Developmental toxicity and mechanism of dibutyl phthalate and alternative diisobutyl phthalate in the early life stages of zebrafish (Danio rerio).

Diisobutyl phthalate (DiBP), is widely chemical replacement for Dibutyl phthalate (DBP). Although DBP and DiBP have been detected in surface water worldwide, few studies to date have systematically assessed the risks of DBP and its alternatives to aquatic organisms. The present study compared DBP and DiBP for their individual and joint toxicity as well as thyroid hormone levels in zebrafish embryo. Transcripts of key genes related to the hypothalamic-pituitary-thyroid (HPT) axis were investigated in developing zebrafish larvae by application of real time polymerase chain reaction. The median half-lethal concentrations of DBP and DiBP to zebrafish at 96 h were 0.545 mg L-1 and 1.149 mg L-1, respectively. The joint toxic effect of DBP-DiBP (0.25-0.53 mg L-1) with the same ratio showed a synergistic effect. Thyroid hormones levels increased with exposure to 10 µg L-1 of DBP or 50 µg L-1 of DiBP, and exposure to both compounds significantly increased thyroid gland-specific transcription of thyroglobulin gene (tg), hyronine deiodinase (dio2), and transthyretin (ttr), indicating an adverse effect associated with the HPT axis. Molecular docking results indicated that DBP (-7.10 kcal/M and -7.53 kcal/M) and DiBP (-6.63 kcal/M and -7.42 kcal/M) had the same docking energy with thyroid hormone receptors. Our data facilities an understand of potential harmful effects of DBP and its alternative (DiBP).

Novel miniaturized passive sampling devices based on liquid phase microextraction equipped with cellulose-grafted membranes for the environmental monitoring of phthalic acid esters in natural waters.

Phthalic acid esters (PAEs) are considered endocrine disruptors and potential carcinogens. Consequently, efficient and accurate environmental monitoring of trace levels of these organic pollutants is necessary to protect the population against their hazardous effects. Passive sampling techniques have gained notoriety for environmental monitoring and have been proven highly sensitive to temporal variations. This study developed a miniaturized passive sampling device (MPSD) based on hollow fiber liquid-phase microextraction (HF-LPME). The devices were calibrated in the laboratory using an automated calibration system. The results demonstrated the first-order uptake ranges for Diethyl phthalate (DEP), Diisobutyl phthalate (DiBP), Dibutyl phthalate (DBP), Benzyl butyl phthalate (BBP) and Bis(2-ethylhexyl phthalate) (DEHP) between 30 min and 24 h with sampling rates equivalent to 0.009; 0.021; 0.033; 0.085 and 0.003 mL h-1 respectively (R2 between 0.88 and 0.99). The calibrated devices were deployed in 12 marginal lagoons, stretching approximately 330 km along the main river. The extracts recovered from the devices were analyzed by gas chromatography (GC), resulting in the identification and quantification of DEP (0.697-13.7 ng L-1), DiBP (0.100-4.43 ng L-1), DBP (0.014-1.21 ng L-1), BBP (0.218-5.67 ng L-1), and DEHP (0.002-2.24 ng L-1). Despite being frequently identified, DEHP concentrations were well below the maximum established limits, revealing a good water quality in terms of the target PAEs. In contrast, screening the extracts using GCxGC was possible to detect other hazardous pollutants such as pesticides, drugs, and their metabolites. The described device was effective and reliable, providing accurate PAE measurements following short exposure periods. In this sense, its deployment during emergency operations, such as accidental discharges of industrial effluents into natural waters, could continuously and cost-effectively monitor water quality.

Eucalyptol relieves the toxicity of diisobutyl phthalate in Ctenopharyngodon idellus kidney cells through Keap1/Nrf2/HO-1 pathway: Apoptosis-autophagy crosstalk and immunoregulation.

