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The present work focused on the development of an analytical method suitable to study the presence and the release of organophosphate esters (OPEs) and perfluoroalkylated substances (PFASs) from food contact materials (FCMs), from baking paper and aluminium foil. Although these classes of compounds are attracting increasing attention due to their toxicity the knowledge is still insufficient. The extent of their migration from FCMs to food was estimated using different liquid simulants. Ethanol 95 % was used to simulate the contact with fatty food, whilst acetic acid 3 % to mime contact with acidic aqueous-based food and the contact mode involved the use of the ultrasound-assisted technique. Preliminary results showed the higher migration for baking paper samples in contact with aqueous simulant with contaminations in the ranges 78.30-413.21 ng/dm2 and 1.43-13.87 ng/dm2 for OPEs and PFASs respectively. These findings highlighting the need to monitor particularly OPEs in FCMs.

Background: The determination of mancozeb, a fungicide extensively used in agriculture, is a challenge, due to the nature of the compound, a manganese and zinc complex of ethylenebis dithiocarbamate and because of the general instability of the dithiocarbamates. Methods: Mancozeb was analyzed in a GC-EI-MS system after derivatization by CE-UV with detection at 280 nm and in LC-ESI-MS-MS in MRM mode. Results: A comparative study of the performance of three different techniques for the detection of mancozeb was explored, highlighting the advantages and drawbacks of them. The limits of detection and quantification of the techniques were determined; the repeatability was assessed, showing values of relative standard deviation. Gas chromatography, although very sensitive, was not reproducible enough due to fast degradation of the derivatization product, whereas capillary electrophoresis-UV showed problems in run-to-run reproducibility which had the worst limit of detection. LC coupled with tandem mass spectrometry was the most reliable and precise technique and was able to determine the main degradation product of Mancozeb, at the same time. The proposed LC procedure was verified by applying it to a commercial formulation, a fungicide of known concentration, and to Italian white grapes treated with the formulation sprayed during cultivation. Conclusion: Thanks to the simplified sample handling, the proposed method resulted to be simple, fast, green, economic, and suitable for residue analysis in grapes and other fruits. Finally, the method was compared with other similar investigations.

The lack of information on biological risks in workplaces arises from the difficulty to measure bioaerosol. This study aimed to develop and improve the bioaerosol monitoring technique that uses proper biomarkers as a tool. Muramic and dipicolinic acids, and ergosterol were used as tracers for bacteria cells, bacterial spores, and fungal spores, respectively. Furthermore, 12- and 13-methyltetradecanoic acids (iso- and anteiso- C15:0) were used to study the presence of airborne bacteria and 3-hydroxy fatty acids were used to determine the concentration of peptidoglycan. Airborne particulate matter was sampled in a municipal indoor waste composting facility by multistage impactor samplers, during three main stages of composting process. The microorganism content, in airborne particles with aerodynamic diameter minor then 1 μm and between 1 and 10 μm, was determined starting from the aforementioned biomarker concentrations. For iso- and anteiso- C15:0, a conversion factor to transform its concentration into bacterial content was tentatively proposed. The results show that the chemical method covers some gaps in the information about bioaerosol presence in polluted atmospheres. Differences up to two orders of magnitude are observed, by comparing the results obtained by biomarkers and by cultivation-dependent methods. The microbial content, expressed as a percentage by mass on respect the PM, ranged from 4 to 28% with higher percentages during shredding and mixing stages and lower values during biocell opening operations. Bacterial spores, bacterial cells, and fungal spores detected were high in number, compared with the findings in similar studies elsewhere.

Mycotoxins’ exposure by inhalation and/or dermal contact can occur in different branches of industry especially where heavily dusty settings are present and the handling of dusty commodities is performed. This study aims to explore the possible contribution of the occupational exposure to aflatoxins by analysing urine samples for the presence of aflatoxins B1 and M1 and aflatoxin B1-N7-guanine adduct. The study was conducted in 2017 on two groups of volunteers, the workers group, composed by personnel employed in an Italian feed plant (n = 32), and a control group (n = 29), composed by the administrative employees of the same feed plant; a total of 120 urine samples were collected and analysed. A screening method and a quantitative method with high-resolution mass spectrometry determination were developed and fully validated. Limits of detections were 0.8 and 1.5 pg/mLurine for aflatoxin B1 and M1, respectively. No quantitative determination was possible for the adduct aflatoxin B1-N7-guanine. Aflatoxin B1 and its adduct were not detected in the analysed samples, and aflatoxin M1, instead, was found in 14 samples (12%) within the range 1.9–10.5 pg/mLurine. Only one sample showed a value above the limit of quantification (10.5 pg/mLurine). The absence of a statistical difference between the mean values for workers and the control group which were compared suggests that in this specific setting, no professional exposure occurs. Furthermore, considering the very low level of aflatoxin M1 in the collected urine samples, the contribution from the diet to the overall exposure is to be considered negligible.

As epidemiological studies have shown a significant association between exposure to particles and mortality and morbidity, depending on their different deposition efficiency in the pulmonary region, size-segregated particulate matter (PM) samples were collected in workplaces where high PM concentrations were expected and compared with the results from urban and background atmospheres. We investigated the following workplaces: an analytical chemistry laboratory, a wastewater treatment plant, a livestock farming activity, a municipal solid waste composting plant, and a waste electrical and electronic equipment (WEEE) Treatment Facility. The highest PM concentrations were found in the last three sites. Ultrafine PM (PM0.1: aerodynamic diameter less than 0.1 μm) mass concentrations also proved to be especially high in the livestock farming activity, in the municipal solid waste composting plant, and in the WEEE treatment facility, ranging between 4.7 and 19.8 μg m−3 as mean values, suggesting that this particulate fraction should be monitored and controlled. On the other hand, in both natural and urban environments, ultrafine particles occurred at concentrations 0.1–0.3 times the workplace values. It would have been interesting to determine the increase in cancer risk due to ultrafine particles, but cohort studies on PM0.1 are lacking. Therefore, effects on the workers were investigated starting from the lifetime average daily PM2.5 dose. Even with this limitation, the results show that people working in municipal solid waste composting plant and in a WEEE Treatment Facility are exposed to a not negligible additional lifetime risk on respect citizens.

