Measurements of elemental iodine in soy sauces in Taiwan using a modified microplate method.

Background: Soy sauce is widely used in a variety of Asian dishes to enhance flavor. Soybean and most soybean products, including soy sauces, are listed as prohibited foods in a low iodine diet. However, the iodine content in soy sauces is largely unknown. The aim of this study was to determine the iodine content in domestic soy sauces in Taiwan. Methods: Twenty-five different kinds of soy sauces were diluted with distilled water and with a dilution factor of fifty or above. Iodine concentrations of the diluted samples were measured colourimetrically based on the Sandell-Kolthoff reaction by a modified microplate method. All the measurements were repeated twelve times on three different days for determination of mean and standard deviation (SD), and coefficients of variance (CV). Serial dilution and recovery tests were also performed for validation. The results were confirmed by an inductively coupled plasma mass spectrometry (ICP-MS) method. Results: Among the twenty-five surveyed soy sauces, most of them (n=22) were iodine-free (<16 ug/L, and thus un-detectable). The iodine concentrations (mean ± SD) of the three iodine-containing soy sauces were 2.7 ± 0.1, 5.1 ± 0.2, and 10.8 ± 0.6 mg/L, respectively. The inter-assay, intra-assay and total CVs were all <5.3% for the modified microplate method. The results obtained by ICP-MS were consistent with those of the modified microplate method. The recovery rates in the serial dilution test and recovery test ranged from 94.7% to 118.6%. Two of the three iodine-containing soy sauces were supplemented with kelp extract, while the other one without kelp extract had the highest amount of salt among the three iodine-containing soy sauces. Therefore, we postulate that iodized salt instead of kelp extract is the source of higher iodine content in that sauce. Conclusion: The results suggest that most soy sauces are iodine-free and may be allowed during low iodine diets.

Developmental exposure to phytoestrogens found in soy: New findings and clinical implications.

Exposure to naturally derived estrogen receptor activators, such as the phytoestrogen genistein, can occur at physiologically relevant concentrations in the human diet. Soy-based infant formulas are of particular concern because infants consuming these products have serum genistein levels almost 20 times greater than those seen in vegetarian adults. Comparable exposures in animal studies have adverse physiologic effects. The timing of exposure is particularly concerning because infants undergo a steroid hormone-sensitive period termed "minipuberty" during which estrogenic chemical exposure may alter normal reproductive tissue patterning and function. The delay between genistein exposure and reproductive outcomes poses a unique challenge to collecting epidemiological data. In 2010, the U.S. National Toxicology Program monograph on the safety of the use of soy formula stated that the use of soy-based infant formula posed minimal concern and emphasized a lack of data from human subjects. Since then, several new human and animal studies have advanced our epidemiological and mechanistic understanding of the risks and benefits of phytoestrogen exposure. Here we aim to identify clinically relevant findings regarding phytoestrogen exposure and female reproductive outcomes from the past 10 years, with a focus on the phytoestrogen genistein, and explore the implications of these findings for soy infant formula recommendations. Research presented in this review will inform clinical practice and dietary recommendations for infants based on evidence from both clinical epidemiology and basic research advances in endocrinology and developmental biology from mechanistic in vitro and animal studies.

Dietary Ceramide Prepared from Soy Sauce Lees Improves Skin Barrier Function in Hairless Mice.

Dietary sphingolipids such as glucosylceramide and sphingomyelin are known to improve the skin barrier function of damaged skin. In this study, we focused on free-ceramide prepared from soy sauce lees, which is a byproduct of soy sauce production. The effects of dietary soy sauce lees ceramide on the skin of normal mice were evaluated and compared with those of dietary maize glucosylceramide. We found that transepidermal water loss value was significantly suppressed by dietary supplementation with soy sauce lees ceramide as effectively as or more effectively than maize glucosylceramide. Although the content of total and each subclass of ceramide in the epidermis was not significantly altered by dietary sphingolipids, that of 12 types of ceramide molecules, which were not present in dietary sources, was significantly increased upon ingestion of maize glucosylceramide and showed a tendency to increase with soy sauce lees ceramide intake. In addition, the mRNA expression of ceramide synthase 4 and involucrin in the skin was downregulated by sphingolipids. This study, for the first time, demonstrated that dietary soy sauce lees ceramide enhances skin barrier function in normal hairless mice, although further studies are needed to clarify the molecular mechanism.

