Isolation of Two Plasticizers, Bis(2-ethylhexyl) Terephthalate and Bis(2-ethylhexyl) Phthalate, from Capparis spinosa L. Leaves.

Many plants have been known to be contaminated and accumulate plasticizers from the environment, including water sources, soil, and atmosphere. Plasticizers are used to confer elasticity and flexibility to various fiber and plastic products. Consumption of plasticizers can lead to many adverse effects on human health, including reproductive and developmental toxicity, endocrine disruption, and cancer. Herein, we report for the first time that two plasticizers, bis(2-ethylhexyl) terephthalate (DEHT) and bis(2-ethylhexyl) phthalate (DEHP), have been isolated from the leaves of Capparis spinosa L. (the caper bush), a plant that is widely used in food seasonings and traditional medicine. 297 mg/kg of DEHT and 48 mg/kg of DEHP were isolated from dried and grounded C. spinosa L. leaves using column chromatography and semi-preparative high-performance liquid chromatography. Our study adds to the increase in the detection of plasticizers in our food and medicinal plants and to the alarming concern about their potential adverse effects on human health.

[Determination and Traceability Analysis of Phthalic Acid Esters in Garlic (Allium stivum L.) from Jiangsu Province, China].

Phthalic acid esters (PAEs) are ubiquitous environmental pollutants and are recognized as a threat to the environment and agricultural product safety across the world. In order to investigate the level of PAEs in garlic, soils, and agricultural films from Pizhou City, Jiangsu province, China, 11 garlic samples, 106 soil samples, and 4 agricultural film samples were collected and analyzed using GC-MS. In addition, the uptake and transport characteristics of six PAEs compounds classified as priority pollutants by the United States Environmental Protection Agency (EPA) in the garlic cultivar Daqingke were investigated under hydroponic conditions. The results indicated that dibutyl phthalate (DBP) and di-(2-ethylhexyl) phthalate (DEHP) were the dominant PAEs species in garlic cloves of the different garlic varieties from Pizhou City. The average contents of DBP and DEHP in garlic cloves were 0.611 mg·kg-1 and 0.167 mg·kg-1, respectively, which were significantly higher than those of the commercial varieties of garlic. The concentrations of DBP and DEHP differed in three tissues of garlic bulbs, ordered as the skin of garlic bulb>skin of garlic clove>garlic clove. Dimethyl phthalate (DMP), diethyl phthalate (DEP), diisobutyl phthalate (DIBP), DBP, and DEHP were the main PAEs species and were detected in all the surface soils collected from Pizhou City. Compared with the soil allowable concentrations of the six PAEs in the United States, the DMP and DBP concentrations in approximately 100% and 63.2% of soil samples exceeded the recommended allowable concentrations set by the EPA. However, the levels of DEP, DIBP, and DEHP in the soils were below the maximum allowable concentrations set by the EPA. Nevertheless, the average content of DEHP in soils was 486 µg·kg-1 and was found to be much higher than that in the other four PAEs. Six PAEs, including DMP, DEP, DIBP, DBP, butyl benzyl phthalate (BBP), and DEHP, were detected in all the agricultural film samples. Among them, the contents of DBP and DEHP in the agricultural films were the highest, accounting for 53.7%-63.2% of the total PAEs. The amount of PAEs present in the residual film was significantly lower than that in the new film, and all six PAEs were detected in garlic or soil samples, suggesting that agricultural film can be an important source of PAEs in garlic farming soils and garlic. Furthermore, the garlic plants absorbed DMP and DEP efficiently from the substrate and showed higher translocation factors (TFs) for DMP and DEP than those for DBP, BBP, DEHP, and di-n-octyl phthalate (DnOP), resulting in a higher accumulation of DMP and DEP in the over-ground parts of garlic. In contrast, DBP and BBP in roots of garlic displayed higher bioconcentration factors (57.4 and 81.5, respectively) compared to those of the other four PAEs, whereas the TFs of DBP and BBP were lower; this may have contributed to the high accumulation of DBP in garlic bulbs. The BCFs and TFs of DEHP and DnOP in garlic were relatively lower, but the DEHP had been detected in all garlic cloves, which may be a result of the higher DEHP contents in soils.

Occurrence and distribution of phthalate esters and microplastics in wastewater treatment plants in Taiwan and their toxicological risks.

