Proteomics analysis of enzyme systems and pathway changes during the moromi fermentation of soy sauce mash.

BACKGROUND: During the brewing of soy sauce, the conversion of multiple substances is driven by various microorganisms and their secreted enzyme systems. Soy sauce mash is an important source of enzyme systems during moromi fermentation, but the changes of enzyme systems in soy sauce mash during moromi fermentation are poorly understood. In order to explore the predominant enzyme systems existing during moromi fermentation and to explain the characteristics of the enzyme system changes, an enzymatic activities assay and 4D-label-free proteomics analysis were conducted on soy sauce mash at different stages of fermentation. RESULTS: The activities of hydrolytic enzymes in soy sauce mash decreased continuously throughout the fermentation process, while most of the characteristic physicochemical substances in soy sauce mash supernatant had already accumulated at the early stage of fermentation. Four hydrolytic enzymes were found to be positively correlated with important physicochemical indexes by principal component analysis and Pearson correlation analysis. The proteomics analysis revealed three highly upregulated enzymes and two enzymes that were present in important metabolic pathways throughout the fermentation process. Furthermore, it was found that Aspergillus oryzae was able to accumulate various nutrients in the soy sauce mash by downregulating most of its metabolic pathways. CONCLUSION: Enzymes present with excellent properties during the moromi fermentation period could be obtained from these results. Meanwhile, the characterization of the metabolic pathways of microorganisms during the moromi fermentation period was revealed. The results provide a basis for more scientific and purposeful improvement of moromi fermentation in the future. © 2024 Society of Chemical Industry.

Evaluation of the differences between low-salt solid-state fermented soy sauce and high-salt diluted-state fermented soy sauce in China: from taste-active compounds and aroma-active compounds to sensory characteristics.

BACKGROUND: The present study aimed to determine the components related to sensory properties in soy sauce and to characterize the differences between low-salt solid-state fermented soy sauce (LSFSS) and high-salt diluted-state fermented soy sauce (HDFSS). The taste and aroma active components of 18 commercially available soy sauces (eight types of LSFSS and 10 types of HDFSS) were characterized. The relationship between these compounds, soy sauce samples, and sensory properties was modeled by partial least squares regression. RESULTS: The analysis showed that the 11 taste-active components, including glutamic acid, glycine, alanine, threonine, malic acid, citric acid, tartaric acid, acetic acid, lactic acid, reducing sugar and salt, contributed greatly to the taste of soy sauce. In addition, umami, saltiness and sweetness are the characteristic tastes of HDFSS, whereas sourness and bitterness were the characteristic tastes of LSFSS. At the same time, seven aroma-active compounds, namely 4-ethyl-2-methoxyphenol, ethanol, 3-methyl-1-butanol, ethyl acetate, 2-phenethyl alcohol, 3-methyl thiopropanol and 2-ethyl-4-hydroxy-5-methylfuran-3-one, played a decisive role in the flavor of soy sauce. In addition, HDFSS presented the aroma attributes of smoky, alcoholic, floral, fruity and caramel-like, whereas LSFSS mainly presented sour and malty aroma attributes. CONCLUSION: The present study reveals new insight into the relationship between the chemical composition and sensory characteristics of soy sauce, which is of great significance for developing an objective measurement system and providing a theoretical basis to improve the sensory quality of soy sauce. © 2023 Society of Chemical Industry.

Identification of an operon and its regulator required for autoaggregation in Tetragenococcus halophilus.

IMPORTANCE: Tetragenococcus halophilus is a halophilic lactic acid bacterium generally used as a starter culture in fermenting soy and fish sauces. Aggregating strains can be useful in fermenting and obtaining clear soy sauce because cell clumps are trapped by the filter cake when the soy sauce mash is pressed. However, the genetic mechanisms of aggregation in T. halophilus are unknown. In this study, we identified genes encoding aggregation factor and its regulator. These findings may provide a foundation for developing improved T. halophilus starter cultures for soy sauce fermentation, leading to more efficient and consistent clear soy sauce production.

