四种人工纳米颗粒物在水相中的转化行为及生态毒理效应
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摘要
伴随着各种纳米材料的广泛应用,人工纳米颗粒物(MNPs)将不可避免地释放到环境中,成为潜在的污染物,对生态健康和人体健康产生潜在不利影响。释放到环境中的MNPs可发生多种迁移和转化过程,进而影响其生态毒理学效应。如何评价和预测MNPs的水环境行为(尤其是MNPs与溶解性有机质(DOM)的相互作用及其对MNPs胶体团聚状态的影响)及对水生生物的毒理效应,是纳米生态毒理学研究的前沿领域。本文基于分子模拟方法,研究DOM与富勒烯(C60)的相互作用机制,阐明了DOM对C60在水中溶解与团聚行为的影响机制；通过生物测试,考察了纳米二氧化钛(nTiO2).纳米二氧化铈(nCeO2)以及纳米银(nAg)颗粒物对不同营养级水生生物的毒理效应,并探讨了其毒性作用机制。主要研究内容和研究成果如下：
(1)利用分子力学和量子化学计算,模拟了C60与不同来源的DOM模型分子间的相互作用。相互作用能的计算结果表明,DOM模型分子可与C60间形成稳定的复合物。与来源于淡水或褐煤的DOM模型分子相比较,土壤中的DOM模型分子与C60间形成的复合物的稳定性更高。电荷转移分析的结果表明,DOM在C60-DOM的复合物中是电子受体。此外,DOM的存在显著地提高了C60的表观水溶解度(SC60/DOM),且SC60/DOM随C60-DOM复合物的前线分子轨道能级差的增加而降低。
(2)采用荧光探针法研究了nTiO2和nCeO2胶体对斜生栅藻(Scenedesmus obliquus)细胞膜通透性和膜电位的影响。利用DLVO理论对这两种MNPs的胶体稳定性进行了评价。在总势能曲线上nTiO2胶体的势能峰值是nCeO2的4.2倍,表明nTiO2胶体的稳定性高于nCeO2胶体。nTiO2胶体降低了绿藻细胞膜通透性,nCeO2胶体却增加了绿藻细胞膜通透性。这两种MNPs胶体均引起细胞膜电位的升高,且nTiO2胶体诱导细胞膜电位升高程度明显高于nCeO2胶体,表明干扰绿藻细胞膜的正常生理功能是MNPs对绿藻的毒性作用机制之一。与nCeO2胶体相比,具有更高悬浮稳定性的nTiO2胶体表现出更强的绿藻细胞膜毒性效应,说明MNPs的胶体稳定性是决定其毒性差异的重要因素。
(3)考察了nAg胶体对三种不同营养级水生生物的毒理效应,包括羊角月牙藻(Raphidocelis subcapitata)、圆形盘肠溞(Chydorus sphaericus)及斑马鱼(Danio rerio)胚胎。通过反应加和模型定量评价了游离态银离子(Ag+)对nAg胶体的毒性贡献。研究了三种商业nAg颗粒物：无包覆剂(Bare)nAg、聚乙烯吡咯烷酮包覆(PVP)nAg、悬浮剂分散(DIS)nAg。发现这三种nAg胶体对受试生物的毒性(以半数效应浓度为基础)大小顺序均为：DIS-nAg> PVP-nAg> Bare-nAg。DIS-nAg含有最高浓度的游离态Ag+,而Bare-nAg中的游离态Ag+的浓度最低,表明主解释nAg的毒性机制时不能忽略游离态Ag+的作用。此外,游离态Ag+对nAg胶体的Raphidocelis subcapitata毒性贡献最大,而对nAg胶体的Danio rerio毒性贡献最小,说国游离态Ag+对nAg颗粒对不同水生生物的毒性贡献不同,取决于水生生物的营养级水平。
(4)研究了DOM对nAg胶体水生毒性的影响。以一种腐殖酸模型分子(HS)来模拟DOM,考察了HS对PVP包覆的nAg(一次粒径为20nm)对羊角月牙藻、圆形盘肠溞及斑马鱼胚胎的毒性影响。发现随HS浓度的升高,nAg胶体对三种水生生物的毒性降低。随着HS浓度的增加,nAg胶体颗粒物的尺度更小,意味颗粒物的尺度变化不是导致HS存在条件下nAg毒性降低的原因。HS浓度增高时,导致nAg表面电荷更负,说明表面电荷大小也不是导致有HS存在时nAg毒性降低的决定因素；HS浓度较高时,Ag+释放受到了抑制,以nAg胶体中游离态Ag+浓度来表达胶体的毒性反应时,存在明显的浓度-反应关系,说明HS通过抑制Ag+的释放来降低nAg胶体的水生毒性。
With the increasing use of various nanostructured materials, manufacuted nanoparticles (MNPs) are inevitablily discharged into the environment. Thereby MNPs could become potential pollutants, and pose a potential health risk to human and ecological species. Once released into the environment, MNPs can undergo many transport and transformation processes affecting their ecotoxicity. How to assess and predict aquantic behavior (particular for the interaction of MNPs with dissolved organic matter (DOM) and their effects on agglomeration of MNPs colloids) and toxicological effects to aquatic organisms is the frontier research in the field of nanoecotoxicology. In this thesis, interactions of fullerene (C6o) with dissolved organic matters (DOM) were investigated by the means of molecular modelling, and the mechanism by which DOM influence dissolution and agglomeration behavior of C6o was elucidated. In addition, toxicological effects of nano-titanium dioxide (nTiO2), nano-cerium dioxide (nCeO2) and nano-silver (nAg) colloids on aquatic organisms were observed and mechanisms of action for their aquatic toxicity were illustrated. Main contents and results are as follows:
(1) The interaction of fullerene C60with DOM analogues is computationally simulated by molecular mechanics and density functional theory. The calculated interaction energies displayed that the DOM could stablize C60, and the stability between C60and the soil DOM analogues is greater than that between C60and the DOM analogues from fresh water or coal. The computed electrostatic potential indicates that DOM analogues are electron acceptors in the C60-DOM complexes. The presence of DOM increases the apparent water solubility of C60-It is also observed that the C60apparent water solubility decreases with the increase of the energy gaps of frontier molecular orbitals (ELUMO-EHOMO) for each C60-DOM complex.
