Validation of analytical method for furan determination in eight food matrices and its levels in various foods.

In this study, an automated solid-phase microextraction coupled with gas chromatography and mass spectrometry method was developed and validated for the determination of furan in eight matrices including ham, milk, apple juice, rice porridge, peanut butter, flatfish, tuna (canned) and seaweed. The calibration curves were highly linear (r2  > 0.990) and the limit of detection and limit of quantification ranged from 0.01-0.02 and 0.04-0.06 ng/g, respectively. The recovery ranged from 77.81-111.47%. The validated method was used to analyse the furan levels in 120 foods. The highest levels of furan were detected in black tea (172.05 ng/g) and red ginseng extract (89.27 ng/g). Whelk (canned) contained a high furan content (21.34 ng/g) among the seafood samples.

Determination of glyoxal, methylglyoxal, and diacetyl in red ginseng products using dispersive liquid-liquid microextraction coupled with GC-MS.

A simple and rapid dispersive liquid-liquid microextraction method coupled with gas chromatography and mass spectrometry was applied for the determination of glyoxal as quinoxaline, methylglyoxal as 2-methylquinoxaline, and diacetyl as 2,3-dimethylquinoxaline in red ginseng products. The performance of the proposed method was evaluated under optimum extraction conditions (extraction solvent: chloroform 100 µL, disperser solvent: methanol 200 µL, derivatizing agent concentration: 5 g/L, reaction time: 1 h, and no addition of salt). The limit of detection and limit of quantitation were 1.30 and 4.33 µg/L for glyoxal, 1.86 and 6.20 µg/L for methylglyoxal, and 1.45 and 4.82 µg/L for diacetyl. The intra- and interday relative standard deviations were <4.95 and 5.80%, respectively. The relative recoveries were 92.4-103.9% in red ginseng concentrate and 99.4-110.7% in juice samples. Red ginseng concentrates were found to contain 191-4274 µg/kg of glyoxal, 1336-4798 µg/kg of methylglyoxal, and 0-830 µg/kg of diacetyl, whereas for red ginseng juices, the respective concentrations were 72-865, 69-3613, and 6-344 µg/L.

Solid-phase microextraction Arrow for the volatile organic compounds in soy sauce.

A novel solid-phase microextraction Arrow was used to separate volatile organic compounds from soy sauce, and the results were verified by using gas chromatography with mass spectrometry. Solid-phase microextraction Arrow was optimized in terms of three extraction conditions: type of fiber used (polydimethylsiloxane, polyacrylate, carbon wide range/polydimethylsiloxane, and divinylbenzene/polydimethylsiloxane), extraction temperature (40, 50, and 60°C), and extraction time (10, 30, and 60 min). The optimal solid-phase microextraction Arrow conditions were as follows: type of fiber = polyacrylate, extraction time = 60 min, and extraction temperature = 50°C. Under the optimized conditions, the solid-phase microextraction Arrow was compared with conventional solid-phase microextraction to determine extraction yields. The solid-phase microextraction Arrow yielded 6-42-fold higher levels than in solid-phase microextraction for all 21 volatile organic compounds detected in soy sauce due to the larger sorption phase volume. The findings of this study can provide practical guidelines for solid-phase microextraction Arrow applications in food matrixes by providing analytical methods for volatile organic compounds.

Dispersive liquid-liquid microextraction with in situ derivatization coupled with gas chromatography and mass spectrometry for the determination of 4-methylimidazole in red ginseng products containing caramel colors.

A rapid analytical method was developed for the determination of 4-methylimidazole from red ginseng products containing caramel colors by using dispersive liquid-liquid microextraction with in situ derivatization followed by gas chromatography with mass spectrometry. Chloroform and acetonitrile were selected as the extraction and dispersive solvents, and based on the extraction efficiency, their optimum volumes were 200 and 100 µL, respectively. The optimum volumes of the derivatizing agent (isobutyl chloroformate) and catalyst (pyridine), pH, and concentration of NaCl in the sample solution were determined to be 25 and 100 µL, pH 7.6, and 0% w/v, respectively. Validation of the optimized method showed good linearity (R2  > 0.999), accuracy (≥89.86%), intra- (≤6.70%) and interday (≤4.17%) repeatability, limit of detection (0.96 µg/L), and limit of quantification (5.79 µg/L). The validated method was applied to quantify 4-methylimidazole in red ginseng juices and concentrates, 4-methylimidazole was only found in red ginseng juices containing caramel colorant (42.91-2863.4 µg/L) and detected in red ginseng concentrates containing >1% caramel colorant.

