PCB Sulfates in Serum from Mothers and Children in Urban and Rural U.S. Communities.

Serum samples from 24 subjects (6 mother-daughter and 6 mother-son dyads) in a rural community (Columbus Junction, Iowa) and 24 subjects (6 mother-daughter and 6 mother-son dyads) in an urban community (East Chicago, Indiana) were analyzed for 74 sulfated metabolites of polychlorinated biphenyls (PCBs). We detected significantly higher mean concentrations of total assessed PCB sulfates in the urban group (110-8900 ng/g fresh weight of serum, mean = 3400 ng/g, standard error = 300) than in the rural cohort (530-6700 ng/g fresh weight of serum, mean = 1800 ng/g, standard error = 500). Eight PCB sulfate congeners (4-PCB 2 sulfate, 4'-PCB 2 sulfate, 2'-PCB 3 sulfate, 4'-PCB 3 sulfate, 4-PCB 11 sulfate, 4'-PCB 18 sulfate, 4'-PCB 25 sulfate, and 4-PCB 52 sulfate) contributed over 90% of the total assessed PCB sulfates in most individuals. The serum samples were enriched in PCB sulfates with fewer than 5 chlorine atoms, and this congener distribution differed from those of PCBs and hydroxylated PCBs in previous studies in the same communities. Regression analysis indicated several significant congener-specific correlations in mother-child dyads, and these relationships differed by location and by mother-daughter or mother-son dyads. This is the first study reporting a broad range of PCB sulfates in populations from urban and rural areas.

Hydroxylated Polychlorinated Biphenyls Are Emerging Legacy Pollutants in Contaminated Sediments.

We measured the concentrations of 837 hydroxylated polychlorinated biphenyls (OH-PCBs, in 275 chromatographic peaks) and 209 polychlorinated biphenyls (PCBs, in 174 chromatographic peaks) in sediments from New Bedford Harbor in Massachusetts, Altavista wastewater lagoon in Virginia, and the Indiana Harbor and Ship Canal in Indiana, USA and in the original commercial PCB mixtures Aroclors 1016, 1242, 1248, and 1254. We used the correlation between homologues and the peak responses to quantify the full suite of OH-PCBs including those without authentic standards available. We found that OH-PCB levels are approximately 0.4% of the PCB levels in sediments and less than 0.0025% in Aroclors. The OH-PCB congener distributions of sediments are different from those of Aroclors and are different according to sites. We also identified a previously unknown compound, 4-OH-PCB52, which together with 4'-OH-PCB18 made up almost 30% of the OH-PCBs in New Bedford Harbor sediments but less than 1.2% in the Aroclors and 3.3% in any other sediments. This indicates site-specific environmental transformations of PCBs to OH-PCBs. We conclude that the majority of OH-PCBs in these sediments are generated in the environment. Our findings suggest that these toxic breakdown products of PCBs are prevalent in PCB-contaminated sediments and present an emerging concern for humans and ecosystems.

Interconversion between methoxylated, hydroxylated and sulfated metabolites of PCB 3 in whole poplar plants.

