Contribution of water-soluble extracts to the oxidative and inflammatory effects of atmospheric aerosols: A critical review.

Exposure to atmospheric particulate matter (PM) has been associated with heightened risks of lung cancer, cardiovascular and respiratory diseases. PM exposure also affects the immune system, leading to an increased susceptibility to infections, exacerbating pre-existent inflammatory and allergic lung diseases. Atmospheric PM can primarily impact human health through the generation of reactive oxygen species (ROS) that subsequently induce or exacerbate inflammation. These cytotoxic effects have been related with PM concentration, and its chemical constituents, including metals, solvent extractable organics (e.g., polycyclic aromatic hydrocarbons), and water-soluble ions. Although not receiving much attention, the fine aerosol water-soluble organic matter (WSOM) can account for a substantial portion of the overall fine PM mass and has been shown to present strong oxidative and immunomodulatory effects. Thus, the objective of this review is to provide a comprehensive analysis of the role of the water-soluble fraction of PM, with a specific focus on the contribution of the WSOM component to the cytotoxic properties of atmospheric PM. The chemical properties of the water-soluble PM fraction are briefly discussed, while emphasis is put on how PM size, composition, and temporal variations (e.g., seasonality) can impact the pro-oxidative activity, the modulation of inflammatory response, and the cytotoxicity of the water-soluble PM extracts.

Multidimensional chromatography in environmental analysis: Comprehensive two-dimensional liquid versus gas chromatography.

Analysis of complex environmental matrices poses an extreme challenge for analytical chemists due to the vast number of known and unknown compounds, with very diverse chemical and physical properties. The need for a holistic characterisation of this complexity has sparked the development of effective tools to unravel the chemical composition of such environmental samples. Multidimensional chromatographic methods, namely comprehensive two-dimensional (2D) gas and liquid chromatography (GC × GC and LC × LC, respectively), coupled to different detection systems have emerged as powerful tools with the capability to address this challenge. While GC × GC has steadily gained popularity in environmental analysis, LC × LC is surprisingly less attractive in this research field. This critical review article explores the potential reasons why LC × LC is not the dominant technique used in environmental analysis as compared to GC × GC, while simultaneously highlighting the quite unique role of LC × LC for the target and untargeted analysis of complex environmental matrices. The possible combinations of stationary phases, the important role of the interfacing valve as the heart of an LC × LC assembly, the existing optimization strategies for improving the separation power in the 2D chromatographic space, and the need for user-friendly mathematical tools for multidimensional data handling are also discussed. Finally, a set of practical measures are suggested to increase the use and secure the success of LC × LC in environmental analysis.

Source apportionment for indoor air pollution: Current challenges and future directions.

Source apportionment (SA) for indoor air pollution is challenging due to the multiplicity and high variability of indoor sources, the complex physical and chemical processes that act as primary sources, sinks and sources of precursors that lead to secondary formation, and the interconnection with the outdoor environment. While the major indoor sources have been recognized, there is still a need for understanding the contribution of indoor versus outdoor-generated pollutants penetrating indoors, and how SA is influenced by the complex processes that occur in indoor environments. This paper reviews our current understanding of SA, through reviewing information on the SA techniques used, the targeted pollutants that have been studied to date, and their source apportionment, along with limitations or knowledge gaps in this research field. The majority (78 %) of SA studies to date focused on PM chemical composition/size distribution, with fewer studies covering organic compounds such as ketones, carbonyls and aldehydes. Regarding the SA method used, the majority of studies have used Positive Matrix Factorization (31 %), Principal Component Analysis (26 %) and Chemical Mass Balance (7 %) receptor models. The indoor PM sources identified to date include building materials and furniture emissions, indoor combustion-related sources, cooking-related sources, resuspension, cleaning and consumer products emissions, secondary-generated pollutants indoors and other products and activity-related emissions. The outdoor environment contribution to the measured pollutant indoors varies considerably (<10 %- 90 %) among the studies. Future challenges for this research area include the need for optimization of indoor air quality monitoring and data selection as well as the incorporation of physical and chemical processes in indoor air into source apportionment methodology.

Assessing the relevance of allergenic pollen in indoor environments-current knowledge base and research needs.

