Persistent organic pollutants in red-crowned cranes (Grus japonensis) from Hokkaido, Japan.

The red-crowned crane (Grus japonensis) from eastern Hokkaido is classified as a Special Natural Monument in Japan. In this study, we determined the concentrations of persistent organic pollutants (POPs) in red-crowned crane muscle tissues (n = 47). Polychlorinated biphenyls (PCBs) had the highest median concentration (240ng/g lipid weight), followed by dichlorodiphenyltrichloroethane and its metabolites (DDTs) (150ng/g lipid weight), chlordane-related compounds (CHLs) (36ng/g lipid weight), hexachlorobenzene (HCB) (16ng/g lipid weight), hexachlorocyclohexanes (HCHs) (4.4ng/g lipid weight), polybrominated diphenyl ethers (PBDEs) (1.8ng/g lipid weight), and finally, Mirex (1.5ng/g lipid weight). Additionally, a positive correlation was found among POP concentrations. No sex differences beyond body parameters were observed. Additionally, red-crowned cranes exhibited a high enantiomeric excess of (+)-alpha-HCH, with enantiomer fractions varying from 0.51 to 0.87 (average: 0.69).

Size Distribution of Chlorinated Polycyclic Aromatic Hydrocarbons in Atmospheric Particles.

The particle size distribution of chlorinated polycyclic aromatic hydrocarbons (ClPAHs) in particulate matter (PM) in Japan is examined for the first time. PM was collected using a PM0.1 air sampler with a six-stage filter. PM was collected in October 2014 and January 2015 to observe potential seasonal variation in the atmospheric behavior and size of PM, including polycyclic aromatic hydrocarbons (PAHs) and ClPAHs. We found that the concentration of PAHs and ClPAHs between 0.5-1.0 µm and 1.0-2.5 µm markedly increase in January (i.e., the winter season). Among the ClPAHs, 1-ClPyrene and 6-ClBenzo[a]Pyrene were the most commonly occurring compounds; further, approximately 15% of ClPAHs were in the nanoparticle phase (<0.1 µm). The relatively high presence of nanoparticles is a potential human health concern because these particles can easily be deposited in the lung periphery. Lastly, we evaluated the aryl hydrocarbon receptor (AhR) ligand activity of PM extracts in each size fraction. The result indicates that PM < 2.5 µm has the strong AhR ligand activity.

Detection of antibiotics in chicken eggs obtained from supermarkets in Ho Chi Minh City, Vietnam.

The residual levels of antibiotics in Vietnamese eggs were monitored from 2014 to 2015. A total of 111 egg packages, distributed by 11 different companies, were collected from supermarkets in Ho Chi Minh City and the levels of 28 antibiotics were analyzed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) screening method. Sixteen samples tested positive for antibiotics; a total of eight compounds (enrofloxacin, ciprofloxacin, norfloxacin, sulfadimethoxine, sulfamethazine, sulfamonomethoxine, tilmicosin and trimethoprim) were detected. Enrofloxacin was detected in eight samples, with two samples exhibiting concentrations exceeding 1,000 µg kg-1. Tilmicosin was detected in three samples at a range of 49-568 µg kg-1. We observed that two of the 11 companies frequently sold antibiotic-contaminated eggs (detection rates of 56 and 60%), suggesting that a number of companies do not regulate the use of antibiotics in egg-laying hens. Our findings indicate that livestock farmers require instruction regarding antibiotic use and that continual antibiotic monitoring is essential in Vietnam.

Identification and characterization of oxidative metabolites of 1-chloropyrene.

Polycyclic aromatic hydrocarbons (PAHs) and chlorinated PAHs (ClPAHs) are ubiquitous contaminants that bind to the aryl hydrocarbon receptor (AhR) and exhibit mutagenic potential. It is difficult to monitor human exposure levels to ClPAHs because the exposure routes are complicated, and environmental concentrations are not always correlated with the levels of PAHs. Urinary PAH metabolites are useful biomarkers for evaluating PAH exposure, and ClPAH metabolites may therefore contribute to the estimation of ClPAH exposure. One of the most abundant ClPAHs present in the environment is 1-chloropyrene (ClPyr), and urinary ClPyr metabolites have the potential to be good biomarkers to evaluate the level of exposure to ClPAHs. Since the metabolic pathways involving ClPAHs are still undetermined, we investigated the effect of human cytochrome P450 enzymes on ClPyr and identified three oxidative metabolites by liquid chromatography-tandem mass spectrometry and nuclear magnetic resonance. We found that ClPyr was metabolized most efficiently by the P450 1A1 enzyme, followed by the 1B1 and 1A2 enzymes. Similar to ClPyr, these metabolites were shown to have agonist activity for the human AhR. We detected these metabolites when ClPyr reacted with a pooled human liver S9 fraction as well as in human urine samples. These results suggest that the metabolites may be used as biomarkers to evaluate the extent of exposure to ClPAHs.

