Development of a Reverse-Yield Factor Database Disaggregating Japanese Composite Foods into Raw Primary Commodity Ingredients Based on the Standard Tables of Food Composition in Japan.

The reverse-yield factor (RF) database was developed for qualitatively and quantitatively disaggregating Japanese composite foods into raw primary commodity (RPC) ingredients. Representative equations for four types (dried, salted, fermented and mixed foods) were developed to calculate RFs using the food content and composition data for composite foods listed in the Standard Tables of Food Composition in Japan-2020-(STFCJ), published by the Ministry of Education, Culture, Sports, Science and Technology of Japan. Out of 1150 composite foods identified in the STFCJ, RFs for 54 dried, 41 salted, 40 fermented and 818 mixed foods were obtained. RFs for 197 mixed foods could not be calculated because these foods were produced from ingredients with no specified information and/or through complex processing. The content and composition of Japanese composite foods would be interpreted representatively by RFs in the developed database.

Isolation of Shiga Toxin-Producing Escherichia coli from the Surfaces of Beef Carcasses in Slaughterhouses in Japan.

Shiga toxin-producing E. coli (STEC) is an important foodborne pathogen worldwide. It is necessary to control and prevent STEC contamination on beef carcasses in slaughterhouses because STEC infection is associated with beef consumption. However, the frequencies of STEC contamination of beef carcasses in various slaughterhouses in Japan are not well known. Herein, we investigated the contamination of beef carcasses with STEC in slaughterhouses to assess the potential risks of STEC. In total, 524 gauze samples were collected from the surfaces of beef carcasses at 12 domestic slaughterhouses from November 2020 to February 2023. The samples were measured for aerobic plate counts and tested for pathogenic genes (stx and eae) and major O-serogroups (O26, O45, O103, O111, O121, O145, and O157) by real-time PCR screening. Subsequently, immunomagnetic separation (IMS) was performed on samples positive for stx, eae, and at least one of the seven O-serogroups of STEC. Isolation process without IMS was performed on samples positive for stx, including those subjected to IMS. STEC O157:H7 and stx-positive E. coli other than serotype O157:H7 were isolated from 0.6% and 4.6% of beef carcass surfaces, respectively. Although the STEC O157:H7 isolation rate was low and stx-positive E. coli other than serotype O157:H7 belonged to minor O-serogroups, the results mean a risk of foodborne illness. Furthermore, a moderate correlation was observed between aerobic plate counts and detection rates of stx-positive samples by real-time PCR screening. The STEC O157:H7 isolated facilities showed higher values on aerobic plate counts and detection rates of stx-positive samples than the mean values of total samples. Therefore, these results suggest that it is important to evaluate hygiene treatments against beef carcasses for the reduction of STEC contamination risk, particularly in facilities with high aerobic plate counts.

[Analytical Method for Melengestrol Acetate in Livestock Products Using LC-MS/MS].

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.

[Implementation and Evaluation of Risk Communication Regarding Residual Pesticides].

In this study, a public seminar on risk communication methods was conducted to raise awareness and disseminate accurate knowledge about residual pesticides to consumers. Additionally, surveys on consumer awareness were conducted on the attendees before and after the seminar to evaluate its effectiveness. Responses were obtained from 84 participants. The paired t-test was used to analyze the changes in awareness before and after the seminar. The results showed significant improvements in "trust in the government" and "understanding of residual pesticides." Furthermore, step-wise multiple regression analysis was performed to explore the factors influencing satisfaction with the risk communication seminar, and the item "understanding of the safety of residual pesticides in food" was extracted. Understanding food safety is a crucial concern in daily life for consumers. To enable consumers to have an accurate understanding of food risks and make appropriate judgments, it is essential to continue implementing risk communication and conveying information about food safety and security in the future.

Development of a processing factor prediction model for pesticides in processed tomato foods using elastic net regularization.

A novel regularized elastic net regression model was developed to predict processing factor (PF) for pesticide residues, which represents a change in the residue levels during food processing. The PF values for tomato juice, wet pomace and dry pomace in the evaluations and reports published by the Joint FAO/WHO Meeting on Pesticide Residues significantly correlated with the physicochemical properties of pesticides, and subsequently the correlation was observed in the present tomato processing study. The elastic net regression model predicted the PF values using the physicochemical properties as predictor variables for both training and test data within a 2-fold range for 80-100% of the pesticides tested in the tomato processing study while overcoming multicollinearity. These results suggest that the PF values are predictable at a certain degree of accuracy from the unique sets of physicochemical properties of pesticides using the developed model based on a processing study with representative pesticides.

