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In order to avoid side-effects of lithium doses in some patients, some commonly cultivated mushroom species including A. bisporus have been successfully lithiated, with the potential to provide more acceptable sources of Li. This study assessed the in vitro release (potential bio-accessibility) and possible intake of Li using the action of artificial gastrointestinal juices on lithiated and non-lithiated (control) button mushrooms (Agaricus bisporus) that were subjected to certain modes of culinary processing.In the in vitro release study, mushrooms were processed using a number of routinely used domestic treatments including re-hydrating dried mushrooms, blanching, and blanching followed by pickling of fresh or frozen mushrooms. The in vitro digestion procedure used artificial gastrointestinal juices in a two-stage methodology that was adapted from "The Bioaccessibility Research Group of Europe" method. The Li concentrations were determined using an inductively coupled argon plasma - dynamic reactive cell - mass spectrometer.Lithium was found to be more bio-accessible from caps of lithiated mushrooms compared to non-lithiated. Releases from the caps and stipes of blanched or blanched and then pickled mushrooms, through gastric digestion ranged from 32 ± 2 to 50 ± 1% relative to the dried product and was lower for gastrointestinal digestion which ranged from 16 ± 1 to 20 ± 1%.Losses of Li sustained through blanching or blanching followed by pickling of fresh mushrooms (41 to 87% wet weight) combined with limited accessibility during gastrointestinal release (16 to 55%) result in much lower bio-availability of the dose from lithiated products. A 300 g meal would provide less than 5% of the Li (6 mg) required for potential preventative treatments, such as reducing suicide rates and lowering dementia risk.

The properties of potential emerging persistent contaminants, perfluoroalkyl substances (PFAS), in an andosol rice paddy lysimeter were analyzed to determine their mobility and leaching behavior regarding carbon chain length and functional groups. For this purpose, simulated contaminated water (ΣPFAS = 1,185,719 ng/L) was used in the lysimeter. The results showed that PFAS distribution in the paddy soil lysimeter was influenced by the migration of these substances into irrigation water and their adsorption into the soil. PFHxS (C6) and PFOS (C8), which are the main components of the simulated contaminated water, were mostly captured in the soil layers of the low-humic andosol layer (0–35 cm). PFAS distribution may depend on soil properties, such as total carbon (TC) content. Compared with perfluoroalkane sulfonic acids (PFSAs), the distribution of perfluoroalkyl carboxylic acids (PFCAs) in soil showed significant variation. The remaining PFCAs were distributed across all layers of the lysimeter, except for the longer-chain PFCAs. Moreover, the PFSA distribution was directly correlated with the carbon chain number, whereby longer- and shorter-chain PFSAs accumulated in the top and bottom soil layers, respectively. This study provides detailed information on the distribution, leaching, uptake, and accumulation of individual PFAS in andosol paddy fields in Japan.

Information regarding the size-dependent distribution of per- and polyfluoroalkyl substances (PFAS) in atmospheric particulate matter (PM) is very limited. In this study, 248 size-specific PM samples were collected from 9 Asian cities using a portable 4-stage cascade impactor for the analysis of PFAS. Of the 34 investigated PFAS, perfluorooctanoic acid (PFOA) and perfluorooctanesulfonic acid (PFOS) were the major compounds. In particular, the emerging PFAS, hexafluoropropylene oxide dimer acid, was quantified in the PM for the first time, with concentrations ranging from

Given that only a small number of per- and polyfluoroalkyl substances (PFAS) are routinely monitored, levels of PFAS in the atmosphere may be underestimated. A protocol including analyses of target PFAS (n = 50), water-soluble fluoride, and total fluorine has been proposed and applied to atmospheric samples. The whole method recovery (including extraction recovery and sampling efficiency) of 90-110% were obtained for the majority of compounds (48/50) with low deviations between replicates (20%). Fluorotelomer alcohols were the most prevalent PFAS in the indoor air, while the outdoor air was dominated by the ultrashort-chain ionic PFAS (e.g., trifluoroacetic acid and perfluoropropanoic acid). Concentrations of organofluorine (OF) compounds calculated from the fluorine mass balance ranged from 1.74 ng F/m

Perfluoroalkyl substances (PFASs) are a group of contaminants of concern in agricultural crops, but little is known of their accumulation or behavior in grains. We grew Japanese rice (Oryza sativa subsp. indica) in lysimeters irrigated with tap water or tap water plus simulated contaminated water for 2 years, then analyzed the roots, straw, unhulled rice, white rice, bran, soil, and water for PFASs residues. Total fluorine was measured by combustion ion chromatography. Estimated per-plant residue levels were 3.0 pg perfluorooctanesulfonic acid (PFOS) (bran: 0.5%, hull: 99.5%), 0.54 pg N-ethylperfluorooctanesulfonamide (N-EtFOSA) (white rice: 67%, hull: 33%), 1.2 pg perfluorobutanoic acid (PFBA) (white rice: 13%, bran: 7%, hull: 79%), 0.68 pg perfluoropentanoic acid (hull: 100%), 0.50 pg perfluorohexanoic acid (PFHxA) (white rice: 65%, bran: 16%, hull: 19%), 0.21 pg perfluoroheptanoic acid (hull: 100%), 0.25 pg perfluorooctanoic acid (PFOA) (hull: 100%), and 0.12 pg perfluorodecanoic acid (PFNA) (white rice: 81%, bran: 19%). Estimated daily PFASs intakes were1-3 ng perfluorooctanesulfonamide,1-7 ng N-EtFOSA, 1-2 ng PFBA,3-4 ng PFHxA, and 1-2 ng PFNA. Estimated PFOS, PFOA, and total PFASs in straw feed were 0.4, 0.1, and 2 kg yr

Hydrophobic pollutants, such as polychlorinated biphenyls (PCBs), are detected at high concentrations in plants of the Cucurbitaceae family, including cucumber and zucchini. Given that the uptake of these compounds by crops can lead to their accumulation in animals via the food chain and that they may have potentially adverse health effects, it is desirable to take appropriate preventative measures. However, the mechanisms underlying the uptake of these pollutants are still unclear. We found that the Magda (MG) cultivar of Cucurbita pepo showed high accumulation of PCBs, especially 2-chlorinated PCBs, in leaves, stems, and xylem sap compared with the Patty Green (PG) cultivar. In the MG cultivar, we identified two major latex-like proteins (MLPs) produced in roots that are translocated to xylem sap, wherein they bind to PCBs, and it is assumed that these MLPs are associated with the accumulation of PCBs in the aerial parts of plants. These results indicate that PCBs are transported by specific mechanisms via the xylem sap. Collectively, our findings indicate that the expression of MLP genes and production of MLPs in roots, the excretion of MLPs into xylem sap, and the binding of hydrophobic pollutants by MLPs are critical steps in the accumulation of hydrophobic pollutants in cucurbits.

