微生物发酵产大豆异黄酮苷元的连续超声波提取工艺及其定量分析研究

选取浅玫瑰链霉菌、红色糖多孢菌及黑曲霉为发酵菌种,采用Plackett-Burman设计及正交试验,对影响微生物发酵法产大豆异黄酮苷元(染料木黄酮、黄豆苷元、黄豆黄素)的连续超声波提取工艺的主要试验因素进行系统优化。以浅玫瑰链霉菌为发酵菌种的实验结果表明：各主要试验因素对3种异黄酮苷元提取率影响程度大小为：抽提溶剂组分、超声波时间、淋洗液体积。而温度、溶剂流速两个因素与提取效率负相关。连续超声波作用时间及抽提溶剂组分为影响异黄酮苷元提取工艺的两个主要影响因素。当两因素的最优值分别为7min及70%时,总异黄酮苷元提取率为(98.7±1.23)%。通过固相萃取(SPE)可有效去除发酵提取液中杂质对HPLC定量分析的干扰,并使3种异黄酮苷元含量浓缩10倍以上,从而使HPLC方法的检测限达到10-3μg/mL,样品总分析时间小于10min,样品回收率为96.2%～103.4%,RSD值小于2.34%。上述结果表明,该方法可满足微生物发酵法产大豆异黄酮苷元的高效提取纯化及快速定量分析。     

氧化石墨烯的制备及其催化水解大豆异黄酮

采用改良的Hummers法制备了负载有-SO3H、-COOH和-OH的氧化石墨烯,对其进行了结构表征。并首次以氧化石墨烯为催化剂,对天然产物大豆异黄酮糖苷水解反应进行了研究。实验结果显示氧化石墨烯在异黄酮糖苷水解反应中具有较好的催化活性,其最佳反应温度为105℃,此时3种大豆异黄酮糖苷的转化率分别达到94.3%,92.1%和88.8%,苷元的收率分别达到69.6%、60.6%和58.8%。其催化活性明显优于其他固体酸催化剂,如沸石HZSM-5和大孔树脂NKA-9,而与0.02 mol.L.1的硫酸相当。     

Ripening temperature affects the content and distribution of isoflavones in sufu,a fermented soybean curd

Sufu, a fermented soybean curd, was prepared by ripening the salted tofu cubes in the Aspergillus oryzae‐fermented rice–soybean koji mash at 25, 35 or 45 °C for a period of 16 days. It showed that the 16‐day ripened sufu contains less total isoflavone content (629.29–739.68 μg g^?1 dried matter) than the salted tofu before ripening (942.59 μg g^?1 dried matter). Regardless of ripening temperature, ripening causes a major reduction in the content of β‐glucoside and malonylglucoside isoflavones along with a significant increase of aglycone isoflavone content. These changes were enhanced as the ripening period extended. Among the various treatments examined, sufu ripened at 45 °C showed the greatest increase in aglycone content coupled with the greatest decrease in malonylglucosides. The distribution of malonylglucosides decreased from an initial 40.32% in the tofu cube to 9.78% after 16 days of ripening at 45 °C. Meanwhile, the distribution of aglycone increased from 13.17% to 39.88%.     

乙醇/磷酸氢二钾双水相体系分离大豆异黄酮

利用乙醇/磷酸氢二钾双水相体系浸提大豆异黄酮固体,并采用四因素四水平正交试验,考察了乙醇质量分数、磷酸氢二钾质量分数、温度、时间对大豆异黄酮的分配系数和萃取收率的影响。结果发现,影响大豆异黄酮萃取收率的因素为:时间磷酸氢二钾质量分数乙醇质量分数温度。最佳提取条件为:25%乙醇/25%磷酸氢二钾双水相体系,室温(20℃)浸提30 min。大豆异黄酮几乎全部分配在上相,其上相收率为97.0%。     