Identification of volatile markers for the detection of adulterants in red ginseng (Panax ginseng) juice using headspace stir-bar sorptive extraction coupled with gas chromatography and mass spectrometry.

Red ginseng (Panax ginseng) products are frequently adulterated by manufacturers with cheaper medicinal plant products including deodeok (Codonopsis lanceolata) and doraji (Platycodon grandiflorum) to increase profits. To identify possible volatile markers for the adulteration of red ginseng juices with deodeok or doraji, a headspace stir-bar sorptive extraction method was developed. Gas chromatography with mass spectrometry and untargeted metabolomics analysis revealed that 1-hexanol, cis-3-hexen-1-ol, and trans-2-hexen-1-ol are abundantly present in deodeok and doraji but not red ginseng. The peak area ratios in gas chromatograms of these compounds in red ginseng juices mixed with deodeok or doraji indicate that these volatile chemicals can be used as markers to detect the adulteration of red ginseng juice.

Highly Sensitive Detection of 4-Methylimidazole Using a Terahertz Metamaterial.

In this study, we demonstrated a highly sensitive detection method of 4-methylimidazole (4-MeI), a carcinogenic material, by using a terahertz (THz) metamaterial at a THz region. The THz metamaterials were fabricated with a metal array, using an electric-field-coupled inductor-capacitor (ELC) resonator structure, and a finite-difference time-domain (FDTD) simulation showed good agreement with the experimental results. We measured the THz spectra of the metamaterials to detect the 4-MeI concentrations of 0, 1, 2, 5, 10, 15, and 20 mg/L. The resonance frequency of the metamaterial was shifted by, approximately, 8 GHz and transmittance at the resonance frequency increased to 2 × 10-3, as the concentration was increased, up to 20 mg/L. Our study provides new insight into the application of metamaterials in detecting carcinogens, using a THz technique.

CAR1 deletion by CRISPR/Cas9 reduces formation of ethyl carbamate from ethanol fermentation by Saccharomyces cerevisiae.

Enormous advances in genome editing technology have been achieved in recent decades. Among newly born genome editing technologies, CRISPR/Cas9 is considered revolutionary because it is easy to use and highly precise for editing genes in target organisms. CRISPR/Cas9 technology has also been applied for removing unfavorable target genes. In this study, we used CRISPR/Cas9 technology to reduce ethyl carbamate (EC), a potential carcinogen, which was formed during the ethanol fermentation process by yeast. Because the yeast CAR1 gene encoding arginase is the key gene to form ethyl carbamate, we inactivated the yeast CAR1 gene by the complete deletion of the gene or the introduction of a nonsense mutation in the CAR1 locus using CRISPR/Cas9 technology. The engineered yeast strain showed a 98 % decrease in specific activity of arginase while displaying a comparable ethanol fermentation performance. In addition, the CAR1-inactivated mutants showed reduced formation of EC and urea, as compared to the parental yeast strain. Importantly, CRISPR/Cas9 technology enabled generation of a CAR1-inactivated yeast strains without leaving remnants of heterologous genes from a vector, suggesting that the engineered yeast by CRISPR/Cas9 technology might sidestep GMO regulation.

The effects of different extraction methods on essential oils from orange and tangor: From the peel to the essential oil.

Citrus fruits are largely consumed due to their unique and pleasant aromas. Citrus hybrids have been developed to enhance their flavors and bioactivities. Citrus aroma depends on the composition of the volatile compounds in citrus essential oils (CEOs), which are mostly located in the peels. During the extraction of CEOs, a specific series of chemical reactions occurred depending on the extraction methods (CP, cold pressing; HD, hydrodistillation), leading to variations in the composition of volatile compounds. In this study, the orange and the tangor which is a hybrid between C. reticulata × C. sinensis were investigated to compare the changes in volatile compounds based on the extraction methods. Results showed that the CP-specific volatile compounds were sesquiterpenes, oxygenated monoterpenes, and fatty acid derivatives, while the HD-specific volatile compounds were terpinyl cation derivatives, limonene, and 4-vinylguaiacol. On the other hand, the contents of some volatile compounds ((E)-ocimene, α-terpinene, and α-terpinolene) were affected by the Citrus species rather than by the extraction methods. In particular, during HD, terpinene-4-ol and 4-vinylguiacol, known as off-flavor compounds in citrus juice, were formed more abundantly in the orange than in the tangor. In conclusion, these results provide comprehensive data on essential oils, especially those derived from oranges and tangors, for selecting the appropriate extraction method for obtaining a higher yield and quality of citrus flavor.

