近海环境中天然有机质的分离与表征
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摘要
近海环境中的天然有机质(包括沉积物有机质、颗粒态有机质和溶解有机质)是全球碳循环中的重要组成部分,对全球生物地球化学循环以及区域生态系统对气候变暖的响应有着重要的作用。在海洋环境中,它们不但可以为海洋中异养型微生物提供营养和能量,促进它们在海洋中的循环,还能对海水的pH值起到调节和缓冲作用,影响海水对光的吸收,从而影响海水的颜色和表层温度。海洋天然有机质还能与海洋环境中的金属离子、有机污染物形成配合物,从而影响它们在海洋中的毒性和生物利用性。
由于海洋天然有机质本身结构组成的复杂性,以及对其有效分离的困难性,人们对它们的性质以及地球化学行为的认知至今仍然有限。本文着重近海环境中沉积物有机质和水体溶解态有机质的分离和表征的研究,分别采用国际腐殖质协会推荐的方法和一种新颖的反渗透耦合电渗析的方法从近岸沉积物和海水中分离出了沉积物腐殖质、溶解态腐殖质以及溶解有机质。采用多种化学和光谱表征手段,包括元素分析、稳定同位素分析、傅里叶变换红外光谱分析、热重分析、固态13C核磁共振分析、傅里叶变换离子回旋共振质谱分析、电位滴定分析等对所分离的近海天然有机质进行了分析和比较。以期能够对所研究天然有机质的来源、组成、反应性和归宿等科学问题进行阐述,并对采用不同方法所分离的海洋溶解有机质的性质做一比较。
通过对长江口沉积物腐殖质以及海南东部红树林沉积物腐殖质的化学和光谱性质的研究发现：近岸以及河口沉积物腐殖质是沉积物有机质的主要组成部分,其中胡敏素在沉积物有机质中所占的比例最大,然后依次是腐殖酸和富里酸。比较富里酸和腐殖酸的性质发现,腐殖酸中含有更多的含氮化合物及高度支链化的长链脂肪族化合物,而富里酸中则含有更多的含氧官能团,比如羧基和醇类物质等。随着离岸距离的增加或海源物质贡献的增大,腐殖质中H/C原子比、δ13C值以及脂肪族化合物、蛋白质、碳水化合物组分的含量逐渐增大,而C/N原子比、芳香族化合物和酸性官能团的含量则逐渐减少。通过对长江口不同层次腐殖质的研究发现,腐殖质中酸性官能团和芳香族化合物的含量是随着沉积物深度的增加而逐渐增加的,而其中碳水化合物的含量却是随深度的增加而逐渐减少的。选择性的保留木质素等顽固组分而消耗易被微生物所利用的反应性组分(如碳水化合物和蛋白质等)可能是长江口沉积物有机质早期成岩反应的主要过程。本文所研究沉积物腐殖质中质子结合位点的平均浓度大约为22meq gC-1,这一数值也许能为所研究地区沉积物中金属离子的分布形态提供一定的科学依据。
通过对海南东部红树林间隙水及其下游近岸海水中溶解态腐殖质性质的研究发现：相比较于陆源物质衍生的红树林间隙水溶解腐殖质,近岸海水溶解腐殖质中含有更多的13C同位素,脂肪族化合物,碳水化合物,但芳香族化合物和富含羧基的化合物含量较少。近岸海水溶解腐殖质可能为光降解的红树林溶解腐殖质和海洋溶解腐殖质的混合物。结合红树林沉积物腐殖质的δ13C和C/N比数据可以发现,红树林地区的沉积物有机质和溶解有机质大部分来源于当地红树林碎屑的贡献,并能对周围的环境产生较大的影响。本文通过高分辨率质谱分析发现,所研究溶解腐殖质中各个级分之间的分子组成存在较大的差异,因而它们可能是处于不同腐殖化阶段的产物。富里酸可能较腐殖酸要老一些,而亲水性的酸性组分XAD-4部分可能为最老的部分。木质素降解反应和Maillard缩合反应的共同作用可能是该地区溶解腐殖质生成和转化的主要机理。
对采用反渗透耦合电渗析(]RO/ED)的方法从Barataria湾和大西洋近岸海水中所分离溶解有机质的性质进行研究发现：采用RO/ED法所分离的海洋溶解有机质中羧基含量约为6.0～6.9meq gC-1,大约只为土壤及河流天然有机质及富里酸中羧基含量的60%左右,且所研究海洋溶解有机质中没有明显的酚羟基存在。表层海水溶解有机质中碳水化合物,蛋白质,以及类脂物质的含量较高,而底层海水溶解有机质中芳香族化合物,以及缩合的碳氢化合物等物质的含量较高。说明多糖、蛋白质等物质是海洋溶解有机质中的活性组分,可能是海洋中异养型微生物获得能量的主要来源。通过与文献中所报道的由XAD树脂法和超滤法所获得的海洋溶解有机质性质的相互比较可以发现,RO/ED法即可获得超滤法容易分离的碳水化合物组分,又可获得固相萃取法容易获得的富含羧基的组分以及脂肪族化合物,从而说明RO/ED法可能能够从海水中分离出具有代表性的海洋溶解有机质。
Natural organic matter (NOM) in the coastal environments, which include the sedimentary organic matters (SOM), particulate organic matters (POM), and dissolved organic matters (DOM), is an important component of global carbon cycle and plays important roles in the response to climate change. The bioavailable fraction of NOM can be consumed by heterotrophic microbes, and is an important component in the marine microbial cycle. The colored fraction of DOM is the principal chromophore in marine waters, influencing the color and temperature of surface waters. Marine NOM also plays key roles in metal and organic pollutants chelation, influencing metal and organic toxicity arid bioavailability.
