Evidence for persistent organochlorine pollutants in the human adrenal cortex.

Environmental pollutants may act as endocrine disruptors in animals. Organochlorine pesticides (OCPs) and polychlorinated biphenyls (PCBs) enter the food chain and may accumulate in the fatty animal tissues, including adrenals. To our knowledge, no previous study has investigated their presence in the human normal adrenal (NA) cortex and aldosterone-producing adenomas (APA). Surgical fragments of APA from 11 patients and NA from 8 kidney donors were analyzed for 16 PCBs congeners and 10 OCPs. A Matrix Solid-Phase Dispersion (MSPD) method for simultaneous determination of the target compounds in cortex homogenates was developed. A gas-chromatography triple quadrupole mass spectrometry (Triple Quad GC-MS) system was used for the analysis. Data were analyzed using Random Forest and Wilcoxon's rank-sum test. OCPs and PCBs were found in specimens from both types. A subset of pollutants characterized APA more than NA. Higher concentrations (µg g-1 ) in APA were observed for α-, ß-, and γ- Hexachlorocyclohexane (HCH) (1.48 ± 3.32 vs. 0.17 ± 0.19, P = 0.028; 2.81 ± 2.10 vs. 0.96 ± 0.98, P = 0.011; 2.16 ± 4.85 vs. 0.17 ± 0.26, P = 0.004, respectively), as well as for Hexachlorobenzene (HCB) and for PCBs 28, 52 and 101 (3.41 ± 3.11 vs. 0.97 ± 1.06, P = 0.021; 2.34 ± 4.68 vs. 0.25 ± 0.22, P = 0.039; 0.58 ± 1.19 vs. 0.06 ± 0.02, P = 0.002; 0.26 ± 0.43 vs. 0.05 ± 0.00, P = 0.001, respectively). Environmental organochlorine pollutants were shown to be present in the human normal and abnormal adrenal cortex, deserving future investigation on their possible role as adrenal endocrine disruptors in human disease. Copyright © 2017 John Wiley & Sons, Ltd.

Serum concentrations of persistent organic pollutants (POPs) in the inhabitants of a Sicilian city.

BACKGROUND: Polychlorinated biphenyls (PCBs) and organochlorine pesticides are common environmental contaminants that have been associated with human health problems. OBJECTIVES: To assess serum concentrations of several organochlorine contaminants in general population living in a city with an ancient agricultural tradition and to identify possible exposure sources in Sicily. METHODS: A cross-sectional study was conducted on 101 individuals. Each participant answered a face-to-face questionnaire submitted by well-trained personnel and provided a serum sample which was analyzed for the concentrations of PCBs, HCB, HCHs and DDTs by using gas-chromatography coupled with mass spectrometry. RESULTS: HCB, p,p'-DDE, PCB 138, PCB 153 and PCB 180 were detected in more than 80% of the study participants. The ng g(-1) lipid median concentrations were: 18.6 for HCB; 175.1 for p,p'-DDE; 22. for PCB 138; 32.5 for PCB 153 and 23.0 for PCB 180. PCB 153 and PCB 138, PCB 138 and PCB 180, PCB 153 and PCB 180, and p,p'-DDE and HCB showed a high correlation each other (p<0.05). HCB and p,p'-DDE concentrations were significantly higher in subjects >49 years old (adj-p=0.03 in 50-69 years old and adj-p<0.001 in >69 years old, respectively) whereas PCB 138, PCB 153 and PCB 180 concentrations were higher in males (adj-p=0.03), in subjects >69 years old (adj-p=0.04) and in current smokers (adj-p=0.04). CONCLUSIONS: The present study shows that serum concentrations of organochlorine compounds detected in subjects living in a small Sicilian city with ancient agricultural tradition are similar to those found in subjects living in urban areas of other countries. However, further investigations are needed to compare data from rural/urban areas in the same country, assessing correlations between serum concentrations of several chemical compounds and potential health effects in general population.

Farmers' exposure to herbicides in North Italy: assessment under real-life conditions in small-size rice and corn farms.

