Dietary Exposure of the Japanese General Population to Elements: Total Diet Study 2013-2018.

Some countries have conducted a total diet study (TDS) focused on the estimation of specific trace elements. Although some results of a Japanese TDS examining trace elements were published, there have been no reports of a nationwide TDS across Japan over a multi-year period to estimate the level of exposure to multiple elements. In the present study, a TDS using a market basket approach was performed to estimate the dietary exposure levels of the general population of Japan to 15 elements, including aluminum (Al), total arsenic (tAs), boron (B), barium (Ba), cadmium (Cd), cobalt (Co), chromium (Cr), total mercury (THg), molybdenum (Mo), nickel (Ni), lead (Pb), antimony (Sb), selenium (Se), tin (Sn), and uranium (U). Samples prepared in eight regions across Japan over a 6-year period were analyzed using validated methods. The robust mean exposure estimates for Al, tAs, B, Ba, Cd, Co, Cr, THg, Mo, Ni, Pb, Sb, Se, Sn, and U were 48, 4.2, 29, 8.6, 0.35, 0.17, 0.49, 0.14, 4.2, 2.8, 0.15, 0.022, 1.8, 0.10, and 0.021 µg/kg body weight/day, respectively. Although the variability in exposure estimates varied greatly from element to element, the relative standard deviations calculated from the robust means and robust standard deviations were ≤ 50% for all elements except Sn. Compared against the health-based guidance values, none of the robust and precise estimates obtained for the target elements would be associated with urgent health risk concern. In addition, the estimated exposure levels were generally in agreement with previously reported estimates, indicating that health risks associated with exposure to these elements have not changed markedly nationwide in Japan in recent years.

Estimation of dietary intake of 90Sr in Japan after the Fukushima Daiichi Nuclear Power Plant accident: market basket study, 2013-2018.

Radionuclide contamination in foods has been a public concern in Japan after the Fukushima Daiichi Nuclear Power Plant (FDNPP) accident. To estimate time and regional trends of daily intake and annual committed effective dose of strontium-90 (90Sr) after the accident, we analysed Market basket samples using a low background 2π gas-flow counter. Samples were collected from six regions, once a year from 2013 to 2018. There appeared to be little variation in estimated daily intake and annual committed effective dose of 90Sr across the time periods and regions. The estimated maximum annual committed effective dose of 90Sr was 0.00076 mSv/year, a value sufficiently lower than the intervention exemption level, 1 mSv/year, in foods in Japan. There was no noticeable difference between the range of estimated daily intake of 90Sr in this study compared with daily intake measured before the FDNPP accident. These results suggested that no obvious increase in dietary intake of 90Sr was observed after the FDNPP accident, and that the effects on commercial foods from 90Sr due to the FDNPP accident were negligible.

Probabilistic Estimation of Dietary Intake of Methylmercury from Fish in Japan.

Dietary intake of methylmercury from fish was estimated via Monte Carlo simulation using data for methylmercury concentrations in 210 fish samples and data regarding fish consumption extracted from the Japanese National Health and Nutrition Survey. The fish analyzed were classified into 5 groups according to categories used in the survey. The distribution of consumption of fish from each group was used without fitting to statistical distributions. A log-normal distribution was fitted to the distribution of methylmercury concentration in each fish group. Two random numbers that followed these distributions were generated, and a trial value was calculated by multiplying these random numbers. The trial value was divided by the body weight (50 kg) to arrive at an estimate of dietary methylmercury intake. A total of 100,000 Monte Carlo simulation iterations were performed. The estimated mean daily intake of methylmercury was 0.093 µg/kg body weight (bw)/day. This value is well below the tolerable daily intake of 0.292 µg/kg bw/day calculated from the tolerable weekly intake (2.0 µg/kg bw/week) established by the Food Safety Commission of Japan. The probability that the daily intake of methylmercury exceeds the tolerable daily intake was 7.6%. As there are no data regarding fish consumption for consecutive days, estimation of the weekly intake of methylmercury is a subject for future studies.

[Validation Procedure for a Multi-Element Analytical Method Aiming at Dietary Intake Estimation].

