笃斯越桔花青素在不同pH条件下热稳定性的研究

研究分析了笃斯越桔花青素提取物(BBAE)在不同pH条件下热稳定性的变化,测量指标包括吸光值、颜色及其ABTS自由基清除能力.研究结果表明:在pH4～6加热时,BBAE的吸光值无明显变化,提示其在酸性条件下有较好的稳定性.在pH7和pH8加热时,尤其在pH7时,BBAE的吸光值明显降低,表明BBAE在pH7条件下的热稳定性最差.通过色差仪测定BBAE溶液的L*、a*、b*值从而算出H(色调)值.随着加热温度的升高,pH为4～8时的BBAE溶液的H值呈现不同程度的升高,且逐渐趋近于90°.这表明在加热过程中,BBAE溶液的颜色由最初的红色逐渐向黄色转变.pH为7时的BBAE溶液在吸光值降低,颜色变黄的同时,其ABTS自由基清除率也有显著性的下降.     

越橘提取物中花青素分析及其体外抗氧化活性

研究越橘提取物中花青素的组成与含量及其体外抗氧化活性。实验采用高效液相色谱与质谱联用法对越橘提取物中的花青素组成进行鉴定,并以矢车菊素-3-O-葡萄糖苷为标准品测定了花青素的含量。以V_C为对照,测定越橘提取物对DPPH自由基、ABTS自由基、羟基自由基的清除能力和还原力。结果表明,越橘提取物中含有14种花青素,包括飞燕草素、矢车菊素、矮牵牛素、芍药素和锦葵素类;越橘提取物中花青素的总含量为408.74 μg/mg,其中,矢车菊素类花青素含量最高,为159.93 μg/mg,占总含量的39.12%;体外抗氧化实验表明,越橘提取物的DPPH自由基清除能力、ABTS自由基清除能力和羟基自由基清除能力的IC₅₀分别为26.16、13.07、1.91 mg/mL,还原力在浓度为250 μg/mL时达到0.617。本研究为越橘的开发利用提供一定理论与数据基础。     

Bog Bilberry (Vaccinium uliginosum L.) Extract Reduces Cultured Hep-G2, Caco-2, and 3T3-L1 Cell Viability,Affects Cell Cycle Progression,and Has Variable Effects on Membrane Permeability

ABSTRACT: Bog bilberry (Vaccinium uliginosum L.) is a blue-pigmented edible berry related to bilberry (Vaccinium myrtillus L.) and the common blueberry (Vaccinium corymbosum). The objective of this study was to investigate the effect of a bog bilberry anthocyanin extract (BBAE) on cell growth, membrane permeability, and cell cycle of 2 malignant cancer cell lines, Caco-2 and Hep-G2, and a nonmalignant murine 3T3-L1 cell line. BBAE contained 3 identified anthocyanins. The most abundant anthocyanin was cyanidin-3-glucoside (140.9 ± 2.6 μg/mg of dry weight), followed by malvidin-3-glucoside (10.3 ± 0.3 μg/mg) and malvidin-3-galactoside (8.1 ± 0.4 μg/mg). Hep-G2 LC50 was calculated to be 0.563 ± 0.04 mg/mL, Caco-2 LC50 was 0.390 ± 0.30 mg/mL and 0.214 ± 0.02 mg/mL for 3T3-L1 cells. LDH release, a marker of membrane permeability, was significantly increased in Hep-G2 cells and Caco-2 cells after 48 and 72 h compared to 24 h. The increase was 21% at 48 h and 57% at 72 h in Caco-2 cells and 66% and 139% in Hep-G2 cells compared to 24 h. However, 3T3-L1 cells showed an unexpected significant lower LDH activity (P ≤ 0.05) after 72 h of exposure corresponding to a 21% reduction in LDH release. BBAE treatment increased sub-G1 in all 3 cell lines without influencing cells in the G2/M phase. BBAE treatment reduced the growth and increased the accumulation of sub-G1 cells in 2 malignant and 1 nonmalignant cell line; however, the effect on membrane permeability differs considerably between the malignant and nonmalignant cells and may in part be due to differences in cellular membrane composition.     