Methoxylated polychlorinated biphenyls (MeO-PCBs) are overlooked metabolites of PCBs. In general, they are more toxic to plants than their parent congeners. However, information on the fate of MeO-PCBs and the relationship between methoxylated, hydroxylated and sulfated metabolites of PCBs in plants is scarce. In this work, poplar plants (Populus deltoides × nigra, DN34) were hydroponically and separately exposed to 4'-methoxy-4-monochlorobiphenyl (4'-MeO-PCB 3) and 4'-PCB 3 sulfate for 10 days to investigate the uptake, translocation and metabolism of MeO-PCBs and the relationship between methoxy-PCBs, hydroxyl-PCBs and PCB sulfates within plants. Results showed that 4'-MeO-PCB 3 and 4'-PCB 3 sulfate were taken up by the roots of poplar plants and translocated from roots to shoots and leaves. 4'-OH-PCB 3 and 4'-PCB 3 sulfate were identified as the hydroxylated metabolite and sulfate metabolite of 4'-MeO-PCB 3 in poplar, respectively. In the backward reaction, 4'-OH-PCB 3 and 4'-MeO-PCB 3 were found as metabolites of 4'-PCB 3 sulfate. For exposure groups, the yields of 4'-OH-PCB 3 produced from 4'-MeO-PCB 3 and 4'-PCB 3 sulfate were 1.29% and 0.13% respectively. The yield of 4'-PCB 3 sulfate which originated from 4'-MeO-PCB 3 in wood and root samples of exposure groups was only 0.02%. Only 0.04% of the initial mass of 4'-PCB 3 sulfate was transformed to 4'-MeO-PCB 3 in the exposure groups. The sulfation yield of 4'-OH-PCB 3 was higher than hydrolysis yield of 4'-PCB 3 sulfate, indicating that formation of PCB sulfates was predominant over the reverse reaction, the formation of hydroxy-PCBs. These results provide new perspective on the transport, metabolism, and fate of MeO-PCBs, and also help to better understand sources of OH-PCBs and PCB sulfates in the environment. This study provides the first evidence of interconversion of sulfate metabolites from methoxy-PCBs and methoxy-PCBs from PCB sulfates.

Detection and Quantification of Polychlorinated Biphenyl Sulfates in Human Serum.

Polychlorinated biphenyls (PCBs) are persistent toxic chemicals with both legacy sources (e.g., Aroclors) and new sources (e.g., unintentional contaminants in some pigments and varnishes). PCB sulfates are derived from further metabolism of hydroxylated PCBs (OH-PCBs), which are oxidative metabolites of PCBs. While OH-PCBs and PCB sulfates are implicated in multiple toxicological effects, studies of PCB sulfates in human serum have been limited by available analytical procedures. We have now developed a method for extraction of PCB sulfates from serum followed by differential analysis with, and without, sulfatase-catalyzed hydrolysis to OH-PCBs. A sulfatase from Helix pomatia was purified by affinity chromatography, and it displayed broad specificity for PCB sulfates without contaminant glucuronidase activity. Following sulfatase-catalyzed hydrolysis of the PCB sulfates extracted from serum, the corresponding OH-PCBs were derivatized to methoxy-PCBs and quantitated by GC-MS/MS. In a pooled sample of human serum, we identified 10 PCB sulfates, with three PCB sulfate congeners exhibiting the highest concentrations from 1200 to 3970 pg/g of serum. In conclusion, we have developed a sensitive and specific method for the determination of PCB sulfates in human serum.

Polychlorinated Biphenyls in Food.

We measured the concentrations of 205 polychlorinated biphenyl (PCB) congeners in 26 food items: beef steak, butter, canned tuna, catfish, cheese, eggs, french fries, fried chicken, ground beef, ground pork, hamburger, hot dog, ice cream, liver, luncheon meat, margarine, meat-free dinner, milk, pizza, poultry, salmon, sausage, shrimp, sliced ham, tilapia, and vegetable oil. Using Diet History Questionnaire II, we calculated the PCB dietary exposure in mothers and children participating in the AESOP Study in East Chicago, Indiana, and Columbus Junction, Iowa. Salmon had the highest concentration followed by canned tuna, but fish is a minor contributor to exposure. Other animal proteins are more important sources of PCB dietary exposure in this study population. Despite the inclusion of few congeners and food types in previous studies, we found evidence of a decline in PCB concentrations over the last 20 years. We also found strong associations of PCB congener distributions with Aroclors in most foods and found manufacturing byproduct PCBs, including PCB11, in tilapia and catfish. The reduction in PCB levels in food indicates that dietary exposure is comparable to PCB inhalation exposures reported for the same study population.

A semi-target analytical method for quantification of OH-PCBs in environmental samples.