An inter-laboratory trial (ILT) has been performed to validate ISO 21675 method for the measurement of per-and polyfluoroalkyl substances (PFAS) in water samples using solid phase extraction method and high-performance liquid chromatography-tandem mass spectrometry. A total of twenty-seven laboratories from eleven countries (Belgium: 1, Canada: 2, China: 2, France: 1, Germany: 3, Italy: 2, Japan: 6, Netherlands: 2, South Korea: 1, Sweden: 4, and USA: 3) participated in the ILT. Results of the homogeneity of ILT water samples showed that the repeatability tended to increase from short-chain to long-chain of PFAS. Results of stability of PFAS in Milli-Q water stored at 5 ± 3 °C ranged from 75% to 121% including those ultra-short-chain compounds, except for N-MeFOSA (44%), N-EtFOSA (44%), and 8:2 FTOH (30%) at 168 days. As for stability of PFAS in environmental waters, they were in acceptable range (between 70 and 125%) for most of PFAS, except for 8:2 FTUCA in the river water, seawater, and wastewater, and 8:2 FTSA and 8:2 FTOH in wastewater. Based on the performance data (reproducibility (CVR)

Loss of seed shattering has been regarded as a key step during crop domestication. Mutagenesis contributes to the development of novel crop cultivars with a desired seed-shattering habit in a relatively short period of time, but also to uncovering the genetic architecture of seed shattering. ‘Minamiyutaka’, a non-shattering indica rice cultivar, was developed from the easy-shattering cultivar ‘Moretsu’ by mutation breeding via gamma-ray irradiation. In present study, we observed significant differences in shattering habit, breaking tensile strength, and abscission zone structure between ‘Moretsu’ and ‘Minamiyutaka’. Whole-genome mutation analysis of ‘Minamiyutaka’ newly identified a 13-bp deletion causing defective splicing in exon 3 of the OsSh1 gene which has previously been referred to as a candidate for controlling seed shattering. Using CRISPR/Cas9 gene editing, we demonstrated that loss-of-function mutation in OsSh1 causes non-shattering in rice. Furthermore, gene expression analysis suggests that OsSh1 may function downstream of qSH1, a known key gene involved in abscission zone differentiation. Nucleotide diversity analysis of OsSh1 in wild rice accessions and cultivars revealed that OsSh1 has been under strong selection during rice domestication, and a missense mutation might have contributed to the reduction of seed shattering from the wild progenitors to cultivated rice.

Two sediment core samples from the brackish Lake Nakaumi in Japan were analyzed to determine the historical profiles of organochlorine pesticides (OCPs). It was observed that from the 1940s to 2005, the vertical distribution of OCPs in sediment cores reflected the temporal trend of pesticide usage in Japan. Dichlorodiphenyltrichloroethane (DDT) and its metabolites were predominant, with concentrations of 0.008-8.27 ng g

A new method is preliminarily validated for the simultaneous analysis of ionic and neutral per- and polyfluoroalkyl substances (PFASs) in both particulate and gaseous phases in air using a nanosampler-20 air sampler (NS20) composed of quartz fiber filters (QFFs), polyurethane foam (PUF) and artificial activated charcoal (GAIAC™). Perfluoroalkane sulfonamido ethanols (FOSEs) mainly remained in PUF, whereas the other neutral analytes were mainly found in GAIAC. Satisfactory recoveries were obtained for FOSEs, fluorotelomer alcohols (FTOHs), fluorotelomer iodides (FTIs), ranging fron 70%–120%, moderate recoveries were achieved for perfluorinated iodine alkanes (FIAs) and diiodofluoroalkanes (FDIAs), ranging from 50%–70%, while poor recoveries were found for perfluoroalkane sulfonamides (FOSAs). Breakthrough experiments revealed that almost all the target analytes were well trapped in GAIAC™, including the very volatile 4:2 FTOH. Applying to real sampling, our results showed that 6:2 and 8:2 FTOH were the most abundant species, with levels detected at 190 pg/m3 and 160 pg/m3. To the best of our knowledge, FDIAs were detected in ambient air for the first time at an average level of 8.3 pg/m3. Overall, the profiles observed from the real air samples reflected current industrial transition from longer chain to shorter chain in PFAS production. Our results revealed that the current method is promising for a more comprehensive understanding on the fates of PFASs in air.

Perfluoroalkyl substances (PFASs) were investigated in three types of vegetables (fruit, leafy, and root vegetables) that were cultivated and harvested from 2014 to 2017. The cultivated soil was mainly affected by perfluoroalkyl carboxylic acid (PFCAs; 91.8% detection rate) rather than perfluoroalkyl sulfonic acids (PFSAs; 8.2%). The cultivated soil (i.e., a volcanic cohesive soil) had a high total organic carbon (TOC = 3.4%) and therefore showed strong adsorption of long-chain PFASs. Short-chain PFCAs (i.e., under C9) were mainly detected in vegetables; specifically, PFBA showed high concentration in tomato shoots. Principal component analysis (PCA) plots clearly showed that PFASs in vegetables were different from those of cultivated soil, air, and rainwater. Interestingly, the whole potato (i.e., including peel) was in the same group as soil, indicating that the whole potato can easily be affected by the cultivated soil. Energy Dispersive X-ray Spectrometry-Scanning Electron Microscope (EDS-SEM) results showed that presence of unremovable micron-sized cultivated soil particles on the potato surface. Comparing the regional differences between the cultivated area of Tsukuba city (East Japan) and Osaka city (West Japan), PFASs patterns were similar in cucumber but differed in green perilla and potato.

Some cultivars of cucumbers, melons, pumpkins, and zucchini, which are members of the Cucurbitaceae family, are uniquely subject to contamination by hydrophobic pollutants such as the organohalogen insecticides DDT. However, the molecular mechanisms for the accumulation of these pollutants in cucurbits have not been determined. Here, cDNA subtraction analysis of Cucurbita pepo cultivars that are low and high accumulators of hydrophobic contaminants revealed that a gene for zinc finger proteins (ZFPs) are preferentially expressed in high accumulators. The cloned CpZFP genes were classified into 2 types: (1) the PBG type, which were expressed in C. pepo cultivars Patty Green, Black Beauty, and Gold Rush, and (2) the BG type, which were expressed in Black Beauty and Gold Rush. Expression of these CpZFP genes in transgenic tobacco plants carrying an aryl hydrocarbon receptor-based inducible gene expression system significantly induced β-glucuronidase activity when the plants were treated with a polychlorinated biphenyl (PCB) compound, indicating that highly hydrophobic PCBs accumulated in the plants. In transgenic tobacco plants carrying CpZFPs, accumulation of dioxins and dioxin-like compounds increased in their aerial parts when they were cultivated in the dioxin-contaminated soil. In summary, we propose that addition of CpZFP genes is a promising tool for conferring noncucurbits with the ability to accumulate hydrophobic contaminants.

