The distribution of livestock products containing veterinary drugs exceeding the residue standards established under the Food Sanitation Act is prohibited. The freshness and quality of livestock products deteriorate over time, necessitating rapid and highly precise inspection methods. In this study, a multiresidue method for determination of veterinary drugs in pig muscle was developed, with a focus on tetracycline antibiotics, which account for a large proportion of total veterinary drug sales. To achieve rapid analysis, the QuEChERS method was employed as the basis for extraction, and membrane filtration (SPEEDIA) was used for sample cleanup. The extraction conditions (solvent type, chelating agent concentration, and pH) and the purification process were optimized. As a result, the trueness of the four tetracycline antibiotics (n=12) ranged from 97 to 111% at the spiking level of 0.01 mg/kg and from 84 to 111% at 0.1 mg/kg. In addition, it was confirmed that approximately 90% of all compounds in pig muscle samples satisfied the target criteria for recovery and precision specified in the method validation guidelines. Furthermore, the comparison with the official method demonstrated that sample pretreatment time and organic solvent consumption could be significantly reduced. These findings suggest that the proposed method is a useful, rapid, and simple analytical approach for the simultaneous determination of multiple veterinary drugs in pig muscle.

A simultaneous analytical method was developed for the determination of deoxynivalenol (DON) and ochratoxin A (OTA) in wheat. Analytes were purified from wheat extracts by a multifunctional column and quantified by LC-MS/MS. An interlaboratory study was conducted to validate the analytical method using three spiked and two contaminated wheat samples. The ranges of mean recoveries of DON and OTA were 88–89% and 91–96%, respectively. The RSDs for repeatability and reproducibility were all ≤15%. Furthermore, the levels of DON and OTA in artificially contaminated wheat samples were analyzed by the simultaneous analytical method. The results showed that the developed analytical method yielded analytical values equivalent to those obtained when analyzing DON and OTA individually by existing analytical methods. Based on these results, the simultaneous analytical method for DON and OTA in wheat developed in this study is considered suitable as an alternative to the methods for analyzing each mycotoxin individually.

This study compared microbial detection among game meat samples processed under different conditions. Game animals captured in Oita and Osaka Prefectures between November 2019 and March 2023 were classified into three groups according to processing conditions: on-site processing (n=8), field-based processing (n=30), and licensed facility processing group (n=223). Swab samples were collected from the abdominal cavity and thigh muscle, and aerobic bacteria, coliform bacteria, and staphylococci were examined using commercially available stamp agar plates. Detection rates were compared using Fisher’s exact test.

Bacteria were detected in all individuals processed under outdoor conditions (on-site and field-base groups), whereas no bacteria were detected in the licensed facility processing group (0/223)(p<0.001). No significant differences were observed between the two outdoor processing groups.

These findings indicate that the on-site and field-based processing groups showed higher detection rates of indicator bacteria than the licensed facility processing group. However, this was an observational study, and the findings do not directly demonstrate a causal effect of processing conditions. The results may also have been affected by regional differences, capture methods, seasonal differences, limited sample size in the on-site group, and pre-selection before facility entry.

Ethylene oxide (ETO) is a highly reactive sterilizing agent classified as a carcinogen, and its use in foods is prohibited in many countries. Regulatory monitoring currently relies primarily on the detection of ETO and/or 2-chloroethanol (2CE), a chloride-derived reaction product. However, ETO is volatile and difficult to detect directly in commercial foods, and 2CE may occur naturally or arise from non-food-related sources, raising scientific concerns regarding its specificity as a fumigation marker. In this study, we applied a food-processing metabolomics approach to comprehensively investigate the chemical alterations induced by ETO fumigation of cumin (Cuminum cyminum L.) seeds using gas chromatography-mass spectrometry. Fumigated and non-fumigated samples were clearly discriminated by principal component analysis of the data containing fumigation-specific metabolites. Here, we report that high-resolution gas chromatography quadrupole time-of-flight mass spectrometric analysis identified a characteristic compound with a molecular formula consistent with a trimethylsilylated derivative of C₆H₁₀O₆ containing a single ETO incorporation. Fragmentation patterns and accurate mass measurements suggested that this compound was an ETO adduct of malic acid. In vitro fumigation experiments using authentic malic acid reproduced the same chromatographic retention time and mass spectrum pattern, confirming its formation. A targeted gas chromatography-tandem mass spectrometric analysis further demonstrated the presence of the malic acid-ETO adduct (m/z 233 → 73) in fumigated sesame and soybean, indicating its potential broader presence across ETO fumigated plant-derived matrices. Collectively, these findings provide the first direct evidence that ETO fumigation induces the formation of a nutrient-derived adduct in foods. The identified ETO-modified nutrient represents a promising chemical marker for verifying ETO fumigation beyond conventional 2CE-based approaches, thereby contributing to improved analytical specificity and regulatory reliability in food safety control.

