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As an analytical method for aflatoxins in foods, the analytical method based on the notification by the director of the Food Safety Department, Pharmaceutical and Food Safety Bureau, Ministry of Health, Labour and Welfare (August 16, 2011) has been established. In order to improve the operability and analytical performance of the conventional method, this study aimed to construct an improved method that optimized selection of immunoaffinity column (IAC) and purifying condition, and omitted evaporation after the purification with IAC. In the recovery test performed by adding 2.5 ng/g of aflatoxin B1, B2, G1 and G2 standard solutions into 9 kinds of food samples, the improved method achieved the established target values: 77.0-99.7% of recovery, 1.7-5.6% of intra-assay coefficient of validation, and 0.9-3.6% of inter-assay of coefficient of variation, respectively. The improved method also achieved 4.3-10.5% greater recovery and 1.5 hours shorter preparation time than the conventional one. These results indicate applicability of the improved method for 9 kinds of foods and its efficacy as an analytical method for aflatoxins in foods.

In this study, we developed an LC-MS/MS-based rapid and simple analytical method for six fungicides; imazalil, o-phenylphenol, thiabendazole, fludioxonil, azoxystrobin and pyrimethanil, the latter three were newly approved for use after 2011. For expediting and simplification, we merged the extraction method with that of the pesticide analysis. For purification step, loading of 1 mL of sample extracts to 500 mg Oasis HLB column and elution with 8 mL of acetonitrile gave satisfactory results. The performance of the present method was confirmed for orange, grapefruit, and lemon samples fortified with the six fungicides. The results showed that the average recovery ranged from 89.7 to 100.0%, intra- and inter-assay CV% ranged from 1.5 to 5.0% and from 0.5 to 4.9%, respectively, achieving the target values of the Japanese official guideline for residual pesticide analysis. The limits of quantification of this method were determined to be 1 mg/kg for o-phenylphenol, and 0.2 mg/kg for the other five fungicides. These values were lower than their corresponding regulation values. In addition, we confirmed the usability of the present method for fungicide inspection of commercially available citrus fruits. During 2017-2019, there was no conflict between the food labeling and the fungicides detected and no fungicide with the concentration exceeding maximum residue level was detected.

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 (240 ng/g lipid weight), followed by dichlorodiphenyltrichloroethane and its metabolites (DDTs) (150 ng/g lipid weight), chlordane-related compounds (CHLs) (36 ng/g lipid weight), hexachlorobenzene (HCB) (16 ng/g lipid weight), hexachlorocyclohexanes (HCHs) (4.4 ng/g lipid weight), polybrominated diphenyl ethers (PBDEs) (1.8 ng/g lipid weight), and finally, Mirex (1.5 ng/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).

The aim of this study was to develop a monoclonal antibody (mAb)-based enzyme-linked immunosorbent assay (ELISA) for the quantification of a major allergen (Cit s 2) in fresh and processed oranges. Purified recombinant Cit s 2 (rCit s 2)-small ubiquitin-like modifier (SUMO) was used for the production of mAbs. In the optimized ELISA, the recovery of rCit s 2 from Navel oranges or orange juice was 107–132%, and the intra- and inter-assay coefficients of variation were 3.1–8.8% and 4.4–11%, respectively. The Cit s 2 content in fresh oranges was determined to be 1,800 ± 430 ng/g, while this content was much lower in the processed foods. The developed ELISA demonstrated high reproducibility, sensitivity, and accuracy, and this assay may help individuals with orange allergy by determining Cit s 2 quantities in food products and controlling their Cit s 2 intake.

Purpose Prenatal exposure to environmental chemicals is a growing concern, because such exposures have been shown to be associated with various diseases. The levels of chemicals and heavy metals in maternal blood, cord blood, maternal urine and amniotic fluid in Japanese pregnant women were investigated. Methods A total of 145 women, including 14 fetal growth restriction cases, were included in the present study. The levels of phthalates (di[2-ethylhexyl]phthalate and mono[2-ethylhexyl]phthalate), perfluorinated compounds (perfluorooctane sulfonate, perfluorohexanoic acid, perfluorooctanoic acid, and perfluorononanoic acid), pesticides (dimethylphosphate, dimethylthiophosphate, diethylphosphate, diethylthiophosphate, 3-phenoxybenzoic acid, and octachlorodipropyl ether), bisphenol A, nicotine (nicotine, nornicotine, cotinine, norcotinine, and trans-3’-hydroxycotinine), polybrominated diphenyl ethers, and heavy metals were measured. The relationship between fetal growth and the levels of chemicals and heavy metals were investigated. Results Phthalates, perfluorinated compounds, pesticides, polybrominated diphenyl ethers, and heavy metals were detected in high frequency, whereas nicotine and bisphenol A were almost negative. Phthalates, perfluorinated compounds, and several heavy metals were transferred to the fetus. High perfluorononanoic acid levels in the maternal blood and cord blood, and low perfluorooctanoic acid level in the cord blood were significantly and negatively associated with fetal growth. Conclusions The present study showed that pregnant women in Japan and their fetuses are exposed to a variety of chemicals and heavy metals.

