孔雀石绿和结晶紫检测新方法研究
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摘要
目的:孔雀石绿和结晶紫等潜在致癌物,是近年来水体污染和食用水产品卫生安全中重点监控的有机污染物。目前,孔雀石绿和结晶紫的主要检测方法是高效液相色谱法和高效液相色谱-质谱法。共振瑞利散射(RRS)光谱是近年发展起来的一项高灵敏度定量分析新技术,在生命科学、环境科学和纳米材料研究等方面得到了广泛的应用,但未见用RRS测定孔雀石绿和结晶紫的报道。本文利用共振瑞利散射法的技术特点,建立灵敏度高、选择性好、操作简单的检测孔雀石绿和结晶紫的新方法。
方法:孔雀石绿和结晶紫都是三苯甲烷类阳离子染料,在一定条件下,将其与大阴离子反应,形成离子缔合物而失去亲水性。离子缔合物能在水相中因疏水作用而相互聚集,并以纳米粒子保留在溶液中。本文采用负吸收效应,从理论上对体系基体的作用进行分析;采用紫外-可见分光光度计测定吸收光强度,求出缔合物的条件稳定常数,对体系酸效应进行分析;采用荧光分光光度计测定共振瑞利散射强度,对孔雀石绿和结晶紫进行定性、定量分析。
结果:本文以负吸收效应和条件稳定常数分别分析了基体和酸度对共振瑞利散射体系的影响;以I_3~-阴离子和磷钼酸根阴离子为配体,建立了共振瑞利散射测定孔雀石绿的两种新方法;以I3-阴离子为配体,建立了共振瑞利散射法测定结晶紫的新方法。I_3~--孔雀石绿体系中,孔雀石绿的检出限为3.6ng/mL,线性范围:12～900ng/mL,相关系数r=0.9997;磷钼酸根-孔雀石绿体系中,孔雀石绿的检出限为55ng/mL,线性范围:0.18～5.0μg/mL,相关系数r=0.9998。I_3~--结晶紫体系中,结晶紫的检出限为0.009μg/mL,线性范围:0.03～1.80μg/mL,相关系数r=0.9997。
结论:利用共振瑞利散射技术建立水产品和水样中孔雀石绿和结晶检测的新方法,灵敏度达到或接近高效液相色谱法水平,但与高效液相色谱法和高效液相色谱-质谱法相比,新方法具有仪器试剂易得、操作简单、分析时间短、使用无二次污染等优点,更便于在基层检测部门推广应用。
Objective:
Some potential carcinogens, such as malachite green (MG) and crystal violet (CV), are the significant organic contaminants, which are monitored for the safety of food or water. At present, the main detection methods of MG and CV are liquid chromatography and liquid chromatography tandem mass spectrometry. Resonance Rayleigh scattering (RRS) spectrum is a new sensitive quantitative analytical technique, which has been steadily developed recently, and has been applied widely to the study of life, environmental sciences and nanometer materials, etc. However, there are no reports discussing the determination of MG and CV by RRS. In this work, a sensitive, selective, simple and rapid method based on RRS was established for the determination of MG and CV.
Methods:
Triphenylmethane dyes MG and CV respectively react with some negative ions under suitable conditions to form a hydrophobic ion-associate. Then the ion-associate could self-assemble to form a nanoparticle, suspending in aqueous solution at suitable concentration and emitting intense RRS induced by excited ray. In this article, the effect of matrix on the intensity of RRS is analyzed theoretically based on the matrix negative absorption effect; the effect of pH is analyzed based on the conditional stability constant which is assayed by spectrophotometry; and MG and CV are analyzed qualitatively and quantitatively through detecting the intensity of RRS by the fluorescence spectrophotometer synchronously scanning.