The coupling of Thermogravimetry (TG) in conjuction with chemometrics was investigated for the first time to evaluate the capabilities of this novel test to provide the screening of thalassemia in blood samples stored at 4 °C until 15 days. Healthy donors were considered as reference subjects and a typical thermal behaviour as a function of aging was estimated and compared to thermal behaviour of thalassemia subjects. Diagnosis of thalassemia was made at the Day-Hospital Thalassemia of S. Eugenio Hospital through a comprehensive assessment of clinical presentation and hematological and molecular analysis. Despite blood changes with aging, the application of the TGA/Chemometrics test reveal that healthy and thalassemic population may be significantly differentiated after 15 days from blood collection with a 100% of correct classification rate. This new method applied to aged samples was able to discriminate thalassemia in transfused patients that is generally not possible by the common first level protocol used for thalassemia screening, and in δβ-thalassemias, and β-thalassemia combined with Hb Lepore, usually requiring the molecular analysis for diagnosis. This study, for the first time, describes a screening method for thalassemia able to detect thalassemia on whole blood samples stored for 15 days. In conclusion TGA/Chemometrics screening test for thalassemia is not influenced by the aging of blood samples and this approach could open the way to prevention programs for thalassemia also in the developing countries where expertise and facilities for their control are extremely limited or in situation involving delayed analyses.

In the last years, many studies have focused on risk assessment of exposure of workers to airborne particulate matter (PM). Several studies indicate a strong correlation between PM and adverse health outcomes, as a function of particle size. In the last years, the study of atmospheric particulate matter has focused more on particles less than 10 μm or 2.5 μm in diameter, however, recent studies identify in particles less than 0.1 μm the main responsibility for negative cardiovascular effects. The present paper deals with the determination of 66 organic compounds belonging to six different classes of persistent organic pollutants (POPs) in the ultrafine, fine and coarse fractions of PM (PM &lt, 0.1 µm, 0.1 &lt, PM &lt, 2.5 µm and 2.5 &lt, 10 µm) collected in three outdoor workplaces and in an urban outdoor area. Data obtained were analyzed with principal component analysis (PCA), in order to underline possible correlation between sites and classes of pollutants and characteristic emission sources. Emission source studies are, in fact, a valuable tool for both identifying the type of emission source and estimating the strength of each contamination source, as useful indicator of environment healthiness. Moreover, both carcinogenic and non-carcinogenic risks were determined in order to estimate human health risk associated to study sites. Risk analysis was carried out evaluating the contribution of pollutant distribution in PM size fractions for all the sites. The results highlighted significant differences between the sites and specific sources of pollutants related to work activities were identified. In all the sites and for all the size fractions of PM both carcinogenic and non-carcinogenic risk values were below acceptable and safe levels of risks recommended by the regulatory agencies.

Several studies in literature have highlighted associations between airborne particulate matter and several adverse health outcomes, as a function of particle size. Traditionally, PM studies have focused on particles less than 10 μm in diameter (PM10) or particles less than 2.5 μm in diameter (PM2.5), with each fraction characterized by a distinct source, and different composition and health effects. Since, diffusion in the alveolar region of smaller particles with an aerodynamic diameter less than 0.1 μm (i.e. 100 nm and therefore also defined as nanoparticles) becomes an effective mechanism and the probability of deposition increases. There is a great debate whether ultrafine fraction (PM0.1) is mainly responsible for negative cardiovascular effects caused by its high oxidative and mutagenic potential. In the present study, size-fractionated airborne particulate matter was collected from outdoor urban and working environments and analyzed for 105 organic contaminants of different classes: polycyclic aromatic hydrocarbons (PAH) and their derivatives (nitro-PAH and oxy-PAH); polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs) and Novel Brominated flame-retardants (N-BFRs) selected for their toxicity and persistence in the environment. The selected organic contaminants were subsequently divided into three different macro-groups based on chemical and physical properties (PAH/N-PAH/O-PAH; PCB and PBDE/N-BFR) and subjected to statistical analysis. The monitoring campaigns were carried out in four sites: an urban atmosphere (RM), a wastewater treatment plant (WWTP) where aerosol is generated during the different phases of the processes, an intensive livestock farming activity, characterized by sheds serving as a shelter for cows (COW) and an area where feed is stored (FEED). In each monitoring campaign PM was collected with a multistage low-pressure impactor able to sample 14 size intervals of PM on as many filters, subsequently joined to form three dimensional fractions (coarse, fine and ultrafine). The results obtained from the organic contaminant analyses and from the Principal Component Analysis (PCA) showed a correlation between sites and classes of pollutants, allowing the identification of characteristic emission sources of each monitored site. Emission source studies are in fact a valuable tool for both identifying the type of emission source characteristic of a specific place and estimating the strength of each contamination source in the same place of interest.

This paper provides a method for the quantification of sterols in different types of calf feedstuffs based on soy, sunflower, hay, calf feed and a mixture of all of them. The free fraction and the total sterolic fraction, after saponification and acidic hydrolysis of the samples, are extracted by solvent and the sterols are identified/quantified by reversed phase HPLC coupled to tandem mass spectrometry by atmospheric pressure chemical ionization. After the recovery evaluation, the method is validated in terms of linearity (coefficient of determination R2), repeatability (coefficient of variation RSD), limit of detection and quantification. In most of the cases, the most representative phytosterol is β-sitosterol, followed by campesterol or stigmasterol and by other minor sterols such as fucosterol, and Δ-5-avenasterol. In addition, also cholesterol and ergosterol, if present, are evaluated in all the samples. As far as we know, very little information is available on the investigated feeds, which are commonly used on farms. The results of this survey were compared to other studies, if present in literature, showing good agreement. The proposed method resulted to be simple, fast and suitable for application to other sterols, feedstuffs and derived foods. The knowledge of the sterolic content and composition is getting more and more important, both in terms of comprehension of the vegetal biochemistry and as basis for sterolomic studies.