Variation of volatile compounds and sensory profile for Protaetia brevitarsis larvae fermented with lactic acid bacteria and yeast.

This study aimed to investigate the correlation between the composition of volatile compounds, consumer acceptance, and drivers of (dis)liking of Protaetia brevitarsis larvae fermented using lactic acid bacteria and yeast. Volatile compounds were analyzed using HS-SPME-Arrow-GC-MS, and a sensory evaluation was conducted with 72 consumers. A total of 113 volatile compounds were detected, and principal component analysis indicated that the samples could be divided into three groups. The calculated relative odor activity values (ROAV) revealed the presence of 27 compounds (ROAV >1). Volatile compounds with high ROAV were predominantly found during yeast fermentation. The sensory evaluation results indicated a strong correlation between low levels of off-odor intensity and high odor liking, emphasizing that odor profile had a more direct association with consumer acceptance than odor intensity. These findings suggest that yeast fermentation using volatile compounds, which positively influences consumer acceptance, is appropriate for Protaetia brevitarsis larvae.

Stabilization of time domain acoustic boundary element method for the exterior problem avoiding the nonuniqueness.

The time domain boundary element method (TBEM) to calculate the exterior sound field using the Kirchhoff integral has difficulties in non-uniqueness and exponential divergence. In this work, a method to stabilize TBEM calculation for the exterior problem is suggested. The time domain CHIEF (Combined Helmholtz Integral Equation Formulation) method is newly formulated to suppress low order fictitious internal modes. This method constrains the surface Kirchhoff integral by forcing the pressures at the additional interior points to be zero when the shortest retarded time between boundary nodes and an interior point elapses. However, even after using the CHIEF method, the TBEM calculation suffers the exponential divergence due to the remaining unstable high order fictitious modes at frequencies higher than the frequency limit of the boundary element model. For complete stabilization, such troublesome modes are selectively adjusted by projecting the time response onto the eigenspace. In a test example for a transiently pulsating sphere, the final average error norm of the stabilized response compared to the analytic solution is 2.5%.

Headspace stir-bar sorptive extraction combined with gas chromatography-mass spectrometry for trace analysis of volatile organic compounds in Schisandra chinensis Baillon (omija).

Analyzing volatile organic compounds (VOCs) in food is crucial but challenging. Schisandra chinensis Baillon (omija) is an herbal plant with various functional health activities. Previous VOC analyses focused on S. chinensis fruit but not its leaves. Therefore, VOCs in S. chinensis fruit and leaves were analyzed using headspace stir-bar sorptive extraction (HS-SBSE)-GC-MS, and optimal conditions were established. Various factors, such as the sample preparation method, twister stir-bar type, sample amount, extraction temperature, and extraction time, expected to affect extraction were carefully optimized. Under the optimal conditions, 35 and 40 VOCs were identified in S. chinensis fruit and leaves, respectively. This HS-SBSE method is capable of rapid analysis and a low contamination rate without requiring organic solvents. These findings provide practical guidelines for HS-SBSE applications in various food matrices by providing analytical methods for VOC detection.

Determination of vitamin K encapsulated into lipid nanocarriers by dispersive liquid-liquid microextraction combined with liquid chromatography-atmospheric pressure chemical ionization-tandem mass spectrometry.

Dispersive liquid-liquid microextraction was used in conjunction with liquid chromatography-atmospheric pressure chemical ionization-tandem mass spectrometry to quantitate vitamins K1 and K2 in vitamin-fortified emulsions, and vital microextraction parameters were optimized using response surface methodology coupled with Box-Behnken design. Under optimal microextraction conditions, highly linear (R 2 > .999) calibration curves were obtained for both vitamins in a broad concentration range (1-1000 µg/L), and vitamin recoveries exceeded 90%. The detection and quantitation limits equaled 1.89 and 5.72 µg/L for vitamin K1, respectively, and 5.00 and 15.15 µg/L for vitamin K2, respectively. When applied to vitamin-K-loaded nanoemulsions and solid lipid nanoparticles, the developed method achieved excellent results, outperforming the currently employed Korean Food Code method, and therefore holding great promise for the quantitation of vitamin K in vitamin-fortified food products.

Stabilization of time domain acoustic boundary element method for the interior problem with impedance boundary conditions.

