High-power ultrasonication-assisted extraction of soybean isoflavones and effect of toasting

The effect of high-power ultrasonication (HPU) on extraction of soybean isoflavones and components, including total phenolic and DPPH radical scavenging activities (RSA) were evaluated. Isoflavones from defatted soybean flakes were extracted using a bench scale HPU system at 20 kHz and varying amplitudes (18–54 μm) for 1 and 3 min. Aqueous acetonitrile, and aqueous ethanol based solvents were evaluated for extraction at samples-to-solvent ratios of 0.1:1 and 0.2:1. The non-sonication control extraction was with mixing with magnetic stirrer for 2 h. Preliminary data indicated 1.2–1.5 times more genistein recovery at 1, and 3 min sonication compared to control, however, total phenolic decreased. The genistein recovery was lower at higher sonication levels. RSA also decreased to 40% for HPU-assisted extractions. Total isoflavones recovery in HPU-assisted extractions increased from 600 to 5813 μg/g. The concentration of major isoflavone components, genistein, daidzein, and glycitein also increased by 10-fold. HPU for 3 min reduced the average particle size of treated soy flakes from 530 μm to 60 μm. Toasting at 150 °C of defatted soy flake for longer period of time led to higher aglycone concentration in extract; however, 2 h toasting was sufficient prior to HPU-assisted extraction.     

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.     

Functional attributes of soybean seeds and products,with reference to isoflavone content and antioxidant activity

The concentrations of isoflavones, especially daidzein and genistein, were determined by high performance liquid chromatography in 4 soybean cultivars and 26 soybean products. The total isoflavone content of the soybean cultivars was in the range of 525–986 mg per kg, and for soy products it was 32.9–795 mg per kg. Amongst the soybean products, the isoflavone content decreased in the order: soy sprouts, soy seeds, soy flour, soy milk, soy meals and soy sauce. Significant differences in the concentration of genistein and daidzein were observed between the commercial soy products and also within the soybean cultivars. The antioxidant activity of soybean and soy products correlated well with total phenolic content (TPC) and total isoflavones (TI), whereas TPC showed higher correlation with TI.     

Soybean Isoflavones: Effects of Processing and Health Benefits

Isoflavones belong to a group of compounds known as flavanoids that share a basic structure consisting of two benzene rings linked through a heterocyclic pyrane C ring. Soybean provides the most abundant source of isoflavones, which have been linked to the prevention of several diseases, including cancer (prostate, colon, and breast cancer), hypercholesterolemia, cardiovascular disease, atherosclerosis, osteoporosis, and relief of menopausal symptoms in certain women. Currently, isoflavone use is targeted as a nutraceutical. In this article, we review the structural aspects and the effect of processing on the different forms of isoflavones in soy protein. We also discuss the impact of these changes on the health benefits of eating foods containing isoflavones. Isoflavones, as a class of phytoestrogens, are gaining a lot of interest in clinical nutrition.     

酱油渣异黄酮抗氧化功效成分纯化研究

以酱油渣为原料,采用乙醇提取法提取酱油渣中大豆异黄酮,对提取条件进行了优化。比较了醇沉法、等电点沉淀法、浓缩离心法、正丁醇法、二氯甲烷法、乙酸乙酯萃取法等6种方法对异黄酮粗提物的纯化效果。通过高效液相鉴定了纯化产物的异黄酮单体组成,并采用邻苯三酚氧化法和DPPH法评价了异黄酮纯化物的体外抗氧化活性。结果表明：优化条件下提取的异黄酮粗提物中异黄酮质量分数为1.21%、提取率为（85.36±0.09）%。6种纯化方法中,浓缩离心法所得异黄酮产物中异黄酮纯度为7.75%,去除蛋白质效果最佳;二氯甲烷萃取法所得异黄酮产物中异黄酮纯度为27.74%,去除总糖和灰分效果最佳;两者联用后所得纯化产物的异黄酮纯度为44.88%,其主要由染料木素、大豆黄素和大豆素组成,质量分数分别为67.37%、9.33%、23.30%。在质量浓度为1 mg/mL时,异黄酮纯化物对超氧阴离子、DPPH自由基的清除率分别为67.65%、89.18%,显著高于异黄酮粗提物（P＜0.05）,其IC50分别为0.34、0.26 mg/mL,说明异黄酮纯化物具有一定的抗氧化活性,有望作为一种潜在的天然抗氧化剂应用于功能性食品中。     

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.     