To detect the tomato allergen profilin (Sola l 1) from fresh tomato and tomato-containing processed foods, we generated a rabbit polyclonal anti-Sola l 1 peptide antibody. An indirect enzyme-linked immunosorbent assay (iELISA) based on the antibody was established for semiquantification of Sola l 1, and standard curves were generated using Sola l 1 homologue recombinant birch profilin (rBet v 2) as a calibrator. The mean concentration of tomato extracts was 25.6 ± 9.1 µg/g, and the concentrations of tomato-containing processed foods ranged from 13.3 to 24.8 µg/g. The reactivity of Sola l 1 after heat and papain treatment of tomato extracts was evaluated using competitive iELISA, and the results showed that Sola l 1 reactivity was stable with or without treatment. Our findings suggested that tomato-containing processed foods and fresh tomatoes both contained Sola l 1 and could have the potential to induce allergic symptoms in patients with tomato allergies.

We carried out a collaborative study in six laboratories to confirm the universality of the enhancing effect of co-existing reference pesticides on the GC-MS peak response to a target pesticide (malathion, procymidone, or flucythrinate). First, we confirmed the response enhancement of the target pesticides with increasing numbers of co-existing reference pesticides in solution. Then, using diluted green soybean matrix, we analyzed the target pesticides with two types of matrix-matched calibration, containing the target pesticides or 166 other pesticides. In both cases, the response-enhancing effect of co-existing pesticides was confirmed in all laboratories. The enhancement was reduced by addition of green soybean matrix to the sample and calibration solutions. Our results show that it is necessary to estimate the peak response-enhancing effect of co-existing pesticides in the calibration solution to obtain accurate results with GC-MS determination. The enhancing effect could be reduced by addition of food matrix to the sample and calibration solutions.

The total mercury (T-Hg) and methylmercury (MeHg) concentrations in red snow crabs (Chionoecetes japonicus) caught off the coast of Japan were analyzed. The T-Hg concentration ranged from 0.03 to 0.56 mg/kg (mean: 0.21 mg/kg) in the raw muscle, and 0.02 to 0.74 mg/kg (mean: 0.27 mg/kg) in the boiled muscle. The MeHg concentration ranged from 0.04 to 0.54 mg/kg (mean: 0.20 mg/kg) in the raw muscle. The mean ratio of MeHg to T-Hg was 0.88. The crab body weight was found to significantly correlate with the concentrations of T-Hg (r = 0.488) and MeHg (r = 0.490) (p ≤ 0.01). For the general population in Japan, the intake of MeHg from eating red snow crab was estimated to be lower than 0.013 mg/week, which was less than one-sixth of the tolerable MeHg intake (0.08 mg/week).

Oranges (Citrus sinensis) can provoke an allergic reaction in some individuals. The concentration of profilin (Cit s 2) – the major allergen in oranges – is unknown, but this knowledge could be useful for preventing the development of allergies. The aim of this study was to quantify the amount of Cit s 2 in Navel oranges and in different varieties of citrus fruit. A sandwich enzyme-linked immunosorbent assay (ELISA) was developed using both monoclonal and polyclonal antibodies. Extracts of Navel oranges and 12 citrus fruits were tested using this ELISA. Cit s 2 levels in the Navel orange pulp and peel were 1.81 ± 0.74 and 1.19 ± 0.87 µg/g, respectively, with accumulation detected primarily in the pulp. These findings may help in the production of low-risk processed oranges for orange allergy sufferers through selection of orange varieties and parts with low profilin content during food processing.

The number of students with food allergy is currently increasing. Moreover, the unintentional mixing or accidental ingestion of allergy-causing food materials in school lunches has attracted great attention. The aim of this study was to verify the current status of elementary school lunch provision for students with food allergy. We investigated the elementary school lunch services in seven cities in Osaka prefecture. The egg elimination diet was provided in five of the seven cities. In four of these five cities, we did not detect the presence of egg residue either on the surface of various cookware used to prepare the egg elimination diet or in the food itself. In this investigation, the egg elimination diet was provided properly, but we observed differences among the cities in the manual preparation of foods for food allergy diets. To step up these efforts, our results suggest the necessity of preparing a manual to consider individual conditions of school lunch services.

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 ).

