Natalia Quinete

Highly selective and automated online SPE LC–MS3 method for determination of cortisol and cortisone in human hair as biomarker for stress related diseases

Hair analysis has been increasingly used to establish long-term biomarkers of exposure to both endogenous and exogenous substances, with a special emphasis on steroidal hormones. Hair cortisol and cortisone have been associated to physiological and psychological strains, anxiety and depression. Hair is a very complex matrix, which might jeopardize analyte detection at low concentrations. A new, highly selective and sensitive method based on fragments of second order, MS(3) (MS/MS/MS), was developed and validated for the analysis of hair cortisol and cortisone. An online solid phase extraction was performed on a C8 restricted access material (RAM) phase following by separation on a reversed-phase C18 column using methanol and 0.02% ammonium hydroxide as mobile phase. The developed method required minimal sample preparation and the injection of only 50 µL of sample leading to a LOQ of 2 pg mg(-1). Good linear responses were observed in the range 2-200 pg mg(-1) (R(2)>0.99) and extraction recoveries ranged between 77-125% and 70-123% for cortisol and cortisone, respectively. Intra- and inter-assay coefficients of variation were between 1.4 and 14%. In order to evaluate the applicability of the method, preliminary tests (N=33) were conducted in 3 cm hair samples (close to scalp) of healthy volunteers with an age range of 4-63. Average concentrations in hair were 12.7±14 pg mg(-1) and 41.6±42 pg mg(-1) for cortisol and cortisone, respectively. Further investigations on cortisol and cortisone as biomarkers for chronic psychological strain will be assessed as a next step.

Analytical approaches for the determination of PCB metabolites in blood: a review

Polychlorinated biphenyls (PCBs) are among the most ubiquitous pollutants in the environment, and their metabolism leads to the formation of hydroxylated PCBs (OH-PCBs) and methyl sulfone PCBs (MeSO2-PCBs). These metabolites are generally more hydrophilic than the parent compound, and therefore are more easily eliminated from the body. However, some congeners have been shown to be strongly retained in human blood, binding to transthyretin with an affinity that is, in general, greater than that of the natural ligand thyroxin itself, which could result in toxicological effects, particularly on the thyroid system. Currently available analytical methods require, in general, extensive sample preparation, which includes a series of time-consuming and low-throughput liquid–liquid and back extractions, evaporations, several cleanup steps, and in some cases, derivatization prior to analysis by gas chromatography (GC) or liquid chromatography (LC) coupled with mass spectrometry (MS). Recent developments in the use of LC coupled with tandem MS (MS/MS) have brought some improvements in terms of sample preparation for the determination of PCB metabolites in blood, although there are still possibilities for continued development. The selected literature has evidenced few studies of LC–MS/MS-based methods, a lack of analytical standards, nonassessment of lower-chlorinated OH-PCBs, and scarce attention to MeSO2-PCBs in blood. This review aims to evaluate critically the currently available analytical methods for determination of OH-PCBs and MeSO2-PCBs in blood.

Fast determination of hydroxylated polychlorinated biphenyls in human plasma by online solid phase extraction coupled to liquid chromatography-tandem mass spectrometry.

Hydroxylated polychlorinated biphenyls (OH-PCBs) have been shown to be strongly retained in human blood causing endocrine-related toxicity, particularly on the thyroid system. Traditionally, analytical methods for the determination of OH-PCBs require labor-intensive and long-time consuming sample preparation with several extraction, evaporation and cleanup procedures steps and, in some cases, derivatization prior to the analysis by gas or liquid chromatography-mass spectrometry (GC-MS or LC-MS). The present study developed and validated a novel, sensitive and high throughput online solid phase extraction (SPE) method coupled to LC-tandem mass spectrometry (MS/MS) for the separation and quantitation of relevant congeners of OH-PCBs in human plasma. The developed method presented limits of quantification (LOQ) ranging from 0.02 to 0.5 ng mL(-1) and extraction recoveries from 71 to 134% for all congeners, requiring small amount of sample (only 100 μL) and minimal sample preparation. In order to evaluate the applicability of the method, preliminary tests (N = 93) were conducted in plasma from individuals occupationally exposed to very high levels of PCBs in a German cohort. Penta-through hepta-chlorinated OH-PCBs were the predominant congeners in human plasma with concentrations up to 44.5 ng mL(-1), while lower chlorinated OH-PCBs were occasionally detected. In addition, a new PCB 28 metabolite has been synthesized and identified for the first time in human plasma and associations between OH-PCBs and their parent compounds in the studied cohort were also assessed.