Due to the extremely complexity of marine NOM and the difficulty to acquire representative marine NOM samples, our knowledges for their properties and geochemical behaviors are still limited. This dissertation mainly focuses on the isolation and characterization of SOM and DOM in coastal environments. The sedimentary humic substances (SHS), dissolved humic substances (DHS) and DOM were isolated by using the methods established by the International Humic Substances Society and a novel coupled reverse osmosis-electrodialysis (RO/ED) method, respectively. To better understand the origin, composition, reactivity, and fate of the coastal NOM, elemental analysis, stable isotope analysis, Fourier transformed infrared spectroscopy (FTIR), thermogravimetric analysis (TGA),13C nuclear magnetic resonance spectroscopy (13C NMR), Fourier transform ion cyclotron resonance mass spectrometry (FTICR-MS) and potentiometric titrations were used to analyze those isolated samples. The different properties of marine DOM which were isolated by different methods were also compared and analyzed.
The results of the SHS which were isolated from the estuarine and coastal sediments of the East China Sea as well as the mangrove sediments in the eastern coast of Hainan Island indicated that SHS represented a large fraction of SOM, with humin were the dominant fraction of SHS followed by humic acids and fulvic acids. Humic acids were found contain more nitrogen-containing compounds and highly branched long-chain aliphatic moieties, while fulvic acids contain more oxygen-containing functional groups, such as carboxyl and alcoholic groups. With the increasing distance from the shore and/or the increasing contribution from the marine source, the H/C ratios,δ13C values, as well as the content of aliphatic compounds, proteins, and carbohydrates increased, while the C/N ratios as well as the content of aromatic compounds and acidic functional groups decreased. As for the depth profile of SHS samples in the Changjiang estuary, it is found that the charge densities and aromatic compounds increased with increasing depth; however, the content of carbohydrates decreased with increasing depth. Therefore, it probably can be inferred that loss of labile carbohydrates and proteins and selective preservation of refractory lignin components may dominate the early diagenetic reactions of SHS in the Changjiang estuary. The average concentration of binding sites for the studied SHS is about22meq gC-1, and this value may provide insights for the species of metal cations in the sediments.