A reliable and sensitive analytical method was developed for the determination of stilbenes—namely diethylstilbestrol, dienestrol, hexestrol, and their conjugates—in bovine muscle. The method involves the addition of internal standards to samples, extraction with ethanol/water (9 : 1, v/v),enzymatic hydrolysis using β-glucuronidase and aryl sulfatase, cleanup using hydrophilic-modified styrene–divinylbenzene copolymer and aminopropyl silica solid-phase extraction cartridges, and subsequent LC-MS/MS analysis. The method was validated for diethylstilbestrol, dienestrol, and hexestrol in bovine muscle spiked at 0.5 μg/kg for each analyte. The results demonstrated satisfactory analytical performance, with trueness values of 94–100%, repeatability below 5%, and within-laboratory reproducibility below 8%. No interfering peaks were observed, confirming the high selectivity of the method, and matrix effects were negligible. The developed method is applicable not only to diethylstilbestrol and its conjugates regulated in Japan but also to dienestrol and hexestrol, whose use for growth promotion in food-producing animals is prohibited in the EU. Although the conjugates of dienestrol and hexestrol were not directly validated in this study, they were presumed to be quantifiable based on their structural similarity to diethylstilbestrol conjugates.

After addressing analytical issues, including the decomposition of diphenyl carbonate (DPC), we developed a modified analytical method for the determination of DPC and bisphenol A (BPA) in polycarbonate food apparatuses, containers, and packaging. In this method, the specification value for BPA was defined as the sum of BPA, phenol (PH), and p-tert-butylphenol (BuPH). Following the improvement of the analytical procedure, a validation study was conducted to evaluate the performance of the modified analytical method. The repeatability, within-laboratory reproducibility, and trueness were estimated to be 0.1–0.3%, 0.3–0.9%, and 98–100% for DPC; 0.2–0.3%, 0.3–1.5%, and 101–102% for BPA; 0.2–0.4%, 0.4–0.7%, and 97–98% for PH; and 0.2–0.4%, 0.2–0.4%, and 99–100% for BuPH, respectively, at three concentration levels. These results indicate that the modified analytical method is suitable for evaluating compliance with the specifications for DPC and BPA.

In response to the enforcement of the positive list (PL) system for food containers and packaging, we investigated the applicability of pyrolysis GC-MS as a testing method for evaluating compliance with this list. Samples were heated using a pyrolysis device, and the analytical conditions were established based on thermograms obtained from evolved gas analysis (EGA)-MS. Volatile components, such as additives and residual monomers, were detected using thermal desorption (TD)-GC/MS, whereas polymeric components and other base materials were analyzed using pyrolysis (Py)-GC/MS. The results demonstrated that the method enabled the estimation of the compounds and materials. However, similarly to previously reported methods aimed at compliance with the PL system, compounds that did not match the library or those presumed to be non-intentionally added substances (NIAS) were also detected. However, compared to conventional methods, the proposed approach requires a smaller sample amount, does not need pretreatment, is simple and rapid, allows material identification in a single series of analyses, and can detect additives at low levels. Therefore, this method is considered useful for monitoring and providing guidance related to the PL system.

Gloves that come into contact with food are required to comply with the Japanese Food Sanitation Act. However, it has been reported that licensed food establishments do not always use gloves that meet these requirements. In this study, evaporation residue tests were conducted on polyvinyl chloride gloves, and qualitative analysis of migrants was performed to assess compliance with the positive list. Furthermore, the contents and migration levels of six phthalates were quantified. While samples labeled as compliant with the Act met the standards, all samples that were either non-labeled or of unknown compliance exceeded the evaporation residue standard when heptane was used as a food-simulating solvent. Analysis of the major migrant substances revealed that some components in two samples were not included on the positive list. Moreover, quantification of six phthalates revealed that one sample contained bis (2-ethylhexyl) phthalate. The migration level of this sample was approximately 3000 µg/mL. These findings indicate the need for enhanced guidance to licensed food establishments to promote the use of compliant gloves, as well as increased awareness among general consumers.