Ratio of Histamine-Producing/Non-Histamine-Producing Subgroups of Tetragenococcus halophilus Determines the Histamine Accumulation during Spontaneous Fermentation of Soy Sauce.

Strain specificity (within-species variation) of microorganisms occurs widely in nature. It might affect microbiome construction and function in a complex microbial environment. Tetragenococcus halophilus, a halophilic bacterium that generally is used in high salt food fermentation, consists of two histamine-producing and non-histamine-producing subgroups. It is unclear whether and how the strain specificity of histamine-producing capacity influences the microbial community function during food fermentation. Here, based on systematic bioinformatic analysis, histamine production dynamic analysis, clone library construction analysis, and cultivation-based identification, we identified that T. halophilus is the focal histamine-producing microorganism during soy sauce fermentation. Furthermore, we discovered that a larger number and ratio of histamine-producing subgroups of T. halophilus significantly contributed more histamine production. We were able to artificially decrease the ratio of histamine-producing to non-histamine-producing subgroups of T. halophilus in complex soy sauce microbiota and realized the reduction of histamine by 34%. This study emphasizes the significance of strain specificity in regulating microbiome function. This study investigated how strain specificity influenced microbial community function and developed an efficient technique for histamine control. IMPORTANCE Inhibiting the production of microbiological hazards under the assumption of stable and high-quality fermentation is a critical and time-consuming task for the food fermentation industry. For spontaneously fermented food, it can be realized theoretically by finding and controlling the focal hazard-producing microorganism in complex microbiota. This work used histamine control in soy sauce as a model and developed a system-level approach to identify and regulate the focal hazard-producing microorganism. We discovered that the strain specificity of focal hazard-producing microorganisms had an important impact on hazard accumulation. Microorganisms frequently exhibit strain specificity. Strain specificity is receiving increasing interest since it determines not only microbial robustness but also microbial community assembly and microbiome function. This study creatively explored how the strain specificity of microorganisms influenced microbiome function. In addition, we believe that this work provides an excellent model for microbiological hazard control which can promote future work in other systems.

Examining the impact of Tetragenococcus halophilus, Zygosaccharomyces rouxii, and Starmerella etchellsii on the quality of soy sauce: a comprehensive review of microbial population dynamics in fermentation.

Soy sauce is a popular fermented seasoning due to its distinct flavor and rich umami taste. Its traditional production involves two stages: solid-state fermentation and moromi (brine fermentation). During moromi, the dominant microbial population in the soy sauce mash changes, which is called microbial succession and is essential for the formation of soy sauce flavor compounds. Research has identified the sequence of succession, starting with Tetragenococcus halophilus, then Zygosaccharomyces rouxii, and lastly, Starmerella etchellsii. Factors such as the environment, microbial diversity, and interspecies relationships drive this process. Salt and ethanol tolerance influence microbial survival, while nutrients in the soy sauce mash support the cells in resisting external stress. Different microbial strains have varying abilities to survive and respond to external factors during fermentation, which impacts soy sauce quality. In this review, we would examine the factors behind the succession of common microbial populations in the soy sauce mash and explore how microbial succession affects soy sauce quality. The insights gained can help better manage the dynamic changes in microbes during fermentation, leading to improved production efficiency.

Six categories of amino acid derivatives with potential taste contributions: a review of studies on soy sauce.