Airborne pollen allergens-a relevant component of bioaerosols and, therefore, of airborne particulate matter-are considered an important metric in air quality assessments. Although the measurement of airborne pollen allergen concentrations in outdoor environments (namely, in urban areas) has been recognized as a key environmental health indicator, no such obligation exists for indoor environments (dwellings or occupational settings). However, people spend most of their daily time (80-90%) indoors, where the majority of their exposure to air pollution, including pollen allergens, occurs. Nonetheless, the relative importance of airborne pollen allergen exposure indoors differs from outdoors because of differences in pollen loads, sources, dispersion, and degree of penetration from the outdoor surroundings, as well as the differences in the allergenic pollen profiles. In this brief review, we mined the literature over the last 10 years to summarize what existing measurements reveal about the relevance of airborne allergenic pollen in indoor environments. The research priorities on this topic are presented, highlighting the challenges and the motivations for obtaining pollen data in built environments which are key to understand the extent and mechanisms of human exposure to airborne pollen allergens. Thus, we provide a comprehensive assessment of the relevance of airborne allergenic pollen in indoor environments, highlighting knowledge gaps and research needs related to their health effects.

Structural Features and Pro-Inflammatory Effects of Water-Soluble Organic Matter in Inhalable Fine Urban Air Particles.

The impact of inhalable fine particulate matter (PM2.5, aerodynamic diameter <2.5 µm) on public health is of great concern worldwide. Knowledge on their harmful effects are mainly due to studies carried out with whole air particles, with the contribution of their different fractions remaining largely unknown. Herein, a set of urban PM2.5 samples were collected during daytime and nighttime periods in autumn and spring, aiming to address the seasonal and day-night variability of water-soluble organic matter (WSOM) composition. In vitro analysis of the oxidative and pro-inflammatory potential of WSOM samples was carried out in both acute (24 h) and chronic (3 weeks) exposure setups using Raw264.7 macrophages as cell model. Findings revealed that the structural composition of WSOM samples differs between seasons and in a day-night cycle. Cell exposure resulted in an increase in the transcription of the cytoprotective Hmox1 and pro-inflammatory genes Il1b and Nos2, leading to a moderate pro-inflammatory status. These macrophages showed an impaired capacity to subsequently respond to a strong pro-inflammatory stimulus such as bacterial lipopolysaccharide, which may implicate a compromised capacity to manage harmful pathogens. Further investigation on aerosol WSOM could help to constrain the mechanisms of WSOM-induced respiratory diseases and contribute to PM2.5 regulations.

Effect of Soil Organic Matter, Soil pH, and Moisture Content on Solubility and Dissolution Rate of CuO NPs in Soil.

The objectives of this research were to quantify the impact of organic matter content, soil pH and moisture content on the dissolution rate and solubility of copper oxide nanoparticles (CuO NPs) in soil, and to develop an empirical model to predict the dissolution kinetics of CuO NPs in soil. CuO NPs were dosed into standard LUFA soils with various moisture content, pH and organic carbon content. Chemical extractions were applied to measure the CuO NP dissolution kinetics. Doubling the reactive organic carbon content in LUFA 2.1 soil increased the solubility of CuO NP 2.7-fold but did not change the dissolution rate constant. Increasing the soil pH from 5.9 to 6.8 in LUFA 2.2 soil decreased the dissolution rate constant from 0.56 mol1/3·kg1/3·s-1 to 0.17 mol1/3·kg1/3·s-1 without changing the solubility of CuO NP in soil. For six soils, the solubility of CuO NP correlated well with soil organic matter content ( R2 = 0.89) independent of soil pH. In contrast, the dissolution rate constant correlated with pH for pH < 6.3 ( R2 = 0.89), independent of soil organic matter content. These relationships predicted the solubility and dissolution rate constants of CuO NP in two test soils (pH 5.0 and pH 7.6). Moisture content showed negligible impact on the dissolution kinetics of CuO NPs. Our study suggests that soil pH and organic matter content affect the dissolution behavior of CuO NP in soil in a predictable manner.

Comparative study of atmospheric water-soluble organic aerosols composition in contrasting suburban environments in the Iberian Peninsula Coast.