Diisobutyl phthalate (DiBP), one of the commonly used plasticizers in industry, is an endocrine disruptor and environmental contaminant that can persist in water and threaten the health of aquatic creatures. Eucalyptol (Euc), a monoterpenoid extracted from plants, has been proved to have anti-inflammatory, antioxidant, and detoxification properties. However, the protective mechanism of Euc against cell injury caused by DiBP exposure and the involvement of apoptosis, autophagy, and immunity remains unknown. In the current investigation, 27.8 µg/mL DiBP or/and 20 µM Euc has been applied to Ctenopharyngodon idellus kidney (CIK) cells for 24 h. The findings showed that exposure to DiBP raised intracellular ROS levels, inducing oxidative stress, and enhanced the rate of apoptosis as well as the expression of the apoptotic markers Bax, Caspase3, Caspase9, and Cytc while decreasing the expression of Bcl-2. Furthermore, DiBP inhibited IL-2, IFN-γ, Hepcidin-1, and ß-defensin expression and elevated TNF-α, and IL-1ß levels, causing immune dysfunction. DiBP and Euc co-treatment significantly activated the Keap1/Nrf2/HO-1 pathway, restored antioxidant enzyme activity, and elevated autophagy pathway-associated genes ATG5, Beclin1, and LC3B decreased p62 expression, enhanced cell autophagy, reduced apoptosis, and improved immunity. In conclusion, Euc promotes autophagy, alleviates DiBP-induced apoptosis, and improves immunological dysfunction in CIK cells by regulating the Keap1/Nrf2/HO-1 pathway. These results demonstrated the threat of DiBP exposure to fish while providing a theoretical foundation for using Euc in aquaculture.

Development of ecological risk assessment for Diisobutyl phthalate and di-n-octyl phthalate in surface water of China based on species sensitivity distribution model.

Phthalic acid esters (PAEs) are commonly used as plasticizer and are emerging concern worldwide for potent adverse effects of aquatic organisms. Certain PAEs were often detected in different environmental matrices but related toxicity data were still lacking to support their risk assessment. The study investigated the acute toxicity of Diisobutyl phthalate (DiBP) and Di-n-octyl phthalate (DnOP) using 6 Chinese resident aquatic organisms from 3 phyla and 6 species and constructed the species sensitivity distribution (SSD) models for ecological risk assessment. Lethal concentration 50% (LC50) ranges of DiBP and DnOP were 4.89-21.45 mg/L and 1.45-1200 mg/L, respectively. The derived acute and chronic predicted no-effect concentrations (PNECs) based on log-normal model of water were 0.54 and 0.04 mg/L for DiBP and 0.23 and 0.05 mg/L for DnOP, respectively. The ERA for DiBP and DnOP in the surface water and sediment of China was conducted. Water samples of Haihe Rive (RQ = 0.41) and Hun River (RQ = 0.16) of DiBP showed medium risk. And sediment samples of Yellow River (RQ = 0.71) and Chao Hu Lake (RQ = 0.42) of DiBP showed medium risk. Meanwhile, the above water and sediment samples (RQ＜0.1) of DnOP showed low risk.

Cumulative risk assessment of dietary exposure to phthalates in pregnant women in Beijing, China.

Diet is an important exposure route for phthalates, such as di-iso-butyl phthalate (DiBP), dibutyl phthalate (DBP), bis(2-ethylhexyl) phthalate (DEHP), and benzyl butyl phthalate (BBP). In this study, we aimed to estimate phthalate exposure in the diet of pregnant women and assess the health risk. A total of 517 pregnant women in the first trimester were recruited, and food frequency questionnaires were collected. A simple distribution assessment method was used to estimate daily exposure, and the hazard index (HI) method was used to assess cumulative risk. The maximum daily dietary exposure to DEHP, DBP, DiBP, and BBP was 5.25, 3.17, 2.59, and 0.58 µg/kg bw/day, respectively, and did not exceed the safety limit values. Cereals and vegetables were the main sources of the estimated daily intake (EDI) of phthalates in the diet. The cumulative risk assessment, based on the European Food Safety Authority tolerable daily intake (TDI) and the US Environmental Protection Agency reference dose (RfD), did not exceed the threshold of 1. DiBP, DBP, and DEHP had higher hazard quotient (HQ) values for cumulative health risk than BBP. In conclusion, a low health risk was posed by the cumulative dietary exposure to phthalates for pregnant women in Beijing.

