反相高效液相色谱法测定苦荞中的芦丁和槲皮素

目的:建立一种苦荞黄酮中芦丁和槲皮素含量的测定方法。方法:分别以芦丁和槲皮素为标品,采用KromasilC18反相柱(4.6mm×300mm,5μm),分别测定水解和未水解苦荞黄酮样品中芦丁和槲皮素的含量。结果:在流动相为0.4%磷酸-甲醇(体积比为40∶60)、检验波长358nm、进样速度1.0mL/min、柱温20℃条件下,测定的芦丁含量的线性范围为0.025～0.125g/L(r=0.9993),槲皮素含量的线性范围为0.008～0.256g/L(r=0.9995)。槲皮素的加样回收率99.9%,变异系数为1.50%(x,n=5);芦丁的加样回收率98.5%,变异系数为1.16%(x,n=5)。水解样中槲皮素含量为(194.7±4.22)mg/g,未水解样中槲皮素含量为(8.96±0.26)mg/g,芦丁含量为(83.84±3.29)mg/g,变异系数(x,n=3)分别为:2.17%、2.90%和3.92%。结论:该方法简便、快速、准确,可用于苦荞黄酮中芦丁和槲皮素的含量测定。     

苦荞萌发物中生物活性黄酮对人乳腺癌细胞增殖的抑制作用

以萌发期(1～6d)的苦荞芽粉为原料,采用铝盐显色可见分光光度法和HPLC 法,探明萌发期(1～6d)苦荞芽粉乙醇提取物中总黄酮、芦丁和槲皮素含量的变化趋势。以人乳腺癌细胞MCF-7 为模型,采用噻唑蓝(MTT)比色法,比较萌发期(1～6d)的苦荞芽粉乙醇提取物对人乳腺癌细胞体外的增殖抑制率。结果表明：苦荞芽粉乙醇提取物具有抑制MCF-7 乳腺癌细胞增殖的作用,尤以萌发第3 天(芦丁与槲皮素含量比为0.92:1)时抑制效果最好,显示二者具有良好的协同抑制效果;苦荞芽粉乙醇提取物的抑制效果与槲皮素和芦丁标准品模拟样品抑制效果相似,表明苦荞芽粉乙醇提取物对MCF-7 细胞的生长起抑制作用的主要功效成分为槲皮素和芦丁。     

不同苦荞茶中芦丁和槲皮素含量测定及二者冲泡溶出率比较研究

本文建立了HPLC法测定6款造粒成型苦荞茶、8款全麦(胚)苦荞茶中芦丁、槲皮素含量,并研究两类苦荞茶中芦丁、槲皮素的冲泡溶出率。采用Phenomenex C₁₈色谱柱(4.6 mm×250 mm,5 μm),以甲醇(A)-0.1%冰乙酸(B)为流动相梯度洗脱,流速1.0 mL·min-1,检测波长257 nm,柱温30 ℃。在此色谱条件下,芦丁和槲皮素完全分离,线性范围分别为0.00362~0.90613 mg·mL-1(r=0.9996)、0.01439~0.71947 mg·mL-1(r=0.9998),平均回收率分别为101.99%(RSD 1.71%)、98.37%(RSD 2.03%)。结果表明,造粒成型苦荞茶中芦丁和槲皮素总量17.34 mg·g-1,槲皮素占89.10%,其冲泡液中芦丁、槲皮素总溶出率19.41%;全麦(胚)苦荞茶中芦丁、槲皮素总量11.96 mg·g-1,芦丁占91.39%,其冲泡液中芦丁、槲皮素总溶出率67.86%。苦荞茶冲泡液中芦丁、槲皮素总量:造粒成型苦荞茶<全麦(胚)苦荞茶(P<0.01)。采用SPSS 22.0对14款苦荞茶及冲泡液中芦丁、槲皮素总量和溶出率进行聚类分析,可分为造粒成型苦荞茶和全麦(胚)苦荞茶两类。     

不同光质对苦荞芽黄酮类物质及抗氧化活性的影响

将苦荞分别在红光(630 nm)、蓝光(460 nm)、白光(540 nm)条件下发芽,研究其芦丁、槲皮素、花青素含量和抗氧化活性的变化。采用高效液相色谱法(HPLC)测定苦荞芽中芦丁、槲皮素、花青素含量,用分光光度法测定苦荞芽提取物对DPPH自由基、ABTS自由基的清除能力。结果表明,光照条件下苦荞芽的生长受到抑制,其中抑制效果为蓝光白光红光;在蓝光条件下,苦荞芽中芦丁、槲皮苷和槲皮素含量显著增加,同时其抗氧化活性有所提高;红光和蓝光条件下,苦荞芽中花青素能够有效累积。     

