Some rice bran pickles contain histamine (Him) and tyramine (Tym), which are nonvolatile amines, presumably produced by microorganisms during the fermentation process. When attempting to identify the histamine- and tyramine-producing bacteria from commercially available cucumber rice bran pickles containing Him and Tym, it was identified histamine-producing bacteria were found to be Raoultella ornithinolytica, and tyramine-producing bacteria were Lactobacillus curvatus. When those bacteria were cultured in media containing rice bran (the raw material for rice bran pickles), Him and Tym were produced. However, it was suggested that Him and Tym were not necessarily produced by the presence or absence of amine-producing bacteria, but rather, their production was affected by the amount of precursor amino acids and other bacteria that were present during rice bran pickles production.

Total diet study (TDS) is a useful way to estimate the dietary intake of hazardous and chemical substances. Regarding the analysis performed in TDS, international guidelines published by the World Health Organization, recommend selecting and confirming the validity of suitable analytical methods to achieve the purpose of TDS. However, concrete procedures and/or approaches for confirming the validity of suitable methods have yet to be established. In the present study, we aimed to develop samples, referred to as SEMPs; Samples to estimate methods performance, that can be used to evaluate the performance of the analytical methods applied to the composite samples prepared in TDS. The concentrations of 14 kinds of elements, including hazardous substances such as arsenic, lead, and cadmium, in SEMPs were measured for use in the validation of a multi-element analytical method for estimating dietary intake. After examining the appropriate amount of relevant elements added to the samples, we established a performance evaluation procedure by repeatedly analyzing five fortified and non-fortified SEMPs each.

Oranges are consumed worldwide; however, they contain Cit s 2, a major profilin allergen. We aimed to reduce Cit s 2 levels by preparing mixed orange fresh juice with pineapple, as a convenient method for any kitchen. Cit s 2 levels in orange extracts digested with pineapple extract and its protease bromelain were evaluated with quantitative enzyme-linked immunosorbent assay. Cit s 2 levels decreased according to reaction temperature and time, which was inhibited by iodoacetic acid. Treatment with pineapple extract diluted 40-fold and 0.1 mg/mL of bromelain at 37℃ for 30 min contributed to reducing residual Cit s 2 levels below the cut-off of 15%, respectively. Since this condition can increase the proportion of orange juice and reduce the risk of ingesting the pineapple allergen bromelain, it is considered to be more practical. Broad utilization of proteases in hypoallergenic food products is expected following clinical studies for verification.

An identification method for testing contamination in products was assessed using various vegetables and fruits (70 types in total). DNA was extracted from plant fragments which are 1 to several millimeters long and the plastid rpl16–rpl14 linker sequence (approximately 550 base pairs) was amplified by PCR. The DNA nucleotide sequence was determined, and homology and SNP (single nucleotide polymorphism) analyses were carried out. Consequently, the test plants were difficult to distinguish between closely related species, but could be divided into 38 groups at the genus level or the species level. Although problems such as the accuracy of discrimination among some closely related plants and DNA stability under an acidic condition remain to be resolved, this method is considered to be expected to identify plant fragments mixed in products or raw materials.

The mouse bioassay for tetrodotoxin has been used in Japan and ddY strain mice are designated to be used in the assay. ddY strain is a closed-colony outbred mouse strain, originally from the National Institute of Health, however, the ddY mouse stocks of 3 breeders were divided 30 or more years ago. In this study, we investigated the differences in susceptibility to tetrodotoxin in ddY strain mice from 3 different breeders in Japan. No statistically significant differences were found among the ddY strain mice of 3 breeders, however, the coefficient of variation were different among the breeders. The results using the mice from one breeder were much stable compared with those using the mice from other 2 breeders.

Some illegal dietary supplements contain phosphodiesterase type 5 (PDE5) inhibitors, such as sildenafil, for exerting “therapeutic” effects in erectile dysfunction. This is apparently dangerous, and thus, should be appropriately regulated. Identification of descarbonsildenafil was first reported in Singapore in a coffee sample labeled to exert male sexual performance enhancement effects. However, it is unclear whether the compound possesses PDE5 inhibitory activity. We encountered during our survey of dietary supplements, a sexual enhancement product commercially available in Tokyo, in which a peak presumed to be of descarbonsildenafil was detected by LC-UV and electrospray ionization-tandem MS (ESI-MS/MS). The compound was isolated and identified as descarbonsildenafil with liquid chromatography-quadrupole time-of-flight-mass spectrometry (LC-QTOF-MS), NMR, and X-ray crystal structural analysis. In addition, descarbonsildenafil showed PDE5 inhibitory activity in PDE5 inhibition assay, and its IC₅₀ value for PDE5A1 was found to be 30 nmol/L. The results of INADEQUATE NMR and X-ray crystal structural analysis in this study provide information for the identification of descarbonsildenafil. Since this study indicates that this compound is a PDE5 inhibitor having adequate activity, it is regulated as a drug component in Japan.

DNA from hair that has undergone retort pouch sterilization shows considerably more fragmentation. Assessing the extent of DNA fragmentation using a PCR assay could therefore be applied to infer the sterilization history of a contaminated food sample. Such assessments could make it possible to determine whether a food sample had been contaminated by hair during the production process. To determine the extent of retort pouch sterilization, primers specific for the detection of human DNA were designed to give an amplification product of approx 500 bp. Hair was subjected to retort sterilization as a model of contamination, and PCR was performed using the extracted DNA as a template and the primer set for determining retort pouch sterilization. The results showed that no DNA amplification occurred in retort pouch samples, whereas amplification was observed in samples that were unheated, heated in hot water, or heated in a microwave oven. The present method was thus able to specifically detect thermal decomposition of DNA from hair that had undergone retort sterilization, and will be useful for determining whether hair discovered in a retort pouch was responsible for contamination.

As a method to detect pesticide residues in agricultural products, we evaluated the pretreatment of agricultural product samples by the solid-phase extracton technique with QuEChERS (STQ) method followed by GC-MS/MS with large-volume injection using a stomach-type glass-lined injector. This method satisfied the target criteria of recovery (70–120%) and the standard deviation of repeatability (RSD<25%) in 238–282 pesticides found in six types of agricultural products.