The biomass burning marker levoglucosan is produced via combustion of woody cellulose at temperature over 300 °C, and is hence widely used to trace fires or discriminate urban air pollution sources. In this context, it is very useful the availability of a standard reference material (SRM) with a certified value of levoglucosan, to check the quality, reliability and accuracy of the measurements. To this aim, in this work a simple and rapid method was developed for the detection and quantification of levoglucosan in an urban dust Standard Reference Material 1649a, available from the National Institute of Standards and Technology and the results were compared to those obtained in our previous investigations. 5 mg of SRM particulate material were analyzed, after an ultrasonic extraction with deionized water, using a column, with stationary phase based on hydrophilic interaction, assembled on a liquid chromatography system combined with triple tandem quadrupole mass spectrometry (HPLC-MS/MS) with electrospray ionization. Unlike existing methods, it does not require neither derivatization or extensive clean-up of the sample. For the method validation, the matrix effect, limit of detection (LOD), limit of quantification (LOQ), linearity, accuracy, recovery and precision were evaluated. The concentration determined in the SRM was in good agreement with that determined by gas chromatography-mass spectrometry in our previous study. The precision, tested on SRM, over five replicates and five aliquots were within 15%, whereas accuracy ranged between 85 and 115%. To demonstrate the method is fit for the purpose, particulate matter samples collected in an urban area of the city of Rome Italy were analyzed. The amount of levoglucosan was consistent with the urban emissions typical of the considered season.

In this work, the application of a novel strategy to extend the shelf life of fresh cut fruits isproposed. In particular, edible film coatings from natural resources were used with the aim ofextending the shelf life of fresh cut apples. Chitosan and carboxymethylcellulose were consideredand a promising formulation was developed. The evaluation of the shelf life extension of fruits wasperformed in a multiparametric way over a period of 10 days. The percentage weight losses werecalculated for treated and untreated fruits in order to evaluate the performances of the coatingmaterialto avoid contact with environment. In the meanwhile, gas chromatography coupled to massspectrometry (GC-MS) was considered to determine the volatile compounds producing acharacteristic flavor of apples. Results demonstrated a significant improvement in the physiologicalloss in weight of treated fruits with respect to controls, while the study of the volatile componentindicated the ability of treated fruits to retain its freshness during storage.

A quick and reproducible screening analytical method for the simultaneous determination of algal toxins, belonging to different chemical classes, was developed to provide a toxin profile in seawater, useful to assess potential risks to environment and human health. The target compounds were: gonyautoxin-1,4, gonyautoxin-2,3, decarbamoylgonyautoxin-2,3, N‐sulfocarbamoyl-gonyautoxin-1,2, neosaxitoxin, decarbamoyl-neosaxitoxin, saxitoxin, decarbamoyl-saxitoxin, domoic acid, 13-desmethyl spirolide C (SPX1), palytoxin and okadaic acid. Extraction and clean-up were carried out with a combination of Bond Elut LRC-C18 and Carbograph4 cartridges connected in series. Analyte separation was performed in gradient elution mode in 12 min with a Gemini C18 column. Compound detection was carried out in multiple reaction monitoring and in positive ionization mode for paralytic shellfish poisoning toxins, domoic acid, SPX1 and palytoxin, and in negative ionization mode for okadaic acid. The toxins were quantified with matrix-matched calibration curves constructed by spiked seawater samples (concentration levels 0.02–2 μg L−1 depending on the compound). The method was reproducible with intra-day and inter-day relative standard deviations ranging from 4 to 9% and from 8 to 16%, respectively. Good recoveries (84–105%) and good accuracy (2–20%) were obtained by spiking experiments. Limits of detection were calculated for each toxin and varied from 0.011 to 0.12 μg L−1 depending on the compound. The developed method was applied to cultured Ostreopsis cf. ovata samples. The proposed procedure may be considered a valuable alternative to existing methods for monitoring toxic microalgae, since it offers a rapid screening of target toxins, reduced organic solvent consumption, and handling of smaller sample volumes while providing good sensitivity and accuracy.

In this work, a novel coupled approach MicroNIR/Chemometrics based on a miniaturized and portable spectrometer is proposed for the on site detection of amphetamines (AMP) in non pretreated oral fluids. In particular, the coupling of MicroNIR with chemometrics was investigated with the aim of developing a fast and accurate approach able to perform the on-site prediction of AMP abuse. A predictive model to be used in real cases was developed by collecting specimens from volunteers and spiked samples with increasing amounts of AMP were prepared to optimize calibration. Partial Least Square-Discriminant Analysis (PLS-DA) and Partial Least Square regression (PLS) were involved to detect and quantify AMP. Results demonstrated that MicroNIR/Chemometric platform is statistically able to identify AMP abuse in simulated oral fluid samples containing, with the accuracy and sensitivity of the actual proposed official reference methods. The method was checked against false positive and true positive response and results proved to be those required for confirmatory analyses. This method would permit to simplify AMP abuse monitoring for roadside drug testing or workplace surveillance and may be of help at first aid points.

Background: Caffeine, theophylline, and theobromine are methylxanthines commonly found in coffee, tea, cola, and cocoa. Other sources may be soft drinks or energy drinks. All of them are stimulants of the nervous system and can be used for the treatment of some diseases. The three xanthines produce addiction with typical abstinence symptoms. Among young people, the consumption of energy beverages is increasing, and the growing market causes concern about the caffeine intake. To evaluate intake of methylxanthines, their accurate determination can be helpful. Methods: A simple method for their determination without sample pretreatment was applied to beverages, including coffee, tea, cola, and energy drinks. The separation was achieved by LC with UV detection. The method was validated in terms of linearity, LOD and LOQ, accuracy, and reproducibility. Results: The drinks were directly injected after a filtration, and no matrix effect was demonstrated. The procedure proved to be simple, time saving, accurate, and reproducible and may be recommended for reliable assays in routine work. The investigated samples showed a range of caffeine concentration from 100 to 3050 mg/L. Possible intake of methylxanthines from miscellaneous types of drinks was assessed considering the European Food Safety Authority Opinion on the safety of caffeine consumption. Our results were in good agreement with other authors. Conclusions: A fast and accurate method for the simultaneous determination of three xanthynes in beverages was validated. The selected strategy has proved to be fit-for-purpose by applying it to different nervous system stimulant drinks. Highlights: A simple and time saving procedure was proposed for the separation, detection, and quantitation of three methylxantynes in nervous system stimulant drinks. No sample preparation was needed, and speculation could be made about the possible intake of them from beverages.

