Determination and risk assessment of phthalates in face masks. An Italian study.

Serious human health concerns have been recently raised from daily use of face masks, due to the possible presence of hazardous compounds as the phthalic acid esters (PAEs). In this study, the content of 11 PAEs in 35 commercial masks was assessed by applying a specific and accurate method, using Gas Chromatography/Mass Spectrometry. Surgical, FFP2 and non-surgical models, for both adults and children were collected from the Italian market. Analyses showed that four of the target analytes were detected in all tested samples with median total concentrations ranging between 23.6 mg/kg and 54.3 mg/kg. Results obtained from the experimental analysis were used in the risk assessment studies carried out for both carcinogenic and non-carcinogenic effects. Doses of exposure (Dexp) of PAEs ranged from 6.43 × 10-5 mg/kg bw/day to 1.43 × 10-2 mg/kg bw/day. Cumulative risk assessment was performed for non-carcinogenic and carcinogenic effects. No potential risk was found for non-carcinogenic effects, yet the 20% of the mask samples showed potential carcinogenic effects for humans. A refined exposure assessment was performed showing no risk for carcinogenic effects. This paper presents a risk assessment approach for the identification of potential risks associated to the use of face masks.

Quantification of preservatives in tattoo and permanent make-up inks in the frame of the new requirements under the REACH Regulation.

BACKGROUND: According to the REACH (Registration, Evaluation, Authorisation and Restriction of Chemicals) restriction, tattoo and permanent make-up (PMU) inks placed on the European Union market after January 4, 2022, shall not contain methylisothiazolinone, benzisothiazolinone (BIT), octylisothiazolinone (OIT), or other skin sensitizers in concentrations of 10 mg/kg or higher and phenoxyethanol (PE) or other eye irritants or damaging substances in concentrations of 100 mg/kg or higher. In addition, preservatives and other substances enlisted in Annex II to Cosmetic Product Regulation shall not be present in concentrations of 0.5 mg/kg or higher. OBJECTIVES: This study aims to quantify 14 preservatives in 99 tattoo and 39 PMU inks from the Italian market and presents a comparison with concentration limits set by the REACH restriction. METHODS: Inks were analysed by applying validated analytical methods based on liquid chromatography techniques. RESULTS: About 24.0%, 15.2% and 1.5% of the overall samples contained BIT, PE and OIT, respectively, at concentrations exceeding REACH concentration limits. The number of noncompliant tattoo inks (49.5%) would be significantly greater than that of the PMU inks (17.9%). CONCLUSIONS: About 40.6% of the samples would be noncompliant with the restriction for the presence of preservatives above the permitted level. Additional concentration limits will apply to skin sensitizing preservatives for proper labelling of inks under CLP (Classification, Labelling and Packaging) Regulation.

Monitoring of alcohol-based hand rubs in SARS-CoV-2 prevention by HS-GC/MS and electrochemical biosensor: A survey of commercial samples.

Alcohol-based hand rubs (ABHRs) have found large diffusion during the Severe Acute Respiratory Syndrome Coronavirus 2, SARS-CoV-2, thus becoming the most widespread means for hand hygiene. Whereby, it is fundamental to assess the alignment of commercial ABHRs to the indications provided by the principal health agencies regarding alcohol content and possible impurities. In this work, a novel improvement of previous existent methods for the determination of alcohol content in such products was reported. In particular, two alternative sensitive and reproducible methods, such as an electrochemical screen-printed based enzymatic (alcohol oxidase) biosensor and a Headspace Gas Chromatography coupled with Mass Spectrometry (HS-GC/MS) were proposed. The electrochemical device represents a rapid, low-cost and accurate fraud screening method for alcohol-based hand rubs. The second technique confirms, identifies and simultaneously determines ethyl alcohol, isopropyl alcohol, n-propyl alcohol and methyl alcohol, improving their extraction through acidification in the sample pre-treatment step. The developed specific HS-GC/MS method was in-house validated according to ISO/IEC 17025 requirements. Analytical parameters such as limit of detection (LoD 0.13%v/v - 0.17%v/v), limit of quantification (LoQ 0.44% v/v - 0.57% v/v), inter-day repeatability (RSDR 2.1-10.7%) and recovery (80-110%) were assessed. The relative expanded uncertainties range (between 0.1%v/v and 3.4%v/v) for all the analytes were evaluated. Results obtained using the different analytical approaches were compared and indicated that the two data sets were comparable (median; HS-GC/MS, 56%v/v; electrochemical biosensor, 62%v/v) and were not statistically different (one-way ANOVA test; p = 0.062). In addition, a good correlation (95%) was found. This study noticed that only 39% of the tested hand sanitiser products had the recommended average alcohol content, thus highlighting the need for analytical controls on this type of products.