Hydroxylated polychlorinated biphenyls (OH-PCBs) are oxidative metabolites of PCBs and residuals found in original Aroclors. OH-PCBs are known to play a role as genotoxicants, carcinogens, and hormone disruptors, and therefore it is important to quantify their presence in human tissues, organisms, and environmental matrices. Of 837 possible mono-OH-PCBs congeners, there are only ~ 70 methoxylated PCB (MeO-PCB) standards commercially available. Hence, a semi-target analytical method is needed for unknown OH-PCBs. The mass concentrations of these unknowns are sometimes determined by assuming the peak responses of other available compounds. This can bias the results due to the choices and availabilities of standards. To overcome this issue, we investigated the peak responses of all commercially available MeO-PCB standards with gas chromatography (GC) coupling with triple quadrupole (QqQ) mass spectrometry (MS) system, with positive electron impact (EI) ionization at 20-70 eV in selected ion monitoring (SIM) mode. We found correlations between the relative peak responses (RRFs) and the number of chlorine (#Cl) in the molecules of MeO-PCBs. Among the studied models, the quadratic regression of #Cl is the most suitable model in the RRF prediction (RRF = ß1 × #Cl^2 + ß0) when the peak responses are captured at 30 eV. We evaluated the performance of the model by analyzing 12 synthesized MeO-PCB standards and a PCB-contaminated sediment collected from a wastewater lagoon. We further demonstrate the utility of the model using a different chromatography column and GC-EI-MS system. We found the method and associated model to be sufficiently simple, accurate, and versatile for use in quantifying OH-PCBs in complex environmental samples.

The emerging contaminant 3,3'-dichlorobiphenyl (PCB-11) impedes Ahr activation and Cyp1a activity to modify embryotoxicity of Ahr ligands in the zebrafish embryo model (Danio rerio).

3,3'-dichlorobiphenyl (PCB-11) is an emerging PCB congener widely detected in environmental samples and human serum, but its toxicity potential is poorly understood. We assessed the effects of three concentrations of PCB-11 on embryotoxicity and Aryl hydrocarbon receptor (Ahr) pathway interactions in zebrafish embryos (Danio rerio). Wildtype AB or transgenic Tg(gut:GFP) strain zebrafish embryos were exposed to static concentrations of PCB-11 (0, 0.2, 2, or 20 µM) from 24 to 96 h post fertilization (hpf), and gross morphology, Cytochrome P4501a (Cyp1a) activity, and liver development were assessed via microscopy. Ahr interactions were probed via co-exposures with PCB-126 or beta-naphthoflavone (BNF). Embryos exposed to 20 µM PCB-11 were also collected for PCB-11 body burden, qRT-PCR, RNAseq, and histology. Zebrafish exposed to 20 µM PCB-11 absorbed 0.18% PCB-11 per embryo at 28 hpf and 0.61% by 96 hpf, and their media retained 1.36% PCB-11 at 28 hpf and 0.84% at 96 hpf. This concentration did not affect gross morphology, but altered the transcription of xenobiotic metabolism and liver development genes, impeded liver development, and increased hepatocyte vacuole formation. In co-exposures, 20 µM PCB-11 prevented deformities caused by PCB-126 but exacerbated deformities in co-exposures with BNF. This study suggests that PCB-11 can affect liver development, act as a partial agonist/antagonist of the Ahr pathway, and act as an antagonist of Cyp1a activity to modify the toxicity of compounds that interact with the Ahr pathway.

Chemistry of ecteinascidins. Part 3: preparation of 2'-N-acyl derivatives of ecteinascidin 770 and evaluation of cytotoxicity.

A three-step transformation of ecteinascidin 770 (1b) into 2'-N-indole-3-carbonyl derivative 3 via 18,6'-O-bisallyl-protected derivative 4a, which was shown to have higher cytotoxicity than 1b, is presented. In addition, a number of 2'-N amide derivatives of 1b have been prepared from 4a and their in vitro cytotoxicity were determined by measuring IC50 values against human cell lines HCT116, QG56, and DU145. Benzoyl amide derivatives 7a-c showed similar in vitro cytotoxicity to 1b, whereas the nitrogen-containing heterocyclic derivatives 7d-h and cinnamoyl derivatives 9a-b showed higher cytotoxicity than 1b. In contrast, the 18,6'-O-bisallyl protected derivatives 4a-c, 6a-h, and 8a-b showed dramatic decreases in cytotoxicity relative to 1b.