In this paper several approaches are discussed for the direct analysis of the main sugars in different fruit juices. Refractometry, thin layer chromatography, volumetric analysis and high performance liquid chromatography with refractive index detector were tested and the results compared, discussing the advantages and disadvantages of each of them. Whereas the first method gives generically indications on the whole content of sugar and it doesn’t require any prior manipulation of the sample, thin layer chromatography is useful only for qualitative purpose, on the other hand the third method, after removal of interferences, makes possible the determination of the reducing and not reducing sugar, and the last one allows the qualitative and quantitative determination of the saccharides singularly. It’s very important to have not only knowledge about the chemical analysis of carbohydrates and their physicochemical properties, but especially how the methods can be used in product development for benefit of the public. In the wide range of options for the determination of the mono and disaccharides in beverages, the approach selected must be robust, accurate, powerful and reproducible.

Domestic biomass heating and wildfires strongly affect particulate matter (PM) concentration in the atmosphere. The individuation of alternative chemical tracers may provide a valuable tool to apportion different possible contributions to biomass burning. In this study, we used a new experimental procedure, based on high spatial resolution analyses of PM10, to assess the spatial distribution of levoglucosan (LVG) and evaluate the possible use of alternative biomass burning tracers in the Terni basin, a wide urban and industrial hot-spot of Central Italy, which includes several spatially disaggregated sources. Spatially-resolved chemical characterization of PM10 was obtained through the use of innovative samplers working in parallel in a dense monitoring network (20 sampling sites, about 1 km between each other), during wintertime. PM10 samples were analyzed for LVG, water-soluble organic carbon (WSOC) and water-soluble and insoluble fraction of 33 elements. Principal component analysis (PCA) on the obtained spatially-resolved data allowed us to identify biomass burning tracers across the polluted study area. Analyses of size-segregated PM samples showed the presence of LVG, and water-soluble Cd, Cs, K, Rb and Tl in particles with size fraction smaller than 1 μm, confirming them as tracers of combustion processes (mainly related to biomass burning). Pearson correlation coefficients demonstrated that concentrations of WSOC and water-soluble Cd, Cs, K, Rb and Tl were well correlated with the spatial variability of LVG concentration. The combined use of spatially and dimensionally resolved data was found to be particularly advantageous for the identification of alternative source tracers, which were used to reliably trace the main local biomass burning sources in the study area.

The occurrence of halogenated organic pollutants in indoor dust can be high due to the presence of textile, electronic devices, furniture, and building materials treated with these chemicals. In this explorative study, we focused on emerging organic pollutants, such as novel brominated flame retardants (nBFRs) and some perfluoroalkyl substances, together with legacy polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (BDEs) in settled dust collected in houses and workplaces such as one office and two electrotechnical and mechanical workshops. The total contribution of the investigated pollutants was lower in house and in office dusts except for few nBFRs (such as bis (2-ethylhexyl)-3,4,5,6-tetrabromo-phthalate at a concentration of 464.5 ng/g in a house and hexachlorocyclopentadienyldibromocyclooctane at 40.4 ng/g in the office), whereas in electrotechnical and mechanical workshops a high incidence of PCBs, BDEs, and nBFRs occurred (for example, BDE 209 at a concentration of 2368.0 ng/g and tetrabromobisphenol A at 32,320.1 ng/g in electrotechnical and mechanical workshops). Estimated daily intakes were also calculated, showing that domestic and occupational environments can lead to a similar contribution in terms of human exposure. The higher exposure contribution was associated to nBFRs, whose EDIs were in the range of 3968.2&ndash, 555,694.2 pg/kg bw/day. To provide a complete view about the indoor contamination, in this investigation, we also included polycyclic aromatic hydrocarbons (PAHs) and their oxygenated and nitrated derivatives. Definitely, dust collection represents a simple, fast, and cost-effective sampling and dust contamination level can be a useful indicator of environment healthiness. Besides, the presented method can be a smart tool to provide a time and money saving technique to characterize 99 pollutants thanks to a single sample treatment.

The diffusion and the recycling of electric and electronic equipment waste (WEEE) are an important issue for industrialised countries. During treatment operations of this equipment, legacy and emerging halogenated flame retardant can be released in the environment, causing great concern for their toxicity. For this purpose, a method to simultaneously determine polybrominated biphenyl ethers, hexabromocyclododecane, 10 emerging brominated flame retardants and in parallel-polychlorinated biphenyls was optimised and applied to particulate matter samples collected within a WEEE facility. In this paper, starting from a previously published method, we drastically simplified the sample preparation, reducing the overall completion time, solvent consumption, costs and achieving adequate selectivity and sensitivity for all the target compounds. The multi–analyte method was evaluated in terms of reproducibility, linearity, recovery, limits of detection, limit of quantification, and matrix effect and compared to the previous method. Quantitative analyses were carried out by gas chromatography-mass spectrometry in negative chemical ionisation, using matrix-matched calibration curves. The reliability and robustness of the method were demonstrated through the analysis of certified and reference compounds in Standard Reference Material 2585. The most abundant compounds in particulate matter samples, collected in an electric and electronic equipment waste treatment plant, were BDE 47 (3.9 ng m−3), CB 138 (2.7 ng m−3), and 1,2-bis(2,4,6-tribromophenoxy) ethane (4.2 ng m−3).

