Protein binding study of isoflavone, perillyl alcohol and S-ibuprofen by high-performance frontal analysis

High-performance frontal analysis (HPFA) was used for a protein binding study of isoflavones (daidzein, genistin, and genistein), enantiomers of perillyl alcohol and S-ibuprofen to human serum albumin (HSA). The analyses were performed on a Develosil and Inertsil 100-Diol-5 column (10 cm×4.6mm). Sodium phosphate solution (pH 7.4, ionic strength 0.17) was used as the mobile phase at a flow rate of 1 ml/min. To ensure the drug to be eluted as a trapezoidal peak with a plateau, injection volumes were each fixed up the zonal profile with an evident plateau appears. The unbound drug concentration was determined from a plateau height of the plateau region after that experimental data were fitted by Scatchard equation. The binding constants (K) and total binding affinities (nK) of drugs to HSA were calculated, respectively.     

Characterization of Fractionated Soy Proteins Produced by a New Simplified Procedure

It was possible to fractionate soy protein into two soy protein isolate fractions (>90% protein) enriched in either glycinin or β-conglycinin by using a new simplified procedure (referred to as the Deak procedure) employing CaCl₂ and NaHSO₃. The Deak procedure produced fractions with higher yields of solids, protein, and isoflavones, and similar protein purities as well as improved functional properties compared to fractions recovered by established, more complex soy protein fractionation procedures. The Deak glycinin-rich fraction comprised 15.5% of the solids, 24.4% of the protein, and 20.5% of the isoflavones in the starting soy flour, whereas the glycinin-rich fraction of the established procedure (Wu procedure) comprised only 11.6% of the solids, 22.3% of the protein, and 9.6% of the isoflavones. The Deak β-conglycinin-rich fraction comprised 23.1% of the solids, 37.1% of the protein, and 37.5% of the isoflavones in the starting soy flour, whereas the Wu β-conglycinin-rich fraction comprised only 11.5% of the solids, 18.5% of the protein, and 3.3% of the isoflavones. Protein purities were >80% for both fractions when using both procedures. The Wu procedure produced protein fractions with slightly higher solubilities and similar surface hydrophobicities; whereas, the fractions produced by the Deak procedure had superior emulsification and foaming properties and similar dynamic viscosity behaviors.     

Identification and determination of isoflavones in germinated black soybean sprouts by UHPLC−Q-TOF-MS mass spectrometry and HPLC-DAD

Ultra-performance liquid chromatography coupled with electrospray ionization tandem quadrupole time-of-flight mass spectrometry (UHPLC?Q-TOF-MS) and HPLC-DAD with an ultrasonic extraction method was developed for qualitative and quantitative compositions and contents in black soybean sprouts. Operational conditions of ultrasonic extraction were optimized by single-factor experiment. Phytochemical compositions of black soybean sprouts were identified by UHPLC?Q-TOF-MS in positive ion mode. A total of 23 compounds were tentatively characterized and identified by means of accurate mass and characteristic fragment ions. Among of them, the six isoflavones in different germinated black soybean sprouts were further quantified by HPLC-DAD. The results indicated that the calibration curves showed good linearity (r² > 0.9994). The intra-day and inter-day precision variations were less than 1.65% and 1.73%, respectively. The recoveries were in the range of 94.95–106.45%. The results indicated that the maximum amounts of isoflavones were produced on the fourth to fifth day germinated black soybean sprouts.     

大豆异黄酮酸水解条件的研究

研究了乙醇-水体系提取大豆异黄酮后,再经酸水解的最优工艺条件,经正交实验,确定最佳条件为原料与HC1比为1∶6,HC1浓度0.1mol/L,水解温度85℃;水解时间为1h。     

Effects of oven-drying,roasting, and explosive puffing process on isoflavone distributions in soybeans

Distributions of isoflavones in soybeans treated with oven-drying, roasting, or explosive puffing were analysed using high-performance liquid chromatography (HPLC). As oven-drying time increased from 0 to 120 min at 100 °C, concentration (μmol/g) of malonyl derivatives of isoflavones decreased and β-glucosides increased significantly with over 0.99 coefficient of determination (R²) (P < 0.05). Roasting at 200 °C for 7, 14, and 21 min and explosive puffing at 490, 588, and 686 kPa decreased malonyl derivatives significantly and increased acetyl-β-glucosides and β-glucosides significantly (P < 0.05). Total isoflavones (TI) in 21 min roasted and 686 kPa puffed soybeans decreased by 25.46% and 10.42%, respectively, while TI in 120 min oven-dried soybeans was not significantly different (P > 0.05) compared to untreated samples. Regression analysis showed that malonyl-β-genistin had higher slopes of decreases (μmol/g/min) than malonyl-β-daidzin in oven-dried soybeans. This is the first report on the effects of explosive puffing and the changes of isoflavone profiles in soybeans.     

