Molecular interactions of pesticides at the soil-water interface

High-resolution magic angle spinning (HR-MAS) NMR spectroscopy combined with saturation-transfer double difference (STDD) NMR can be used to analyze the molecular-level interactions of pesticides and whole soils occurring at the soil-water interface. Here 1H HR-MAS STDD NMR has been applied to some common pesticides (trifluralin, acifluorfen, and (4-nitro-3-(trifluoromethyl) phenol) and a pesticide degradation product (1-naphthol). Results indicate that dipolar interactions, H-bonding, hydrophobic associations, and potentially pi-pi interactions are the predominant sorption mechanisms for these molecules at the soil-aqueous interface. It is evident that the physical and chemical characteristics of soil are highly influential in determining the mechanisms of pesticide sorption, as they significantly affect soil conformation. In particular, different binding mechanisms were observed for 1-naphthol in soil swollen using a buffer versus D2O, indicating that the K(oc) alone may not be enough to accurately predict the behavior of a molecule in a real soil environment. Preliminary kinetic-based studies suggest that both the swelling solvent and soil moisture content significantly influence the sequestration of trifluralin. These studies demonstrate that HR-MAS and STDD NMR are powerful and versatile tools which can be applied to expand our knowledge of the mechanistic interactions of agrochemicals at the molecular level.     

Mechanism of interactions between Hg(II) and Demeton S: an NMR study

The chemical fate of organophosphorus pesticides (OPs) has been proven to depend strongly on the chemistry of their aquatic environment. In particular, metal ions (and metal oxide surfaces) have been known to play an important role in the hydrolytic fate of OPs. Various postulates regarding the mechanism of metal-ion-promoted hydrolysis of OPs have been made over the years. However, direct spectroscopic evidence to pinpoint the hydrolytic products and the exact interaction between metal ions and organophosphorus pesticides are still lacking. We report herein the first in-situ study of the interaction between an aqueous solution of Hg(II) and Demeton S using 1H- and 31P NMR spectroscopy. It was found that the interactions between Hg(II), a soft Lewis acid, and Demeton S tend to be a strong function of the aqueous speciation of Hg(II), and the bonding between Hg2+ and Demeton S does not involve the central P=O bond but rather Hg2+ bonds with the two sulfur atoms in the Demeton S side chain and subsequently stabilizes the Demeton S molecule, a phenomenon not previously reported for any metal ion-OP systems studied. On the basis of this study, generalizations regarding the nature of metal ion binding even within a given class of OPs (i.e., phosphorodithioates, phosphorothioates, phosphorothiolates, etc.) should be avoided or only made with extreme caution.     

Thermodynamics of peat-, plant-, and soil-derived humic acid sorption on kaolinite

Humic acids (HAs) form coatings on clays and minerals that can play an important role in nutrient and contaminant migration in soil and water. Humic acid-clay mineral interactions are known to be affected by pH and ionic strength, but little attention has been paid to the effects of temperature. In this paper we report the stoichiometry and thermodynamics of interactions of aqueous HAs (isolated from two peats, two soils and a marine alga with a method that removes lipids) with kaolinite clay, Al2Si2O5(OH)4, at seven temperatures from 5.0 to 35.0 degrees C in 0.05 M NaCl at pH 3.5. All the sorption isotherms exhibit consecutive steps ascribed to HA monolayer and bilayer formation, respectively. Site capacity comparisons suggest different HA molecular conformations on kaolinite. Linearly correlated enthalpy and entropy changes for HA sorption point to the importance of hydration and dehydration in the sorption mechanism.     

Modeling of lithium interference in cadmium biosorption

Biosorption of Cd by the brown seaweed Sargassum polycystum biomass was experimentally investigated and mathematically modeled at different pH and ionic strength levels. From the potentiometric titration of the biomass, three types of functional groups were identified, and the dissociation constant and the numbers of these groups were determined. The carboxyl group (pK(H) 3.70 +/- 0.09) was found to play a major role in binding protons and Cd. The background ion, Li+, could interfere with the uptake of protons and Cd by competition forthe carboxyl sites. Whereas the binding mechanism on the carboxyl group was established as an ion exchange process, the second functional group, phosphonate (pK(H) 5.41 +/- 0.31), most likely bound metals by a complexation reaction. A biosorption model was developed based upon the binding mechanisms and was successfully used for predicting the isotherm and pH edge experiments. In addition, the speciation of the binding sites as a function of the pH was simulated using the developed model in order to visualize the distribution of Cd on the binding sites.     