Associations among exposure to methylmercury, reduced Reelin expression, and gender in the cerebellum of developing mice.

Genetic risk factors acting during pregnancy or early after birth have been proposed to account for the exponential increase of autism diagnoses in the past 20 years. In particular, a potential link with exposure to environmental mercury has been suggested. Male sex constitutes a second risk factor for autism. A third potential genetic risk factor is decreased Reelin expression. Male heterozygous reeler (rl(+/-)) mice show an autism-like phenotype, including Purkinje cells (PCs) loss and behavioral rigidity. We evaluated the complex interactions between 3 risk factors, i.e. genetic status, sex, and exposure to methylmercury (MeHg), in rl(+/-) mice. Mice were exposed to MeHg during the prenatal and early postnatal period, either at a subtoxic dose (2 ppm in Dams' drinking water), or at a toxic dose (6 ppm Dams' drinking water), based on observations in other rodent species and mice strains. We show that: (a) 2 ppm MeHg does not cause PCs loss in the different animal groups, and does not enhance PCs loss in rl(+/-) males; consistent with a lack of overt neurotoxicity, 2 ppm MeHg per se does not cause behavioral alterations (separation-induced ultrasonic calls in newborns, or sociability and social preference in adults); (b) in stark contrast, 6 ppm MeHg causes a dramatic reduction of PCs number in all groups, irrespective of genotype and sex. Cytochrome C release from mitochondria of PCs is enhanced in 6 ppm MeHg-exposed groups, with a concomitant increase of µ-calpain active subunit. At the behavioral level, 6 ppm MeHg exposure strongly increases ultrasonic vocalizations in all animal groups. Notably, 6 ppm MeHg significantly decreases sociability in rl(+/-) male mice, while the 2 ppm group does not show such as decrease. At a subtoxic dose, MeHg does not enhance the autism-like phenotype of male rl(+/-) mice. At the higher MeHg dose, the scenario is more complex, with some "autism-like" features (loss of sociability, preference for sameness) being evidently affected only in rl(+/-) males, while other neuropathological and behavioral parameters being altered in all groups, independently from genotype and sex. Mitochondrial abnormalities appear to play a crucial role in the observed effects.

Exosome release and low pH belong to a framework of resistance of human melanoma cells to cisplatin.

Intrinsic resistance to cytotoxic drugs has been a main issue in cancer therapy for decades. Microenvironmental acidity is a simple while highly efficient mechanism of chemoresistance, exploited through impairment of drug delivery. The latter is achieved by extracellular protonation and/or sequestration into acidic vesicles. This study investigates the importance of extracellular acidosis and nanovesicle (exosome) release in the resistance of human tumour cell to cisplatin (CisPt); in parallel to proton pump inhibitors (PPI) ability of interfering with these tumour cell features. The results showed that CisPt uptake by human tumour cells was markedly impaired by low pH conditions. Moreover, exosomes purified from supernatants of these cell cultures contained various amounts of CisPt, which correlated to the pH conditions of the culture medium. HPLC-Q-ICP-MS analysis revealed that exosome purified from tumour cell culture supernatants contained CisPt in its native form. PPI pre-treatment increased cellular uptake of CisPt, as compared to untreated cells, in an acidic-depend manner. Furthermore, it induced a clear inhibition of exosome release by tumour cells. Human tumours obtained from xenografts pretreated with PPI contained more CisPt as compared to tumours from xenografts treated with CisPt alone. Further analysis showed that in vivo PPI treatment induced a clear reduction in the plasmatic levels of tumour-derived exosomes which also contained lower level of CisPt. Altogether, these findings point to the identification of a double mechanism that human malignant melanoma use in resisting to a dreadful cellular poison such as cisplatin. This framework of resistance includes both low pH-dependent extracellular sequestration and an exosome-mediated elimination. Both mechanisms are markedly impaired by proton pump inhibition, leading to an increased CisPt-dependent cytotoxicity.

Human biomonitoring for Cd, Hg and Pb in blood of inhabitants of the Sacco Valley (Italy).