Total diet study (TDS) is a useful way to estimate the dietary intake of hazardous and chemical substances. Regarding the analysis performed in TDS, international guidelines published by the World Health Organization, recommend selecting and confirming the validity of suitable analytical methods to achieve the purpose of TDS. However, concrete procedures and/or approaches for confirming the validity of suitable methods have yet to be established. In the present study, we aimed to develop samples, referred to as SEMPs; Samples to estimate methods performance, that can be used to evaluate the performance of the analytical methods applied to the composite samples prepared in TDS. The concentrations of 14 kinds of elements, including hazardous substances such as arsenic, lead, and cadmium, in SEMPs were measured for use in the validation of a multi-element analytical method for estimating dietary intake. After examining the appropriate amount of relevant elements added to the samples, we established a performance evaluation procedure by repeatedly analyzing five fortified and non-fortified SEMPs each.

Continuous Estimation of Annual Committed Effective Dose of Radioactive Cesium by Market Basket Study in Japan from 2013 to 2019 after Fukushima Daiichi Nuclear Power Plant Accident.

Radionuclide contamination in foods has been a great concern after the Fukushima Daiichi Nuclear Power Plant (FDNPP) accident. To estimate time trends of daily intake and annual committed effective dose of radionuclides after the accident, radioactive cesium (r-Cs; 134Cs and 137Cs) and potassium-40 (40K) in market basket (MB) samples prepared at 6-month intervals in periods from September 2013 to March 2019 in 15 regions of Japan were analyzed using γ-ray spectrometry. The annual committed effective dose of r-Cs, calculated at non-detected radionuclide levels assumed to be half the limit of detection (LOD), appeared to decrease gradually in 11 regions close to the FDNPP that were more likely to be affected by the accident. Differences in doses among the 15 regions were large just after the accident, but gradually decreased. In particular, 134Cs has not been detected in any MB sample in any region since September 2018, and annual committed effective dose from 134Cs in all regions was mostly constant at around 0.3 µSv/year (given the respective LODs). The maximum annual committed effective dose of r-Cs in this study was decreased from 2.7 µSv/year in September 2013 to 1.0 µSv/year in March 2019. In contrast, the range of annual committed effective dose of 40K varied from approximately 150 to 200 µSv/year during that time frame and did not change much throughout the period of this study. Although annual committed effective doses of r-Cs in regions close to the FDNPP appeared to be higher than in regions far from the FDNPP, doses in all regions are remaining at a much lower levels than the intervention exemption level, 1 mSv/year, in foods in Japan.

Concentrations of Polycyclic Aromatic Hydrocarbons in Smoked Foods in Japan.

ABSTRACT: We evaluated the performance of a gas chromatography-tandem mass spectrometry method for quantifying 16 polycyclic aromatic hydrocarbons (PAHs), which the Scientific Committee on Food and the Joint Food and Agriculture Organization of the United Nations/World Health Organization Expert Committee on Food Additives have considered to be of concern to human health and used the method to determine the PAH concentrations in smoked foods. Eighty-seven samples of smoked fish, smoked meat, smoked eggs, dried bonito flakes, and dried bonito-related soup-stock products (disposable powder packets for infusion, instant bouillons, and liquids) were purchased in Japan to analyze their content of the 16 PAHs. Because of the low certainty of some results, the analytical values for some PAHs (e.g., benzo[c]fluorene, chrysene, and dibenzo[a,h]pyrene) are given for informational purposes only. The highest median concentrations of benzo[a]pyrene and the sum of all the 16 PAHs (29 and 760 µg/kg, respectively) were found in the disposable powder packets, followed by dried bonito flakes (24 and 512 µg/kg, respectively), and instant bouillons (11 and 227 µg/kg, respectively). These concentrations were much higher than those in the other products tested. We also investigated the percentages of the PAHs transferred from dried bonito flakes and a disposable powder packet to soup stocks commonly prepared at home. These were extremely low (<4%), even though they contained relatively high concentrations of the PAHs. Finally, the intake of benzo[a]pyrene and the sum of the intakes of four PAHs, as a marker proposed by the European Food Safety Authority, were estimated based on the data from a Japanese food consumption survey and the mean concentrations found in smoked fish and smoked fish products. These estimates suggest intakes of PAHs pose a low concern for consumer health.