An environmentally friendly water-based extraction method combined with conventional high-performance liquid chromatography with diode array detection is reported for the determination of seven hydrophilic neonicotinoid insecticides (dinotefuran, nitenpyram, thiamethoxam, clothianidin, imidacloprid, acetamiprid, and thiacloprid) in two fruiting vegetables (cucumber and eggplant). Insecticides were extracted from the vegetables with water and were preconcentrated with two solid-phase extraction cartridges packed with polymeric adsorbent, graphitized carbon black, and aminopropyl-modified silica gel. Matrix interferences were investigated. In order to accurately determine nitenpyram in cucumber and all neonicotinoids in eggplant, a matrix-matched calibration method was required. Average recovery values from the fortified samples were between 82% and 114%, with relative standard deviations less than 10%. The limits of quantification were 0.007–0.017 µg g−1 in cucumber and 0.003–0.019 µg g−1 in eggplant. The ...

Arsenic species, including arsenous acid, arsenic acid, methylarsonic acid, and dimethylarsinic acid, were determined using HPLC-ICPMS. The species were separated with a Discovery HS F5 column and a simple, volatile, and isocratic mobile phase of 0.1% (v/v) formic acid and 1% (v/v) methanol. The Discovery HS F5 column with a pentafluorophenyl (PFP) stationary phase gave sharp peaks and full separation of the arsenic species in 5min, and other PFP columns showed lower performance. This separation method was applied to arsenic species analysis in rice. The extraction of arsenic from rice samples was performed using 0.15M nitric acid. The methodology was validated by use of certified reference materials, NMIJ CRM 7503-a and NIST SRM 1568a, and extremely low arsenic rice samples as blank samples.

This is the first report, to our knowledge, to reveal important factors by which members of the Cucurbitaceae family, such as cucumber (Cucumis sativus), watermelon (Citrullus lanatus), melon (Cucumis melo), pumpkin (Cucurbita pepo), squash (C. pepo), and zucchini (C. pepo), are selectively polluted with highly toxic hydrophobic contaminants, including organochlorine insecticides and dioxins. Xylem sap of C. pepo ssp. pepo, which is a high accumulator of hydrophobic compounds, solubilized the hydrophobic compound pyrene into the aqueous phase via some protein(s). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of xylem sap of two C. pepo subspecies revealed that the amount of 17-kD proteins in C. pepo ssp. pepo was larger than that in C. pepo ssp. ovifera, a low accumulator, suggesting that these proteins may be related to the translocation of hydrophobic compounds. The protein bands at 17 kD contained major latex-like proteins (MLPs), and the corresponding genes MLP-PG1, MLP-GR1, and MLP-GR3 were cloned from the C. pepo cultivars Patty Green and Gold Rush. Expression of the MLP-GR3 gene in C. pepo cultivars was positively correlated with the band intensity of 17-kD proteins and bioconcentration factors toward dioxins and dioxin-like compounds. Recombinant MLP-GR3 bound polychlorinated biphenyls immobilized on magnetic beads, whereas recombinant MLP-PG1 and MLP-GR1 did not. These results indicate that the high expression of MLP-GR3 in C. pepo ssp. pepo plants and the existence of MLP-GR3 in their xylem sap are related to the efficient translocation of hydrophobic contaminants. These findings should be useful for decreasing the contamination of fruit of the Cucurbitaceae family as well as the phytoremediation of hydrophobic contaminants.

The transgenic tobacco plant XD4V-26 carrying the recombinant mouse aryl hydrocarbon receptor XD4V-mediated β-glucuronidase (GUS) reporter gene expression system was used for assay of dioxins and dioxin-like compounds consisting of polychlorinated dibenzeno-p-dioxins, polychlorinated dibenzofurans, and coplanar polychlorinated biphenyls (Co-PCBs) in actually contaminated soils. The transgenic tobacco plant XD4V-26 showed a significant dose-dependent induced GUS activity when cultured on MS medium containing PCB126 [toxic equivalency factor (TEF) = 0.1]. In contrast, PCB169 and PCB180, which have 0.03 of TEF and unassigned TEF values, respectively, did not significantly induce GUS activity under the same conditions as with PCB126. When the tobacco plants were cultivated for up to 5 weeks on actually contaminated soils with dioxins and dioxin-like compounds collected from the periphery of an incinerator used for disposal of residential and industrial wastes, GUS activity in the leaves was dose-dependently increased. The plants clearly detected 360 pg-TEQ g(-1) of dioxins and dioxin-like compounds in this assay. There was a positive correlation between GUS activity and TEQ value of dioxins and dioxin-like compounds in the plants. This assay does not require any extraction and purification processes for the actually contaminated soil samples.

Nine zucchini cultivars (Cucurbita pepo subspecies ovifera and pepo) were grown in soil containing a mixture of dioxins and dioxin-like compounds. Plants of subspecies pepo accumulated polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), and polychlorinated biphenyls (PCBs) in their aerial parts at higher concentrations than did plants of subspecies ovifera. In all cultivars, the accumulation of PCDDs and PCDFs in the aerial parts decreased with increasing hydrophobicity, whereas for PCBs, a negative correlation was observed in only two cultivars. The other seven cultivars selectively accumulated ortho-chlorinated biphenyls. Our results contribute to the understanding of the mechanisms underlying these phenomena, which should help in the development of efficient methods for phytoremediation of hydrophobic contaminants.