(2) The effects of nTiO2and nCeO2colloids on cell membrane permeability and cell membrane potential of a green alga (Scenedesmus obliquus) were investigated by the fluorescent probe technique. The stability of nanoparticle colloids was evaluated by the DLVO theory. The results show that the magnitude of the peak value in the potential energy profiles of the nTiO2colloids is4.2times that of the nCeO2colloids, indicating that nTiO2colloids displayed more stable than the nCeO2colloids. The nTiO2colloids inhibited the cell membrane permeability of Scenedesmus obliquus, conversely, the nCeO2colloids induced an enhancement of the cell membrane permeability of Scenedesmus obliquus. The two nanoparticle colloids induced an increase of the cell membrane potential of Scenedesmus ohliquus, and the nTiO2colloids showed stronger effects than the nCcO2colloids. These findings suggest that disturbing the functions of the cell membrane is a possible mechanism of toxicity of MNPs to algae. Compared with the nCeO2colloids, the nTiO2colloids with high stability displayed high toxicity to the cell membrane system in Scenedesmus obliquus, suggesting that the colloid stability of MNPs is an important factor governing their ecotoxcitity.
(3) The relative contribution of ionic silver (Ag+) or particles in nano-silver (nAg) colloids to the toxicity to three aquatic organisms of different trophic levels, i.e. an alga species (Raphidocelis subcapitata), a cladoceran species (Chydorus sphaericus) and a freshwater fish larva (Dainio rerio), was quantitatively evaluated by a response addition model. A bare and a polyvinylpyrrolidone (PVP)-coated nAg, as well as a monodispersed nAg with a dispersant (DIS-nAg) were examined. The toxicity of the nAg in the form of colloids decreases in the order DIS-nAg> PVP-nAg> Bare-nAg for all the three trophic aquatic organisms (in terms of median effect concentration). The DIS-nAg had the highest free Ag+concentration. and the Bare-nAg the lowest concentration of free Ag+, implying that the free Ag+cannot be neglected in explaining the toxicity of the nAg colloids. Furthermore, it was found that the contribution of free Ag+to the toxicity of nAg colloids for Raphidocelis subcapitata is the highest, but for Danio rerio the lowest, implying that the organisms tested have different accumulation ability for Ag+or nAg particles.
(4) The effects of DOM on the ecotoxicity of nAg colloids were investigated. A commercial DOM model compound (HS) and PVP-coated nAg with primary size of20nm were selected as test materials. Raphidocelis subcapitata, Chydorus sphaericus, and Danio rerio, representing different trophic organisms, were exposed to the nAg colloids in the presence and absence of HS. The results show that the presence of HS alleviates the aquatic toxicity of the nAg colloids to all the organisms in a concentration dependent manner. A particle size distribution shifts to lower values of the nAg colloidal particles due to the presence of HS, implying that the decrease in toxicity of the nAg colloids is not related to the variation of agglomerated size. Surface charge of nAg agglomerates only displayed more negative in the presence of HS with high concentration, suggesting that the surface charge is not a decisive factor for the decrease in the toxicity of the nAg colloids. The presence of HS inhibited Ag'release from the nAg particles, and the inhibition degree depended on the HS concentrations. In addition, a concentration-response relationship was observed clearly for the toxicity effects expressed as free Ag+in the colloids. This finding indicates that the HS controlling Ag'release is a crucial mechanism, which contributes to the detoxification of nAg released to the environment.
