An official analytical method for chlorophyll degradation compounds, including pheophorbide, in chlorella products, is described in notification Kanshoku No. 99 (May 8, 1981). However, this method has several operational issues, such as the formation of emulsion during liquid-liquid partitioning. Additionally, impurities present in the reagents (sodium sulfate decahydrate or anhydrous sodium sulfate) used to prepare saturated sodium sulfate solution can degrade pheophorbide and other related compounds, resulting in a significant decrease in analytical values. In this study, we thoroughly examined each step of the official method to enhance the operability and develop an alternative method that eliminates the need for saturated sodium sulfate solution. The developed method was evaluated for pheophorbide a and pyropheophorbide a at 100 mg%. Satisfactory analytical performance was achieved with trueness of 100% for pheophorbide a and 90% for pyropheophorbide a, and relative standard deviations of intra- and inter-day precision below 5% for both compounds. The proposed method is considered suitable for regulatory analysis of chlorophyll degradation compounds and would be useful for quality control of chlorella products.

The maximum growth rate (μ_max) of Bacillus cereus was estimated using a non-destructive isothermal calorimetric method, and a growth prediction model was constructed based on the measurement results. SCD medium and mashed potato were inoculated with serial-diluted inoculum of B. cereus. Heat generation curves were determined using an isothermal calorimeter at 35, 25, and 15℃. The μ_max was determined from the relationship between the increase in B. cereus cell number and incubation time, which was detected through the heat generation of the B. cereus biological process. Moreover, the growth prediction model was constructed using Ratkowsky’s square-root model. The results of the growth prediction model based on the data of the calorimetric and conventional culture methods for SCD were expressed as √μ^Cal_max=0.0354 (T-4.9)[R²=0.99] and √μ^CCM_max=0.0335 (T-5.0)[R²=0.99]; a similar equation was provided by both methods. Conversely, the results of the growth prediction model based on the calorimetric method data for mashed potato were given as √μ^Cal_max=0.0390 (T-8.5)[R²=0.99]; the maximum growth rates at 30 and 20℃ were predicted as 0.70 and 0.20 (1/hr), respectively. The maximum growth rates obtained using the conventional culture method were 0.63 and 0.29 (1/hr), respectively, similar to the calorimetric method results. The predictive microbiological analysis using the calorimetric method enabled the rapid provision of a growth prediction equation, and the number of samples could be substantially reduced compared with that for the conventional culture method.

Irradiation is widely used worldwide to sterilize and kill insects in food, and prevent the germination of agricultural products. However, in Japan, food irradiation is prohibited except to prevent potato sprouting. Herein, 5,6-dihydrothymidine (DHdThd) residue—a damaged nucleoside generated from the thymidine (dThd) residue in DNA contained in food upon irradiation—was used as a detection indicator. Eight dried plant-based food samples were gamma ray-irradiated in the range from 3.2 to 8.3 kGy. Subsequently, DNA was extracted from the irradiated sample and digested into nucleosides by the three enzymes, and the test solution was analyzed by liquid chromatography tandem mass spectrometry (LC-MS/MS). Evidently, in all samples, the concentration ratio of DHdThd to dThd in the test solution (DHdThd/dThd) was dependent on the irradiation dose; moreover, during storage under frozen conditions for at least 890 d post-irradiation, this concentration ratio was equal to that immediately after irradiation. The irradiation histories of the eight types of dried plant-based food samples were correctly detected.

Chicken liver is a potential source of campylobacteriosis in humans. Therefore, we determined the number of Campylobacter in chicken liver. In total, 33 vacuum-packed liver products were obtained from retail stores, and found that 27 of the 33 products (81.8%) were contaminated with Campylobacter. Moreover, Campylobacter was isolated from 138 of 149 livers (92.6%) collected from the 27 Campylobacter-positive products. The mean Campylobacter count was 2.3 log₁₀ CFU/g, while Campylobacter count in 22 of the 138 contaminated livers (15.9%) was >3.0 log₁₀ CFU/g. Furthermore, gastrointestinal tract, liver, and bile samples were collected from 35 broilers at chicken processing plants. We isolated Campylobacter from the gastrointestinal tract of 27 broilers (77.1%). Of these 27 broilers, liver of 24 broilers (88.9%) was Campylobacter-positive, with a mean Campylobacter count of 2.8 log₁₀ CFU/g. Of these 24 broilers, bile of 13 broilers (54.2%) was contaminated with Campylobacter (mean Campylobacter count, 3.5 log₁₀ CFU/mL). Among them, bile of 2 broilers had a Campylobacter count of >8.3 log₁₀ CFU/mL. Collectively, these results indicate that livers derived from broilers colonized with Campylobacter are contaminated with Campylobacter at the time of evisceration. Therefore, to prevent foodborne campylobacteriosis in humans, chicken livers should be thoroughly heated before consumption.

Given that the number of genetically modified (GM) maize events that have been announced as having undergone safety assessment procedures in Japan is increasing yearly, more information is needed about their actual recent domestic distribution in Japan. In this study, we investigated whether current Japanese official qualitative and quantitative methods (the current official methods) for GM maize can comprehensively target events in domestically distributed maize. For samples with the identity-preserved (IP) handling system and non-IP samples from the United States (US) and non-IP samples from Brazil, we performed event-specific real-time PCR targeting 25 authorized single GM maize events in addition to the current official methods. According to our results, 15 events targeted by the current official methods were detected, but insect-resistance (IR) Event5307 and herbicide-tolerant (HT) DAS40278, not targeted by the current official methods, were detected in the US (one out of 5 lots) and Brazilian (four out of 5 lots) non-IP samples, respectively. Nevertheless, a survey of recent GM maize acreage in recent years has revealed that more than 95% of the acreage in US maize is occupied by HT or IR/HT stacked events, and that more than 95% of the acreage in Brazilian maize is occupied by IR or IR/HT stacked events. Because the current official methods can target all stacked events related to Event5307 and DAS40278, the only undetectable events are the single Event5307 and DAS40278, whose production is estimated to be less than 5% of the total production in the producing country. Therefore, we conclude that the current official methods for the labelling of GM maize should be maintained in view of practicability.