INTRODUCTION: The Sacco Valley (Lazio, Italy) is characterized by high density population and several industrial chemical productions that during the time had led to a substantial amount of by-products. The result was a severe environmental pollution of the area and in particular of the river Sacco. In 1991, the analysis of water and soils samples of three industrial landfills revealed the presence of organochlorine compounds and heavy metals. A research project named "Health of residents living in Sacco Valley area", coordinated by the regional Department of Epidemiology, was undertaken and financed to evaluate the state of health of the population living near those polluted areas. MATERIALS AND METHODS: Cd, Hg and Pb were quantified in 246 blood samples of potentially exposed residents of the Sacco Valley by quadrupole inductively coupled plasma mass spectrometry (Q-ICP-MS). RESULTS: Individuals who agreed to be sampled did not exhibit high levels of the elements. The distance from the river does not seem to be directly connected with the elements levels in blood. The contribution of these contaminants to the total intake due to ingestion of food was difficult to evaluate. The unclear trend of data would require a characterization of the polluted site with environmental sampling of different matrices.

[Epidemiologic studies on the health status of the population living in the Sacco River Valley]. / Indagini epidemiologiche sullo stato di salute della popolazione residente nell'area della Valle del Sacco.

UNLABELLED: OBIETTIVO: to analyze the health status of the population living in an area close to the Colleferro industrial plant. SETTING AND PARTICIPANTS: the area of the Sacco River Valley, Central Italy nearby Rome, has been heavily polluted over the years by industrial wastes deriving from the chemical industrial plant in Colleferro. In 2006, it was discovered that the herds of livestock were contaminated by beta-hexachlorocycloexane (ß-HCH, an industrial waste belonging, as well as lindane, to the group of hexachlorocycloexane isomers). MAIN OUTCOME MEASURES: the analyses of mortality and morbidity were carried out for the 1998-2007 period (calculation of standardized mortality ratios, SMR), and for the period 2003-2007 (calculation of standardized hospitalization ratios, SHR), respectively. The general population in the Lazio Region has been considered as reference. In addition, a biomonitoring study was conducted on a sample of the population living in 4 areas of the Sacco River Valley with different levels of exposure and the following persistent organic pollutants were measured in the blood (α, ß and γ-HCH, HCB p,p'-DDT and p,p'- DDE, 6 NDL-PCB congeners and 12 DL-PCBs, PCDDs and PCDFs), and heavy metals (Cd, Hg, Pb). RESULTS: cancer mortality in men was increased in the area (SMR=1.20), especially for specific cancer sites (stomach, larynx, lungs, pleura, myeloma); in women an excess of mortality from diabetes was detected (SMR=1.44). The analysis of morbidity indicated an excess of hospitalization for various cancers (larynx, myeloma) in men, for respiratory illness and asthma in both genders and for thyroid disease in women. The biomonitoring study found high mean concentration of ß-HCH (mean: 99.05 ng/g fat, SD=121.3), with higher levels in the population living along the river (mean=150 ng/g fat; SD=153.5), likely occurred through water and local food. CONCLUSION: the area of Colleferro has been polluted by multiple sources and the human population has been exposed to industrial chemicals, toxic substances in the workplace, and to the cumulative accumulation of organic pesticides especially through water and food.

The Italian Helpdesk under the Regulation (EC) No. 1272/2008 (CLP): three-year activity and experience (2009-2011).

INTRODUCTION: The Regulation (EC) No. 1272/2008 on classification, labelling and packaging of substances and mixtures (CLP) sets further obligations for manufacturers, importers, distributors, downstream users of substances either on their own or in mixtures. According to the European mandate, each Member State has constituted its National Helpdesk to provide advice to the interested parties on their duties under this Regulation. In Italy, the contact point for questions has been established at the National Centre for Chemical Substances of the Istituto Superiore di Sanità. FUNCTIONS: The responders of the Italian CLP Helpdesk process the requests that have been submitted by the dedicated website. Applicants are asked to complete the form with all the required information. The Helpdesk staff also take part in the European network of CLP, REACH and ECHA Helpdesks together with the European Commission and other parties, that is the HelpNet. RESULTS: The present paper describes the results of the three-year activity of the Italian CLP Helpdesk (2009-2011).

[Exploratory biomonitoring study among workers of livestock farms of the Taranto Province]. / Studio esplorativo di biomonitoraggio tra gli allevatori delle masserie della Provincia di Taranto.

OBJECTIVE: to conduct a survey on blood levels of metals and organochlorine compounds (dioxins and PCBs) among workers of livestock farms, and to identify the factors able to explain the magnitude and variability of the measured concentrations. DESIGN: exploratory human biomonitoring study. SETTING AND PARTICIPANTS: a sample of 45 workers from farms (masserie) located in the Province of Taranto was included in the study. The farms, located between 6 and 43 km from the industrial area, are family owned and animals (mainly sheep and goats) are locally reared outdoors. The study subjects have similar eating habits, including food consumption resulting from their production. RESULTS: manganese concentrations and, to a lesser extent, arsenic, cadmium and lead are in the medium-high range of levels observed in the Italian general population. This study does not identify factors able to explain the variability in the blood levels of metals. The amount of dioxins and PCBs, on the other hand, is strongly associated both with subjects age and the distance of farm from the industrial site. CONCLUSION: the results of this survey are suggestive of research hypotheses that need to be confirmed by biomonitoring studies of adequate design and size, in particular the hypothesis that the blood level of dioxins and PCBs decreases with increasing distance from the industrial site. We recommend ad hoc studies to better characterize the exposure of farmers (also associated with agricultural activities), and of the inhabitants of Taranto, to environmental persistent pollutants, also in terms of food matrices locally produced and consumed.