In vitro fertilization (IVF) is an important treatment for infertility, and in recent years, the delivery rate in IVF has increased markedly with the progress of reproductive medicine. However, chemical contaminants in IVF media have not been fully studied. A simple method using highperformance liquid chromatography coupled with tandem mass spectrometry was therefore investigated for quantitation of bisphenol A (BPA), 4nonylphenol (NP) and 4-n-nonylphenol (n-NP) in IVF media. Media samples were processed using the solid-phase extraction method. The quantitation limits of BPA, NP, and n-NP were 0.5, 5 and 0.5 ng·ml-1, respectively, and the recoveries for BPA, NP and n-NP added to the media samples were above 90%. Commercially available media samples for IVF including 15 media for incubation of ova, 9 sperm washing media (SWM), 4 human serum albumin (HSA) samples and 2 substitute serum (SS) samples were examined, with 2 lots for each sample. NP was detected in 3 media for incubation of ova, 12 SWM, 7 HSA samples and 4 SS samples, in ranges of 6.3 14.5, 5.3 21.7, 5.5 164.0 and 9.0 19.0 ng·ml-1, respectively. BPA and n-NP were not detected in any of the samples.

In multiresidue pesticide analysis using gas chromatography, it has long been recognized that an increase in the number of pesticides present in a standard solution can result in an enhancement of the peak responses of certain pesticides. Despite being widely acknowledged, this phenomenon has been rarely studied and is poorly understood. In this study, the authors have tentatively called this phenomenon the "matrix-like effect" and demonstrated it clearly using gas chromatography with tandem mass spectrometry. Five selected pesticides, namely, omethoate, terbufos, malathion, procymidone, and permethrin, and four internal standard candidates, namely, triphenyl phosphate, naphthalene-d8 , phenanthrene-d10 , and fluoranthene-d10 , were used to evaluate the matrix-like effect following the addition of 58, 108, and 166 other pesticides. With the exception of naphthalene-d8 , the responses of all evaluated pesticides and internal standard candidates were dramatically enhanced by the addition of up to 166 coexisting pesticides. The relative response factors of the five pesticides to each internal standard candidate were not constant under the conditions studied, meaning that these internal standard candidates did not adequately compensate for the matrix-like effect, at least for the five evaluated pesticides. The results revealed that the presence of various mixtures of pesticides in standard solutions might act as an unintentional analyte protectant, that is, some sort of troublesome "quasi-matrix."

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

In vitro fertilization (IVF) is one of the most important treatments of infertility to provide a chance of conceiving. In IVF treatment, sperm are washed and motile sperm are isolated with sperm washing media (SWM) for the purpose of fertilization; fertilized ova are then incubated for a maximum of 5 or 6d in media for IVF (IVFM). The exposure of fertilized ova to chemicals via such media has not been studied. We determined the concentrations of two contaminants; di(2-ethylhexyl)phthalate (DEHP) and its hydrolyzed product mono(2-ethylhexyl)phthalate (MEHP) in IVFM, SWM, and protein sources (PS: human serum albumin or serum substitute) for IVFM and SWM. The DEHP and MEHP in these media were extracted by a liquid-liquid extraction method and their concentrations determined by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). Fifteen IVFM, nine SWM, and six PS obtained in Japan were examined. The concentrations of DEHP and MEHP in IVFM and SWM were

Epigenetic alteration is an emerging paradigm underlying the long-term effects of chemicals on gene functions. Various chemicals, including organophosphate insecticides and heavy metals, have been detected in the human fetal environment. Epigenetics by DNA methylation and histone modifications, through dynamic chromatin remodeling, is a mechanism for genome stability and gene functions. To investigate whether such environmental chemicals may cause epigenetic alterations, we studied the effects of selected chemicals on morphological changes in heterochromatin and DNA methylation status in mouse ES cells (ESCs). Twenty-five chemicals, including organophosphate insecticides, heavy metals and their metabolites, were assessed for their effect on the epigenetic status of mouse ESCs by monitoring heterochromatin stained with 4¢,6-diamino-2-phenylindole (DAPI). The cells were surveyed after 48 or 96 h of exposure to the chemicals at the serum concentrations of cord blood. The candidates for epigenetic mutagens were examined for the effect on DNA methylation at genic regions. Of the 25 chemicals, five chemicals (diethyl phosphate (DEP), mercury (Hg), cotinine, selenium (Se) and octachlorodipropyl ether (S-421)) caused alterations in nuclear staining, suggesting that they affected heterochromatin conditions. Hg and Se caused aberrant DNA methylation at gene loci. Furthermore, DEP at 0.1 ppb caused irreversible heterochromatin changes in ESCs, and DEP-, Hg- and S-421-exposed cells also exhibited impaired formation of the embryoid body (EB), which is an in vitro model for early embryos. We established a system for assessment of epigenetic mutagens. We identified environmental chemicals that could have effects on the human fetus epigenetic status.