The concentrations of 12 POPs listed in the Stockholm convention, chlorophenols (CPs) and PAHs were investigated in sludge samples from wastewater and sewage treatment plants (WWTPs and STPs). The concentrations of PCDD/Fs in the wastewater sludge ranged from 0.189 to 1092 ng-TEQkg(-1) dry wt., and most of the sludge samples had levels below the EU guideline for the land application of PCDD/Fs (

Persistent organic pollutants, particularly polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), coplanar polychlorinated biphenyls (co-PCBs) and toxic equivalent quantity (pg TEQ g(-1)) were determined in sediment samples collected from Ariake Bay, Japan. The total concentration of PCDDs/DFs in surface sediment of the Yabe River (17,000 pg g(-1) dry wt) was approximately 2.5-fold higher than that of the Chikugo River (7,000 pg g(-1) dry wt) which has the highest flow amount of about 40% inputting to the northern Bay. The congener profile suggested that dioxin sources could be close to the study area. The PCDDs concentration (12,000 pg g(-1) dry wt) in the sediment layer, which was corresponding to the 1960s, was reflected on the pentachlorophenol (PCP) contamination occurring at that time in Ariake Bay. The reflecting of PCP in the sediment core was significantly considered by hierarchical cluster analysis. Based on isomer-specific analysis, the tetrachlorodibenzo-p-dioxin composition showed a gradual increase from 1978-1982 toward the surface, indicating that the pollution source at that period might be more influenced by chloronitrofen than PCP. This fact was supported, based on the ratio of Sigma PCDD to Sigma PCDF concentration. The PCDDs/DFs-derived TEQ contributed more than 90% of the SigmaTEQ (PCDDs/DFs and co-PCBs) in all the detected sediment layers. The contribution of PCBs to the total TEQ was low; however, an increasing historical trend of concentration was clearly observed.

An ELISA was used to directly determine residual imidacloprid in fruit juices. Imidacloprid could be determined by only diluting samples without any pre-treatments such as filtration, centrifugation, and clean-up procedures. The ELISA enabled imidacloprid to accurately determine down to about 5 μg/L in apple and grape juice samples and down to about 20 μg/L in orange juice sample. Recovery and precision of the ELISA were evaluated by spiking fruit juice samples with imidacloprid in the 10-400 μg/L ranges. Coefficients of variation were lower than 20% in all cases, and average recoveries were 94.2%, 113.2%, and 104.2% for apple, grape, and orange juice samples, respectively. No false positive results were found. The results obtained with the proposed ELISA well correlated with the reference HPLC for each fruit juice sample (r > 0.99).

The objective of this study was to clarify the adsorption and desorption characteristics of several herbicides in sediment. Five herbicides, esprocarb, thiobencarb, dimethametryn, pretilachlor, and simetryn were examined in this study. The adsorption ratio on the sediment increased in the following order: pretilachlor < dimethametryn < simetryn < thiobencarb < esprocarb. On the other hand, the adsorption ratio on the sediment without organic matter increased in the following order: thiobencarb < esprocarb < pretilachlor < dimethametryn < simetryn. Furthermore, the amounts of simetryn, dimethametryn, and pretilachlor adsorbed on the sediment without organic matter increased, while those of esprocarb and thiobencarb decreased in comparison to the original sediment. These results strongly suggested that the mineral surface in the sediment was very important as the adsorption site for the herbicide, especially in the case of simetryn, dimethametryn, and pretilachlor. All the adsorption and desorption data fitted well with the Freundlich equation. The hysteresis in the adsorption-desorption phenomena in the sediment was observed for all the herbicides, and it was affected by the organic matter in sediment, especially in the case of dimethametryn and pretilachlor.

The aquatic crustacean Daphnia magna is an important species for ecotoxicological study, and is often used as a test organism for environmental risk assessment. However, the mechanism of xenobiotic metabolism by this species has not been studied in detail. In the present study, pyrene was used as model substance to investigate the mechanism of xenobiotic metabolism in D. magna. The results of 24-h exposure experiments showed that D. magna could metabolize pyrene and biotransform it into water-soluble metabolites. On the other hand, the metabolism of pyrene was significantly inhibited by SKF-525A as the cytochrome P450 (CYP) inhibitor. These observations indicated that oxidation by CYP participated in the biotransformation of pyrene by D. magna. We also identified the pyrene metabolites formed by D. magna by HPLC with an electrospray ionization triple quadrupole mass spectrometry detector (LC/ESIMS/MS) and de-conjugation by sulfatase, beta-glucuronidase, and beta-glucosidase. One of the metabolites was ionized in ESI negative mode and formed a dominant mass of m/z 297 (MS) with the product ion of m/z 217 (MS2). Furthermore, this metabolite formed 1-hydroxypyrene on treatment with sulfatase. This metabolite was considered to be a sulfate conjugate of oxidized pyrene (1-hydroxypyrenesulfate). Furthermore, we quantified the deconjugated 1-hydroxypyrene formed by the above enzyme treatment. It showed that 52% of the total metabolized pyrene was biotransformed into 1-hydroxypyrene-sulfate, and more than 73% was biotransformed into oxidized pyrene conjugate. These results indicated that CYP and several conjugation enzymes participate in its biotransformation, and sulfation is important in D. magna for metabolism and elimination of xenobiotics.

This work describes the fundamental ability of a commercial ELISA to determine acetamiprid and the application of the ELISA to residue analysis in fruit and vegetable samples. The ELISA exhibited satisfactory sensitivity (I 50 0.6 ng/g; limit of detection 0.053 ng/g) and a high selectivity for acetamiprid versus other neonicotinoid analogs (thiacloprid amide). Methanol, which influenced the sensitivity of the ELISA the least, was selected as the extractant for the ELISA analysis. Simple dilution of sample extracts with water eliminated matrix interferences. Average recoveries from the acetamiprid-spiked agricultural samples were >95% using a simple extraction method. Analytical results obtained from the ELISA were comparable to those obtained from the reference HPLC method (r>0.99). The ELISA applied to the residue analysis of acetamiprid in agricultural products is a rapid, simple, and cost-effective method, and could be successfully applied to the detection of acetamiprid before the distribution of produce.

The performance of a commercially available enzyme-linked immunosorbent assay (ELISA) kit containing highly sensitive monoclonal antibodies against fenitrothion was assessed. The experimentally estimated dynamic range (0.087 to 2 ng/g) agreed with that established by the kit manufacturer (0.075 to 1 ng/g). The linearity of the standard curve produced for the kit-assembled standard solutions (slope = -0.3829) agreed with that of the curve produced for the self-made standard solutions (slope = -0.3619). The sensitivity (I 50 value) and the limit of detection for the kit were 0.23 and 0.033 ng/g, respectively. Selectivity of the ELISA indicates that the monoclonal antibody can readily distinguish the target pesticide from other structurally related analogs and some metabolites (oxon forms), with the exception of ethyl O-(p-nitrophenyl) phenyl phosphonothionate (EPN), parathion-methyl, and parathion. Methanol was the best organic solvent for the kit, with optimal sensitivity observed at a final concentration in the well of 10% (vol/vol) or less. Matrix interferences were minimized by direct dilution with water (60-fold) of the methanolic extracts from apple and peach samples. To extract fenitrothion from these two agricultural products as simply and rapidly as possible, three extraction methods were used. The extraction method that involved shaking by hand for 3 min was the best among the three methods. High recovery percentages (116.6% for apple and 110.8% for peach) were obtained. Validation of the ELISA method was carried out using the gas chromatography-mass spectrometry method, resulting in high recovery and close correlation of results (r > 0.95). These findings strongly suggest that the ELISA kit may be routinely employed for on-site fenitrothion screening of fruit samples.