Toxicokinetic assessment of inhalatory absorption of Diisobutyl phthalate (DiBP) using a novel thermal desorption-GC-MS method to determine phthalate diesters in blood plasma.

Phthalates are endocrine disrupting compounds that have been found in outdoor and indoor air. However, little is known about their inhalatory absorption. Although measurement of urinary metabolites is the current standard, complex and convergent metabolism of phthalates poses the necessity for alternative methodologies such as the quantitation of parental compounds in plasma. We determined the inhalatory absorption of Diisobutyl phthalate (DiBP) using a novel method based on a thermal desorption probe (TSP)-gas chromatography-mass spectrometry developed for the detection and quantitation of nine phthalate diesters in blood plasma, which fulfilled the acceptance criteria suggested by FDA guidelines regarding specificity, matrix effect, recovery, linearity, sensitivity, accuracy, and precision. After inhalation, plasma concentration of DiBP exhibited two peaks, suggesting a first, rapid absorption event, followed by a second, delayed one and a first order elimination stage. Half-life was calculated as 62 min and bioavailability, compared to IV route, was 15%.

Phthalic Acid Esters: Natural Sources and Biological Activities.

Phthalic acid esters (PAEs) are a class of lipophilic chemicals widely used as plasticizers and additives to improve various products' mechanical extensibility and flexibility. At present, synthesized PAEs, which are considered to cause potential hazards to ecosystem functioning and public health, have been easily detected in the atmosphere, water, soil, and sediments; PAEs are also frequently discovered in plant and microorganism sources, suggesting the possibility that they might be biosynthesized in nature. In this review, we summarize that PAEs have not only been identified in the organic solvent extracts, root exudates, and essential oils of a large number of different plant species, but also isolated and purified from various algae, bacteria, and fungi. Dominant PAEs identified from natural sources generally include di-n-butyl phthalate, diethyl phthalate, dimethyl phthalate, di(2-ethylhexyl) phthalate, diisobutyl phthalate, diisooctyl phthalate, etc. Further studies reveal that PAEs can be biosynthesized by at least several algae. PAEs are reported to possess allelopathic, antimicrobial, insecticidal, and other biological activities, which might enhance the competitiveness of plants, algae, and microorganisms to better accommodate biotic and abiotic stress. These findings suggest that PAEs should not be treated solely as a "human-made pollutant" simply because they have been extensively synthesized and utilized; on the other hand, synthesized PAEs entering the ecosystem might disrupt the metabolic process of certain plant, algal, and microbial communities. Therefore, further studies are required to elucidate the relevant mechanisms and ecological consequences.

Phthalates in children toys available in Malaysian market: Quantification and potential human health risk.

In Malaysia, inexpensive toys are sold in various urban and rural shops. Although safety regulations for toys are available in Malaysia there are limited reports about the chemicals in toys. Thus, this study aimed to assess the levels of phthalates (bis-[2-ethylhexyl] phthalate [DEHP], diethyl phthalate [DEP], diisobutyl phthalate [DiBP] and dibutyl phthalate [DBP]) in inexpensive toys sold at local markets in Kuala Lumpur (Malaysia) and its health risks to children. All 30 toys analysed exceeded the European Union limit (0.1 % by mass) indicating that the phthalate used as plasticizers is still prominent in toys. Bis-[2-ethylhexyl] phthalate (DEHP) was the highest detected phthalate in toy sample which was manufactured in Malaysia and sold without Malaysian Conformity Mark. Significant association was found between phthalate levels and country, indicating a need to monitor and raise public awareness about potential toxic chemicals in inexpensive toys and children's products. There are few inexpensive toys that have a hazard index value of more than one, which is associated with developmental toxicity and causes developmental effects in children. Given the severity and complexity of these toys to children health, there is a need for regular monitoring and effective enforcements to develop an acceptable baseline level of children toys products manufactured in or imported to Malaysia. Furthermore, risk management efforts should also include all the stakeholders involved in toy production, policy makers as well as consumers to ensure only toy products with proper labels being sole and purchased.