苦荞不同部位酚类化合物组成与抗氧化活性

目的:研究苦荞不同部位酚类化合物组成与抗氧化活性。方法:利用70%乙醇分别提取苦荞壳、苦荞麸皮和苦荞粉三部分中的酚类物质,并分析提取物的酚类化合物组成和抗氧化活性。结果:麸皮部位总酚、总黄酮分别为(3 042±31) mg GAE/100 g·DW和(5 290±233) mg RE/100 g·DW,分别占全苦荞的60.8%和67.7%。芦丁是麸皮提取物中的主要酚类化合物,其含量为(5 152±61) mg/100 g·DW。苦荞壳、苦荞麸皮和苦荞粉提取物均表现出抗氧化活性,其中苦荞麸皮提取物的抗氧化活性最强,其ORAC、FRAP、DPPH自由基清除率和ABTS自由基清除率分别为(1 503.3±249.5) μmol/g·DW、(8 477.5±441.1) mg TE/100 g·DW、(5 642.4±241.3) mg TE/100 g·DW和(16 764±742) mg TE/100 g·DW。结论:麸皮是苦荞酚类化合物的主要富集部位,这些化合物表现出较强的抗氧化活性。     

萌发荞麦中芦丁和槲皮素含量变化的研究

研究分析荞麦萌发过程中芦丁和槲皮素含量的变化趋势,为荞麦萌动食品的开发提供了理论参考。以发芽时间为试验因素,采用反相液相色谱法对荞麦萌发前后的芦丁和槲皮素进行测定比较。结果表明(1)萌发有助于提高苦荞中芦丁和槲皮素的含量,有助于提高甜荞中芦丁的含量。苦荞萌发7 d后芦丁含量为19.33 mg/g远高于甜荞的0.66 mg/g。(2)苦荞萌发过程中芦丁含量呈先上升后下降再上升的趋势,萌发7 d后芦丁含量提高了66.64%。甜荞萌发7 d后,芦丁含量提高了75.13%。苦荞萌发7 d,槲皮素含量整体呈上升趋势,由8.93 mg/g上升至11.90 mg/g。甜荞萌发过程中未检测出槲皮素。荞麦萌动后提高了黄酮类成分,苦荞中的芦丁含量萌发前后均高于甜荞,选择苦荞作为荞麦萌动食品原料具有更好的营养价值。     

荞麦麸皮提取物对α- 葡萄糖苷酶活性的影响

本实验分别用乙醇和水从荞麦麸皮中提取到了以荞麦黄酮为主要成分的乙醇粗提物和以荞麦糖醇为主要成分的水粗提物,并将乙醇粗提物经高压水解,获得乙醇粗提物的水解物。研究这3 种提取物对α- 葡萄糖苷酶活性的影响,并以阿卡波糖和D- 手性肌醇为参照。比色法和液相色谱法测得,苦荞麸皮乙醇粗提物的主要成分为苦荞黄酮,黄酮含量为74.0%,其中86.5% 为芦丁,槲皮素和异槲皮素微量;乙醇粗提物经高压水解后,总黄酮含量为76.2%,其中,芦丁、槲皮素和异槲皮素分别占60.6%、25.2%、13.5%。甜荞麸皮水粗提物的主要成分为D- 手性肌醇、肌醇,其含量分别为43.6%、2.9%。3 种荞麦麸皮提取物均具有抑制α- 葡萄糖苷酶的活性,作用强度分别为：乙醇粗提物的水解物＞乙醇粗提物＞水粗提物, IC50 分别为0.0085、0.0578、17.3551g/L。苦荞黄酮经高压水解后,抑制α- 葡萄糖苷酶活性显著提高,与阿卡波糖IC50(0.0335g/L)相比,约是它的1/4;水粗提物在低质量浓度下抑制效果不明显,在较高的质量浓度下则表现出抑制活性。     

不同熟化方式对苦荞粉品质的影响

以苦荞粉为原料,研究炒制、烘烤、蒸煮、挤压膨化4种不同熟化方式对苦荞粉的基本营养成分、氨基酸、芦丁、槲皮素、抗氧化活性、水溶性(WSI)、吸水性(WAI)以及色泽的影响.结果 表明,熟化方式会不同程度地降低基本营养成分、氨基酸、芦丁、槲皮素含量等.其中烘烤方式中芦丁、槲皮素含量分别为9.39,0.18 mg/g,与未熟...     