Concentrations of esprocarb, thiobencarb, dimethametryn, pretilachlor and simetryn in water, suspended solids and sediment were studied during ordinary water discharge over the course of a year in Sugao marsh, Ibaraki Prefecture, Japan. Periodic changes in pesticide concentrations in water were similar to those in suspended solids. Pesticides were mostly present as dissolved form in water. Namely, the partition ratios of total suspended solids in whole water samples were 0.37–2.5% as the median, and the ratios of pesticide loading as adsorbed form on suspended solids against total pesticide loading were 0.42–4.6%. However, the concentrations of pesticides were higher in fine suspended solids (0.5 to 1.0 μm) than coarse suspended solids (more than 1.0 μm). Fine suspended solids loaded with pesticides were estimated to be 18 to 59% of all suspended solids, indicating their important role as a carrier of pesticides. Moreover, vertical distributions of pesticides in sediment down to a depth of 15 cm were not clearly shown, probably due to the disturbance of sediment at the surface by actual water flow. © Pesticide Science Society of Japan

Article, Enviromental Pollution. 138(3): 529-537 (2005)

The development of an enzyme-linked immunosorbent assay (ELISA) based on polyclonal antibodies for a class of endocrine disrupting compounds, 4-nonylphenol, is described. The parent molecule was derivatized at the ortho position of the free phenolic hydroxyl group to obtain the hapten, NP1, and it was conjugated with keyhole limpet hemocyanin, which was used as an immunogen. Four antisera were generated and screened against three coating antigens. The most sensitive ELISA from the screening tests (antiserum NP03As, 1/1000, and coating antigen NP1-BSA, 1 microg/mL) was further optimized and characterized. The influence of various physicochemical factors (organic solvent, pH, ion strength) was investigated. Methanol as the additive organic solvent was found to be the best organic solvent for the ELISA, with optimal sensitivity observed at a concentration of 5%. The ELISA parameters were changed at more acidic or basic pH values, whereas higher ionic strengths strongly suppressed the I(50) value and the maximum absorbance. The most sensitive ELISA for 4-nonylphenol exhibited an I(50) value of 38.6 +/- 5.5 microg/L, with a dynamic range from 12 to 350 microg/L, and the lower limit of detection was 7.7 +/- 1.3 microg/L. The optimized ELISA displayed no significant cross-reaction against the parent compounds, nonylphenol ethoxylates, degradation products, carboxylates, and bisphenol A, except in 4-octylphenol.

A recently developed monoclonal antibody-based ELISA kit for neonicotinoid insecticide imidacloprid was evaluated for its reproducibility, accuracy, and comparability to results from a conventional high-performance liquid chromatography (HPLC) technique for the determination of residual imidacloprid in agricultural products (cucumber, eggplant, lettuce, and green pepper). A method in which imidacloprid is extracted from each agricultural sample as simple, rapid, and efficient as possible was examined for application to screening field analysis, finally. When each extract was diluted 10-fold and over with water prior to ELISA determination, no significant matrix interference was observed besides spinach. The recovery values from fortified samples and reproducibility were very good (average recovery values ≥85.3%; CV values ≤24.0%). The correlation between methods was excellent ( r ≥0.91). The ELISA kit is a suitable screening method for quantitative and reliable determination of imidacloprid in vegetables even without sample clean-up, which saves time and money and considerably increases sample throughput.

A l -leucine-seed-modified quartz crystal microbalance (QCM) oscillator showed selectively chiral discrimination in situ for leucine enantiomers. The obtained results exhibited strong QCM response to l -leucine, but the response to d -leucine was negligible. This selectivity was attributed to preferential crystallization of l -leucine at the solid-solution interface.

A quartz crystal microbalance (QCM) sensor for selenite ions in aqueous solution was constructed based on crystal formation of cadmium selenite, immobilized with a self-assembly monolayer (SAM) of phosphorylated 11-mercapto-1-undecanol (MUD) on a QCM gold electrode surface. The mass change caused by the selective adsorption of selenite ions on the cadmium selenite crystals at the solid/solution interface was detected by the QCM. The response (−ΔF) of the modified QCM oscillator increased with increasing selenite ion concentrations in sample solutions, ranging from 9.7×10−5 to 9.0×10−4 M at pH 7.4. The synthetic process of anchoring cadmium selenite crystals on the phosphorylated MUD organic film was also followed by using X-ray photoelectron spectroscopy (XPS) and atomic force microscopy (AFM). The atomic concentrations measured by XPS confirmed the crystal growth of cadmium selenite on the phosphorylated MUD SAM at the QCM gold electrode surface. From the AFM images, changes in surface topographic features were followed: the MUD SAM and phosphorylated MUD on the QCM gold electrode had similar surface roughness; however the difference for the cadmium selenite film on the phosphorylated MUD SAM was clearly seen. The observed QCM frequency change of the modified QCM oscillator per unit time was found to be proportional to the square of the supersaturation of cadmium selenite, indicating the crystal growth of cadmium selenite at the solid/solution interface. The modified QCM oscillator exhibited selectively strong QCM response to SeO32− ion. In contrast, the responses to tested interfering anions were almost negligible. The order of anion selectivities of the present modified QCM sensor was SeO32−≫CO32−>SeO2−4, SO42−, Br−, I−, NO3−. These selectivities were basically attributable to the differences in solubility products and solubilities for the salts of each anion with cadmium (II) ion.