Although rice and corn are two main cash crops in Lombardy (North Italy) and their cultivation involves several thousands of farmers, risk assessment of pesticide exposure is rarely done, especially in small and medium sized enterprises. With the use of pads for environmental monitoring (OECD protocol), of pre- and post-exposure 24h urine collection for biological monitoring and of hand wash for hands' exposure, we measured the exposure of 28 agricultural workers to propanil and terbuthylazine in real-life working conditions. In propanil applicators, median daily exposure on the clothes was 73.5µmol per worker, while the exposure on the skin was 22.4µmol. For terbuthylazine, these exposures were 37.2µmol and 0.86µmol per worker, respectively. Median excretion of the propanil metabolite (3,4-dichloroaniline) after exposure was 84nmol in 24h urine, and 13nmol for the metabolite of terbuthylazine. Risk assessment performed by comparing to the AOELs of the applied active ingredients with an estimated internal dose, obtained based on the measured levels of skin and hand exposure and the percentage of dermal absorption of the active ingredients considered showed that 4 propanil workers, and no terbuthylazine workers, were overexposed. Our study helps define and confirm relationships between different exposure determinants, which can be used in the development of tools for risk assessment of exposure to pesticides in small and medium sized enterprises.

Integration of biological monitoring, environmental monitoring and computational modelling into the interpretation of pesticide exposure data: introduction to a proposed approach.

Open field, variability of climatic and working conditions, and the use of complex mixtures of pesticides makes biological and environmental monitoring in agriculture, and therefore risk assessment and management, very complicated. A need of pointing out alternative risk assessment approaches, not necessarily based on measures, but simple, user-friendly and reliable, feasible also in the less advanced situations and in particular in small size enterprises, arises. This aim can be reached through a combination of environmental monitoring, biological monitoring and computational modelling. We have used this combination of methods for the creation of "exposure and risk profiles" to be applied in specific exposure scenarios, and we have tested this approach on a sample of Italian rice and maize herbicide applicators. We have given specific "toxicity scores" to the different products used and we have identified, for each of the major working phases, that is mixing and loading, spraying, maintenance and cleaning of equipment, the main variables affecting exposure and inserted them into a simple algorithm, able to produce "exposure indices". Based on the combination of toxicity indices and exposure indices it is possible to obtain semiquantitative estimates of the risk levels experienced by the workers in the exposure scenarios considered. Results of operator exposure data collected under real-life conditions can be used to validate and refine the algorithms; moreover, the AOEL derived from pre-marketing studies can be combined to estimate tentative biological exposure limits for pesticides, useful to perform individual risk assessment based on technical surveys and on simple biological monitoring. A proof of principle example of this approach is the subject of this article.

Pesticide exposure and serum organochlorine residuals among testicular cancer patients and healthy controls.

The incidence of testicular cancer (TC) has been increasing worldwide during the last decades. The reasons of the increase remains unknown, but recent findings suggest that organochlorine pesticides (OPs) could influence the development of TC. A hospital-based case-control study of 50 cases and 48 controls was conducted to determine whether environmental exposure to OPs is associated with the risk of TC, and by measuring serum concentrations of OPs, including p,p'-dichlorodiphenyldichloroethylene (p,p'-DDE) isomer and hexachlorobenzene (HCB) in participants. A significant association was observed between TC and household insecticide use (odds ratio [OR] = 3.01, 95 % CI: 1.11-8.14; OR(adjusted) = 3.23, 95 % CI: 1.15-9.11). Crude and adjusted ORs for TC were also significantly associated with higher serum concentrations of total OPs (OR = 3.15, 95 % CI: 1.00-9.91; OR(adjusted) = 3.34, 95 % CI: 1.09-10.17) in cases compared with controls. These findings give additional support to the results of previous research that suggest that some environmental exposures to OPs may be implicated in the pathogenesis of TC.

Occupational exposure to antineoplastic drugs in seven Italian hospitals: the effect of quality assurance and adherence to guidelines.

