黑豆皮花色苷的中压液相色谱制备及结构鉴定

以黑豆皮为实验材料,用乙醇浸提法对黑豆皮中的花色苷进行提取,用大孔吸附树脂对花色苷进行纯化,经冷冻干燥得到黑豆皮花色苷粗品。利用中压制备色谱对花色苷组分进行分离,通过质谱分析鉴定经中压制备色谱分离后的花色苷组分。结果表明:黑豆皮花色苷粗品中的总花色苷含量为26.9%,经中压制备色谱对花色苷粗品进行分离后的2峰中矢车菊素-3-葡萄糖苷纯度达到91.46%。黑豆皮中的主要花色苷为天竺葵素-3-O-芸香糖苷、芍药色素-3-O-葡萄糖苷、矢车菊素-3-O-葡萄糖苷和锦葵素-3-葡萄糖苷-4-乙醛。     

基于液相色谱-质谱联用技术定性定量分析黑豆种皮中可溶型和结合型花青素

以超声波辅助有机溶剂提取法获得黑豆种皮可溶型花青素提取物,再进一步对不含可溶型花青素的黑豆种皮残渣使用酸水解和碱水解以及酸碱/碱酸轮提水解,获得黑豆种皮结合型花青素提取物。采用超高效液相色谱-电喷雾-四极杆飞行时间质谱联用仪分析鉴定各黑豆种皮提取物中所含有的共17 种花青素成分,包括11 种花青素糖苷类：飞燕草素-3-O-葡萄糖苷、飞燕草素-3-O-半乳糖苷、矮牵牛花素-3-O-葡萄糖苷、矮牵牛花素-3-O-半乳糖苷、天竺葵素-3-O-芸香糖苷、矢车菊素-3-O-葡萄糖苷、矢车菊素-3-O-半乳糖苷、天竺葵素-3-O-己糖苷、芍药花素-3-O-己糖苷、天竺葵素-3-O-(6”-丙二酰葡萄糖苷)和芹菜定-3-O-(6”-丙二酰葡萄糖苷);6 种花青素苷元：飞燕草素、矢车菊素、矮牵牛花素、天竺葵素、芹菜定和芍药花素。采用高效液相色谱-电喷雾-三重四极杆质谱联用仪精确定量各类黑豆种皮提取物中的花青素含量,结果表明,酸性结合型花青素提取物中结合型花青素的总含量最高。此外,在黑豆种皮的可溶型花青素提取物中,花青素主要以花青素糖苷类形式存在,苷元含量相对极少;而在结合型花青素提取物中,则主要以花青素苷元为主,糖苷类化合物相对少见。     

热处理与大豆异黄酮苷元的转化分析

目的研究不同加热处理后大豆异黄酮苷元的含量和比例变化情况。方法大豆样品经烘箱50、100和150℃烘干,微波加热5min和炒熟等热处理后,由80%乙醇溶液超声提取,经高效液相色谱SB-C18柱(4.6mm×250mm,5μm)分离,0.2%冰乙酸+甲醇溶液梯度洗脱,紫外检测器260nm检测苷元和β-葡萄糖苷型大豆异黄酮含量。结果黄豆中检测出黄豆苷元、染料木素2种苷元和黄豆苷、黄豆黄苷2种β-葡萄糖苷。随烘箱加热温度升高,黄豆苷元含量增加1～5倍;染料木素增加3～15倍。炒豆中苷元和β-葡萄糖苷增加量最多。微波加热与50℃烘箱加热结果基本相同。青豆、黑豆与黄豆结果相近。结论加热使豆粉中部分糖苷型大豆异黄酮分解转化为苷元,活性成分增多,营养保健价值提高。     

紫玉米苞叶花色苷的纯化鉴定及热稳定性分析

为明确紫玉米苞叶花色苷的组成及比较单体花色苷的热稳定性,实验采用大孔树脂、凝胶树脂分离纯化紫玉米苞叶花色苷提取物,得到三个不同花色苷组分,并采用液质联机的方法鉴定其结构;将制备的花色苷提取物及三个单体组分进行热重仪器分析,比较不同组分的热稳定性结果表明紫玉米苞叶中共含有6种花色苷,通过分离纯化后能够得到三个组分,第一个组分为芍药-3-O-葡萄糖苷和天竺葵-3-O-葡萄糖苷的混合物,第二个组分为矢车菊-3-O-葡萄糖苷,第三个组分为矢车菊-3-(6' 丙二酰-葡萄糖苷).其中总花色苷提取物的热稳定性最强,矢车菊-3-(6'丙二酰-葡萄糖苷)次之,芍药-3-O-葡萄糖苷和天竺葵-3-O-葡萄糖苷的混合物与矢车菊-3-O-葡萄糖苷热稳定性相当,均较以上组分弱.     