透明质酸在肿瘤治疗中的应用

自上个世纪30年代透明质酸发现以来,人类对其进行了大量的研究。其在美容、类风湿性关节炎及术后防粘连等方面发挥了很大的功效。目前,根据透明质酸的理化性质,国内外开始着手对其进行肿瘤治疗方面的研究,虽尚没有确切数据予以说明,但还是取得了一定的疗效。本文在前人研究的基础上,综合国内外文献,根据透明质酸与CD44受体的结合作用,通过对其降解,对抗其与CD44受体的结合等方法主要探讨透明质酸在人类肿瘤治疗中的作用。     

Distinct effects of humic acid on transport and retention of TiO2 rutile nanoparticles in saturated sand columns

The distinct effects of humic acid (HA, 0-10 mg L(-1)) on the transport of titanium dioxide (rutile) nanoparticles (nTiO(2)) through saturated sand columns were observed under conditions of environmental relevance (ionic strength 3-200 mM NaCl, pH 5.7 and 9.0). Specifically, the transport of nTiO(2) was dramatically enhanced in the presence of HA at pH 5.7, even at a low HA concentration of 1 mg L(-1). The mobility of nTiO(2) was further increased with greater concentrations of HA. In contrast, this enhancement of the nTiO(2) transportability due to the presence of HA was limited at pH 9.0 because of the negligible adsorption of HA onto nTiO(2), regardless of the concentrations of HA examined in this study. The distinct effects can be explained by the adsorption behaviors of HA to nTiO(2) and sand surfaces and the resulting interactions between nTiO(2) and sand surfaces under different conditions, which resulted in a large variation of the nTiO(2) transport and deposition behaviors at various conditions. In addition, theoretical interaction energy calculations and additional elution experiments indicate that the secondary energy minimum played an important role in controlling the nTiO(2) transport and deposition in porous media observed in this study. Moreover, the interaction energy calculations suggest that at pH 5.7, HA affected nTiO(2) transport by increasing the negative surface charge of nTiO(2) at low HA adsorption densities; whereas, combinations of increased electrostatic and steric interactions due to the presence of HA were the main mechanisms of enhanced transportability of nTiO(2) at high HA adsorption densities. Overall, results from this study suggest that natural organic matter and solution pH are likely key factors that govern the stability and mobility of nTiO(2) in the natural aquatic environment.     

Binding of atrazine to humic substances from soil,peat and coal related to their structure

Partition coefficients for the binding affinities of atrazine to 16 different humic materials were determined by the ultrafiltration HPLC technique. Sources included humic acids (HA), fulvic acids (FA), and combined humic and fulvic fractions (HF) from soil, peat, and coal humic acid. Each of the humic materials was characterized by elemental composition, molecular weight, and composition of main structural fragments determined by 13C solution-state NMR. The magnitude of K(OC) values varied from 87 to 575 L/kg of C, demonstrating relatively low binding affinity of humic substances (HS) for atrazine. On the basis of the measured K(OC) values, the humic materials can be arranged in the following order: coal HA approximately = gray wooded soil HA > chernozemic soil HA and HF > sod-podzolic soil HA approximately = peat HF > sod-podzolic soil FA > peat dissolved organic matter. The magnitude of the K(OC) values correlated strongly with the percentage of aromatic carbon in HS samples (r = 0.91). The hydrophobic binding was hypothesized as the key interaction underlying the binding of atrazine to HS.     