In health care facilities, dermal contact and inhalation are considered to be the main routes of exposure to cytotoxic antineoplastic drugs (ADs). Hand-to-mouth contamination or accidental needle sticks as well as events due to inadequate disposal may also contribute to exposure. In order to measure the extent of contamination, biological and environmental monitoring are essential tools for routine testing. Moreover, reliable sampling and analytical procedures are required. During the last decade, several methods have been developed and validated. The appropriate analytical techniques were used to quantify even very low levels of some of the more commonly used ADs, such as cyclophosphamide, 5-fluoruracil, taxol, anthracyclines, and platinum-compounds. The main objective of this study is to assess the adherence to existing standards of practice through an effective monitoring program, including environmental and biological measurements. In seven hospitals located in Northern-Central Italy, periodic surveys were scheduled to verify continuing compliance with guidelines over a 5-year period. All biological samples were found to be below detection limits and a progressive, significant decrease in workplace contamination was observed. Our results confirm that a cost-effective monitoring regime, including fast and simple sample pre-treatment procedures, simultaneous determination of the analytes and their metabolites, validated procedures including uncertainty evaluation, and periodic surveys, is the adequate approach for the collection of reliable exposure data and hence for effective intervention.

A simple and fast method for the determination of selected organohalogenated compounds in serum samples from the general population.

Polychlorinated biphenyls (PCBs) and organochlorine pesticides (OCPs) have been widely used in industry and agriculture. Due to their persistence and bioaccumulation, they were globally spread in the environment and may still be found in environmental and biological media, despite the international restrictions on production and use. The main aim of our study was to develop a simple and fast method suitable for the establishment of the reference values for 15 PCB congeners and 16 OCPs in general population subgroups. A cost- and time-saving screening procedure using gas chromatography coupled with low-resolution mass spectrometry, was improved and validated before application to the analysis of real samples. The overall method was validated including uncertainty measurement. Preliminary field data were collected from 95 volunteers living in two Italian areas. HCB, p,p'-DDE, PCB 153, PCB 138 and PCB 180 were the most frequently detected compounds. Age and residence area were found to be significant variables for the most abundant compounds, while no correlation between serum concentrations and gender was observed. Our results suggest that long-banned substances, including PCBs and the pesticides HCB and DDT's breakdown product, are still detectable in the general population.

Immunomodulatory effects of the herbicide propanil on cytokine production in humans: In vivo and in vitro exposure.

Propanil, 3,4-dichloropropionanilide, a commonly used herbicide, has been shown to induce effects on the mouse immune system. The aim of this study was to assess the immunotoxicity of propanil in occupationally exposed agricultural workers and to characterize its molecular mechanism of action. Seven agricultural workers intermittently exposed to propanil and 7 healthy matched controls entered the study. Data were collected through physical examination, and laboratory investigations addressed at the main serum, cellular, and functional immune parameters. The levels of exposure were assessed by determining the urine concentration of the major propanil metabolite, 3,4-dichloroaniline. The investigation of serum, cellular, and functional immune parameters suggested that propanil exposure results in a modest immunomodulatory effect, characterized by an increase in the plasma level of IgG(1) and in LPS-induced IL-6 release and, by a reduction in PHA-induced IL-10 and IFN release, associated with a reduced IFN/IL-4 ratio. As observed, following in vivo exposure, in vitro treatment of human peripheral blood leukocytes with propanil resulted in a dose-dependent reduction in PHA-induced IFN-gamma and IL-10 production, while LPS-induced TNF-alpha production was not affected indicating a direct effect of propanil on selected immune parameters. We demonstrated that propanil interfering with PHA-induced intracellular calcium increase modulated IL-10 and IFN-gamma transcription and translation, which indicates that propanil acts on early events triggered by PHA. Overall, our results suggest that human exposure to propanil has slight immunomodulatory effects, and point out that the inhibition of the PHA-induced intracellular calcium rise is an important target of propanil. These findings improve our understanding of the mechanism underlying propanil-induced immunotoxicity.