The properties of DHS which were isolated from one mangrove pore-water sample and one near shore seawater sample downstream the mangrove-fringed estuary in the eastern coastal of Hainan Island indicated that the near shore seawater DHS enrich in13C and contain more aliphatic compounds, carbohydrates, but less aromatic structures and carboxyl groups than that of mangrove pore-water DHS. The significant mixing of photo-degraded mangrove DHS and the marine DHS were considered to the mechanism results in the difference for the two DHS samples. Combined with the δ13C values and C/N ratios of mangrove SHS in our previous study, it can be found both SOM and DOM in mangrove area are mainly from autochthonous mangrove detritus and can contribute to ambient environment in a large scale. The results of FTICR-MS indicated that there are big differences among the different fractions of DHS in the studied water samples, which may attribute to the different humification level of the different fractions. Fulvic acids may older than humic acids, while the hydrophilic XAD-4fraction may be the oldest fraction among DHS. The combination of the lignin degradation reaction and the Maillard condensation reactions are probably the formation and transformation mechanism of the studied DHS samples.
The RO/ED method was used to isolate marine DOM from Barataria Bay and the coastal of Atlantic Ocean. The acid-base properties of the isolated marine DOM indicated that the concentration of carboxyl groups in marine DOM samples is about6.0～6.9meq gC-1, which is only60%of the carboxyl contents for soil and river NOM and fulvic acids. Phenolic hydroxyl groups could not be detected in the studied samples. Marine DOM in surface waters contains more carbohydrates, proteins, and lipids; however, marine DOM in deeper waters contains more aromatic compounds and condensed hydrocarbons. This phenomenon indicated that carbohydrates and proteins are reactive components, perhaps supporting much of the heterotrophic activity in the surface ocean. By comparing the properties of marine DOM isolated by RO/ED with that isolated by ultrafiltration and XAD resins which were reported in literatures, it can be found that the RO/ED method recovers both the carbohydrate-rich fraction of DOM that is isolated preferentially by UF and the fraction of DOM that is enriched in carboxyl groups and aliphatic moieties that is isolated preferentially by SPE methods. So the RO/ED method may recovery representative marine DOM from sea water.