雪松松针乙醇提取物的抗氧化活性及构效关系研究

研究雪松松针乙醇提取物的抗氧化活性及构效关系。通过体外化学分析法,测定雪松松针乙醇提取物的抗氧化活性;采用福林酚法和三氯化铝法,测定其总酚和总黄酮含量;利用高效液相色谱-质谱联用(HPLC-MS)与核磁共振(NMR)技术,鉴定其主要成分,并对其抗氧化作用的构效关系进行初步探讨。结果表明,雪松松针乙醇提取物对实验体系中的ABTS(EC₅₀:0.48±0.01 mg/mL)和DPPH(EC₅₀:32.40±0.76 μg/mL)自由基有很好的清除作用,能够高效地清除羟自由基(EC₅₀:1.73±0.01 mg/mL)和过氧化氢(EC₅₀:89.48±0.54 μg/mL)中的活性氧,抑制脂质过氧化(EC₅₀:9.85±0.14 mg/mL)并具有较高的还原能力,展现出显著的抗氧化活性;提取物拥有较高的总酚(40.30±2.13 mg没食子酸当量/g提取物)和总黄酮(56.62±1.53 mg芦丁当量/g提取物)含量;甲基松柏苷、阿魏酸-β-D-吡喃葡萄糖苷、荛花酚、断氧化马钱苷、(E)-1-O-对香豆酰-α-D-吡喃葡萄糖苷和天竺葵素-3-O-葡萄糖苷是松针乙醇提取物的主要成分,这些化合物结构中大量存在的具有氢原子转移或单电子转移能力的酚羟基、甲氧基、苄基等基团,是其展现良好抗氧化活性的重要原因。     

天山茶茎黄酮苷元提取工艺优化及其生物活性

为了优化天山茶藨茎5种黄酮苷元（木犀草素、槲皮素、山奈酚、异鼠李素、芹菜素）的酸水解提取工艺,评价其体外抗氧化和α-葡萄糖苷酶抑制活性,本文利用UPLC-MS/MS法测定天山茶藨茎5种黄酮苷元的提取率,通过单因素实验和正交试验优化天山茶藨茎黄酮苷元酸水解提取工艺。利用DPPH、ABTS自由基清除实验测定天山茶藨茎提取物及5种黄酮苷元的抗氧化活性,并测定了其α-葡萄糖苷酶抑制活性。结果表明,5种黄酮苷元的检出限、定量限分别在0.8~1.6、2.9~5.4 μg/L,在3.2~207.0 μg/L范围内具有良好的线性关系（R2≥ 0.9962）,方法的稳定性、精密度和重复性良好（RSD≤4.4%）。确定最佳酸水解提取工艺为:90%甲醇溶液（含5%盐酸）、提取温度:70℃、提取时间:100 min,该条件下,总黄酮苷元提取率为4.06 mg/g,RSD<5%,表明正交试验优化的提取条件稳定可行。天山茶藨茎提取物及5种黄酮苷元均具有一定的α-葡萄糖苷酶抑制活性和抗氧化活性,其中甲醇盐酸提取物与5种黄酮苷元的α-葡萄糖苷酶抑制活性均高于阳性对照阿卡波糖（IC₅₀:53.84±2.41 mg/L）,槲皮素对DPPH自由基的清除活性最好（IC₅₀:（2.94±0.18） mg/L）,木犀草素、山奈酚对ABTS自由基清除活性较好（IC₅₀分别为（5.34±0.10）、（5.55±0.17） mg/L）,它们的抗氧化活性高于阳性对照V_C。本研究建立的UPLC-MS/MS方法灵敏、精确、高效,可以对天山茶藨茎5种黄酮苷元化合物同时进行定量分析。优化的酸水解提取工艺能有效提高天山茶藨茎黄酮苷元的提取率。     

Reduction of rutin loss in buckwheat noodles and their physicochemical characterisation