United States Pharmacopeia (USP) Safety Review of Gamma-Aminobutyric Acid (GABA).

Gamma-amino butyric acid (GABA) is marketed in the U.S. as a dietary supplement. USP conducted a comprehensive safety evaluation of GABA by assessing clinical studies, adverse event information, and toxicology data. Clinical studies investigated the effect of pure GABA as a dietary supplement or as a natural constituent of fermented milk or soy matrices. Data showed no serious adverse events associated with GABA at intakes up to 18 g/d for 4 days and in longer studies at intakes of 120 mg/d for 12 weeks. Some studies showed that GABA was associated with a transient and moderate drop in blood pressure (<10% change). No studies were available on effects of GABA during pregnancy and lactation, and no case reports or spontaneous adverse events associated with GABA were found. Chronic administration of GABA to rats and dogs at doses up to 1 g/kg/day showed no signs of toxicity. Because some studies showed that GABA was associated with decreases in blood pressure, it is conceivable that concurrent use of GABA with anti-hypertensive medications could increase risk of hypotension. Caution is advised for pregnant and lactating women since GABA can affect neurotransmitters and the endocrine system, i.e., increases in growth hormone and prolactin levels.

Potential Health Benefits Associated with Lunasin Concentration in Dietary Supplements and Lunasin-Enriched Soy Extract.

Lunasin has demonstrated antioxidative, anti-inflammatory, and chemopreventive properties. The objectives were to evaluate the concentration of lunasin in different lunasin-based commercial dietary supplements, to produce a lunasin-enriched soy extract (LESE) using a two-step pilot-plant-based ultrafiltration process, and to evaluate their biological potential in vitro. LESE was produced using 30 and 1 kDa membranes in a custom-made ultrafiltration skid. Lunasin was quantified in eight products and LESE. Lunasin concentrations of the lunasin-based products ranged from 9.2 ± 0.6 to 25.7 ± 1.1 mg lunasin/g protein. The LESE extract contained 58.2 mg lunasin/g protein, up to 6.3-fold higher lunasin enrichment than lunasin-based dietary supplements. Antioxidant capacity ranged from 121.5 mmol Trolox equivalents (TE)/g in Now® Kids to 354.4 mmol TE/g in LESE. Histone acetyltransferase (HAT) inhibition ranged from 5.3% on Soy Sentials® to 38.3% on synthetic lunasin. ORAC and lunasin concentrations were positively correlated, and HAT and lunasin concentrations were negatively correlated (p < 0.05). Melanoma B16-F10 and A375 cells treated with lunasin showed dose-dependent inhibitory potential (IC50 equivalent to 330 and 370 µM lunasin, respectively). Lunasin showed protein kinase B expression (57 ± 14%) compared to the control (100%) in B16-F10. Lunasin concentration found in commercial products and lunasin-enriched soy extract could exert benefits to consumers.

A metabolomics approach to evaluate post-fermentation enhancement of daidzein and genistein in a green okara extract.

BACKGROUND: Okara is a major agri-industrial by-product of the tofu and soymilk industries. Employing food-wastes as substrates for the green production of natural functional compounds is a recent trend that addresses the dual concepts of sustainable production and a zero-waste ecosystem. RESULTS: Extracts of unfermented okara and okara fermented with Rhizopus oligosporus were obtained using ethanol as extraction solvent, coupled with ultrasound sonication for enhanced extraction. Fermented extracts yielded significantly better results for total phenolic content (TPC) and total flavonoid content (TFC) than unfermented extracts. A qualitative liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QTOF-MS) analysis revealed a shift from glucoside forms to respective aglycone forms of the detected isoflavones, post-fermentation. Since the aglycone forms have been associated with numerous health benefits, a quantitative high-performance liquid chromatography (HPLC) analysis was performed. Fermented okara extracts had daidzein and genistein concentrations of 11.782 ± 0.325 µg mL-1 and 10.125 ± 1.028 µg mL-1 , as opposed to that of 6.7 ± 2.42 µg mL-1 and 4.55 ± 0.316 µg mL-1 in raw okara extracts, respectively. Lastly, the detected isoflavones were mapped to their metabolic pathways, to understand the biochemical reactions triggered during the fermentation process. CONCLUSION: Fermented okara may be implemented as a sustainable solution for production of natural bioactive isoflavonoids genistein and daidzein. © 2021 Society of Chemical Industry.