Oxygenated polycyclic aromatic hydrocarbons are toxicologically relevant pollutants to both humans and environment, identified in urban aerosols, coming from both direct emissions, and heterogeneous reactions in atmosphere. In this paper, 7H-benz[ de ]anthracene-7-one, benz[ a ]anthracene-7,12-dione and benzo[ a ]fluoren-11-one concentrations were studied in size-segregated atmospheric particles collected in Rome, Italy, over one year. The relevant concentration of oxy-PAHs (as sum of three compounds), found in the warmer season (2.1 ng m −3 ), was higher than both polycyclic aromatic hydrocarbon (as sum of fourteen PAHs) and their nitro-derivative (as sum of ten nitro-PAHs) concentrations in the same samples (respectively 1.31 and 0.07 ng m −3 ). The warmer season is also characterized by oxy-PAH higher concentration in the ultrafine fraction of the aerosol (0.74 ng m −3 ) when compared to the same fraction in winter and intermediate seasons (0.22 and 0.09 ng m −3 , respectively). Due to oxy-PAH major distribution in particles capable to reach the human target organs, these results suggest an important role for oxy-PAHs in the air direct-acting mutagenicity, in urban sites, in summer.

An effective analytical method for the screening of mycotoxins, in indoor/outdoor airborne particulate matter, was developed and method performance data are presented. Mycotoxins are natural compounds produced, in particular conditions, as secondary metabolites by filamentous fungi and moulds, and, after their production, they can be transported far from their source. To simulate real samples, an urban dust (reference material 1649a) free from mycotoxins was used as matrix and spiked by the most common mycotoxins, chosen on the basis of studies carried out previously in other real matrices: deoxynivalenol, aflatoxin B1, ochratoxin A, T-2 toxin, zearalenone and sterigmatocystin. The analytical method was optimised and structured in four successive steps: (1) accelerated solvent extraction of the (spiked) analytes from matrix, (2) solid-phase purification (SPE) of the previous extract, (3) pre-concentration of the eluates from SPE and (4) analysis of the concentrated eluates by high performance liquid chrom...

Abstract In Kluyveromyces lactis yeast, OCH1 encodes for the a-1,6-mannosyltrasferase that adds the initial a-1,6-mannose to the outer-chains of Nglycoproteins. Kloch1-1 mutant cells showed altered calcium homeostasis and endoplasmic reticulum (ER) stress. Since ER plays a major role in lipid biosynthesis and lipid droplet (LD) formation, herein the impact of Och1p depletion on lipid homeostasis was investigated. Transcriptional profiles of genes involved in biosynthesis of fatty acids, their amount and composition changed in mutant cells. An increased amount of ergosterol was determined in these cells. Enhanced transcription of genes involved in both synthesis and mobilization of LDs was also found in Kloch1-1 cells, accompanied by a reduced amount of LDs. We provide evidence that ER alterations, determined by protein misfolding as a result of reduced N-glycosylation, induced altered lipid homeostasis in Kloch1-1 cells. Chemical chaperone 4-phenyl butyrate (4-PBA) slightly alleviated the LD phenotype in cells depleted of Och1p. Remarkably, complete suppression of ER stress, via increased expression of plasma membrane calcium channel subunit Mid1, fully restored lipid homeostasis in mutant cells. To further reinforce this finding, low numbers of LDs were observed in wild type cells when ER stress was triggered by DTT treatment.

Fosetyl-aluminum is a synthetic fungicide administered to plants especially to prevent diseases caused by the members of the Peronosporales and severalPhytophthoraspecies. Herein, we present a selective liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to analyze residues of fosetyl-A1 in air particulate matter. This study was performed in perspective of an exposure assessment of this substance of health concern in environments where high levels of fosetly-Al, relatively to airborne particulate matter, can be found after spraying it. The cleanup procedure of the analyte, from sampled filters of atmospheric particulate matter, was optimized using a Strata X solid-phase extraction cartridge, after accelerated extraction by using water. The chromatographic separation was achieved using a polymeric column based on hydrophilic interaction in step elution with water/acetonitrile, whereas the mass spectrometric detection was performed in negative electrospray ionization. The proposed method resulted to be a simple, fast, and suitable method for confirmation purposes.

Organic component of airborne particulate matter originates from both natural and anthropogenic sources whose contributions can be identified through the analysis of chemical markers. The validation of analytical methods for analysis of compounds used as chemical markers is of great importance especially if they must be determined in rather complex matrices. Currently, standard reference materials (SRM) with certified values for all those analytes are not available. In this paper, we report a method for the simultaneous determination of levoglucosan and xylitol as tracers for biomass burning emissions, and arabitol, mannitol and ergosterol as biomarkers for airborne fungi in SRM 1649a, by GC/MS. Their quantitative analysis in SRM 1649a was carried out using both internal standard calibration curves and standard addition method. A matrix effect was observed for all analytes, minor for levoglucosan and major for polyols and ergosterol. The results related to levoglucosan around 160 μg g−1 agreed with those reported by other authors, while no comparison was possible for xylitol (120 μg g−1), arabitol (15 μg g−1), mannitol (18 μg g−1), and ergosterol (0.5 μg g−1). The analytical method used for SRM 1649a was also applied to PM10 samples collected in Rome during four seasonal sampling campaigns. The ratios between annual analyte concentrations in PM10 samples and in SRM 1649a were of the same order of magnitude although particulate matter samples analyzed were collected in two different sites and periods.

This work is part of an extensive research project aimed at the determination and characterization of bioaerosol with a multidisciplinary approach. In this context, one of the main objectives of the project has been the development of a comprehensive analytical method for the determination of different chemical biomarkers of the bioaerosol, by liquid chromatography coupled with tandem mass spectrometry. The following biomarkers have been considered, and correlated to specific components of bioaerosol as unambiguous indicators: • ergosterol → fungal components • chlorophylls, phytosterols (stigmasterol and b-sitosterol), α-tocoferol → vegetable cells and algae • cholesterol → animal cells, vegetable cells and algae. • dipicolinic acid → bacterial spores • muramic and meso-2,6-diaminopimelic acid → bacterial cells To verify the method, to find diagnostic ratios and to calculate the appropriate conversion factors, fungal spores, bacterial cells and spores, and algae of known species, commonly airborne, were analysed. The material was subjected to freezing and de-freezing cycles, followed by extraction, hydrolysis and purification of the biomarkers. The chromatographic separation of the bacterial biomarkers was achieved by using a polymeric column, based on Hydrophilic Liquid Interaction with the electrospray ionization mass spectrometric detection, whereas sterols and chlorophylls were separated by a reversed phase column, coupled to atmospheric pressure chemical ionization – tandem mass spectrometer. The optimized method was applied to environmental particulate matter sampled in an outdoor site. Bacterial and fungal content was compared to the results obtained from the classical direct viable counting method in the sampled particulate matter.