Phthalate esters (PAEs) are fat soluble synthetic chemicals, usually regarded as plasticizers for being added in numerous plastic products. Thus, environmental and health hazards of PAEs are associated with increasing plastic pollution. In this study, PAEs from sludge samples collected from water, sewage, and industrial treatment plants (N = 17) were analyzed using Gas Chromatography/Mass Spectrometry. Microplastics (MPs) were also quantified and correlated with PAEs. Results showed the highest average PAE concentrations in sewage treatment plants. The greatest ΣPAEs concentration were found in sewage treatment plant (STP4) with 32,414 µg/kg dw, while the lowest found in water treatment plant (WTP3) with 2062 µg/kg dw. Among different PAEs, di-(2-ethyl hexyl) phthalate (DEHP) contributes the highest. Similarly, DEHP, di-n-octyl phthalate (DnOP) and diisononyl phthalate (DiNP) significantly correlated with the total PAEs indicating their large contribution to sludge contamination. The abundance of microplastics in sludge ranged between 1 and 7 MP/g, highest at ITP6, but not detected in some stations. While microplastics may potentially increase PAEs, there was no significant relationship between ΣPAEs and MP abundance. The estimated human daily intake of DEHP and di-n-butyl phthalate (DnBP) when contaminated sludge be used showed low toxicological risks to exposed adults. This research presents the sludge characteristics, PAEs, and microplastic concentrations in different wastewater treatment plants in Taiwan. PAE contamination was highly contributed by domestic and industrial wastes shown by their significant amounts in STP and ITP. Results further provide evidence for potential sludge recycling (WTP sludge) and application to soil.

Biodegradation patterns of the endocrine disrupting pollutant di(2-ethyl hexyl) phthalate by Fusarium culmorum.

Di(2-ethylhexyl) phthalate (DEHP) is a widely used plasticizer, which is considered an endocrine disrupting pollutant. Growth kinetics and esterases activity by biochemical tests and polyacrylamide gel electrophoresis were characterized for Fusarium culmorum grown in DEHP-supplemented (1000 mg/L) medium as the only carbon source and in control medium with glucose. Intermediate compounds of biodegraded DEHP were identified by GC-MS. F. culmorum degraded 92% of DEHP within 36 h. DEHP was degraded to butanol, hexanal, catechol and acetic acid. It is suggested that the first two compounds would transform into butanediol and the last two would enter into the Krebs cycle and would be mineralized to CO2 and H2O. DEHP induced eight esterase isoforms, which were different to those constitutive isoforms produced in the control medium. It is suggested that five enzymes (25.7, 29.5, 31.8, 97.6 and 144.5 kDa) detected during the first 36 h be involved in the primary biodegradation of DEHP. The rest of the enzymes (45.9, 66.6 and 202.9 kDa) might be involved in the final steps for DEHP metabolism. F. culmorum has a promising practical application in the treatment of DEHP-contaminated environments because it can secrete specific esterase to breakdown high concentrations of DEHP in a short period of time. This research represents the first approach for the study of esterase involved in the DEHP degradation by fungi using this phthalate as the sole source of carbon and energy.

Ameliorative effect of ginseng extract on phthalate and bisphenol A reprotoxicity during pregnancy in rats.

Phthalates (such as DEHP) and bisphenol A (BPA) are widely used chemicals in plastics manufacturing and exert public health concerns as endocrine disrupters. This study was designed to investigate the deleterious effect of DEHP and BPA on endocrine profile of pregnant female rats and the combined treatment with ginseng extract (Panax ginseng). Seventy-two pregnant rats were divided into six groups (control, ginseng, DEHP, BPA, Gin + DEHP, and Gin + BPA), 12 females per each group. The drugs were supplemented from pregnancy day 0 until day 20. Determination of serum sex hormones (testosterone, progesterone, and estradiol) were determined on days 4, 10, and 20 of pregnancy. mRNA transcripts of STAR, HSD17B3, CYP17, AKT1, and PTEN were relatively quantified against ACTB in the ovary and placenta of days 10 and 20 pregnant females by relative quantitative polymerase-chain reaction (RQ-PCR). DEHP and BPA significantly decreased the endocrine profile of testosterone, progesterone, and estradiol of days 10 and 20 of pregnant females. Combined administration of these chemicals along with ginseng extracts has returned the hormones to normal levels when compared with the control group. The ovarian and placental CYP17 and HSD17B3 mRNA transcripts showed variable expression pattern in both tissues and they were significantly affected by DEHP and BPA administration, concomitantly correlating to STAR, AKT1, PTEN, progesterone, and testosterone levels on pregnancy days 10 and 20. The results confirm the reprotoxicity of DEHP and BPA as endocrine disruptors and indicate that ginseng could be used to alleviate the toxic effects of these chemicals.

Determination of pesticide and phthalate residues in tea by QuEChERS method and their fate in processing.