Effects of the extract from peanut meal fermented with Bacillus natto and Monascus on lipid metabolism and intestinal barrier function of hyperlipidemic mice.

BACKGROUND: Hyperlipidemia is one of the metabolic disorders that poses a great threat to human health. This study is aimed at investigating the potential hypolipidemic properties of extract from peanut meal fermented with Bacillus natto and Monascus in mice fed with a high-fat diet. Herein, 60 male C57BL/6J mice were randomly divided into six groups: four control groups, comprised of a normal group, a model (M) group, a positive control group (atorvastatin 10 mg kg-1 ), and a nonfermented peanut meal extract group (150 mg kg-1 ), and two experimental groups, comprised of a fermented peanut meal extract low-dose group (50 mg kg-1 ) and a fermented peanut meal extract high-dose group (FH, 150 mg kg-1 ). RESULTS: Body weight (P = 0.001) and levels of serum total cholesterol (P = 0.007), triacylglycerol (P = 0.040), low-density lipoprotein cholesterol (P < 0.001), and leptin (P < 0.001) were remarkably decreased in the FH group, whereas the serum high-density lipoprotein cholesterol levels were increased (P < 0.001) by 78.3% compared with the M group. Ileum tissue stained with hematoxylin and eosin showed that the ileal villus detachments in mice were improved, and the villus height was increased by supplementation with extract from fermented peanut meal. Moreover, the expressions of intestinal ZO-1 (P = 0.003) and occludin (P = 0.013) were elevated in the FH group, compared with the M group. CONCLUSION: Extract of peanut meal fermented by B. natto and Monascus can effectively improve hyperlipidemia caused by a high-fat diet in mice, via regulating leptin and blood lipid levels, and protect the intestinal mucosal barrier, which provides evidence for its anti-hyperlipidemia effects and is a research basis for potential industrial development. © 2020 Society of Chemical Industry.

Tartary buckwheat protein hydrolysates enhance the salt tolerance of the soy sauce fermentation yeast Zygosaccharomyces rouxii.

Supplementation of protein hydrolysate is an important strategy to improve the salt tolerance of soy sauce aroma-producing yeast. In the present study, Tartary buckwheat protein hydrolysates (BPHs) were prepared and separated by ultrafiltration into LM-1 (<1 kDa) and HM-2 (1-300 kDa) fractions. The supplementation of HM-2 fraction could significantly improve cell growth and fermentation of soy sauce aroma-producing yeast Zygosaccharomyces rouxii As2.180 under high salt (12%, w/w) conditions. However, the LM-1 fraction inhibited strain growth and fermentation. The addition of HM-2 promoted yeast cell accumulation of K+, removal of cytosolic Na+ and accumulation of glycerol. Furthermore, the HM-2 fraction improved the cell membrane integrity and mitochondrial membrane and decreased intracellular ROS accumulation of the strain. The above results indicated that the supplementation of BPHs with a molecular weight of 1-300 kDa is a potentially effective and feasible strategy for improving the salt tolerance of soy sauce aroma-producing yeast Z. rouxii.

Phytoestrogen intake and other dietary risk factors for low motile sperm count and poor sperm morphology.