大豆异黄酮抗大鼠T细胞衰老及抗氧化作用研究

为研究大豆异黄酮抗大鼠T细胞衰老和抗氧化作用，选用老龄SD大鼠，观察大豆异黄酮对T细胞CD28表达水平及肝脏氧化应激水平的影响。实验设立青年对照组、老龄对照组和大豆异黄酮0．33、0．99g／kgBW剂量组，其中剂量组按剂量灌胃给予大豆异黄酮，对照组灌胃给予等量纯净水。4周后，分别制备肝细胞匀浆和肝单细胞悬液，用硫代巴比妥酸（TBA）比色法测定肝匀浆中丙二醛（MDA）含量，分光光度法测定超氧化物歧化酶（SOD）活力，用流式细胞仪结合双氢罗丹明123（Rh123）和双氢-乙酰乙酸二氯荧光黄（82DCFDA）染色法分别检测肝细胞线粒体膜电位（MMP）及活性氧（ROS）水平；另取外周血测定T淋巴细胞表面CD8／CD28表达情况。结果显示：青年对照组及大豆异黄酮剂量组的肝组织MDA含量均较老年对照组低，而SOD活力及肝细胞MMP水平均较老年对照组高（P〈0．05）；大豆异黄酮剂量组肝细胞内的ROS水平较老年对照组低（P〈0．01）。老年对照组大鼠外周血CD28^＋和CD28^＋／CD8^＋T细胞比例低于青年对照组，而CD28^-／CD8^＋T细胞比例高于青年对照组，给予大豆异黄酮样品组三者比例均介于两对照组之间。提示衰老大鼠外周血T细胞CD28表达降低，肝细胞线粒体膜稳定性下降，氧自由基清除能力下降，过氧化脂质分解的终产物增多；大豆异黄酮可提高老龄大鼠T细胞CD28表达水平，增加肝细胞抗氧化酶活性，稳定线粒体膜电位，减少活性氧产生。     

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.     

Analysis of isoflavone concentration and composition in soybean [<Emphasis Type="Italic">Glycine max</Emphasis> (L.)] seeds between the cropping year and storage for 3 years

The objective of this study was to examine the relation between storage period and isoflavone concentration and composition in soybean [Glycine max (L.) Merrill] seeds. Total isoflavones varied from 470.0 to 878.1 µg g^–1 over the three cropping years and there were significant differences between the soybean cultivars. Generally, the malonyl conjugates were in higher concentrations than were the other compounds, and the individual isoflavones varied with each cropping year. The Myeongjunamul (1998: 936.7 µg g^–1, 1999: 727.0 µg g^–1, 2000: 807.8 µg g^–1) cultivar had the highest concentration, on average. Regarding changes in isoflavones with storage period, the concentration decreased by 91.7 to 171.1 µg g^–1. Malonyldaidzin and malonylgenistin concentrations also decreased, and the concentration of glucosides increased slightly over the 3 years. Acetyldaidzin, acetylglycitin and glycitein could not be detected or were only found in very small concentrations. Pureun (–66.2 µg g^–1) and Myeongjunamul (–77.4 µg g^–1) cultivars showed little variation in isoflavone levels, whereas the concentration in Muhan (–295.4 µg g^–1) decreased markedly on average over the three storage years. There were also significant differences between the cultivars. Our study suggests that it might be feasible to improve the storage methods of soybeans to preserve them for longer periods, which would have benefits for both their import and their export.S.H. Kim and W.S. Jung contributed equally to this work.     

Isoflavone profile of a high intensity pulsed electric field or thermally treated fruit juice-soymilk beverage stored under refrigeration

A fruit juice-soymilk (FJ-SM) beverage was treated by high intensity pulsed electric fields (HIPEF) (35 kV/cm with 4 μs bipolar pulses at 200 Hz for 800 or 1400 μs) or heated (90 °C, 60 s) in order to evaluate and compare the effects of both technologies on the isoflavone profile along the storage at 4 °C. Total isoflavone content was determined by quantification of aglycone and glucoside forms analyzed with HPLC. Immediately after treatments, all the evaluated samples showed a higher concentration of glucosides than aglycones, genistin being the most abundant glucoside. Heat treated samples showed higher initial concentration of glucosidic forms as compared to the untreated or HIPEF treated beverages. No significant changes were observed in total isoflavone content after the increasing HIPEF treatment time from 800 to 1400 μs. During the storage, genistein, daidzein and daidzin content increased; while genistin showed a slight decrease, irrespective of the treatment applied. In this way, total isoflavone content tend to increase throughout the time in all the evaluated samples. After 56 days of storage, there was no significant difference in HIPEF treated and fresh FJ-SM beverages, the latter being slightly higher in those thermally treated. As a result, HIPEF could be a good technology in order to obtain FJ-SM beverages with a high content of isoflavones and fresh-like characteristics. Considering the great concentration of isoflavones in untreated and treated FJ-SM beverages, their consumption could be a good way to increase the isoflavone daily intake of the population.Industrial relevanceHIPEF processing could be a competent technology in order to obtain FJ-SM beverages with fresh-like appearance and similar isoflavone content than just made products. The fruit juice-soymilk beverage evaluated in the present research has a great isoflavone content regarding to commercial soy based beverages. Its consumption could increase the current isoflavone intake of the population. Industry could apply this non-thermal technology in order to meet current consumers' claims for quality foods with high nutritional values.     

Intermolecular interactions in phytochemical model systems studied by NMR diffusion measurements

Novel functional foods, such as tomato juice with soy, represent a new strategy to increase consumption of health promoting ingredients and phytochemicals. Interactions between soy protein, isoflavones, and tomato carotenoids could impact the bioaccessibility and bioavailability of individual phytochemicals. The primary objective of this study was to assess possible interactions between daidzein and daidzin, soy protein and carotenoids using proton one-dimensional and two-dimensional pulsed field gradient nuclear magnetic resonance spectroscopy experiments.