海洋微藻产生的不同粒级溶解有机物质的三维荧光光谱

通过过滤(0.7μm)和切向超滤(100kDa、10kDa、1kDa)技术将实验室内培养的赤潮异弯藻和中肋骨条藻藻液分离成100kDa-0.7μm、10～100kDa、1～10kDa的胶体浓缩液和＜1kDa的超滤液,利用激发-发射矩阵荧光光谱技术测定了不同粒级溶液中有机物的荧光性质,分析了三维荧光光谱图中的荧光峰位置、数量及荧光强度的变化情况.结果表明,两种微藻产生的类腐殖质荧光物质主要是＜1kDa的小分子物质.赤潮异湾藻产生的类蛋白荧光物质中,15%是100kDa-0.7μm的大胶体,8%是10～100kDa的中胶体,而大部分是＜1kDa的小分子物质.赤潮异弯藻各级超滤截留液的类蛋白荧光峰的发射波长与过滤液相比,发生蓝移,而超滤液的则发生红移,表明赤潮异湾藻产生的类蛋白荧光物质随着粒径的增大极性减弱.     

Fluorescence excitation-emission matrix (EEM) spectroscopy for rapid identification and quality evaluation of cell culture media components

The application of fluorescence excitation-emission matrix (EEM) spectroscopy to the quantitative analysis of complex, aqueous solutions of cell culture media components was investigated. These components, yeastolate, phytone, recombinant human insulin, eRDF basal medium, and four different chemically defined (CD) media, are used for the formulation of basal and feed media employed in the production of recombinant proteins using a Chinese Hamster Ovary (CHO) cell based process. The comprehensive analysis (either identification or quality assessment) of these materials using chromatographic methods is time consuming and expensive and is not suitable for high-throughput quality control. The use of EEM in conjunction with multiway chemometric methods provided a rapid, nondestructive analytical method suitable for the screening of large numbers of samples. Here we used multiway robust principal component analysis (MROBPCA) in conjunction with n-way partial least squares discriminant analysis (NPLS-DA) to develop a robust routine for both the identification and quality evaluation of these important cell culture materials. These methods are applicable to a wide range of complex mixtures because they do not rely on any predetermined compositional or property information, thus making them potentially very useful for sample handling, tracking, and quality assessment in biopharmaceutical industries.     

海洋浮游植物生长过程中溶解有机物质的三维荧光光谱研究

利用三维激发-发射矩阵荧光光谱技术,监测了赤潮异弯藻和中肋骨条藻培养过程中产生的溶解有机物,分析了三维荧光光谱图中的荧光峰位置、数量及荧光强度的变化情况.结果表明,微藻生长过程中会产生类蛋白和类腐殖质两类荧光有机物质,这两类有机物的荧光峰的位置及荧光强度有较大差异.在不同生长期,两类荧光有机物的产生机制不同.在指数生长期,两类有机物的荧光强度与藻密度成正相关,说明浮游植物释放了一定量的荧光物质;在平稳期和衰亡期,两类有机物的荧光强度迅速增加,这可能由于衰老、死亡藻细胞的破碎释放出大量的荧光有机物质所致,此外细菌对非荧光有机物进一步降解,也可能是产生该现象的一个原因.     

Excitation-emission matrix fluorescence spectroscopy for natural organic matter characterization: a quantitative evaluation of calibration and spectral correction procedures

The influence of different data collection procedures and of wavelength-dependent instrumental biases on fluorescence excitation-emission matrix (EEM) spectral analysis of aqueous organic matter samples was investigated. Particular attention was given to fluorescence contours (spectral shape) and peak fluorescence intensities. Instrumental bias was evaluated by independently applying excitation and emission correction factors to the raw excitation and emission data, respectively. The peak fluorescence intensities of representative natural organic matter and tryptophan were significantly influenced by the application of excitation and emission spectral correction factors and by the manner in which the raw data was collected. Humification and fluorescence indices were also influenced by emission correction factors but were independent of reference (excitation) intensity normalization or correction. EEM surface contours were dependent on normalization of the fluorescence intensity to the reference intensity but were not influenced by either excitation or emission spectral correction factors. Authors should be explicit in how excitation and emission spectral correction procedures are implemented in their investigations, which will help to facilitate intra-laboratory comparisons and data sharing.     