霉菌型豆豉和纳豆中异黄酮含量的比较研究

目的:比较纳豆和豆豉中异黄酮含量的组成变化。方法:利用超声波提取,采用高效液相色谱法测定,色谱柱填料为Hypersil ODS2(4.6 mm×250 mm,5μm),流动相为甲醇(A)-水(B)进行梯度洗脱,检测波长为255 nm。结果:豆豉与纳豆相比较,豆豉中大豆苷、黄豆黄苷、染料木苷、大豆甙元、黄豆黄素、染料木素的含量分别为11,未检出,25,320,65,380μg/g;纳豆中中大豆苷、黄豆黄苷、染料木苷、大豆甙元、黄豆黄素、染料木素的含量分别为330,73,410,12,3.3,28μg/g。结论:豆豉中甙元的含量高,纳豆中糖苷的含量高。     

不同种植地区蓝莓果中花色苷的分布

目的:探究不同种植地区的蓝莓果实中花色苷含量与海拔高度、纬度等环境因素之间的关系。方法:选取不同海拔高度和纬度的10个种植地区“灿烂”品种的兔眼蓝莓果实为研究对象,采用高效液相色谱法(HPLC)和高效液相色谱-电喷雾质谱(HPLC-ESI-MS)定性和定量分析蓝莓果中花色苷的组成成分,比较不同种植地区的蓝莓果中花色苷苷元、糖基组成与含量的差异。结果:从蓝莓果中检测到5类花青素苷元,共13种花色苷,苷元含量组成由高到低为锦葵色素>矢车菊素>飞燕草素>矮牵牛素>芍药素;糖基组成由高到低为半乳糖苷>阿拉伯糖苷>葡萄糖苷,其中从芍药素中仅检测到半乳糖苷。蓝莓果中锦葵色素-3-O-半乳糖苷和矢车菊素-3-O-半乳糖苷的含量最高,而飞燕草素-3-O-葡萄糖苷和矮牵牛素-3-O-葡萄糖苷含量低,在部分地区的蓝莓果中缺失或未检测到。不同种植地区蓝莓果中花色苷组成虽基本一致但含量存在差异。云南龙朋代表的低纬度、高海拔地区种植的蓝莓果中花色苷总含量最高,高纬度、低海拔地区的浙江草塔的蓝莓果中花色苷总含量最低。结论:不同种植地区的地理环境影响蓝莓果中花色苷的分布,高海拔地区的环境条件更利于蓝莓果实中花色苷的合成和积累。     

固相萃取-液相质谱联用测定大豆中异黄酮类化合物的方法研究

<正>大豆异黄酮是一种植物化学素，属植物黄酮类，主要来源于豆科植物的荚豆类，在大豆中含量较高。研究发现，大豆异黄酮具有抑制肿瘤生长、降低心血管疾病、预防和治疗骨质疏松以及缓解妇女更年期综合症等多种生理保健作用。大豆中天然存在的大豆异黄酮共有12种，可以分为3类，即黄豆苷类、染料木苷类、黄豆黄素苷类，每类都以游离型、葡萄糖苷型、乙酰基葡萄糖苷型、丙二酰基葡萄糖苷型这4种形式存在。目前，如何简单准确地测试大豆中异黄酮的含量仍是检测领域一个有待探索的问题，     

桑葚花色苷的分离纯化及其热降解动力学研究

通过AB-8大孔树脂、乙酸乙酯萃取和Toyopearl TSK HW-40S凝胶柱层析对桑葚汁中的花色苷进行分离纯化,得到两个单一的化合物组分。经HPLC-MS鉴定这两种花色苷分别为矢车菊素-3-芸香糖苷和矢车菊素-3-葡萄糖苷。在此基础上,研究了不同纯度花色苷的降解动力学,结果表明:桑葚花色苷粗提物中所含的黄酮类化合物对花色苷的热降解有较强的保护作用,含黄酮的花色苷体系对p H值的变化更为敏感。在p H 3.5时,矢车菊素-3-芸香糖苷和矢车菊素-3-葡萄糖苷的热稳定性基本相同;在p H 4.0时,矢车菊素-3-芸香糖苷的热稳定性略好于矢车菊素-3-葡萄糖苷。     