In this study, the quick, easy, cheap, effective, rugged, and safe (QuEChERS) method was applied for the analysis of the multiclass pesticide residues of 12 organochlorines (OCs), 9 organophosphates (OPs), 11 synthetic pyrethroids (SPs), 4 herbicides, 6 phthalates in raw tea (loose tea, branded tea and herbal tea), and tea infusion in 4 different containers (glass cup, earthen cup, plastic bag and disposal cup). In loose tea and branded tea residues, malathion (0.257 and 0.118 mg kg-1), cypermethrin (0.065 and 0.030 mg kg-1), and fenvalerate (0.032 and 0.030 mg kg-1) were detected, respectively. In herbal tea, residues of only cypermethrin (0.053 mg kg-1) and fenvalerate (0.045 mg kg-1) were detected. Tea infusion samples contained in a plastic bag were found to be contaminated with only dibutyl phthalate (DBP) (0.038 mg kg-1). Disposable cup was found to be contaminated with DBP (0.026 mg kg-1) and diethyl phthalate (DEP) (0.004 mg kg-1). Further, to know the processing behavior of pesticides, the spiked raw tea was subjected to tea infusion at different brewing times (2, 5, 10 min). The analysis demonstrated that dimethoate, dichlorvos, and malathion had shown more than 10 % of translocation at 5 min of brewing time. Further brewing for 10 min revealed the reduction in concentration of pesticides. Leaching of phthalate residues from different plastic containers was also studied at 10, 30, and 60 min. DBP, benzyl butyl phthalate (BzBP), and di-2-(ethylhexyl) phthalate (DEHP) were leached in the tea infusion samples packed in plastic bags. On the other hand, in disposable cups, leaching of DBP, DEP, and dimethyl phthalate were found. The concentration of phthalate residues increased with retention time. Pesticide and phthalate contaminants were recorded at low quantities in few samples only.

Plasticizers excreted in urine: indication of autologous blood transfusion in sports.

BACKGROUND: Misuse of autologous blood transfusions in sports remains undetectable. The metabolites of the plasticizer di-(2-ethylhexyl)phthalate (DEHP) were recently proposed as markers of blood transfusion, based on high urinary concentrations of these compounds observed in patients subjected to blood transfusion. This study evaluates DEHP metabolites in urine for detecting autologous blood transfusion. STUDY DESIGN AND METHODS: One blood bag was drawn from moderately trained subjects and the red blood cells (RBCs) were reinfused after different storage periods. Group 1 (12 subjects) was reinfused after 14 days, and Group 2 (13 subjects), after 28 days of storage. Urine samples were collected before and after reinfusion for determination of the concentrations of three DEHP metabolites, mono-(2-ethylhexyl)phthalate, mono-(2-ethyl-5-hydroxyhexyl)phthalate, and mono-(2-ethyl-5-oxohexyl)phthalate. RESULTS: Concentrations of DEHP metabolites on the days before reinfusion were in agreement with those described after common environmental exposure. A few hours after the reinfusion a significant increase was observed for all metabolites in all volunteers. Concentrations 1 day later were still higher (p < 0.05) than before reinfusion. Variations in urine dilution supported normalization by specific gravity. Concentrations of DEHP metabolites tended to be higher after longer storage times of RBCs. CONCLUSION: Autologous transfusion with RBCs stored in plastic bags provokes an acute increase in the urinary concentrations of DEHP metabolites, allowing the detection of this doping malpractice. The window of detection is approximately 2 days. The method might be applied to urine samples submitted for antidoping testing.

Compatibility and stability of aplidine, a novel marine-derived depsipeptide antitumor agent, in infusion devices, and its hemolytic and precipitation potential upon i.v. administration.

Aplidine is a novel marine-derived antitumor agent isolated from the Mediterranean tunicate Aplidium albicans. The compound is pharmaceutically formulated as a lyophilized product containing 500 microg active substance per dosage unit. Prior to i.v. administration it is reconstituted with a solution composed of Cremophor EL, ethanol absolute and Water for Injection (15/15/70% v/v/v) with further dilution in 0.9% w/v sodium chloride for infusion (normal saline). The aim of this study was to investigate the compatibility of aplidine infusion solutions with polyvinyl chloride (PVC)-containing and PVC-free administration sets, and to determine the stability of aplidine after reconstitution and further dilution in infusion solutions. Furthermore, in vitro biocompatibility studies to estimate the hemolytic and precipitation potential of aplidine infusion solutions upon i.v. administration were conducted. In this study we show that sorption of aplidine to PVC and to a lesser extent to PVC-free administration set materials occurs. Also, most probably due to the presence of Cremophor EL in the infusion solution, significant leaching of diethylhexyl phtalate (DEHP) from the PVC administration set occurs. After reconstitution and dilution the drug is stable for at least 24 and 48 h, respectively, in glass containers when stored at room temperature (20-25 degrees C) and ambient light conditions. We found that aplidine should be administered in infusion concentrations equal or above 28.8 microg/ml using a PVC-free administration set consisting of a glass container and PVC-free infusion tubing. After reconstitution it must be diluted further with normal saline within 24 h after preparation and subsequently administered to the patient within 48 h. Additionally, results from the biocompatibility studies show that neither hemolysis nor precipitation of aplidine is expected upon i.v. administration.