A reliable enzyme-linked immunosorbent assay for the determining of sesame proteins in raw food ingredients and in processed foods.

Sesame is a frequent cause of adverse food reactions in allergic patients. We developed a novel sandwich enzyme-linked immunosorbent assay (ELISA) using two monoclonal antibodies and a unique extraction buffer for the detection and quantification of sesame proteins in processed foods and in raw food ingredients to clarify the validity of sesame labeling and for precautionary allergen labeling. The developed sandwich ELISA method is highly specific for sesame proteins. The limit of detection (LOD) and limit of quantification (LOQ) are 0.013 µg/g and 0.025 µg/g, respectively. The recoveries for incurred food samples, such as dressing, breads, sauce and pudding, ranged from 67 % to 81 %, while the repeatability and reproducibility coefficients of variation were less than 4.7 % and 4.5 %, respectively. The developed method has applicability for food products and is a reliable tool for the detection of hidden sesame proteins in raw food ingredients and in processed foods.

Associations between cardiorespiratory fitness and lifestyle-related factors with DNA methylation-based ageing clocks in older men: WASEDA'S Health Study.

DNA methylation-based age estimators (DNAm ageing clocks) are currently one of the most promising biomarkers for predicting biological age. However, the relationships between cardiorespiratory fitness (CRF), measured directly by expiratory gas analysis, and DNAm ageing clocks are largely unknown. We investigated the relationships between CRF and the age-adjusted value from the residuals of the regression of DNAm ageing clock to chronological age (DNAmAgeAcceleration: DNAmAgeAccel) and attempted to determine the relative contribution of CRF to DNAmAgeAccel in the presence of other lifestyle factors. DNA samples from 144 Japanese men aged 65-72 years were used to appraise first- (i.e., DNAmHorvath and DNAmHannum) and second- (i.e., DNAmPhenoAge, DNAmGrimAge, and DNAmFitAge) generation DNAm ageing clocks. Various surveys and measurements were conducted, including physical fitness, body composition, blood biochemical parameters, nutrient intake, smoking, alcohol consumption, disease status, sleep status, and chronotype. Both oxygen uptake at ventilatory threshold (VO2 /kg at VT) and peak oxygen uptake (VO2 /kg at Peak) showed a significant negative correlation with GrimAgeAccel, even after adjustments for chronological age and smoking and drinking status. Notably, VO2 /kg at VT and VO2 /kg at Peak above the reference value were also associated with delayed GrimAgeAccel. Multiple regression analysis showed that calf circumference, serum triglyceride, carbohydrate intake, and smoking status, rather than CRF, contributed more to GrimAgeAccel and FitAgeAccel. In conclusion, although the contribution of CRF to GrimAgeAccel and FitAgeAccel is relatively low compared to lifestyle-related factors such as smoking, the results suggest that the maintenance of CRF is associated with delayed biological ageing in older men.

Technology Innovation and Guardrails in Elite Sport: The Future is Now.

A growing number of companies are developing or using wearable sensor technologies that can monitor, analyse and transmit data from humans in real time that can be used by the sporting, biomedical and media industries. To explore this phenomenon, we describe and review two high-profile sporting events where innovations in wearable technologies were trialled: the Tokyo 2020 Summer Olympic Games (Tokyo 2020, Japan) and the 2022 adidas Road to Records (Germany). These two major sporting events were the first time academic and industry partners came together to implement real-time wearable solutions during major competition, to protect the health of athletes competing in hot and humid environments, as well as to better understand how these metrics can be used moving forwards. Despite the undoubted benefits of such wearables, there are well-founded concerns regarding their use including: (1) limited evidence quantifying the potential beneficial effects of analysing specific parameters, (2) the quality of hardware and provided data, (3) information overload, (4) data security and (5) exaggerated marketing claims. Employment and sporting rules and regulations also need to evolve to facilitate the use of wearable devices. There is also the potential to obtain real-time data that will oblige medical personnel to make crucial decisions around whether their athletes should continue competing or withdraw for health reasons. To protect athletes, the urgent need is to overcome these ethical/data protection concerns and develop wearable technologies that are backed by quality science. The fields of sport and exercise science and medicine provide an excellent platform to understand the impact of wearable sensors on performance, wellness, health, and disease.

[Development of Analytical Method and Surveillance ofGibberellic Acid in Banana, Cherry, and Kiwi Fruit].

