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Spectromicroscopy Mapping of Colloidal/Particulate Organic Matter

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Spectromicroscopy Mapping of Colloidal/Particulate Organic Matter Environ. Sci. Technol. 2007, 41, 7864-7869 investigate floc/aggregate structures in freshwater systems Spectromicroscopy Mapping of (8, 9) in particular, as well as to obtain morphological Colloidal/Particulate Organic Matter information on the colloidal fraction (1-1000 nm) in freshwater (10, 11) and groundwater systems (12, 13). in Lake Brienz, Switzerland Low nutrient or oligotrophic alpine lakes have attracted increased interest over the past few years as they are sensitive THORSTEN SCHA¨ FER,² indicators of climate change. Recent publications have shown VINCENT CHANUDET,³,§ that the properties of the catchment have a clear impact on FRANCIS CLARET,| , ⊥ AND the macroinvertebrate communities in these lakes, while the MONTSERRAT FILELLA*,³ influence of geographical patterns is minor (14). Large inputs Institut fuÈr Nukleare Entsorgung (INE), Forschungszentrum of suspended sediments and glacial silt/colloids may also Karlsruhe, P.O. Box 3640, D-76021 Karlsruhe, Germany, limit the spring phytoplankton peak in alpine lakes, which Department of Inorganic, Analytical, and Applied Chemistry, occurs in a number of other temperate lakes (15, 16). As part University of Geneva, Quai Ernest-Ansermet 30, CH-1211 of a large project focused on understanding the severe Geneva 4, Switzerland, Institut F.-A. Forel, University of productivity limitation conditions existing in a peri-alpine Geneva, Route de Suisse, 10, CH-1290 Versoix, Switzerland, lake (17)sLake Brienz, Switzerlands,submicrometer natural and CEA Saclay, CEA/DPC/SECR/LSRM, Gif sur Yvette, France organic matter (NOM), in particular, refractory organic matter (ROM) (18), and inorganic colloids (19, 20) have been studied for over a year in the lake and its two main tributaries, the Aare and LuÈtschine Rivers. Transmission electron microscopy (TEM), soft X-ray The focus of this study is on (i) the characterization of the scanning transmission X-ray microscopy (STXM), and NOM found in the lake and in its tributary rivers, both alone µ-FTIR spectromicroscopy were used to map colloidal/ and in association with inorganic colloids; (ii) inter- particulate material in an ultra-oligotrophic lake, Lake Brienz, comparison of the data obtained by the three different Switzerland, with a special focus on organic functionality. methods used, namely, TEM, STXM, and µ-FTIR microscopy; Within the statistical margin of error and the uncertainties and (iii) comparison of organic functional group spectral arising from the representativeness of the results, the research signatures found in the lake with those of the two rivers. reveals that organic material was associated with potassium- Materials and Methods rich inorganic colloids present in surface and deep water (depths of 1 and 100 m, respectively), which indicates Sampling Sites. Samples were collected from the LuÈtschine River (46°38′52′′N, 7°52′29′′E) and the Aare River (46°44′37′′N, a vertical transfer of aggregates by sedimentation. Pure ° ′ ′′ ° ′ ′′ organic colloids could only be detected in surface waters. 8 3 2 E) and from the middle of Lake Brienz (46 43 2 N, 7°56′59′′E) in July 2005. Lake water samples were collected In addition, correlation map analysis of synchrotron- at depths of 1 and 100 m using a membrane pump, with the based µ-FTIR and carbon K-edge STXM spectromicroscopic end of the tube directly attached to a multi-parameter ZuÈllig data using spectra from the Lu¨tschine and Aare Rivers HPT-D probe. River water samples were collected directly as target spectra revealed spectral similarities with organic into bottles at a depth of about 10 cm. Samples for dissolved components from both tributary rivers in deeper regions organic carbon (DOC) determination were collected in (100 m) of the lake. The results prove that STXM and µ-FTIR precombusted (3 h at 550 °C) glass bottles. Samples for ROM can characterize colloidal and particulate organic material and carbohydrate analysis were collected in clean polyeth- in low organic carbon systems. ylene bottles. Immediately after collection, all samples were acidified to pH 2 with Suprapur grade HCl and filtered through precombusted (3 h at 550 °C) 1.2 µm glass filters (Whatman Introduction GF/C filters) by vacuum filtration. All samples were stored in a cooler in double plastic bags and kept in a refrigerator The application of chemical microscopy to biological systems until measured. All standard and sample solutions were has benefited greatly from synchrotron light, particularly prepared with 18 MΩ cm Milli-Q water. through scanning transmission X-ray microscopy (STXM) Methods. DOC was determined by a high-temperature µ and synchrotron-based infrared microscopy ( -FTIR) (1). combustion method using a TOC 5000-A Shimadzu analyzer. STXM, which uses X-ray absorption spectroscopy (NEXAFS) Milli-Q water was used as the blank (0.00 mg C L-1 with a as the contrast medium, is a very powerful tool for analyzing SD less than 0.005 mg C L-1). fully