Extracellular polymeric substances

细胞外聚合物质
  • 文章类型: Journal Article
    胞外聚合物质(EPS)已显示出减少多价金属污染的显著益处。使用从中国煤化工场所分离出的木氧无色杆菌BP1,这项研究阐明了EPS产生对Cr(VI)还原的贡献,并揭示了其生物去除机理。BP1在最适pH为8时生长,Cr(VI)的最低抑制浓度为300mg/L。废培养基完全去除Cr(VI),静息细胞只能去除10.47%,失活细胞几乎不能去除Cr(VI)。S-EPS和B-EPS降低Cr(VI)98.59%和11.64%,分别。SEM-EDS分析表明,在Cr胁迫下,BP1细胞被刺激产生EPS。XPS结果表明,通过细胞内生物富集或生物吸附富集了29.63%的Cr(VI),通过细胞外基质酶还原了70.37%的Cr(VI),产生了Cr(OH)3和有机Cr(III)复合物。根据FTIR,带有-OH的EPS,COO-,和酰胺基团为Cr(VI)的还原吸附提供了结合位点和电子。基因组研究表明,BP1主要产生胞外多糖,代谢硫和氮,并减少DNA修复蛋白酶导致的活性氧损伤。
    Extracellular polymeric substances (EPS) have demonstrated significant benefits for reducing multivalent metal contamination. Using Achromobacter xylosoxidans BP1 isolated from a coal chemical site in China, this study elucidated the contribution of EPS production to Cr (VI) reduction and revealed its biological removal mechanism. BP1 grew at an optimum pH of 8 and the lowest inhibitory concentration of Cr(VI) was 300 mg/L. The spent medium completely removed Cr(VI), whereas resting cells were only able to remove 10.47 % and inactivated cells were nearly incapable of Cr(VI) removal. S-EPS and B-EPS reduced Cr(VI) by 98.59 % and 11.64 %, respectively. SEM-EDS analysis showed that the BP1 cells were stimulated to produce EPS under Cr stress. The XPS results showed that 29.63 % of Cr(VI) was enriched by intracellular bioaccumulation or biosorption and 70.37 % of Cr(VI) was reduced by extracellular enzymes to produce Cr(OH)3 and organic Cr(III) complexes. According to FTIR, EPS with -OH, COO-, and amide groups supplied binding sites and electrons for the reductive adsorption of Cr(VI). Genomic studies showed that BP1 primarily produces extracellular polysaccharides, metabolises sulphur and nitrogen, and reduces reactive oxygen species damage as a result of DNA repair proteases.
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  • 文章类型: Journal Article
    在污泥脱水过程中,由于胞外聚合物(EPS)和结合水之间的强烈相互作用,因此出现了严峻的挑战。这项研究介绍了一部小说,采用铁(Fe2+)/过氧单硫酸盐(PMS)和聚丙烯酰胺(PAM)协同调理的方法显著提高污泥脱水效率。Fe2+/PMS-PAM调理方法的应用导致比过滤阻力(SFR)显著降低82.75%,毛细管抽吸时间(CST)显著降低80.44%,标志着脱水性能的显著提高。综合分析表明,Fe2+/PMS-PAM过程中Fe2+/PMS预氧化能有效降解EPS,促进结合水的释放。随后,PAM增强了高级氧化过程(AOPs)产生的细污泥颗粒的絮凝。此外,基于扩展的Derjaguin-Landau-Verwey-Overbeek(XDLVO)理论的分析证明了相互作用能的变化,强调污泥内能量屏障的破坏和表面特征从亲水性(3.79mJm-2)到疏水性(-61.86mJm-2)的转变。这种转变促进了污泥颗粒的自发聚集。Flory-Huggins理论的创新使用从化学势的角度提供了对污泥过滤机理的见解,将这些更改链接到SFR。Fe2+/PMS-PAM调节的引入破坏了EPS形成的凝胶层的均匀性,显着降低渗透物和凝胶层中的水之间的化学电位差,导致较低的SFR和增强的脱水性能。这种热力学方法大大增强了我们对污泥脱水和调节的理解。这些发现代表了范式的转变,为污泥处理提供创新策略,扩大我们对脱水和调节技术的理解。
    In the sludge dewatering process, a formidable challenge arises due to the robust interactions between extracellular polymeric substances (EPS) and bound water. This study introduces a novel, synergistic conditioning method that combines iron (Fe2+)/peroxymonosulfate (PMS) and polyacrylamide (PAM) to significantly enhance sludge dewatering efficiency. The application of the Fe2+/PMS-PAM conditioning method led to a substantial reduction in specific filtration resistance (SFR) by 82.75% and capillary suction time (CST) by 80.44%, marking a considerable improvement in dewatering performance. Comprehensive analyses revealed that pre-oxidation with Fe2+/PMS in the Fe2+/PMS-PAM process effectively degraded EPS, facilitating the release of bound water. Subsequently, PAM