The persistence of malachite green (MG), and its metabolite leucomalachite green (LMG), in fish tissues is still unclear, leading to many trade disputes. In this research, we established and evaluated an HPLC method t...The persistence of malachite green (MG), and its metabolite leucomalachite green (LMG), in fish tissues is still unclear, leading to many trade disputes. In this research, we established and evaluated an HPLC method that could detect MG and LMG simultaneously, and then investigated the persistence of these two toxins in the tissues of juvenile perch (Lateolabrax japonicus) post sub-chronic MG exposure at 1.0 mg/L. Exposure lasted for 2 h everyday and was repeated six times. The perch were then placed in MG-free seawater for 100 d to eliminate the toxins. Results show that MG accumulated in the tissues, including the gills, liver, muscle, blood and viscera, and then was metabolized rapidly to LMG. The concentrations of these two toxins increased significantly with the accumulation process. In general, the highest concentrations of MG and LMG in all tissue exceeded 1 000 μg/kg, except for MG in the muscle. The order of accumulation levels (highest to lowest) of MG was gill>blood>liver>viscera>muscle, while that of LMG was liver>blood>gill>viscera>muscle. High levels of MG or LMG could persist for several hours but decreased rapidly during the elimination process. The concentration of LMG was much higher than that of MG during the experiment, especially in the gill, liver and blood. Therefore, the three tissues play important roles in toxin accumulation, biotransformation, and elimination. Although the MG and LMG concentrations in muscle were much lower than in other tissues, the content still exceeded the European minimum required performance limit (MRPL), even after 2 400 h (100 d) of elimination. This demonstrates that it is extremely difficult to eliminate MG and LMG from tissues of perch, and therefore use of these toxins is of concern to public health.展开更多
Polymerized high internal phase emulsion(polyHIPE)monoliths were prepared and applied as adsorbent materials for solid-phase extraction(SPE)of malachite green(MG)and leucomalachite green(LMG)from water samples.The pol...Polymerized high internal phase emulsion(polyHIPE)monoliths were prepared and applied as adsorbent materials for solid-phase extraction(SPE)of malachite green(MG)and leucomalachite green(LMG)from water samples.The polyHIPE monoliths were prepared by post-functionalization of monolithic surface with 6-aminocaproic acid(ACA)via ring opening reaction of epoxy groups in glycidyl methacrylate(GMA)-based polyHIPEs,and then applied to the preconcentration and determination of trace MG and LMG in environmental water samples by combing with high-performance liquid chromatog-raphy(HPLC).Taking MG and LMG as targets,main factors affecting SPE performance of the polyHIPE monoliths were investigated.Under the optimized conditions,the ACA-functionalized polyHIPE monoliths could effectively preconcentrate MG and LMG from 150 mL of water samples,and the recoveries of MG and LMG at three spiked levels were ranged from 84.8 to 97.4%with the relative standard deviations(RSDs)lower than 6%.The proposed method exhibited good linearity in the range of 2-200 ng mL^(-1),with low limits of detection of 17.0 and 8.7 pg mL^(-1)for MG and LMG,respectively.In addi-tion,the prepared ACA-modified polyHIPE monolith showed good durability and stability,and it could be reused for 200 cycles without obvious losing the extraction performance.展开更多