[Development of a Rapid Determination Method for Strontium-90 in Foods Using a Strontium Extraction Column for Use during Emergency Events].

To ensure food safety during emergency events such as nuclear disasters, we developed a practical rapid determination method for strontium-90 (Sr-90) in foods. Purification of Sr from foods was simplified using a commercial Sr-extraction column. We also reduced the waiting time to achieve radiative equilibrium between Sr-90 and Y-90. Finally, we developed a rapid determination method for Sr-90 that can be completed in about a week. Using the new method, stable Sr recoveries exceeded 85%. The trueness of the method ranged from 109 to 115% and the detection limit of Sr-90 was estimated to be 0.07 Bq/kg fresh weight according to a performance evaluation using standard materials. Sr-90 radioactivity concentrations in food samples determined by the new method were highly correlated and nearly equal to concentrations determined by the conventional method. The present study suggests that the new method offers highly sensitive and rapid detection of Sr-90 which are necessary attributes for food tests during emergency events.

[Concentration of Radioactive Cesium in Domestic Foods Collected from the Japanese Market in Fiscal Years 2014-2016].

Between fiscal years 2014 and 2016, we surveyed the concentration of radioactive cesium in commercial foods produced in areas where there is a risk of radiation contamination due to the Fukushima Daiichi nuclear disaster. The number of samples with a concentration of radioactive cesium that exceeded the regulatory limit （100 Bq/kg for general foods） was 9 out of 1,516 （0.6％） in fiscal 2014, 12 out of 900 （1.3％） in fiscal 2015, and 10 out of 654 （1.5％） in fiscal 2016. Even though some samples were expected to be contaminated with radioactive cesium, because wild mushrooms and edible wild plants were intentionally included in this survey, the percentage of samples that exceeded the regulatory limit was only around 1％. The surveillance results confirmed that the pre-shipment food monitoring conducted by local governments was properly and efficiently performed, although continuous monitoring of the concentration of radioactive cesium in cultivated and wild mushrooms, edible wild plants, and wild animal meats is still required.

Dietary intake of dioxins in Japan in 2016 with time trends since 1998.

Total diet samples collected from seven regions throughout Japan in 2016 were analysed for polychlorinated dibenzo-p-dioxins, polychlorinated dibenzofurans and dioxin-like polychlorinated biphenyls (DL-PCBs), known collectively as dioxins. This led to estimates of the latest dietary intake of these contaminants for the general Japanese population (≥1 year old). The average daily intake of dioxins for a person weighing 50 kg, calculated at non-detected congener concentrations assumed to be equal to zero, was estimated to be 0.54 pg TEQ (toxic equivalents) kg-1 body weight (bw) day-1. This value is well below the tolerable daily intake of 4 pg TEQ kg-1 bw day-1 for dioxins in Japan. The average intake was highest from fish and shellfish, followed by meat and eggs. The TEQ contribution of the fish and shellfish group to the total dietary TEQs was significant (89%). The DL-PCBs accounted for about 67% of the dioxin intake. The latest dioxin intake level was compared with previous estimates from total diet study results obtained annually since 1998 to determine the time trends in the dietary intake of dioxins in Japan. Overall, the average dioxin intake appeared to be decreasing gradually during the period of study. The previous average intakes of dioxins ranged from 0.58 to 1.9 pg TEQ kg-1 bw day-1. The latest average intake was the lowest since 1998 and was about one-third of the average intake in 1998. This decreasing trend in the dietary intake of dioxins was mainly influenced by the decreased dioxin intakes from two food groups, fish and shellfish, and meat and eggs.

Dioxin concentrations in dietary supplements containing animal oil on the Japanese market between 2007 and 2014.