This study presents the results of a total diet study performed for estimating the dietary intake of polybrominated diphenyl ethers (PBDEs) in Osaka, Japan. The concentrations of 36 PBDEs were measured in samples from 14 food groups (Groups I-XIV). PBDEs were detected only in Groups IV (oils and fats), V (legumes and their products), X (fish, shellfish, and their products), and XI (meat and eggs) at concentrations of 1.8, 0.03, 0.48, and 0.01 ng g⁻¹, respectively. For an average person, the lower bound dietary intakes of penta- and deca-formulations were estimated to be 46 and 21 ng day⁻¹, respectively. A high proportion of the decabrominated congener (DeBDE-209) was observed in Group IV. To confirm the presence of DeBDE-209 in vegetable oils, an additional analysis was performed using 18 vegetable oil samples. Of these, seven contained ng g⁻¹ levels of DeBDE-209.

We analyzed the dietary fish and marine mammal oil supplements purchased from Japanese markets for hexabromocyclododecane (HBCD); 20 brands were analyzed by liquid chromatography–tandem mass spectrometry (LC/MS/MS). HBCD was detected in 15 of 22 samples, and the concentrations of all the HBCD isomers (α, β, and γ) ranged from

Polybrominated diphenyl ethers (PBDEs) are widely used flame retardants; currently, they are identified as ubiquitous environmental contaminants. Several studies indicate that PBDEs might affect male fertility. We present the results of a pilot study on the relationship between human serum PBDEs and sperm quality. The PBDE levels in Japan are comparable to those found in European countries. Strong inverse correlations were observed between the serum concentration of 2,2′,4,4′,5,5′-hexabromodiphenyl ether and sperm concentration (r = -0.841, p = 0.002) and testis size (r = -0.764, p = 0.01). Extensive studies on the relationship between PBDEs and sperm quality are required. © 2008 Springer Science+Business Media, LLC.

Many methods for the determination of pesticide residues in food have been reported. Although natural medicines should be confirmed to be as safe as food, few methods for the determination of pesticide residues in natural medicines have been reported. In this study, 17 organochlorine pesticides were detected in natural medicines using GC/MS with negative chemical ionization (NCI). GC/MS with NCI can detect halogenated pesticides selectively and thus is suitable for the detection of organochlorine pesticides. This study indicates that GC/MS with NCI is useful for analyzing organochlorine pesticides in natural medicines.

フタル酸モノエステル類（PMEs）には，精巣毒性を有するものがあり，当毒性に感受性の高い胎児及び乳幼児への暴露が懸念されている．しかし，母乳中のPMEsに関する報告は少ない．そこで，高速液体クロマトグラフィー/タンデム型質量分析器を用いて母乳中のPMEs｛フタル酸モノエチル，フタル酸モノブチル，フタル酸モノベンジル，フタル酸モノ(2-エチルヘキシル)，フタル酸モノ(2-エチル-5-ヒドロキシヘキシル) 及びフタル酸イソノニル｝の分析法を構築した．試料に内部標準を添加後，酢酸エチル/シクロヘキサン混液で抽出し，得られた有機相を乾固した．酢酸アンモニウム緩衝液に再溶解後，脱抱合反応を施し，固相抽出法により精製して試験液とした．PMEs（5.0 ng/mL添加）の回収率は，96.3∼103% であった．本分析法を活用し，日本人の母乳について11検体を分析した結果，フタル酸モノ(2-エチル-5-ヒドロキシヘキシル) を除く5種類のPMEs を検出した．

The regulation of transcription and genome stability by epigenetic systems are crucial for the proper development of mammalian embryos. Chemicals that disturb epigenetic systems are termed epimutagens. We previously performed chemical screening that focused on heterochromatin formation and DNA methylation status in mouse embryonic stem cells and identified five epimutagens: diethyl phosphate (DEP), mercury (Hg), cotinine, selenium (Se), and octachlorodipropyl ether (S-421). Here, we used human induced pluripotent stem cells (hiPSCs) to confirm the effects of 20 chemicals, including the five epimutagens, detected at low concentrations in maternal peripheral and cord blood samples. Of note, these individual chemicals did not exhibit epimutagenic activity in hiPSCs. However, because the fetal environment contains various chemicals, we evaluated the effects of combined exposure to chemicals (DEP, Hg, cotinine, Se, and S-421) on hiPSCs. The combined exposure caused a decrease in the number of heterochromatin signals and aberrant DNA methylation status at multiple gene loci in hiPSCs. The combined exposure also affected embryoid body formation and neural differentiation from hiPSCs. Therefore, DEP, Hg, cotinine, Se, and S-421 were defined as an “epimutagen combination” that is effective at low concentrations as detected in maternal peripheral and cord blood.