This study investigates the structural composition and major sources of water-soluble organic matter (WSOM) from PM2.5 collected, in parallel, during summer and winter, in two contrasting suburban sites at Iberian Peninsula Coast: Aveiro (Portugal) and Coruña (Spain). PM10 samples were also collected at Coruña for comparison. Ambient concentrations of PM2.5, total nitrogen (TN), and WSOM were higher in Aveiro than in Coruña, with the highest levels found in winter at both locations. In Coruña, concentrations of PM10, TN, and WSOM were higher than those from PM2.5. Regardless of the season, stable isotopic δ13C and δ15N in PM2.5 suggested important contributions of anthropogenic fresh organic aerosols (OAs) at Aveiro. In Coruña, δ13C and δ15N of PM2.5 and PM10 suggests decreased anthropogenic input during summer. Although excitation-emission fluorescence profiles were similar for all WSOM samples, multi-dimensional nuclear magnetic resonance (NMR) spectroscopy confirmed differences in their structural composition, reflecting differences in aging processes and/or local sources between the two locations. In PM2.5 WSOM in Aveiro, the relative distribution of non-exchangeable proton functional groups was in the order: HC (40-43%) > HCC (31-39%) > HCO (12-15%) > Ar-H (5.0-13%). However, in PM2.5 and PM10 WSOM in Coruña, the relative contribution of HCO groups (24-30% and 23-29%, respectively) equals and/or surpasses that of HCC (25-26% and 25-29%, respectively), being also higher than those of Aveiro. In both locations, the highest aromatic contents were observed during winter due to biomass burning emissions. The structural composition of PM2.5 and PM10 WSOM in Coruña is dominated by oxygenated aliphatic compounds, reflecting the contribution of secondary OAs from biogenic, soil dust, and minor influence of anthropogenic emissions. In contrast, the composition of PM2.5 WSOM in Aveiro appears to be significantly impacted by fresh and secondary anthropogenic OAs. Marine and biomass burning OAs are important contributors, common to both sites.

Persistence of urban organic aerosols composition: Decoding their structural complexity and seasonal variability.

Organic Aerosols (OAs) are typically defined as highly complex matrices whose composition changes in time and space. Focusing on time vector, this work uses two-dimensional nuclear magnetic resonance (2D NMR) techniques to examine the structural features of water-soluble (WSOM) and alkaline-soluble organic matter (ASOM) sequentially extracted from fine atmospheric aerosols collected in an urban setting during cold and warm seasons. This study reveals molecular signatures not previously decoded in NMR-related studies of OAs as meaningful source markers. Although the ASOM is less hydrophilic and structurally diverse than its WSOM counterpart, both fractions feature a core with heteroatom-rich branched aliphatics from both primary (natural and anthropogenic) and secondary origin, aromatic secondary organics originated from anthropogenic aromatic precursors, as well as primary saccharides and amino sugar derivatives from biogenic emissions. These common structures represent those 2D NMR spectral signatures that are present in both seasons and can thus be seen as an "annual background" profile of the structural composition of OAs at the urban location. Lignin-derived structures, nitroaromatics, disaccharides, and anhydrosaccharides signatures were also identified in the WSOM samples only from periods identified as smoke impacted, which reflects the influence of biomass-burning sources. The NMR dataset on the H-C molecules backbone was also used to propose a semi-quantitative structural model of urban WSOM, which will aid efforts for more realistic studies relating the chemical properties of OAs with their atmospheric behavior.

Structural signatures of water-soluble organic aerosols in contrasting environments in South America and Western Europe.

This study describes and compares the key structural units present in water-soluble organic carbon (WSOC) fraction of atmospheric aerosols collected in different South American (Colombia - Medellín and Bogotá, Peru - Lima, Argentina - Buenos Aires, and Brazil - Rio de Janeiro, São Paulo, and Porto Velho, during moderate (MBB) and intense (IBB) biomass burning) and Western European (Portugal - Aveiro and Lisbon) locations. Proton nuclear magnetic resonance (1H NMR) spectroscopy was employed to assess the relative distribution of non-exchangeable proton functional groups in aerosol WSOC of diverse origin, for the first time to the authors' knowledge in South America. The relative contribution of the proton functional groups was in the order H-C > H-C-C= > H-C-O > Ar-H, except in Porto Velho during MBB, Medellín, Bogotá, and Buenos Aires, for which the relative contribution of H-C-O was higher than that of H-C-C=. The 1H NMR source attribution confirmed differences in aging processes or regional sources between the two geographic regions, allowing the differentiation between urban combustion-related aerosol and biological particles. The aerosol WSOC in Aveiro, Lisbon, and Rio de Janeiro during summer are more oxidized than those from the remaining locations, indicating the predominance of secondary organic aerosols. Fresh emissions, namely of smoke particles, becomes important during winter in Aveiro and São Paulo, and in Porto Velho during IBB. The biosphere is an important source altering the chemical composition of aerosol WSOC in South America locations. The source attribution in Medellín, Bogotá, Buenos Aires, and Lima confirmed the mixed contributions of biological material, secondary formation, as well as urban and biomass burning emissions. Overall, the information and knowledge acquired in this study provide important diagnostic tools for future studies aiming at understanding the water-soluble organic aerosol problem, their sources and impact at a wider geographic scale.

