Japan Flavour and Fragrance Materials Association's (JFFMA) safety assessment of food-flavouring substances uniquely used in Japan that belong to the class of aliphatic primary alcohols, aldehydes, carboxylic acids, acetals and esters containing additional oxygenated functional groups.

We performed a safety evaluation using the procedure devised by the Joint FAO/WHO Expert Committee on Food Additives (JECFA) of the following four flavouring substances that belong to the class of 'aliphatic primary alcohols, aldehydes, carboxylic acids, acetals, and esters containing additional oxygenated functional groups' and are uniquely used in Japan: butyl butyrylacetate, ethyl 2-hydroxy-4-methylpentanoate, 3-hydroxyhexanoic acid and methyl hydroxyacetate. Although no genotoxicity study data were found in the published literature, none of the four substances had chemical structural alerts predicting genotoxicity. All four substances were categorised as class I by using Cramer's classification.

The JFFMA assessment of flavoring substances structurally related to menthol and uniquely used in Japan.

Using the procedure devised by the Joint FAO/WHO Expert Committee on Food Additives (JECFA), we performed safety evaluations on four flavoring substances structurally related to menthol (L-menthyl 2-methylbutyrate, DL-menthyl octanoate, DL-menthyl palmitate, and DL-menthyl stearate) uniquely used in Japan. While no genotoxicity study data were available in the literature, all four substances had no chemical structural alerts predictive of genotoxicity. Moreover, they all four are esters consisting of menthol and simple carboxylic acids that were assumed to be immediately hydrolyzed after ingestion and metabolized into innocuous substances for excretion.

[Residual pesticide concentrations after processing various types of tea and tea infusions].

The effects of processing to produce various types of tea or infusion on the levels of pesticide residues in tea were investigated for three insecticides (chlorfenapyr, pyrimiphos-methyl, and clothianidin). Tea plants were sprayed with one of the three pesticides and cultivated under cover. The levels of pesticide residues in tea decreased after processing according to the time and temperature of heating, as well as fermentation.

Analysis of trans-fat levels in total diet and one-serving samples using the verified GC-method and estimation of the intake in Japan.

In Japan, discussions on the regulation and labeling of trans-fat (TF) have under way for several years in the Food Safety Commission and the Consumer Affairs Agency. However, administrative measures for TF have not yet been taken, partly because of the insufficiency of scientific data in Japan. To provide data about the TF intake by Japanese, we determined the levels of TF contained in total diet samples and in food samples that were served as individual meals (one-serving samples).

Determination method for total arsenic and partial-digestion method with nitric acid for inorganic arsenic speciation in several varieties of rice.

The determination method of total arsenic (As) and the speciation method of inorganic As for non-glutinous rice reported in our preceding paper were applied to several varieties of rice under optimized experimental conditions. In the determination of total As with ICP-MS, acetic acid was added to increase the sensitivity and an internal reference method with germanium was adopted to increase the precision. The extraction temperature in the partial-digestion method with nitric acid to speciate inorganic As was raised to 100 degrees C, because extraction efficiency over 90% was obtained from glutinous rice and colored rice at this temperature.

[Surveillance of perchlorate level in leafy vegetables and bottled water].

Perchlorate (ClO(4)(-)) is both a naturally occurring and artificial compound, and it inhibits iodide uptake into the thyroid gland and disturbs thyroid function. It has been detected in many foods in the United States. In order to investigate perchlorate contamination in foods in Japan, perchlorate level in 82 leafy vegetable samples and 20 bottled mineral water samples was measured using a procedure based on the FDA's procedure, employing IC-MS/MS with (18)O(4)-labeled perchlorate as an internal standard.

[Analysis of demeton-S-methyl, oxydemeton-methyl and demeton-S-methylsulfone in agricultural products by LC-MS].

We studied the simultaneous determination of demeton-S-methyl, oxydemeton-methyl, and their oxide demeton-S-methylsulfone, in agricultural products by liquid chromatography coupled with mass spectrometry (LC-MS). The sample homogenized with antioxidants L-ascorbic acid and butylhydroxytoluene was extracted with acetone. An aliquot of the crude extract was reextracted with ethyl acetate by using an EXtrelut column.