Results:
In this article, the effects of matrix and pH on the intensity of RRS were analyzed based on the matrix negative absorption effect and conditional stability constant for the first time; novel methods were developed to analyze MG by RRS based on the forming of ion-associate of MG-I3- and MG-PMA (phosphomolybdate) respectively, and similarly a novel method was developed to analyze CV through the forming of ion-associate of CV-I_3~-. In the system of MG-I_3~-, the detection limit of the method is 3.6 ng/mL, the linearity range is 12～900 ng/mL, and the relative index r is 0.9997. In the system of MG-PMA, the detection limit of the method is 0.055μg/mL, the linearity range is 0.18～5.00μg/mL, and the relative index r is 0.9998. And In the system of CV-I_3~-, the detection limit of the method is 0.009μg/mL, the linearity range is 0.03～1.80μg/mL, and the relative index r is 0.9997.
Conclusion:
The sensitivity of the new method is similar to liquid chromatography and liquid chromatography tandem mass spectrometry, but RRS is simpler, faster and has no secondary pollution. So it is more suitable for the determination of MG and CV in the lower detection units.
方法:孔雀石绿和结晶紫都是三苯甲烷类阳离子染料,在一定条件下,将其与大阴离子反应,形成离子缔合物而失去亲水性。离子缔合物能在水相中因疏水作用而相互聚集,并以纳米粒子保留在溶液中。本文采用负吸收效应,从理论上对体系基体的作用进行分析;采用紫外-可见分光光度计测定吸收光强度,求出缔合物的条件稳定常数,对体系酸效应进行分析;采用荧光分光光度计测定共振瑞利散射强度,对孔雀石绿和结晶紫进行定性、定量分析。
结果:本文以负吸收效应和条件稳定常数分别分析了基体和酸度对共振瑞利散射体系的影响;以I_3~-阴离子和磷钼酸根阴离子为配体,建立了共振瑞利散射测定孔雀石绿的两种新方法;以I3-阴离子为配体,建立了共振瑞利散射法测定结晶紫的新方法。I_3~--孔雀石绿体系中,孔雀石绿的检出限为3.6ng/mL,线性范围:12～900ng/mL,相关系数r=0.9997;磷钼酸根-孔雀石绿体系中,孔雀石绿的检出限为55ng/mL,线性范围:0.18～5.0μg/mL,相关系数r=0.9998。I_3~--结晶紫体系中,结晶紫的检出限为0.009μg/mL,线性范围:0.03～1.80μg/mL,相关系数r=0.9997。
结论:利用共振瑞利散射技术建立水产品和水样中孔雀石绿和结晶检测的新方法,灵敏度达到或接近高效液相色谱法水平,但与高效液相色谱法和高效液相色谱-质谱法相比,新方法具有仪器试剂易得、操作简单、分析时间短、使用无二次污染等优点,更便于在基层检测部门推广应用。
Objective:
Some potential carcinogens, such as malachite green (MG) and crystal violet (CV), are the significant organic contaminants, which are monitored for the safety of food or water. At present, the main detection methods of MG and CV are liquid chromatography and liquid chromatography tandem mass spectrometry. Resonance Rayleigh scattering (RRS) spectrum is a new sensitive quantitative analytical technique, which has been steadily developed recently, and has been applied widely to the study of life, environmental sciences and nanometer materials, etc. However, there are no reports discussing the determination of MG and CV by RRS. In this work, a sensitive, selective, simple and rapid method based on RRS was established for the determination of MG and CV.
Methods:
Triphenylmethane dyes MG and CV respectively react with some negative ions under suitable conditions to form a hydrophobic ion-associate. Then the ion-associate could self-assemble to form a nanoparticle, suspending in aqueous solution at suitable concentration and emitting intense RRS induced by excited ray. In this article, the effect of matrix on the intensity of RRS is analyzed theoretically based on the matrix negative absorption effect; the effect of pH is analyzed based on the conditional stability constant which is assayed by spectrophotometry; and MG and CV are analyzed qualitatively and quantitatively through detecting the intensity of RRS by the fluorescence spectrophotometer synchronously scanning.