Daidzein and genestein contents in tempeh and selected soy products

The total isoflavones in tempeh and selected local soy products was determined. Raw tempeh contained 26 ± 6 mg daidzein (Da) and 28 ± 11 mg genestein (Ge) while fried tempeh contained 35 ± 11 mg Da and 31 ± 11 mg Ge in 100 g (wet basis). Total isoflavone content in 100 g of raw tempeh, based on a dry weight, was 205 ± 56 mg and significantly reduced to 113 ± 41 mg in 100 g of fried tempeh. Tempeh in batter was deep-fried for 30 min which reduced 45% of the total isoflavone content compared to the raw one. Raw tempeh contained the highest total content of isoflavone among the studied local soy products. Total isoflavone content in processed soy foods like egg tofu and home made soybean drink were significantly lower than other soy products studied.     

Introduction and nutritional evaluation of germinated soy germ

Germinated soy germ (GSG) were developed and evaluated for their nutritional value. Separated soy germ was germinated at room temperature for 24 h under running water. As germination progressed, the protein and fibre content of GSG increased slightly, while the lipid and carbon to nitrogen (C/N) ratio decreased; free amino acids including GABA increased considerably while free sugars decreased. Linoleic and linolenic acid were the most abundant unsaturated fatty acids in soy germ, and slight changes were observed in GSG. The tocopherol and isoflavone contents showed a rapid increase of 32.4% and 27.9%, respectively, during germination. The abundance of GABA, isoflavones and tocopherols demonstrates the high nutritional value of GSG and suggests that GSG can be utilised as a reasonable and effective source of healthy foods.     

酱油渣中大豆异黄酮的提取研究进展

异黄酮是一类具有重要生物活性的化合物,在大豆和大豆制品中含量十分丰富。本文综述了国内外近年来对酱油渣中大豆异黄酮的提取研究概况,主要包括各种提取方法及其优缺点以及在工业生产过程中的应用前景,以期为大豆异黄酮作为保健食品进一步开发应用奠定一定基础。     

Effect of fermentation by pure and mixed cultures of Streptococcus thermophilus and Lactobacillus helveticus on isoflavone and B-vitamin content of a fermented soy beverage

Enhancement of nutritional or vitamin content of foods is commonly touted as a major benefit of probiotics. In this paper, we examined the ability of three probiotic bacteria either alone or in combination to enhance nutritional content. Pure and mixed cultures of Streptococcus thermophilus ST5 and Lactobacillus helveticus R0052 or Bifidobacterium longum R0175 were used to prepare fermented soy beverages. The effects of strain, acidification and mixed cultures on the deconjugation of soy isoflavones were examined. Acidification to pH 4.7 alone resulted in a 7% drop in isoflavone levels. Deconjugation levels varied between the different glucosides. Fermentation by L. helveticus R0052 resulted in the 50% reduction in total glucosides with O-malonyl glucosides being reduced 64%. Fermentation by S. thermophilus ST5 or B. longum R0175 had no significant effect on isoflavone levels. Combining a S. thermophilus strain with a L. helveticus culture reduced the effectiveness of the latter. Fermentation did not significantly modify vitamin B1 or B6 levels.     

豆粕中大豆异黄酮的超声辅助提取工艺研究

以大豆异黄酮提取率为指标,豆粕为原料,采用超声波辅助提取大豆异黄酮。在单因素的基础上,通过正交试验确定提取豆粕中大豆异黄酮的最佳工艺条件。结果表明,从豆粕中提取大豆异黄酮的最佳工艺条件为:料液比为1∶20,乙醇浓度为80%,超声时间为30 min,超声温度为60℃。在此工艺条件下,大豆异黄酮的平均提取率为0.3522%。     

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.     

Profiling and quantification of isoflavones in soymilk from soy protein isolate using extracted ion chromatography and positive ion fragmentation techniques

Dietary supplements on soy based foods and beverages are increasingly gaining prominence all over the world. In this study, liquid chromatography coupled with positive electrospray ionisation tandem mass spectrometry (LC-ESI-MS) and diode array detection was used for the quantitation and characterisation of isoflavones in fermented and unfermented soymilk made from soy protein isolate SUPRO 590. Bifidobacterium animalis ssp. lactis Bb12 was used for the fermentation of soymilk. The isoflavones were found to produce characteristic radical ions as well as molecules of H₂O, CO₂, a sugar unit, and an alcohol through collision-induced fragmentation. Product ion fragments revealed unique fragmentation pathways for each isoflavone compound. Characteristic fragmentation of different isoflavones were unequivocally identified and differentiated. The occurrence of aldehydes such as pentanal, ethanal and methanal was shown to be specifically linked with isoflavone aglycones, daidzein, genistein and glycitein, respectively. Main glycosides such as genistin, daidzin and glycitin as well as the acetyl-, and malonyl forms also showed respective aglycone ions in their spectra fragmentation. Thus positive ion fragmentation was important in the absolute confirmation of isoflavones and to reveal the occurrence of other related compounds such as aldehydes in soymilk.     

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.     

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.     

