Characterization of Isoflavones in Membrane-processed Soy Protein Concentrate

ABSTRACT: Aqueous soy flour solutions (5% w/w) were treated for 3 h at 45 to 50 °C with Crystalzyme®, without enzyme at 45 to 50 °C, or blanched at 80 °C. Solutions were ultra‐ and dia‐filtered to produce membrane soy concentrates (MSC). Total isoflavones of MSCs were 3.02 mg/g, 3.12 mg/g, or 3.42 mg/g, respectively, on a dry weight basis. Membrane processing contributed to approximately 18, 15, or 8% decreases, respectively, in total isoflavones of MSCs compared with soy flour (3.71mg/g). This represents at least an 82% recovery. There was no significant difference in total isoflavone content in any MSC, however the profile of isoflavones as glucosides or aglycones changed with treatment conditions.     

Isoflavone profiles,phenol content,and antioxidant activity of soybean seeds as influenced by cultivar and growing location in Ohio

Isoflavone levels and isoflavone chemical composition in soybeans vary between planting locations although the exact factors which control isoflavone biosynthesis are unclear. We compared levels of 12 isoflavones in soybean seeds of six cultivars grown in four different locations in Ohio in 2002 as determined by high‐performance liquid chromatography. Antioxidant activity contained in plant‐based foods can improve food oxidative stability and phenolics and isoflavones have proven active in food systems. Radical scavenging activity was assessed using the 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH) radical. Total phenolics (TPCs) were determined by using Folin–Ciocalteu reagent. Total isoflavones (TIs) varied five‐fold (1573–7710 nmol g^?1) between seeds from the various location–cultivar combinations. One location (Wooster, Ohio) produced seeds containing half the isoflavones as the other locations tested apparently due to poor growing conditions. The cultivars could be divided into two groups based on TI, one having approximately 50% more isoflavones. Surprisingly, across the entire data set, with increasing TI, the proportion of isoflavones accounted for by the daidzein family increased due primarily to malonyl daidzin. DPPH scavenging did not differ significantly by location or cultivar (P > 0.05) and did not correlate with TPC or TI. Profiling soybean isoflavones could help elucidate how isoflavone biosynthesis is regulated and lead to better disease resistance of soybean crops and soy foods with greater health benefits. Copyright © 2007 Society of Chemical Industry     

Efficacy of isoflavones in relieving vasomotor menopausal symptoms – A systematic review

This review assessed the efficacy of isoflavone supplements to reduce vasomotor symptoms in menopausal women by reviewing all published randomized controlled trials. Systematic literature searches were carried out in 70 databases. Randomized and placebo controlled studies were included if they investigated the treatment of isoflavone supplements derived from soy or red clover on vasomotor symptoms in peri‐ or postmenopausal women for at least 12 wks. Data were analyzed concerning outcome and methodological quality of the study. Twenty‐three trials met the inclusion criteria, thereof 17 investigated soy isoflavones and 6 red clover isoflavones. Without exception, selected trials examining the effect of red clover isoflavones were already assessed in several meta‐analyses and were therefore excluded from this evaluation. As the soy isoflavone studies were very heterogeneous concerning interventions and outcome measures, meta‐analysis could not be performed and trials were systematically assessed in a structured approach. Included soy isoflavone studies had numerous quality deficiencies and did not consistently show a reduction of flushes after treatment with soy isoflavones. Therefore, there is no conclusive evidence, but only some indication of a benefit of soy isoflavones on hot flush frequency or severity.     