Waveband selection for NIR spectroscopy analysis of soil organic matter based on SG smoothing and MWPLS methods

Savitzky-Golay (SG) smoothing and moving window partial least square (MWPLS) methods were applied to the model optimization and the waveband selection for near-infrared (NIR) spectroscopy analysis of soil organic matter. The optimal single wavelength prediction bias (OSWPB) was used to evaluate the similarity of calibration set and prediction set, and a new division method for calibration set and prediction set was proposed. SG smoothing modes were expanded to 540 kinds. The specific computer algorithm platforms for optimization of SG smoothing mode combined with PLS factor and for MWPLS method with changeable parameters were built up. The optimal waveband for soil organic matter was 1926-2032 nm, the optimal smoothing mode was the 2nd order derivative, 6th degree polynomial, 45 smoothing points, the PLS factor, RMSEP and R_P were 8, 0.260 (%) and 0.877 respectively. The prediction effect was obviously better than that in the whole spectral collecting region. To get stable results, all the optimization processes were based on the average prediction effect on 50 different divisions of calibration set and prediction set.     

Fast quantification of humic substances and organic matter by direct analysis of sediments using DRIFT spectroscopy

A simple method based on diffuse reflectance coupled with infrared Fourier transform spectroscopy (DRIFTS) has been developed for the quantification and the characterization of sedimentary (or soil, peat, etc.) humic substances. Under optimized conditions, the quantification of humic substances or total organic matter is possible with DRIFTS at a frequency of 2930 cm(-1) using whole dry sediment samples. A study of the operational parameters that affect the DRIFTS signal shows the importance of normalizing analysis conditions, especially the diffuse reflectance accessory alignment, the particle size and compaction, and the homogeneity of the powdered samples, to obtain reproducible quantitative analyses. The quantification of total humic substances by DRIFTS correlates well with the concentrations determined using classical extraction methods. DRIFTS analysis requires only a few minutes instead of tedious extractions of humic substances. Moreover, the distribution of total organic matter and of fulvic acids, humic acids, and humin can also be obtained. Analysis of natural samples indicates that a calibration using humic material representative of the studied area provides the most accurate quantification. The fast screening of organic matter fractions by DRIFTS on intact natural samples provides useful quantitative and qualitative information that can be used in environmental or monitoring studies.     

Impact of low molecular weight organic acids and dissolved organic matter on sorption and mobility of isoproturon in two soils

Isoproturon is a selective herbicide belonging to the phenylurea family and widely used for pre- and post-emergence control of annual weeds. Soil amendments (e.g. organic compounds or dissolved organic matter) may affect environmental behavior and bioavailability of pesticides. However, whether the physiochemical process of isoproturon in soils is affected by organic amendments and how it is affected in different soil types are unknown. To evaluate the impact of low molecular weight organic acids (LMWOA) and dissolved organic matter (DOM) on sorption/desorption and mobility of isoproturon in soils, comprehensive analyses were performed using two distinct soil types (Eutric gleysols and Hap udic cambisols). Our analysis revealed that adsorption of isoproturon in Eutric gleysols was depressed, and desorption and mobility of isoproturon were promoted in the presence of DOM and LMWOA. However, the opposite result was observed with Hap udic cambisols, suggesting that the soil type affected predominantly the physiochemical process. We also characterized differential components of the soils using three-dimensional excitation-emission matrix (EEM) fluorescence spectroscopy and Fourier transform infrared (FT-IR) spectroscopy and show that the two soils displayed different intensity of absorption bands for several functional groups.     

Selective determination of aromatic sulfonates in landfill leachates and groundwater using microbore liquid chromatography coupled with mass spectrometry

Aromatic sulfonates (AS) are large-volume chemicals used in many technical processes of, for instance, the textile industry or construction. A LC/MS method for the selective determination of AS in environmental samples, based on a single-quadrupole MS, was developed and validated. The central point of this technique is the use of the compound-specific fragment ion SO3.- as marker for aromatic sulfonates. This negatively charged SO3 radical, together with the fact that AS undergo loss of SO2, allows screening for AS in complex matrixes, even in the presence of sulfate anions. Calibration curves generated from LC/MS data showed good linearity over 3 orders of magnitude, with an absolute limit of detection of approximately 1 ng. The relative standard deviation for mean areas obtained from reconstructed ion chromatograms ranged from 2.9 to 8.6%. Unlike UV detection, this LC/MS method gives similar response for both naphthalene- and benzene-sulfonates. The method presented was successfully applied to landfill leachates and groundwater, downstream of a landfill. Furthermore, this technique allowed identification of an unknown AS found in drain samples.     