Transcriptomic responses predict the toxic effect of parental co-exposure to dibutyl phthalate and diisobutyl phthalate on the early development of zebrafish offspring.

Dibutyl phthalate (DBP) and diisobutyl phthalate (DiBP) have been reported to exhibit reproductive toxicity in vertebrates. However, the combined effect of DBP and DiBP on offspring of exposed parents remains unclear, especially for aquatic organisms such as fish. The aims of this study were to assess the effects of parental co-exposure to DBP and DiBP on early development of zebrafish offspring, and to explore the potential molecular mechanisms involved. The early developmental indicators and transcriptomic profiles of F1 larvae were examined after parental exposure to DBP, DiBP and their mixtures (Mix) for 30 days. Results showed that parental exposure to DBP and DiBP, alone or in combination, resulted in increased hatchability at 48 hpf and heart rate at 96 hpf, and increased the prevalence of malformations and mortality in F1 larvae. Generalized linear model (GLM) suggested an antagonistic interactive effect between DBP and DiBP on mortality and malformations of F1 larvae. The transcriptomic analysis revealed that the molecular mechanisms of parental co-exposure were different from those of either chemical alone. Disruption of molecular functions involved unfolded protein binding, E-box binding and photoreceptor activity in F1 larvae. These findings provide initial insights in the potential mechanism of action of parental co-exposure to DBP and DiBP.

Toxicokinetics of diisobutyl phthalate and its major metabolite, monoisobutyl phthalate, in rats: UPLC-ESI-MS/MS method development for the simultaneous determination of diisobutyl phthalate and its major metabolite, monoisobutyl phthalate, in rat plasma, urine, feces, and 11 various tissues collected from a toxicokinetic study.

The aim of this study was to explore the toxicokinetics of diisobutyl-phthalate (DiBP) and its major metabolite, monoisobutyl-phthalate (MiBP), by developing a UPLC-ESI-MS/MS method for simultaneously measuring DiBP and MiBP in rat plasma, urine, feces, and 11 different tissues. For the experiment, 0.1% (v/v) aqueous formic acid and acetonitrile mobile phase by gradient elution at a flow rate of 0.3 mL/min, equipped with a KINETEX core-shell C18-column (50 × 2.1 mm, 1.7 µm), was used to completely separate analytes. The mass transitions were m/z 279.1 â†’ 149.0 for DiBP, 221.0 â†’ 77.0 for MiBP, and 283.2 â†’ 153.0 for DiBP-d4 as an internal standard. The developed assay had lower limits of quantification of 0.01 ng/mL for DiBP and 0.1 ng/mL for MiBP at all biological matrices. Toxicokinetics of DiBP were characterized by extensive distribution, short half-life, and high clearance. DiBP was rapidly metabolized to MiBP, with MiBP levels consistently exceeding the DiBP levels. Distribution of MiBP to tissues was considerable. The developed analytical method satisfied international criteria and was successfully applied to toxicokinetic studies after oral and intravenous administration of DiBP to rats. Findings of this study may be useful for evaluating the external exposure and toxic potential of DiBP and its metabolite in risk assessment.

Migration regularity of phthalates in polyethylene wrap film of food packaging.