We determined the concentrations of dioxins (polychlorinated dibenzo-p-dioxins, dibenzofurans and dioxin-like polychlorinated biphenyls) in 46 dietary supplement products, containing the oil of fish, marine mammals, or egg yolk, on the Japanese market between 2007 and 2014. Dioxins were detected in 43 of the 46 products tested at concentrations from 0.00015 to 67 pg TEQ/g. The highest concentration of dioxins was found in a shark liver oil product which varied insignificantly in five batches collected over a two-year period. The dioxin intakes from these five batches reached 2.3-2.8 pg TEQ/kg bw/day, or 58%-70%, respectively, of the Japanese tolerable daily intake (TDI) of 4 pg TEQ/kg bw/day. However, the dioxin intakes from most of the other products tested were less than 5% of the TDI. Although rare, supplements based on animal oils may contain relatively high concentrations of dioxins, leading to a substantial increase in dioxin intakes.

Surveillance of Total Mercury and Methylmercury Concentrations in Retail Fish.

Most fish samples contain methylmercury, that the concentrations very greatly according to the fish species. To avoid the adverse health effects of methylmercury while retaining the benefits provided by fish consumption, it is important to select suitable fish species and to control the amount of the fish intake. We surveyed the concentrations of total mercury and methylmercury in 210 retail fish samples classified into 19 fish species by using validated analytical methods. The results of this survey were as follows. The total mercury and methylmercury concentrations were higher than 1 mg/kg in some samples of swordfish and bluefin tuna, which are large predatory fish species. In bluefin tuna and yellowtail, total mercury and methylmercury concentrations in farm-raised fish were lower than those in natural fish. There was a positive correlation between total mercury concentration and methylmercury concentration. Our results indicate that a cut-off value of 0.3 mg/kg total mercury in the screening of fish samples would increase the effectiveness of inspection.

Determination of polychlorinated biphenyls in marine fish obtained from tsunami-stricken areas of Japan.

We determined the polychlorinated biphenyl (PCB) congeners in 101 marine fish obtained from tsunami-stricken areas following the Great East Japan Earthquake in 2011. In particular, to determine the degree of PCB contamination in the fish, we investigated the concentration of total PCB (∑PCB) and the proportions of 209 individual PCB congeners by high-resolution gas chromatography/high-resolution mass spectrometry. The ∑PCB concentration was 1.7-33 ng/g in fat greenling (n = 29), 0.44-25 ng/g in flounder (n = 36), and 1.6-86 ng/g in mackerel (n = 36), all values being much lower than the provisional regulatory limit in Japan. In the congener analysis, tetra-, penta-, hexa-, and hepta-chlorinated PCB congeners dominated in all samples (comprising over 86% of the ∑PCB). The proportions of the chlorinated PCB congeners were similar to the contamination patterns derived from Kanechlor in the environment, implying that the marine fish were not contaminated with fresh PCBs.

Validation Study on a Method for Simultaneous Quantification of Elements in Mineral Water and Surveillance of Element Concentrations.

A validation study was performed on the Japanese official method to determine several elements (B, Cr, Mn, Cu, Zn, As, Se, Cd, Ba, Pb) in mineral water simultaneously using ICP-MS. The trueness, repeatability and reproducibility of the method were estimated to be in the ranges of 95-106, 0.2-1.4 and 0.4-4.2%, respectively. The values of every performance parameter fulfilled the requirements in the guidelines announced by the Ministry of Health, Labour, and Welfare of Japan, and the validity of the method was confirmed. The concentrations of the elements in mineral water products purchased from Japanese markets in 2013 and 2014 were surveyed using the method. All products met the specifications defined in the Japanese Food Sanitation Law. No clear relationship between the element concentrations and the survey year or the producing country was found.

Simultaneous determination of seven anticoagulant rodenticides in agricultural products by gel permeation chromatography and liquid chromatography-tandem mass spectrometry.