[Analytical method of dodine in agricultural products by LC/MS].

A method for the determination of dodine in agricultural products was developed by using liquid chromatography-electrospray ionization mass spectrometry (LC/ESI-MS). Dodine was extracted with acetonitrile and then acetonitrile-water (7 : 3) from a sample, and re-extracted with ethyl acetate. The extract was cleaned up on a PSA cartridge column (500 mg), and dodine was analyzed by LC/MS.

Evaluation of modified PCR quantitation of genetically modified maize and soybean using reference molecules: interlaboratory study.

Real-time polymerase chain reaction (PCR)-based quantitative methods were previously developed and validated for genetically modified (GM) maize or soy. In this study, the quantification step of the validated methods was modified, and an interlaboratory study was conducted. The modification included the introduction of the PCR system SSIIb 3 instead of SSIIb 1 for the detection of the taxon-specific sequence of maize, as well as the adoption of colE1 as a carrier included in a reference plasmid solution as a replacement for salmon testis.

[Estimation of the daily intake of nitrate based on analysis of total diet samples].

The concentrations of nitrates in total diet samples prepared at 11 places were determined according to the market basket method, and the daily intake of nitrates was estimated based on the analysis results. The mean estimated daily intake of nitrates was 200 mg/man/day (4.0 mg/kg/day) and exceeded the ADI (3.

[Factors affecting the variance of copy numbers in real-time PCR as estimated from calibration curves].

The distribution interval of calibration curves in real-time PCR was successfully predicted from the variance of the Ct values, and the predicted distribution interval was consistent with the interval of the actual calibration curves. Although the width of the 95% confidence interval at the lower end was about 1%, the copy number estimated from individual calibration curves was distributed over a wider range, and the 95% confidence interval of the estimates of 20 copies was +/-40%. The variances of the Ct values at 20 copies were significantly larger than those at higher copy numbers, and this large variation increased the distribution interval of the calibration curves.

[Development and evaluation of determination methods for boric acid in agar using ICP-AES and ICP-MS].

A new test method for boric acid in agar was developed. After digestion with nitric acid, the concentration of boron was measured by ICP-AES or ICP-MS with internal standards. Collaborative studies involving 5 laboratories were conducted to evaluate the new method by using a finely powdered agar sample and a standard sample (NIST SRM SRM1570a).

Orally administered apple procyanidins protect against experimental inflammatory bowel disease in mice.

Apple procyanidins (ACT) is a natural biologically active compound extracted from apple. Our recent studies have shown that ACT ameliorates the symptoms of atopic dermatitis and inhibits food-allergen-induced oral sensitization. The aim of this study was to investigate the potential protective effect and mechanism of action of ACT in a murine model of inflammatory bowel disease.

[Effect of non-genetically modified (non-GM) soy varieties on the measured value of GM soy by a quantitative PCR method].

It is very important to examine the effect of non-genetically modified (non-GM) soy varieties, which constitute the matrix of the testing sample used to quantify GM soy (RRS), on the measured value of RRS by quantitative PCR methods. Therefore, we quantified the amount of RRS in powder-mixed samples containing 1 or 5% RRS prepared by using 10 different varieties of non-GM soy as the matrix. The results revealed that the measured values were not in agreement with the powder-mixing levels and that the extent of the difference depended on the variety of non-GM soy used as the matrix.

Simultaneous determination of sedecamycin and terdecamycin in livestock products by LC/MS.

A simple and rapid LC/MS method for simultaneous determination of sedecamyin (SCM) and terdecamycin (TDM) in livestock products has been developed. SCM and TDM were extracted with acetonitrile. The extract was washed with n-hexane and then evaporated to dryness.

Evaluation of a nitric acid-based partial-digestion method for selective determination of inorganic arsenic in rice.