PCB and PBDE levels in a highly threatened dolphin species from the Southeastern Brazilian coast.

In the Northern coast of Rio de Janeiro State is located the major urban centers of the oil and gas industry of Brazil. The intense urbanization in recent decades caused an increase in human use of the coastal areas, which is constantly impacted by agricultural, industrial and wastewater discharges. Franciscana dolphin (Pontoporia blainvillei) is a small cetacean that inhabits coastal regions down to a 30 m depth. This species is considered the most threatened cetacean in the Western South Atlantic Ocean. This study investigated the levels of 52 PCB congeners and 9 PBDE congeners in liver of nine individuals found stranded or accidentally caught between 2011 and 2012 in the Northern coast of Rio de Janeiro. PCB mean levels ranged from 208 to 5543 ng g(-1) lw and PBDEs mean concentrations varied between 13.84 and 36.94 ng g(-1) lw. Contamination patterns suggest the previous use of Aroclor 1254, 1260 and penta-BDE mixtures in Brazil. While still few studies have assessed the organic contamination in cetaceans from the Southern Hemisphere, including Brazil, the levels found in this study could represent a health risk to these endangered species.

Assessment of organochlorine pesticide residues in Atlantic Rain Forest fragments, Rio de Janeiro, Brazil.

A superficial water quality survey in a watershed of the Paraíba do Sul River, the main water supply for the most populated cities of southeastern Brazil, was held in order to assess the impact of the expansion of agricultural activity in the near border of the Atlantic Rain Forest. The aim of this study was to investigate the presence of priority organochlorine pollutants in soils and superficial waters of Atlantic rainforest fragments in Teresópolis, Rio de Janeiro State. Soil sample preparations were compared by using ultrasound, microwave assisted extraction and Soxhlet extraction. Recoveries of matrix spiked samples ranged from 70 to 130%. Analysis of a certified soil material showed recoveries ranging from 71 to 234%. Although low concentrations of organochlorine residues were found in water and soil samples, this area is of environmental importance and concern, thus demanding a monitoring program of its compartments.

PCB 28 metabolites elimination kinetics in human plasma on a real case scenario: Study of hydroxylated polychlorinated biphenyl (OH-PCB) metabolites of PCB 28 in a highly exposed German Cohort

Polychlorinated biphenyls (PCBs) are suspected of carcinogenic, neurotoxic and immunotoxic effects in animals and humans. Although background levels of PCBs have been slowly decreased after their ban, they are still among the most persistent and ubiquitous pollutants in the environment, remaining the subject of great concern. PCB 28 is a trichlorinated PCB found in high concentrations not only in human plasma but also in indoor air in Europe, yet little is known about its metabolic pathway and potential metabolites in humans. The present study aims to elucidate the kinetics of metabolite formation and elimination by analyzing four hydroxylated PCBs (OH-PCBs) in human plasma as potential metabolites of the PCB 28 congener. For this purpose, the study was conducted in plasma samples of highly PCB-exposed individuals (N=268), collected from 2010 to 2014 as a representation of a real case scenario with longitudinal data. OH-PCBs have been predicted, synthesized in the course of this study and further identified and quantitated in human plasma. This is the first time that previously unknown PCB 28 metabolites have been measured in human plasma and half-lives have been estimated for PCB metabolites, which could then provide further understanding in the toxicological consequences of exposure to PCBs in humans.