Nonequilibrium solid-phase microextraction for determination of the freely dissolved concentration of hydrophobic organic compounds: matrix effects and limitations

Solid-phase microextraction (SPME) has recently been applied to measure the freely dissolved concentration, as opposed to the total concentration, of hydrophobic substances in aqueous solutions. This requires that only the freely dissolved analytes contribute to the concentration in the SPME fiber coating. However, for nonequilibrium SPME the sorbed analytes that diffuse into the unstirred water layer (UWL) adjacent to the SPME fiber can desorb from the matrix and contribute to the flux into the fiber. These processes were described as a model. Experimentally, an equilibrated and disconnected headspace was used as a reference for the freely dissolved concentration. The expected contribution of desorbed analytes to the uptake flux was measured for PCB no. 52 in a protein-rich solution, while it was not measured in a matrix containing artificial soil. The latter was possibly due to slow desorption of the analyte from the artificial soil. On the basis of the present study, a contribution of desorbed analytes to the uptake flux is expected only if(1) the rate-limiting step of the uptake process is diffusion through the UWL, (2) the concentration of the sorbed analyte is high, and (3) desorption from the matrix is fast.     

Fluorescence lifetime imaging and spectroscopy as tools for nondestructive analysis of works of art

A system for advanced fluorescence investigation of works of art has been assembled and integrated in a characterization procedure that allows one to localize and identify organic compounds that are present in artworks. At the beginning of the investigation, fluorescence lifetime imaging and spectroscopy address a selective microsampling of the artwork. Then analytical measurements of microsamples identify the chemical composition of the materials under investigation. Finally, on the basis of fluorescence lifetime and amplitude maps, analytical data are extended to the whole artwork. In such a way, information on the spatial distribution of organic materials can be inferred. These concepts have been successfully applied in an extensive campaign for analysis of Renaissance fresco paintings in Castiglione Olona, Italy. Residue of various types of glue and stucco left from a restoration carried out in the early 1970s was localized and classified. Insight into the technique used by the painter to make gilded reliefs was also obtained.     

Molecular size distribution of dissolved organic matter in water of the Pearl River and trihalomethane formation characteristics with chlorine and chlorine dioxide treatments

River water sample was collected from Guangzhou section of the Pearl River to investigate soluble organic fractions and formation of trihalomethane (THMs) after chlorine and chlorine dioxide treatments. The water sample was passed through Amicon YC-05, YM-1, YM-3, YM-10, YM-30, YM-100 and ZM-500 series membranes after a pre-treatment. The molecular weight distribution and the specific ultra-violet absorbance (SUVA(254)) of each fraction obtained from membrane were analyzed, and these fractions were further disinfected with chlorine and chlorine dioxide. The results showed that reverse osmosis (RO) fraction contained mainly dissolved organic matter (DOM) from the water sample, suggesting that the water has been highly contaminated by anthropogenic activities. Meanwhile, the THMs concentration and SUVA(254) increased gradually as the molecular weight of the obtained fractions reduced, indicating that the low molecular weight DOM was the major THMs precursor in the disinfection process with chlorine and chlorine dioxide. The results suggest that THMs in source water of Pearl River could be effectively reduced when pollution of human activity is greatly controlled. Between the two disinfection processes tested, chlorine dioxide produced less THMs than chlorine in this study.     

Study of double bond equivalents and the numbers of carbon and oxygen atom distribution of dissolved organic matter with negative-mode FT-ICR MS

A strong linear relationship was observed between the average double bond equivalence (DBE) and the ratio of carbon to oxygen atoms in oxygenated compounds of dissolved organic matter (DOM). Data were acquired by a Fourier transform ion cyclotron resonance mass spectrometer (FT-ICR MS), equipped with a negative-mode electrospray ionization source. The slope and y-intercepts extracted from the linear relationship can be used to compare DOM samples originating from different locations. Significant differences in these parameters were observed between inland riverine and offshore coastal DOM samples. Offshore coastal DOM molecules underwent a change of one DBE for each removal or addition of two oxygen atoms. This suggested the existence of multiple carboxyl groups, each of which contains a double bond and two oxygen atoms. Inland riverine samples exhibited a change of ~1.5 DBE following the addition or removal of two oxygen atoms. This extra change in DBE was attributed to cyclic structures or unsaturated chemical bonds. The DBE value with maximum relative abundance and the minimum DBE value for each class of oxygenated compounds showed that approximately two oxygen atoms contributed to a unity change in DBE. The qualitative analyses given here are in a good agreement with results obtained from analyses using orthogonal analytical techniques. This study demonstrates that DBE and the carbon number distribution, observed by high resolution mass spectrometry, can be valuable in elucidating and comparing structural features of oxygenated molecules of DOM.     