Nickel sequestration in a kaolinite-humic acid complex

Incorporation of first row transition metals into stable surface precipitates can play an important role in reducing the bioavailability of these metals in neutral and alkaline soils. Organic coatings may interfere with this sorption mechanism by changing the surface characteristics and by masking the mineral surface from metal sorptives. In this study, kinetic sorption and desorption experiments were combined with extended X-ray absorption fine structure (EXAFS) spectroscopy to elucidate the effect of humic acid (HA) coatings on the formation and stabilization of nickel precipitates at the kaolinite-water interface. Initial Ni uptake (pH 7.5, [Ni]i = 3 mM, and I = 0.02 M NaNO3) increased with greater amounts of HA coated onto the kaolinite surface. Ni uptake continued over an extended period of time without reaching an apparent equilibrium. EXAFS analysis of the Ni sorption complex structures formed over time (up to 7 months) revealed the formation of a Ni-Al layered double hydroxide (LDH) precipitate at the kaolinite surface in the absence of HA. HA alone formed an inner-sphere complex with Ni (with 2 carbon atoms at an average radial distance of 2.85 A). A Ni-Al LDH precipitate phase was formed at the kaolinite surface in the presence of a 1 wt % HA coating. However, with 5 wt % HA coated at the kaolinite surface, the formation of a surface precipitate was slowed significantly, and the precipitate formed was similar in structure to Ni(OH)2(s). The Ni(OH)2 precipitate was not resistant to proton dissolution, while the Ni-Al LDH precipitate was. These results augment earlier findings that the incorporation of Ni and other first row transition metals into stable surface precipitates is an important sequestration pathway for toxic metals in the environment, despite the presence of ubiquitous coating materials such as humic acids.     

Effect of pH on the in vitro interactions between bile acids and pectin

 The interactions between pectin and bile acids (BA) were investigated after in vitro incubation at 37°C in the pH range of 5.0 – 8.0, separation of the non-absorbed BA using membrane filtration and their estimation by HPLC and HPTLC. In general, the interactions increased with the degree of esterification and the molecular weight (viscosity) of the pectin. They were reduced if the carboxyl groups in the polysaccharide molecules are arranged in a blockwise manner and in the presence of Ca²⁺. The highest interactions were found for unconjugated BA, followed by the corresponding glycine-conjugates. Taurine-conjugates showed the lowest interaction with pectin. Chenodeoxycholic acid and deoxycholic acid reacted more effectively than the trihydroxy bile acid cholic acid in the in vitro experiments. The results show that structural parameters of both BA and pectin play an important role in the extent of their interaction. Received: 28 November 1996 / Revised version: 20 January 1997     

Effect of pH on the in vitro interactions between bile acids and pectin

 The interactions between pectin and bile acids (BA) were investigated after in vitro incubation at 37°C in the pH range of 5.0 – 8.0, separation of the non-absorbed BA using membrane filtration and their estimation by HPLC and HPTLC. In general, the interactions increased with the degree of esterification and the molecular weight (viscosity) of the pectin. They were reduced if the carboxyl groups in the polysaccharide molecules are arranged in a blockwise manner and in the presence of Ca²⁺. The highest interactions were found for unconjugated BA, followed by the corresponding glycine-conjugates. Taurine-conjugates showed the lowest interaction with pectin. Chenodeoxycholic acid and deoxycholic acid reacted more effectively than the trihydroxy bile acid cholic acid in the in vitro experiments. The results show that structural parameters of both BA and pectin play an important role in the extent of their interaction. Received: 28 November 1996 / Revised version: 20 January 1997     

Impact of de-ashing humic Acid and humin on organic matter structural properties and sorption mechanisms of phenanthrene

Organic matter-mineral interactions greatly affect the fate of hydrophobic organic compounds (HOCs) in the environment. In the present study, the impact of organic matter-mineral interaction on sorption of phenanthrene (PHE) by the original and de-ashed humic acids (HAs) and humin (HM) was examined. After de-ashing treatment, the overall polarity of organic matter in HAs and HM consistently decreased. Differently, the surface polarity of HAs increased but that of HM decreased. No correlation between K(oc) values of PHE by all tested sorbents and their bulk polarity was observed due to inaccessibility of a portion of interior sorption domains. The inaccessibility of interior sorption domains in HAs and HM was partly due to the crystalline structure in organic matter as indicated by differential scanning calorimetric (DSC) and 13C NMR data and the interference from minerals. A good correlation between surface polarity of the original and de-ashed HAs and HMs and their K(oc) values for PHE indicated its importance in HOC sorption. Dissimilar changes in surface polarity of HAs and HM after de-ashing treatment can be ascribed to the distinct interactions between organic matter and minerals. The solid-state 13C NMR, XPS, and elemental composition data of all tested sorbents revealed that a larger fraction of O atoms in HAs were involved in organic matter-mineral interaction as compared to HM. Results of this work highlight the importance of soil organic matter (SOM)-mineral interactions, surface polarity, and microscaled domain arrangement of SOM in HOC sorption.     