Benzotriazole ultraviolet stabilizers show potent activities as human aryl hydrocarbon receptor ligands.

Benzotriazole ultraviolet stabilizers (BUVSs) used in consumer products are raising concerns as new pollutants in the aquatic environment. We determined the agonistic activities of eight BUVSs and a chemically distinct UV absorber (4-methylbenzylidinecamphor) toward the human aryl hydrocarbon receptor (AhR) and thyroid hormone receptors alpha and beta. Although none of the BUVSs showed ligand activity against the thyroid hormone receptors, four of them (UV-P, UV-9, UV-326, and UV-090) showed significant AhR ligand activity. Their half-maximal effective concentrations (EC50) were 130 nM for UV-P, 460 nM for UV-9, and 5.1 µM for UV-090 (a value for UV-326 could not be determined). Of the numerous AhR ligands, it is well-known that those considered nontoxic are quickly metabolized by enzymes such as CYP1A1, which destroys their ability to function as ligands. Accordingly, we established a new yeast assay for simultaneous monitoring of both the strength of AhR ligand activity and ligand degradation by CYP1A1. We found the AhR ligand activities of the above four BUVSs to be stable in the presence of CYP1A1; therefore, they have the potential to accumulate and exert potent physiological effects in humans, analogous to polycyclic aromatic hydrocarbons and dioxins, which are known stable and toxic ligands.

Inhalation and dietary exposure to Dechlorane Plus and polybrominated diphenyl ethers in Osaka, Japan.

This study estimated daily exposure to Dechlorane Plus (DP) and polybrominated diphenyl ethers (PBDE) via inhalation and diet. Samples of atmospheric particles and food (obtained by market basket method) from Osaka, Japan were analyzed for DP (syn-, anti-) and PBDE using gas chromatography-mass spectrometry. DP was detected in both atmospheric particles and food samples. Among the atmospheric particles, DP was detected in all samples. ΣDP concentration was 7.1-15.4 pg m(-3) and anti-DP was the dominant residue among DP isomers. PBDE was also detected in all the atmospheric particles. ΣPBDE concentration was 9.9-23.3 pg m(-3). In the market basket study, DP was detected in Groups Ш (sugar and confectionary), V (legumes and their products), X (fish, shellfish, and their products), and XI (meat and eggs) at concentrations of 3.3, 2.8, 1.9, and 1.5 pg g(-1) wet wt, respectively. PBDE was detected in Groups Ш, IV (oils and fats), V, X, XI, and XШ (seasonings and other processed foods) at concentrations of 153, 79.1, 74.6, 308, 94.8, and 186 pg g(-1) wet wt, respectively. The daily intake of ΣDP (750 pg day(-1)) via inhalation and diet was approximately one percent of that for ΣPBDE (62 ng day(-1)).

Time trend of hexabromocyclododecane in the breast milk of Japanese women.

Hexabromocyclododecane (HBCD) is a type of brominated flame retardant. We investigated the time trend of the stereoisomer-specific concentrations of HBCD (alpha, beta, and gamma) in breast milk samples of Japanese women by using liquid chromatography-tandem mass spectrometry (LC/MS/MS). The pooled milk lipid samples collected in 1973, 1978, 1983, 1988, 1993, 1999, and from 2000 to 2006 were analysed. The concentration was below the detection limit in the samples collected between 1973 and 1983 and increased in those collected since 1988. Between 1988 and 2006, alpha-HBCD was detected in all 11 breast milk samples collected from 25 to 29-year-old Japanese women; gamma-HBCD, in 7; and beta-HBCD, in none of the samples. Time trend of summation operatorHBCD appeared to be related to that of the industrial HBCD consumption level in Japan.