Seasonal variations of chemical and physical characteristics of dissolved organic matter and trihalomethane precursors in a reservoir: a case study

Dissolved organic matter (DOM) and its potential to form disinfection by-products (DBPs) during water treatment are of great public health concern. Understanding the seasonal changes in DOM composition and their reactivity in DBP formation could lead to a better treatment of drinking water and a more consistent water quality. DOM from the East-Lake, a reservoir in the south-China, was fractionated and characterized by XAD resin adsorption (RA) and ultrafiltration (UF) techniques during different seasons within a year. The properties of chemical fractions (isolated by RA) appeared more stable than those of physical fractions (separated by UF) throughout the sampling period. The relative contribution of each chemical fraction to the total dissolved organic carbon (DOC), UV(254) absorbance and trihalomethane formation potential (THMFP) remained relatively constant across the sampling period. However, the physical (molecular weight) fractions of the DOM exhibited large seasonal changes in UV(254) and THMFP. Compared to the parameter of DOC, the THMFP and specific THMFP (STHMFP) of either chemical or physical fractions were more variable. In terms of DOC concentration, the hydrophobic acids (HoA) and hydrophilic matter (HiM) dominated in the DOM in most of the seasons; while the components with molecular weight of 10-30 kDa and less than 1 kDa were the predominant physical fractions.     

Chlorophyll biomass in the global oceans: airborne lidar retrieval using fluorescence of both chlorophyll and chromophoric dissolved organic matter

For three decades airborne laser-induced fluorescence has demonstrated value for chlorophyll biomass retrieval in wide-area oceanic field experiments, satellite validation, and algorithm development. A new chlorophyll biomass retrieval theory is developed using laser-induced and water Raman normalized fluorescence of both (a) chlorophyll and (b) chromophoric dissolved organic matter (CDOM). This airborne lidar retrieval theory is then independently confirmed by chlorophyll biomass obtained from concurrent (1) ship-cruise retrievals, (2) satellite inherent optical property (IOP) biomass retrievals, and (3) satellite standard band-ratio chlorophyll biomass retrievals. The new airborne lidar chlorophyll and CDOM fluorescence-based chlorophyll biomass retrieval is found to be more robust than prior lidar methods that used chlorophyll fluorescence only. Future research is recommended to further explain the underlying influence of CDOM on chlorophyll production.     

A matrix formulation for the dynamic analysis of spatial mechanisms using point coordinates and velocity transformation

Summary. This paper presents a matrix formulation for the dynamic analysis of spatial mechanisms with common types of kinematic joints. The formulation is derived in two steps. Initially an equivalent constrained system of particles that replaces the rigid bodies is constructed and used to define the configuration of the mechanical system. This results in a simple and straightforward procedure for generating the equations of motion in terms of the rectangular Cartesian coordinates of the particles without introducing any rotational coordinates. The equations of motion are then derived in terms of relative joint coordinates through the use of a velocity transformation matrix. The velocity transformation matrix relates the relative joint velocities to the Cartesian velocities. For the open loop case, this process automatically eliminates all of the non-working constraint forces and leads to an efficient integration of the equations of motion. For the closed loop case, suitable joints should be cut and few cut-joints constraint equations should be included for each closed loop. An example is used to demonstrate the generality and efficiency of the proposed method.     