During the fermentation of soy sauce, the metabolism of microorganisms and the Maillard reaction produce a wide variety of metabolites that contribute to the unique and rich flavor characteristics of soy sauce, such as amino acids, organic acids and peptides. Amino acid derivatives, a relatively new taste compounds, formed by the reaction of enzymes or non-enzymes from sugars, amino acids, and organic acids released through metabolism by microorganisms during soy sauce fermentation, have begun to gain more and more attention in recent years. This review focused on our existing knowledge of the sources, taste characteristics and synthesis methods of the 6 categories of amino acid derivatives, including Amadori compounds, γ-glutamyl peptides, pyroglutamyl amino acids, N-lactoyl amino acids, N-acetyl amino acids and N-succinyl amino acids. Sixty-four amino acid derivatives were detected in soy sauce, of which 47 were confirmed to have potential contribution to the taste of soy sauce, especially umami and kokumi, and some of them also have the effect of reducing bitterness. Furthermore, some amino acid derivatives, like γ-glutamyl peptides and N-lactoyl amino acids, were found to be synthesized enzymatically in vitro, which laid the foundation for further study on their formation pathways in the future.

Fluorescent Carbon Dots Derived From Soy Sauce for Picric Acid Detection and Cell Imaging.

Picric acid (PA) is a powerful nitro-aromatic explosive that harms the environment and human health. Developing non-toxic and low-cost sensors for the rapid detection of PA is essential. An environment-friendly fluorescent probe for PA detection is designed based on carbon dots (CDs) directly separated from edible soy sauce by silica gel column chromatography. Neither organic reagents nor heating process was needed to prepare CDs. The obtained CDs exhibit bright blue fluorescence, good water solubility, and photostability. The fluorescent probe for PA was developed according to the CD's fluorescence can be significantly quenched via the inner filter effect between CDs and PA. The linear range was 0.2-24 µM with a limit of detection of 70 nM. This proposed method was successfully employed to detect PA in the real water samples with satisfactory recoveries between 98.0-104.0%. Moreover, the CDs were suitable for fluorescence imaging of HeLa cells owing to their low toxicity and good biocompatibility.

CRISPR/Cas9 genome editing for comparative genetic analysis related to soy sauce brewing in Aspergillus sojae industrial strains.

Aspergillus sojae has traditionally been used in soy sauce brewing. Genetic modification techniques have been established in A. sojae, but it is difficult to apply them to various industrial strains. Although we have previously developed a CRISPR/Cpf1 system for genetic modification of A. sojae, another genome editing system was required for versatile modification. In addition, repetitive genetic modification using the CRISPR system has not been established in A. sojae. In this study, we demonstrated mutagenesis, gene deletion/integration, and large deletion of a chromosomal region in A. sojae using the CRISPR/Cas9 system. We also successfully performed repetitive genetic modification using a method that involved forced recycling of genome-editing plasmids. Moreover, we demonstrated that the effects of genetic modification related to soy sauce brewing differed among A. sojae industrial strains. These results showed that our technique of using the CRISPR/Cas9 system is a powerful tool for genetic modification in A. sojae.

Correlation between bacterial diversity and flavor substances in Longgang soy sauce.

Longgang soy sauce is one of the traditional fermented condiments in China, but its bacterial community succession and its unique flavor development during the fermentation process are not well-investigated. This study evaluated the bacterial diversity, flavor changes, and their correlation during the fermentation of Longgang soy sauce. The results showed that Weissella was the dominant bacterial genus in the fermentation stage of sauce fermented grains. In the first 31 days of the moromi fermentation stage, a variety of bacterial genera such as Weissella, Halomonas, Bacteroides, Pseudomonas, and Tetragenococcus were the dominant bacteria. Our results showed that these bacteria have a significantly positive correlation with phenylethyl alcohol, ethyl acetate, and 3-methyl-1-butanol. As the fermentation progressed, a flora structure with Halomonas as the main bacterial genus was formed. This genus exhibited a significantly positive and positive correlation with 1-octanol, ethyl palmitate, heptanol, and 2-nonanol, which are the unique flavor components of Longgang soy sauce.

ß-Carboline Alkaloids in Soy Sauce and Inhibition of Monoamine Oxidase (MAO).

