB/OL].(2011-04-22)[2018-01-05]. http://jiuban. moa. gov. cn/zwllm/tzgg/gg/201104/t20110422_1976324. htm.
[7]刘书贵,郑光明,尹怡,等.孔雀石绿及其代谢物在杂交鳢(斑鳢♀×乌鳢♂)体内的残留及消除规律研究[J].中国渔业质量与标准,2014,4(1):26-32.
[8]杨秋红,刘永涛,艾晓辉,等.孔雀石绿及其代谢物在斑点叉尾鮰体内及养殖环境中的消解规律[J].淡水渔业,2013,43(5):43-49.
[9]程杏安,郑子昭,蒋旭红,等.珠江三角洲水域底泥中孔雀石绿、结晶紫及其代谢物分析[J].生态科学,2015,34(3):59-64.
[10]陈培基,李刘冬,杨金兰,等.孔雀石绿在凡纳滨对虾体内的残留与消除规律[J].南方水产科学,2013,9(5):80-85.
[11]曲志娜,李存金,赵思俊,等.孔雀石绿及其代谢物在大菱鲆肌肉中的消除规律[J].中国农学通报,2008,24(6):491-496.
[12]黄金田,吕富,郑浩,等.孔雀石绿及其代谢物隐性孔雀石绿在中华绒螯蟹体中的消除规律[J].江苏农业科学,2010(2):326-328.
[13]高露姣,蔡友琼,姜朝军,等.孔雀石绿及其主要代谢物在欧洲鳗鲡肌肉中的蓄积及消除规律[J].水产学报,2007,31(9):104-108.
[14]邱绪建,林洪,王联珠,等.孔雀石绿及其代谢物无色孔雀石绿在鲫鱼肌肉中的代谢规律[J].中国海洋大学学报,2006,36(5):745-748.
[15]贾丽,王秀林,赵春晖,等.孔雀石绿及隐性孔雀石绿在罗非鱼体内残留与消除规律的研究[J].北京农业,2009(33):43-46.
[16]杨贤庆,孙满义,岑剑伟,等.孔雀石绿及其代谢产物在罗非鱼肌肉中残留规律的研究[J].热带海洋学报,2010,29(4):107-111.
[17]刘永涛,艾晓辉,索纹纹,等.浸泡条件下孔雀石绿及其代谢物隐色孔雀石绿在斑点叉尾鮰组织中分布及消除规律研究[J].水生生物学报,2013,37(2):269-280.
[18]李爱心.孔雀石绿在草鱼体内的代谢及残留研究[D].重庆:西南大学,2008.
[19] Zlatka B,Vlasta J,Ksenija S G. Elimination of malachite green residues from meat of rainbow trout and carp after water-born exposure[J]. Aquaculture,2011,321(1):13-16.
[20] Kirsi N,Tiina K,Erja L. Disappearance of malachite green residues in fry of rainbow trout Oncorhynchus mykiss after treatment of eggs at the hatching stage[J]. Aquaculture,2009,297:25-30.
[21]王志铮,施建军,吕敢堂.受短期饥饿胁迫下麦瑞加拉鲮鱼(Cirrhina mrigola)幼鱼的生长、肌体组分及其内脏消化酶活力的变化特征[J].海洋与湖沼,2006,37(3):218-224.
[22]国家质量监督检验检疫总局. GB/T19857—2005水产品中孔雀石绿和结晶紫残留量的测定[S].北京:中国标准出版社,2005.
[23]张彤晴,周刚,林海,等.孔雀石绿溶液水体自然降解初步研究[J].江苏农业科学,2007(1):211-214.