Intake of isoflavone-supplemented soy yogurt fermented with <Emphasis Type="Italic">Enterococcus faecium</Emphasis> lowers serum total cholesterol and non-HDL cholesterol of hypercholesterolemic rats

The aim of this study was to obtain an isoflavone-supplemented soy yogurt, fermented with Enterococcus faecium CRL 183 and Lactobacillus helveticus ssp jugurti, with suitable sensory properties and to assess the effects of the final product on blood lipids in hypercholesterolemic rats. Four isoflavone supplementation procedures were tested, in which the isoflavone was added at these stages: (1) before heat-treatment; (2) after heating and before fermentation; (3) after fermentation and (4) in the okara (by-product of soy milk) flour stirred into the fermented product when consumed. The products were subjected to a test of sensory acceptability. To assess their potential hypocholesterolemic properties in vivo, four groups of rats were used: control (C), hypercholesterolemic (H), hypercholesterolemic plus fermented product (HF) and hypercholesterolemic plus isoflavone-supplemented fermented product (HFI). Hypercholesterolemia was induced in rats of groups H, HF and HFI by feeding them on a commercial rat chow to which cholesterol and cholic acid had been added. Total, HDL and non-HDL cholesterol and triglycerides were measured in the blood of the rats. No significant sensorial differences were detected among the samples of soy yogurt supplemented with isoflavones at various processing stages. Rats fed a fermented soy product enriched with isoflavones (HFI group) had significantly (P < 0.05) less serum total cholesterol (15.5%) compared with rats fed a hypercholesterolemic diet (H group). Non-HDL cholesterol was less (P < 0.05) in rats fed a fermented soy product enriched or not with isoflavones (27.4 and 23.2%) compared to H group. The HDL-C and triglyceride concentrations did not differ significantly among the groups. It was possible to obtain an isoflavone-supplemented soy yogurt with satisfactory sensory characteristics. The resulting supplemented soy yogurt was capable of producing a lipid-lowering effect in hypercholesterolemic rats, relative to the animals that did not consume this product.     

不同大豆制品中大豆异黄酮含量的比较研究

大豆异黄酮是一类具有重要生物活性的化合物,在大豆及其制品中含量丰富。本实验采用乙醇热回流提取法研究大豆及其制品中异黄酮含量。结果表明:样品加标回收率大于99%,说明该方法适用于大豆样品异黄酮含量的测定;大豆及其制品中异黄酮含量的大小顺序依次为:大豆>豆浆>豆腐脑>卤水豆腐>腐竹>白豆干>豆腐皮>油豆腐>脆豆皮,表明加工工艺会影响大豆异黄酮的含量。     

Preparation of isoflavones enriched soy protein isolate from defatted soy hypocotyls by supercritical CO2

Soy hypocotyls, the byproduct from soy protein industry, rich in isoflavones, was attempted to develop into a value-added functional food, soy protein isolate (SPI). Soy hypocotyls were defatted by supercritical CO₂ (SC-CO₂) under the following conditions as 35 MPa, 2 h, 45 °C with a constant flow rate of CO₂ at 5 l/min, fed from the bottom of the extraction vessel. Via extraction, the oil contents were lowered from 10.5 g/100 g in original soy hypocotyls to 0.2 g/100 g in defatted soy hypocotyls (DSH) with a Protein Dispersibility Index (PDI) value of 91.2 that contained about 2.18 g/100 g of isoflavones and 63.6 g/100 g of proteins. SPI was then extracted from DSH under the optimized conditions including: the pH of water solution at 6, the ratio of DSH to water solution as 1:10 and the extraction temperature at 30 °C with the extraction time of 30 min. The freeze-dried SPI contained 92.46 g/100 g of the proteins and 640 mg/100 g of isoflavones, where the three major β-glycosides, glycitin, daidzin and genistin accounted for about 83.7% of the total isoflavones. The yields of SPI and isoflavones were about 48% and 30% in terms of their respective total contents in DSH.     

Tracking isoflavones: From soybean to soy flour,soy protein isolates to functional soy bread

Soybean seeds with three different levels (low, intermediate and high) of isoflavones were processed to soy flour and soy protein isolates (SPIs) and developed into functional soy breads. The effect of factors involved in all steps of the process was investigated by tracking the composition and concentration of native forms of isoflavones. The total isoflavone contents were 8033.3, 10570.1 and 15169.0 nmol/g DM (dry matter) in the three soybeans; 13201.5, 20034.4 and 26014.3 nmol/g DM in defatted soy flours; 9113.2, 13274.6 and 17918.3 nmol/g DM in the SPI; 2782.7, 4081.4 and 5590.3 nmol/g DM in soy breads, respectively. The bread making processes did not affect the total isoflavone content, but changed glucosides/acetylglucosides to aglycones. Malonylglucosides were stable prior to baking but degraded to acetylglucosides and further to glucosides during baking. Our results provide critical information for the production of functional soy breads that contain varying amounts of soy isoflavones.