Monoamine oxidase (MAO) oxidizes neurotransmitters and xenobiotic amines, including vasopressor and neurotoxic amines such as the MPTP neurotoxin. Its inhibitors are useful as antidepressants and neuroprotectants. This work shows that diluted soy sauce (1/3) and soy sauce extracts inhibited human MAO-A and -B isozymes in vitro, which were measured with a chromatographic assay to avoid interferences, and it suggests the presence of MAO inhibitors. Chromatographic and spectrometric studies showed the occurrence of the ß-carboline alkaloids harman and norharman in soy sauce extracts inhibiting MAO-A. Harman was isolated from soy sauce, and it was a potent and competitive inhibitor of MAO-A (0.4 µM, 44 % inhibition). The concentrations of harman and norharman were determined in commercial soy sauces, reaching 243 and 52 µg/L, respectively. Subsequently, the alkaloids 1,2,3,4-tetrahydro-ß-carboline-3-carboxylic acid (THCA) and 1-methyl-1,2,3,4-tetrahydro-ß-carboline-3-carboxylic acid (MTCA) were identified and analyzed in soy sauces reaching concentrations of 69 and 448 mg/L, respectively. The results show that MTCA was a precursor of harman under oxidative and heating conditions, and soy sauces increased the amount of harman under those conditions. This work shows that soy sauce contains bioactive ß-carbolines and constitutes a dietary source of MAO-A and -B inhibitors.

Flavor differences of soybean and defatted soybean fermented soy sauce and its correlation with the enzyme profiles of the kojis.

BACKGROUND: Soybeans and defatted soybeans, commonly used as protein ingredients, have different flavors of their fermented soy sauce. Clarifying the differences between the two soy sauces, as well as the formation mechanism, is an important prerequisite for improving the flavor of defatted soybean soy sauce. To this goal, the aroma characteristics of two soy sauces and their volatile profiles were compared by sensory evaluation and gas chromatography-mass spectrometry, and eight enzyme activities and volatile profiles of matured koji were determined. RESULTS: Sensory results showed that the acids, fruity and cooked potato-like attributes were higher in whole soybean fermented soy sauce, whereas defatted soybean soy sauce exhibited higher smoky and malty attributes, closely related to the contents of aroma-active compounds in soy sauce, such as isobutyl acetate, 2/3-methylbutanal, acetic acid and 2/3-methylbutanoic acid. The content of most volatiles in the matured kojis showed a consistent trend with that of soy sauce: alcohols, acids, furan(one)s and ketones. Interestingly, acid protease and cellulase activities were 3.3 and 1.6 times higher in the whole soybean koji than in defatted soybean koji, respectively, whereas neutral protease, aminopeptidase, glucoamylase and ß-glucosidase were approximately 2.0 times higher in defatted soybean koji. CONCLUSION: In summary, the flavor differences between soybean and defatted soybean fermented soy sauce were not only caused by the differences in the content of flavor precursors in the materials, but also closely related to the differences in the enzymatic profiles accumulated during the koji-making process. © 2022 Society of Chemical Industry.

Changes in quality components and antioxidant activity of peony seed soy sauce during low-salt solid-state fermentation.

BACKGROUND: In this study, the fermentation conditions of peony seed soy sauce (PSSS) koji were optimized by response surface method, and the quality components and antioxidant activity of PSSS were investigated at different low-salt solid-state fermentation stages. RESULTS: Results of response surface method showed that the optimal fermentation conditions were 460.6 g kg-1 water content, 48.6 h culture time, 31.5 °C culture temperature and ratio 2.1:1 (w/w) of peony seed meal:wheat bran, with the highest neutral protease activity (2193.78 U g-1 ) of PSSS koji. PSSS had the highest amino acid nitrogen (7.69 g L-1 ), salt-free soluble solids (185.26 g L-1 ), total free amino acids (49.03 g L-1 ), essential free amino acids (19.58 g L-1 ) and umami free amino acids (16.64 g L-1 ) at 20 days of fermentation. The highest total phenolics were 5.414 g gallic acid equivalent L-1 and total flavonoids 0.617 g rutin equivalent L-1 , as well as the highest DPPH radical scavenging activity (86.19%) and reducing power (0.8802, A700 ) of PSSS fermented at 30 days. Sensory evaluation showed that fermentation of 20 days and 25 days could produce a better taste and aroma of PSSS than 15 days and 30 days. CONCLUSION: PSSS had the highest quality components in the middle of fermentation (20 days) and the highest antioxidant activity in the late fermentation period (30 days). These results demonstrated that peony seed meal could be used to produce high-quality soy sauce with high antioxidant activity. © 2023 Society of Chemical Industry.