Gibberellic acid (GA3) is commonly used as a plant growth regulator in many food crops owing to its essential signaling functions during plant growth and development. In Japan, a threshold for administrative action for GA3 content of 0.3 mg/kg applies in produce in which maximum residue limits have not been established. Although the threshold is based on previous studies, the GA3 concentrations in individual foods are still unknown. Thus, we surveyed the concentrations of GA3 in banana, cherry, and kiwi fruit on the Japanese market. We developed and validated a method for the analysis of GA3 using solid-phase extraction and LC-MS/MS in accordance with accepted criteria of trueness, repeatability, and selectivity. The limits of detection and of quantification were determined as 0.005 and 0.05 mg/kg, respectively, in all fruits. Concentrations of GA3 did not exceed 0.3 mg/kg regardless of ripeness, suggesting the reasonability of the current regulation of GA3 in banana, cherry, and kiwi fruit. These findings can support prompt administrative action on these fruits, contributing to the regulation of GA3 in Japan.

Identification of animal species by nucleic acid chromatography of hair samples and investigation of its applicability to human biological samples.

GeneFields®-Hair is a simple analysis kit that uses nucleic acid chromatography and polymerase chain reaction (PCR) to identify animal species of hair-like food contaminants. In this study, we evaluated GeneFields®-Hair as a simple and rapid method for identifying animal species from hair-like materials collected in forensic science, such as at crime scenes. The use of this kit with other human biological materials (whole blood, head dandruff, nails, saliva, oral mucosa, sebum, and urine) was also investigated. Animal body hair samples were pretreated by grinding in a buffer solution, centrifuged, and the supernatant was used for PCR. Nucleic acid chromatography of the PCR products allowed the identification of the animal species by the presence or absence of coloration on the decision line. For human biological materials, nucleic acid chromatography was performed after the appropriate pretreatment like body hair material. The determination of some animal species was difficult, even if they had a dedicated DNA Strip determination line. Furthermore, animals from the same family but different genera were sometimes detected on the same determination line. All the human biological samples were correctly identified. Smartphone photographs of the coloration of the judgment line were processed using the ImageJ software for quantitative determination.

Estimated standard values of aerobic capacity according to sex and age in a Japanese population: A scoping review.

Aerobic capacity is a fitness measure reflecting the ability to sustain whole-body physical activity as fast and long as possible. Identifying the distribution of aerobic capacity in a population may help estimate their health status. This study aimed to estimate standard values of aerobic capacity (peak oxygen uptake [Formula: see text] and anaerobic threshold [AT]/kg) for the Japanese population stratified by sex and age using a meta-analysis. Moreover, the comparison of the estimated standard values of the Japanese with those of other populations was performed as a supplementary analysis. We systematically searched original articles on aerobic capacity in the Japanese population using PubMed, Ichushi-Web, and Google Scholar. We meta-analysed [Formula: see text] (total: 78,714, men: 54,614, women: 24,100) and AT (total: 4,042, men: 1,961, women: 2,081) data of healthy Japanese from 21 articles by sex and age. We also searched, collected and meta-analysed data from other populations. Means and 95% confidence intervals were calculated. The estimated standard values of [Formula: see text] (mL/kg/min) for Japanese men and women aged 4-9, 10-19, 20-29, 30-39, 40-49, 50-59, 60-69, and 70-79 years were 47.6, 51.2, 43.2, 37.2, 34.5, 31.7, 28.6, and 26.3, and 42.0, 43.2, 33.6, 30.6, 27.4, 25.6, 23.4, and 23.1, respectively. The AT/kg (mL/kg/min) for Japanese men and women aged 20-29, 30-39, 40-49, 50-59, 60-69, and 70-79 years were 21.1, 18.3, 16.8, 15.9, 15.8, and 15.2, and 17.4, 17.0, 15.7, 15.0, 14.5, and 14.2, respectively. Herein, we presented the estimated standard values of aerobic capacity according to sex and age in a Japanese population. In conclusion, aerobic capacity declines with ageing after 20-29 years of age. Additionally, aerobic capacity is lower in the Japanese population than in other populations across a wide range of age groups. Standard value estimation by meta-analysis can be conducted in any country or region and for public health purposes.

Optimization of QuEChERS Extraction for Determination of Carotenoids, Polyphenols, and Sterols in Orange Juice Using Design of Experiments and Response Surface Methodology.