(1)利用分子力学和量子化学计算,模拟了C60与不同来源的DOM模型分子间的相互作用。相互作用能的计算结果表明,DOM模型分子可与C60间形成稳定的复合物。与来源于淡水或褐煤的DOM模型分子相比较,土壤中的DOM模型分子与C60间形成的复合物的稳定性更高。电荷转移分析的结果表明,DOM在C60-DOM的复合物中是电子受体。此外,DOM的存在显著地提高了C60的表观水溶解度(SC60/DOM),且SC60/DOM随C60-DOM复合物的前线分子轨道能级差的增加而降低。
(2)采用荧光探针法研究了nTiO2和nCeO2胶体对斜生栅藻(Scenedesmus obliquus)细胞膜通透性和膜电位的影响。利用DLVO理论对这两种MNPs的胶体稳定性进行了评价。在总势能曲线上nTiO2胶体的势能峰值是nCeO2的4.2倍,表明nTiO2胶体的稳定性高于nCeO2胶体。nTiO2胶体降低了绿藻细胞膜通透性,nCeO2胶体却增加了绿藻细胞膜通透性。这两种MNPs胶体均引起细胞膜电位的升高,且nTiO2胶体诱导细胞膜电位升高程度明显高于nCeO2胶体,表明干扰绿藻细胞膜的正常生理功能是MNPs对绿藻的毒性作用机制之一。与nCeO2胶体相比,具有更高悬浮稳定性的nTiO2胶体表现出更强的绿藻细胞膜毒性效应,说明MNPs的胶体稳定性是决定其毒性差异的重要因素。
(3)考察了nAg胶体对三种不同营养级水生生物的毒理效应,包括羊角月牙藻(Raphidocelis subcapitata)、圆形盘肠溞(Chydorus sphaericus)及斑马鱼(Danio rerio)胚胎。通过反应加和模型定量评价了游离态银离子(Ag+)对nAg胶体的毒性贡献。研究了三种商业nAg颗粒物：无包覆剂(Bare)nAg、聚乙烯吡咯烷酮包覆(PVP)nAg、悬浮剂分散(DIS)nAg。发现这三种nAg胶体对受试生物的毒性(以半数效应浓度为基础)大小顺序均为：DIS-nAg> PVP-nAg> Bare-nAg。DIS-nAg含有最高浓度的游离态Ag+,而Bare-nAg中的游离态Ag+的浓度最低,表明主解释nAg的毒性机制时不能忽略游离态Ag+的作用。此外,游离态Ag+对nAg胶体的Raphidocelis subcapitata毒性贡献最大,而对nAg胶体的Danio rerio毒性贡献最小,说国游离态Ag+对nAg颗粒对不同水生生物的毒性贡献不同,取决于水生生物的营养级水平。
(4)研究了DOM对nAg胶体水生毒性的影响。以一种腐殖酸模型分子(HS)来模拟DOM,考察了HS对PVP包覆的nAg(一次粒径为20nm)对羊角月牙藻、圆形盘肠溞及斑马鱼胚胎的毒性影响。发现随HS浓度的升高,nAg胶体对三种水生生物的毒性降低。随着HS浓度的增加,nAg胶体颗粒物的尺度更小,意味颗粒物的尺度变化不是导致HS存在条件下nAg毒性降低的原因。HS浓度增高时,导致nAg表面电荷更负,说明表面电荷大小也不是导致有HS存在时nAg毒性降低的决定因素；HS浓度较高时,Ag+释放受到了抑制,以nAg胶体中游离态Ag+浓度来表达胶体的毒性反应时,存在明显的浓度-反应关系,说明HS通过抑制Ag+的释放来降低nAg胶体的水生毒性。
With the increasing use of various nanostructured materials, manufacuted nanoparticles (MNPs) are inevitablily discharged into the environment. Thereby MNPs could become potential pollutants, and pose a potential health risk to human and ecological species. Once released into the environment, MNPs can undergo many transport and transformation processes affecting their ecotoxicity. How to assess and predict aquantic behavior (particular for the interaction of MNPs with dissolved organic matter (DOM) and their effects on agglomeration of MNPs colloids) and toxicological effects to aquatic organisms is the frontier research in the field of nanoecotoxicology. In this thesis, interactions of fullerene (C6o) with dissolved organic matters (DOM) were investigated by the means of molecular modelling, and the mechanism by which DOM influence dissolution and agglomeration behavior of C6o was elucidated. In addition, toxicological effects of nano-titanium dioxide (nTiO2), nano-cerium dioxide (nCeO2) and nano-silver (nAg) colloids on aquatic organisms were observed and mechanisms of action for their aquatic toxicity were illustrated. Main contents and results are as follows:
(1) The interaction of fullerene C60with DOM analogues is computationally simulated by molecular mechanics and density functional theory. The calculated interaction energies displayed that the DOM could stablize C60, and the stability between C60and the soil DOM analogues is greater than that between C60and the DOM analogues from fresh water or coal. The computed electrostatic potential indicates that DOM analogues are electron acceptors in the C60-DOM complexes. The presence of DOM increases the apparent water solubility of C60-It is also observed that the C60apparent water solubility decreases with the increase of the energy gaps of frontier molecular orbitals (ELUMO-EHOMO) for each C60-DOM complex.