Mycotoxins exposure by inhalation and/or dermal contact is possible in different branches of industry especially where heavily dusty settings are present and the handling of dusty commodities is performed. This study aims to explore the validity of the biomonitoring as a tool to investigate the intake of mycotoxins in a population of workers operating in an Italian feed plant. Serum samples were collected for the determination of aflatoxins B1 (AFB1), AFB1-Lysine adduct and ochratoxin A (OTA). A method based on liquid&ndash, liquid extraction coupled with high resolution mass spectrometry determination was developed and fully validated. For AFB1, a high number of non-detected samples (90%) was found and no statistical difference was observed comparing workers and control group. None of the analyzed samples showed the presence of AFB1-Lysine adduct. For OTA, the 100% of the analyzed samples was positive with a 33% of the samples showing a concentration higher than the limit of quantification (LOQ), but no statistical difference was highlighted between the average levels of exposed and control groups. In conclusion, the presence of AFB1 and OTA in serum cannot be attributable to occupational exposure.

A liquid chromatography–tandem mass spectrometry analytical procedure has been developed for the detection and quantitative determination of underivatized amino acids at low concentrations in a Standard Reference Material—urban dust. In order to minimize interferences of other compounds, an accelerated solvent extraction followed by a solid phase extraction on two different cartridges was applied prior to LC–MS–MS. Fourteen amino acids were separated by high resolution liquid chromatography, detected and quantified by multiple reaction monitoring on a triple quadrupole. The proposed methodology has been applied for the first time on Standard Reference Material 1649a (urban dust) from the National Institute of Standards and Technology, that does not report certification values for these compounds. This methodology avoids the derivatization step and allows the amino acid quantification in a complex matrix, such as that of atmospheric particulate matter, and represent a good method suitable to analyze this class of compounds in atmospheric aerosol. The selected strategy demonstrated to be fit-for-purpose, by applying it to a real atmospheric sample with the aim to verify the efficacy of the study and to provide information about the organic matter content.

The present paper deals with identification of hydrocarbon sources in contaminated soil of three tank farms located in north, central and south Italy. Parent polycyclic aromatic hydrocarbons (naphthalene through benzo(g,h,i)perylene) and n-alkanes (n-C10 through n-C36) were determined. The study of source assignments was carried out by their distribution patterns, their diagnostic ratios, and determination of less-commonly used markers. The data show anthropogenic and biogenic origin of n-alkanes and petrogenic and pyrolytic sources of PAHs. Multiple sources of hydrocarbons, that were not considered in the preliminary environmental assessments, are identified. The application of a multi-criteria approach allows to locate petroleum pollution sources that affect the study sites. The results demonstrate that the application of a multi-criteria approach for source identification is a key point to assess environmental damage and prove that an accurate study of source identification has to be performed. The suggested methodology is a useful tool to manage contaminated sites and to plan appropriate interventions of clean up.

Airborne microorganisms, as bacteria and fungi, are ubiquitous components of the atmospheric aerosol particles. In this paper, we report a method for the simultaneous extraction, purification, separation, identification and quantification of ergosterol, mannitol and arabitol as biomarkers of fungal spores in bioaerosol particles. After sampling by a low volume sampler, filters were spiked with mannitol-13C and dehydrocholesterol as internal standards. Samples were then extracted by accelerated solvent extraction using pure ethanol. The extract was then passed through an amino cartridge and divided in two parts: the apolar fraction, released from the cartridge, was subjected to liquid liquid extraction (by n-hexane), while polar compounds, retained by the cartridge, were eluted by a mixture of methanol–water. The two fractions were joined and analyzed by HPLC equipped with two different columns in series, and coupled to a triple-quadrupole mass spectrometer with Atmospheric Pressure Chemical Ionization source. In addition, the same fractions were analyzed, after derivatization, by GC–MS. The results obtained by the two techniques were finally compared, showing good agreement between them. Last, the contents of the three biomarkers have been estimated in three atmospheric samples collected in a suburban/rural site and, using literature conversion factors, correlated to fungal biomass.

A novel multi-analyte method for the simultaneous determination of 46 compounds of environmental concern, most of them belonging to the category of persistent organic pollutants, was developed using high-performance liquid chromatography and the results were compared to those obtained by gas chromatography. This study was performed in perspective of a cumulative exposure assessment of substances of health concern in environments where high levels, relatively to airborne particulate matter, can be found. The target compounds included polychlorinated biphenyls, brominated flame–retardants and derivatives of polycyclic aromatic hydrocarbons. The multi-analyte method was evaluated in air particulate matter in terms of reproducibility, linearity, recovery, limits of detection and quantification and matrix effect. The recovery was above 70% for all the analytes, whereas limits of quantification ranged between 23 and 390 pg∙m−3 in liquid chromatography and less than ten times in gas chromatography–mass spectrometry. Matrix effect was generally negligible for both the techniques, except the case of the detection of oxygenated derivatives of polycyclic aromatic hydrocarbons by gas chromatography. In order to demonstrate the efficacy and to assess the method performances (accuracy and precision), both the techniques were applied to standard reference materials, and the results were compared, discussing their advantages and disadvantages. The method was finally applied to a real sample of indoor airborne particulate matter with aerodynamic diameter ≤4 μm (PM4). We demonstrated that liquid chromatography was the only technique able to analyse the 46 compounds, including thermally degradable ones, with a single chromatographic run without derivatisation steps. On the other hand, gas chromatography still presents higher sensitivity for the detection of some of the investigated compounds. This study can be considered only explorative and further improvements can be expected with new-generation LC-MS instruments (10–100 times more sensitive).