Several compounds with different physical properties are present in foods, biological components, and environmental samples, and there are cases in which these must be analyzed simultaneously. However, it is difficult to extract compounds with different physical properties from the same sample using a single method. In the present study, we examined the optimal conditions for the QuEChERS extraction of several kinds of compounds from orange juice using design of experiments (DoE) and response surface methodology (RSM) to determine the optimal ratio of organic solvent to sodium chloride. We determined the optimal extraction conditions, which were within the design space, using 100% tetrahydrofuran (THF) as the extraction organic solvent and NaCl:MgSO4 = 75:25 as the salt. The developed LC/MS/MS method using QuEChERS extraction achieved specific detection and precise quantification. Finally, we measured the polyphenols, sterols, and carotenoids in citrus juice using the optimized QuEChERS extraction method before LC/MS/MS analysis. Most of the analytes were quantifiable in orange juice. The optimized method achieved ease of operation, the extraction of analytes from food samples in a short time (within 30 min), minimization of analytical residues, and reliability. The DoE and RSM approach may contribute to better optimization of the extraction conditions for the lowest number of experiments.

Determination of Cyanide and Cyanoglycosides in Sweetened Bean Paste by HPLC with Fluorescence Detection.

It is necessary to evaluate the efficiency of reduction for cyanide and cyanoglycosides during the manufacturing process from raw material beans to sweetened bean paste in a food hygiene control system from the viewpoint of food safety. Analytical methods for cyanide and cyanoglycoside determination in sweetened bean paste by HPLC with fluorescence detection were developed. In analysis of collection time of free cyanide in the free cyanide assay, the recovery was improved by extending the collection time, the recovery rate was >80% by 2 h. The accuracy, repeatability and intra-laboratory precision of the free cyanide assay were 82.3, 2.0, and 2.4%, respectively. The method for cyanoglycoside analysis was evaluated by 5 repeated spiked recovery experiments at a concentration of 10 ppm. The accuracy, repeatability and intra-laboratory precision of the cyanoglycoside method were 82.2, 1.9, and 3.4%, respectively. These analytical methods will enable the analysis of cyanide and cyanoglycosides in sweetened bean paste without using steam distillation method in the pretreatment.

Acute social jetlag augments morning blood pressure surge: a randomized crossover trial.

Although social jetlag (SJL) is generally considered a chronic condition, even acute SJL may have unfavorable effects on the cardiovascular system. We focused on the acute effects of SJL on morning blood pressure (BP) surge. This randomized crossover trial recruited 20 healthy men. In the SJL trial, participants delayed their bedtime by three hours on Friday and Saturday nights. Participants in the control (CON) trial implemented the same sleep-wake timing as on weekdays. Pre- and post-intervention measurements were performed to evaluate resting cardiovascular variables on Friday and Monday mornings, respectively. The ambulatory BP was automatically measured during the sleep and awake periods for 2 h after the participant woke up at night before pre- and post-intervention measurements. SJL (average mid-sleep time on weekends - average mid-sleep time on weekdays) occurred only in the SJL trial (SJL: 181 ± 24 min vs. CON: 8 ± 47 min). Carotid-femoral pulse wave velocity (cfPWV) and morning BP surge on Monday in the SJL trial were significantly higher than those on Friday in the SJL trial (cfPWV: P = 0.001, morning BP surge: P < 0.001), and those on Monday in the CON trial (cfPWV: P = 0.007; morning BP surge: P < 0.001). Furthermore, a significant positive correlation was found between ΔcfPWV and Δmorning BP surge (R = 0.587, P = 0.004). These results suggest that even acute SJL augments morning BP surge. This phenomenon may correspond to increased central arterial stiffness.State the details of Clinical Trials: Name: Effect of acute social jetlag on risk factors of lifestyle-related diseases. URL: https://center6.umin.ac.jp/cgi-open-bin/ctr_e/ctr_view.cgi?recptno=R000053204 . Unique identifier: UMIN000046639. Registration date: 17/01/2022.

[An LC-MS/MS Analytical Method for Moenomycin A in Livestock Products].

