Lilian R. F. Carvalho

Mutagenicity and DNA adduct formation of PAH, nitro-PAH, and oxy-PAH fractions of atmospheric particulate matter from São Paulo, Brazil.

Urban particulate matter (UPM) contributes to lung cancer incidence. Here, we have studied the mutagenic activity and DNA adduct-forming ability of fractionated UPM extractable organic matter (EOM). UPM was collected with a high-volume sampler in June 2004 at two sites, one at street level adjacent to a roadway and the other inside a park within the urban area of the city of São Paulo, Brazil. UPM was extracted using dichloromethane, and the resulting EOM was separated by HPLC to obtain PAH, nitro-PAH, and oxy-PAH fractions which were tested for mutagenicity with the Salmonella strains TA98 and YG1041 with and without S9 metabolic activation. The PAH fraction from both sites showed negligible mutagenic activity in both strains. The highest mutagenic activity was found for the nitro-PAH fraction using YG1041 without metabolic activation; however, results were comparable for both sites. The nitro-PAH and oxy-PAH fractions were incubated with calf thymus DNA under reductive conditions appropriate for the activation of nitro aromatic compounds, then DNA adduct patterns and levels were determined with thin-layer chromatography (TLC) 32P-postlabeling method using two enrichment procedures-nuclease P1 digestion and butanol extraction. Reductively activated fractions from both sites produced diagonal radioactive zones (DRZ) of putative aromatic DNA adducts on thin layer plates with both enrichment procedures. No such DRZ were observed in control experiments using fractions from unexposed filters or from incubations without activating system. Total adduct levels produced by the nitro-PAH fractions were similar for both sites ranging from 30 to 45 adducts per 10(8) normal nucleotides. In contrast, the DNA binding of reductively activated oxy-PAH fractions was three times higher and the adduct pattern consisted of multiple discrete spots along the diagonal line on the thin layer plates. However, DNA adduct levels were not significantly different between the sampling sites. Both samples presented the same levels of mutagenic activity. The response in the Salmonella assay was typical of nitroaromatics. Although, more mutagenic activity was related to the nitro-PAH fraction in the Salmonella assay, the oxy-PAH fractions showed the highest DNA adduct levels. More studies are needed to elucidate the nature of the genotoxicants occurring in São Paulo atmospheric samples.

Genotoxicity of fractionated organic material in airborne particles from São Paulo, Brazil.

Particulate matter less than 10 microns aerodynamic diameter (PM10) is associated with adverse health effects including increased respiratory problems and mortality. PM10 is also associated with increases in cancer in some urban areas. Identification of toxic compounds in PM10 is a step toward estimating exposure to these compounds and evaluating their public health risk. However, the toxic compounds on PM10 are part of a highly complex mixture of compounds that makes chemical characterization difficult. Before this study, there has been little investigation of genotoxic compounds in particulate matter from Latin American cities. Here, both bioassay (mutagenicity) and chemical analyses were conducted with organic solvent extracts of PM10 collected from São Paulo, a major Brazilian city. Sequential extraction in dichloromethane (DCM) followed by acetone (ACE) yielded 20.3% and 10.2% of the total mass, respectively. Non-polar and moderately polar organic material solubilized in DCM. ACE extracted more polar organic species and some inorganic ions. Both extracts were fractionated separately using cyanopropyl-bonded silica chromatography with organic solvents of increasing polarity. The mass distribution among the fractions was measured. The mutagenic activity of the fractions was assayed using the microsuspension procedure with the Salmonella typhimurium tester strain TA98, with and without addition of metabolic enzymes (S9). The DCM extract had about four times higher mutagenic activity than the ACE extract. In general, addition of S9 resulted in an increase in mutagenicity of DCM fractions, but a decrease for the ACE extract. Most of the activity was concentrated in fractions in the mid-range of polarity within both the DCM and ACE extracts. The fractions were analyzed by gas chromatography with mass selective detection (GC/MS) without derivatization. The most mutagenic fractions in the DCM extract contained ketones, aldehydes, and quinolines. The most mutagenic ACE fraction had ketones, carboxylic acids, and aldehydes.

An artifact in air carbonyls sampling using C18 DNPH-coated cartridge

A sampling artifact is observed when collecting carbonyl compounds in ambient air using 2,4-dinitrophenylhydrazine (DNPH)-coated C18 cartridges. While lower carbonyls (C1–C4) are not significantly affected, ca. 80% of C5–C8 carbonyl concentrations are due to artifact formation. Furthermore, several high molecular weight carbonyls, containing up to 18 carbons, were artificially produced during sampling probably due to reactions between ambient ozone and C18 sorbent matrix. The use of ozone trap is recommended for carbonyls sampling with DNPH-coated solid sorbents. The most suitable sampling system is a DNPH-coated silica gel cartridge connected to an upstream crystalline KI trap.