Urinary isoflavone excretion in Korean adults: comparisons of fermented soybean paste and unfermented soy flour

BACKGROUND: This study compared the 24 h urinary excretion of isoflavone algycones after the consumption of fermented soybean paste or unfermented soy flour in a population of healthy Koreans (four males and five females). An equivalent amount (20 mg) of total isoflavone was consumed as either 52 g of soybean paste or 12.2 g of soy flour in a randomised, crossover trial consisting of two single‐time‐point feedings and having 3 day run‐in and washout periods between feedings. RESULTS: Analysis of starting materials indicated that daidzein (1.4%) and genistein (1.4%) constituted a smaller proportion of the total isoflavones in soy flour as compared with soybean paste (55.3% daidzein and 45.7% genistein). Free and glycosidic forms of isoflavones differed significantly between the two soy products. Levels of β‐glycosides as well as total glycosides (β‐glycosides, malonyl and acetyl forms) were two‐fold higher in soy flour as compared with soybean paste. Malonyl forms predominated in soy flour. CONCLUSIONS: The consumption of soybean paste resulted in significantly higher urinary excretion of both daidzein and genistein as compared with soy flour (P < 0.05 and P < 0.01 respectively). Copyright © 2007 Society of Chemical Industry     

Effects of the β‐glycosidase reaction on bio‐conversion of isoflavones and quality during tofu processing

BACKGROUND: Isoflavones are the most common group of phytoestrogens which are present in significantly large amounts in soybean and soy products such as tofu. Isoflavones occur naturally in glycoside forms having lower bioavailability than their aglycone forms. β‐Glycosidase acts as a bio‐catalyst for the conversion of isoflavone glycosides to isoflavone aglycones, raising the bioavailability of isoflavones; therefore, it can be used to improve the quality of tofu. We need to establish process conditions for the optimal outcome of the enzyme reaction in tofu. RESULTS: By using the β‐glycosidase (0.02% w/v) reaction at 55 °C for 30 min, a maximum 84.5% conversion of isoflavone glycoside to isoflavone aglycone was obtained. The enzyme reaction caused no significant effects on the sensory acceptability of soft tofu. The hardness of enzyme‐treated hard tofu increased with the coagulant amount whereas prolonged heating resulted in decrease of hardness. Incorporation of enzyme reaction before the coagulation process during soft tofu processing provided a sufficient bio‐conversion of isoflavones at optimal conditions. CONCLUSION: β‐Glycosidase can be effectively used for the bioconversion of isoflavones in soft tofu manufacturing process at optimal reaction conditions before the onset of coagulation process. Copyright © 2010 Society of Chemical Industry     

INFLUENCE OF HEAT TREATMENT AND GRAIN GERMINATION ON THE ISOFLAVONE PROFILE OF SOY MILK

Soy foods have attracted much attention for their possible effects on human health because of their phytochemical content, mainly isoflavones. Twelve forms of isoflavones have been identified in soybeans and soy products. Technological processes like heat treatment or fermentation can change the isoflavone profile of soy products. In this study, the effects of heat treatment and grain germination on isoflavone profile of soy milk prepared from BRS 213 soybean variety were evaluated. Isoflavone forms were separated and quantified by reversed‐phase high‐performance liquid chromatography. Heat treatment by autoclaving for 5 min did not affect the total isoflavone content, but led to a 90% decrease in malonyl forms, whereas β‐glycosides increased by 70%. Autoclaving for 15 min led to a 20% loss of total isoflavones. Germination for 3 days did not change the total isoflavone content, but changed the distribution profile (15% increase of malonyl forms and 30% decrease of β‐glycosides). Germination for 7 days increased the bioavailability of aglycone isoflavones in soy milk by seven times.     

Dietary intake of isoflavones and polyunsaturated fatty acids associated with lung function,breathlessness and the prevalence of chronic obstructive pulmonary disease: Possible protective effect of traditional Japanese diet