A simple and sensitive method for the determination of moenomycin A residues in livestock products using LC-MS/MS was developed. Moenomycin A, a residual definition of flavophospholipol, was extracted from samples with a mixture of ammonium hydroxide and methanol (1 : 9, v/v) preheated at 50℃. The crude extracted solutions were evaporated and purified by liquid-liquid partitioning between a mixture of ammonium hydroxide, methanol and water (1 : 60 : 40, v/v/v) and ethyl acetate. The alkaline layer was taken, and cleaned up using a strong anion exchange (InertSep SAX) solid phase extraction cartridge. The LC separation was performed on an Inertsil C8 column with liner gradient elution using 0.3 vol% formic acid and acetonitrile containing 0.3 vol% formic acid. Moenomycin A was detected using tandem mass spectrometry with negative ion electrospray ionization. Recovery tests were conducted using three porcine samples (muscle, fat and liver) and chicken eggs. Samples were spiked with moenomycin A at 0.01 mg/kg and at the Japanese Maximum Residue Limits (MRLs) established for each sample. The trueness ranged from 79 to 93% and precision ranged from 0.5 to 2.8%. The limit of quantification (S/N≥10) of the developed method is 0.01 mg/kg. The developed method would thus be very useful for regulatory monitoring of flavophospholipol in livestock products.

Elucidation of Degradation Behavior of Hydroxy Group-Containing Benzodiazepines in Artificial Gastric Juice: Study on Degradability of Drugs in Stomach (III).

The degradation behavior of three benzodiazepines (BZPs)-lormetazepam (LMZ), lorazepam, and oxazepam-with hydroxy groups on the diazepine ring in artificial gastric juice and the effect of storage pH conditions on drug degradability were monitored using an LC/photodiode array detector (PDA) to estimate their pharmacokinetics in the stomach. Although the three BZPs degraded in artificial gastric juice, none could be restored, despite increasing the storage pH, implying that the degradation reaction was irreversible. As for LMZ, we discussed the physicochemical parameters, such as the activation energy and activation entropy involved in the degradation reaction as well as the reaction kinetics; one of the degradation products was isolated and purified for structural analysis. In the LMZ degradation experiment, peaks corresponding to degradation products, (A) and (B), were detected through the LC/PDA measurements. Regarding the degradation behavior, we hypothesized that LMZ was degraded into (B) via (A), where (A) was an intermediate and (B) was the final product. Although the isolation of degradation product (A) was challenging, degradation product (B) could be isolated and was confirmed to be "methanone, [5-chloro-2-(methylamino)phenyl](2-chlorophenyl)-" based on structure determination using various instrumental analyses. The compound exhibited axis asymmetry as determined using single-crystal X-ray structure analysis. Because the formation of degradation product (B) was irreversible, it would be prudent to target the final degradation product (B) and LMZ for identification when detecting LMZ in human stomach contents, such as during forensic dissection.

[Estimation of the Distribution of Dietary Exposure to Lead Using Two-Dimensional Monte Carlo Simulation: An Attempt to Apply to Total Diet Samples].

The average dietary exposure to lead (Pb) in male and female Japanese individuals >1 year of age was estimated using 280 total diet samples representing 14 food groups from 10 areas over a two-year period. A probabilistic exposure estimation was performed using a two-dimensional Monte Carlo simulation (2D-MCS) with a Bayesian estimation that consided the uncertainty of the estimation process. The Bayesian estimation was performed using the likelihood function with cumulative distribution function between the lower and upper boundary values for no-detected values. The median dietary exposure to Pb was estimated as 5.82 µg/person/day. The 90% interval was 2.51-16.9 µg/person/day. Comparison with previously reported Pb exposure values indicates that the estimation of Pb exposure distribution using total diet samples is reasonable. The contribution to Pb exposure was highest in the order of food group 8 (light-colored vegetables, mushrooms, and seaweeds: 20.0±16.1%)>food group 1 (rice and rice products: 12.3±19.0%)>food group 10 (fish and shellfish: 10.5±13.9%). Owing to the high uncertainties of contribution ratios, it was not possible to identify dominant food groups contributing to Pb exposure. However, it was evident that the uncertainty of the estimation of Pb exposure was influenced by the uncertainty of Pb concentration than the uncertainty of food consumption rate. In particular, the effect of uncertainty from the Pb concentration of the food group 1 was 68.2%. When the margin of exposures were calculated, the estimated probabilities that a value would be <1 were 14.5% for developmental neurotoxicity to children (1-6 years old), 0.13% for blood pressure and 0.93% for kidney disease in Japanese individuals ≥1 year of age. The findings suggest that the health risk due to dietary Pb exposure is small but not negligible.

Simultaneous determination of glyphosate, glufosinate, and their metabolites in honey using liquid chromatography-tandem mass spectrometry and solid-phase extraction.