Monitoring of the ultrasonic irradiation effect on the extraction of airborne particulate matter by ion chromatography

The action of ultrasonic waves in liquids can induce or accelerate chemical reactions of the liquids and/or substances dissolved. A study on ultrasonic irradiation effects in aqueous and acetone extracts from airborne particulate matter, was developed. Several systems such as solvents, filters, standard reference material of airborne particles and real samples from atmospheric particulate matter were evaluated. The generation of anionic species during the sonication process was confirmed by ion chromatography. The change of quantitative and qualitative data has shown that ultrasonic extraction leads to false interpretation of analytical results.

A preliminary characterization of the mutagenicity of atmospheric particulate matter collected during sugar cane harvesting using the Salmonella/microsome microsuspension assay.

During sugar cane harvesting season, which occurs from May to November of each year, the crops are burnt, cut, and transported to the mills. There are reports showing that mutagenic activity and PAH content increase during harvesting season in some areas of São Paulo State in comparison with nonharvesting periods. The objective of this work was to preliminarily characterize the mutagenic activity of the total organic extracts as well as corresponding organic fractions of airborne particulate matter (PM) collected twice from two cities, Araraquara (ARQ) and Piracicaba (PRB), during sugar cane harvesting season using the Salmonella/microsome microssuspension assay. One sample collected in São Paulo metropolitan area was also included. The mutagenicity of the total extracts ranged from 55 to 320 revertants per cubic meter without the addition of S9 and from not detected to 57 revertants per cubic meter in the presence of S9 in areas with sugar cane plantations. Of the three fractions analyzed, the most polar ones (nitro and oxy) were the most potent. A comparison of the response of TA98 with YG1041 and the increased potencies without S9 indicated that nitro compounds are causing the observed effect. More studies are necessary to verify the sources of the mutagenic activity such as burning of vegetal biomass and combustion of heavy duty vehicles used to transport the sugar cane to the mills. The Salmonella/microsome assay can be an important tool to monitor the atmosphere for mutagenicity during sugar cane harvesting season. Environ. Mol. Mutagen. 2008.

Systematic approach to the separation of mono- and hydroxycarboxylic acids in environmental samples by ion chromatography and capillary electrophoresis

Abstract Strategies for method development and optimization of conditions in ion chromatography (IC) and capillary electrophoresis (CE) are proposed. The approaches are based on well established theoretical models for analyte elution and rely on the inspection of effective capacity factor versus specific gradient ramp slope curves for IC and effective mobility versus pH curves for CE. Commonly used IC eluent systems, such as sodium hydroxide and sodium borate, and the CE electrolyte 3,5-dinitrobenzoic acid were investigated. A standard aqueous solution containing formate, acetate, α-hydroxyacetate (glycolate), propionate, α-hydroxypropionate (lactate), butyrate and β-hydroxybutyrate anions was used to test the analytical conditions indicated by the models. Among the IC eluent systems, borate presented a better separation performance than hydroxide. However, a complete resolution of all components in the standard mixture was not accomplished experimentally at the chosen gradient ramp slope of 0.05 mM/ml. The analyte pairs acetate/lactate and glycolate/butyrate, whose effective capacity factors differ by approximately 0.2 units, co-eluted at 0.17 and 0.24 resolution, respectively, and the retention time of the last eluting analyte was relatively long (20 min). Nevertheless, the IC method provided the best overall limit of detection (LOD; 0.016–0.082 mg/l). Under the optimized CE conditions, all seven components in the standard mixture were resolved satisfactorily in less than 7 min. The analyte pair β-hydroxybutyrate/butyrate presented the worst resolution, 0.45, and a difference in effective mobility of 1.6%. The CE methodology provided the best column efficiency, roughly a ten-fold improvement in terms of number of plates per meter over the IC method, but the limit of detection was comparatively poorer (0.050–0.36 mg/l). Both proposed IC and CE methodologies can be applied to the analysis of mono- and hydroxycarboxylic acids in samples of environmental interest, providing complementary information. The choice of the most appropriate method is a compromise between chemical composition of the sample and concentration level of the analytes under investigation. For instance, the classical co-elution of acetate/lactate that occurs in IC columns, can be solved by CE. But the CE method may lack sensitivity for these analytes, which compromises the analysis of certain real samples. The LOD for acetate and lactate as determined by IC is 0.036 and 0.082 mg/l, respectively, while for CE they increase to 0.11 and 0.20 mg/l. Under optimum conditions, the separation and identification of mono- and hydroxycarboxylic acids in an atmospheric particulate matter sample is illustrated by both techniques.