BACKGROUND: Few potentially modifiable risk factors of male infertility have been identified, and while different diets and food groups have been associated with male infertility, evidence linking dietary factors including phytoestrogens and semen quality is limited and contradictory. OBJECTIVES: To study the associations between phytoestrogen intake and other dietary factors and semen quality. MATERIALS AND METHODS: A case-referent study was undertaken of the male partners, of couples attempting conception with unprotected intercourse for 12 months or more without success, recruited from 14 UK assisted reproduction clinics. A total of 1907 participants completed occupational, lifestyle and dietary questionnaires before semen quality (concentration, motility and morphology) were assessed. Food intake was estimated by a 65-item food frequency questionnaire (FFQ) covering the 12 months prior to recruitment. Analyses of dietary risk factors for low motile sperm concentration (MSC: <4.8 × 106 /mL) and poor sperm morphology (PM: <4% normal morphology) used unconditional logistic regression, accounting for clustering of subjects within the clinics, first without, and then with, adjustment for confounders associated with that outcome. RESULTS: High consumption of daidzein (≥13.74 µg/d), a phytoestrogen found in soy products, was a protective factor for MSC with an odds ratio (95%CI) of 0.58 (0.42-0.82) after adjustment for clustering and potential confounding. Dietary risk factors for PM after similar adjustment showed that drinking whole milk (OR 0.67, 95%CI 0.47-0.96) and eating red meat were protective with an OR 0.67 (0.46-0.99) for eating red meat >3 times/wk. DISCUSSION: In this case-referent study of men attending an infertility clinic for fertility diagnosis, we have identified that low MSC is inversely associated with daidzein intake. In contrast, daidzein intake was not associated with PM but eating red milk and drinking whole milk were protective. CONCLUSIONS: Dietary factors associated with semen quality were identified, suggesting that male fertility might be improved by dietary changes.

Isolation and identification of the antimicrobial substance included in tempeh using Rhizopus stolonifer NBRC 30816 for fermentation.

In this study, we focus on the antimicrobial properties of tempeh, a soybean fermented food, against oral bacteria. Tempeh showed antimicrobial activity against dental caries pathogenic bacterium Streptococcus mutans at a final concentration of 1 mg/mL. An antimicrobial substance contained in tempeh was present in the 100 kDa or greater fraction generated by ultrafiltration, but it was found not to be proteinaceous by native-PAGE, SDS-PAGE and protein degradation tests. Next, when the fraction was purified with an ODS column, the 80% and 100% methanol eluates showed antimicrobial activity against S. mutans. The 100% methanol eluate was further subjected to a 2nd column purification, and isolation of the target was confirmed by HPLC. When the isolated material was analyzed by ESI-MS, the m/z was 279.234. Further analysis by Raman spectroscopy revealed a peak similar to linoleic acid. This substance also possessed antimicrobial properties equivalent to linoleic acid.

Valorisation of By-Products from Soybean (Glycine max (L.) Merr.) Processing.

In recent years, increased awareness of the health benefits associated with consuming soy-based foods, knowledge of milk-related allergies and a move towards more sustainable food production have led to an increase in the number of available soy-based products. The biggest producers in the world, the USA, South America and China, are from the Pacific region. This enormous production is accompanied by the accumulation of related by-products, in particular, a substance that is known as okara. Okara is a paste that is rich in fibre (50%), protein (25%), fat (10%), vitamins and trace elements. Its proper use would lead to economic advantages and a reduction in the potential for polluting the environment. Its high fibre content and low production costs mean that it could also be used as a dietary supplement to prevent diabetes, obesity and hyperlipidaemia. Chemical or enzymatic treatment, fermentation, extrusion, high pressure and micronisation can all increase the soluble fibre content, and thus improve nutritional quality and processing properties. However, the product also degrades rapidly due to its high moisture content (70-80%), which makes it difficult to handle and expensive to dry by conventional means. The aim of this paper is therefore to thoroughly study the existing literature on this subject in order to develop a general protocol for okara exploitation and valorisation. A cost/benefit analysis could drive the design of eco-friendly, sustainable protocols for the preparation of high-value nutritional products.

Soy Products Ameliorate Obesity-Related Anthropometric Indicators in Overweight or Obese Asian and Non-Menopausal Women: A Meta-Analysis of Randomized Controlled Trials.