[Development for the Simultaneous Analytical Method of Furan and Alkyl Furans in Processed Foods].

A simultaneous analytical method was developed for the determination of alkyl furans (Furan, 2-methylfuran, 3-methylfuran and 2,5-dimethylfuran) in processed foods by headspace-GC-MS. Single-laboratory validation data of furan, 2-methylfuran, 3-methylfuran and 2,5-dimethylfuran showed good precision and accuracy. The mean recoveries ranged from 92 to 116%, the intermediate precision (RSDi) ranged from 0.9 to 12.9%. The level of LOQ ranged from 0.5 to 1.2 µg/kg (coffee), from 3.5 to 4.1 µg/kg (soy sauce), from 0.4 to 1.3 µg/kg (other foods: clear apple juice, infant formula and baby food), respectively. This method has the sensitivity to detect low levels of furan and alkyl furans contaminated in various foods and is thus applicable to surveillance for risk management in food safety.

[Cost-benefit analysis on the intervention of application iron-fortified soy sauce for anemia in Deqing 15-54 years women, Zhejiang].

OBJECTIVE: To make a cost-benefit analysis on anemia intervention with iron-fortified soy sauce in 15-54 years old women. METHODS: The study was conducted in Deqing county, Zhejiang province in 2012-2013. A total 585 women as sampling size were estimated with statistical model and randomly selected by probability proportionate to size sampling. Hemoglobin were measured before intervention and after 15 months. The cost of the intervention project were collected with manpower, communication and other invest. The benefit was estimated with profiling model. RESULTS: After the intervention, the anemia prevalence of sampled women decreased from 31.1% to 21.9%(P<0.01). The major cost of the project was 156 400 RMB, and total benefits result ing from projects were 1 448 485 RMB. The cost-benefit ratio of the project is 1∶9.49. If investing one yuan can produce economic benefits of nearly 9.49 yuan, therefore, the intervention projectis worth to be scaling up. Sensitivity analysis showed the result of this study was stable. CONCLUSION: The intervention can significantly reduce the prevalence of anemia in women, and reduce the economic burden of the diseases. .

Multi-omics analyses of the mechanism for the formation of soy sauce-like and soybean flavor in Bacillus subtilis BJ3-2.

Although soy sauce-like flavor and soybean flavor are two key contributors to the flavor of fermented foods, the key compounds of soy sauce-like flavor and soybean flavor and production mechanisms are still poorly understood and need further investigation. In the present study, we found that the Bacillus subtilis (B. subtilis) BJ3-2 strain has various metabolic properties at different temperatures, and the strain cultured at 37℃ increased the soybean flavor (a special flavor of ammonia-containing smelly distinct from natto) compared with culturing at 45℃ and 53℃. Interestingly, the strain cultured at 45℃ and 53℃ had a higher soy sauce-like flavor than that in 37℃. Moreover, a comparative transcriptome analysis of the strain cultured at 37℃, 45℃, and 53℃ showed transcriptional changes related to secondary metabolites and ABC transporters, which is critical for the amino acid transport and metabolism in B. subtilis. Meanwhile, proteomics and metabolomics profiling showed a marked change in amino acids transport and metabolism. In addition, the metabolic analysis revealed a significant metabolic difference (including sulfur metabolism, glutathione metabolism, nicotinate and nicotinamide metabolism, cysteine and methionine metabolism, and pyrimidine metabolism) in the strain cultured at 45℃ and 53℃ compared to 37℃. To sum, this study used the multi-omics profiling tool to investigate the fermentative strains B. subtilis BJ3-2, thus providing a deeper insight into the mechanism of the formation of soy sauce-like flavor and soybean flavor compounds.