The Japanese diet is high in soy products and fish. A case–control study was conducted in Japan to investigate the relationship between dietary intake of isoflavones and fatty acids and lung function, breathlessness and chronic obstructive pulmonary disease (COPD). A total of 278 referred patients aged 50–75 years with COPD diagnosed within the past 4 years, and 340 community‐based controls were assessed for respiratory symptoms and undertook spirometric measurements of lung function. A validated food frequency questionnaire was administered face‐to‐face to obtain information on habitual food consumption. Dietary intakes of isoflavones and fatty acids were derived from the Japanese food composition tables. The COPD patients had significantly lower habitual intakes of isoflavones (genistein and daidzein) and polyunsaturated fatty acids (PUFA; both omega‐3 and omega‐6) than control subjects. Lung function measures were found to be positively associated with isoflavones and PUFA intake. Substantial reductions in prevalence of COPD and breathlessness were observed for isoflavones, the respective adjusted odds ratio being 0.36 (95% confidence interval 0.19–0.68) and 0.60 (95% confidence interval 0.33–1.10) for the highest versus lowest levels of total isoflavone intake. The corresponding tests for linear trend were significant. High intakes of PUFA and omega‐6 fatty acids (derived from foods excluding oils and fats as seasonings) also appeared to reduce the risks of COPD and breathlessness symptom, but no evidence of association was found for other types of fatty acids. The study provided evidence of possible protective effect of traditional Japanese diet against tobacco carcinogens.     

Content and Profile of Isoflavones in Soy-Based Foods as a Function of the Production Process

Soy has been traditionally incorporated in diet as processed foods, such as soymilk, tofu, miso, tempeh, etc., and the consumption is commonly associated with a reduction of the development of chronic diseases due to their antioxidant, anti-inflammatory, and anti-allergic properties, among others. Many of the health benefits of soy have been attributed to isoflavones. They comprise a group of naturally occurring flavonoids consisting of heterocyclic phenols. Soy contains three types of isoflavones in four chemical forms: the aglycones daidzein, genistein, and glycitein; the β-glucosides daidzin, genistin, and glycitin; their 6″-O-malonyl-β-glucosides (6OMalGlc); and their 6″-O-acetyl-β-glucosides (6OAcGlc) conjugates. Industrial processing methods of soy-based food products commonly lead to the loss of isoflavones through removal of undesirable fractions. On the other hand, isoflavones can be transformed into different conjugates, which may have significant effects on the food texture and on the bioavailability and pharmacokinetics of the isoflavones. This article reviews the effect of a number of soybean processing treatments on the isoflavone content and profile. The preparation and manufacturing of different soy-based food and food ingredients, fermented and non-fermented, has been analyzed in terms of content and distribution of the three major isoflavone derivatives, daidzein, genistein, and glycitein, and their respective conjugates.     

Effect of free or protein-associated soy isoflavones on the antioxidant status in rats

BACKGROUND: The objective of this study was to investigate the effect of chronic ingestion of free and protein‐associated soy isoflavones on the antioxidant status in male Wistar rats. Free isoflavone (iso), protein‐associated soy isoflavone (iso + prot) and soy protein (prot) extracts were administered for 30 days by gavage to the rats at a dosage of 1 mg aglycone isoflavones per 200 g body weight, adjusted daily, and the prot group was given the same concentration of soy protein received by the iso + prot group. Antioxidant capacity of plasma, thiobarbituric acid‐reactive substance (TBARS) and glutathione (GSH) levels and catalase (CAT), glutathione peroxidase (GPx) and superoxide dismutase (SOD) activities in plasma, erythrocytes and tissues and gene expression levels in liver and kidney were evaluated. RESULTS: Chronic ingestion of free but not of protein‐associated soy isoflavones nor of solely soy protein increased plasma antioxidant capacity and GPx activity in erythrocytes. Soy protein increased CAT activity and gene expression in liver. SOD activity in erythrocytes was increased by all treatments. CONCLUSION: The overall results confirm that dietary soy isoflavones have a positive effect on antioxidant status, enhancing antioxidant capacity of plasma and antioxidant enzymes in various tissues, but the effects are dependent on the form of administration and on a complex mechanism of antioxidant status balance on the organism. Copyright © 2010 Society of Chemical Industry     