蒸汽爆破对豆渣中大豆异黄酮的影响研究

采用高效液相色谱法研究蒸汽爆破处理对豆渣中大豆异黄酮组成和含量的影响。结果表明,豆渣经蒸汽爆破处理后,6种大豆异黄酮含量均显著增高,大豆苷、大豆苷元、黄豆黄苷、黄豆黄素、染料木苷、染料木素分别由汽爆前的3.69、11.22、0.84、2.76、2.13、27.45μg/g增至38.04、74.96、15.75、23.36、20.36、66.28μg/g。黄豆黄苷增幅最大,较汽爆前增高了17.75倍;染料木素增幅最小,较汽爆前增高了1.41倍。而且,大豆异黄酮的增幅随汽爆强度增加呈上升趋势,在汽爆压强2.0 MPa、维压时间30 s时达到最大值(黄豆黄苷为2.0 MPa、60 s)。研究表明,采用适宜的汽爆强度处理豆渣,能够显著提高大豆异黄酮含量,这为豆渣的开发利用提供了有益参考。     

自筛β-葡萄糖苷酶高产菌-N_1制备大豆异黄酮苷元的研究

目的:探索一种用自筛的β-葡萄糖苷酶高产菌-N1制备大豆异黄酮苷元的方法。方法:应用酶学研究常规方法在实验室条件下,用自筛的β-葡萄糖苷酶高产菌-N1获得β-葡萄糖苷酶液,探索制备大豆异黄酮苷元的工艺。结果:实验室制备的大豆异黄酮苷元样品纯度为91.33%,收率为11.44%。大豆异黄酮粗品经处理后,苷的数量减少,大豆苷和黄豆黄苷从28.21%降低到22.93%,染料木苷从8.24%降低到5.67%。苷元的数量明显的增加,大豆苷元从0.04%增加至0.05%,黄豆黄素从的0.003%增加至0.09%,染料木素从0.006%增加至0.07%。结论:研究表明以自筛菌-N1作为酶促反应制备大豆异黄酮苷元的β-葡萄糖苷酶产生菌完全可行。大豆异黄酮粗品经酶液处理后,苷元的数量有明显的增加,尤其是黄豆黄素和染料木素,分别增加了30倍和11.67倍。为进一步研究奠定了基础。     

不同品种大豆异黄酮组分及体外抗氧化活性分析

为研究不同品种大豆异黄酮组分和含量以及抗氧化活性,测定了50个特色大豆品种的百粒重、色泽（L*、a*、b*）,通过高效液相色谱法（HPLC）测定异黄酮组分,评估不同品种大豆甲醇提取物的DPPH、ABTS自由基清除能力,并用相关性分析、主成分分析及聚类分析法对样品进行分析。结果表明:大豆的百粒重范围为7.05~47.46 g。不同种皮色大豆L*、a*和b*呈现显著差异（P<0.05）。HPLC分析结果发现糖苷型异黄酮含量是大豆中主要的异黄酮组分,占总异黄酮含量的90%以上。其中染料木苷含量最高。安豆115品种的大豆异黄酮的总含量最高,为2500.78 μg/g;靖江丝瓜青品种的大豆异黄酮含量最低,为888.86 μg/g。安豆115品种的DPPH和ABTS自由基清除能力较强,分别为（16.11±0.25）和（8.12±0.04）μmol V_C/g。糖苷型异黄酮含量与DPPH和ABTS自由基清除能力有较高的线性相关性（R2分别为0.903和0.867）。主成分分析表明6项指标可用2个主成分来表示（累计贡献率达66.3%）。聚类分析将50个品种的大豆分为4类:第一类中黄豆黄素和黄豆黄苷含量较高;第二类中大豆苷、大豆苷元、染料木苷和染料木素含量较高;第三类和第四类中其大豆异黄酮含量基本处于中等水平和较低含量。综上所述,不同大豆品种的异黄酮组成、含量和抗氧化能力存在较大差异,可为其进一步综合开发利用提供一定理论基础。     

Soluble phenolics and antioxidant properties of soybean (Glycine max L.) cultivars with varying seed coat colours

This research was the first to investigate nutritional components, including soluble phenolics (isoflavones and anthocyanins), protein, oil, and fatty acid as well as antioxidant activities in different coloured seed coat soybeans (yellow, black, brown, and green) for two crop years. The soluble phenolics differed significantly with cultivars, crop years, and seed coat colours, while protein, oil, and fatty acid exhibited only slight variations. Especially, malonylgenistin and cyanidin-3-O-glucoside compositions had the most remarkable variations. Green soybeans had the highest average isoflavone content (3079.42 μg/g), followed by yellow (2393.41 μg/g), and black soybeans (2373.97 μg/g), with brown soybeans showing the lowest value (1821.82 μg/g). Anthocyanins showed only in black soybeans, with the average contents of the primary anthocyanins, cyanidin-3-O-glucoside, delphinidine-3-O-glucoside, and petunidin-3-O-glucoside, quantified at 11.046, 1.971, and 0.557 mg/g, respectively. Additionally, Nogchae of green soybean and Geomjeongkong 2 of black soybean may be recommended as potential cultivars owing to the highest average isoflavone (4411.10 μg/g) and anthocyanin (21.537 mg/g) contents. The scavenging activities of 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) radicals also differed remarkably, depending upon isoflavone and anthocyanin contents, with black soybeans exhibiting the highest antioxidant effects.     