A method was established for the determination of 180 pesticide residues in fruits and vegetables. The procedure involved extraction with acetonitrile, followed by a salting-out step with anhydrous MgSO4 and NaCl. Removal of sediment and water was performed simultaneously by centrifugation. Co-extractives were removed with a double-layered SPE column, and graphitized carbon black and primary secondary amine (GCB/PSA) solid phase extraction cleanup cartridge. The eluate was determined by GC/FPD and GC/MS without further cleanup. Recovery data were obtained by fortifying 9 matrices at 0.05–0.1 µg/g. Recoveries of 180 pesticides were mainly 70–110% and the relative standard deviation (RSD) was below 25%. Limits of detection ranged between 0.01 and 0.05 µg/g for tested pesticides. © Pesticide Science Society of Japan

The time-trend and recent concentrations of polybrominated diphenyl ethers (PBDEs) in Japanese mother's milk were investigated. The time-trend of 16 PBDEs (BDE-28, 37, 47, 66, 71, 75, 77, 85, 99, 100, 119, 153, 154, 138, 183, and 190) in pooled milk samples from mothers living in Osaka between 1973 and 2000 was analyzed. Additionally, PBDE concentrations in individual milk samples collected from 13 mothers living in Kanagawa and Okayama in 1999 were measured. The total concentration of all PBDE congeners (SigmaPBDEs) measured in the pooled samples increased during the period between 1973 (0.01 ng/g lipid) and 1988 (1.64 ng/g lipid), and remained low afterwards while showing remarkable changes in PBDE congener profiles. The SigmaPBDEs in the 1999 individual milk samples were also low (0.56-3.97 ng/g lipid), except for a single sample (291 ng/g lipid). The source of this exposure could not be identified.

In this study, airborne particulates were collected at three sites, two in a downtown area and the other in a suburban area of Kanazawa, Japan in each season for 7 years. Two polycyclic aromatic hydrocarbons (PAHs), pyrene (Py) and benzo[a]pyrene (BaP) and four nitropolycyclic aromatic hydrocarbons (NPAHs), 1-nitropyrene (NP) and 1,3-, 1,6-, and 1,8-dinitropyrenes (DNP) were determined by high-performance liquid chromatography with fluorescence and chemiluminescence detection. At the downtown sites, the mean concentration of each DNP was about two orders of magnitude lower than that of 1-NP and more than three orders of magnitude lower than those of Py and BaP. This tendency reflected the composition of PAHs and NPAHs in diesel-engine exhaust particulates. Concentrations of these PAHs and NPAHs were higher at the downtown sites than at the suburban site, suggesting the dilution of these compounds during the transportation from the downtown to the suburban area. The concentration ratios of NPAHs to PAHs were larger at the downtown sites than at the suburban site. Studies using UV light and sunlight showed that degradation of NPAHs was faster than that of PAHs. Thus, the lower concentrations of NPAHs in the suburban sites may be due to their being photodegraded faster than PAHs during the atmospheric transportation from the downtown area to the suburban area.

Airborne particulates were collected at a site near a group of factories in Kitakyushu, one of the most industrialized cities in Japan, and at downtown sites in Sapporo and Tokyo, typical large commercial cities. We determined the levels of polycyclic aromatic hydrocarbons (PAHs) and nitropolycyclic aromatic hydrocarbons (NPAHs), which are known to be carcinogenic and/or mutagenic. The atmospheric concentrations of most PAHs with 4-, 5-, and 6-rings were higher in Kitakyushu and Tokyo than in Sapporo. On the other hand, atmospheric concentrations of the strongly mutagenic NPAHs were the highest in Sapporo, intermediate in Tokyo, and lowest in Kitakyushu. The atmospheric concentrations of 1-nitropyrene (1-NP) in Kitakyushu were one order of magnitude lower than those in the two commercial cities. Therefore the concentration ratios of NPAHs to their nucleus PAHs were significantly smaller in Kitakyushu than in the two commercial cities. This result suggested that the atmospheric levels of NPAHs in Kitakyushu were comparatively low, although the air in Kitakyushu was as heavily polluted with PAHs as the air in Tokyo. The concentration ratio of dinitropyrenes (DNPs) to 1-NP in Kitakyushu was much higher than that in the other two cities. A possible reason for the above two differences between the two types of cities is the contribution of chimney exhaust of steel manufacturing plants in Kitakyushu, which contains high concentrations of PAHs.

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.

A high-throughput multiresidue analysis of pesticides in non-fatty vegetables and fruits was developed. The method consisted of a single extraction and a single clean-up procedure. Food samples were extracted with ethyl acetate and the mixture of extract and food dregs were poured directly into the clean-up column. The clean-up column consisted of two layers of water-absorbent polymer (upper) and graphitized carbon (lower), which were packed in a reservoir (75 ml ) of a cartridge column. The polymer removed water in the extract while the carbon performed clean-up. In a recovery test, 110 pesticides were spiked and average recoveries were more than 95% from spinach and orange. Most pesticides were recovered in the range 70–115% with RSD usually