Evidence for nonlinear binding of PAHs to dissolved humic acids

Binding of pyrene, fluoranthene, and phenanthrene to dissolved humic acids (HA) was determined by the fluorescence quenching (FQ) and complexation-flocculation (CF) methods. Determinations by the CF method, using varying contaminant concentrations and a constant HA concentration, yielded nonlinear Freundlich-type isotherms (n = 0.65-0.84). Experiments using both the CF and the FQ methods with varying HA concentrations and a constant contaminant concentration yielded curved "Stern-Volmer"-type plots that also indicate nonlinear binding. These findings suggest that linear partitioning or site complexation in the presence of excess available sites cannot fully describe the interactions of hydrophobic compounds with dissolved humic material. Site-specific hydrophobic interactions at limited interior or external molecular surfaces may be considered.     

Quantitative evaluation of noncovalent interactions between glyphosate and dissolved humic substances by NMR spectroscopy

Interactions of glyphosate (N-phosphonomethylglycine) herbicide (GLY) with soluble fulvic acids (FAs) and humic acids (HAs) at pH 5.2 and 7 were studied by (1)H and (31)P NMR spectroscopy. Increasing concentrations of soluble humic matter determined broadening and chemical shift drifts of proton and phosphorus GLY signals, thereby indicating the occurrence of weak interactions between GLY and humic superstructures. Binding was larger for FAs and pH 5.2 than for HAs and pH 7, thus suggesting formation of hydrogen bonds between GLY carboxyl and phosphonate groups and protonated oxygen functions in humic matter. Changes in relaxation and correlation times of (1)H and (31)P signals and saturation transfer difference NMR experiments confirmed the noncovalent nature of GLY-humic interactions. Diffusion-ordered NMR spectra allowed calculation of the glyphosate fraction bound to humic superstructures and association constants (K(a)) and Gibbs free energies of transfer for GLY-humic complex formation at both pH values. These values showed that noncovalent interactions occurred most effectively with FAs and at pH 5.2. Our findings indicated that glyphosate may spontaneously and significantly bind to soluble humic matter by noncovalent interactions at slightly acidic pH and, thus, potentially pollute natural water bodies by moving through soil profiles in complexes with dissolved humus.     

大米蛋白与米渣蛋白对镉结合能力的对比研究

目的:比较大米蛋白和米渣蛋白对镉的结合能力,探究蛋白对镉的结合机理。方法:通过蛋白对镉的结合试验、镉的脱除试验,比较两种蛋白对镉的结合能力,并用粒径仪、圆二色光谱仪(CD)、红外光谱仪(FTIR)、扫描电子显微镜(SEM)、X射线光电子能谱分析仪(XPS)等仪器分析蛋白与镉的结合机理。通过掩蔽蛋白羧基、氧化巯基后再对镉结合的试验,验证蛋白的羧基及巯基对镉结合的重要性。结果:米渣蛋白对镉的最大结合量为12.08 mg/g,被结合的镉120 min后脱除率达到最大,大米蛋白对镉的最大结合量为8.85 mg/g,被结合的镉60 min后脱除率达到最大。掩蔽蛋白羧基后,两种蛋白对镉的结合量较羧基掩蔽前均下降18%;氧化巯基后,大米蛋白和米渣蛋白较巯基被氧化前对镉的结合量分别下降40%和50%。SEM观察出,蛋白结合镉后发生聚集现象。结论:与大米蛋白相比,米渣蛋白对镉的结合能力更强。此外,两种蛋白对镉的结合不仅是简单的物理吸附作用,蛋白中的巯基和羧基对镉的结合也具有重要作用。     