Metal sorption on soils as affected by the dissolved organic matter in sewage sludge and the relative calculation of sewage sludge application

To evaluate the influences of sewage sludge-derived organic matters on metal sorption and on the resultant sludge loading estimates, a batch experiment was conducted to compare the sorption of Ni, Cu and Pb in sewage sludge filtrates (1:20 sewage sludge to water) on eight soils and the adsorption of metals in a reference solution which had the same matrix as the sewage sludge filtrate except dissolved organic material (henceforth referred to as reference solution). Metal sorption could be well fitted by linear isotherm and the dissolved organic matter in sludge significantly depressed the sorption (p<0.01). The main factor controlling sorption of Ni on different soils was dominated by soil cation exchange capacity (CEC) and sorption of Cu and Pb was by soil organic matter (SOM). The parameters obtained from the sorption isotherm equations were then used to estimate sludge loadings into the soils. When the sorption parameters derived from the reference solution were used for calculation, that is the effect of dissolved organic matter was not considered, the calculated safe application rates are approximately 47.8, 51.4, 34.2, 31.3, 21.7, 46.3, 187.1 and 27.6 t-sludge/ha for the Beijing, Jiangxi, Xiamen, Jilin, Guangdong, Wuhan, Gansu and Xinjiang soils, respectively. However, when the sorption parameters derived from the dissolved organo-metallic complexes are used for calculation, the corresponding application rates are reduced to approximately 6.0, 3.4, 1.9, 10.0, 6.3, 3.6, 7.3 and 3.5 t-sludge/ha, respectively. By this study we can get a conclusion that the effect of sewage sludge derived dissolved organic matter on heavy metal sorption and soil properties should be considered in the course of regulating the safe application rates of sewage sludge to soil.     

Thermal desorption/pyrolysis coupled with photoionization time-of-flight mass spectrometry for the analysis of molecular organic compounds and oligomeric and polymeric fractions in urban particulate matter

Atmospheric aerosols are subject to be responsible for human health effects. In this context, besides mass and number concentration of particles, their chemical composition has gained interest recently. However, knowledge about the organic content of particulate matter is still relatively scarce; i.e., only 10-40% of compounds present in the aerosol are as yet identified. By means of a newly developed measurement technique, thermal desorption/photoionization time-of-flight mass spectrometry (TOFMS), organic species evolved from urban aerosol samples collected at Augsburg, Germany, are analyzed. Thereby, compounds desorbed according to a temperature protocol following procedures for OC/EC analysis (120, 250, and 340 degrees C as desorbing temperatures) are ionized by soft, fragmentationless resonance multiphoton ionization (REMPI) and single photon ionization (SPI), respectively. With REMPI-TOFMS, a large variety of PAH is detectable. A comprehensive analysis is enabled by adding SPI-TOFMS, which gives access to aliphatic and carbonylic hydrocarbons as well as alkanoic acids and esters. Analysis of the data showed a high abundance of phenol and guiacol as well as retene, which are known markers for wood combustion. Similar patterns were found with ash from spruce wood combustion. An increase of volatile substances at 340 degrees C gave rise to the suggestion that these compounds are re-formed by pyrolytic decomposition reactions from oligomeric, polymeric, and polyfunctional oxygenated species. This was corroborated by the investigation of the behavior of cellulose acetate, which exhibited a similar pattern in its SPI-TOFMS spectrum at 340 degrees C as the aerosol. More thorough investigations of urban aerosol and source material with respect to problems such as the mass closure of carbonaceous material, indications for source apportionment, and allotment of organic species on a molecular level to fractions of organic and elemental carbon seem feasible with this measurement method.     

Raman and near-infrared spectroscopy for quantification of fat composition in a complex food model system

Raman and near-infrared (NIR) spectroscopy have been evaluated for determining fatty acid composition and contents of main constituents in a complex food model system. A model system consisting of 70 different mixtures of protein, water, and oil blends was developed in order to create a rough chemical imitation of typical fish and meat samples, showing variation both in fatty acid composition and in contents of main constituents. The model samples as well as the pure oil mixtures were measured using Raman and NIR techniques. Partial least squares regression was utilized for prediction, and fatty acid features were expressed in terms of the iodine value and as contents of saturated, monounsaturated, and polyunsaturated fatty acids. Raman spectroscopy provided the best results for predicting iodine values of the model samples, giving validated estimation errors accounting for 2.8% of the total iodine value range. Both techniques provided good results for predicting the content of saturated, monounsaturated, and polyunsaturated fatty acids in the model samples, yielding validated estimation errors in the range of 2.4-6.1% of the total range of fatty acid content. Prediction results for determining fatty acid features of the pure oil mixtures were similar for the two techniques. NIR was clearly the best technique for modeling content of main constituents in the model samples.