Airborne particulates were collected during the same periods in downtown Kanazawa, Sapporo and Tokyo and polycyclic aromatic hydrocarbons (PAHs) and nitropolycyclic aromatic hydrocarbons (NPAHs) were determined together with several heavy metal elements and water-soluble inorganic ions. The mean concentrations of metal elements and inorganic ions were the highest in Tokyo, followed by Sapporo and the lowest in Kanazawa both in winter and summer, and were strongly dependent on the amount of airborne particulates. These tendencies were different from the cases of PAHs and NPAHs. Correlations between the organic compounds (PAHs and NPAHs) and several inorganic pollutants were strong only in winter samples of Sapporo, but not so strong in Kanazawa and Tokyo. Because PAHs and NPAHs and several combustion-origin inorganic substances are, in general, detected mainly in the fine airborne particulates, it was reasonable that the significant correlations were found only in the winter samples of Sapporo where the generation of the coarse and crustal particulates were suppressed by snow or ice. Though the atmospheric concentrations of PAHs and NPAHs in Kanazawa were much lower than in Sapporo, the correlation among the NPAH compounds and between PAHs and NPAHs were surprisingly similar in these two cities. The strong correlations indicated that the atmospheric behaviors of the PAHs and NPAHs studied were similar in the two cities and that the source of PAHs and NPAHs might be the same. On the contrary, strong correlations were not observed in Tokyo, suggesting the possibility of multiple sources or formations/degradations of PAHs and

Eight residual triazine herbicides and three metribuzin metabolites in foods were determined by liquid chromatography mass spectrometry (LC-MS) with an atmospheric pressure chemical ionization (APCI) interface, under both positive and negative ion modes. Herbicides were extracted with acetonitrile, and no cleanup procedure was adopted in this method. Four foods were spiked with eight herbicides and three metabolites at 0.05 ppm. The average recoveries of these herbicides usually ranged from 82 to 99% and the relative standard deviations were usually around 10%. These results suggest that LC-MS with APCI can be used to determine residues of triazine herbicides in foods.

A single extraction and a single clean-up procedure was developed for multi-residue analysis of pesticides in non-fatty vegetables and fruits. The method involves the use of a high capacity absorbent polymer for water as a drying agent in extraction from wet food samples and of a graphitized carbon column for clean-up. A homogeneously chopped food sample (20 g) and polymer (3 g) were mixed to absorb water from the sample and then 10 min later the mixture was vigorously extracted with ethyl acetate (100 ml). The extract (50 ml), separated by filtration, was loaded on a graphitized carbon column without concentration. Additional ethyl acetate (50 ml) was also eluted and both eluates were concentrated to 5 ml for analysis. The procedure for sample preparation was completed within 2 h. In a recovery test, 107 pesticides were spiked and average recoveries were more than 80% from asparagus, orange, potato and strawberry. Most pesticides were recovered in the range 70-120% with usually less than a 10% RSD for six experiments. The results indicated that a single extraction with ethyl acetate in the presence of polymer can be applied to the monitoring of pesticide residues in foods.

In May 2012, two schoolchildren developed allergic symptoms after eating a school meal of fried bread in Osaka, Japan. One specific raw material, milk, should not be present in fried bread. However, we suspected the unintentional mixing of milk in the bread manufacturing process. Our aim was to verify the reason if this was so.We first manufactured bread that contained milk as one of its components. We then thoroughly cleaned the manufacturing line except for the dough divider and, as per the revised instruction manual, continuously manufactured bread that did not contain milk. In this manner, we tried to simulate the conditions at the time of the incident. Casein is a major milk protein. We, therefore, determined casein concentration in the milk-free bread, using casein enzyme-linked immunosorbent assay.We observed casein levels higher than 1000 ppm in the initial lots of breads, but lower levels in the later lots. Casein levels also decreased upon frying of bread.High casein levels, which were observed in the earlier lots of breads, were lower in the later lots. In the manufacturing line, the remnants of the bread dough that contained milk presumably got mixed with the dough used in the subsequent manufacture of milk-free bread.

The concentrations of 10 polybrominated diphenyl ethers (PBDEs) in commercial culture media and protein sources (PSs) for in vitro fertilization (IVF) of human ova were investigated. Samples of 15 IVF media (IVFM), nine sperm preparation media (SPM), and six PSs were analyzed. PBDEs were detected in 10 IVFM, seven SPM, and all PS samples in ranges of 0.6-35, 0.9-31, and 7.5-385pgg(-1), respectively. A dominant PBDE congener BDE-47 was detected in the PS and PS-supplemented samples. Our findings suggested that PS supplementation was the potential cause of PBDE-contamination of IVFM and SPM.