Fungal spores are the dominant biological component of air. Although ubiquitous in outdoor air, they are scarcely measured due to the inadequacy of measurement methods. The use of biomarkers as tools for the determination of fungal contribution to bioaerosol has often been suggested, and ergosterol, arabitol and mannitol have been associated to fungal spores as tracers. In the present paper, the fungal component of aerosol was studied at suburban/rural and at urban sites. Ergosterol, arabitol, and mannitol contents in airborne particulate matter, even at different sizes, were determined. Literature conversion factors and calculated conversion factors correlating ergosterol, arabitol, and mannitol masses to fungi mass were applied and compared to each other. The obtained fungal spore concentrations were different depending on the marker utilized both with the conversion factors found in literature and the calculated ones. Size-segregated marker distribution suggested different sources for the three tracers indicating ergosterol as the only reliable biomarker at our latitudes. The fungal spore concentrations were higher at the suburban/rural location and respectively inversely and directly proportional to temperature and relative humidity.

Avermectines are antiparasitic agents widely used as veterinary drugs for food producing animals. The European Community, due to their side effects, limited the use of these molecules establishing maximum residue limits (MRLs) in some foods. A validated qualitative and quantitative high performance liquid chromatography method with fluorescence detection (HPLC-FL) is presented for the simultaneous determination of ivermectin (IVM), abemectin (ABA), moxidectin (MOX), eprinomectin (EPR), doramectin (DOR) and emamectin (EMA) in foodstuffs (muscle, eggs and milk). Samples were extracted with acetonitrile, purified with liquid-liquid extraction (LLE), and analysed by HLPC-FL previous derivatization with trifluoroacetic anhydride (TFAA) in presence of 1-methyl-imidazole (MI) and acetic acid. To date, the presented method is the first validated for the matrix eggs, and in accordance with the requirements set by Commission Decision 2002/657/EC. Recoveries of the methods, calculated spiking the samples in the range 5.0–100.0 μg kg −1 , were 64–83% for muscle, 65–89% for milk and 63–84% for eggs. The precision (CV) ranged between 9.2 and 17.1% for muscle, 9.9 and 16.6% for milk and from 9.4 to 17.4% for eggs. Linearity for the six analytes was calculated from 5.0 to 200.0 μg kg −1 . The main advantages of the presented method are its rapidity, the specificity, the good precision and recovery that make it very suitable to the detection and determination of avermectines.

Size-segregated atmospheric particles were collected in Rome, Italy, using a low-pressure impactor. Twelve sampling campaigns were conducted under different meteorological conditions over a whole year covering 155 not consecutive days. The samples were analyzed for polycyclic aromatic hydrocarbons (PAHs) and their nitro- and methyl- derivatives known for their toxicity. An assessment of the carcinogenic potency of the particles known to penetrate into lungs, liver, heart and nervous system was performed. The distribution of the classes of compounds was unimodal and centrated at 0.4 μm size fraction for PAHs and bimodal and centrated at 0.1 and 0.4 μm for methyl- and nitro- derivatives. The 18% of toxic organic compounds we analyzed was distributed into the ultrafine fraction (PM0.1) and 76% in the fine fraction; but substituted PAH distribution in the ultrafine particles shifted toward higher values during warm periods. In July, the 50% of the total nitro-PAHs was found in PM0.1 and an average of 42% of the total methyl-PAHs was found in the same fraction in summer and intermediate seasons. An evaluation of the potential toxicity of the measured compounds was tentatively assessed based on Potency Equivalency Factors (PEF).

Nandrolone (19-nortestosterone) is an androgenic anabolic steroid illegally used as a growth-promoting agent in animal breeding and as a performance enhancer in athletics. Therefore, its use was officially banned in 1974 by the Medical Commission of the International Olympic Committee (IOC). Following nandrolone administration, the main metabolites in humans are 19-norandrosterone, 19-norethiocolanolone and 19-norepiandrosterone, and their presence in urine is the basis of detecting its abuse. The present work was undertaken to determine, in human urine, nandrolone metabolites (phase I and phase II) by developing and comparing multiresidue liquid chromatography/tandem mass spectrometry (LC/MS/MS) and gas chromatography/mass spectrometry (GC/MS) methods. A double extraction by solid-phase extraction (SPE) was necessary for the complete elimination of the interfering compounds. The proposed methods were also tested on a real positive sample, and they allow us to determine the conjugated/free fractions ratio reducing the risk of false positive or misleading results and they should allow laboratories involved in doping control analysis to monitor the illegal use of steroids. The advantages of LC/MS/MS over GC/MS (which is the technique mainly used) include the elimination of the hydrolysis and derivatization steps: it is known that during enzymatic hydrolysis several steroids can be converted into related compounds and deconjugation is not always 100% effective. The validation parameters for the two methods were similar (limit of quantification (LOQ)

Resveratrol (3,5,4′-trihydroxystilbene) is a phytoalexin that belongs to the group of stilbenes, known to occur in grapes and consequently in grape products. Its presence in wine is an important qualitative parameter because of the several beneficial effects on human health. The aim of this work is the development of a high-performance liquid chromatographic (HPLC) method for the determination of trans resveratrol in wines, and comparisons between the results obtained by different detection techniques: UV-vis spectroscopy, fluorescence spectroscopy and mass spectrometry. Resveratrol is analysed on a C-18 column using gradient elution. The method permits direct injection of sample, revealing to be time-saving, overcoming the need of sample pre-treatment steps. Detection limits were 154.8 ng mL−1 by HPLC-UV, 118.0 ng mL−1 by HPLC-FL and 48.0 ng mL−1 by HPLC-MS. Trans resveratrol has been then quantified in a group of 52 wines derived from different Italian regions, cultivars and winemaking technologies by HPLC-UV.