As a kind of polymer material additive, phthalic acid esters (PAEs) are widely used in food industry. However, PAEs are environmental endocrine disruptors with reproductive toxicity and teratogenic carcinogenicity, which are difficult to be degraded in the natural environment. In this paper, gas chromatography-mass spectrometer (GC-MS) methods for PAEs in polyethylene wrap film were optimized. For diisobutyl phthalate (DIBP) and dibutyl phthalate (DBP) that were mainly detected, the method had a good linearity in 1 to 500 ng/g. Then, we confirmed that the migration of DIBP and DBP from polyethylene wrap film increased with time and temperature. It is found that the migration law in different food simulations well followed the migration dynamics first-level model. The rate constant K1 and initial release rate V0 are inversely proportional to the polarity of the simulated liquid. We hope that this study can serve as a valuable reference for further research on the migration of food packing materials. PRACTICAL APPLICATION: In this paper, we present a simple example of applying migration model to evaluate the migration behaviors of PAEs in food packaging materials along with their hazardous properties. It can serve as a valuable reference for further research on the migration of food packing materials.

The reproductive toxicity and potential mechanisms of combined exposure to dibutyl phthalate and diisobutyl phthalate in male zebrafish (Danio rerio).

Dibutyl phthalate (DBP) and diisobutyl phthalate (DiBP) are phthalate compounds frequently detected in the environment. Despite increasing awareness of their toxicity in human and animals, the male reproductive toxicity of their combined exposure remains elusive. The purposes of this study were to investigate whether combined exposure to DBP and DiBP could induce male reproductive toxicity, and to explore the potential toxicological mechanisms. Adult male zebrafish were exposed to DBP (11, 113 and 1133 µg L-1), DiBP (10, 103 and 1038 µg L-1) and their mixtures (Mix) (11 + 10, 113 + 103, 1133 + 1038 µg L-1) for 30 days, and their effects on plasma hormone secretion, testis histology and transcriptomics were examined. Highest concentrations of Mix exposure caused greater imbalance ratio of T/E2 and more severe structural damage to testis than single exposure. These effects were consistent with the testis transcriptome analysis for which 4570 genes were differentially expressed in Mix exposure, while 2795 and 1613 genes were differentially expressed in DBP and DiBP, respectively. KEGG pathway analysis showed that both single and combined exposure of DBP and DiBP could affect cytokine-cytokine receptor interaction. The difference was that combined exposure could also affect steroid hormone synthesis, extracellular matrix receptor interaction, retinol metabolism, and PPAR signaling pathways. These results demonstrated that combined exposure to DBP and DiBP could disrupt spermatogenesis and elicit male reproductive toxicity in zebrafish.

Development of a whole-cell biocatalyst for diisobutyl phthalate degradation by functional display of a carboxylesterase on the surface of Escherichia coli.

BACKGROUND: Phthalic acid esters (PAEs) are widely used as plasticizers or additives during the industrial manufacturing of plastic products. PAEs have been detected in both aquatic and terrestrial environments due to their overuse. Exposure of PAEs results in human health concerns and environmental pollution. Diisobutyl phthalate is one of the main plasticizers in PAEs. Cell surface display of recombinant proteins has become a powerful tool for biotechnology applications. In this current study, a carboxylesterase was displayed on the surface of Escherichia coli cells, for use as whole-cell biocatalyst in diisobutyl phthalate biodegradation. RESULTS: A carboxylesterase-encoding gene (carEW) identified from Bacillus sp. K91, was fused to the N-terminal of ice nucleation protein (inpn) anchor from Pseudomonas syringae and gfp gene, and the fused protein was then cloned into pET-28a(+) vector and was expressed in Escherichia coli BL21(DE3) cells. The surface localization of INPN-CarEW/or INPN-CarEW-GFP fusion protein was confirmed by SDS-PAGE, western blot, proteinase accessibility assay, and green fluorescence measurement. The catalytic activity of the constructed E. coli surface-displayed cells was determined. The cell-surface-displayed CarEW displayed optimal temperature of 45 °C and optimal pH of 9.0, using p-NPC2 as substrate. In addition, the whole cell biocatalyst retained ~ 100% and ~ 200% of its original activity per OD600 over a period of 23 days at 45 °C and one month at 4 °C, exhibiting the better stability than free CarEW. Furthermore, approximately 1.5 mg/ml of DiBP was degraded by 10 U of surface-displayed CarEW cells in 120 min. CONCLUSIONS: This work provides a promising strategy of cost-efficient biodegradation of diisobutyl phthalate for environmental bioremediation by displaying CarEW on the surface of E. coli cells. This approach might also provide a reference in treatment of other different kinds of environmental pollutants by displaying the enzyme of interest on the cell surface of a harmless microorganism.