Simultaneous determination of gemcitabine, taxol, cyclophosphamide and ifosfamide in wipe samples by high-performance liquid chromatography/tandem mass spectrometry: protocol of validation and uncertainty of measurement.

Measurable levels of anticancer agents are still detected on work surfaces in health-care settings. However, application of recent guidelines for the protection of workers' safety and health has resulted in lowered contamination levels. To assess occupational exposure to antineoplastic agents, very sensitive and specific procedures for environmental sampling and analysis are therefore needed. In the present study an assay for simultaneous determination of gemcitabine, taxol, cyclophosphamide, and ifosfamide in wipe samples, using two internal standards (trofosfamide and cephalomannine), was developed and validated by high-performance liquid chromatography/tandem mass spectrometry (HPLC/MS/MS). Solid-phase extraction (SPE) was used for sample concentration and cleanup. The assay was found to be linear up to 1000 ng/wipe, with limits of quantitation of 25.0 ng/wipe for gemcitabine and taxol, and 12.5 ng/wipe for cyclophosphamide and ifosfamide. In order to investigate the effectiveness of the surface sampling, removal efficiency tests were repeated on different types of surfaces. Recovery rates of between 62 and 81% were obtained at two contamination levels (50.0 and 250 ng/100 cm2). Precision and trueness were determined on three different days. The within-day precision was found to be always less than 12.1% for all the analytes. The overall precision, expressed as relative standard deviation (RSD), was always less than 9.4%. Recoveries varying from 75.0 (gemcitabine) to 95.0% (taxol) were obtained at three levels. In order to obtain a quantitative indication of the quality of the result, the overall uncertainty of measurement (UOM) was evaluated according to the EURACHEM/CITAC guide. The relative combined uncertainty was found to be always less than 9.5%. The relative expanded uncertainty was also calculated, at three contamination levels.

Residual hazard assessment related to handling of antineoplastic drugs: safety system evolution and quality assurance of analytical measurement.

Despite improvement of operating procedures and publication of safety guidelines, contamination is still observed in healthcare settings where antineoplastic drugs (ADs) are handled. Even after cleaning work areas, some residual contamination may still be present. Zero percent contamination is not a realistic goal, but the scientific community should set zero contamination as its main goal. The strategies to reach this objective may be traced based on the followings: (a) a wider number of drugs should be monitored; (b) safety equipment and devices must be available to the workers; (c) the likely source of widespread contamination in workplaces is the safety cabinet; (d) direct determination of the parent drug or its metabolite in urine is the recommended approach because it provides higher sensitivity and specificity; (e) reliable analytical methods are necessary to measure the extent of contamination; and (f) analytical methods intended to be applied for routine testing must be assessed through method validation studies. These studies rely on the determination of overall method performance parameters including uncertainty measurement. Our laboratory has developed and validated a number of analytical methods for the determination of several drugs in environmental and biological samples. Surveys were carried out in several hospitals, and there has been progressive, significant decrease in the number of positive samples, mostly due to the improvement of working procedures and safety measures.

[Polychlorinated biphenyl serum levels in two Italian population groups: sex pattern and correlation with age]. / Livelli serici di policlorobifenili in due gruppi di popolazione Italiana: distribuzione dei congeneri e correlazione con l'età.

OBJECTIVE: To examine the congener-specific concentrations and patterns of polychlorinated biphenyls in the Italian general population. DESIGN: Two population groups were selected and monitored for the determination of up to 60 congeners in human serum. A very detailed questionnaire was administered to each participant to collect information concerning demographic, lifestyle, and medical factors. Statistical analyses were performed to examine the relationships between PCB serum levels and age. SETTING: Northern and Central Italy. PARTICIPANTS: 162 subjects from Novafeltria, Central Italy and 164 subjects from Pavia, Northern Italy. MAIN OUTCOME MEASURE: Serum concentrations of both coplanar and non-coplanar PCB congeners. Relationship between PCB levels and age. RESULTS: The mean concentration of total PCBs was found to be 2.48 and 3.93 microg/l for Novafeltria and Pavia, respectively. As waited, the most abundant congeners were CB-153, CB-138, CB-180, and CB-170. Age resulted to be the most significant determinant of PCB levels. Slight differences in the congener profile were observed in the two population groups. CONCLUSIONS: As expected, a strong correlation between age and PCB levels has been observed. It is remarkable that the correlation increased with increasing chlorination in both the population groups. Associations with other variables, such as gender, food and alcohol consumption, and cigarette smoking, appeared to be inconsistent.