pH值对乳清蛋白糖基化产物体外抗氧化特性的影响

选择6种不同的糖基配体,通过湿热糖基化反应,研究乳清蛋白不同糖基化复合物在不同pH值下的反应特性及抗氧化特性。结果表明:在pH值3～9的条件下,6种乳清蛋白糖基化复合物与乳清蛋白420nm和294nm处的吸光值差异显著(P<0.05);且6种乳清蛋白糖基化复合物的游离氧基酸含量均显著降低(P<0.05)。除了乳清蛋白-蔗糖复合物外,其他5种乳清蛋白糖基化复合物的还原能力、DPPH自由基清除能、·OH自由基清除能力和抗脂质过氧化能力随pH值的升高都有所提高。乳清蛋白-木糖复合物的抗氧化能力提高最大,而且在pH值7～9的反应条件下,具有比V_c更高的还原能力。     

‘蓓蕾’蓝靛果中花色苷组成鉴定及抗氧化能力比较分析

对蓝靛果中花色苷的组成进行鉴定,并对其抗氧化能力进行比较分析。实验以蓝靛果（‘蓓蕾’品种）为原料,采用有机溶剂60%乙醇（0.1%盐酸酸化）溶液,超声辅助提取90 min;利用D101大孔树脂对获得的粗提物进行纯化,之后冷冻干燥制得粉末物质。通过pH示差法和福林-酚法分别测定总花色苷含量和总多酚含量,分别为（353.35±0.79）、（474.01±2.12）mg/g;并用高效液相色谱-质谱联用法对花色苷组成进行鉴定,共发现11 种花色苷,其中矢车菊-3-葡萄糖苷为主要花色苷（90.679%）。此外,实验还通过总抗氧化能力测定和2,2’-联氨-二（3-乙基苯并噻唑-6-磺酸）二铵盐自由基、1,1-二苯基-2-三硝基苯肼自由基清除能力测定,比较分析蓝靛果花色苷提取物、矢车菊-3-葡萄糖苷、VC的抗氧化能力,结果表明,3 种物质的抗氧化能力排序为：矢车菊-3-葡萄糖苷＞花色苷提取物＞VC。     

桑椹提取物中酚类化合物的抗氧化及抗糖基化活性分析

利用葡聚糖凝胶层析柱将桑椹提取物分为6 个组分（F1、F2、F3、F4、F5、F6）,研究了桑椹中酚类化合物的抗氧化能力和抗糖基化能力。采用Folin-酚法、pH示差法和高效液相色谱法测定各组分总酚、总花色苷含量和酚类化合物成分;通过1,1-二苯基-2-三硝基苯肼（1,1-diphenyl-2-picrylhydrazyl,DPPH）自由基、2,2’-联氮-二（3-苯并噻唑-6-磺酸）（2,2’-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid),ABTS）自由基清除率和亚铁还原能力实验评价各组分抗氧化能力;建立牛血清白蛋白-果糖模拟反应体系和牛血清白蛋白-丙酮醛模拟反应体系评价各组分抗糖基化能力。结果表明：桑椹提取物中检测出5 种酚酸、两种花色苷和两种黄酮类物质;F3的总花色苷含量最高（P＜0.05,（101.10±2.39） mg 矢车菊素-3-葡萄糖苷当量/g）,F5的总酚含量最高（P＜0.05,（188.05±2.01） mg 没食子酸当量/g）;F3的抗氧化能力显著高于其他组分（P＜0.05,DPPH自由基清除能力、ABTS＋·清除能力、亚铁还原能力分别为（129.33±5.58）、（194.33±2.48）、（130.44±1.38） mmol Trolox/g）,F5的抗糖基化能力显著强于其他组分（P＜0.05,牛血清白蛋白-果糖模拟反应体系和牛血清白蛋白-丙酮醛模拟反应体系中的糖基化抑制率分别为（87.23±0.36）%和（66.99±1.62）%）;桑椹提取物的抗氧化能力、抗糖基化能力分别与总花色苷、总酚含量呈现显著的回归关系（P＜0.05）。     

Anthocyanin characterization and bioactivity assessment of a dark blue grained wheat (Triticum aestivum L. cv. Hedong Wumai) extract