Environmental formaldehyde analysis by active diffusive sampling with a bundle of polypropylene porous capillaries followed by capillary zone electrophoretic separation and contactless conductivity detection

Compared to other volatile carbonylic compounds present in outdoor air, formaldehyde (CH(2)O) is the most toxic, deserving more attention in terms of indoor and outdoor air quality legislation and control. The analytical determination of CH(2)O in air still presents challenges due to the low-level concentration (in the sub-ppb range) and its variation with sampling site and time. Of the many available analytical methods for carbonylic compounds, the most widespread one is the time consuming collection in cartridges impregnated with 2,4-dinitrophenylhydrazine followed by the analysis of the formed hydrazones by HPLC. The present work proposes the use of polypropylene hollow porous capillary fibers to achieve efficient CH(2)O collection. The Oxyphan fiber (designed for blood oxygenation) was chosen for this purpose because it presents good mechanical resistance, high density of very fine pores and high ratio of collection area to volume of the acceptor fluid in the tube, all favorable for the development of air sampling apparatus. The collector device consists of a Teflon pipe inside of which a bundle of polypropylene microporous capillary membranes was introduced. While the acceptor passes at a low flow rate through the capillaries, the sampled air circulates around the fibers, impelled by a low flow membrane pump (of the type used for aquariums ventilation). The coupling of this sampling technique with the selective and quantitative determination of CH(2)O, in the form of hydroxymethanesulfonate (HMS) after derivatization with HSO(3)(-), by capillary electrophoresis with capacitively coupled contactless conductivity detection (CE-C(4)D) enabled the development of a complete analytical protocol for the CH(2)O evaluation in air.

ALQUILBENZENO SULFONATO LINEAR: UMA ABORDAGEM AMBIENTAL E ANALÍTICA

The environmental impact of detergents and other consumer products is behind the continued interest in the chemistry of the surfactants used. Of these, linear alkylbenzene sulfonates (LASs) are most widely employed in detergent formulations. The precursors to LASs are linear alkylbenzenes (LABs). There is also interest in the chemistry of these hydrocarbons, because they are usually present in commercial LASs (due to incomplete sulfonation), or form as one of their degradation products. Additionally, they may be employed as molecular tracers of domestic waste in the aquatic environment. The following aspects are covered in the present review: The chemistry of surfactants, in particular LAS; environmental impact of the production of LAS; environmental and toxicological effects of LAS; mechanisms of removal of LAS in the environment, and methods for monitoring LAS and LAB, the latter in domestic wastes. Classical and novel analytical methods employed for the determination of LAS and LAB are discussed in detail, and a brief comment on detergents in Brazil is given.

Volatile organic compounds inside urban tunnels of São Paulo City, Brazil

Volatile organic compounds, such as oxygenated hydrocarbons, aliphatic and aromatic hydrocarbons were measured in ambient air from two distinct urban tunnels in Sao Paulo City, Brazil, in special, at rush hours. In these tunnels, traffic jams with significant numbers of vehicles with inadequate emission controls are frequent. Sao Paulo, a highly polluted city, has an unconventional mix of vehicle types in that a variety of gasoline blends, including oxygenated ones are used. Inside and outside tunnel measurements were performed to evaluate pollutant levels originated from direct vehicular emissions and formed by atmospheric photochemistry. Formaldehyde/acetaldehyde ratios of tunnel (A) circulating only light-duty vehicles fueled mainly with gasohol, a mixture containing 78-80% (v/v) gasoline and 20-25% ethanol, were < 1; ratios of tunnel (B) circulating light-duty vehicles fueled mainly with gasohol and heavy diesel vehicles, were approximately equal to 1. Formic acid/acetic acid ratios of both tunnels were much lower than 1. Benzaldehyde was suggested as a possible tracer of light-duty gasohol vehicular emissions. n-alkanes were the most abundant compound class (similar levels in both tunnels) followed by carbonyl compounds (higher levels in B), and aromatics compounds (higher levels in A). Organic acids, the minor species, were found at similar levels in both tunnels. Results of this survey show that toxic pollutants are present at significant levels in both tunnels. Formaldehyde and benzene, human carcinogenic agents, presented maximum mixing ratios of 39.2 and 24.8 ppbv, respectively, inside tunnel A; acetaldehyde, a probable human carcinogenic, reached a maximum of 34.5 ppbv inside tunnel B. Since vehicle occupants that spend a significant time in urban tunnels are commonly exposed to toxic pollutant emissions, future extensive studies should be done in order to estimate exposure levels of toxic ambient air pollutants for these people.

Avaliação dos métodos analíticos para a determinação de metabólitos do benzeno como potenciais biomarcadores de exposição humana ao benzeno no ar

Increasing attention is being paid to the use of biomarkers for determining the exposure of humans to air toxics. Biomarkers include the nonreacted toxic substance, their metabolites, or the reaction products of these toxics with naturally substances in the body. Significant progress has been made in the measurement of biomarkers during the past several years. Much of this progress has been because of the development of advanced analytical techniques for identification and quantification of the chemical species in complex matrix, such as biological fluids. The assessment of the potential cancer risk associated with exposure to benzene at occupational and non-occupational ambient is necessary because of the toxicological implications of this air pollutant. Thus, in this review, the analytical methodologies used to determine the benzene metabolites, in special, urinary muconic acid and S-phenylmercapturic acid, are described and several problems affecting the precision of these procedures are discussed. Finally, in view of the difficulty pointed out for selecting the more adequate biomarker, further studies to evaluate the human exposure levels to benzene should be done.