多肽螯合金属离子产物的研究进展

机体内存在的金属离子发挥着至关重要的作用,金属元素的缺乏会导致不良症状的产生。目前市场上的金属元素补充剂价格较高或对胃肠道具有一定的刺激性,因此急需高效廉价的新型补充剂来应对此类现状。其中,由蛋白质水解产生的具有金属离子螯合性的多肽具有深远的研究意义。多肽金属离子螯合物是蛋白质水解产生的小分子多肽的羧基、磷酸基团、酰胺基团以及氨基酸等与金属离子通过配位共价结合或者吸附结合后产生的活性产物,具有生物利用率高、安全性高、生物活性高等优点。本文综述了国内外金属离子螯合肽的制备、肽金属螯合物的结构表征、酶解物的分离纯化、螯合肽的生物利用率及螯合机制的相关研究,并对目前现存在的问题进行总结,以期对矿物质元素补充剂的研究与开发提供新思路。     

Monitoring the effect of three humic acids on a model membrane system using 31P NMR

The sorption of three humic acids to 1,2-dipalmitoyl-sn-glycero-3-phosphocholine multilamellar vesicle model membrane systems was studied by phosphorus nuclear magnetic resonance (31P NMR). The effects of temperature and pH were investigated. The gel --> bilayer transition did not appear to be affected by any of the humic acids at pH 7; however, all three humic acids induced a perturbation to this transition and to the bilayer structure at pH 4. On the basis of the findings from this and other work, a conceptual adsorption/absorption model for the sorption of humic acid (HA) to biomembranes has been put forward. The model requires an initial adsorption step initiated at an acidic pH by hydrogen bridging and electrostatic interactions between the functional groups of the HAs and the head groups of the phospholipids. Once the HA material is adsorbed, its hydrophobic domains can further seek a more thermodynamically favorable environment within the bilayer using hydrophobic interactions. These interactions lead to the HA being absorbed into the membrane, which subsequently induces the observed perturbation by disturbing the ordered packing of the phospholipid tail groups. This model is also related to other humic substances/biomembrane observations in the literature.     

Mechanisms of humic acids degradation by white rot fungi explored using 1H NMR spectroscopy and FTICR mass spectrometry

Enzymatic activities involved in decay processes of natural aromatic macromolecules, such as humic acids (HA) and lignin by white rot fungi, have been widely investigated. However, the physical and chemical analysis of degradation products of these materials has not been intensively explored. Fourier transform cyclotron resonance mass spectrometry (FTICR MS) and 1H NMR as well as CHNOS and size exclusion chromatography were employed to study the mechanisms of HA degradation by Trametes sp. M23 and Phanerochaete sp. Y6. Size exclusion chromatography analyses demonstrate and provide evidence for HA breakdown into low MW compounds. The 1H NMR analysis revealed oxidation, a decrease in the aromatic content, and an indication of demethylation of the HA during biodegradation. Evidence for oxidation was also obtained using CHNOS. Analysis of FTICR MS results using a new software program developed by our group (David Mass Sort) revealed consecutive series of masses suggesting biochemical degradation trends such as oxidation, aromatic cleavage, and demethylation. These results are in agreement with the 1H NMR analysis and with the suggested role of the ligninolytic system leading to HA degradation.     

Enhanced pH and thermal stabilities of invertase immobilized on montmorillonite K-10

Invertase was adsorbed onto micro-porous acid-activated montmorillonite clay (K-10) by two procedures, namely adsorption and covalent binding. The immobilized enzymes were characterized by XRD, surface area measurements and ²⁷Al NMR. XRD measurements revealed an expansion of clay layers due to immobilization which suggests that intercalation had taken place. Surface area measurements also support this observation. ²⁷Al NMR showed that interaction of enzyme with tetrahedral and octahedral Al changes with the immobilization procedure. Sucrose hydrolysis was performed in a batch reactor. The immobilized enzymes showed enhanced pH and thermal stabilities. Optimum pH and temperature were found to increase upon immobilization. The effectiveness factor (η) and Michaelis constant (K_m) suggest that diffusional resistances play a major role in the reaction. The immobilized invertase could be stored in buffer of pH 5 and 6 at 5 °C without any significant loss in activity for 20 days.     