不同大豆品种大豆皂苷组成分析

为明确山西大豆资源胚和子叶中大豆皂苷含量的分布情况,筛选特异资源在育种中应用,本研究以71份山西大豆种质资源为材料,采用高效液相色谱-电喷雾离子化串联质谱联用技术(HPLC-ESIMS/MS)分析不同材料中大豆皂苷组成和含量的差异。结果表明:参试材料胚和子叶中大豆皂苷含量变异范围分别为17.02~83.51 mg/g和6.53~50.11 mg/g。A类、DDMP类、B类和E类皂苷均与总皂苷含量呈极显著正相关。小粒型大豆材料胚中B类和E类皂苷含量、子叶总皂苷和DDMP类皂苷含量显著高于中粒和大粒型材料。野生、半野生大豆材料胚中B类、E类大豆皂苷含量显著高于栽培品种;野生大豆子叶总皂苷、DDMP类皂苷含量显著高于半野生和栽培大豆。     

Comparisons of phenolic compounds, isoflavones, antioxidant capacity and oxidative enzymes in yellow and black soybeans seed coat and dehulled bean

The total and individual phenolic profiles and their contribution to antioxidant capacities of seed coats and dehulled beans of black and yellow soybeans were investigated. In addition, lipoxygenase (LOX) and peroxidase (POD) activities were also determined. Black seed coats had the highest content of total phenols, flavonoids and anthocyanins, in addition the highest catechin and protocatechin contents. On the other hand, dehulled beans from yellow soybeans had the highest levels of ferulic and p-coumaric acid content, and free and bound isoflavone content. The LOX activity was concentrated in the dehulled beans of both black and yellow soybeans, while a high POD activity that can be attributed to a dominant gene EpEp was presented in seed coats of yellow soybean genotype Galeb. Due to the high concentration of phenols, the black seed coats from soybean genotypes Black Tokio and Cornaja had the highest ABTS^·+-scavenging capacity (597.46 and 486.15 mmol Trolox Eq./kg d.m., respectively). The results suggest that the black seed coat as well as dehulled bean from yellow soybeans would potentially provide sources of natural antioxidants that may play a crucial role in human health protection.     

Biological activities of cheonggukjang prepared with several soybean cultivars

To select proper soybean cultivars for producing functional cheonggukjang, a comparison was made of the physiological activities of different cheonggukjang prepared with 30 different soybean cultivars. The isoflavone content was highest in the cheonggukjang made from ‘Daepung’ soybeans at 208.75 mg%. In general, the contents of glycone types (and derivatives) of isoflavone, specifically daidzin, glycitin, genistin, and malonylgenistin, were higher than that of aglycone types. The polyphenol contents ranged from 30.62 to 80.32 mg%. The DPPH radical scavenging activity and superoxide dismutase (SOD)-like activity of the cheonggukjang made from yellow soybeans had a higher activity than those of black soybeans. Although there are no consistent tendencies in the functional activity of cheonggukjang according to soybean color and size, the antioxidative activity is highest in the cheonggukjang made of yellow soybeans. Additionally, the fibrinolytic and inhibitory activities against angiotensin I-converting enzyme are highest in the cheonggukjang made of black soybeans. From these results, it can be concluded that the DPPH free radical scavenging activity and SOD-like activity in cheonggukjang depends on the phenolic compound content in soybean.     