Dynamic reaction cell ICP-MS for determination of total As, Cr, Se and V in complex matrices: still a challenge? A review.

Mass interferences, caused by atomic or polyatomic species and having the same mass/charge ratio of the analyte, can be a severe limit for a reliable assay of trace and ultratrace elements by ICP-MS. The DRC™ technology uses a reaction gas to overcome these interferences. Reactions of charge exchange, atom transfer, adduct formation, condensation and analyte association/condensation are the main mechanisms. Interfering ions tend to react with the gas exothermally, while, the analyte reacts endothermally. Selecting the most appropriate reaction gas in DRC-ICP-MS is the very critical point for the determination of strongly interfered elements. A careful evaluation of the reaction mechanisms and the chemistry involved are required. The DRC allows the use of different gases, among them, ammonia (NH(3)), methane (CH(4)), hydrogen (H(2)) and oxygen (O(2)) are the most known, but there are other potentially useful gases like nitrous oxide (N(2)O), nitrogen oxide (NO), carbon dioxide (CO(2)), fluoromethane (CH(3)F), sulphur hexafluoride (SF(6)) and carbon disulfide (CS(2)). This paper provides a review on the analytical challenges for a reliable assay of As, Cr, Se and V by DRC-ICP-MS and illustrates different approaches and mechanisms involved in the analysis of polymers, biological fluids (serum, urine and whole blood), rock, soil and particulate matter.

Method validation for determination of cadmium and lead in offal by means of quadrupole inductively coupled plasma mass spectrometry.

Offal includes viscera and internal organs that have been removed from butchered animals, that may be either directly eaten or processed for the production of other foodstuff. Such organs are able to accumulate high concentrations of potentially toxic heavy metals posing a risk for human health when ingested. Because high levels of Cd and Pb may produce damages to humans, Commission Regulation no. 1881/2006 and its amendment established maximum levels for those two elements in edible bovine, porcine, and ovine offal. In the present study, a method based on microwave acid-assisted digestion and quadrupole inductively coupled plasma mass spectrometric quantification of Cd and Pb in offal was validated according to the EU common standards. The main parameters evaluated in the validation process were: the detection and the quantification limits (LoD, LoQ), the recovery, the repeatability, the within-laboratory reproducibility, the linearity range, and the standard measurement uncertainty. The results obtained for LoD and LoQ in microg kg(-1) were, respectively, Cd, 1.8 and 5.4; Pb, 5.1 and 15.5; meanwhile, the mean recovery was about 98% for Cd and 103% for Pb. Repeatability was around 5% for Cd and 4% for Pb. The expanded standard measurement uncertainty, expressed as percentage and with a coverage factor of 2, was estimated as follows: Cd, 4.9%; Pb, 8.7%. For both elements, the main contribution was due to the within-laboratory reproducibility of the measurements.

The network of the Italian laboratories: a proficiency test on the quantification of trace elements in serum.

A proficiency test on the quantification of Al, Cu, Mn, Se and Zn in serum was carried out to verify the performance of about 30 regional laboratories of the network of Italian laboratories. The exercise consisted of four runs in which the laboratories were free in choosing analytical methods to determine trace elements in freeze-dried animal serum. Laboratories performances were evaluated by the study of statistical functions as Coefficients of Variation (CV), Youden plot and Z-score value. As for Al, the results were generally characterized by good accuracy and precision, in spite of the low levels of the element (5-7 microg l(-1)). Copper determination had some problems only at low concentration (about 160 microg l(-1)--first run), in which an elevated number of anomalous data were found. Better data were achieved for Zn, for which anomalous values were mainly stragglers than outliers. Due to the low number of data for Mn (concentrations from 0.6 to 60 microg l(-1)) and Se (concentration from 45 to 106 microg l(-1)), a restricted statistical treatment was applied; for these elements high CV values were found (range from 30 to 80%). The results of this trial confirmed that participation in a proficiency test represents a benefit for all analytical laboratories. In fact, with few exceptions, most of the participants improved their performances in terms of Z-score values.