Modification of isoflavone profiles in a fermented soy food with almond powder

Isoflavone profiles of a fermented soy food, cheonggukjang, were modified using almond powder. Isoflavones were analyzed by high performance liquid chromatography (HPLC) with an ultraviolet detector. Malonyl derivatives of isoflavones decreased and aglycones of isoflavones increased in samples with almond powder for 48 h. As added, almond powder increased from 0%, 5%, and 10% (w/w), amounts of aglycones increased to 21.11%, 26.63%, and 32.45% for 48 h, respectively. β-Glucosidase activity in 5% and 10% almond added samples was significantly higher than samples without addition of almond (P < 0.05). The content of succinyl daidzin and succinyl genistin, new metabolites from isoflavones, in almond-added cheonggukjang was significantly lower than control samples, implying that β-glucosidase activity from almond affected negatively the formation of succinyl derivatives (P < 0.05). Principal component analysis (PCA) for isoflavone distribution showed that first principal component (PC1) and second principal component (PC2) expressed 64.78% and 22.26% of the data variability, respectively. Biotransformation of isoflavones in any fermented soy foods can be achieved using natural products containing high β-glucosidase activity such as almond. PRACTICAL APPLICATION: The results of this study can help to modify the structural transformation of phytochemicals in any fermented soy foods using natural products. Adjusting the content of almond powder can achieve wanted profiles, for example, high aglycones content. Also, content of metabolites such as succinyl derivatives can be controlled using proper amounts of almond and fermentation time.     

Soybean isoflavones: Efficacy of extraction conditions and effect of food type on extractability

Soy isoflavones are phytochemicals of intense interest due to their association with a variety of health protective effects. Analytical techniques to identify and quantify these compounds vary in accuracy, reproducibility and sensitivity. In this study, solvent extraction efficiencies, and the effect of replicate extractions, and sonication on the isoflavone content of two soy products (high and low protein content) were evaluated. The later study was conducted to determine the effect of protein content on isoflavone extractability. Repeated extractions (up to five sequential extractions) of soy meal sample increased the total concentration of isoflavones by 74%, 69% and 65%, using acetonitrile–HCl, methanol and ethanol. The increment for soy protein isolate was 147%, 103% and 105%, respectively. Sonication of both samples in the three solvents for 15 min extracted as much isoflavones as the total of the five sequential extractions. These results strongly indicates that, a one step extraction without sonication can markedly under-estimate the concentration of isoflavones in foods, and that protein content of foods may have a marked impact on isoflavone extractability.     

Isoflavone Profile in Soymilk as Affected by Soybean Variety,Grinding, and Heat‐Processing Methods

Isoflavones impart health benefits and their overall content and profile in foods are greatly influenced at each step during processing. In this study, 2 soybean varieties (Prosoy and black soybean) were processed with 3 different grinding (ambient, cold, and hot grinding) and heating methods (traditional stove cooking, 1‐phase UHT, and 2‐phase UHT) for soymilk making. The results showed after cold, ambient, and hot grinding, the total isoflavones were 3917, 5013, and 5949 nmol/g for Prosoy; the total isoflavones were 4073, 3966, and 4284 nmol/g for black soybean. Grinding could significantly increase isoflavone extraction. The grinding process had a destructive effect on isoflavones and this effect varied with grinding temperature. Different heating methods had different effects on different isoflavone forms. Two soybean varieties showed distinct patterns with respect to the change of isoflavone profile during processing.     