A self-assembly monolayer (SAM) of phosphorylated 11-mercapto-1-undecanol (MUD) on a quartz-crystal microbalance (QCM) gold electrode was served as an interfacial layer for subsequent adhesion of barium sulfate crystals. The subsequent process of anchoring barium sulfate crystals on the phosphorylated MUD organic film was followed using X-ray photoelectron spectroscopy (XPS) and atomic force microscopy (AFM). The atomic concentration measured by XPS supported the growth of barium sulfate crystal on the phosphorylated MUD SAM on the QCM gold electrode surface. The AFM image of the barium sulfate crystals grown on the phosphorylated MUD SAM showed a smaller surface roughness than those of the MUD and of the phosphorylated MUD SAM. The observed QCM frequency change per unit time of the modified QCM oscillator was found to be proportional to the square of the relative supersaturation of barium sulfate. This also indicated the crystal growth of barium sulfate at the solid/solution interface. The response (−Δ F ) of the modified QCM oscillator increased with increasing sulfate ion concentrations in adjacent sample solutions ranging 6.8×10 −5 –4.5×10 −3 M at pH 7.2. These responses were due to the selective adsorption of sulfate ions on the barium sulfate crystals at the solid/solution interface. The relative standard deviation was found to be less than 3%. The order of relative anion selectivities for the QCM frequency change of the modified QCM oscillator was: sulfate ion>carbonate ion≫NO − 3 , I − , Br − , SCN − . These relative selectivities were attributed to the differences in solubility product and solubilities for the salts of each anion with barium(II) ion.

The potentiometric responses of copper and nickel hexacyanoferrate membrane electrodes were examined for alkali, alkaline earth, heavy metal and ammonium ions. When K2Cu3[Fe(II)(CN)6]2 membranes were electrodeposited on a Cu plate in aqueous K4[Fe(II)(CN)6] solution at applied potentials from +0.20 to +0.40 V vs SCE, the membranes exhibited a near-Nernstian response (55 mV/decade) to K+ ions above 1×10−4 M. When this K2Cu3[Fe(II)(CN)6]2 membrane was immersed and conditioned in 0.1 M (CH3)4N·NO3 (tetramethylammonium nitrate, TMA·NO3) solution prior to potentiometric measurements, the preferential dissolutions of K+ ions into the adjacent solution was observed accompanying negative membrane potential shifts (more than 200 mV). The potentiometric response of copper hexacyanoferrate membranes electrodeposited above 0.40 V vs SCE became however weaker with decreasing K+ content in CuHCF membranes and both the amount of preferential K+ dissolution and the extent of the membrane potential shift became smaller during the membrane conditioning. From these results, it was concluded that negatively charged K+ vacancies at the membrane surface contacting the adjacent electrolyte solution (0.1 M (CH3)4N·NO3) were formed by the preferential K+ dissolution, into which the analyte K+ ions was back-titrated during the potentiometric response process. The potentiometric selectivities of the K2Cu3[Fe(II)(CN)6]2 and KNi[Fe(III)(CN)6] membrane electrodes for alkali metal ions were Cs+>Rb+>K+>Na+>Li+ and no significant responses were observed for any alkaline earth metal ions in the concentration range below 1.0×10−2 M. These selectivities may reflect their dehydration energies needed for their accommodation from the solution side into the negatively charged vacancies formed by the preferential K+ dissolution at the membrane surface. The potentiometric response to a series of ammonium ions was in the following order; NH4+>(CH3)NH3+>(CH3)2NH2+≫(CH3)3NH+, (CH3)4N+, CH3CH2NH3+, (CH3CH2)2NH2+, (CH3CH2)4N+, (CH3)CNH3+, CH3C3H6NH3+, CH3C3H6NH2+CH3, (CH3)2CHCH2NH2+CH3, reflecting the size of the analyte ions. On the contrary, the response to divalent heavy metal cations (selectivity; Cu2+>Pb2+>Zn2+>Mn2+>Ni2+>Cd2+) however seemed to be due to specific adsorption onto the solid surface rather than the uptake of relevant ions into the vacancies formed by the preferential K+ dissolution. The latter mechanism appeared to be similar to the one previously reported for conventional precipitated-based ISEs, such as CuS, CdS and AgX (X=Cl, Br and I).

To reduce hazardous organic solvent consumption during sample preparation procedures as much as possible, an extraction method of smallest feasible sample volume (5g) using aqueous acetonitrile (MeCN) was developed to extract pesticide residues from agricultural samples prior to HPLC-DAD determination. Extraction with MeCN/water (1:1, v/v), and adjustment of the MeCN concentration by diluting with water after extraction recovered successfully most pesticides showing various physicochemical properties. The matrix effects of tested samples on the proposed method developed herein were generally negligibly-small. The average recoveries were in the range 70-120% for all pesticides with the coefficient of variation values below 20%. The reduction rate of organic solvents used for the proposed sample preparation method was up to approximately 60% compared with the Japanese authorised official method for pesticide residue analyses. These results demonstrate the feasibility of the proposed method for pesticides with diverse properties.

An original extraction method using water as an extractant has been established for environmentally friendly sample preparation procedures for hydrophilic pesticides (acetamiprid, clothianidin, dinotefuran, flonicamid, imidacloprid, methomyl, pymetrozine, thiacloprid, and thiamethoxam) in agricultural samples with conventional HPLC. Water-based extraction and cleanup with two solid-phase extraction cartridges can recover target hydrophilic pesticides quantitatively. The matrix effects of tested samples on the proposed method developed herein were negligibly small. Under the optimized conditions, the recoveries of almost all tested pesticides were 70-120% with satisfactory precision (%CV20%). The analytical data are in good accordance with Japanese or European Union guidelines for pesticide residue analysis. The reduction rate of hazardous organic solvents used for the proposed method and by reducing the sample size for extraction was about 70% compared with the Japanese authorized reference method used in this work. The results demonstrate the feasibility of the proposed sample preparation procedures for hydrophilic pesticides.

Zucchini cultivars Cucurbita pepo subsp. ovifera cv. Patty Green and subsp. pepo cv. Gold Rush were cultivated hydroponically in a nutrient solution supplemented with a mixture of dioxins and dioxin-like compounds. Patty Green and Gold Rush showed low and high accumulation of these compounds in the aerial parts respectively. In both cultivars, the accumulation of each congener negatively depended on its hydrophobicity. This suggests that desorption and solubilization were partly responsible for congener specificity of accumulation, since this was not found in soil experiments. In contrast, no clear difference in accumulation in the roots was observed between the cultivars, whereas the translocation factors, which are indicators of efficient translocation from the roots to the aerial parts, differed among the congeners hydrophobicity-dependently. There were positive correlations between accumulation in the roots and the hydrophobicity of the polychlorinated biphenyl congeners in both cultivars. These results indicate that translocation was also partly responsible for the congener specificity and accumulation concentrations.