enhanced the flocculation of fine sludge particles resulting from the advanced oxidation processes (AOPs). Furthermore, analysis based on the Extended Derjaguin-Landau-Verwey-Overbeek (XDLVO) theory demonstrated shifts in interaction energies, highlighting the breakdown of energy barriers within the sludge and a transition in surface characteristics from hydrophilic (3.79 mJ m-2) to hydrophobic (-61.86 mJ m-2). This shift promoted the spontaneous aggregation of sludge particles. The innovative use of the Flory-Huggins theory provided insights into the sludge filtration mechanism from a chemical potential perspective, linking these changes to SFR. The introduction of Fe2+/PMS-PAM conditioning disrupted the uniformity of the EPS-formed gel layer, significantly reducing the chemical potential difference between the permeate and the water in the gel layer, leading to a lower SFR and enhanced dewatering performance. This thermodynamic approach significantly enhances our understanding of sludge dewatering and conditioning. These findings represent a paradigm shift, offering innovative strategies for sludge treatment and expanding our comprehension of dewatering and conditioning techniques.
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  • 文章类型: Journal Article
    废弃的工业场地和管理不当的废物处置区的铬污染构成了重大的环境威胁。微生物诱导碳酸盐沉淀(MICP)显示出很有希望,修复Cr(VI)和二价重金属的生态友好解决方案。在这项研究中,MICP是通过尿素分解细菌肌酸杆菌进行铬固定的,该细菌能够通过细胞外聚合物(EPS)的产生将Cr(VI)还原为毒性较低的Cr(III)。通过细胞部分分析证实了EPS驱动的减少的功效。MICP在水溶液中与100ppm的Cr(VI)共沉淀82.21%的铬与CaCO3,共沉淀与Cr(VI)的还原呈正相关。该生物用于修复掺加铬的砂,发现与对照相比,MICP处理将铬的可交换分数降低至0.54±0.11%,并将碳酸盐结合分数提高至26.1±1.15%。XRD和SEM分析显示还原过程中产生的Cr(III),影响了沉淀过程中球文石的多晶型选择。评价MICP修复从Ranipet收集的Cr污染土壤样品,泰米尔纳德邦也显示出有效的铬固定化。因此,溶肌酸杆菌被证明是通过MICP工艺包裹铬污染土壤的可行选择,有效缓解了Cr(VI)向地下水及邻近水体的渗入。
    Chromium contamination from abandoned industrial sites and inadequately managed waste disposal areas poses substantial environmental threat. Microbially induced carbonate precipitation (MICP) has shown promising, eco-friendly solution to remediate Cr(VI) and divalent heavy metals. In this study, MICP was carried out for chromium immobilization by an ureolytic bacterium Arthrobacter creatinolyticus which is capable of reducing Cr(VI) to less toxic Cr(III) via extracellular polymeric substances (EPS) production. The efficacy of EPS driven reduction was confirmed by cellular fraction analysis. MICP carried out in aqueous solution with 100 ppm of Cr(VI) co-precipitated 82.21% of chromium with CaCO3 and the co-precipitation is positively correlated with reduction of Cr(VI). The organism was utilized to remediate chromium spiked sand and found that MICP treatment decreased the exchangeable fraction of chromium to 0.54 ± 0.11% and increased the carbonate bound fraction to 26.1 ± 1.15% compared to control. XRD and SEM analysis revealed that Cr(III) produced during reduction, influenced the polymorph selection of vaterite during precipitation. Evaluation of MICP to remediate Cr polluted soil sample collected from Ranipet, Tamil Nadu also showed effective immobilization of chromium. Thus, A. creatinolyticus proves to be viable option for encapsulating chromium contaminated soil via MICP process, and effectively mitigating the infiltration of Cr(VI) into groundwater and adjacent water bodies.