Boldenone is an androgenic anabolic steroid (AAS) intensively used for growth promoting purposes in animals destined for meat production and as a performance enhancer in athletics. Therefore its use is officially banned either in animals intended for consumption or in humans. Because most anabolic steroids are completely metabolized and usually no parent steroid is excreted, metabolite identification is crucial to detect the illegal use of anabolic steroids either in humans or in livestock. The aim of this work is the investigation of 17-boldenone and its main metabolites, 17-sulphate, 17-glucuronide, 5-androst-1-en17-ol-3-one and 17-boldenone, in human and bovine urine developing a multiresidue analysis. After solid phase extraction of urine samples, detection is carried out by high performance liquid chromatography–tandem mass spectrometry in multiple reaction monitoring. The average recovery for all the investigated compounds is above 70%. The developed method is very easy, quick and highly specific. Linearity, precision, decision limit and detection capability were also evaluated. © 2005 Elsevier B.V. All rights reserved.

An analytical procedure for the determination of natural steroids and their conjugates was developed and applied to bovine and human urine. Fifteen compounds (free and conjugated) after extraction by solid-phase were identified by liquid chromatography–tandem mass spectrometry (LC–MS–MS) in multiple reaction monitoring (MRM). Free and conjugated steroids were detected all together in the same chromatographic analysis respectively in positive and negative ionization. This procedure reveals to be time-saving, because it overcomes the need of further sample pre-treatment steps, such as enzymatic hydrolysis and derivatisation. The recovery for each compound was around 80%. Limit of detection of each steroid free or conjugated has been evaluated.

In this work several compounds were detected in wines by HPLC-tandem mass spectrometry. In particular cinnamic and benzoic acids, tyrosol, apigenin-7-glucoside and luteolin-7-glucoside were identified and quantified in Italian wines. Red wines show bigger amount of cinnamic and benzoic acids than white wines. tyrosol is in bigger amount with respect to two flavones: luteolin-7-glucoside and apigenin-7-glucoside. These last two flavones are only in some wine, but it can be important to detect the presence of different substances in small amount to be able to characterize a wine.

This paper describes a method for analysing some acids of milk whey by Capillary Zone Electrophoresis. After eliminating the whey proteins by ultrafiltration, the whey underwent electrophoretic separation in the presence of anodic electroosmotic flow. The following analytes were detected: citric, orotic, uric, and hippuric acids. A procedure is described for sample preparation and the operating conditions for electrophoretic capillary separation established. Finally, orotic acid is quantitatively determined.

A liquid chromatographic–tandem mass spectrometric (LC–MS–MS) method has been developed for the determination of trenbolone in bovine urine and serum. The aim was a control of the misuse of trenbolone in food-producing animals. The procedure involved, in both cases, a preliminary solid-phase clean-up followed by a liquid–liquid extraction for urine samples after a preliminary enzymatic hydrolysis. The extracts have been directly analysed by reversed-phase LC–MS–MS in selected reaction monitoring (SRM), acquiring two diagnostic product ions from the chosen precursor [M+H] + . The procedures were validated across the concentration range of 1–1500 ng/ml. The linearity, the inter- and intra-day accuracy and precision have been determined. The procedure was specific and the accuracy values were better than 20% at the limit of quantitation of spiked samples. The limit of quantification (LOQ) and the limit of detection (LOD) were, respectively, 1 ng/ml and 350 pg/ml for urine and serum. According to the draft, SANCO/1805/2000, we determined the decision limit CC α and the detection capability CC β . The recovery values for urine ranged from 87 to 128%, and for plasma the recovery was 70±4%. The procedure proved to be simple and suitable for routine and confirmatory purposes such as those developed for residue studies.

Corticosteroids can be illegally administered to cattle as growth promoting agents to improve meat production. We developed a liquid chromatography-atmospheric pressure ionization mass spectrometry-mass spectrometry (LC-MS-MS) method able to identify and quantify flumethasone, one of the most potent fluorinated synthetic corticosteroid, in serum and urine from treated calves. The analyte was purified from urine (conjugated and free, following enzymatic hydrolysis) and from serum by C18 solid-phase and liquid-liquid extractions, then analyzed by LC-MS-MS monitoring the product ions of an abundant precursor (SRM in negative ionization mode). Results on flumethasone residues in biological fluids in three calves treated at different levels are presented. This method allowed the detection of flumethasone in bovine urine and serum at the 30-pg/ml level.

The aim of this work was to analyze by HPLC-MS-MS natural biophenolic compounds in complex natural samples like olives and olive oils. Eleven benzoic and cinnamic acids, vanillin and other eight bi...

Some citrus essential oils were analyzed by HPLC with both microbore and standard columns in reversed and normal phase. Volatile and non-volatile fraction were investigated. In the non-volatile fraction some coumarins have been identified. Fractions are detected spectrophotometrically at 220 nm and 320 nm before and after evaporation of samples. Some components were also identified by LC/MS.

A rapid and efficient capillary zone electrophoresis method was developed for Quality Control analysis of pharmaceutical preparations containing antihistamines, decongestants and anticholinergic remedies: chlorpheniramine, diphenhydramine, ephedrine, isopropamide, pheniramine. These analytes were separated in cationic form optimizing the experimental conditions in 60 mM tetraborate buffer pH = 9.2 as a BGE (Background Electrolyte) on a Beckman P/ACE System MDQ instrument. The effective capillary length was 48 cm, I.D. = 75 μm, the applied voltage 15 kV, and the temperature 25 °C. Detection was performed by a DAD (Diode Array Detector) at 210 nm. Separation time was less than 8 min. After experimental conditions optimization, the proposed method was validated. Precision of migration time (tm) ranging from 0.19% to 0.29% and corrected peak area (Ac) from 2.54% to 3.68%. The linearity of detector response was tested in the range 5–40 μg ml−1 obtaining the 0.9962 ≤ r2 ≤ 0.9982. LOD and LOQ, accuracy (recovery) and ruggedness were evaluated for each analyte demonstrating the good reliability of the method. Analysis of the pharmaceutical real sample was performed.