Tartary buckwheat was subjected to hydrothermal treatments for minimising rutin loss in buckwheat-based foods by water addition. When native buckwheat flour was mixed with water for 60 min, the rutin content was distinctly reduced from 3.74 g/100 g to 0.31 g/100 g, increasing the amount of quercetin. However, the rutin content remained constant and quercetin was hardly detected in hydrothermally-treated buckwheat flour. Also, when noodles were prepared with wheat and buckwheat flours (7:3, w/w), the noodle samples containing hydrothermally-treated buckwheat flour, showed higher amounts of rutin (more than 0.83 g/100 g) than the control noodle with native buckwheat flour (0.27 g/100 g). In addition, the use of hydrothermally-treated buckwheat flour gave less pasting parameters and lower viscoelastic properties. The noodle dough with hydrothermally-treated buckwheat flour also had greater water absorption and development time during mixing while the elongation stress of the noodle dough was reduced.     

In vitro potential of flavonoids from tartary buckwheat on antioxidants activity and starch digestibility

The effects of various flavonoids of Tartary buckwheat (rutin-enhanced flavonoid extract [REFE and quercetin-enhanced flavonoid extract [QEFE]) and individual flavonoids (rutin, quercetin and kaempferol) on the antioxidant activity, inhibition of α-glucosidases and α-amylase and starch digestibility were evaluated. Quercetin possessed the highest antioxidant activity and inhibition of α-glucosidases and α-amylase activity followed by kaempferol and rutin. REFE and QEFE have similar antioxidant and inhibition of α-amylase activities, but QEFE has much higher α-glucosidases inhibition than REFE. Tartary buckwheat flour has the lowest content of rapidly digestible starch and predicted glycaemic index (pGI) compared to maize flour, wheat flour and rice flour. Addition of rutin and quercetin to wheat flour showed a weak or no effect on digestion inhibition, but they inhibited starch digestion under solid complex conditions. Our results may help explain the benefits of supplementing the diet with food rich in flavonoids.     

糖苷结构对苦荞黄酮促进葡萄糖在C2C12小鼠骨骼肌细胞中摄取作用的影响

苦荞是芦丁含量最高的粮食作物,但不同的加工过程会导致芦丁降解为单糖苷结构的异槲皮素及其前体槲皮素,进而影响生物活性。本研究比较芦丁及其降解产物（槲皮素、异槲皮素）这三类含有不同糖苷结构的黄酮在C2C12小鼠骨骼肌细胞中促进葡萄糖摄取的功效差异,并探究其作用机制。研究结果表明,苦荞与水接触会导致苦荞中的核心黄酮组分-芦丁降解为槲皮素。芦丁及其降解产物（槲皮素、异槲皮素）均能有效促进C2C12细胞对葡萄糖的摄取,作用顺序为:槲皮素>异槲皮素>芦丁。糖苷键的增加会降低槲皮素促进骨骼肌细胞糖摄取的效果。芦丁和异槲皮素不能激活胰岛素依赖型信号通路中的IRS-1表达及AKT的磷酸化,但高浓度下（400 μmol/L）,芦丁和异槲皮素可以通过非胰岛素依赖型的信号通路中的p-AMPK/p-ACC促进葡萄糖摄取作用,且异槲皮素的作用大于芦丁。槲皮素虽然抑制了IRS-1的表达及AKT的磷酸化,但槲皮素通过AMPK/ACC的磷酸化显著了促进C2C12细胞对葡萄糖的摄取。本研究揭示芦丁及其降解产物在细胞水平上的降血糖作用及机理的差异,对于充分理解苦荞黄酮降血糖机制提供了科学依据。     

Composition and antioxidative properties of the flavonoid-rich fractions from tartary buckwheat grains

Tartary buckwheat, a healthy food, is associated with a reduced risk of certain human chronic diseases; however, its bioactive components are unknown. This study used high performance liquid chromatographyphotodiode array-mass spectroscopy (HPLC-PDA-MS) to identify the flavonoids in flavonoid-rich fractions (FRFs) from tartary buckwheat grain and tested antioxidative capacity of FRFs to confirm their bioactivity. Ultraviolet (UV) absorption and electrospray ionization (ESI⁻) MS/MS spectra identified several flavonols in the embryo, endosperm, testa, and hull, including the predominant flavonoid rutin and minor flavonoids quercetin 3-O-rutinoside-3′-O-β-glucopyranoside, kaempferol 3-O-rutinoside, and quercetin. Tartary buckwheat FRFs extended the Rancimat induction period of lard less than common buckwheat FRF did, while their scavenging rate of DPPH free radicals exceeded that of common buckwheat FRF and rutin. These results indicate that tartary buckwheat FRFs from the grains have potential health benefits.     