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  • 文章类型: Journal Article
    生物体分泌的细胞外聚合物(EPS)倾向于包裹微塑料(MPs),形成EPS日冕,影响海洋生态系统中MP的命运。然而,EPS电晕对MP对海洋生物的生物毒性的影响仍然知之甚少。在这里,EPS电晕对不同尺寸(0.1和1µm)的聚苯乙烯(PS)MPs对Skeletonemacostatum的毒性的影响(S.costatum)进行了调查。中等分子量(〜55kDa)蛋白质在PSMPs上的优先吸附主要有助于EPS电晕的形成,将小型PSMPs(0.1µm)的表面电荷负性降低72.4%。多糖和蛋白质中的氮(N)和氧(O)部分被确定为EPS-PSMPs相互作用中的优先吸附位点。密度泛函理论(DFT)计算证实了核磁共振波谱(NMR)的结果,表明EPS与PSMPs之间的结合模式主要为氢键。此外,当暴露于小尺寸PSMPs(0.1µm,25-50mg/L)。这些发现为EPS电晕如何影响海洋生态系统中与微米和纳米级塑料相关的环境命运和生态风险提供了新的见解。
    Extracellular polymeric substances (EPS) secreted by organisms tend to encapsulate microplastics (MPs), forming an EPS-corona that affects the fate of MPs in marine ecosystems. However, the impact of the EPS-corona on the biotoxicity of MPs to marine organisms remains poorly understood. Herein, the effect of the EPS-corona on the toxicity of polystyrene (PS) MPs of different sizes (0.1 and 1 µm) to Skeletonema costatum (S. costatum) was investigated. The preferential adsorption of medium molecule weight (∼55 kDa) proteins onto PS MPs mainly contributed to the EPS-corona formation, decreasing the surface charge negativity of small-sized PS MPs (0.1 µm) by 72.4 %. Nitrogen (N) and oxygen (O) moieties in polysaccharides and proteins were identified as the preferential adsorption sites in the EPS-PS MPs interaction. Density functional theory (DFT) calculations confirmed the nuclear magnetic resonance spectroscopy (NMR) results, revealing that the binding mode between EPS and PS MPs was mainly hydrogen bonding. In addition, EPS-corona increased the cell density of S. costatum by 35.5-36.0 % when exposed to small-sized PS MPs (0.1 µm, 25-50 mg/L). These findings provide new insights into how EPS-corona affects the environmental fate and ecological risks associated with micro- and nano-sized plastics in marine ecosystems.
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  • 文章类型: Journal Article
    活性污泥工艺在现代污水处理厂中起着至关重要的作用。在日常污水处理过程中,产生大量剩余污泥,which,如果管理不当,会对环境和人类健康造成负担。此外,生物聚合物的高度水合的胶体结构限制了脱水的速率和程度,使机械脱水具有挑战性。本研究探讨了微波辐照(MW)与过乙酸(PAA)联用对污泥脱水效率的影响和机理。通过毛细管抽吸时间(CST)和比过滤阻力(SRF)评估污泥脱水效果。检查MW-PAA处理对污泥脱水性能的影响,涉及评估胞外聚合物(EPS)的水平,采用三维激发-发射矩阵(3D-EEM),傅里叶变换红外光谱(FT-IR),和扫描电子显微镜。研究结果表明,最佳脱水性能,SRF和CST分别降低91.22%和84.22%,在以下条件下获得:600W的微波功率,反应时间120s,PAA剂量为0.25g/gMLSS。此外,对MW-PAA处理前后污泥EPS组成和絮凝物形态的变化进行了检查。研究结果表明,微波还可以促进PAA分解为•OH自由基,表明联合MW-PAA治疗具有协同作用。这些相关的研究结果为采用MW-PAA技术处理剩余污泥提供了见解。
    The activated sludge process plays a crucial role in modern wastewater treatment plants. During the treatment of daily sewage, a large amount of residual sludge is generated, which, if improperly managed, can pose burdens on the environment and human health. Additionally, the highly hydrated colloidal structure of biopolymers limits the rate and degree of dewatering, making mechanical dewatering challenging. This study investigates the impact and mechanism of microwave irradiation (MW) in conjunction with peracetic acid (PAA) on the dewatering efficiency of sludge. Sludge dewatering effectiveness was assessed through capillary suction time (CST) and specific resistance to filtration (SRF). Examination of the impact of MW-PAA treatment on sludge dewatering performance involved assessing the levels of extracellular polymeric substances (EPS), employing three-dimensional excitation-emission matrix (3D-EEM), Fourier transform-infrared spectroscopy (FT-IR), and scanning electron microscopy. Findings reveal that optimal dewatering performance, with respective reductions of 91.22% for SRF and 84.22% for CST, was attained under the following conditions: microwave power of 600 W, reaction time of 120 s, and PAA dosage of 0.25 g/g MLSS. Additionally, alterations in both sludge EPS composition and floc morphology pre- and post-MW-PAA treatment underwent examination. The findings demonstrate that microwaves additionally boost the breakdown of PAA into •OH radicals, suggesting a synergistic effect upon combining MW-PAA treatment. These pertinent research findings offer insights into employing MW-PAA technology for residual sludge treatment.