Presence, distribution and risk assessment of phthalic acid esters (PAEs) in suburban plastic film pepper-growing greenhouses with different service life.

The widespread usage of plastic film increased the content of phthalic acid esters (PAEs) in the environment, causing PAE residue in vegetables and subsequently increasing health risks to humans when consuming them. In this work, the presence, distribution and risk assessment of 15 PAEs in soils and peppers from suburban plastic film pepper-growing greenhouses were investigated. The total PAE contents in soil and pepper samples ranged from 320.1 to 971.2 µg/kg (586.3 µg/kg on average) and from 196.6 to 304.2 µg/kg (245.4 µg/kg on average), respectively. Di (2-ethyl)hexyl, dibutyl and diisobutyl phthalates (DEHP, DnBP and DiBP, respectively) were the most abundant in both soil and pepper samples. Specifically, DEHP showed the highest content in soils, while the DnBP content was the highest in peppers. The total PAE content in soils from pepper-greenhouses was much lower than in the agricultural soils mulched with plastic films, but significantly higher than in the agricultural soils from open uncovered fields. The total PAE content in peppers decreased as the service life of plastic film greenhouses increased. Correlation analysis suggested that the difference in distribution and accumulation behaviors of individual PAEs in greenhouse systems was correlated with their physicochemical properties. The non-cancer and carcinogenic risks of priority PAEs show low risks of PAEs detected in pepper and soil samples from the suburban plastic film greenhouses to human health.

Accumulation and transport patterns of six phthalic acid esters (PAEs) in two leafy vegetables under hydroponic conditions.

In this study, we investigated the accumulation and transport patterns of six phthalic acid esters (PAEs) in two leafy vegetables under hydroponic conditions. The tested PAEs included dibutyl phthalate (DBP), diethyl phthalate (DEP), diallyl phthalate (DAP), diisobutyl phthalate (DIBP), dimethyl phthalate (DMP) and benzyl butyl phthalate (BBP), and the tested vegetables included Gaogengbai and Ziyoucai. The results revealed that the six PAEs were taken up by vegetables from the solution, although their accumulation and distribution varied among PAEs. The ability of concentrating PAEs into the roots followed the order of BBP > DBP > DIBP > DAP > DEP > DMP, whereas the ability of concentrating PAEs in plant shoots had the opposite order. By analysing the fractionation of the six PAEs in vegetable roots, DMP had the largest proportion in terms of apoplastic movement, while BBP had the largest proportion in terms of symplastic movement. Correlation analyses revealed that the differences among the accumulation and distribution behaviours of the six PAEs in plant tissues were not only related to their physicochemical parameters, such as alkyl chain length and the octanol/water partition coefficient (logKow), but also related to the proportion of apoplastic and symplastic movement in the plant roots. In addition, PAEs were more readily accumulated in the Gaogengbai roots than in the Ziyoucai roots; however, the opposite trend was observed for the shoots.

Spatial distribution of phthalate esters and the associated response of enzyme activities and microbial community composition in typical plastic-shed vegetable soils in China.