hydrated samples such as colloids (2-4). Dynes and co-workers have recently applied this method to investigating Total dissolved carbohydrates were analyzed using a microbial biofilms and the metal/chlorhexadine speciation modified MBTH (3-methyl-2-benzothiazolinone hydrochlo- ride) method (21, 22). Calibration was performed with in them (5, 6). The combination with transmission electron -1 microscopy (TEM) gives additional structural information at glucose. Results are expressed as mg C L . Polysaccharide the highest resolution (7). TEM has been extensively used to morphology was assessed by TEM with specifically stained (0.1 mmol of Ruthenium Red) TEM grids (18). * Corresponding author phone: (+41 22) 379 6046; fax: (+4122) ROM was measured by following the adsorptive stripping 3796069; e-mail: [email protected]. voltammetry response of the complex formed by these ² Institut fuÈr Nukleare Entsorgung. compounds in the presence of trace amounts of Mo(VI) (23). ³ Department of Inorganic, Analytical, and Applied Chemistry, This method is particularly well-suited to the quantitative University of Geneva. determination of low concentrations of humic-type com- § Institut F.-A. Forel, University of Geneva. | CEA Saclay. pounds in fresh water. The same concentrations were ⊥ Present address: BRGM, Environment and Process Division, 3 obtained when using standard river fulvic (IHSS Suwannee Avenue Claude Guillemin, F-45060 Orleans Cedex 2, France. River fulvic acid standard (1S101F)) or humic (IHSS Suwannee 7864 9 ENVIRONMENTAL SCIENCE & TECHNOLOGY / VOL. 41, NO. 22, 2007 10.1021/es071323z CCC: $37.00 2007 American Chemical Society Published on Web 10/09/2007 Laboratories (BNL) in New York, undulator beamline X1A1, operated by the State University of New York at Stony Brook. The principle of this method is described in detail elsewhere (27, 28). Carbon K-edge spectra were recorded in a constant helium atmosphere using an undulator gap of 36.8 mm. The Fresnel zone plate utilized for carbon-edge measurements had a diameter of 160 µm and an outermost zone width (δ) of 45 nm. Energy calibration of the spherical grating monochromator was performed using the photon energy of the CO2 gas adsorption band at 290.74 eV (29). Infrared measurements were performed at the U10B beamline (NSLS, BNL) using a Spectra-Tech Continuum IR microscope coupled with a Nicolet Magna 860 FTIR. The microscope utilizes a dual remote masking aperture and FIGURE 1. Mineralogical composition of inorganic colloids in Lake × Brienz and its tributaries as determined by TEM-EDS-SAED analysis. matching 32 Schwatzchild objectives. Spectra were collected using a 10 × 10 aperture and by averaging 512 scans in the mid-IR range (800-4000 cm-1) per point in transmission mode at a resolution of 4 cm-1 using Atlµs software (Thermo Nicolet Instruments). STXM measurements yield information on optical density (OD). OD is defined as the product of sample thickness d, sample density F, and mass absorption coefficient µ(E), which is related to the quotient of the incident flux on the sample I0(E) and the flux detected behind the sample I(E) via )- ) F OD ln[I(E)/I0(E)] µ(E) d (1) Image stacks were obtained by taking images at different energies across the absorption-edge and aligning them using cross-correlation. After stack alignment, the XANES spectra were extracted (30). Image regions that contained no particles gave the I0(E) information. Given the time and cost constraints involved in STXM and µ-FTIR measurements, it is at present not possible to analyze enough samples to evaluate statistically the signifi- cance of the results obtained. However, since in this study STXM and µ-FTIR analysis were performed in the same sample grids used for TEM-EDS-SAED measurements and FIGURE 2. STXM carbon K-edge and potassium L2,3-edge spectra that TEM-EDS-SAED results fit well with the known com- 1 of cluster analysis from (A) Lutschine River and (B) Aare River position of the inorganic colloids in the lake, it can be assumed sample grids. Regions of extracted cluster spectra are indicated that STXM and µ-FTIR map reasonably well the main types by arrows in the images shown, which were taken at 280 eV. Number of organic matter present in this system. of colloids/particles was 80 for the Lu1tschine River clusters and 40 for the Aare River clusters. Results and Discussion River humic acid standard II (2S101H)) acids for calibration. TEM-EDS-SAED and NOM Analysis. The colloid mineral- Results are expressed as milligrams of C per liter. ogical compositions identified by TEM-EDS-SAED analysis The chemical and mineralogical composition of inorganic are shown in Figure 1. The inorganic minerals present in colloids was assessed by TEM coupled with energy dispersive Lake Brienz and its tributaries are dominated by clay-like, spectroscopy (EDS) and selected area electron diffraction flattened colloidal particles (Figure S1, Supporting Informa- (SAED). Specimen grids were prepared on-site by using a tion): illite, chlorite, biotite, and Ti-rich biotite, but also by non-perturbing procedure based on the centrifugation of albite, orthose, and quartz.
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