大豆异黄酮与肠道微生物相互作用研究进展

膳食摄入大豆异黄酮与人体健康有密切的关系。大豆异黄酮糖苷组分与肠道菌群相互作用可产生生物活性和生物可利用度显著提高的新型微生物转化物,促进大豆异黄酮生理活性充分发挥。同时,大豆异黄酮通过调控肠道菌群结构影响结肠中的微生物酶活性,改变结肠菌群代谢能力。本文将对大豆异黄酮与肠道微生物的相互作用进行综述,重点阐述肠道微生物对大豆异黄酮的生物转化作用、大豆异黄酮对肠道微生物及其酶活性的调控作用以及大豆异黄酮及其肠道转化产物对健康的作用。以期为膳食组分和肠道菌群相互作用促进人体健康研究领域提供理论参考。     

Effects of extraction temperature,ionic strength and contact time on efficiency of bis(2‐ethylhexyl) sodium sulfosuccinate (AOT) reverse micellar backward extraction of soy protein and isoflavones from soy flour

BACKGROUND: Soy protein enriched with isoflavones has been linked to various disease‐preventing and health‐promoting activities owing to the antihypertensive, hypocholesterolaemic, antiobesity and antioxidative properties of isoflavones. The isoflavone profiles of soy‐based products are known to be highly dependent on the various chemical and physical treatments to which the products have been subjected. The aim of this research was to increase the efficiency of backward extraction of soy protein and isoflavones from bis(2‐ethylhexyl) sodium sulfosuccinate (AOT) reverse micelles by studying the effects of extraction temperature, ionic strength of the aqueous stripping solution and contact time on the amounts of soy protein and isoflavones backward extracted from an AOT/H₂O/isooctane reverse micellar system. RESULTS: By modifying the extraction temperature, ionic strength and contact time, 47.0–60.2% of protein, 43.3–68.4% of daidzin, 43.8–74.6% of genistin, 39.0–88.8% of glycitin, 20.8–92.6% of malonyl genistin, 20.2–52.0% of malonyl glycitin, 32.7–75.6% of acetyl genistin, 49.7–76.8% of daidzein and 19.6–38.1% of genistein present in the AOT reverse micellar solution were backward extracted into the aqueous stripping phase. Statistical analysis showed that there were significant linear and interactive effects of temperature and contact time on the backward extraction of daidzin, genistin, glycitin and daidzein. Significant linear and interactive effects of ionic strength and contact time were found in the backward extraction of daidzin and genistin. The backward extraction of genistein was only influenced by contact time and its interaction with temperature. CONCLUSION: This study showed the potential of reverse micelles as a protocol for extracting isoflavones from soy samples for analytical purposes. By modifying the extraction temperature, contact time and ionic strength, soy protein enriched with daidzin, genistin, daidzein and genistein could be produced from soy flour. The results represent an important contribution to current knowledge on utilising reverse micellar extraction in food technology. Copyright © 2007 Society of Chemical Industry     

Evaluation of the composition and concentration of isoflavones in soy based supplements,health products and infant formulas

Isoflavones are weak estrogenic compounds found in soybeans and soy foods. The objective of the present study was to determine the content and composition of isoflavones in different soy isoflavone supplements, soy based health products and infant formulas. Commercially available samples of 13 different soy supplements and 11 soy based health products obtained from South East Asia were analyzed for their isoflavone concentrations by using high-performance liquid chromatography and their peak identities confirmed by LC–MS. The extent of daily intake of isoflavones from different products based on the directions for their consumption was further evaluated. The amount of isoflavones in dietary supplements, soy based health products and infant formulas ranged from 405 to 57,570, 46.32–1333.80 and 59.54–226.84 μg/g of the sample, respectively, expressed as aglycone equivalents.     

Physicochemical Characterization and Sensory Analysis of Yeast‐leavened and Sourdough Soy Breads