The time domain boundary element method (BEM) is associated with numerical instability that typically stems from the time marching scheme. In this work, a formulation of time domain BEM is derived to deal with all types of boundary conditions adopting a multi-input, multi-output, infinite impulse response structure. The fitted frequency domain impedance data are converted into a time domain expression as a form of an infinite impulse response filter, which can also invoke a modeling error. In the calculation, the response at each time step is projected onto the wave vector space of natural radiation modes, which can be obtained from the eigensolutions of the single iterative matrix. To stabilize the computation, unstable oscillatory modes are nullified, and the same decay rate is used for two nonoscillatory modes. As a test example, a transient sound field within a partially lined, parallelepiped box is used, within which a point source is excited by an octave band impulse. In comparison with the results of the inverse Fourier transform of a frequency domain BEM, the average of relative difference norm in the stabilized time response is found to be 4.4%.

Application of Edible Insects as Novel Protein Sources and Strategies for Improving Their Processing.

Insects have long been consumed by humans as a supplemental protein source, and interest in entomophagy has rapidly increased in recent years as a potential sustainable resource in the face of environmental challenges and global food shortages. However, food neophobia inhibits the widespread consumption of edible insects, despite their high nutritional and functional value. The own characteristics of edible insect protein such as foaming properties, emulsifying properties, gelling properties and essential amino acid ratio can be improved by drying, defatting, and extraction. Although nutritional value of some protein-enriched bread, pasta, and meat products, especially essential amino acid components was increased, replacement of conventional food with edible insects as a novel food source has been hindered owing to the poor cross-linking properties of edible insect protein. This deterioration in physicochemical properties may further limit the applicability of edible insects as food. Therefore, strategies must be developed to improve the quality of edible insect enriched food with physical, chemical, and biological methods. It was presented that an overview of the recent advancements in these approaches and highlight the challenges and prospects for this field. Applying these strategies to develop insect food in a more familiar form can help to make insect-enriched foods more appealing to consumers, facilitating their widespread consumption as a sustainable and nutritious protein source.

How to develop strategies to use insects as animal feed: digestibility, functionality, safety, and regulation.

Various insects have emerged as novel feed resources due to their economical, eco-friendly, and nutritive characteristics. Fish, poultry, and pigs are livestock that can feed on insects. The digestibility of insect-containing meals were presented by the species, life stage, nutritional component, and processing methods. Several studies have shown a reduced apparent digestibility coefficient (ADC) when insects were supplied as a replacement for commercial meals related to chitin. Although the expression of chitinase mRNA was present in several livestock, indigestible components in insects, such as chitin or fiber, could be a reason for the reduced ADC. However, various components can positively affect livestock health. Although the bio-functional properties of these components have been verified in vitro, they show positive health-promoting effects owing to their functional expression when directly applied to animal diets. Changes in the intestinal microbiota of animals, enhancement of immunity, and enhancement of antibacterial activity were confirmed as positive effects that can be obtained through insect diets. However, there are some issues with the safety of insects as feed. To increase the utility of insects as feed, microbial hazards, chemical hazards, and allergens should be regulated. The European Union, North America, East Asia, Australia, and Nigeria have established regulations regarding insect feed, which could enhance the utility of insects as novel feed resources for the future.

Physical and turbidimetric properties of cholecalciferol- and menaquinone-loaded lipid nanocarriers emulsified with polysorbate 80 and soy lecithin.

The present study aimed to investigate the physical and turbidimetric properties of cholecalciferol- and menaquinone-loaded lipid nanocarriers emulsified with different ratios of polysorbate 80 and soy lecithin. The lipid nanocarriers were subjected to three different heat treatments, LTLT (low temperature long time), HTST (high temperature short time), and autoclave treatments. Both cholecalciferol and menaquinone were successfully encapsulated in lipid nanocarriers and there was little loss of them during preparation. The droplet size of lipid nanocarriers emulsified with only polysorbate 80 increased and its PDI became larger than 0.3 after autoclave treatment. Moreover, 30.9% and 49.4% of cholecalciferol and menaquinone, respectively, were lost. In turbidimetric analysis, the destabilization by creaming formation in the upper layer of the lipid nanocarriers emulsified with a high polysorbate ratio was observed. However, the use of combination of both emulsifiers inhibited destabilization by flocculation as well as retained the cholecalciferol and menaquinone after all heat treatments.

CRISPR/Cas9-mediated Inactivation of arginase in a yeast strain isolated from Nuruk and its impact on the whole genome.