Genomic features, aroma profiles, and probiotic potential of the Debaryomyces hansenii species complex strains isolated from Korean soybean fermented food.

Fermented soybean products are gaining attention in the food industry owing to their nutritive value and health benefits. In this study, we performed genomic analysis and physiological characterization of two Debaryomyces spp. yeast isolates obtained from a Korean traditional fermented soy sauce "ganjang". Both Debaryomyces hansenii ganjang isolates KD2 and C11 showed halotolerance to concentrations of up to 15% NaCl and improved growth in the presence of salt. Ploidy and whole-genome sequencing analyses indicated that the KD2 genome is haploid, whereas the C11 genome is heterozygous diploid with two distinctive subgenomes. Interestingly, phylogenetic analysis using intron sequences indicated that the C11 strain was generated via hybridization between D. hansenii and D. tyrocola ancestor strains. The D. hansenii KD2 and D. hansenii-hybrid C11 produced various volatile flavor compounds associated with butter, caramel, cheese, and fruits, and showed high bioconversion activity from ferulic acid to 4-vinylguaiacol, a characteristic flavor compound of soybean products. Both KD2 and C11 exhibited viability in the presence of bile salts and at low pH and showed immunomodulatory activity to induce high levels of the anti-inflammatory cytokine IL-10. The safety of the yeast isolates was confirmed by analyzing virulence and acute oral toxicity. Together, the D. hansenii ganjang isolates possess physiological properties beneficial for improving the flavor and nutritional value of fermented products.

Comparative study on the novel umami-active peptides of the whole soybeans and the defatted soybeans fermented soy sauce.

BACKGROUND: Two kinds of soy sauce produced via fermentation of the whole soybeans and the defatted soybeans (soy sauce termed 'SSS' and 'SSD', respectively) were subjected to the treatment using aqueous ethanol solutions with high concentrations. Then tasty peptides were separated from SSS and SSD by sensory guided fractionation, using macroporous resin and reverse-phase high-performance liquid chromatography (RP-HPLC), and identified by ultra-performance liquid chromatography quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS). RESULTS: The result showed that umami-active fractions and ≤ 3 kDa peptides were mainly concentrated in the supernatants resulted from the treatment with 60% ethanol and the precipitates resulted from the treatment with 80% ethanol. The contents of ammonia nitrogen, non-salt solids, bitter amino acids, amino acids in peptides and the proportion of ≤ 3 kDa peptides in SSS were higher than those in SSD. Sixteen and fourteen tasty peptides were separated from SSS and SSD, among them five dipeptides (γ-Glu-Glu, Glu-Glu, γ-Glu-Cys, γ-Glu-Leu, Glu-Leu and Ile-Glu) with a glutamic acid residue were identified both in SSS and SSD, which have been reported as umami/kokumi-active peptides in soy sauce. Several peptides identified from SSS (Thr-Gly-Cys, Gly-Leu-Glu, Val-Glu-Ala-Leu and Gly-Gly-Gly-Glu) and SSD (Asp-Arg, Asp-Ala-Glu, Glu-Val-Cys and Gly-Gly-Gly-Glu) are tasty and/or umami-active peptides but have not been reported as tasty peptides in the past. CONCLUSION: Most of the peptides separated from SSD and SSS could impart an umami-enhancing effect on soy sauce, and the marginally more kokumi and bitterness hydrophobic peptides were found in SSS than SSD. © 2020 Society of Chemical Industry.

Combination of mutagenesis and adaptive evolution to engineer salt-tolerant and aroma-producing yeast for soy sauce fermentation.