Diphenylarsinic acid, phenylarsonic acid, methylphenylarsinic acid (MPAA), dimethylphenylarsine oxide (DMPAO), and methyldiphenylarsine oxide (MDPAO) in soil and rice were extracted, separated by reversed-phase chromatography, and quantified by ICPMS with a membrane desolvating system. For the extraction of arsenicals from rice grain and straw, 68% HNO3 provided better extraction efficiency than water, 50% methanol, or 2.0 mol L(-1) trifluoroacetic acid. For the extraction from soil, 68% HNO3 provided better extraction efficiency than H2O, 1 mol L(-1) H3PO4, or 1 mol L(-1) NaOH. The contaminated soil contained all five aromatic arsenicals along with inorganic arsenicals as main species (5.86 +/- 0.19 microg of As kg(-1): 60.8 +/- 2.0% of total extracted As). After pot experiments, rice straw contained mainly DMPAO (7.71 +/- 0.48 microg of As kg(-1): 60.5 +/- 3.7%), MDPAO (0.91 +/- 0.07 microg of As kg(-1): 7.2 +/- 0.5%), and inorganic As (2.85 +/- 0.20 microg of As kg(-1): 22.3 +/- 1.6%). On the other hand, rice grain contained mainly MPAA (1.17 +/- 0.04 microg of As kg(-1): 86.7 +/- 2.7%). The root uptake of each species from the soil and transport from straw to grains were significantly related to the calculated log K(ow) values.

The cholinesterase (ChE)-inhibiting activity of water and the concentrations of representative inhibitors were monitored in the Tone canal, Japan, during April to December 2006. The ChE-inhibiting activity, measured by using horse serum as enzyme source, increased from late April to early June, and from September to October. Although the trends in the ChE-inhibiting activity of the samples were consistent with concentration changes of organophosphorus pesticides, ChE-inhibiting activity was not observed in samples replicated on the basis of the chemical concentrations detected. The water samples were treated with chlorine to enhance the ChE-inhibiting activity by conversion of thiophosphate pesticides to phosphate pesticides. The ChE-inhibiting activity increased in almost all the chlorine-treated samples, although organophosphorus pesticides were either not detected or detected in traces in the samples by gas chromatographic–mass spectrometric analysis. These results suggested that assay of ChE-inhibiting activity is important for evaluating the ecotoxicity of environmental water, because toxicological investigations based solely on inhibitor concentrations may underestimate the contamination. Furthermore, the combined method of oxidation by chlorination and the ChE assay is very effective for screening and monitoring of organophosphorus pesticides in environmental water.

We found that aquatic crustaceans, decapoda; atyidae (Caridina multidentata, Neocaridina denticulate, and Paratya compressa), metabolize pyrene to a new conjugation product. The results of deconjugation treatments indicated that glucose and sulfate combined with 1-hydroxypyrene. Further analysis by LC/ESI-MS/MS showed that the molecular weight of the product was 460 (m/z 459; deprotonated ion), and that it has a glucose-sulfate moiety (m/z 241; fragment ion). These results indicated that the new metabolite was the glucose-sulfate conjugate of 1-hydroxypyrene. The glucose-sulfate conjugate is a phase II product that has not been reported previously from any organism. Several studies have demonstrated that sulfation is an important pathway for metabolism of xenobiotics in aquatic invertebrates. Thus, glucose-sulfate conjugates may add an important signal for excretion or sequestration of xenobiotics for aquatic invertebrates.

The zucchini (Cucurbita pepo) cultivars 'Patty Green', 'Black Beauty', and 'Gold Rush' were cultivated on weathered dioxin-contaminated soil in pots, and concentrations of the 29 dioxin-like compounds that were assigned WHO-TEFs, three non-toxic polychlorinated dibenzo-p-dioxins/dibenzofurans (PCDD/Fs), and two non-dioxin-like polychlorinated biphenyls (PCBs) were analyzed. Toxic equivalent (TEQ) values accumulated in 'Black Beauty' and 'Gold Rush' were about 180 times higher than those in 'Patty Green'. The bioconcentration factor (BCF) based on total mass concentration of the twelve dioxin-like PCBs was higher than those of the seven PCDDs and ten PCDFs in all the cultivars. The BCFs for PCDD and PCDF congeners were negatively correlated with octanol-water partition coefficients in all the plants. No correlations were observed in PCB congeners in the high accumulators, although in 'Patty Green' the BCFs for PCB congeners were significantly correlated with octanol-water partition coefficients. Our findings suggest that the high accumulators had unknown, unique mechanisms for uptake of PCBs, whereas PCDDs and PCDFs were absorbed based on their physicochemical properties.

Thirteen isomers of branched para-nonylphenols (para-NP) in three technical mixtures were isomer-specifically determined using their synthesized standards by SIM of structurally specific ions, m/z 135, 149 or 163 with GC–MS. Of the 13 isomers, four isomers, 4-(2,4-dimethylheptan-4-yl)phenol, 4-(4-methyloctan-4-yl)phenol, 4-(3-ethyl-2-methylhexan-2-yl)phenol (3E22NP) and 4-(2,3-dimethylheptan-2-yl)phenol synthesized for their determinations were first used as standard substances. The 13 isomers in the technical mixtures individually occurred at mass percent portion of more than 2%. The total mass percent portions in the mixtures from Tokyo Chemical Industry (TCI), Aldrich, and Fluka covered with 89 ± 2%, 75 ± 4% and 77 ± 2%, respectively. The abundance of 4-(3,6-dimethylheptan-3-yl)phenol in the three mixtures was the largest with 11.1 ± 2% to 9.9 ± 0.3%, while that of 4-(2-methyloctan-2-yl)phenol was the smallest with 2.9 ± 0.3% to 3.0 ± 0.2%. Additionally, structures of four new isomers of more than 1% portion present in a technical mixture were elucidated as two pairs of diastereomeric isomers: two types of 4-(3,4-dimethylheptan-4-yl)phenol (344NP) and those of 4-(3,4-dimethylheptan-3-yl)phenol (343NP). By estrogenic assay of 13 isomers with yeast estrogen screen system, the activity of 3E22NP was the highest, while that of 4-(3-methyloctan-3-yl)phenol was the least. Their relative activities to that of 3E22NP were individually calculated. Estrogenic equivalent concentrations of the three technical mixtures were predictively evaluated. The ratio of the EEC to the conventional concentration, total mass percent portions of the 13 isomers in technical mixtures were 0.208 for TCI, 0.206 for Aldrich and 0.205 for Fluka. The predicted estrogenic activity of measured concentration of para-NP in technical mixtures was approximately 5-fold greater than the measured estrogen agonist activity.