Results are reported of a pilot survey of concentrations of polybrominated diphenyl ethers (PBDEs) and polychlorinated biphenyls (PCBs) in shark liver oil supplements. Eleven brands of dietary supplements were analysed using an isotope dilution GC/MS method. Total concentrations of 10 PBDE congeners (BDE-28, −47, −49, −66, −99, −100, −153, −154, −155 and −183) ranged from 0.1 to 53 ng g−1 oil weight and total concentrations of six PCB congeners (CB-28/31, −52, −118, −153 and −180) in the samples ranged from 16 to 340 ng g−1 oil weight (undetected values are not included). Two brands of Japanese deep-sea shark liver oil contained the highest levels of PBDEs (49–53 ng g−1 oil weight) and PCBs (290–340 ng g−1 oil weight). These results indicate that PBDEs may have entered Japanese deep-sea waters.

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.

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.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.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.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.

A rapid and simple extraction method for the simultaneous analysis of five neonicotinoid insecticides has been developed. Twelve different fruit and vegetable matrixes were extracted with methanol and cleaned up using a graphitized carbon solid phase extraction cartridge loading with a 20% methanol solution. The concentrated eluate after methanol elution was then analyzed for pesticide residues by liquid chromatography/mass spectrometry in the APCI positive mode. The five pesticides including nitenpyram, thiamethoxam, imidacloprid, acetamiprid, and thiacloprid were recovered at 70-95% at spike levels of 0.1 and 1 mg/kg in bell pepper, cucumber, eggplant, grape, grapefruit, Japanese radish, peach, pear, potato, rice, and tomato. Relative standard deviations were less than 10% for all of the recovery tests. The proposed method is fast, easy to perform, and could be utilized for regular monitoring of pesticide residues.

Residues of 17 pesticides in agricultural products were determined by LC/MS with an atmospheric pressure chemical ionization (APCI) interface in both positive and negative ion modes. Pesticides were extracted with acetonitrile, and the extracts were cleaned-up with a primary and secondary amine (PSA) mini-column eluted with acetone-hexane (1:1). Rice, orange and potato were spiked with the 17 pesticides at 0.1 microgram/g and analyzed by the proposed method. The average recoveries of these pesticides usually ranged from 70 to 98% and the relative standard deviations were usually around 10%. These results suggested that LC/MS with APCI could be used to determine the residue levels of the 17 pesticides in these crops.

Determination of 3 neonicotinoid insecticides, nitenpyram, imidacloprid, and acetamiprid, was studied. Vegetables and fruits were extracted with acetonitrile. The crude extract was passed through a weak anion-exchange cartridge (PSA). The effluent was subjected to silica gel cartridge. Imidacloprid and acetamiprid were eluted with 10 mL of 4:6 (v/v) acetone/hexane, followed by nitenpyram with acetone (20 mL). Pesticides were determined by HPLC with a C-18 column and diode-array detection system. Imidacloprid and acetamiprid were recovered at about 90% at the spike levels with 0.2 and 2 mg/kg in cucumber, potato, tomato, eggplant, Japanese radish, and grape. Nitenpyram was recovered at 64-80%. Relative standard deviations were less than 10% throughout all the recovery tests. In the residue analysis, agriculturally incurred pesticides at 0.08-0.14 mg/kg were designated with UV spectra compared with respective reference standards.

Development of an analytical method for polybrominated diphenyl ethers (PBDEs) in fish and their concentration in Japanese marine fish were investigated. Fish homogenate was extracted with diethyl ether/hexane (1+3). The extract was cleaned up by automated gel permeation chromatography (GPC) and then by mini-column chromatography, which consisted of three layers of silica gel and sulfuric acid-impregnated silica gel. The PBDE fraction was concentrated and injected into a GC/MS with negative chemical ionization (NCI). Recoveries of the 15 individual PBDEs (BDE-15, 28, 37, 47, 66, 71, 75, 77, 85, 99, 100, 119, 153, 154, and 209) each at a fortification level of 4 ng/g lipid were in the range of 88–128% and the relative standard deviations (RSD) were 0.43–7.6% ( n = 4). Seven species of marine fish (conger eel, flounder, gray mullet, horse mackerel, red sea bream, sea bass, and yellowtail) were collected from the Inland Sea of Seto, and were analyzed with the developed method. Seven PBDEs (BDE-28, 47, 66, 99, 100, 153, and 154) were detected in all the samples. The most abundant PBDE congener was BDE-47 found in all the samples. Relatively high levels of PBDEs were found in the gray mullets and yellowtails.

Water concentration in organic solvents after liquid-liquid partition was determined by the Karl Fischer titration method. n-Hexane and petroleum ether showed quite low levels of water, such as 0.1 mg/mL. The water concentration in wet ethyl acetate was about 20-30 mg/mL and that in diethyl ether was about 8-10 mg/mL. Anhydrous sodium sulfate absorbed about 20-25% of the water after vigorous mixing with wet ethyl acetate or diethyl ether. Wet acetonitrile extract from wet food, which contained about 60 mg/mL water after salting out with sodium chloride, was not dried at all with anhyfrous sodium sulfate treatment. Spiking n-hexane into wet ethyl acetate or wet diethyl ether was effective to exclude water. Spiking toluene into salted acetonitrile drove out water and dissolved sodium chloride. It can be concluded that the drying ability of anhydrous sodium sulfate towards wet organic solvents is poor, but it is effective in removing suspended water in solvents.