Tracing of aerosol sources in an urban environment using chemical, Sr isotope, and mineralogical characterization.

In the framework of two national research projects (ORGANOSOL and CN-linkAIR), fine particulate matter (PM2.5) was sampled for 17 months at an urban location in the Western European Coast. The PM2.5 samples were analyzed for organic carbon (OC), water-soluble organic carbon (WSOC), elemental carbon (EC), major water-soluble inorganic ions, mineralogical, and for the first time in this region, strontium isotope (87Sr/86Sr) composition. Organic matter dominates the identifiable urban PM2.5 mass, followed by secondary inorganic aerosols. The acquired data resulted also in a seasonal overview of the carbonaceous and inorganic aerosol composition, with an important contribution from primary biomass burning and secondary formation processes in colder and warmer periods, respectively. The fossil-related primary EC seems to be continually present throughout the sampling period. The 87Sr/86Sr ratios were measured on both the labile and residual PM2.5 fractions as well as on the bulk PM2.5 samples. Regardless of the air mass origin, the residual fractions are more radiogenic (representative of a natural crustal dust source) than the labile fractions, whose 87Sr/86Sr ratios are comparable to that of seawater. The 87Sr/86Sr ratios and the mineralogical composition data further suggest that sea salt and mineral dust are important primary natural sources of fine aerosols throughout the sampling period.

Two chemically distinct light-absorbing pools of urban organic aerosols: A comprehensive multidimensional analysis of trends.

The chemical and light-absorption dynamics of organic aerosols (OAs), a master variable in the atmosphere, have yet to be resolved. This study uses a comprehensive multidimensional analysis approach for exploiting simultaneously the compositional changes over a molecular size continuum and associated light-absorption (ultraviolet absorbance and fluorescence) properties of two chemically distinct pools of urban OAs chromophores. Up to 45% of aerosol organic carbon (OC) is soluble in water and consists of a complex mixture of fluorescent and UV-absorbing constituents, with diverse relative abundances, hydrophobic, and molecular weight (Mw) characteristics between warm and cold periods. In contrast, the refractory alkaline-soluble OC pool (up to 18%) is represented along a similar Mw and light-absorption continuum throughout the different seasons. Results suggest that these alkaline-soluble chromophores may actually originate from primary OAs sources in the urban site. This work shows that the comprehensive multidimensional analysis method is a powerful and complementary tool for the characterization of OAs fractions. The great diversity in the chemical composition and optical properties of OAs chromophores, including both water-soluble and alkaline-soluble OC, may be an important contribution to explain the contrasting photo-reactivity and atmospheric behavior of OAs.

Unraveling the structural features of organic aerosols by NMR spectroscopy: a review.

Our limited understanding of the effect of organic aerosols (OAs) on the climate and human health is largely because of the vast array of formation processes and sources that produce a multitude of molecular structures and physical properties. The need to unravel the enormous complexity and heterogeneity of OAs and thus understand their effects on the climate and human health has led to the development of different off-line methods based on the use of advanced analytical techniques. Within this context, nuclear magnetic resonance (NMR) spectroscopy has become essential for acquiring detailed structural characterization of the complex natural organic matter contained in atmospheric aerosols. In this article, we present a critical review on the application of NMR spectroscopy in OAs (primary and secondary) studies, focusing mainly on the water-soluble organic fraction, and how NMR has impacted our knowledge on atmospheric organic matter. A major emphasis is given on the wealth of chemical information that solid-state and multi-dimensional solution-state NMR can provide, including the sources, formation pathways, seasonal, and regional characterization of atmospheric OAs. Finally, major challenges are discussed and recommendations for future research directions are proposed.

A simple approach to reduce dimensionality from comprehensive two-dimensional liquid chromatography coupled with a multichannel detector.