Here, we developed and validated a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the simultaneous determination of residual glyphosate, glufosinate, and their metabolites N-acetylglyphosate (Gly-A), 3-methylphosphinicopropionic acid (MPPA) and N-acetylglufosinate (Glu-A) in honey using a mixed mode column of reversed-phase and anion exchange without derivatization. The target analytes were extracted from honey samples using water, cleaned up on a reverse phase C18 cartridge column and an anion exchange NH2 cartridge column, and quantified using LC-MS/MS. Glyphosate, Glu-A, Gly-A, and MPPA were detected in negative ion mode based on deprotonation, whereas glufosinate was detected in positive ion mode. The coefficients of determination (R2) of the calibration curve, calculated in the range of 1-20 µg/kg for glufosinate, Glu-A, and MPPA, and 5-100 µg/kg for glyphosate and Gly-A, were higher than 0.993. The developed method was evaluated using honey samples spiked with glyphosate and Gly-A at 25 µg/kg and glufosinate, and MPPA and Glu-A at 5 µg/kg, based on the maximum residue levels. The validation results show good recoveries (86-106%) and precision (< 10%) for all target compounds. The limit of quantification of the developed method is 5 µg/kg for glyphosate, 2 µg/kg for Gly-A, and 1 µg/kg for glufosinate, MPPA and Glu-A. These results suggest that the developed method is applicable for quantifying residual glyphosate, glufosinate, and their metabolites in honey in compliance with Japanese maximum residue levels. Moreover, the proposed method was applied to the analysis of honey samples and glyphosate, glufosinate, and Glu-A were detected in some samples. The proposed method will be a useful tool for the regulatory monitoring of residual glyphosate, glufosinate, and their metabolites in honey.

Development of a simple and reliable LC-MS/MS method to simultaneously detect walnut and almond as specified in food allergen labelling regulations in processed foods.

We developed a simple and reliable analytical method using high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) to simultaneously detect walnut and almond as specified in regulations for food allergen labelling in processed foods. Five specific target peptides derived from walnut 2S albumin and 7S globulin and three target peptides from almond 11S globulin were selected by analysing several varieties of walnut and almond, eight kinds of other nuts, and ten kinds of major allergen ingredients or cereals. The limit of detection for the walnut 2S albumin peptide GEEMEEMVQSAR (m/z 698.3 [precursor] > 316.1 [product]) was 0.22 ± 0.02 µg/g, and that for almond 11S globulin peptide GNLDFVQPPR (m/z 571.8 [precursor] > 369.2 [product]) was 0.08 ± 0.02 µg/g when extracted walnut and almond protein were spiked into butter cookie chocolate ice cream. These peptides had good linearity (R2 > 0.999) for each calibration curve with a range of 0.1-50 µg/mL protein concentration in the sample solutions, and sufficient recovery rates (90.4-101.5%) from the spiked samples. The developed analytical approach is applicable to a wide variety of processed foods for food allergen labelling.

Development of the ultra-weak chemiluminescence method based on luminol reaction for use in the detection of ultra-trace levels of blood.

In this study, we focused on ultra-weak chemiluminescence (uwCL) measurements and aimed to develop a blood detection method at trace levels, that are difficult to observe using a conventional luminol reaction method (visual observation). Furthermore, we investigated sampling methods that could detect trace bloodstains from the field in our laboratory settings. To achieve these highly sensitive detection levels in the uwCL measurements, the optimal measurement conditions were established as follows: the luminol reaction solution chosen was a mixture of 6.0 × 10-3% luminol/1.5 × 10-2% sodium hydroxide solution and 1.5 × 10-2% hydrogen peroxide water (6:1); the temperature in the sample chamber was set to 20 °C; the sample chamber was filled with an oxygen displacement atmosphere; the sample chamber was filled with 3 mL of 0.01% sodium hydroxide solution prior to the experiment, and the measurement wavelength was set to 460 nm. Using the developed method, blood diluted to 12.5 million-fold (8.0 ng equivalent of the absolute weight of whole blood) was detectable, and high linearity (r = 0.9986) between uwCL intensity and whole blood equivalent was observed in the range of 8.0-100 ng. In contrast, the detection limit of the conventional method was 1.0 µg of the whole blood equivalent. Thus, the uwCL method was approximately 125 times more sensitive than the conventional method. In addition, we demonstrated that the sampling method of wiping with a melamine sponge soaked in a 10% sodium dodecyl sulfate solution is effective for sampling evidence materials at an appraisal site suspected of having traces of adhered blood.