建立了水产品中孔雀石绿(MG)和结晶紫(CV)及其代谢物隐色孔雀石绿(LMG)和隐色结晶紫(LCV)残留的QuEChERS/UPLC-MS/MS分析方法。样品采用乙腈提取,改进的QuEChERS(EMR-Lipid)分散固相萃取净化,经Agilent Eclipse Plus C_(18)(1.8μm,3.0 ...建立了水产品中孔雀石绿(MG)和结晶紫(CV)及其代谢物隐色孔雀石绿(LMG)和隐色结晶紫(LCV)残留的QuEChERS/UPLC-MS/MS分析方法。样品采用乙腈提取,改进的QuEChERS(EMR-Lipid)分散固相萃取净化,经Agilent Eclipse Plus C_(18)(1.8μm,3.0 mm×100 mm)色谱柱分离,电喷雾串联四极杆质谱多反应监测正离子方式测定。4种分析物在0.2~10.0μg/L范围内线性关系良好,相关系数均大于0.997。鱼肉中4种分析物在0.5,1.0,5.0μg/kg加标浓度水平下,回收率为77.1%~106.6%,相对标准偏差(RSD)为1.3%~4.3%。该方法简单、稳定、可靠,能有效去除样品中的蛋白质、脂肪等大分子杂质,可满足水产品中孔雀石绿、结晶紫以及隐色代谢物残留检测与确证的需要。展开更多
A high performance liquid chromatography method had been established for the determination of malachite green and leucomalachite green in aquatic water.Malachite green and leucomalachite green were extracted with dich...A high performance liquid chromatography method had been established for the determination of malachite green and leucomalachite green in aquatic water.Malachite green and leucomalachite green were extracted with dichloromethane,then concentrated,and dissolved with acetonitrile.Samples were chromatographed using an acetonitrile-ammonium acetate buffer mobile phase on a C18 column in line with a post column reactor filled with lead(IV) oxide and celite.The linear range was from 0.01 μg·mL-1 to 1 μg·mL1 with r2 of 0.999 9.The average recoveries of MG were 83.00%,86.40%,93.50%,and LMG were 74.00%,89.00%,91.80% when samples were spiked with MG and LMG of 0.1,0.5,1.0 μg·mL-1 respectively.The detection limits of MG and LMG were 0.003 μg·mL-1 and 0.004 μg·mL-1 respectively.The RSDs were 4.278%-6.250% and 2.179%-6.757% respectively.The method is sensitive,accurate and suitable for the determination of trace malachite green and leucomalachite green in aquatic water.展开更多
以7 mg/L的孔雀石绿浸泡斑点叉尾苗种5min后将其饲养于池塘的网箱中,研究了在养殖模式下孔雀石绿及其代谢物隐色孔雀石绿在斑点叉尾苗种各组织中的分布及消除规律。采用高效液相色谱串联质谱法(HPLC-MS/MS)分析孔雀石绿及其代谢物...以7 mg/L的孔雀石绿浸泡斑点叉尾苗种5min后将其饲养于池塘的网箱中,研究了在养殖模式下孔雀石绿及其代谢物隐色孔雀石绿在斑点叉尾苗种各组织中的分布及消除规律。采用高效液相色谱串联质谱法(HPLC-MS/MS)分析孔雀石绿及其代谢物隐色孔雀石绿在斑点叉尾血液、肌肉、皮肤、肝脏、肾脏组织中的浓度水平。采用药代动力学分析软件3p97对血药浓度时间数据进行分析。结果表明,孔雀石绿和隐色孔雀石绿血药浓度时间曲线符合有吸收二室模型,动力学方程分别为:C孔雀石绿=683.063 e 0.248 t+11.176e 0.006 t 694.239e 0.333 t,C隐色孔雀石绿=757.240 e 0.222 t+14.474 e 0.007 t–771.714 e 0.382 t。血液中孔雀石绿和隐色孔雀石绿达峰时间Tpeak分别为3.480和3.623h,峰浓度值Cmax分别为81.560和159.619 ng/mL,表观分布容积Vd/F分别为37.689和21.125 L/kg,分布相的一级速率常数α分别为0.248和0.222/h,消除相的一级速率常数β分别为0.006和0.007/h,吸收半衰期T(1/2)α分别为2.794和3.124h,消除半衰期T(1/2)β分别为113.068和105.841h,中央室向周边室转运的一级速率常数K12分别为0.020和0.015/h,周边室向中央室转运的一级速率常数K21分别为0.159和0.121/h,药-时曲线下面积AUC分别为2493.944和3601.863 ng.h/mL。肌肉、皮肤、肝脏和肾脏组织中孔雀石绿和隐色孔雀石绿浓度水平的结果表明,孔雀石绿在斑点叉尾4种组织中浓度由高到低的顺序是皮肤>肌肉>肾脏>肝脏,其中斑点叉尾皮肤组织易蓄积孔雀石绿,其残留时间最长,肝脏组织由于对孔雀石绿有极强的代谢转化功能而浓度较低。孔雀石绿在肌肉、皮肤、肝脏和肾脏组织中的消除方程分别为C=5.570 e 0.009t、C=6.302 e 0.007t、C=4.791 e 0.006t和C=4.591 e 0.002t,相关系数r2≥0.773,消除半衰期T1/2肌肉、皮肤、肝脏和肾脏分别为3.2、4.1、4.8和14.4d。肌肉、皮肤、肝脏和肾脏组织中孔雀石绿分别在45、60、30和60d才未被检测到;隐色孔雀石绿在斑点叉尾4种组织中浓度由高到低的顺序是肝脏>皮肤>肌肉>肾脏,残留时间最长的组织也是皮肤组织。隐色孔雀石绿在肌肉、皮肤、肝脏和肾脏组织中的消除方程分别为C=6.491 e 0.004t、C=6.958 e 0.003t、C=6.722 e 0.007t和C=6.162 e 0.002t,相关系数r2≥0.673,消除半衰期T1/2肌肉、皮肤、肝脏和肾脏分别为7.2、9.6、4.1和14.4d。肌肉、皮肤、肝脏和肾脏组织中隐色孔雀石绿分别在90、90、60和90d才未被检出。试验期间(2011年5月17日至7月15日)平均水温为26.4℃,孔雀石绿和隐色孔雀石绿90d后在各组织中才未检测到,因此,使用7 mg/L孔雀石绿浸泡2龄斑点叉尾苗种孔雀石绿及其代谢物隐色孔雀石绿至少应经过2376℃.d后才能消除。展开更多
基金Supported by the Special Fund of Chinese Government for Basic Scientific Research Operations in Commonweal Research Institute (Yellow Sea Fisheries Research Institute) (No. 2007-qn-12)the Strategic Research Grant of the Databases and Risk Analysis of POPs in Aquatic Products (No. 2005DIB4J049)the Standard System Research on Quality and Safety of Aquatic Products (No. 2004DEA70880)