BACKGROUND: The effect of soy products on the weight of overweight or obese people is controversial, so we aimed to conduct a systematic review and a meta-analysis of published randomized controlled trials to analyze whether supplementation with soy products can help them to lose weight. METHODS: The relevant data before January 2019 in PubMed, Embase and Cochrane Library were searched. A random-effect model was adopted to calculate the weighted average difference of net changes of body weight, body mass index (BMI), body fat percentage, fat mass, waist circumference, etc. Results: A total of 22 trials (870 overweight or obese participants) were reflected in the present meta-analysis. Analysis showed that soy products significantly reduced body weight, BMI, body fat percent and waist circumference in overweight or obese Asian populations (-0.37 kg, P = 0.010; -0.27 kg/m2, P = 0.042; -0.36%, P = 0.032; -0.35 cm, P = 0.049) and more significant effects were observed in non-menopausal women reduced body weight (-0.59 kg, P = 0.041), BMI (-0.59, P = 0.041) and waist circumference (-0.59 cm, P = 0.041) in overweight or obese populations. CONCLUSION: This meta-analysis showed that soy products have weight loss effects, mainly due to soy protein, isoflavone and soy fiber.

New Evaluation of Isoflavone Exposure in the French Population.

The study relates the present evaluation of exposure to estrogenic isoflavones of French consumers through two approaches: (1) identification of the isoflavone sources in the French food offering, (2) a consumption-survey on premenopausal women. For the foodstuff approach 150 food-items were analysed for genistein and daidzein. Additionally, 12,707 labels of processed-foods from French supermarket websites and a restaurant-supplier website were screened, and 1616 foodstuffs of interest were retained. The sources of phytoestrogens considered were soy, pea, broad bean and lupine. A price analysis was performed. A total of 270 premenopausal women from the French metropolitan territory were interviewed for their global diet habits and soy consumption and perception. In supermarkets, there were significantly less selected foodstuffs containing soy than in restaurant (11.76% vs. 25.71%, p < 0.01). There was significantly more soy in low price-foodstuff in supermarket (p < 0.01). Isoflavone levels ranged from 81 to 123,871 µg per portion of the analyzed soy containing foodstuff. Among the women inquired 46.3% claimed to have soy regularly. Isoflavone intake >45 mg/day is associated to vegan-diet (p < 0.01). In total, 11.9% of soy-consumers had a calculated isoflavone intake >50 mg/day. This dose can lengthen the menstrual cycles. The actual exposure to phytoestrogen is likely to have an effect in a part of the French population.

Application of the slope-ratio growth assay technique to estimate tryptophan availability in soybean meal fed to young rats.

The slope-ratio assay for rat was used to determine whether tryptophan (Trp) availability in soybean meal (SBM) is affected by the presence of other amino acids (AAs). In a preliminary study, rats were fed graded levels of Trp-supplemented diets to establish the Trp concentration range over which the weight gain response was linear. This range was found to be from 0.04% to 0.12% Trp. Subsequently, rats were fed basal (0.045% Trp) or Trp-supplemented diets from three different sources: l-Trp alone, SBM, or l-Trp mixed with other AAs to reflect AA levels in the test SBM (AA-mix). Weight gain in rats increased linearly with supplemental Trp intake (p < .05) for all Trp sources. Compared to the slope achieved with l-Trp alone, the estimated availability of Trp in SBM was 84.4%, while for the AA-mix it was 93.4%. It is evident that the 6.6% reduction in l-Trp availability in AA-mix is due to metabolic costs derived from excess levels of other AAs beside Trp, given that the absorption of crystalline l-Trp in the small intestine is 100%. In conclusion, the Trp availability of SBM was estimated to be around 90.4% (i.e., 84.4/93.4 × 100) after correcting for the effects of the other AAs in SBM.

Toxicological evaluation of lotus, ginkgo, and garlic tailored fermented Korean soybean paste (Doenjang) for biogenic amines, aflatoxins, and microbial hazards.