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  • 文章类型: Journal Article
    同时反硝化和产甲烷(SDM)可以有效地消除高浓度的含氮和抗生素废水。重金属和抗生素是导致抗生素抗性基因(ARGs)水平转移的两个关键因素,可以在废水中同时检测到。不幸的是,重金属对SDM和抗生素生物降解的影响尚未完全阐明。在这里,SDM和多种抗生素生物降解的影响,胞外聚合物(EPS)和蛋白质反应机制,对Zn(II)胁迫下的ARG命运进行了综合评估。结果表明,高水平的Zn(II)(≥5mg/L)胁迫显着降低了多种抗生素的降解速率,并抑制了反硝化和甲烷生成。此外,Zn(II)暴露促使蛋白质从微生物中释放到EPS中,EPS与小分子的结合使原始荧光成分猝灭并破坏了蛋白质结构。优势蛋白可以通过几种类型的化学相互作用与Zn(II)和多种抗生素结合,包括金属键和氢键,疏水相互作用,和盐桥,减轻有害物质的毒性。此外,宏基因组测序显示,锌抗性基因(Zn-RGs)的丰度,ARGs(主要是四环素),在Zn(II)胁迫下,可移动遗传元件(MGEs)增加。Mantel测试说明了ARG的mecD,tett,tetB(60)受MGE影响最大。此外,分子网络分析表明,几种MGE可以桥接金属抗性基因(MRGs)和ARGs,促进ARG的水平转移。该研究为SDM系统处理含抗生素废水的环境风险控制提供了理论指导。
    High-strength nitrogen and antibiotics-containing wastewater can be efficiently eliminated by simultaneous denitrification and methanogenesis (SDM). Heavy metals and antibiotics are two critical factors that can lead to horizontal transfer of antibiotic resistance genes (ARGs), which can be simultaneously detected in wastewater. Unfortunately, the impacts of heavy metals on SDM and antibiotic biodegradation have not been fully elucidated. Herein, the effects of SDM and multiple antibiotics biodegradation, extracellular polymeric substances (EPSs) and protein response mechanisms, and ARG fate under Zn(II) stress were comprehensively evaluated. The results indicated that a high level of Zn(II) (≥5 mg/L) stress significantly decreased the degradation rate of multiple antibiotics and suppressed denitrification and methanogenesis. In addition, Zn(II) exposure prompted the liberation of proteins from microbes into the EPSs, and the combination of EPSs with small molecules quenched the original fluorescent components and destroyed the protein structure. The dominant proteins can bind to both Zn(II) and multiple antibiotics through several types of chemical interactions, including metallic and hydrogen bonds, hydrophobic interactions, and salt bridges, relieving the toxicity of harmful substances. Moreover, metagenomic sequencing revealed that the abundance of zinc resistance genes (Zn-RGs), ARGs (mainly tetracyclines), and mobile genetic elements (MGEs) increased under Zn(II) stress. Mantel test illustrated that the ARGs mecD, tetT, and tetB(60) were most affected by MGEs. Moreover, molecular network analysis revealed that several MGEs can bridge metal resistance genes (MRGs) and ARGs, facilitating the horizontal transfer of ARGs. This study provides theoretical guidance for the environmental risk control of antibiotics-containing wastewater treated by an SDM system.