文摘The persistence of malachite green (MG), and its metabolite leucomalachite green (LMG), in fish tissues is still unclear, leading to many trade disputes. In this research, we established and evaluated an HPLC method that could detect MG and LMG simultaneously, and then investigated the persistence of these two toxins in the tissues of juvenile perch (Lateolabrax japonicus) post sub-chronic MG exposure at 1.0 mg/L. Exposure lasted for 2 h everyday and was repeated six times. The perch were then placed in MG-free seawater for 100 d to eliminate the toxins. Results show that MG accumulated in the tissues, including the gills, liver, muscle, blood and viscera, and then was metabolized rapidly to LMG. The concentrations of these two toxins increased significantly with the accumulation process. In general, the highest concentrations of MG and LMG in all tissue exceeded 1 000 μg/kg, except for MG in the muscle. The order of accumulation levels (highest to lowest) of MG was gill>blood>liver>viscera>muscle, while that of LMG was liver>blood>gill>viscera>muscle. High levels of MG or LMG could persist for several hours but decreased rapidly during the elimination process. The concentration of LMG was much higher than that of MG during the experiment, especially in the gill, liver and blood. Therefore, the three tissues play important roles in toxin accumulation, biotransformation, and elimination. Although the MG and LMG concentrations in muscle were much lower than in other tissues, the content still exceeded the European minimum required performance limit (MRPL), even after 2 400 h (100 d) of elimination. This demonstrates that it is extremely difficult to eliminate MG and LMG from tissues of perch, and therefore use of these toxins is of concern to public health.
基金supported by the National Natural Science Foundation of China(Grant No.21964006,and 21665006)the Natural Science Foundation from Hunan Province(No.2020JJ4640 and 2017JJ3342)+1 种基金the Natural Science Foundation from Guangxi Zhuang Autonomous Region(No.2018GXNSFAA138134)the Scientific Research Found of Changsha University(SF1934),respectively.
文摘Polymerized high internal phase emulsion(polyHIPE)monoliths were prepared and applied as adsorbent materials for solid-phase extraction(SPE)of malachite green(MG)and leucomalachite green(LMG)from water samples.The polyHIPE monoliths were prepared by post-functionalization of monolithic surface with 6-aminocaproic acid(ACA)via ring opening reaction of epoxy groups in glycidyl methacrylate(GMA)-based polyHIPEs,and then applied to the preconcentration and determination of trace MG and LMG in environmental water samples by combing with high-performance liquid chromatog-raphy(HPLC).Taking MG and LMG as targets,main factors affecting SPE performance of the polyHIPE monoliths were investigated.Under the optimized conditions,the ACA-functionalized polyHIPE monoliths could effectively preconcentrate MG and LMG from 150 mL of water samples,and the recoveries of MG and LMG at three spiked levels were ranged from 84.8 to 97.4%with the relative standard deviations(RSDs)lower than 6%.The proposed method exhibited good linearity in the range of 2-200 ng mL^(-1),with low limits of detection of 17.0 and 8.7 pg mL^(-1)for MG and LMG,respectively.In addi-tion,the prepared ACA-modified polyHIPE monolith showed good durability and stability,and it could be reused for 200 cycles without obvious losing the extraction performance.