Determination of the human cytochrome P450 monooxygenase catalyzing the enantioselective oxidation of 2,2',3,5',6-pentachlorobiphenyl (PCB 95) and 2,2',3,4,4',5',6-heptachlorobiphenyl (PCB 183).

2,2',3,5',6-Pentachlorobiphenyl (PCB 95) and 2,2',3,4,4',5',6-heptachlorobiphenyl (PCB 183) possess axial chirality and form the aS and aR enantiomers. The enantiomers of these congeners have been reported to accumulate in the human body enantioselectively via unknown mechanisms. In this study, we determined the cytochrome P450 (CYP) monooxygenase responsible for the enantioselective oxidization of PCB 95 and PCB 183, using a recombinant human CYP monooxygenase. We evaluated 13 CYP monooxygenases, namely CYP1A1, CYP1A2, CYP1B1, CYP2A6, CYP2B6, CYP2C8, CYP2C19, CYP2E1, CYP2J2, CYP3A4, CYP3A5, CYP4F2, and aromatase (CYP19), and revealed that CYP2A6 preferably oxidizes aS-PCB 95 enantioselectively; however, it did not oxidize PCB 183. The enantiomer composition was elevated from 0.5 (racemate) to 0.54. In addition, following incubation with CYP2A6, the enantiomer fraction (EF) of PCB 95 demonstrated a time-dependent increase.

Trends in the enantiomeric composition of polychlorinated biphenyl atropisomers in human breast milk.

For the precise estimation of the risk to human health caused by persistent organic pollutants (POPs), it is important to discuss enantiomer fraction value (EF value) because it is reported that behaviors such as stability and toxicity of enantiomers are quite different in human body. Among POPs, polychlorinated biphenyl (PCB) is known as one of the most persistent compounds in human breast milk samples. The main exposure source of PCB for human body is mostly from food especially in seafood. The contamination of fish and shellfish has been a serious problem for the Japanese, who consume a large amount of fish in their diet. PCBs have 19 congeners which are chlorine-substituted in 3- or 4- ortho positions are known to have enantiomers. In this study, we analyzed PCB 183 (2,2',3,4,4',5',6-hepta CB) in human breast milk and fish samples enantioselectively and revealed the time trends of the EF value. Though EF value of PCB 183 in fish samples sustained close to racemate (EF = 0.5) from 1982 to 2012, that in breast milk increased over time. This fact indicates that (+)-PCB-183 has greater bioaccumulation potential than (-)-PCB-183 in human body; therefore, the toxicity of (+)-PCB-183 should be emphasized.

Monitoring of Antibiotic Residues in Aquatic Products in Urban and Rural Areas of Vietnam.

Antibiotic residues in aquatic products in Vietnam were investigated. A total of 511 fish and shrimp samples were collected from markets in Ho Chi Minh City (HCMC), Thai Binh (TB), and Nha Trang (NT) from July 2013 to October 2015. The samples were extracted with 2% formic acid in acetonitrile and washed with dispersive C18 sorbent. Thirty-two antibiotics were analyzed by LC-MS/MS. Of the 362 samples from HCMC, antibiotic residues were found in 53 samples. Enrofloxacin was commonly detected, at a rate of 10.8%. In contrast, samples from TB and NT were less contaminated: only 1 of 118 analyzed samples showed residues in TB and only 1 of 31 showed residues in NT. These differences were attributed to the local manufacturing/distribution systems. To understand the current status of antibiotic use and prevent adverse effects that may be caused by their overuse, continual monitoring is required.

[Congener-specific analysis of Pcbs in food samples by using GC/MS].

A GC/MS method for congener-specific analysis of polychlorinated biphenyls (PCBs) in food samples was evaluated. There was good agreement between total concentration of PCBs in fish samples (n = 16) measured by using the GC/MS method and the conventional GC-ECD method. The total diet study samples (fishery food, meat, egg, milk, and their products) collected from Osaka in 1982-2001 were analyzed by using the GC/MS method to estimate the temporal trend of daily intake of PCBs from food. The estimated daily intake of total PCBs (sum of tri- to heptaCBs) was in the range of 0.7-4.4 microg/person/day, far below the Japanese provisional acceptable daily intake (250 microg/50 kg-person/day). The dominant congener was 2,2',4,4',5,5'-hexachlorobiphenyl (#153), which accounted for 9-15% of total PCB. The ratio of lower chlorinated congeners decreased from the 1980's to post-2000. The results indicate that the PCB congener profile in foods has gradually changed.