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  • 文章类型: Journal Article
    厌氧氨氧化(anammox)是一种经济有效的技术,但其性能会受到高负载应力的严重抑制。这项研究通过添加一定量的纳米零价铁,在anammox颗粒(AnGS)表面创建了一个创新的富铁结壳层(IEL)。IEL是通过凝胶网络的聚集和铁物种与胞外聚合物(EPS)的结合而形成的,导致沉降能力显著提高,EPS分泌,和血红素含量。宏基因组分析表明,IELAnGS中与氮代谢相关的功能基因显着增加。在高负荷应力下,没有IEL的AnGS的氨去除性能严重下降。相比之下,IELAnGS表现出超过90%的优异的氨去除效率。IEL是AnGS的保护屏障,有效地减轻了强大的剪切力,从而增强它们对高负荷应力的抵抗力。
    Anaerobic ammonia oxidation (anammox) is a cost-effective technology but its performance can be seriously inhibited by high load stress. This study has created an innovative iron-rich encrustation layer (IEL) on the surface of anammox granules (AnGS) through the addition of a certain amount of nano zero-valent iron. The IEL was formed through the aggregation of a gel network and the binding of iron species with extracellular polymeric substances (EPS), resulting in a significant increase in settling ability, EPS secretion, and heme content. Metagenomic analysis indicated a notable rise in the functional genes associated with nitrogen andiron metabolism in IEL AnGS. Under high load stress, the ammonia removal performance of AnGS without IEL severely declined. In contrast, IEL AnGS exhibited excellent ammonia removal efficiency of over 90%. The IEL served as a protective barrier for AnGS, effectively mitigating the strong shear forces, thereby enhancing their resistance to high load stress.
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  • 文章类型: Journal Article
    植物生物刺激剂作为化学肥料的可持续替代品已经受到关注。细胞外聚合物质(EPS),在植物生长促进根际细菌(PGPRs)分泌的化合物中,被假定为减轻非生物胁迫。本研究旨在研究纯化EPS对非生物胁迫下水稻的影响,并分析其作用机制。进行了盆栽实验,以阐明在糖存在下接种从PGPR纯化的EPS对水稻生长的影响。由于所有EPS在应激后显示SPAD的改善,路德维吉肠杆菌,没有表现出更高的PGP生物活性,如植物激素的产生,固氮,和磷的溶解,选择进行进一步分析。在用EPS或水处理24小时后从发芽种子的胚提取的RNA用于转录组分析。RNA-seq分析显示,在水稻种子中鉴定出215个差异表达基因(DEG),包括139个上调基因和76个下调基因。基因本体论(GO)富集分析表明,富集的GO术语主要与ROS清除过程有关,解毒途径,和对氧化应激的反应。例如,编码OsAAO5的基因的表达,已知该基因在解毒氧化应激中起作用,EPS处理增加了两倍。此外,EPS的应用提高了SPAD和茎和根的干重90%,14%,27%,分别,在干旱胁迫下,盐胁迫下SPAD增加59%。这表明细菌EPS在非生物胁迫下改善了植物的生长。根据我们的结果,我们认为从路德维吉肠杆菌中纯化的EPSs可用于开发水稻生物刺激剂。
    Plant biostimulants have received attention as sustainable alternatives to chemical fertilizers. Extracellular polymeric substances (EPSs), among the compounds secreted by plant growth-promoting rhizobacteria (PGPRs), are assumed to alleviate abiotic stress. This study aims to investigate the effect of purified EPSs on rice under abiotic stress and analyze their mechanisms. A pot experiment was conducted to elucidate the effects of inoculating EPSs purified from PGPRs that increase biofilm production in the presence of sugar on rice growth in heat-stress conditions. Since all EPSs showed improvement in SPAD after the stress, Enterobacter ludwigii, which was not characterized as showing higher PGP bioactivities such as phytohormone production, nitrogen fixation, and phosphorus solubilization, was selected for further analysis. RNA extracted from the embryos of germinating seeds at 24 h post-treatment with EPSs or water was used for transcriptome analysis. The RNA-seq analysis revealed 215 differentially expressed genes (DEGs) identified in rice seeds, including 139 up-regulated and 76 down-regulated genes. A gene ontology (GO) enrichment analysis showed that the enriched GO terms are mainly associated with the ROS scavenging processes, detoxification pathways, and response to oxidative stress. For example, the expression of the gene encoding OsAAO5, which is known to function in detoxifying oxidative stress, was two times increased by EPS treatment. Moreover, EPS application improved SPAD and dry weights of shoot and root by 90%, 14%, and 27%, respectively, under drought stress and increased SPAD by 59% under salt stress. It indicates that bacterial EPSs improved plant growth under abiotic stresses. Based on our results, we consider that EPSs purified from Enterobacter ludwigii can be used to develop biostimulants for rice.