Defatted bran of dark blue grained wheat (Triticum aestivum L., cv. Hedong Wumai) was extracted using ethanol and chemically characterized by HPLC and ESI-MS. Cyanidin-3-glucoside was the predominant anthocyanin in the pigmented wheat which also contained cyanidin-3-galactoside, pelargornidin-3-glucoside, and peonidin-3-glucoside. Ferulic acid was identified as the major simple phenolic acid, with lesser amounts of p-hydroxybenzoic acid, caffeic acid, syringic acid and p-coumaric acid also present. Anthocyanins were separated from phenolic acids using solid phase extraction, and respective fractions were determined for DPPH radical, ABTS radical scavenging tests as well as oxygen radical absorption assay. Results showed that 69% of the overall free radical scavenging capacity of dark blue grained wheat was attributed to the anthocyanin content, compared to 19% for the extractable phenolic acids. In cell based models, the pigmented wheat extract significantly suppressed both hydrogen peroxide-induced intracellular oxidation (p < 0.01) and bacterial lipopolysaccharide-induced nitric oxide (p < 0.05) production. These data report for the first time the major phytochemical composition of pigmented grained wheat and associated bioactivity towards free radical scavenging and suppression of ROS and RNS activity.     

蔓越莓花色苷的组成鉴定及抗氧化能力

对蔓越莓花色苷的组成进行鉴定,并对其抗氧化能力进行测定。采用pH示差法测定花色苷提取量,超高压辅助提取蔓越莓花色苷含量为（75.49±0.43）mg/100?g,常规溶剂提取蔓越莓花色苷含量为（67.31±1.08）mg/100?g,蔓越莓中总花色苷含量为（79.52±0.50）mg/100?g;选择AB-8大孔树脂对蔓越莓花色苷粗提物进行纯化,冻干粉中花色苷含量从（46.10±0.92）mg/g提高到（309.26±2.37）mg/g。通过测定1,1-二苯基-2-三硝基苯肼自由基清除率、2,2’-联氮基-双-（3-乙基苯并噻唑啉-6-磺酸）二铵盐自由基清除能力和总抗氧化能力,比较蔓越莓花色苷与VC的抗氧化能力。结果表明：同质量浓度条件下,蔓越莓花色苷的抗氧化能力强于VC。通过高效液相色谱-质谱联用技术在蔓越莓中鉴定出7?种花色苷：芍药素-3,5-二己糖苷、矢车菊素-3-半乳糖苷、矢车菊素-3-葡萄糖苷、矢车菊素-3-阿拉伯糖苷、芍药素-3-半乳糖苷、芍药素-3-葡萄糖苷、芍药素-3-阿拉伯糖苷,其中芍药素-3,5-二己糖苷首次在蔓越莓中被鉴定出。     

酰基稳态化花色苷的制备及其热降解特性和抗氧化活性

食品级的肉桂酸甲酯为酰基供体,运用减压酶法,催化其与矢车菊素-3-O-葡萄糖苷（C3G）发生酰化反应,具有绿色安全、高效且酰化产物单一等优点。对酰化产物进行分离,并进行热稳定性和抗氧化性等性质的测定。结果表明：酰化转化率为80%;利用半制备高效液相色谱仪对酰化后的产物进行分离纯化,纯化后的酰化C3G纯度达98.3%;经质谱分析可知,酰化后产物为矢车菊素-3-(6-肉桂酰)-葡萄糖苷（C3(6C)G）。酰基化修饰使花色苷的脂溶性、热稳定性得到了明显改善。采用1,1-二苯基-2-三硝基苯肼（1,1-diphenyl-2-picrylhydrazyl,DPPH）自由基清除、2,2-氨基-二(3-乙基-苯并噻唑啉-6-磺酸)（2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid,ABTS）阳离子自由基清除、铁离子还原/抗氧化能力法（ferric reducing and antioxidant power,FRAP）、羟自由基清除体外实验测定不同浓度下C3G、C3(6C)G和VC的抗氧化能力。与相同浓度的VC相比,除了在ABTS阳离子自由基清...     

Saskatoon berries (Amelanchier alnifolia Nutt.) scavenge free radicals and inhibit intracellular oxidation

Two cultivars (Thiessen and Smoky) of Saskatoon berries (Amelanchier alnifolia Nutt.) possess free radical scavenging activities in a concentration-dependent manner, whereas Thiessen presented higher (p < 0.05) activity compared to Smoky due to its relatively abundant anthocyanin content. Total anthocyanin content significantly corresponded to free radical scavenging activities (p < 0.05). Solvent fractionation revealed that the free radical scavenging components, i.e., anthocyanin, predominantly occurred in the ethyl acetate and n-butanol extracted fractions, suggesting that active components are more likely to occur in glycoside forms. HPLC further confirmed the existence of cyanidin-3-O-galactoside and cyanidin-3-O-glucoside as leading anthocyanins in the Saskatoon berries. Furthermore, Saskatoon berries extracts from both cultivars suppressed peroxy-radical induced intracellular oxidation in a concentration-dependent manner without affecting cell viability. The obtained results provided useful information for the future application of Saskatoon berries in the nutraceutical industry.     