文摘A high performance liquid chromatography method had been established for the determination of malachite green and leucomalachite green in aquatic water.Malachite green and leucomalachite green were extracted with dichloromethane,then concentrated,and dissolved with acetonitrile.Samples were chromatographed using an acetonitrile-ammonium acetate buffer mobile phase on a C18 column in line with a post column reactor filled with lead(IV) oxide and celite.The linear range was from 0.01 μg·mL-1 to 1 μg·mL1 with r2 of 0.999 9.The average recoveries of MG were 83.00%,86.40%,93.50%,and LMG were 74.00%,89.00%,91.80% when samples were spiked with MG and LMG of 0.1,0.5,1.0 μg·mL-1 respectively.The detection limits of MG and LMG were 0.003 μg·mL-1 and 0.004 μg·mL-1 respectively.The RSDs were 4.278%-6.250% and 2.179%-6.757% respectively.The method is sensitive,accurate and suitable for the determination of trace malachite green and leucomalachite green in aquatic water.
文摘以7 mg/L的孔雀石绿浸泡斑点叉尾苗种5min后将其饲养于池塘的网箱中,研究了在养殖模式下孔雀石绿及其代谢物隐色孔雀石绿在斑点叉尾苗种各组织中的分布及消除规律。采用高效液相色谱串联质谱法(HPLC-MS/MS)分析孔雀石绿及其代谢物隐色孔雀石绿在斑点叉尾血液、肌肉、皮肤、肝脏、肾脏组织中的浓度水平。采用药代动力学分析软件3p97对血药浓度时间数据进行分析。结果表明,孔雀石绿和隐色孔雀石绿血药浓度时间曲线符合有吸收二室模型,动力学方程分别为:C孔雀石绿=683.063 e 0.248 t+11.176e 0.006 t 694.239e 0.333 t,C隐色孔雀石绿=757.240 e 0.222 t+14.474 e 0.007 t–771.714 e 0.382 t。血液中孔雀石绿和隐色孔雀石绿达峰时间Tpeak分别为3.480和3.623h,峰浓度值Cmax分别为81.560和159.619 ng/mL,表观分布容积Vd/F分别为37.689和21.125 L/kg,分布相的一级速率常数α分别为0.248和0.222/h,消除相的一级速率常数β分别为0.006和0.007/h,吸收半衰期T(1/2)α分别为2.794和3.124h,消除半衰期T(1/2)β分别为113.068和105.841h,中央室向周边室转运的一级速率常数K12分别为0.020和0.015/h,周边室向中央室转运的一级速率常数K21分别为0.159和0.121/h,药-时曲线下面积AUC分别为2493.944和3601.863 ng.h/mL。肌肉、皮肤、肝脏和肾脏组织中孔雀石绿和隐色孔雀石绿浓度水平的结果表明,孔雀石绿在斑点叉尾4种组织中浓度由高到低的顺序是皮肤>肌肉>肾脏>肝脏,其中斑点叉尾皮肤组织易蓄积孔雀石绿,其残留时间最长,肝脏组织由于对孔雀石绿有极强的代谢转化功能而浓度较低。孔雀石绿在肌肉、皮肤、肝脏和肾脏组织中的消除方程分别为C=5.570 e 0.009t、C=6.302 e 0.007t、C=4.791 e 0.006t和C=4.591 e 0.002t,相关系数r2≥0.773,消除半衰期T1/2肌肉、皮肤、肝脏和肾脏分别为3.2、4.1、4.8和14.4d。肌肉、皮肤、肝脏和肾脏组织中孔雀石绿分别在45、60、30和60d才未被检测到;隐色孔雀石绿在斑点叉尾4种组织中浓度由高到低的顺序是肝脏>皮肤>肌肉>肾脏,残留时间最长的组织也是皮肤组织。隐色孔雀石绿在肌肉、皮肤、肝脏和肾脏组织中的消除方程分别为C=6.491 e 0.004t、C=6.958 e 0.003t、C=6.722 e 0.007t和C=6.162 e 0.002t,相关系数r2≥0.673,消除半衰期T1/2肌肉、皮肤、肝脏和肾脏分别为7.2、9.6、4.1和14.4d。肌肉、皮肤、肝脏和肾脏组织中隐色孔雀石绿分别在90、90、60和90d才未被检出。试验期间(2011年5月17日至7月15日)平均水温为26.4℃,孔雀石绿和隐色孔雀石绿90d后在各组织中才未检测到,因此,使用7 mg/L孔雀石绿浸泡2龄斑点叉尾苗种孔雀石绿及其代谢物隐色孔雀石绿至少应经过2376℃.d后才能消除。