(2) The effects of nTiO2and nCeO2colloids on cell membrane permeability and cell membrane potential of a green alga (Scenedesmus obliquus) were investigated by the fluorescent probe technique. The stability of nanoparticle colloids was evaluated by the DLVO theory. The results show that the magnitude of the peak value in the potential energy profiles of the nTiO2colloids is4.2times that of the nCeO2colloids, indicating that nTiO2colloids displayed more stable than the nCeO2colloids. The nTiO2colloids inhibited the cell membrane permeability of Scenedesmus obliquus, conversely, the nCeO2colloids induced an enhancement of the cell membrane permeability of Scenedesmus obliquus. The two nanoparticle colloids induced an increase of the cell membrane potential of Scenedesmus ohliquus, and the nTiO2colloids showed stronger effects than the nCcO2colloids. These findings suggest that disturbing the functions of the cell membrane is a possible mechanism of toxicity of MNPs to algae. Compared with the nCeO2colloids, the nTiO2colloids with high stability displayed high toxicity to the cell membrane system in Scenedesmus obliquus, suggesting that the colloid stability of MNPs is an important factor governing their ecotoxcitity.
(3) The relative contribution of ionic silver (Ag+) or particles in nano-silver (nAg) colloids to the toxicity to three aquatic organisms of different trophic levels, i.e. an alga species (Raphidocelis subcapitata), a cladoceran species (Chydorus sphaericus) and a freshwater fish larva (Dainio rerio), was quantitatively evaluated by a response addition model. A bare and a polyvinylpyrrolidone (PVP)-coated nAg, as well as a monodispersed nAg with a dispersant (DIS-nAg) were examined. The toxicity of the nAg in the form of colloids decreases in the order DIS-nAg> PVP-nAg> Bare-nAg for all the three trophic aquatic organisms (in terms of median effect concentration). The DIS-nAg had the highest free Ag+concentration. and the Bare-nAg the lowest concentration of free Ag+, implying that the free Ag+cannot be neglected in explaining the toxicity of the nAg colloids. Furthermore, it was found that the contribution of free Ag+to the toxicity of nAg colloids for Raphidocelis subcapitata is the highest, but for Danio rerio the lowest, implying that the organisms tested have different accumulation ability for Ag+or nAg particles.
(4) The effects of DOM on the ecotoxicity of nAg colloids were investigated. A commercial DOM model compound (HS) and PVP-coated nAg with primary size of20nm were selected as test materials. Raphidocelis subcapitata, Chydorus sphaericus, and Danio rerio, representing different trophic organisms, were exposed to the nAg colloids in the presence and absence of HS. The results show that the presence of HS alleviates the aquatic toxicity of the nAg colloids to all the organisms in a concentration dependent manner. A particle size distribution shifts to lower values of the nAg colloidal particles due to the presence of HS, implying that the decrease in toxicity of the nAg colloids is not related to the variation of agglomerated size. Surface charge of nAg agglomerates only displayed more negative in the presence of HS with high concentration, suggesting that the surface charge is not a decisive factor for the decrease in the toxicity of the nAg colloids. The presence of HS inhibited Ag'release from the nAg particles, and the inhibition degree depended on the HS concentrations. In addition, a concentration-response relationship was observed clearly for the toxicity effects expressed as free Ag+in the colloids. This finding indicates that the HS controlling Ag'release is a crucial mechanism, which contributes to the detoxification of nAg released to the environment.
引文
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