Various metal powders were batch screened as possible on-line reducers for nitropolycyclic aromatic hydrocarbons (NPAHs), in particular 1,3-dinitropyrene (1,3-DNP), 1,6-DNP, 1,8-DNP and 1-nitropyrene (1-NP) and its metabolites. Zinc was found to be an excellent reducer in chemiluminescent compatible mobile phases without any associated disposal problems. Zinc reducer column parameters, namely pH, buffer concentration, packing composition (zinc:glass beads or zinc:silica gel) and column length were all optimized for the HPLC determination system with chemiluminescence detection, named NPAH analyser. The system consisted of three pumps, sample injector, reducer column packed with zinc:glass beads, switching valve with a cutting loop, separation column (ODS), chemiluminescence detector and integrator. The mobile phase for the separation column and chemiluminescence reagent solution were respectively acetonitrile:imidazole–HClO4 and acetonitrile containing bis(2,4,6-trichlorophenyl)oxalate and H2O2. When the reducer column was introduced before the separation column, 1,3-, 1,6-, 1,8-DNPs and 1-NP in benzene:ethanol extracts from airborne particulates were separately determined with detection limits of 2–50 fmol, respectively, in a 100 μL sample. When the reducer column was introduced just after the separation column, 1-NP, 1-nitrosopyrene and 1-aminopyrene in the incubation mixture of 1-NP and the Salmonella typhimurium YG1021 strain were separately determined with detection limits as low as sub fmol levels. 1,8-DNP and its metabolite were also separately determined in the incubation mixture of 1,8-DNP and the S. typhimurium TA98 strain.

Polybrominated diphenyl ethers (PBDEs) have been used worldwide as additive flame retardants in polymeric materials. Commercial products consist predominantly of deca-, octa-, and pentabromodiphenyl ether mixtures. PBDEs are resistant to degradation in the natural environment and Penta-BDE in particular accumulates in the fatty tissues of fish, birds and mammals (including humans). Several toxic effects on the thyroid system or on neurodevelopment have been reported in experimental animals exposed to PBDEs. It is likely that human exposure is predominantly through the ingestion of contaminated food and/or mother's milk. The potential health effects of dietary exposure to PBDEs have now become a great concern because of the increasing PBDE levels in the biosphere. In this review, published information on the toxicology of PBDEs, levels in foodstuffs and human milk, and analytical methods has been compiled.

Air samples were collected at the sides of busy traffic roads in downtown Kanazawa, Sapporo and Tokyo during the same periods both in the daytime and in the night, in winter and summer. We determined the levels of polycyclic aromatic hydrocarbons (PAHs) and nitropolycyclic aromatic hydrocarbons (NPAHs), which are known to be carcinogenic and/or mutagenic. PAHs and NPAHs were analyzed simultaneously with an HPLC system with fluorescence detection for PAHs and another HPLC system with chemiluminescence detection for NPAHs, respectively. The amounts of airborne particulates were larger in the daytime than in the night in all three cities. This result was thought to reflect the diurnal change of such urban activities as traffic volume. Many of the PAH and NPAH compounds showed higher concentrations in the daytime than in the night, and also showed higher concentrations in winter than in summer in the three cities. This result coincided with the trends observed in our continuous surveys in downtown Kanazawa for several years. However, 5-ring PAHs, such as benzo [a] pyrene, and dinitropyrenes in Tokyo did not show such diurnal or seasonal differences. The composition ratio of NPAHs and the contribution of diesel-engine vehicles to the atmospheric concentrations of NPAHs were calculated from all the data of this survey. The concentration ratio of dinitropyrenes to 1-nitropyrene became larger with an increase in the ratio of diesel-engine vehicles to the total vehicles registered, suggesting a large contribution of diesel-engine vehicles to the concentrations of these NPAHs in urban air.

Polycyclic aromatic hydrocarbons (PAH) and nitropolycyclic aromatic hydrocarbons (NPAH) in airborne particulates collected simultaneously at downtown and suburban sites and in exhaust particulates from diesel and gasoline engine vehicles were determined. The mean concentrations of all compounds in the atmosphere were lower at the suburban site. The differences between the two sites were smaller in airborne particulates than in the atmosphere. However, the difference in NPAH concentrations in particulates between the two sites was greater than the difference in PAH concentrations, suggesting that the NPAH were less stable than the PAH. The concentrations of 1-nitropyrene and 1,3-, 1,6- and 1,8-dinitropyrenes were generally much higher in automobile exhaust particulates than in airborne particulates because the former were collected directly. However, 2-nitrofluoranthene and 2-nitropyrene were observed in airborne particulates but not in automobile exhaust particulates. The diurnal concentrations s...