Determination by gas-liquid chromatography of trace amounts of soft polyvinyl chloride plastic additives in aqueous solutions. II. Di(2-ethylhexyl) phthalate, epoxidized vegetable oils and stearates.

A method for the determination of di(2-ethylhexyl) phthalate (DEHP), epoxidized vegetable oils (EVO) and stearates in aqueous solutions is described. A stepwise extraction procedure is employed to separate DEHP and EVO from the stearates, using n-hexane as extraction solvent. EVO is transesterified to methyl esters by sodium methoxide in methanol. The stearates are derivatized by methanol containing sulfuric acid. The alkyl esters are analyzed by gas-liquid chromatography, using 3% OV-210 as the stationary phase. The concentrations of the weakest standard solutions correspond to 10, 5 and 8 micrograms/l (ppb) of DEHP, EVO and stearates, respectively, in the aqueous samples. The method is used for the determination of DEHP, EVO and stearates in intravenous solutions stored in flexible polyvinyl chloride bags.

The influence of product factors on the migration in soft polyvinyl chloride infusion fluid bags.

The concentration of epoxidized vegetable oils (EVO) and di(2-ethylhexyl) phthalate (DEHP) in infusion fluids in soft polyvinyl chloride (PVC) bags has been determined by gas-liquid chromatography prior to and after agitation. Normal saline, isotonic glucose and Ringer acetate from four different manufacturers were investigated. Agitation resulted in an increased concentration of EVO and DEHP. Significant differences in contamination level were observed between solutions of different compositions and in synonymous preparations from different manufacturers. The results indicated that the total amount of migrating EVO and DEHP was affected by the pH of the solution and by the age of the plastic material. It is suggested that the migration of EVO and DEHP during agitation is influenced by the concentration of plastic degradation products.

The influence of mono(2-ethylhexyl) phthalate and stearates on the migration in soft polyvinyl chloride infusion fluid bags.

The influence of mono(2-ethylhexyl) phthalate (MEHP) and stearates on the migration of di(2-ethylhexyl) phthalate (DEHP) and epoxidized vegetable oils (EVO) in soft polyvinyl chloride infusion fluid bags during agitation has been investigated by gas-liquid chromatography. Normal saline, isotonic glucose and Ringer acetate from 3 different manufacturers were investigated. A covariation was observed between the MEHP concentration in the solution and the pH. The MEHP concentration was not influenced by agitation. There was no correlation between the MEHP concentration in the solution and the migration of EVO and DEHP, or between the stearate concentration and the EVO and DEHP migration.

Compatibility of plastic infusion devices with diluted N-methylformamide and N,N-dimethylacetamide.

Leaching of the plasticizer di-2-ethylhexyl phthalate (DEHP) from i.v. sets and polyvinyl chloride bags was studied with solutions of a new anticancer drug, N-methylformamide (NMF), and N,N-dimethylacetamide (DMA), a structurally related solvent used in the formulation of some anticancer drugs. Solutions of 0.9% sodium chloride, 5 and 10% DMA in 0.9% sodium chloride, and 20, 40, and 60% NMF in 0.9% sodium chloride were used to fill i.v. administration sets ( Venoset -60, Abbott Laboratories) and polyvinyl chloride bags ( Viaflex -500, Travenol Laboratories). The solutions were kept in the containers for 20 hours at ambient temperature. Simulated infusions were also studied using 0, 20, and 40% NMF plus 5 and 10% DMA in 0.9% sodium chloride; both glass bottles and Viaflex -500 bags were used with the i.v. administration sets. DEHP was assayed using gas chromatography. Very small or unmeasurable amounts of DEHP were leached into 0.9% sodium chloride and 5 or 10% DMA solutions during 20-hour storage. DEHP concentrations increased exponentially with increasing NMF concentrations after 20-hour storage in both the i.v. sets and bags. Similar results were found in the simulated-infusion portion of the study. DEHP extraction from i.v. sets and polyvinyl chloride bags containing NMF and DMA in 0.9% sodium chloride is related directly to the concentrations of these organic compounds. The amount of DEHP leached into solutions is important only for solutions containing 60% NMF.