Food Hygiene and Safety Science最新文献

We assessed the applicability of high-performance liquid chromatography (HPLC) to the official testing method for the migration of caprolactam (CPL) from food utensils, containers, and packaging made from polyamide (PA). Hydrophilic interaction chromatography (HILIC) columns coated with unmodified silica, carbamoyl, and aminopropyl were used. Water and acetonitrile (ACN) were used as the mobile phase, and the analytical conditions were optimized. The test solution was diluted 10-fold with ACN, and standard solutions were prepared using 98% ACN. We validated using HPLC as limit testing and quantitative testing methods. Accuracy parameters corresponding to trueness, repeatability, and reproducibility (as intermediate precision) satisfied the target values in both cases, indicating that this method demonstrates good performance as a testing method.

A method was developed for determining five organochlorine unstable pesticides (captan, chlorothalonil, dichlofluanid, folpet, and tolylfluanid) in agricultural products using GC-MS/MS. To prevent pesticide degradation, the food sample was maintained at -20℃ until the initiation of the extraction process. The sample underwent homogenization with acetonitrile and salting out using anhydrous magnesium sulfate and sodium chloride in the presence of citrate salts. Cleanup was executed using Graphite Carbon and C18 SPE cartridges. The validation of this method was tested on five agricultural products at two concentrations (0.01 and 0.1 μg/g). The recovery rates varied between 86.7% and 96.1%. RSDs for repeatability were less than 15.2% and 8.8%, while the RSDs for within-laboratory reproducibility were under 19.9% and 12.7%. These results meet the criteria established by the validation guidelines in Japan.

In this study, a rapid and accurate analytical method was developed for the simultaneous determination of 26 plant toxins and 11 mushroom toxins in toxic plants, toxic mushrooms, and their cooked products using LC-MS/MS. This method enables highly selective detection of all 37 analytes, including those with high polarity and low molecular weight, within 10 min using Scherzo SS-C18 column. The analytes were extracted from the samples using methanol and trichloroacetic acid, and purified using Captiva EMR-Lipid. A 50 vol% methanol solution was used as the test solvent, to minimize matrix effects. Validation with six types of food samples demonstrated recoveries ranging from 56.0% to 180.5% (with 96% or more of analytes achieving 70-120%) and RSD of 16.0% or less (with 98% or more of analytes achieving 10% or less), at an added concentration of 1 mg/kg. These results indicate that this method is effective for health crisis management. Additionally, the method successfully detected expected toxins in food leftovers and reference products implicated in previous health hazard cases.

Since the establishment of procedures for the safety assessment of food products that use recombinant DNA technology, the manufacture, import, and sale of genetically modified (GM) foods that have not undergone safety assessment are prohibited under the Food Sanitation Act. Therefore, a performance study to confirm the GM food testing operations of each laboratory is very important to ensure the reliability of the GM food monitoring system. In 2022, GM papaya line PRSV-YK-which has not yet been authorized in Japan-was selected for testing, and a papaya paste and a DNA solution were used as the test samples. With these samples, a laboratory performance study of the DNA extraction and real-time PCR operations was conducted. This confirmed that the 18 participating laboratories were generally performing the DNA extraction and real-time PCR operations correctly. However, some laboratories using certain DNA amplification reagent with some real-time PCR instruments were not able to determine the PRSV-YK detection test. This suggests that the PRSV-YK detection test may not be able to correctly detect samples containing GM papaya when performed with these combinations of instruments and reagent. In order to ensure the reliability of the PRSV-YK detection test, it is necessary to examine in detail how the combination of DNA polymerase reagents and real-time PCR instruments affects the detection limit, and to implement an appropriate solution.

The present study verified that it is possible to analyze melengesterol acetate using the existing multi-residue method. Melengestrol acetate was extracted from livestock products using acidic acetonitrile acidified with acetic acid in the presence of n-hexane and anhydrous sodium sulfate. The crude extracts were cleaned up using an octadecylsilanized silica gel cartridge column. Separation by HPLC was performed using an octadecylsilanized silica gel column with linear gradient elution of 0.1 vol% formic acid and acetonitrile containing 0.1 vol% formic acid. For the determination of the analyte, tandem mass spectrometry with positive ion electrospray ionization was used. In recovery tests using four livestock products fortified with maximum residue limits levels of melengestrol acetate (0.001-0.02 mg/kg), the truenesses ranged from 82% to 100%, and the repeatabilities for the entire procedure ranged from 0.5 RSD% to 5.6 RSD%. In recovery tests using 11 livestock products fortified with 0.0005 mg/kg of melengestrol acetate, the truenesses ranged from 88% to 99%, and the repeatabilities ranged from 1.3 RSD% to 5.4 RSD%. The limit of quantification for melengestrol acetate in livestock products was 0.0005 mg/kg.