For the analysis of nitrite ions in food, the stabilities of nitrite ions in meat products and their standard solutions were evaluated. Nitrite is easily oxidized or reduced; hence, products with standard solutions or colour retention agent must be carefully handled. To assess the stability and decreasing trend of nitrite, we examined the storage stability of standard solutions using calibration curves, the time course of nitrite in chopped meat products stored under different conditions, and the time course of nitrite in the sample solutions. Regarding calibration curves, the storage stability was determined for standard solutions that were prepared with ultrapure water at concentrations of 0.025 and 0.4 μg/mL and were stored at 5℃ for one year. The results revealed no changes in concentration of any solution over time, suggesting that no readjustments to the standard solution concentration were necessary before testing until one year after their preparation. Time course of nitrite in chopped meat products stored under different conditions showed a significant decrease in nitrite in refrigerated storage (5℃), whereas stability of nitrite was maintained for up to 1 day in frozen storage (-20℃) and for 14 days in frozen storage (-40℃). The time course of nitrite in the sample solutions showed that the quantitative values of nitrite in the extract remained unchanged within one week of extraction for the meat products tested in the study.

Simple identification using a color reaction was applied to investigate poisoning, putatively caused by Omphalotus guepiniformis. Some leftover uncooked mushrooms had turned turquoise green when a beam reagent (5 w/v% potassium hydroxide ethanolic solution) was dripped onto the mushroom pileus. Furthermore, ethanol extract of the mushrooms exhibited the same color reaction. Then, illudin S, a toxic compound contained in O. guepiniformis, was detected in uncooked leftover mushrooms using LC-MS/MS analysis. Therefore, this case was inferred as caused by O. guepiniformis. These results indicate the identification method described above as useful for screening tests for investigating food poisoning caused by O. guepiniformis.

Severe tetrodotoxin (TTX) poisoning due to small gastropods has been documented in Japan. In this study, we investigated the TTX content of the muscles and viscera of Nassarius sufflatus collected off the coast of Futaoi Island, Shimonoseki, Yamaguchi Prefecture, Japan, to prevent the occurrence of TTX poisoning caused by this small gastropod. Live specimens were obtained, and their muscles and viscera were collected. Test solutions were prepared from tissues of specimens and analyzed for TTX by HPLC-fluorescence detection. TTX was detected in both tissues at concentrations ranging from <0.1 to 18.2 μg/g for muscle and <0.1 to 130.7 μg/g for viscera. These results suggested that N. sufflatus accumulates TTX not only in its viscera but also in its muscles, and that precautions should be taken to prevent food poisoning due to this gastropod.

In general, nitrite in food is extracted under slightly alkaline conditions, deproteinized, and analyzed by a colorimetric method using color development by diazotization. However, depending on the sample, the sample solution may become cloudy and difficult to filter by the deproteinization treatment of the analytical method. Recently, an improved analytical method that solves these problems has been reported. Therefore, a validation study was performed on the improved analytical method was performed. The concentrations of sodium nitrite added to cod roe, fish sausage, and ham, which were not labeled with sodium nitrite, were set at the upper limits of the standards for use. We set the target values of 70–120% for trueness, less than 15% for intralaboratory reproducibility, and less than intralaboratory reproducibility for repeatability. As a result, the target values were met for the three samples verified: 88–92% for trueness, 2.0–3.0% for repeatability, and 3.2–4.3% for intralaboratory reproducibility. In addition, an interlaboratory study was conducted by eight institutes on the improved analytical method for nitrite. At each institution, sodium nitrite was added to the same three samples as in the validation study, at concentrations equivalent to twice the lower limit of quantification and the upper limit of the standards for use and analyzed in triplicate. The estimated trueness from the obtained analyses ranged from 82 to 95%, the repeatability ranged from 2.3 to 5.8%, and the inter-room reproducibility ranged from 3.5 to 11%. Thus, the improved analytical method could be useful for determining nitrite in foods.

This study developed a method that simultaneously detected 283 pesticide residues in brown rice using GC-MS/MS and LC-MS/MS. In this method, we examined the desirable amount of sodium chloride required for salting out and the SPE cartridge required for clean-up. Pesticide residues from the sample were extracted with acetonitrile using a homogenizer and mixed with salts including anhydrous magnesium, two types of citrate, and sodium chloride. The sample solution of the acetonitrile layer was cleaned up using the GCB/NH₂ (200 mg/200 mg, 6 mL) SPE cartridge. The determination method was validated using two concentrations (0.01 and 0.1 μg/g) of 283 pesticides based on the validation guideline of the Ministry of Health, Labor and Welfare in Japan. Of the 283 pesticides, 250 were detected satisfactorily. In addition, 59 brown rice samples sold in Tokyo were surveyed using the same method. Out of 44 samples, 12 pesticide residues below MRLs were detected. Therefore, this developed method is useful for the simultaneous determination of pesticide residues in brown rice.