由于海洋天然有机质本身结构组成的复杂性,以及对其有效分离的困难性,人们对它们的性质以及地球化学行为的认知至今仍然有限。本文着重近海环境中沉积物有机质和水体溶解态有机质的分离和表征的研究,分别采用国际腐殖质协会推荐的方法和一种新颖的反渗透耦合电渗析的方法从近岸沉积物和海水中分离出了沉积物腐殖质、溶解态腐殖质以及溶解有机质。采用多种化学和光谱表征手段,包括元素分析、稳定同位素分析、傅里叶变换红外光谱分析、热重分析、固态13C核磁共振分析、傅里叶变换离子回旋共振质谱分析、电位滴定分析等对所分离的近海天然有机质进行了分析和比较。以期能够对所研究天然有机质的来源、组成、反应性和归宿等科学问题进行阐述,并对采用不同方法所分离的海洋溶解有机质的性质做一比较。
通过对长江口沉积物腐殖质以及海南东部红树林沉积物腐殖质的化学和光谱性质的研究发现：近岸以及河口沉积物腐殖质是沉积物有机质的主要组成部分,其中胡敏素在沉积物有机质中所占的比例最大,然后依次是腐殖酸和富里酸。比较富里酸和腐殖酸的性质发现,腐殖酸中含有更多的含氮化合物及高度支链化的长链脂肪族化合物,而富里酸中则含有更多的含氧官能团,比如羧基和醇类物质等。随着离岸距离的增加或海源物质贡献的增大,腐殖质中H/C原子比、δ13C值以及脂肪族化合物、蛋白质、碳水化合物组分的含量逐渐增大,而C/N原子比、芳香族化合物和酸性官能团的含量则逐渐减少。通过对长江口不同层次腐殖质的研究发现,腐殖质中酸性官能团和芳香族化合物的含量是随着沉积物深度的增加而逐渐增加的,而其中碳水化合物的含量却是随深度的增加而逐渐减少的。选择性的保留木质素等顽固组分而消耗易被微生物所利用的反应性组分(如碳水化合物和蛋白质等)可能是长江口沉积物有机质早期成岩反应的主要过程。本文所研究沉积物腐殖质中质子结合位点的平均浓度大约为22meq gC-1,这一数值也许能为所研究地区沉积物中金属离子的分布形态提供一定的科学依据。
通过对海南东部红树林间隙水及其下游近岸海水中溶解态腐殖质性质的研究发现：相比较于陆源物质衍生的红树林间隙水溶解腐殖质,近岸海水溶解腐殖质中含有更多的13C同位素,脂肪族化合物,碳水化合物,但芳香族化合物和富含羧基的化合物含量较少。近岸海水溶解腐殖质可能为光降解的红树林溶解腐殖质和海洋溶解腐殖质的混合物。结合红树林沉积物腐殖质的δ13C和C/N比数据可以发现,红树林地区的沉积物有机质和溶解有机质大部分来源于当地红树林碎屑的贡献,并能对周围的环境产生较大的影响。本文通过高分辨率质谱分析发现,所研究溶解腐殖质中各个级分之间的分子组成存在较大的差异,因而它们可能是处于不同腐殖化阶段的产物。富里酸可能较腐殖酸要老一些,而亲水性的酸性组分XAD-4部分可能为最老的部分。木质素降解反应和Maillard缩合反应的共同作用可能是该地区溶解腐殖质生成和转化的主要机理。
对采用反渗透耦合电渗析(]RO/ED)的方法从Barataria湾和大西洋近岸海水中所分离溶解有机质的性质进行研究发现：采用RO/ED法所分离的海洋溶解有机质中羧基含量约为6.0～6.9meq gC-1,大约只为土壤及河流天然有机质及富里酸中羧基含量的60%左右,且所研究海洋溶解有机质中没有明显的酚羟基存在。表层海水溶解有机质中碳水化合物,蛋白质,以及类脂物质的含量较高,而底层海水溶解有机质中芳香族化合物,以及缩合的碳氢化合物等物质的含量较高。说明多糖、蛋白质等物质是海洋溶解有机质中的活性组分,可能是海洋中异养型微生物获得能量的主要来源。通过与文献中所报道的由XAD树脂法和超滤法所获得的海洋溶解有机质性质的相互比较可以发现,RO/ED法即可获得超滤法容易分离的碳水化合物组分,又可获得固相萃取法容易获得的富含羧基的组分以及脂肪族化合物,从而说明RO/ED法可能能够从海水中分离出具有代表性的海洋溶解有机质。
Natural organic matter (NOM) in the coastal environments, which include the sedimentary organic matters (SOM), particulate organic matters (POM), and dissolved organic matters (DOM), is an important component of global carbon cycle and plays important roles in the response to climate change. The bioavailable fraction of NOM can be consumed by heterotrophic microbes, and is an important component in the marine microbial cycle. The colored fraction of DOM is the principal chromophore in marine waters, influencing the color and temperature of surface waters. Marine NOM also plays key roles in metal and organic pollutants chelation, influencing metal and organic toxicity arid bioavailability.