黑米与欧洲越橘花色苷抗氧化活性比较研究

为探究黑米与欧洲越橘花色苷抗氧化活性的差异,以抗坏血酸为阳性对照,通过比较其总还原力及清除超氧阴离子、羟自由基、过氧化氢、DPPH·、ABTS+·的能力,对其体外抗氧化作用进行了比较研究。结果表明,在测试浓度范围内,黑米花色苷与欧洲越橘花色苷总还原能力、清除过氧化氢、DPPH·、ABTS+·能力均高于阳性对照,且黑米花色苷具有更好地清除DPPH·、ABTS+·能力(IC₅₀分别为0.88、0.75 μg/mL);两种花色苷清除羟自由基、超氧阴离子自由基能力均低于阳性对照,但黑米花色苷清除超氧阴离子能力是欧洲越橘的3.63倍,清除羟自由基能力与欧洲越橘花色苷相当。研究表明,黑米花色苷具有更好的抗氧化性,是良好的天然抗氧化剂。     

笃斯越桔花青素提取物对3T3-L1前脂肪细胞生长抑制作用

目的：研究笃斯越橘花青素提取物对3T3-L1 前脂肪细胞生长抑制作用及其作用机理。方法：采用MTT法观察细胞生长抑制并确定半数抑制浓度(IC50),LDH(lactate dehydrogenase)法评价提取物引起的细胞膜损伤,流式细胞计法测定细胞周期,荧光显微镜观察凋亡细胞的形态学特征。结果：笃斯越橘花青素提取物能够有效地抑制3T3-L1 前脂肪细胞的生长且IC50 约为214μg/mL;LDH 实验表明细胞LDH 释放率在24、48、72h 分别为89.0%、85.2%、67.8%,72h 时相对24h 或48h 明显降低(P ＜ 0.05),提取物对3T3-L1 前脂肪细胞膜的通透性未见明显增加;笃斯越橘花青素提取物能够有效诱导细胞凋亡,同时在S 或G0/G1 期产生阻滞,但是对细胞的G2/M 期未见明显影响,荧光显微镜观察细胞出现典型凋亡特征。结论：笃斯越橘花青素提取物能有效抑制3T3-L1 前脂肪细胞生长的作用,具有开发为天然抗肥胖功能因子的潜在可能。     

改性乳清蛋白体外抗氧化特性

以木糖、葡萄糖、果糖、乳糖、麦芽糖、蔗糖和乳清蛋白为美拉德反应原料,按照一定比例进行湿热糖基化反应。结果表明:在湿热糖基化反应后,6种糖类与乳清蛋白复合物在420 nm和294 nm处的吸光值差异显著(P<0.05);除乳清蛋白-蔗糖复合物外,其他5种乳清蛋白糖基复合物的游离氨基酸含量均显著降低(P<0.05)。除了乳清蛋白-蔗糖复合物外,其他5种乳清蛋白糖基化复合物随浓度的增加,还原能力和DPPH自由基清除能力都有所提高,其中乳清蛋白-木糖复合物的抗氧化能力提高最大,而且SDS-PAGE显示乳清蛋白-木糖复合物反应后分子质量明显增大;傅里叶红外光谱(FTIR)结果显示,乳清蛋白-木糖和乳清蛋白-葡萄糖复合物中蛋白质的酰胺I和II明显减少。     

The effects of six phenolic acids and tannic acid on colour stability and the anthocyanin content of mulberry juice during refrigerated storage

Six phenolic acids and tannic acid are added to fresh mulberry juice to enhance the colour. Effects of these copigments on colour stability and the anthocyanin content of mulberry juice are investigated while being stored at 4 °C for 30 days. Results suggest that the addition of the copigments markedly increases the maximum absorbance of mulberry juice. Cyanidin-3-glucoside is the major anthocyanin, followed by cyanidin-3-rutinoside. Furthermore, the retention rate of the total anthocyanin, cyanidin-3-rutinoside and cyanidin-3-rutinoside increases after the addition of all copigments. The effect of ferulic acid on the stability of total anthocyanin, cyanidin-3-glucoside and cyanidin-3-rutinoside in mulberry juice is the best, 89.52%, 86.90% and 94.46% respectively (66.56%, 62.58% and 73.60% in the control group). Researchers recommend the addition of ferulic acid, p-coumaric acid and caffeic acid – all of which belong to the hydroxycinnamic acid group, to the mulberry juice to enhance the colour stability and anthocyanin content.     

