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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.

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.

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.

Speciation studies are based on fundamental models that relate the properties of biomimetic coordination compounds to the stability of the complexes. In addition to the classic approach based on solution studies, solid state properties have been recently proposed as supporting tools to understand the bioavailability of the involved metal. A ten-year long systematic study of several different complexes of imidazole substituted ligands with transition metal ions led our group to the definition of a model based on experimental evidences. This model revealed to be a useful tool to predict the stability of such coordination complexes and is based on the induced behavior under thermal stress. Several different solid state complexes were characterized by Thermally Induced Evolved Gas Analysis by Mass Spectrometry (TI-EGA-MS). This hyphenated technique provides fundamental information to determine the solid state properties and to create a model that relates stability to coordination. In this research, the model resulting from our ten-year long systematic study of complexes of transition metal ions with imidazole substituted ligands is described. In view of a systematic addition of information, new complexes of Cu(II), Zn(II), or Cd(II) with 2-propyl-4,5-imidazoledicarboxylic acid were precipitated, characterized, and studied by means of Thermally Induced Evolved Gas Analysis performed by mass spectrometry (TI-EGA-MS). The hyphenated approach was applied to enrich the information related to thermally induced steps, to confirm the supposed decomposition mechanism, and to determine the thermal stability of the studied complexes. Results, again, allowed supporting the theory that only two main characteristic and common thermally induced decomposition behaviors join the imidazole substituted complexes studied by our group. These two behaviors could be considered as typical trends and the model allowed to predict coordination behavior and to provide speciation information.

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.

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.

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...

In this paper, we report a simple method for the determination of quercetin, a typical antioxidant compound ubiquitously present in vegetable food products including grapes and wine. To this aim, 12 Italian wines were analyzed and the amount of quercetin was determined by high-performance liquid chromatography with ultraviolet detector. To reduce the interferences, a minimal sample preparation was performed on solid phase cartridge, then the samples were eluted through a narrow bore C18 column under gradient elution in less than 25 min. Nine wines were red and three white. The amount of quercetin ranged from 0.5 to 4.3 mg L−1. The method was evaluated in terms of reproducibility, linearity, recovery, and limits of detection and quantification, and it resulted relatively fast, economical, and suitable for those laboratories involved in routine analysis. In addition, the possibility to determine the content of quercetin derivates, as free quercetin, after acidic hydrolysis was investigated. Despite the limited number of analyzed samples, the results from this explorative study are in agreement to those from other surveys. Moreover, due to the problems arising from hydrolysis, further research are needed for determining each single quercetin derivate in wine, in order to better characterize its active compounds.

Coccidiostats are synthetic drugs administered to animals, especially to poultry, to cure coccidiosis. In this paper, we present a selective liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to analyze residues of five synthetic coccidiostats in eggs: clazuril, diclazuril, robenidine, nicarbazin, toltrazuril and its two metabolites. The extraction efficiency was evaluated by testing several solvents, pH, different volumes and time of extraction. The clean-up procedures were optimized using different solid phase extraction cartridges and different eluants. The chromatographic separation was achieved in reversed phase using a gradient of 0.1% formic acid in water and acetonitrile, whereas the MS detection was performed in negative electrospray ionization (ESI) for all the analytes, except for the robenidine. The developed method has been validated according to Commission Decision 2002/657/CE. The validation parameters, as linearity, precision, recovery, specificity, decision limit (CCα), detection capability (CCβ), and robustness have been determined. The proposed method resulted simple, fast, and suitable for screening and 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.

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.

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.

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.

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...

In this work hydroxycinnamic acid esters of tartaric acid (mono-p-coumaroyl tartaric acid, monocaffeoyl tartaric acid, monoferuloyl tartaric acid esters) were separated, identified and quantified in several red wines by high performance liquid chromatography coupled with tandem mass spectrometry. The method features direct analysis of wines with no preparation and analysis of grape juice with minimum sample manipulation. For the identification was used both a triple quadrupole and a quadrupole–Tof, while for the quantification a triple quadrupole. At the same time two impurities, due to interfering substances coming from methanol (stored in bottles with polypropylene tops without Teflon protection), were characterised. These impurities had the same molecular weight as the investigated compounds.

Stilbenes from grapes and wines play a central role in the human diet because of their antioxidant, antimutagenic and anticarcinogenic properties. We describe a method for the direct determination of some stilbenes (cis- and trans-resveratrol, cis- and trans-resveratrol glucoside, cis- and trans-piceatannol, and cis- and trans-piceatannol glucoside) in wine by high-performance liquid chromatography/mass spectrometry using a triple quadrupole (QqQ) mass spectrometer, in multiple reaction monitoring (MRM) mode, acquiring two diagnostic product ions from the chosen precursor. All the target analytes were separated on a C-18 column using gradient elution, in a single run. Electrospray ionization (ESI) in negative ion mode gives higher sensitivity for all the target compounds than atmospheric pressure chemical ionization (APCI). For the identification of piceatannol glucoside (astringin), because of the lack of a suitable standard, an HPLC/TOFMS method was used. The method permits direct injection of samples and it is time-saving, removing the need for sample pre-treatment. The detection limits were 48.0 ng mL(-1) for cis- and trans-resveratrol and for cis- and trans-resveratrol glucoside, and 50.0 ng mL(-1) for cis- and trans-piceatannol. The procedure proved to be simple and suitable for routine and confirmatory purposes. A total of 19 red and 3 white Italian wines were analyzed and differences in the stilbene composition were found among these samples.

Stanozolol is an anabolic steroid illicitly used for growth promoting purposes in animal production. For reasons of public health the use of anabolic steroids as growth promoters is officially banned in Europe in animals intended for consumption. The aim of this work is the investigation of stanozolol metabolites, 3′-hydroxystanozolol, 4β- hydroxystanozolol and 16β- hydroxystanozolol in urine samples after stanozolol administration to a young calf. After solid phase extraction of urine samples, detection is carried out by HPLC-MS-MS multiple reaction monitoring. Average recovery for the three metabolites is 80%. The method is highly specific and has been validated in terms of linearity, inter and intra day precision. In addition the decision limit CCα and the detection capability CCβ have been determined.

Biophenolic compounds constitute a complex mixture in olives and derived products. In this work, ion-spray tandem mass spectrometry coupled with liquid chromatography is applied to the identification and quantification of biophenols present in complex matrices such as olives and vegetation waters. The extraction and purification method for the biophenol present in both the matrices is reported. Using this approach the main biophenols found, in three different variety of olives, were: tyrosol, hydroxytyrosol, oleoside methyl ester, oleoside dimethyl ester, and oleuropein. In vegetation water the biophenols are present in lower amounts than in olives.

The metabolism of oxabolone cipionate, 17-(3-cyclopentyl-1-oxopropoxy)-4-hydroxyestr-4-en-3-one, a synthetic anabolic steroid, was investigated in man, the cumulative urinary excretion and the metabolism of the compounds being studied by GC-MS in both electron impact and chemical ionization modes. After administration by injection to volunteers, five different metabolities were detected in urine. The metabolites and the parent compound were detected in urine up to a week after administration.

A method for the extraction and separation of the non-volatile phenolic acids of wine is described. The extracts are analysed by HPLC with a microbe column and UV detection. The free phenolic acids and depsides in different wine samples were identified and determined.