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  • 文章类型: Journal Article
    构建了生物电化学上流厌氧污泥层(BE-UASB),并与传统的UASB进行了比较,以研究生物电催化在调节高硫酸盐甲醇废水厌氧处理中的甲烷生成和硫化物生成中的作用(COD/SO42-比值≤2)。BE-UASB的甲烷产率是单一UASB的1.4倍,SO42-去除率稳定在16.7%。生物电催化选择性富集关键功能厌氧菌,刺激细胞外聚合物分泌,特别是有利于电子转移的腐殖酸,从而加速电极的电活性生物膜的发展。甲烷甲烷是直接将甲醇转化为CH4的优势属(35%)。甲烷细菌作为CO2电还原产甲烷的古细菌仅出现在电极上。乙酸杆菌表现出阳极依赖性,通过协同共存为硫酸盐还原细菌(NorankSyntrophobacteraceae和脱硫微生物)提供乙酸盐。本研究证实了BE-UASB调控微生物生态,实现了高硫酸盐甲醇废水的高效去除和能量回收。
    A bioelectrochemical upflow anaerobic sludge blanket (BE-UASB) was constructed and compared with the traditional UASB to investigate the role of bioelectrocatalysis in modulating methanogenesis and sulfidogensis involved within anaerobic treatment of high-sulfate methanolic wastewater (COD/SO42- ratio ≤ 2). Methane production rate for BE-UASB was 1.4 times higher than that of the single UASB, while SO42- removal stabilized at 16.7%. Bioelectrocatalysis selectively enriched key functional anaerobes and stimulated the secretion of extracellular polymeric substances, especially humic acids favoring electron transfer, thereby accelerating the electroactive biofilms development of electrodes. Methanomethylovorans was the dominant genus (35%) to directly convert methanol to CH4. Methanobacterium as CO2 electroreduction methane-producing archaea appeared only on electrodes. Acetobacterium exhibited anode-dependence, which provided acetate for sulfate-reducing bacteria (norank Syntrophobacteraceae and Desulfomicrobium) through synergistic coexistence. This study confirmed that BE-UASB regulated the microbial ecology to achieve efficient removal and energy recovery of high-sulfate methanolic wastewater.
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  • 文章类型: Journal Article
    建立了正四面体模型,以通过高分辨率质谱来刺穿四元成分中溶解有机物(DOM)的分馏。该模型可以立体可视化DOM的分子式,以根据正四面体中的位置显示对每个组件的偏好。随后开发了一种分类方法,将分子式分为与分馏比有关的15类,证明其相对变化与质量峰面积的不确定性一致。以胞外聚合物分层与OrbitrapMS耦合为例,以垃圾渗滤液处理和污水处理厂的7种污泥为例,验证了正四面体模型的实用性,呈现分层污泥絮体中的DOM化学多样性。敏感性分析证明,在四个模型参数的扰动下,分类结果相对稳定。根据正四面体模型的分类结果,多项逻辑回归分析可以进一步帮助识别分子性质对DOM分馏的影响。该模型提供了一种方法,用于评估从固体或半固体成分中顺序提取DOM的特异性,并简化了四元成分分馏系数的复杂数学表达式。
    A regular tetrahedron model was established to pierce the fractionation of dissolved organic matter (DOM) among quaternary components by using high-resolution mass spectrometry. The model can stereoscopically visualize molecular formulas of DOM to show the preference to each component according to the position in a regular tetrahedron. A classification method was subsequently developed to divide molecular formulas into 15 categories related to fractionation ratios, the relative change of which was demonstrated to be convergent with the uncertainty of mass peak area. The practicality of the regular tetrahedron model was verified by seven kinds of sludge from waste leachate treatment and sewage wastewater treatment plants by using stratification of extracellular polymeric substances coupled with Orbitrap MS as an example, presenting the DOM chemodiversity in stratified sludge flocs. Sensitivity analysis proved that classification results were relatively stable with the perturbation of four model parameters. Multinomial logistic regression analysis could further help identify the effect of molecular properties on the fractionation of DOM based on the classification results of the regular tetrahedron model. This model offers a methodology for the assessment of specificity of sequential extraction on DOM from solid or semisolid components and simplifies the complex mathematical expression of fractionation coefficients for quaternary components.
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