Sourdough fermentation has been shown to have numerous beneficial effects on bread quality, and nutritionally enhance soy‐supplemented bread by altering isoflavone chemical forms. Given this, the objective of this study was to compare the loaf quality and shelf life of sourdough and yeast‐leavened soy breads by various physical, thermal, and sensorial methods, and to assess the effects of fermentation by various microorganisms on isoflavone profile in dough and breads using high‐performance liquid chromatography analysis. Sourdough fermentation yielded a less extensible dough compared to yeast‐leavened soy dough (P < 0.001), and resulted in a harder bread crumb (P < 0.05) and lighter crust color (P < 0.001), compared to yeast‐leavened soy bread (Y‐B). Sensory analysis revealed a significantly higher overall liking of Y‐B compared to sourdough soy bread (SD‐B) (P < 0.001). Segmentation analysis of the cohort suggests that overall liking and bread consumption frequency may be determinants of Y‐B or SD‐B preference. SD‐B and Y‐B exhibited similar shelf‐life properties. Despite significantly different enthalpies associated with the melting of amylose‐lipid complexes, thermal analysis of the 2 soy breads stored for 10 d (ambient conditions) demonstrated no significant difference in water distribution and starch retrogradation (P < 0.05). Lastly, SD‐B was determined to have 32% of total isoflavones occurring in the aglycone form compared to 17% in Y‐B. These findings warrant further investigation of sourdough fermentation as a processing technique for quality and nutritional enhancement of soy‐based baked goods.     

Systematic evaluation of pre-HPLC sample processing methods on total and individual isoflavones in soybeans and soy products

There are mainly two protocols in isoflavone analysis, one that involves hydrolysis prior to HPLC analysis and the other direct HPLC analysis. In this study, three different hydrolysis methods were systematically re-evaluated, and compared with direct HPLC analysis. Acidic hydrolysis (1.2-3 M HCl in ethanol at 80 °C) showed a maximum conversion of ca. 92% from glucosides to aglycones in 2 h; however, longer reaction caused degradation of genistein. Alkaline hydrolysis using 2 M NaOH converted acetylglucosides and malonylglucosides to their respective glucosides within 10 min. Glucuronidase from H. pomatia effectively converted isoflavone glucosides and acetylglucosides to aglycones. Quantification of the total isoflavones in various soy food products showed no significant difference among direct injection and the three hydrolysis methods (P < 0.05). We conclude that direct analysis of isoflavones in crude extracts is a rapid and accurate method to obtain isoflavone profiles and compositions in soybean, soy foods and beverages.     

Stability and Shelf Life of Bioactive Compounds during Food Processing and Storage: Soy Isoflavones

ABSTRACT: During the past several years, the nutrition community has recognized the importance of the nonnutritive components of plant foods. These nonnutritive components are commonly called phytochemicals. One of the most exciting developments in the phytochemicals field is the beneficial biological activity of isoflavones from soy. Benefits include the potential reduced cancer risk, cholesterol homeostasis, and other hormone replacement therapy treatments. Yet, very little attention has been given to the interactions of bioactive phytochemicals with other food ingredients. Various studies demonstrated the loss of soy isoflavones during storage and the effect of processing conditions on specific modifications of soy isoflavones. Beyond apparent 1st-order degradation kinetics, there is a change in the isoflavones profile according to processing and storage conditions. This change in chemical structure is very likely to affect the bioavailability and bioactivity of soy isoflavones. Understanding the reactions that affect soy isoflavones bioactivity will provide a basis for developing food with the beneficial quality of isoflavones. This article presents current information on the stability of soy isoflavones and highlights how these changes may affect their bioavailability and biological activity. Most importantly, the aim of this article is to point out the fact that much more data are required to fully understand the chemistry and the kinetics of isoflavones reaction in model systems and food.     

从豆豉中提取大豆异黄酮及含量的测定

豆豉是一味中药，因其具有消食化滞、发汗解表、除烦平喘等疗效被我国卫生部定为第一批药食兼用品种。豆豉的生产起源于我国，但由于豆豉含盐量较高等原因一直被当成调味品而使用。其实豆豉中还含有大豆异黄酮等重要功能因子，文章采用单因素优选法制备了大豆异黄酮并进行了分析检测，豆豉中异黄酮含量为820ug／g，这对充分挖掘豆豉中异黄酮等活性成分、提高豆豉的知名度、扩大豆豉加工范围、开发保健食品新资源意义很大。