Comprehensive two-dimensional liquid chromatographic (LC×LC) systems play an ever increasing role in separation and characterization of complex samples. When coupled with multichannel detectors, such as the diode array detector, these LC×LC systems become especially useful for non-target analysis and identification of patterns based on the information extracted from those complex samples. Nevertheless, due to the large amount of data generated by these systems, the extraction of useful information for the identification of patterns still is one of the major drawbacks for a wider application of this technique. As a preliminary step in data treatment, we have developed a simple and fast way to deal with this large amount of multi-dimensional data by identifying the three-dimensional (3D) regional maxima of each chromatographic peak generated in a LC×LC-DAD system: retention times at the peak maximum in the first- and second-dimensions and the wavelength of the maximum UV absorption. This dataset is then used to build a 3D fingerprinting of the given sample, which alongside the 3D fingerprinting of other samples, can be used to identify different patterns associated with the specific properties of every sample under study. The applicability of the developed methodology was further assessed by performing a non-target LC×LC-DAD analysis of four Portuguese red wine samples.

Determination of anionic surface active agents using silica coated magnetite nanoparticles modified with cationic surfactant aggregates.

The development of a novel methodology for extraction and preconcentration of the most commonly used anionic surface active agents (SAAs), linear alkylbenzene sulfonates (LAS), is presented herein. The present method, based on the use of silica-magnetite nanoparticles modified with cationic surfactant aggregates, was developed for determination of C10-C13 LAS homologues. The proposed methodology allowed quantitative recoveries of C10-C13 LAS homologues by using a reduced amount of magnetic nanoparticles. Limits of detection were in the range 0.8-1.9µgL(-1) for C10-C13 LAS homologues, while the repeatability, expressed as relative standard deviation (RSD), ranged from 2.0 to 3.9% (N=6). Finally, the proposed method was successfully applied to the analysis of a variety of natural water samples.

Humic acids as proxies for assessing different Mediterranean forest soils signatures using solid-state CPMAS 13C NMR spectroscopy.

Humic acids (HAs) of four representative forest soils profiles from Central Spain (two with different vegetation - pine and oak - but same parent material - granitie, and two with same vegetation - holm oak - but different parent material - granite and limestone) were investigated by solid-state cross polarization with magic angle spinning (13)C nuclear magnetic resonance (NMR) spectroscopy. The objectives included the investigation of the impact of different forest properties on HA composition, assessing how the structural characteristics of the HA vary with soil depth, and evaluating the role of HA as surrogates for mapping the different forest soils signatures using structural data derived from (13)C NMR spectroscopy. On average, alkyl C is the dominant C constituent (38-48% of the total NMR peak area) in all HA samples, followed by aromatic (12-22%) and O-alkyl C (12-19%), and finally carboxyl C (7.0-10%). The NMR data also indicated that HA composition is likely to be differently affected by the soil physico-chemical properties and type of forest vegetation. The structural characteristics of the HA from soil under oak did not differ broadly downward in the profile, whereas soil HA under pine forest exhibits a somewhat higher recalcitrant nature as a consequence of a higher degree of decomposition. The soil HA from holm oak forests differed from the other two forest soils, exhibiting a progressive decomposition of the alkyl C structures with increasing depth, while the carbohydrate-like indicator (O-alkyl C) is apparently being protected from mineralization in the horizons below the ground level. Overall, these differences in soil HA NMR signatures are an important diagnostic tool for understanding the role of different soil environmental factors on the structural composition of HA from Mediterranean forest soils.

A generalization of a chromatographic response function for application in non-target one- and two-dimensional chromatography of complex samples.

New flexible chromatographic response functions for one-dimensional (DCRF(f)) and two-dimensional chromatography (DCRF(f,2D)) are put forward and tested for estimating the quality index of separation in non-targeted chromatographic analysis of complex samples. These functions, based on already tested functions, have three clear-cut criteria: number of chromatographic peaks, degree of chromatographic separation, and time spent in the analysis. However, unlike the existing functions, they allow an accurate weighing and adjustment of these criteria according to the needs of the analyst. Special attention was given to the time spent in the analysis by introducing a time-saving criterion, which allowed its application in time dependent chromatographic systems. The performance and flexibility of the strategies based on the developed response functions are tested in different chromatographic scenarios and for several application possibilities using simulated chromatograms.

Trends in data processing of comprehensive two-dimensional chromatography: state of the art.