Rapid and easy multiresidue method for the analysis of antibiotics in meats by ultrahigh-performance liquid chromatography-tandem mass spectrometry.

This study involved the development of a multiresidue method for the rapid analysis of 43 antibiotics in meats using ultrahigh-performance liquid chromatography-tandem mass spectrometry. This method was performed using dispersive-solid phase extraction, which is able to analyze 20 samples within 2 h. All compounds were determined simultaneously on a C18 separation column with gradient elution. Validation of the analytical method was performed by carrying out linearity, limit of quantification (LOQ), accuracy, precision, and recovery tests in different meat products. The validation criteria were set according to AOAC International and Japanese validation guidelines. The linearity of each compound was almost the coefficient of determination (r(2)) > 0.98. The LOQs of all tested antibiotics were <10 µg/kg. The results verify that this method is capable of quantitative analysis of 36, 33, and 37 compounds in beef, pork, and chicken, respectively. This method can be used for rapid and easy multiresidue screening of antibiotics for three meats (pork, beef, and chicken).

Dechlorane Plus and decabromodiphenyl ether in atmospheric particles of northeast Asian cities.

Atmospheric particles were collected in several cities in Japan (Sapporo, Sagamihara, Kanazawa, and Kitakyushu), Korea (Busan), and China (Beijing) using a high-volume air sampler equipped with a quartz fiber filter. The summer and winter samples were analyzed using gas chromatography-high-resolution mass spectrometry for Dechlorane Plus (DP). Decabromodiphenyl ether (BDE-209) was also analyzed for the samples from Kanazawa and Beijing. DP was detected in all samples. The mean total DP (ΣDP) concentration was highest (6.7 pg/m(3)) and lowest (0.87 pg/m(3)) in the winter samples from Sagamihara and Busan, respectively. The seasonal variation of DP concentrations varied by sampling site in this study. BDE-209 was detected in all the analyzed samples except for one of the Kanazawa winter samples. BDE-209 concentration was considerably higher in Beijing than in Kanazawa. Significant correlations were found between the concentrations of ΣDP and BDE-209 in the winter samples from Kanazawa and in both summer and winter samples from Beijing. This similarity in the atmospheric behavior of DP and BDE-209, especially in winter, is assumed to reflect a common end usage and release mechanism.

Antibiotic residue monitoring results for pork, chicken, and beef samples in Vietnam in 2012-2013.

A monitoring plan of residual antibiotics in food of animal origin was conducted in Vietnam from 2012 to 2013. Meat samples were collected from slaughterhouses and retail stores in Ho Chi Minh City and Nha Trang. A total of 28 antibiotics were analyzed using a LC-MS/MS screening method. Sulfonamides, fluoroquinolones, and tilmicosin were detected in some of the samples. Sulfaclozine and fluoroquinolones were mainly detected in chicken samples, and sulfamethazine was mainly detected in pork samples. High levels of sulfonamide residues, ranging between 2500 and 2700 µg/kg sulfaclozine and between 1300 and 3600 µg/kg sulfamethazine, were present in two chicken and three pork samples, respectively. Tilmicosin was detected at ranges of 150-450 µg/kg in 10 chicken samples. Positive percentages were 17.3, 8.8, and 7.4% for chicken, pork, and beef, respectively, for an average of 11.9%. The results suggest an appropriate withdrawal period after drug administration had not been observed in some livestock.

Atmospheric chlorinated polycyclic aromatic hydrocarbons in East Asia.