加工方式对苦荞中黄酮类化合物的影响研究进展

苦荞是传统的食药两用资源,含丰富的营养与功能成分,具有降糖、降脂、降压和抗炎等多种活性,已被开发成各类食品。黄酮类化合物是苦荞中主要的生物活性物质,对人体健康具有促进作用。芦丁和槲皮素是苦荞黄酮类化合物最主要的存在形式,不同加工方式对其含量及转化有显著影响,从而影响苦荞及其相关制品的功能活性和口感;然而目前缺乏相关的系统论述,不利于差异化苦荞健康产品的开发。本文综述了苦荞中黄酮类成分的稳定性和生物转化特性,以及制粉、萌发、发酵、热处理等几种常见加工方式对苦荞中黄酮类成分的影响规律,对苦荞加工过程中黄酮类成分的保持与转化具有重要指导作用,为开发针对性强的苦荞健康制品提供参考。     

苦荞类黄酮的研究现状及展望

苦荞麦(Tartary buckwheat)为蓼科(polygonaceous)荞麦属双子叶植物,是我国西南地区重要的集营养与保健为一体的粮食作物。现代医学研究表明,苦荞具有抗氧化、降血糖、降血脂、抗肿瘤等多种生理功能,而发挥这些生理功能的物质主要是苦荞中所含有的芦丁、槲皮素等类黄酮化合物。近年来,随着我国人民生活水平的提高,慢性疾病的发病率呈快速增长势头,苦荞以其特有的生理功能逐渐受到人们的青睐。本文综述了苦荞类黄酮的化学组成、生理功能以及加工方式对苦荞制品类黄酮的影响,提出了苦荞类黄酮的研究与开发策略。     

超声辅助苦荞中蛋白和黄酮的同步提取工艺

苦荞中不仅蛋白质所占比例较大,含有丰富的必需氨基酸,也含有其他谷物中没有的芦丁、槲皮素等黄酮类物质,具有较高营养价值和保健功能,可以预防多种慢性疾病。本研究以优化超声辅助进行苦荞中荞麦蛋白和黄酮类化合物的同步提取工艺为目的。根据粉碎粒度、液料比、pH、时间、温度五个单因素试验,对粉碎粒度、pH、温度进行三因素三水平的响应面分析。结果表明苦荞中荞麦蛋白和黄酮类化合物的最佳同步提取工艺条件为:粉碎粒度60目,液料比30∶1mL/g,pH 9.15,提取时间40min,超声温度67.2℃。在此条件下苦荞蛋白得率验证值为7.79%,苦荞黄酮得率验证值为1.14%。可通过该研究得出超声辅助苦荞中蛋白和黄酮的最佳同步提取工艺条件。     

Identification of tartary buckwheat tea aroma compounds with gas chromatography-mass spectrometry

Tartary buckwheat tea, which is an important and healthy product, has a distinct malty aroma. However, its characteristic aroma compounds have not been elucidated. The aims of present study were identification and quantification of its aroma compounds. The analyses were performed by gas chromatography-mass spectrometry (GC-MS) after 3 different isolation techniques. Seventy-seven compounds were identified. Among these compounds, 35 were quantified by available standards. The compounds with a high probability of contribution to the tartary buckwheat tea aroma (OAV ≥ 10) were as follows: 2,5-dimethyl-4-hydroxy-3(2H)-furanone, nonanal, 2,3-diethyl-5-methylpyrazine, benzeneacetaldehyde, maltol, 2,5-dimethylpyrazine, 2-ethyl-5-methylpyrazine, trimethylpyrazine. Some nutritional and bioactive compounds were also identified in this study, such as linoleic acid, niacin, vanillic acid, 7-hydroxycoumarin, butylated hydroxytoluene. Practical Application: Tartary buckwheat, one type of buckwheat, has gained much attention from nutritionists and medical doctors in recent years. It is rich in rutin, quercetin, and other nutrients that are good for health. Tartary buckwheat-based product such as tartary buckwheat tea is an important and popular healthy product in China, Japan,South Korea, European countries as well as in American countries. It has a distinct malty aroma. The present study first identified and quantified of its aroma compounds. The results will draw attention to other researchers in food flavor and buckwheat filed.