The widespread use of phthalate esters (PAEs) in plastic products has made them ubiquitous in environment. In this study, 93 soil samples were collected in 31 plastic-sheds from one of China's largest vegetable production bases, Shouguang City, Shandong Province, to investigate the pollution characteristics and composition of PAEs in soils. Eleven PAEs were detected in the soil samples with the total concentration of 756-1590 µg kg-1 dry soil. Di (2-ethylhexyl) phthalate (DEHP), bis (2-n-butoxyethyl) phthalate (DBEP), di-isobutyl phthalate (DiBP) and di-n-butyl phthalate (DBP) were the main pollutants with the highest concentrations. Moreover, soil properties, including pH, total organic carbon (TOC), soil enzyme activities, and soil microbial community characteristics, were monitored to explore the associated formation mechanisms. The concentration of PAEs in the plastic-shed vegetable soils was regionalized and the contamination degree in different regions was related to soil microbial characteristics and soil enzyme activities. Phthalate ester is positively correlated with catalase and sucrase, and negatively correlated with dehydrogenase and urease. Furthermore, some tolerant and sensitive bacteria were selected, which possibly could be used as potential indicators of PAE contamination in soil. Dimethyl phthalate (DMP) and DBP also had greater effects on the soil microbial community than other PAEs. The results will provide essential data and support the control of PAEs in plastic-shed vegetable soils in China.

Phthalate esters in biota, air and water in an agricultural area of western China, with emphasis on bioaccumulation and human exposure.

Phthalate esters (PAEs) have been shown to be ubiquitous in abiotic and biotic environmental compartments; however, information about bioaccumulation behavior and human exposure, both via environmental exposure and the diet, are limited. Herein, we report the concentrations and composition profiles of phthalate esters (PAEs) in biological samples, river water, indoor air, and outdoor air samples collected from an agricultural site in western China. Dibutyl phthalate (DNBP) occupied a relatively high abundance in biological samples, discrepant with the environmental samples in which di-(2-ethylhexyl) phthalate (DEHP) was the dominant congener. Significant correlations (P < 0.05) were observed between the biota and river water samples, indicating that river water heavily influenced PAE accumulation in biological samples. The mean log Bioaccumulation Factors (BAFs) varied from 0.91 to 2.96, which implies that most PAE congeners are not likely to accumulate in organisms. No obvious trends were observed between log octanol-water partition coefficient (KOW) and log BAF values, nor between log octanol-air partition coefficient (KOW) and biota-air accumulation factors (BAAFs). Nevertheless, the calculated log air-water partitioning factors (AWPFs) of diethyl phthalate (DEP), dimethyl phthalate (DMP), and butyl benzyl phthalate (BBP) were similar to predicted values whereas those for diisobutyl phthalate (DIBP), DNBP and DEHP were significantly higher. The estimated daily intakes of PAEs via food ingestion and environmental exposure were 15, 9.4 and 1.2 ng/kg-bw/day in toddlers, children and adults, respectively, laying at the low end of the reported data and well below the reference dose.

Changes in urinary excretion of phthalates, phthalate substitutes, bisphenols and other polychlorinated and phenolic substances in young Danish men; 2009-2017.