The upper and lower limits of microbial colony count on agar plate are essential for microbial cell calculation of food samples. In the present study, the limits presented by the three standards of Ministry of Health, Labor, and Welfare of Japan, Food and Drug Administration of US (FDA), and Japanese Industrial Standards (JIS) were studied from the stochastic point of view. Here colony counts per plate were assumed to follow the Poisson distribution from our previous studies. The probability of acceptance of a pair of colony counts, P_A for cell calculation by each standard was then calculated for various values of the mean of the Poisson distribution theoretically and by simulation with random sampling numbers. The values of P_A at the lower and upper limits of the standards were low. For example, P_A at the mean of 25 which is the lower limit of the FDA standard was 0.278. The values of P_A of colony count pairs by the three standards were demonstrated in a wide range of the mean colony count. The largest range of the mean at P_A>0.5 was observed in the JIS standard among the three. A new method to determine the limits for colony counts was developed with the risk of not acceptance. These findings would be helpful to consider the limits of colony counts for cell calculation.

Although the safety of genome-edited foods in Japan has been confirmed through pre-submission consultation under the notification process, public perception of safety confirmation methodology has not been investigated to date. Therefore, we created three media to provide information on the safety assurance of genome-edited foods and surveyed the perception of current safety confirmation. In addition, we examined the opinions of researchers in health science on current safety confirmation methods. As a result, 62% of general consumers and 68% of researchers in health science recognized that safety is ensured. Acceptance of genome-edited foods improved when they realized that safety was ensured. Researchers in health science who felt that safety confirmation was insufficient were concerned about the third-party verification. Therefore, it was suggested that in order to boost public understanding of genome-edited foods, it would be useful to inform the public by communicating in an easy-to-understand way the safety assurance approaches being made in pre-submission consultation.

We have developed a rapid genus identification method for poisonous plants. The real-time PCR using the TaqMan^® probe method was employed for detection, with the amplified targets being the "trnL (UAA)-intron" or "trnL-trnF intergenic spacer" regions of chloroplast DNA. The targeted plants were selected six genera (Aconitum, Colchicum, Veratrum, Brugmansia, Scopolia and Narcissus), which have been implicated in many instances of food poisoning in Japan. A tissue lysis solution was used for DNA extraction, which can be completed within approximate 30 min. A master mix corresponding to the tissue lysis solution was used for real-time PCR reagents. As a result, we were able to complete the entire process from DNA extraction to genus identification in 4 to 5 hr. The detection sensitivity was estimated at approximately 1 pg of DNA for all six plant genera. Remarkably, an amplification plot was discerned even with the crude cell lysates of all samples. It was also possible to obtain amplification curves for three plant samples that had been subjected to simulated cooking (boiling). This study suggests that the developed method can rapidly identify six genera of poisonous plants.

Nicotinic acetylcholine receptors (nAChRs) are neurotransmitter receptors found in the nervous system of many organisms, including humans. Neonicotinoid pesticides act as nAChRs modulators that affect neurotransmission. Due to toxicity effects, their use has been restricted. However, a new class of modulators (nAChRMs) have been developed, but analytical methods for the detection of residues of these new pesticides in agricultural crops have not been established. Therefore, this study aimed to develop and validate an accurate determination method for novel nAChRMs, such as sulfoxaflor, flupyradifurone, flupyrimin, and triflumezopyrim in brown rice using liquid chromatography with tandem mass spectrometry (LC-MS/MS). The method was applied to commercially available brown rice samples from Tokyo, Japan. Target analytes were extracted with acetonitrile, cleaned with GC/PSA, and then cleaned again with MonoSpin PBA. In accordance with the method validation guidelines for residual pesticides in foods, the performance characteristics were evaluated, with trueness ranging from 86.3% to 98.2%, repeatability of less than 6.5% relative standard deviation (RSD), and within-laboratory reproducibility of less than 6.5% RSD. These results indicate that the developed method can be applied to residue surveillance of target analytes using solvent standard calibration curves. By applying the developed method to 53 brown rice samples commercially available in Tokyo, sulfoxaflor residues were found in two samples at concentrations of 3.7 and 21.9 ng/g. This is the first report of the detection of sulfoxaflor residues in domestic agricultural products.

A method for analyzing tetrodotoxin (TTX) in miso soup samples was proposed. The samples were purified using strong cation exchange solid-phase extraction and analyzed by liquid chromatography-tandem mass spectrometry. The recovery of TTX was considerably influenced by the salt concentration in the loading solution during purification. It was observed that diluting the loading solution to reduce the salt concentration helped to maintain the recovery rate during the washing step. However, during the loading step, the benefit of dilution in improving recovery was not evident, as the enhanced retention on the solid phase caused by dilution was counteracted by losing TTX with the increased volume of the loading solution, which led to enhanced elution. This suggests that the volume of extraction solution used in the loading process is crucial in determining the recovery rate. Additionally, the use of volatile ammonium acetate as the elution solvent was explored to optimize conditions for effective recovery. Testing this method on miso soup samples with varying miso types, salt concentrations, and TTX levels resulted in consistently high recovery rates of>80%.