This study estimates atmospheric concentrations of chlorinated polycyclic aromatic hydrocarbons (ClPAHs) and polycyclic aromatic hydrocarbons (PAHs) in East Asia using a Gas Chromatograph with High Resolution Mass Spectrometer (GC-HRMS). ClPAHs are ubiquitously generated from PAHs through substitution, and some ClPAHs show higher aryl hydrocarbon receptor (AhR)-mediated activities than their parent PAHs. Atmospheric particles were collected using a high-volume air sampler equipped with a quartz-fiber filter. We determined the ClPAH concentrations of atmospheric particles collected in Japan (Sapporo, Sagamihara, Kanazawa, and Kitakyushu), Korea (Busan), and China (Beijing). The concentrations of ClPAHs were highest in the winter Beijing sample, where the total mean concentration was approximately 15-70 times higher than in the winter samples from Japan and Korea. The concentrations of Σ19ClPAHs and Σ9PAHs were significantly correlated in the Kanazawa and the Busan samples. This indicates that within those cities ClPAHs and PAHs share the same origin, implying direct chlorination of parent PAHs. Toxic equivalent concentrations (TEQs) of the total ClPAHs and PAHs were lowest in Kanazawa in the summer, reaching 1.18 and 2610fg-TEQm(-3) respectively, and highest in Beijing in the winter, reaching 627 and 4240000fg-TEQm(-3) respectively.

Detection of Dechlorane Plus and brominated flame retardants in marketed fish in Japan.

Fish samples purchased from Japanese markets were analyzed for Dechlorane Plus (DP)(syn-, anti-), polybrominated diphenyl ether (PBDE), and hexabromocyclododecane (HBCD)(α, γ). Twenty fish were analyzed using gas chromatography-mass spectrometry for DP and PBDE, and using liquid chromatography-tandem mass spectrometry for HBCD. DP was detected in 18 samples and ∑DP concentrations were <0.2-14.2 pg g(-1)wet wt. Among the DP isomers, anti-DP was the dominant residue observed in this study. PBDE was detected in all samples. Concentrations of ∑PBDE ranged from 2.2 to 878 pg g(-1) wet wt. HBCD was detected in 18 samples, and ∑HBCD concentrations were <0.02-21.9 ng g(-1)wet wt. In fish landed near the East China Sea and the Sea of Japan, we detected relatively high concentrations of DP, PBDE, and HBCD. These results indicate that the seawaters around East Asia are contaminated with flame retardants. This study demonstrates the presence of DP in fish marketed in Japan for the first time.

[Determination of melamine in Chinese-made processed food].

A method using liquid chromatograph coupled with tandem mass spectrometer (LC/MS/MS) was developed for the determination of melamine in processed food. After extraction with 50% acetonitrile and clean-up with PSA and SCX, the quantification limit of melamine in processed food was 0.5 microg/g. The recoveries and relative standard deviations were 113 to 115% and less than 5%, respectively. Therefore, we considered that the developed method offers high precision and sensitivity for the determination of melamine in processed food. When the method was applied to six suspected products, 0.8 to 37 microg/g of melamine was detected from four milk-rich products.

Surveillance of polychlorinated biphenyl congeneric patterns in human breast milk from 1973 to 2000 in Osaka, Japan.

OBJECTIVE: Because the analysis of polychlorinated biphenyls (PCBs) in 1970's was performed using a packed column gas chromatograph, with an electron capture detector to determine total-PCB concentration, analytical data obtained by this analysis do not meet the requirement for the risk assessment of PCB congeners. In this context, the present study was carried out to reevaluate the congeneric analysis data by analyzing breast milk fat specimens that have been kept frozen. METHODS: PCB congeners in human breast milk were analyzed by high-resolution gas chromatography/high-resolution mass spectrometry (HRGC/HRMS) using a capillary column for the selected ion monitoring (SIM) of PCBs. RESULTS: Twelve major PCB congeners were detected in breast milk. The concentrations of all congeners of PCBs in breast milk were found to decrease annually, and their changes over time differed greatly for each congener. Between 1973 and 2000, almost no changes in the proportions of the 12 congeners of heptachlorinated biphenyls (HpCBs) and hexachlorinated biphenyls (HxCBs) were found whereas a marked and a slight decrease in the proportion of pentachlorinated biphenyls (PeCBs) and tetrachlorinated biphenyls (TeCBs), respectively, were observed. CONCLUSIONS: The PCB contamination of the Japanese population is estimated to be largely caused by seafood intake, but the proportions of PCB congeners in fish commodities were different from those in breast milk. The absorption, metabolism and therefore the accumulation of PCBs in the human body differ greatly depending on the congener.