Despite the advantages of CRISPR/Cas9 technology in the food industry, controversy over its off-target effects exists. We engineered an industrial Saccharomyces cerevisiae strain isolated from a Korean rice wine starter, Nuruk, using CRISPR/Cas9 to decrease ethyl carbamate (EC) formation. We disrupted the CAR1 gene encoding arginase, which plays a key role in EC formation. Subsequently, we compared the whole genome of the engineered strain to that of the wild type by analyzing heterozygous and homozygous mutations through variant calling. Homozygous mutations in the genome of the engineered strains were identified as the target mutations in CAR1 induced by CRISPR/Cas9, and no other off-target effects were observed. Our findings have critical implications for the use of CRISRP/Cas9 technology in yeasts in the food industry.

Chemical Composition, Antioxidant, and Anti-Inflammatory Activity of Essential Oil from Omija (Schisandra chinensis (Turcz.) Baill.) Produced by Supercritical Fluid Extraction Using CO2.

Schisandra chinensis (Turcz.) Baill., which is known as omija in South Korea, is mainly cultivated in East Asia. The present study aimed to investigate the chemical composition of essential oil from the omija (OMEO) fruit obtained by supercritical fluid extraction using CO2 and to confirm the antioxidant and anti-inflammatory activity of OMEO using HaCaT human keratinocyte and RAW 264.7 murine macrophages. As a result of the chemical composition analysis of OMEO using gas chromatography-mass spectrometry, a total of 41 compounds were identified. The detailed analysis results are sesquiterpenoids (16), monoterpenoids (14), ketones (4), alcohols (3), aldehydes (2), acids (1), and aromatic hydrocarbons (1). OMEO significantly reduced the increased ROS levels in HaCaT keratinocytes induced by UV-B irradiation (p < 0.05). It was confirmed that 5 compounds (α-pinene, camphene, ß-myrcene, 2-nonanone, and nerolidol) present in OMEO exhibited inhibitory activity on ROS production. Furthermore, OMEO showed excellent anti-inflammatory activity in RAW 264.7 macrophages induced by lipopolysaccharide. OMEO effectively inhibited NO production (p < 0.05) by suppressing the expression of the iNOS protein. Finally, OMEO was investigated for exhibition of anti-inflammatory activity by inhibiting the activation of NF-κB pathway. Taken together, OMEO could be used as a functional food ingredient with excellent antioxidant and anti-inflammatory activity.

Analysis of volatile compounds in rooibos tea (Aspalathus linearis) using different extraction methods and their relationship with human sensory perception.

This study aimed to comparatively analyze the volatile flavor of rooibos tea (Aspalathus linearis) obtained by two commonly used flavor extraction methods, simultaneous distillation-extraction (SDE) and steam distillation under reduced pressure (DRP). The tea obtained by the two extraction methods, were analyzed by gas chromatography-mass spectrometry to identify volatile aroma-related compounds. Descriptive sensory analysis of the extracted rooibos tea flavor was carried out by a trained panel (n = 7). Fifty volatile compounds were identified, including 26 and 25 aroma-active compounds by SDE (45.9 µg/g) and DRP (37.5 µg/g), respectively. SDE recovered larger quantities of alcohols, acids, and esters, whereas DRP was useful for analyzing thermally unstable volatile compounds, including various alcohols, aldehydes, and hydrocarbons. Descriptive sensory analysis revealed that ketones and phenolic compounds may be responsible for the sensory attributes woody and grassy green, whereas the aldehydes and acidic compounds may contribute to floral and fruity.

Analyzing Gluten Content in Various Food Products Using Different Types of ELISA Test Kits.

Gluten is an insoluble protein produced when glutelins and prolamins, which are found in grains such as wheat, barley, and oats, combine to form an elastic thin film. This dietary gluten can cause severe contraction of the intestinal mucous membrane in some people, preventing nutrient absorption. This condition, called celiac disease (CD), affects approximately 1% of the world's population. The only current treatment for patients with CD and similar diseases is lifelong avoidance of gluten. To analyze the gluten content in food, various enzyme-linked immunosorbent assay (ELISA) tests are currently used. In this study, the gluten content in various food products was analyzed using different kinds of ELISA test kits. For gluten-free food, three different ELISA test kits mostly yielded values below the limit of detection. However, gluten was detected at 24.0-40.2 g/kg in bread, 6.5-72.6 g/kg in noodles, and 23.0-86.9 g/kg in different powder food samples. A significant difference (p < 0.05) in gluten content was observed for these gluten-containing food products. Reproducibility issues suggest that it is necessary to use several ELISA kits for the accurate detection and quantification of gluten in various food products rather than using one ELISA kit.