The present study aimed to develop a consortium of nutritive fermented food products, supplemented with phytochemicals, with reduced toxicological contents. We developed new flavored Doenjang products (protein rich) fermented with lotus, ginkgo, and garlic plant extract-based Meju (termed as EMD) as the starter culture and by using traditional Meju (termed as TMD), where these plant extracts were added later during the fermentation process. Fermented Doenjang samples were analyzed for reduced levels of biogenic amines (BAs), aflatoxins, and microbial hazards, (including Bacillus cereus) as well as for their nutritive contents and antioxidant potential, after varying periods of fermentation (0, 3, 6, 9 and 12 months). All Doenjang samples prepared using plant extracts and their mixtures (1% and 10%) showed desired reduction in B. cereus counts, BAs, aflatoxins, and other foodborne pathogens as well as showed potent antioxidant abilities, including phenolic/flavonoid contents. Based on the higher efficiency in reducing various toxicants, Ginkgo biloba leaf extract added TMD samples were selected for the development of Doenjang products as an innovative approach, with great potential to improve the quality and safety of soybean fermented products in the Korean market, offering enhanced health benefits and reduced risks of toxicity.

The effects of nattokinase supplementation on collagen-epinephrine closure time, prothrombin time and activated partial thromboplastin time in nondiabetic and hypercholesterolemic subjects.

The aim of this study was to investigate whether supplementation with nattokinase, which is considered one of the most active functional ingredients found in natto, alters hemostatic factors. Subjects presenting with hypercholesterolemia (serum cholesterol: 200-280 mg dL-1) were randomly divided into nattokinase and placebo groups (n = 50, respectively). No significant between-group differences were found at baseline in collagen-epinephrine closure time (C-EPI CT), prothrombin time (PT), or activated partial thromboplastin time (aPTT). After 8 weeks of treatment, the nattokinase group exhibited significant increases in C-EPI CT, PT, and aPTT. The nattokinase group showed significantly greater increases in C-EPI CT (P = 0.001) and aPTT (P = 0.016) than the placebo group. Moreover, at 8 weeks, the nattokinase group showed a significantly higher C-EPI CT than the placebo group (P = 0.001). Additionally, a significant correlation between PT and aPTT was observed (r = 0.491, P < 0.001). In conclusion, nattokinase supplementation was associated with prolonged C-EPI CT and aPTT in nondiabetic and borderline-to-moderate hypercholesterolemic subjects.

Safety evaluation of Semen Sojae Preparatum based on simultaneous LC-ESI-MS/MS quantification of aflatoxin B1 , B2 , G1 , G2 and M1.

Semen Sojae Preparatum (SSP) is one of the most widely used traditional Chinese medicines, and is also a functional food. However, contamination with aflatoxins may occur in the fermentation process. To evaluate its safety, an accurate and rapid LC-ESI-MS/MS analytical method was developed and validated for the simultaneous determination of AFB1 , AFB2 , AFG1 , AFG2 and AFM1 in SSP. After a simple ultrasonic extraction of SSP samples, chromatographic separation was achieved on an Agilent Zorbax SB-C18 column (2.1 × 50 mm, 3.5 µm) with a flow rate of 0.50 mL/min. The gradient elution program was performed using a mobile phase consisting of water and acetonitrile, both containing 0.1% formic acid. Detection of five aflatoxins was based on triple quadrupole mass spectrometry using a multiple reaction monitoring mode with an electrospray ionization source. SSP is likely to be contaminated by aflatoxins in the processes of fermentation, storage, transportation and usage, and it is necessary to strictly monitor it. Artemisia annua L. and Morus alba L. may inhibit the production and growth of AFB1 - and AFB2 -producing fungi, which has a certain detoxification effect on contamination with aflatoxins in SSP.

Identification and characterization of multifunctional cationic peptides from traditional Japanese fermented soybean Natto extracts.