A sensitive and reliable method for the simultaneous determination of hydroxycoumarin-type (brodifacoum, bromadiolone, coumatetralyl, and warfarin) and indandione-type (chlorophacinone, diphacinone, and pindone) rodenticides in agricultural products by gel permeation chromatography (GPC) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) was developed. The procedure involved extraction of the rodenticides from samples with acetone, followed by liquid-liquid partitioning with hexane/ethyl acetate (1:1, v/v) and 10% sodium chloride aqueous solution, then cleanup using GPC, and finally, analysis using LC-MS/MS. High recoveries from the GPC column were obtained for all rodenticides tested using a mobile phase of acetone/cyclohexane/triethylamine (400:1600:1, v/v/v). An ODS column, which contains low levels of metal impurities, gave satisfactory peak shapes for both hydroxycoumarin- and indandione-type rodenticides in the LC-MS/MS separation. The average recoveries of rodenticides from eight agricultural foods (apple, eggplant, cabbage, orange, potato, tomato, brown rice, and soybean) fortified at 0.0005-0.001 mg/kg ranged from 76 to 116%, except for bromadiolone in orange (53%) and diphacinone in soybean (54%), and the relative standard deviations ranged from 1 to 16%. The proposed method effectively removed interfering components, such as pigments and lipids, and showed high selectivity. In addition, the matrix effects were negligible for most of the rodenticide/food combinations. The results suggest that the proposed method is reliable and suitable for determining hydroxycoumarin- and indandione-type rodenticides in agricultural products.

Detection of Aryl Hydrocarbon Receptor Activation by Some Chemicals in Food Using a Reporter Gene Assay.

The purpose of this study was to examine whether a simple bioassay used for the detection of dioxins (DXNs) could be applied to detect trace amounts of harmful DXN-like substances in food products. To identify substances with possible DXN-like activity, we assessed the ability of various compounds in the environment to bind the aryl hydrocarbon receptor (AhR) that binds specifically to DXNs. The compounds tested included 19 polycyclic aromatic hydrocarbons (PAHs), 20 PAH derivatives (nitrated, halogenated, and aminated derivatives), 23 pesticides, six amino acids, and eight amino acid metabolites. The AhR binding activities (AhR activity) of these compounds were measured using the chemical activated luciferase gene expression (CALUX) reporter gene assay system. The majority of the PAHs exhibited marked AhR activity that increased in a concentration-dependent manner. Furthermore, there was a positive link between AhR activity and the number of aromatic rings in the PAH derivatives. Conversely, there appeared to be a negative correlation between AhR activity and the number of chlorine residues present on halogenated PAH derivatives. However, there was no correlation between AhR activity and the number and position of substituents among nitrated and aminated derivatives. Among the pesticides tested, the indole-type compounds carbendazim and thiabendazole showed high levels of activity. Similarly, the indole compound tryptamine was the only amino acid metabolite to induce AhR activity. The results are useful in understanding the identification and characterization of AhR ligands in the CALUX assay.

Surveillance of Strontium-90 in Foods after the Fukushima Daiichi Nuclear Power Plant Accident.

As a result of the Fukushima Daiichi nuclear power plant (NPP) accident, various radionuclides were released into the environment. In this study, we surveyed strontium-90 ((90)Sr) concentrations in several foodstuffs. Strontium-90 is thought to be the third most important residual radionuclide in food collected after the Fukushima Daiichi, NPP accident after following cesium-137 ((137)Cs) and cesium-134 ((134)Cs). Results of (90)Sr analyses indicated that (90)Sr was detect in 25 of the 40 radioactive cesium (r-Cs) positive samples collected in areas around the Fukushima Daiichi NPP, ranging in distance from 50 to 250 km. R-Cs positive samples were defined as containing both (134)Cs and (137)Cs which are considered to be indicators of the after-effects of the Fukushima Daiichi NPP accident. We also detected (90)Sr in 8 of 13 r-Cs negative samples, in which (134)Cs was not detected. Strontium-90 concentrations in the r-Cs positive samples did not significantly exceed the (90)Sr concentrations in r-Cs negative samples or the (90)Sr concentration ranges in comparable food groups found in previous surveys before the Fukushima Daiichi NPP accident. Thus, (90)Sr concentrations in r-Cs positive samples were indistinguishable from the background (90)Sr concentrations arising from global fallout prior to the Fukushima accident, suggesting that no marked increase of (90)Sr concentrations has occurred in r-Cs positive samples as a result of the Fukushima Daiichi NPP accident.