During the past two decades human exposure to bisphenol A (BPA) and phthalates such as di-iso-butyl phthalate (DiBP), di-n-butyl phthalate (DnBP), butylbenzyl phthalate (BBzP) and di-(2-ethyl-hexyl) phthalate (DEHP) has received substantial interest due to widespread population exposures and potential endocrine disrupting effects. Therefore, these chemicals have gradually been restricted and phased out through legislation. However, humans are still exposed to a wide range of other less studied phthalates, phthalate substitutes and BPA analogues as well as other polychlorinated and phenolic substances. In this study, we investigated human exposure to these chemicals over the past decade. Three hundred urine samples collected in 2009, 2013 and 2017 (100 samples each year) from young Danish men of the general population, participating in a large on-going cross-sectional study, were selected for the present time trend study. The urinary concentration of metabolites of 15 phthalates, di-2-ethylhexyl terephthalate (DEHTP) and di-iso-nonyl-cyclohexane-1,2-dicarboxylate (DINCH), seven bisphenols including BPA, bisphenol S (BPS) and bisphenol F (BPF), as well as triclosan, triclocarban, benzophenone-3, three chlorophenols and two phenylphenols were analyzed by two new sensitive LC-MS/MS methods developed and validated for the present study. A significant decrease in urinary concentrations over time was observed for the majority of the chemicals. Median concentrations of BPA and the metabolites of DiBP, DnBP, BBzP and DEHP were more than halved from 2009 to 2017. Similar decreases were observed for triclosan and the chloro- and phenylphenols. In contrast, metabolites of the two phthalate substitutes DEHTP and DINCH increased more than 20 and 2 times, respectively. The potential BPA substitutes; BPS and BPF also increased, but only slightly. Despite these new exposure patterns, the exposure to the old well-known chemicals, such as DiBP, DnBP, BBzP, DEHP and BPA was still higher in 2017 compared to the exposure level of the new substitutes such as DEHTP, DINCH, BPS and BPF. A significant decrease in internal exposure to most of the common phthalates and BPA over the past decade was observed, reflecting market changes and regulatory measures implemented in EU. Despite increasing exposures to some of the known phthalate substitutes and BPA analogues, the total amount of each measured chemical group (original and substitute analytes combined) was lower in the more recently collected samples. This indicates only partial direct substitution or substitution by chemicals not covered in this approach, or a general decline in the exposure to these chemical/product groups over the last decade.

Transcriptomic analysis reveals potential mechanisms of toxicity in a combined exposure to dibutyl phthalate and diisobutyl phthalate in zebrafish (Danio rerio) ovary.

Phthalate esters (PAEs), which are notable plasticizers, can be prolific contaminants in aquatic environments, and have been shown to induce reproductive toxicity. However, the studies concerning their toxicity towards aquatic species are based on individual chemicals, and the combined toxicity of PAEs to aquatic organisms remains unclear. The aim of this study was to explore the potential toxicity mechanisms associated with combined exposure to dibutyl phthalate (DBP) and diisobutyl phthalate (DiBP) in adult female zebrafish ovaries. Zebrafish were exposed to DBP, DiBP and their mixtures for 30 days, and their effects on ovarian histology, plasma sex hormones and ovarian transcriptomics were investigated. Plasma estradiol (E2) levels were significantly decreased by 38.9% in the DBP-1133 exposure group and 41.0% in the DiBP-1038 exposure group. The percentage of late/mature oocytes was also significantly decreased by 17.3% under DBP-1133 exposure and 16.2% under DiBP-1038 exposure, while that under combined exposure was not significantly affected. Nevertheless, transcriptome sequencing revealed 2564 differentially expressed genes (DEGs) in zebrafish ovaries after exposure to the mixtures. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis showed that the DEGs were involved in the neuroactive ligand-receptor interaction, GnRH, progesterone-mediated oocyte maturation, oocyte meiosis and steroid hormone biosynthesis signaling pathways. These results revealed that combined exposure exerts potential reproductive toxicity at the molecular level.

Acute effects on human sperm exposed in vitro to cadmium chloride and diisobutyl phthalate.

Epidemiological studies reported a negative relationship between concentrations of heavy metals and phthalates in seminal fluid and semen quality, likely compromising male fertility potential. The aim of this study was to investigate the in vitro effects of cadmium chloride (CdCl2), a common heavy metal, and diisobutyl phthalate (DIBP), a common phthalate ester, on human sperm functions necessary for fertilization. After in vitro incubation of spermatozoa with 10 µM CdCl2 or 100 and 200 µM DIBP for 24 h, a significant decrease of sperm progressive and hyperactivated motility was observed. The exposure to each of the two toxic agents also induced spontaneous sperm acrosome reaction and blunted the physiological response to progesterone. Both agents induced an increase of caspase activity suggesting triggering of an apoptotic pathway. Our results suggest that acute exposure of spermatozoa to these pollutants may impair sperm ability to reach and fertilize the oocyte.