Determination of hydroxylated polychlorinated biphenyls (OH-PCBs) in human urine in a highly occupationally exposed German cohort: New prospects for urinary biomarkers of PCB exposure

The present study evaluates for the first time the determination of 20 hydroxylated polychlorinated biphenyl (OH-PCB) congeners and their glucuronide and sulfate conjugates in urine as a biomarker of exposure to PCBs in humans. Thereby, a fast, sensitive and selective online solid phase extraction (SPE) method coupled to LC-MS/MS was validated for the determination of OH-PCBs in human urine, being previously successfully developed and applied for the separation and quantitation of OH-PCBs in human plasma. The lowest limit of quantification (LLOQ) ranged from 0.01 to 0.19ngmL-1 and average extraction recoveries from 79 to 125% for all hydroxylated congeners. Within-run precision and between-run precision were between 2 and 17%. Extraction recovery tests were also performed in urine with different creatinine contents (0.52-3.92gL-1) for an estimation of matrix influences and ranged between 69 and 125%. In order to evaluate the applicability of the method, the study was conducted in three different groups, which were distinctly separated as non-exposed to known sources of PCBs (N=21), low-to-moderate PCB-exposed individuals (N=25) and highly occupationally PCB-exposed individuals (N=25), which included workers of a transformer recycling plant, their relatives and workers of surrounding companies from a German cohort. As part of the biomonitoring program HELPcB (Health Effects in High-Level Exposure to polychlorinated biphenyls), urine and blood samples were collected annually from 2010 to 2014. In this way, OH-PCB elimination profile in urine over time, correlations between OH-PCB levels in human plasma and urine, and associations with their parent compounds in plasma of the studied PCB cohort could be also assessed. Tri-chlorinated OH-PCBs were the predominant congeners in urine with concentrations up to 174ngmL-1. High chlorinated OH-PCBs (penta- through hepta-chlorinated OH-PCBs) were also frequently detected in urine samples from non-exposed and occupationally exposed individuals, although levels were in general very low or lower than LLOQ.

An assessment of PCB and PBDE contamination in two tropical dolphin species from the Southeastern Brazilian coast

PCBs and PBDEs were determined in two dolphin species, Sotalia guianensis and Steno bredanensis, from an upwelling system off the Central-northern coast of Rio de Janeiro, Brazil. PCB levels varied from 0.040 to 0.75 μg g(-1) lw in muscles and from 0.022 to 1.32 μg g(-1)lw in liver samples from S. guianensis. In S. bredanensis, values varied from 0.085 to 11.3 μg g(-1) lw in muscles and from 0.024 to 18.6 μg g(-1) lw in livers. PCB-138, -153 and -180 were the major PCB congeners detected in both species, while BDE-47 was the predominant PBDE congener found in both species. Higher concentrations in S. bredanensis were possibly related to the different feeding habits for both delphinid species. These results contribute to extend the database on organic contamination in cetaceans from the southern hemisphere, understanding their distribution and environmental fate in Southeastern Brazil.

Effects of psychosocial work characteristics on hair cortisol - findings from a post-trial study

Abstract Prolonged work stress, as indicated by the effort-reward imbalance (ERI) model, jeopardizes health. Cortisol represents a candidate mechanism connecting stress to ill health. However, previous findings appear inconclusive, and recommendations were made to assess work stress at multiple time points and also to investigate ERI (sub-)components. This study therefore examines the effects of two single time points, as well as the mean and change scores between time points of ERI and its components on hair cortisol concentration (HCC), a long-term cortisol measurement. Participants were 66 male factory workers (age: 40.68 ± 6.74 years; HCC: 9.00 ± 7.11 pg/mg), who were followed up after a stress management intervention (2006–2008). In 2008 (T1) and 2015 (T2), participants completed a 23-item ERI questionnaire, assessing effort, the three reward components (esteem, job security, job promotion) and over-commitment. In 2015, participants also provided a 3-cm hair segment close to the scalp for HCC analysis, as well as information on relevant confounders (i.e. medication intake, age, work characteristics, socioeconomic and lifestyle factors, number of stressful life events). Linear regressions revealed hardly any cross-sectional or longitudinal effect of ERI and its components on HCC. Only the change scores between T1 and T2 of job security were negatively associated with lower HCC in unadjusted (β = −.320; p = .009) and adjusted (β = −.288; p = .044) models. In this study, only a decrease of perceived job security over time was significantly associated with higher HCC, and other predictors were not related to this outcome. Especially after correction for multiple testing, this study revealed just a weak association of different psychosocial work measurements with HCC. Lay summary This study showed that an increase in perceived job insecurity is correlated with higher levels of the stress hormone cortisol. The higher levels of cortisol might represent a biological explanation for the negative health effects of job insecurity. The association was, however, relatively low, and more and more voices are questioning whether cortisol in hair is a reliable marker for perceived work stress.