Performance Evaluation of an Improved GC-MS Method to Quantify Methylmercury in Fish.

Here, we set out to improve our previously developed methylmercury analytical method, involving phenyl derivatization and gas chromatography-mass spectrometry (GC-MS). In the improved method, phenylation of methylmercury with sodium tetraphenylborate was carried out in a toluene/water two-phase system, instead of in water alone. The modification enabled derivatization at optimum pH, and the formation of by-products was dramatically reduced. In addition, adsorption of methyl phenyl mercury in the GC system was suppressed by co-injection of PEG200, enabling continuous analysis without loss of sensitivity. The performance of the improved analytical method was independently evaluated by three analysts using certified reference materials and methylmercury-spiked fresh fish samples. The present analytical method was validated as suitable for determination of compliance with the provisional regulation value for methylmercury in fish, set in the Food Sanitation haw.

[Development of ICP-OES, ICP-MS and GF-AAS Methods for Simultaneous Quantification of Lead, Total Arsenic and Cadmium in Soft Drinks].

In this study, we developed methods to quantify lead, total arsenic and cadmium contained in various kinds of soft drinks, and we evaluated their performance. The samples were digested by common methods to prepare solutions for measurement by ICP-OES, ICP-MS and graphite furnace atomic absorption spectrometry (GF-AAS). After digestion, internal standard was added to the digestion solutions for measurements by ICP-OES and ICP-MS. For measurement by GF-AAS, additional purification of the digestion solution was conducted by back-extraction of the three metals into nitric acid solution after extraction into an organic solvent with ammonium pyrrolidine dithiocarbamate. Performance of the developed methods were evaluated for eight kinds of soft drinks.

[Surveillance of radioactive cesium in domestic foods on the Japanese market (Fiscal Years 2012 and 2013)].

We surveyed the concentration of radioactive cesium in foods purchased at markets in areas where possible contamination has been a concern after the Fukushima accident. In fiscal years 2012 and 2013, we surveyed 1,735 and 1,674 foods, respectively, using a NaI (Tl) scintillation spectrometer for the screening test and a γ-ray spectrometer with a germanium semiconductor detector for the final test. Only 3 and 4 samples (0.2% of our total samples) exceeded the regulatory limit (100 Bq/kg) for radioactive cesium in fiscal years 2012 and 2013, respectively. Our surveillance indicates that the pre-shipment monitoring of foods by local governments has been working effectively.

Simultaneous determination of acidic pesticides in vegetables and fruits by liquid chromatography--tandem mass spectrometry.

A sensitive and efficient method has been developed for the simultaneous determination of 73 multi-class acidic pesticides, such as phenoxy acid and sulfonylurea herbicides, in vegetables and fruits. The sample preparation procedure was carefully optimized for the efficient removal of co-extracted matrix components. The method involves extraction of acidic pesticides with acetonitrile containing hydrochloric acid, removal of water from crude extract by salting out, and sequential cleanup by octadecylsilyl silica gel and silica gel columns. For samples containing high amounts of pigments, such as spinach, additional cleanup using a graphitized carbon column was performed prior to liquid chromatography-mass spectrometry (LC-MS/MS) analysis. Recovery tests were performed for five times for each sample of cabbage, spinach, potato, eggplant, orange, and apple fortified at 0.01 mg kg-1. Out of the 73 tested pesticides, 70 for cabbage, 67 for spinach, 69 for potato, 67 for eggplant, 64 for orange, and 70 for apple were within the range of 70-120%, with relative standard deviations below 25%. Nitenpyram and pyrasulfotole showed low recoveries for all the samples tested, probably due to low recoveries from silica gel column. The developed method effectively removed co-extracted matrix components and was highly selective, with no interfering peaks found in the chromatograms of blank samples. The overall results indicate that the developed method is suitable for the quantitative analysis of acidic pesticide residues in vegetables and fruits.