The operation of advanced chromatographic systems, namely comprehensive two-dimensional (2D) chromatography coupled to multidimensional detectors, allows achieving a great deal of data that need special care to be processed in order to characterize and quantify as much as possible the analytes under study. The aim of this review is to identify the main trends, research needs and gaps on the techniques for data processing of multidimensional data sets obtained from comprehensive 2D chromatography. The following topics have been identified as the most promising for new developments in the near future: data acquisition and handling, peak detection and quantification, measurement of overlapping of 2D peaks, and data analysis software for 2D chromatography. The rational supporting most of the data processing techniques is based on the generalization of one-dimensional (1D) chromatography although algorithms, such as the inverted watershed algorithm, use the 2D chromatographic data as such. However, for processing more complex N-way data there is a need for using more sophisticated techniques. Apart from using other concepts from 1D chromatography, which have not been tested for 2D chromatography, there is still room for new improvements and developments in algorithms and software for dealing with 2D comprehensive chromatographic data.

Resolving the chemical heterogeneity of natural organic matter: new insights from comprehensive two-dimensional liquid chromatography.

For the purpose of resolving the chemical heterogeneity of natural organic matter (NOM), comprehensive two-dimensional liquid chromatography (LC×LC) was employed for the first time to map the hydrophobicity versus molecular weight (MW) distribution of two well-known complex organic mixtures: Suwannee River Fulvic Acids (SR-FA) and Pony Lake Fulvic Acids (PL-FA). Two methods have been developed using either a conventional reversed-phase (RP) silica column or a mixed-mode hydrophilic interaction column operating under aqueous RP mode in the first dimension, and a size-exclusion column in the second dimension. The LC×LC fractions were screened on-line by UV at 254 nm, molecular fluorescence at excitation/emission wavelengths (λ(Exc)/λ(Em)) of 240/450 nm, and by evaporative light scattering. The MW distributions of these two NOM samples were further characterized by number (Mn) and weight (Mw) average MW, and by polydispersity (Mw/Mn). Findings suggest that the combination of two independent separation mechanisms is promising in extend the range of NOM separation. For the cases where NOM separation was accomplished, smaller Mw group fractions seem to be related to a more hydrophobic nature. Regardless of the detection method, the complete range of MW distribution provided by both comprehensive LC×LC methods was found to be lower than those reported in the literature.

A new chromatographic response function for assessing the separation quality in comprehensive two-dimensional liquid chromatography.

A new chromatographic response function (CRF(2D)) is proposed and tested for the estimation of the quality index of separation in comprehensive two-dimensional liquid chromatography (2D-LC) of complex organic mixtures. This objective function is based on the concept of peak purity for one-dimensional liquid chromatography, which has been redefined for 2D-LC. The new CRF(2D) also includes other separation quality criteria, namely the number of 2D peaks appearing in the chromatogram and the analysis time. To compute the peak purity for a given 2D peak, three important steps have been tackled in this study: (a) the development of an alternative algorithm for detecting 2D peaks automatically from real experimental 2D-LC data; (b) the application of a mathematical model to fit the obtained chromatographic data; and (c) the estimation of the volume of the overlapping region between two or more 2D peaks. The performance of the developed CRF(2D) was compared to that of an existing resolution measure, using simulated chromatograms. The capability of the new function to qualify the overall separation degree that it is attained under different chromatographic conditions was further assessed through a 2D-LC study of a mixture of four aromatic compounds.

Optimizing size-exclusion chromatographic conditions using a composite objective function and chemometric tools: application to natural organic matter profiling.

A novel strategy is suggested for developing a size-exclusion chromatography (SEC) method in order to assess the molecular size distribution of natural organic matter (NOM). This strategy is based on the use of a chromatographic response function (CRF) and a response surface based on a central composite experimental design. The CRF qualifies the resolution degree attained under different SEC conditions with respect to global resolution of the chromatogram, number of distinguishable peaks, and analysis time. Based on a SEC study with a mixture of known organic compounds, predictive polynomial and linear models were developed according to the practice of design of experiments. Additionally, a chromatographic response surface was used, describing the effect of pH, amount of organic solvent, and salt concentration of the mobile phase on the values of the CRF. This new strategy predicted a single linear effect of salt concentration over the quality of the chromatographic separation, and the influence of this experimental variable on the quality of size-exclusion separations of various NOM samples was further discussed. For the analysis of NOM, the optimum settings for the mobile phase composition were as follows: pH 8.5, 11% of acetonitrile, and 15 mM of sodium chloride. Under such analytical conditions, the measured molecular size distributions of water-soluble organic matter from atmospheric particles and aquatic fulvic acids compare reasonably well with the published molecular weight data. The newly developed strategy is a reliable alternative for accomplishing an accurate description of the size-exclusion profiles of NOM from different environments.