低酯果胶-咖啡酸交联物的构成机理及与抗氧化活性的构效关系

本研究采用绿色安全的漆酶催化法制备柑橘低酯果胶（citrus low-methoxyl pectin,CLP）和咖啡酸（caffeic acid,CaA）交联物,解析交联物的关键组成、光谱特征、核磁共振图谱特征以及体外抗氧化活性,以明确其交联机理、分子间键结模式,并归纳其贡献抗氧化活性的结构因素。化学成分组成与离子色谱分析结果显示,CLP含有51.99%（摩尔百分比,下同）半乳糖醛酸（galacturonic acid,GalA）、30.95%半乳糖（galactose,Gal）、7.44%鼠李糖（rhamnose,Rha）和9.02%葡萄糖（glucose,Glc）及微量总酚（1.20 mg/g）,酯化度为38.33%,CLP主要分子链段由同型半乳糖醛酸聚糖（homogalacturonan,HG）和第一型鼠李糖半乳糖醛酸聚糖（rhamnogalacturonan,RG-I）所组成,其中RG-I的中性糖侧链为聚半乳糖。CLP的重均分子质量（mw）为226.1 kDa。相较于CLP,CLP-CaA交联物的酯化度和总酚含量分别显著提高到46.93%和9.64 mg/g（P＜0.05）;mw增加到271.4 kDa;紫外吸收光谱和傅里叶变换红外光谱都呈现与CaA有关的特征吸收峰（分别在288 nm和1 519 cm－1）。700 MHz 1H和176 MHz 13C核磁共振图谱清楚地显示了CLP的α-D-GalA、β-D-Gal和部分α-L-Rha糖基的共振尖峰,可推断其主要结构;而交联物CLP-CaA中CLP的糖基碳都有新共振尖峰,位移最大的是α-D-GalA的C1和C4,伴随C6羧基碳的尖峰位移到δC 175.12,且氢谱上出现CaA的特征尖峰（δH 6～8）。综上可知,CLP-CaA的交联机理为：漆酶催化CaA作用于CLP的α-D-GalA糖基C6羧基上形成酯基,且1分子CaA与1分子自由羧基结合。此交联作用促使CLP-CaA具有优越且多元的体外抗氧化活性,对1,1-二苯基-2-三硝基苯肼自由基、2,2’-联氮-双-3-乙基苯并噻唑啉-6-磺酸阳离子自由基、·OH和O2－·的半抑制浓度（half maximal inhibitory concentration,IC50）分别为0.82、2.47、0.92 mg/mL和0.72 mg/mL。本研究发现了果胶与CaA的交联机理及其结构对抗氧化活性的贡献,可为新型改良果胶在食品工业中的应用提供理论参考。     

Localization of CD9 in pig oocytes and its effects on sperm-egg interaction

CD9 is a cell surface protein that participates in many cellular processes, such as cell adhesion. Fertilization involves sperm and oocyte interactions including sperm binding to oocytes and sperm-oocyte fusion. Thus CD9 may play an essential role during fertilization in mammals. The present study was conducted to examine whether CD9 is present in porcine gametes and whether it participates in the regulation of sperm-oocyte interactions. The presence of CD9 in ovarian tissues, oocytes and spermatozoa was examined by immunohistochemistry, immunofluorescence and immunoblotting. Sperm binding and penetration of oocytes treated with CD9 antibody were examined by in vitro fertilization. The results showed that CD9 was present on the plasma membrane of oocytes at different developmental stages. A 24 kDa protein was found in oocytes during in vitro maturation by immunoblotting and its quantity was significantly (P < 0.001) increased as oocytes underwent maturation and reached the highest level after the oocytes had been cultured for 44 h. No positive CD9 staining was found in the spermatozoa. Both sperm binding to ooplasma and sperm penetration into oocytes were significantly (P < 0.01) reduced in anti-CD9 antibody-treated oocytes (1.2 +/- 0.2 per oocyte and 16.6% respectively) as compared with oocytes in the controls (2.5 +/- 0.4 per oocyte and 70.3% respectively). These results indicated that CD9 is expressed in pig oocytes during early growth and meiotic maturation and that it participates in sperm-oocyte interactions during fertilization.