Arsenic (As) uptake in human occurs via the food chain mainly. The Joint FAO/WHO Expert Committee on Food Additives has established the provisional tolerable weekly intake level for As as an inorganic As (iAs) value, because iAs in food is much more toxic than organic As. In this study, we studied an acid based partial-digestion method for the complete extraction of arsenicals from rice.

Nitric acid-based partial-digestion method for selective determination of inorganic arsenic in hijiki and application to soaked hijiki.

Because there is a great difference between the toxicity of inorganic arsenic (As) and organic As in food, the JECFA has set a PTWI value for inorganic As (iAs) rather than for total As. The difference in As toxicity makes it necessary to extract iAs completely from food samples for toxicological analysis, but complete extraction of As from most foods including seaweed has not been achieved to date. We developed a partial-digestion method that uses nitric acid as a solvent in order to extract almost all arsenicals from the solid matrix of hijiki (Hizikia fusiforme, a brown alga) samples.

DNA extraction method using a silica-base resin type kit for the detection of genetically modified papaya.

Genetically modified (GM) papaya has not yet been approved for importation into, or cultivation in the European Union (EU) and Japan. A DNA extraction method using the Qiagen DNeasy Plant Mini Kit (PM method) and a method using a buffer containing cetyltrimethyl ammonium bromide (CTAB method) have been adopted as the official Japanese methods for detecting GM foods. However, the amounts of DNA extracted from papaya by these methods are very low.

Biosensor immunoassay for the screening of dioxin-like polychlorinated biphenyls in retail fish.

Dioxin-like polychlorinated biphenyls (DL-PCBs) often make up the majority of the toxic equivalent (TEQ) contribution of dioxins found in fish samples. For the purpose of making risk assessments, it is therefore important to develop screening methods for determining TEQ concentrations of DL-PCBs in retail fish. We have developed a rapid biosensor immunoassay (BIA) for DL-PCBs that uses a surface plasmon resonance sensor (Biacore 3000).

PCB 118 and Aryl Hydrocarbon Receptor Immunoassays for Screening Dioxins in Retail Fish.

The efficacy of a combination of two enzyme-linked immunosorbent assay (ELISA) kits was examined for screening the toxic equivalent (TEQ) concentrations of dioxins in retail fish. The coplanar PCB-EIA system, which is a competitive immunoassay specific for polychlorinated biphenyl (PCB) 118, was tested as a screening method for mono- ortho PCBs. The Ah immunoassay (Ah-I), which is an ELISA-based aryl hydrocarbon receptor binding assay, was analyzed for its screening ability for non- ortho PCBs, polychlorinated dibenzo- p-dioxins (PCDDs), and dibenzofurans (PCDFs).

Interlaboratory evaluation of two enzyme-linked immunosorbent assay kits for the determination of crustacean protein in processed foods.

The labeling of foods containing material derived from crustaceans such as shrimp and crab is to become mandatory in Japan because of increases in the number of allergy patients. To ensure proper labeling, 2 novel sandwich enzyme-linked immunosorbent assay (ELISA) kits for the determination of crustacean protein in processed foods, the N kit (Nissui Pharmaceutical Co., Ltd, Ibaraki, Japan) and the M kit (Maruha Nichiro Holdings, Inc.

[Method for DNA extraction and purification from corn-processed food using an ion-exchange resin type kit].

The yield of genomic DNA extracted from corn-processed foods, such as corn flake and one of the corn snack what is called "Jumbo corn", using an ion-exchange resin type kit (Gtip) has been reported to be very low, and it is thought to be difficult to detect the intrinsic corn gene "Zein" in the foods. Therefore, we developed a new method using Gtip, which we called the "KNG-Gtip method," by modification of the Ministry of Health, Labour and Welfare (MHLW) method using Gtip (MHLW-Gtip method). We compared the KNG-Gtip method, MHLW-Gtip method, the Gtip method for detection of allergen (ALG-Gtip method), and the Gtip method according to the Ministry of Agriculture, Forestry and Fisheries (MAFF) (JAS-Gtip method) in terms of the yield and quality of genomic DNA and the detection probabilities of the PCR-amplified Zein gene.