Due to the extremely complexity of marine NOM and the difficulty to acquire representative marine NOM samples, our knowledges for their properties and geochemical behaviors are still limited. This dissertation mainly focuses on the isolation and characterization of SOM and DOM in coastal environments. The sedimentary humic substances (SHS), dissolved humic substances (DHS) and DOM were isolated by using the methods established by the International Humic Substances Society and a novel coupled reverse osmosis-electrodialysis (RO/ED) method, respectively. To better understand the origin, composition, reactivity, and fate of the coastal NOM, elemental analysis, stable isotope analysis, Fourier transformed infrared spectroscopy (FTIR), thermogravimetric analysis (TGA),13C nuclear magnetic resonance spectroscopy (13C NMR), Fourier transform ion cyclotron resonance mass spectrometry (FTICR-MS) and potentiometric titrations were used to analyze those isolated samples. The different properties of marine DOM which were isolated by different methods were also compared and analyzed.
The results of the SHS which were isolated from the estuarine and coastal sediments of the East China Sea as well as the mangrove sediments in the eastern coast of Hainan Island indicated that SHS represented a large fraction of SOM, with humin were the dominant fraction of SHS followed by humic acids and fulvic acids. Humic acids were found contain more nitrogen-containing compounds and highly branched long-chain aliphatic moieties, while fulvic acids contain more oxygen-containing functional groups, such as carboxyl and alcoholic groups. With the increasing distance from the shore and/or the increasing contribution from the marine source, the H/C ratios,δ13C values, as well as the content of aliphatic compounds, proteins, and carbohydrates increased, while the C/N ratios as well as the content of aromatic compounds and acidic functional groups decreased. As for the depth profile of SHS samples in the Changjiang estuary, it is found that the charge densities and aromatic compounds increased with increasing depth; however, the content of carbohydrates decreased with increasing depth. Therefore, it probably can be inferred that loss of labile carbohydrates and proteins and selective preservation of refractory lignin components may dominate the early diagenetic reactions of SHS in the Changjiang estuary. The average concentration of binding sites for the studied SHS is about22meq gC-1, and this value may provide insights for the species of metal cations in the sediments.
The properties of DHS which were isolated from one mangrove pore-water sample and one near shore seawater sample downstream the mangrove-fringed estuary in the eastern coastal of Hainan Island indicated that the near shore seawater DHS enrich in13C and contain more aliphatic compounds, carbohydrates, but less aromatic structures and carboxyl groups than that of mangrove pore-water DHS. The significant mixing of photo-degraded mangrove DHS and the marine DHS were considered to the mechanism results in the difference for the two DHS samples. Combined with the δ13C values and C/N ratios of mangrove SHS in our previous study, it can be found both SOM and DOM in mangrove area are mainly from autochthonous mangrove detritus and can contribute to ambient environment in a large scale. The results of FTICR-MS indicated that there are big differences among the different fractions of DHS in the studied water samples, which may attribute to the different humification level of the different fractions. Fulvic acids may older than humic acids, while the hydrophilic XAD-4fraction may be the oldest fraction among DHS. The combination of the lignin degradation reaction and the Maillard condensation reactions are probably the formation and transformation mechanism of the studied DHS samples.
The RO/ED method was used to isolate marine DOM from Barataria Bay and the coastal of Atlantic Ocean. The acid-base properties of the isolated marine DOM indicated that the concentration of carboxyl groups in marine DOM samples is about6.0～6.9meq gC-1, which is only60%of the carboxyl contents for soil and river NOM and fulvic acids. Phenolic hydroxyl groups could not be detected in the studied samples. Marine DOM in surface waters contains more carbohydrates, proteins, and lipids; however, marine DOM in deeper waters contains more aromatic compounds and condensed hydrocarbons. This phenomenon indicated that carbohydrates and proteins are reactive components, perhaps supporting much of the heterotrophic activity in the surface ocean. By comparing the properties of marine DOM isolated by RO/ED with that isolated by ultrafiltration and XAD resins which were reported in literatures, it can be found that the RO/ED method recovers both the carbohydrate-rich fraction of DOM that is isolated preferentially by UF and the fraction of DOM that is enriched in carboxyl groups and aliphatic moieties that is isolated preferentially by SPE methods. So the RO/ED method may recovery representative marine DOM from sea water.
引文
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