Results:
In this article, the effects of matrix and pH on the intensity of RRS were analyzed based on the matrix negative absorption effect and conditional stability constant for the first time; novel methods were developed to analyze MG by RRS based on the forming of ion-associate of MG-I3- and MG-PMA (phosphomolybdate) respectively, and similarly a novel method was developed to analyze CV through the forming of ion-associate of CV-I_3~-. In the system of MG-I_3~-, the detection limit of the method is 3.6 ng/mL, the linearity range is 12～900 ng/mL, and the relative index r is 0.9997. In the system of MG-PMA, the detection limit of the method is 0.055μg/mL, the linearity range is 0.18～5.00μg/mL, and the relative index r is 0.9998. And In the system of CV-I_3~-, the detection limit of the method is 0.009μg/mL, the linearity range is 0.03～1.80μg/mL, and the relative index r is 0.9997.
Conclusion:
The sensitivity of the new method is similar to liquid chromatography and liquid chromatography tandem mass spectrometry, but RRS is simpler, faster and has no secondary pollution. So it is more suitable for the determination of MG and CV in the lower detection units.
引文
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[7] Srivastava S.J, Singh N.D, Srivastava A.K, et al. Acute toxicity of malachite green and its effects on certain blood parameters of a catfish, Heteropneustes fossilis[J]. Aquat. Toxicol, 1995, 31: 241～247
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[11] Srivastava A.K, Sinha R, Singh N.D, et al. Histopathological changes in a freshwater catfish, Heteropneustes fossilis following exposure to malachitegreen[J]. Proc. Natl. Acad. Sci. India, 1998, 68 (B): 123～127
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[16] Steven M Plakas. Optimization of a liquid chromatographic method for determination of malachite green and its metabolites in fish tissues[J]. J of AOAC, 1995, 78: 1388～1394
[17] Srivastava A.K, Sinha R, Singh N.D, et al. Malachite green induced changes in carbohydrate metabolism and blood chloride levels in the freshwater catfish, Heteropneustes fossilis[J]. Acta Hydrobiol, 1995, 37(2): 113～119
[18] Yildiz H.Y, Pulatsu S. Evaluation of the secondary stress response in healthy Nile tilapia (Oreochromis niloticus L.) after treatment with a mixture of formaline, malachite green and methylene blue[J]. Aquacult Res, 1999, 30(5): 379～383
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[20] Doerge D.R, Chang H.C, Divi R.L, et al. Mechanism for inhibition of thyroid peroxidase by leucomalahchite green[J]. Chem. Res. Toxicol, 1998, 11(9): 1098～1104
[21] Tanck M.W.T, Hajee C.A.J, Olling M, et al. Negative effect of malachite green on haematocrit of rainbow trout(Oncorhynchus mykiss Walbaum)[J]. Bull. Eur. Assoc. Fish. Pathol, 1995, 15(4): 134～136
[22] Srivastava A.K, Roy D, Sinha R, et al. Dyes induced changes in the haematological parameters of a freshwater catfish, Heteropneutes fossilis[J]. Ecol. Environ. Conserv, 1996, 2: 155～158
[23] Grizzle, J. M. Hematological changes in fingerling channel catfish exposed to malachite green. Prog[J]. Fish Cult, 1977, 39 (2): 90～93
[24] 翟毓秀, 郭莹莹, 耿霞. 孔雀石绿的代谢机理及生物毒性研究进展[J]. 中国海洋大学学报, 2007, 37 (1): 27～32
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[28] Culp S. J. Metabolic changes occurring in mice and rats fed LMG[C]. San Diego: Proc 88th Ann Meeting of Am Soc for Cancer Re2 search, 1997: 121
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[30] 李宁. 什么是孔雀石绿. 知识卡片, 2005, 11: 79
[31] Larry G. Rushing, Harold C. Thompson. Simultaneous determination of malachite green, gentian violet and their leuco metabolites in catfish or trouttissue by high performance liquid chromatography with visible detection[J]. Journal of Chromatography B, 1997, 688: 325～330
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