BACKGROUND: The moromi fermentation of high-salt liquid-state fermentation (HLF) soy sauce is usually performed in high-brine solution (17-20%, w/w), which decreases the metabolic activity of aroma-producing yeast. To enhance the soy sauce flavors, increasing the salt tolerance of aroma-producing yeasts is very important for HLF soy sauce fermentation. RESULTS: In the present study, atmospheric and room-temperature plasma (ARTP) was first used to mutate the aroma-producing yeast Wickerhamomyces anomalus, and the salt tolerant strains were obtained by selection of synthetic medium with a sodium chloride concentration of 18% (w/w). Furthermore, adaptive laboratory evolution (ALE) was used to improve the salt tolerance of the mutant strains. The results obtained indicated that the combination use of ARTP and ALE markedly increased the NaCl tolerance of the yeast by increasing the cellular accumulation of K+ and removal of cytosolic Na+ , in addition to promoting the production of glycerin and strengthening the integrity of the cell membrane and cell wall. In soy sauce fermentation, the engineered strains improved the physicochemical parameters of HLF soy sauce compared to those produced by the wild-type strain, and the engineered strains also increased the alcohol, acid and aldehyde production, and enriched the types of esters in the soy sauce. CONCLUSION: The results of the present study indicated that the combination of ARTP mutagenesis and ALE significantly improved the salt tolerance of the aroma-producing yeast, and also enhanced the production of volatiles of HLF soy sauce. © 2021 Society of Chemical Industry.

[Effect of Soy Sauce, Fish Sauce for Nitrite Determination in Meat Products].

The diazotization method is used for nitrite determination in meat products. In this method, it is known that the presence of reducing substances such as ascorbic acid and cysteine in the sample interferes with nitrite determination. We speculated that soy sauce, fish sauce and mirin, which are used in meat products, might be the reducing substances, so investigated meat products which contain soy sauce and other ingredients. The results showed that quantitative disturbance was observed in soy sauce and fish sauce, and the recovery in spiked samples of these products decreased in proportion to the increase in soy sauce and fish sauce content. However the antioxidant of soy sauce and fish sauce depending on manufacturing methods, so quantitative disturbance dose not be observed each time, so nitrite determination in meat products, and to conduct simultaneous determination using different sample amounts to confirm whether there is any difference in the quantitative values.

Comparison of trapping efficiency of dicarbonyl trapping agents and reducing agents on reduction of furanoic compounds in commercially available soy sauce varieties.

This study has conducted to determine the concentration levels of furan, furfural and 2-methylfuran in the six commercially available soy sauce varieties by an optimized Gas Chromatography Tripple Quadruple Mass Spectrometry. The extraction of analytes was performed by solid phase microextraction using 50/30 µm CAR/DVB/PDMS fibre in 25 min with 20% NaCl concentration under 35 °C and separation was performed on HP5-MS column. Different concentration levels of furan, furfural and 2-methylfuran were determined which differed significantly at p < 0.05. A total of four metal ions (Iron sulphate, magnesium sulphate, calcium sulphate and sodium sulfite), ascorbic acid and natural antioxidants (epicatechin, epigalactocatechin and kempferol) were added in the soy sauce samples by simulating sterilization conditions. A higher reduction level was given by calcium sulphate with reduction upto 90.68%, 89.07% and 65.42% for furan, furfural and 2-methylfuran, respectively, in comparison with other metal ions. Since iron sulphate and ascorbic acid have triggered formation of furanoic compounds upto high levels instead of reduction. Moreover, from natural antioxidants, epicatechin and kemferol have provided more reduction levels around 89.66%, 90.14% and 78.75% for furan, furfural and 2-methylfuran, respectively by sterilization with catichen and 88.80%, 90.36% and 84.29% for furan, furfural and 2-methylfuran, respectively by sterilization with kempferol than epigalactocatechin. Moreover, this method was also validated in terms of sensitivity, recovery, relative standard deviation and LOD and LOQ for all analytes.