Antioxidant Properties of Genistein in a Model Edible Oil System

ABSTRACT:  Antioxidant properties of genistein were tested during the low temperature oxidation of linseed oil. Four linseed oil solutions were prepared: linseed oil, linseed oil containing 2 μmol added genistein/g oil, linseed oil containing 4 μmol added genistein/g oil, and linseed oil containing 200 ppm butylated hydroxyanisole (BHA). Dimethyl sulfoxide (DMSO) (1.7%) was added to all solutions to allow for the solubilization of genistein. Antioxidant activities were analyzed using 2 techniques: (i) the oven stability test carried out at 60 °C with periodic determination of peroxide values (PV); and (ii) oxidation on a heated (60 °C) FTIR-ATR crystal with periodic scanning and collection of infrared spectra. Induction points (IP) for each solution were determined. A strong correlation ( r = 0.91) was noted between the 2 analytical methods used. Three significantly ( P < 0.10) different levels of IP were generally observed. The shortest IP (17 h for PV, 36.9 h for FTIR) was found for the linseed oil. Intermediate IP's were obtained for the 2 μmol added genistein/g oil (32.4 h for PV, 43.5 h for FTIR) and 4 μmol added genistein/g oil (33.9 h for PV, 44.2 h for FTIR) samples. The longest IP (39.9 h for PV, 54.2 h for FTIR) was obtained for the 200 ppm BHA containing solution. Although genestein was not as effective as BHT in retarding oxidation, the results clearly demonstrate that it is able to act as an antioxidant in a bulk oil system.     

大豆低温脱溶粕中两种异黄酮单体分离与结构鉴定

以大孔树脂柱色谱、制备薄层色谱从大豆粕乙醇提取物中分离出二种异黄酮单体,经HPLC,UV,IR,ESI／MS,和1H NMR,13C NMR,DEPT NMR,HQSC NMR分析确证其结构是大豆甙(daidzin)和染料木素(genistein)。     

酸水解法提取大豆异黄酮甙元工艺研究

以脱脂大豆粕为原料，采取酸水解后用无水乙醚萃取的方法提取大豆异黄酮甙元成分并通过单因素实验和正交实验确定了最佳提取工艺参数。实验结果表明，酸水解法提取大豆异黄酮的提取率和产品纯度均高于常规有机溶剂浸提法。     

Influence of the dehydration process on active compounds of okara during its fractionation

Okara (residue of the soymilk manufacture) is rich in proteins, fibres and lipids. It also contains isoflavones that possess health‐promoting properties. A new method has been developed for the valorization of fibres from okara by hydrolysis of insoluble proteins with a protease and removal of the oil. Three different processes were investigated: the first one involved delipidation and drying prior to proteolysis and led to the highest content of fibre (80%) in the final product. The second used proteolysis on crude okara followed by solvent delipidation–dehydration and gave an intermediate content of fibre (75%). The last process was totally enzymatic (proteolysis and lipolysis) and gave the lowest content of fibre in the final product (50%). Fibre water‐holding capacity was correlated to the total dietary fibre content of each sample. It was preferable to use crude okara for hydrolysis, since oven‐drying during the process decreased the water‐holding capacity and modified the isoflavone profile of okara. Copyright © 2005 Society of Chemical Industry     

Modeling of Nanofiltration Process for Solvent Recovery from Aqueous Ethanol Solution of Soybean Isoflavones

Abstract

In this paper, the feasibility of recovering the solvent from the aqueous ethanol solution of soybean isoflavones with nanofiltration (NF) was studied. Five commercially available polymeric NF membranes were employed and STARMEM? 122 showed acceptable flux and high retention. The central composite design (CCD) of the response surface methodology (RSM) was applied to model the effects of temperature, pressure, and feed concentration on the permeate flux and the total soybean isoflavone retention. The results indicate that the developed models were in good agreement with the experimental results and they can be used to predict this NF process.     

Assessment of the estrogenicity of the diet of a healthy female Spanish population based on its isoflavone content

Phytoestrogens are natural components of plants consumed by humans. The hormonal activity of these substances has long been known. Various in vitro tests have demonstrated the agonistic or antagonistic estrogenic activity of some phytoestrogens. The objective of this study was to estimate the supply of isoflavones in the diet of a healthy adult female population and to assess its estrogenic effect. The diet was assessed by questionnaire and the estrogenicity of the estimated isoflavone content was tested by E-screen, finding a mean total estrogenic capacity of 0.129 × 10^?10 eq.E₂ (12.9 pmol day^–1), corresponding to a daily isoflavone intake of 265.8 μg day^–1. This study offers a preliminary insight into the phytoestrogen content of the diet of a healthy active population of Spanish women. The effects of this additional hormonal burden are highly controversial, and this approach to estimating dietary phytoestrogen intake of specific populations may help to elucidate its implications for human health.