Determination of dichloroanilines in human urine by gas chromatography/mass spectrometry: validation protocol and establishment of Reference Values in a population group living in central Italy.

3,4- and 3,5-Dichloroanilines (DCAs) are common markers of some non-persistent pesticides, e.g. linuron, diuron, vinclozolin, and iprodione. The general population may be exposed to these DCAs and/or their precursors mainly through diet. Since adverse effects on human health, such as endocrine disruption, have been reported, biological monitoring is essential for exposure assessment both of occupationally exposed subjects and of the general population. A highly sensitive and selective gas chromatography/mass spectrometry (GC/MS) method has been developed for the determination of 3,4- and 3,5-DCAs in urine using 4-chloro-2-methylaniline as an internal standard. The selected ion monitoring (SIM) mode was employed for quantitation of the analytes. The sample treatment procedure is simple and fast and no derivatization is required. The overall method was validated including uncertainty measurement. The limit of detection (LOD) and the lower limit of quantitation (LLOQ) were determined to be 0.005 and 0.010 microg/L for both analytes. The method was then applied to the establishment of reference values for a population group living in a rural area of central Italy (Novafeltria, Marche). A total of 151 out of 153 samples were found to be positive for 3,5-DCA, and 81.7% were positive for 3,4-DCA. For this group, 3,4-DCA levels ranged from 0.01 to 6.19 microg/L, while 3,5-DCA urinary concentrations were between 0.02 and 6.71 microg/L.

Reference values of coplanar and non-coplanar PCBs in serum samples from two Italian population groups.

The main goal of this study is to establish the reference values of individual Polychlorinated biphenyl (PCB) congeners in non-occupationally exposed subjects. Since the PCB pattern in human serum is related to the living area, two different population groups from North and Central Italy, were compared. Serum concentrations of both coplanar and non-coplanar PCB congeners were measured by using gas chromatography coupled with low-resolution mass spectrometry (HRGC-LRMS). A fast and reliable method for the determination of 60 congeners had been previously validated. Its reliability was further verified by using high-resolution mass spectrometry. Thirty-one congeners out of 60 were found at detectable concentrations in at least one sample. The mean value for total PCBs was found to be 2.48 and 3.93 microg/L for the two population groups. Eight dioxin-like PCBs were detected. In accordance with the findings from the literature, the most abundant congeners were found to be 153, 138, 180, and 170. Both univariate and multivariate analysis showed that age is a significant determinant of PCB concentrations. The correlation increased with increasing chlorination. Slight differences in the PCB pattern were observed in the two population groups.

Validation protocol and analytical quality in biological monitoring of occupational exposure to antineoplastic drugs.

Occupational exposure to antineoplastic cytostatic drugs has been recognized as a potential health hazard since the seventies. Safety guidelines and recommendations have been published in several countries in order to improve operating procedures and keep exposure levels as low as possible. Nevertheless, contamination still occurs. With a view to preventing exposure, standardized authorized methods and a mandatory monitoring system appear to be an essential prerequisite. First of all, reliable analytical methods are necessary, and a wide number of drugs should be monitored, on the basis of the continual change in the treatment protocols. In fact, most of the methods available in the literature were not properly validated, and a lack of details concerning quality assurance is clearly observed. When assessing analytical methods applied for routine testing, validation studies are of utmost importance. Not only performance parameters, such as sensitivity, specificity, LOD and LLQ, should therefore be determined, but also the uncertainty of measurement, which gives a measure of the confidence that can be placed on the results.