A correlation between the level of phenolic compounds and the antioxidant capacity in cooked-with-rice and vegetable soybean (Glycine max L.) varieties

This study was undertaken to determine the content of phenolic compounds contained in embryo, cotyledon and seed coat of nine soybean (Glycine max L.) varieties. The 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activity was also measured to determine the correlation between the level of phenolic compounds and antioxidant capacity. A total of 10 anthocyanin constituents and 21 phenolic compounds was detected and quantified. In all nine varieties, the seed coat and cotyledon had the highest and lowest levels of phenolic compounds and anthocyanins, respectively despite the fact that all of them showed a wide variation in total amounts of phenolic compounds and anthocyanins in seed coat, embryo, and cotyledon. The seed coat tissue, but not other seed parts, showed a strong correlation between the seed coat color and the content of both phenolic compounds and anthocyanins. The brown and black soybean seed coat contained much higher levels of phenolic compounds and anthocyanins in seed coat tissue than the yellow or green coat soybean. Among the individual phenolic compounds, syringic acid (214 μg g⁻¹) and chlorogenic acid (31 μg g⁻¹) were highest in seed coat and embryo, respectively. Myricetin was highest both in whole seed (16.7 μg g⁻¹) and cotyledon (16.0 μg g⁻¹), being equivalent to 20 and 30% of total phenolic compounds, respectively. Among the 10 anthocyanins, cyanidin-3-glucoside was found to accumulate at the highest level in the seed coat (1783 μg g⁻¹), whole seed (106 μg g⁻¹) and embryo (0.35 μg g⁻¹), which correspond to 95, 96, and 40% of the total anthocyanin contents, respectively. The cotyledon accumulated pelargonidin-3-glucoside (0.39 μg g⁻¹) at the highest level that is equivalent to 62% of total anthocyanin contents. DPPH activity was found to have a strong correlation (^***probability < 0.001) with phenolic compounds (0.67^***) and anthocyanins (0.70^***).J. A. Kim, W. S. Jung, and S. C. Chun are equally contributed to this work.     

Hydrolysis of soybean isoflavone glycosides by a thermostable β-glucosidase from Paecilomycesthermophila

The β-glucosidase from Paecilomyces thermophila J18 was found to be capable of hydrolysing daidzin and genistin in a previous study. This report further evaluated the thermostability and hydrolysis of soybean isoflavone glycosides. The enzyme was found to be very stable at 50 °C, and retained more than 95% of its initial activity after 8 h at 50 °C. It converted isoflavone glycosides, in soybean flour extract and soybean embryo extract, to their aglycones, resulting in more than 93% of hydrolysis of three isoflavone glycosides (namely, daidzin, genistin and glycitin) after 4 h of incubation. Also, addition of the β-glucosidase greatly increased the contents of isoflavone aglycones in the suspended soybean flour and soymilk. The results indicate that the thermostable β-glucosidase may be used to increase the isoflavone aglycones in soy products. This is the first report on the potential application of fungal β-glucosidases for converting isoflavone glycosides to their aglycones in soy products.     

Effect of soybean varieties on the content and composition of isoflavone in rice-koji miso

To clarify the effect of soybean varieties on isoflavone, a useful component for human health, in soybean products, we investigated changes in the isoflavone content and composition in rice-koji miso, after fermentation/aging for 6 or 12 months using varieties of soybeans (Tohoku-126, Tohoku-135, Tohoku-139, Suzuyutaka and Chinese soybeans), by high performance liquid chromatography. In soybeans, the total isoflavone content in Tohoku-126 was 444 mg/100 g, which was 1.2–2.0 times the content in the other soybean varieties. The malonyl glycosides and aglycones in soybeans accounted for more than 60% and only a few percent, respectively. As for rice-koji miso, the total isoflavone and aglycone contents were the highest in miso prepared from Tohoku-126. The ratios of glycosides to aglycones (80.1–92.6%) in miso were higher than those in the original soybeans. The time course of the isoflavone composition during the fermentation/aging process of rice-koji miso indicated that glycosides decreased from 86.4% to 44.9% after 6 months but aglycones increased from 9.6% to 53.3%.     

Chemical,rheological and sensory characteristics of sweet spreads made from by-products of soya bean and maize

Sweet maize cob (SMC) and black soya bean seed coat (BSC) were used to prepare a sweet spread. Chemical composition of the prepared spread and its rheological and sensory characteristics were investigated. The results were compared with those of commercial plum and raspberry jams and spreads. Spreads made from SMC and BSC had the highest content of proteins, total fibres, hemicellulose and cellulose. In relation to fruit jams, they are distinguished by the presence and content of p-coumaric, ferulic and 3,4-dihydroxybenzoic acids, as well as of flavonoids such as catechin and quercetin. SMC spread with 3.2% of BSC had the highest content of total anthocyanins (666.75 mg CGE kg⁻¹) (cyanidin 3-glucoside equivalent). Cyanidin-3-glucoside (Cy-3-Glu) was the most abundant anthocyanins in SMC/BSC containing spreads, while in raspberry jam, that was cyanidin-3-sophoroside (Cy-3-Sop). According to chemical, rheological and sensory properties, SMC/BSC containing spreads could be competitive with fruit jams and spreads.