Persistent organic pollutants (POPs) of organochlorine pesticides such as dichlorodiphenyl trichloroethane (DDTs) and its metabolites, hexachlorobenzene (HCB), heptachlor, chlordane compounds (CHLs), aldrin, dieldrin, endrin, mirex and isomers of hexachlorocyclohexane (HCHs), were analyzed in sediment cores collected from Ariake Bay, Japan. Although Chikugo River has the largest flow in Ariake Bay, the total concentration of POPs in surface sediment was found in St.3, mouth of the Yabe River, where that level was two times higher than St.1, mouth of the Chikugo River. It could be assumed that the potential source of POPs contamination is relatively close to this study area. Relatively high residue levels of HCB, HCHs and DDTs in sediment core were found during the period from 1967 to 1970. In this layer, the HCB concentration investigated in sediment cores of Ariake Bay was maximum, 2.6 ng g(-1) dry weight and higher than that of Tokyo Bay and subsequently detected in deeper sediment cores. The ratio of (DDD+DDE)/SigmaDDTs was high in top sediments (0-2 cm). Although there was a concentration of DDTs in the top sediment, it was noticed that DDT emission did not cause direct input of DDT recently. The HCHs determined in upper sediment cores might be originated from long-range accumulation in environment after dispersing of technical-grade HCH. Moreover, the CHLs, dieldrin and heptachlor concentrations were detected and recently increased in sediment core. POPs in sediment cores of Ariake Bay based on the possibility of PCP contamination during the early 1960s were reflected.

This work describes the analytical performance of the enzyme-linked immunosorbent assay (ELISA) for the fungicide chlorothalonil to effectively exploit as a simple and rapid detection system for pesticide residue on the scenes of the agricultural production and distribution. This ELISA represents the satisfactory analytical characteristics (I50 value, 0.34 ng/g; limit of detection, 0.052 ng/g) to detect chlorothalonil at the regulatory values or thereabout in a sample. Noticeable cross-reactivities were shown with two fungicides, fthalide (58.8%) and pentachloronitrobenzene (quintozene) (20.0%), and some non-agrochemicals such as tetrachloroterephthalonitrile (96.8%) and tetrachlorophthalonitrile (68.3%). The influence of three organic solvents (methanol, acetone, and acetonitrile) used as extractants for chlorothalonil residue was evaluated, with the result that methanol was the most suitable solvent for the ELISA, and the final concentration in the well could be up to 5% (v/v) without any negative influence on the ELISA. It has been possible to directly analyze chlorothalonil residue only by giving dilution of each sample extract with water prior to the ELISA analysis. The average recovery values from the spiked samples by the ELISA were between 101.7 and 113.6% with the average coefficients of variation between 2.6 and 5.9%. Although the results obtained from the ELISA correlated well with those from the reference GC/MS methods for all agricultural samples (r > 0.98), the linear function inclined to the ELISA results because of loss during complex sample preparations for GC/MS analysis. Nevertheless, the results demonstrated that the proposed ELISA is a reliable, cost-effective, and rapid quantitative method for chlorothalonil residue.

A commercially available enzyme-linked immunosorbent assay (ELISA) kit with a high affinity monoclonal antibody was applied to residual analysis of insecticide chlorfenapyr in agricultural samples, and drawn a parallel between the ELISA and gas chromatography (GC) with mass spectrometry (MS). For standards prepared in water containing 5% (v/v) methanol, the sensitivity (I50 value), the dynamic range, and the limit of detection of the ELISA kit were 2.3, 1 - 10, and 0.1 ng/g, respectively. The used monoclonal antibody in the ELISA kit had a high selectivity. The ELISA kit was applied to the determination of chlorfenapyr in two kinds of fruits (apple and peach). The examination of the influence of these matrices on the reliability of the assay performance indicated that the ELISA could determine it in these samples near the regulation values in Japan simply by diluting the methanolic extract or by concentrating it, without any clean-up procedures. Recovery and precision of the proposed ELISA method were assessed by fortifying fruit samples with chlorfenapyr ranging from 0.05 to 1.5 microg/g. Mean recoveries were 94.2 and 90.3% for apple and peach, and coefficients of variation were below 16% in most cases. The results obtained from the proposed ELISA method correlate well the reference GC/MS method for both fruit samples (r > 0.98). These considerations make the ELISA kit very useful analytical tool for monitoring and regulatory programs, without the need of complex and expensive instrumentation.

The performance of a commercially available microtiter plate ELISA kit for the determination of the neonicotinoid insecticide imidacloprid was evaluated for sensitivity, selectivity, influence of organic solvent used for extraction procedure, matrix interference originated from agricultural sample, accuracy, and method comparison with conventional HPLC analysis. The limit of detection for the kit (0.1 or 0.5 ng/mL) was determined. The working range (1-39 ng/mL) experimentally calculated on the basis of a criterion, which is determined as the range from I(20) to I(80), was comparable to that established by the manufacturer (1-50 ng/mL). The linearity of the standard curve based on the kit-assembled standard solutions agreed with the one based on the self-made standard solutions. Specificity studies indicate that the imidacloprid monoclonal antibody can readily distinguish the target compound from other structurally related neonicotinoid analogues and some metabolites, with the exception of clothianidin, the cross-reactivity of which was approximately 12%. To extract imidacloprid from an agricultural sample (apple) as simply and rapidly as possible, some extraction methods were examined. Consequently, the extraction method with hand-shaking for 5 min was the best among the examined methods. For the analysis of imidacloprid in apple samples, it was extracted directly with methanol and the extracts were diluted 10-fold (100-fold in the well) with water prior to ELISA analysis. No significant matrix interference was observed with the dilution factor. Recoveries of imidacloprid from fortified apple samples ranged from 87.7 to 112.0%. The results obtained with the ELISA kit correlated well with those by the reference method (conventional HPLC analysis) for apple samples (r0.998). These findings strongly indicate that the ELISA kit may be employed routinely for an on-site imidacloprid residue analysis of apple samples.

Dioxin concentrations and homologue profiles were examined in plucked new shoots, crude tea and its hot water extracts, soils and atmosphere in tea orchards. The rate of dry provisions in crude tea has increased 4 times as that of the plucked new shoots. However, dioxin concentrations except O8CDD in crude tea have increased only 2 to 3 times as those of the plucked new shoots. O8CDD concentration increased remarkably during processing. Little dioxins were detected in hot water extracts of crude tea leaves.

Direct determination of cadmium in rice flour by laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) is described. This superior quantitative method needs no conventional laborious sample pretreatment such as decomposition and extraction. To construct the calibration line and estimate the detection limit, three certified reference materials for rice flour, NIES nos. 10a, 10b and 10c, were used and contaminated with three different concentrations of cadmium (0.023, 0.32 and 1.82 mg kg−1, respectively). These three kinds of rice flour were finely ground and pressed into disks for analysis. The calibration line showed good linearity in the range 0.023–1.82 mg kg−1. The determination did not require internal standardization and no significant interference was observed.