Critical evaluation of a high-throughput analytical method for polychlorinated biphenyls in human serum: which detector for the establishment of the reference values?

A rapid screening method, previously developed and validated for the determination of 60 PCB congeners in human serum by using high-resolution gas chromatography coupled with low-resolution mass spectrometry, was applied to the establishment of the reference values in the general population. The reliability of this method was further confirmed by analyzing ten serum samples, obtained from the same population group involved in the original study, by electron impact ionization with either low- or high-resolution mass spectrometry. Analyses in electron capture negative ionization mode were also performed for the determination of 'dioxin-like' isomers. The data were found to be in good accordance with one another, especially for the congeners of highest environmental significance. Then, 162 real serum samples were analyzed for the presence of PCBs. Comparison with the results from other studies reported in the literature showed that a similar profile was observed; the most abundant congeners were #153, #180, #138 and #170. A statistical analysis was performed to determine whether there were significant correlations between PCB concentrations and specific variables such as age, gender, and dietary habits.

Determination of coplanar and non-coplanar polychlorinated biphenyls in human serum by gas chromatography with mass spectrometric detection: electron impact or electron-capture negative ionization?

A time- and cost-saving method for the congener-specific analysis of polychlorinated biphenyls (PCBs) in human serum has been developed and validated. After two fast extraction and clean-up steps, analyses were performed using gas chromatography coupled with mass spectrometry with single ion monitoring (GC/SIM-MS), either in electron impact (EI) or electron-capture negative ionization (ECNI) mode. For the determination of dioxin-like congeners, an improvement in EI-MS sensitivity is desirable and use of NI is thus preferred. The procedure was validated for 12 dioxin-like congeners by analyzing spiked samples on three different days and using (13)C(12)-labelled analogues as internal standards. When using an NCI source, the limit of quantification was assessed at 0.01 microg/L, except for PCBs #77 and #81, which cannot be reliably detected below 0.05 microg/L. For the lower chlorinated non-dioxin-like congeners, NI offers less selectivity because of limited fragmentation. Electron impact ionization and electron-capture negative ionization mode can therefore be considered to be complementary for the determination of PCB congeners in the general population.

Biological and environmental monitoring of hospital personnel exposed to antineoplastic agents: a review of analytical methods.

In order to assess occupational exposure of hospital personnel involved in the preparation and administration of antineoplastic drugs, biological and environmental monitoring are essential to identify the main exposure routes and to quantify potential health risks. If workplace contamination cannot be completely avoided, it is of utmost importance to reduce exposure to the lowest possible levels. To this aim, not only do education and training of the exposed subjects play an important role, but accurate standardized sampling techniques and analytical methods are also required. A critical overview of the most significant methods available in the literature is presented and their value is discussed, especially with respect to their sensitivity and specificity. In addition, attention is given to validation procedures and, consequently, to their reliability. The results from the most important surveys carried out at hospital departments are also discussed, with a view to improving both monitoring strategies and moreover working conditions.

A rapid screening method for routine congener-specific analysis of polychlorinated biphenyls in human serum by high-resolution gas chromatography with mass spectrometric detection.

A screening method using gas chromatography/mass spectrometry in the single ion monitoring (SIM) mode for the routine congener-specific analysis of polychlorinated biphenyls (PCBs) in human serum has been developed and validated. A fast procedure incorporating both liquid-liquid and solid-phase extraction was used for clean up and enrichment. Interday and intraday precision and accuracy were assessed for the 12 congeners having dioxin-like activity and PCBs #180 and #170 by analyzing spiked samples on three subsequent days and using (13)C(12)-labelled analogues as internal standards. Furthermore, the relative errors for six non-dioxin-like congeners (PCBs #28, #52, #101, #138, #153 and #180) were measured by using certified reference materials, providing good accuracy at two different concentration levels. The limits of quantification and of detection were 0.5 and 0.2 micro g/L in serum for each congener, respectively. The method allows the chromatographic separation of at least 35 congeners and can therefore be applied to the routine monitoring of the general population for both dioxin-like and non-dioxin-like PCB congeners.