In this study, we investigated the lipopolysaccharide (LPS)-neutralizing and angiogenic activities of cationic peptides derived from the traditional Japanese fermented product Natto, which is made by fermenting cooked soybeans using Bacillus subtilis. Initially, we prepared 20 fractions of Natto extracts with various isoelectric points (pI's) using ampholyte-free isoelectric focusing (autofocusing). Cationic peptides were then purified from fractions 19 and 20, whose pH values were greater than 12, using reversed-phase high-performance liquid chromatography, and were identified using matrix-assisted laser/desorption ionization-time-of-flight mass spectroscopy. Among the 13 identified cationic peptides, seven (KFNKYGR, FPFPRPPHQK, GQSSRPQDRHQK, QRFDQRSPQ, ERQFPFPRPPHQK, GEIPRPRPRPQHPE, and EQPRPIPFPRPQPR) had pI's greater than 9.5, positive net charges, and differing molecular weights. These peptides were then chemically synthesized and applied to chromogenic LPS-neutralizing assays using Limulus amebocyte lysates, and 50% effective (neutralizing) concentrations of 2.6-5.5 µM were demonstrated. In addition, tube formation assays in human umbilical vein endothelial cells revealed angiogenic activities for all but one (GEIPRPRPRPQHPE) of these seven cationic peptides, with increases in relative tube lengths of 23-31% in the presence of peptides at 10 µM. Subsequent experiments showed negligible hemolytic activity of these peptides at concentrations of up to 500 µM in mammalian red blood cells. Collectively, these data demonstrate that six cationic peptides from Natto extracts, with the exception of GEIPRPRPRPQHPE, have LPS-neutralizing and angiogenic activities but do not induce hemolysis.

Fermented Ganjangs (Soy Sauce and Sesame Sauce) Attenuates Colonic Carcinogenesis in Azoxymethane/Dextran Sodium Sulfate-Treated C57BL/6J Mice.

The effects of different ganjangs (also designated as kanjang), including acid-hydrolyzed soy sauce (AHSS), fermented soy sauce (FSS), and fermented sesame sauce (FSeS), on azoxymethane (AOM, 10 mg/kg)/dextran sulfate sodium (DSS, 2%)-induced colorectal carcinogenesis in C57BL/6J mice were studied. Low doses (4 mL/kg) of both FSeS and FSS significantly increased colon length, suppressed AOM/DSS-induced increases in colon weight/length ratios, and induced colorectal neoplasia compared with AHSS-treated and control mice. Fermented sauces, particularly low doses of FSeS and FSS, showed activity against AOM/DSS-induced colorectal carcinogenesis by abrogating serum and mRNA levels of tumor necrosis factor-α, interferon-γ, interleukin (IL)-6, and IL-17α as well as by reducing mRNA levels of inducible nitric oxide synthase and cyclooxygenase-2 in colon mucosa. FSeS significantly increased colonic p53 expression compared with other sauces. However, AHSS showed weak activity against AOM/DSS-induced colonic carcinogenesis. Overall, FSeS showed the strongest anticancer effect, followed by FSS and AHSS. Thus, fermentation with microorganisms rather than chemical processes is important, and raw materials are another factor influencing anticancer activity.

Phytoestrogens in soy-based meat substitutes: Comparison of different extraction methods for the subsequent analysis by liquid chromatography-tandem mass spectrometry.

This paper presents the development of an efficient extraction procedure followed by a high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for the determination of five phytoestrogens (genistein, daidzein, formononetin, biochanin A, and coumestrol) in soy-based meat substitutes. Phytoestrogens are considered endocrine disrupting compounds, and their quantification is important in soy-based products, whose diffusion is increasing nowadays. The HPLC-MS/MS method, with electrospray ionization (ESI) source, was optimized to obtain high specificity and sensitivity, as well as rapidity of the analysis. Three extraction techniques were applied to soy burgers and compared: ultrasound assisted extraction, ultrasound assisted extraction followed by solid phase extraction and the Quick Easy Cheap Effective Rugged and Safe (QuEChERS) methodology. Both ultrasound assisted extraction and QuEChERS proved to be suitable for the determination of phytoestrogens, showing high recoveries, in the range of 86% to 99% and 75% to 105%, respectively. Matrix effect was evaluated, and ion suppression was observed for coumestrol and formononetin, demonstrating the importance of matrix effect assessment when complex samples are analyzed by HPLC-ESI-MS. The complete analytical protocols provided limits of detection and quantitation in soy-burgers at the ng g-1 level for all the considered phytoestrogens. Some soy burger samples were analyzed by both ultrasound assisted extraction and QuEChERS followed by HPLC-MS/MS. High concentration levels of daidzein and genistein (2-59 µg g-1 and 2-72 µg g-1 , respectively) were found; formononetin was in the range of 5 to 26 ng g-1 , while biochanin A and coumestrol were under the limit of quantitation in all samples. The results obtained with the two different sample treatment were in good agreement, proving the precision and accuracy of the described techniques.