黑果腺肋花楸花色苷提取工艺优化及其抗氧化活性和组成鉴定

目的：优化超声波辅助提取黑果腺肋花楸花色苷的条件,测定花色苷的抗氧化能力,鉴定黑果腺肋花楸花色苷提取物的组成成分。方法：采用响应面法优化花色苷提取条件,通过1,1-二苯基-2-三硝基苯肼自由基清除能力、2,2’-联氨-双（3-乙基苯并噻唑啉-6-磺酸）二铵盐自由基清除能力及Fe3＋还原能力方法评价其抗氧化能力,高效液相色谱-质谱联用法进行成分分析。结果表明：提取温度60?℃、超声功率100?W、料液比1∶30（g/mL）、超声时间31?min、乙醇体积分数64%和pH?2.0时为最优提取条件,此时黑果腺肋花楸花色苷含量可达（3.61±0.01）mg/g。体外抗氧化实验表明,在相同质量浓度条件下,黑果腺肋花楸花色苷提取物的抗氧化活性明显高于VC。组分鉴定表明黑果腺肋花楸花色苷提取物中共有6?种花色苷,其中矢车菊-己糖苷二聚体和矢车菊-3,5-二己糖苷为新检测出的2?种花色苷。     

Phenolic Analyses of Haskap Berries (Lonicera caerulea L.): Spectrophotometry Versus High Performance Liquid Chromatography

Haskap berries (Lonicera caerulea L.) are known for their high phenolics, anthocyanins, and antioxidant potential. The data on the phenolic profile of these fruits are lacking. In this study, the phenolic profiles of three haskap varieties; tundra, berry blue, and indigo gem grown in Nova Scotia, Canada were investigated for the first time using spectrophotometery and high-performance liquid chromatography. Berries were analyzed for total phenolic content/total reducing capacity, total anthocyanin content, and antioxidant potential (2,2-diphenyl-1-picrylhydrazyl radical scavenging activity). The total reducing capacity, total anthocyanin content, and 2,2-diphenyl-1-picrylhydrazyl values were 6.17–8.42 mg gallic acid equivalents/gram fresh weight, 4.49–6.97 mg cyanidin-3-glucoside equivalents/gram fresh weight, and 78.70–89.55%, respectively. The extracts were analyzed by reversed-phase diode array detector–high-performance liquid chromatography through a gradient elution using Synergi 4 µm Max-RP C12 column and the chromatograms were acquired at 520, 360, and 320 nm for athocyanins, flavonoids, and free phenolic acids, respectively. The identified anthocyanins were cyanidin-3-glucoside (82.81–91.99% of the total anthocyanins), cyanidin 3,5-di-glucoside (2.31–4.27%), cyanidin-3-rutinoside (1.54–9.20%), peonidin-3-O-glucoside (0.75–3.44%), and pelargonidin-3-glucoside (0.77–2.98%). Other flavonoids (quercetin-3-β-D-glucoside and quercetin-3-rutinoside) and free phenolic acids (chlorogenic and neochlorogenic) were also quantified.     

Antioxidant activity of corozo (<Emphasis Type="Italic">Bactris guineensis</Emphasis>) fruit by electron paramagnetic resonance (EPR) spectroscopy

The antioxidant activity of an anthocyanin-rich extract (ARE) of corozo fruit (Bactris guineensis) and its major constituents, cyanidin-3-rutinoside, cyanidin 3-glucoside and peonidin-3-rutinoside, was measured by stabilised free-radical EPR spectroscopy. The ARE of corozo exhibited a higher radical scavenging activity than pure anthocyanins using ABTS and DPPH free radicals. The antioxidant activity values for pure anthocyanins against the two radicals were different; however, a second-order kinetic model was followed in all of the cases. A cooperative effect regarding free-radical scavenging activity among studied anthocyanins was suggested. This study is the starting point to consider the ARE of corozo fruit as promising antioxidant material to be used in food industry.     

野生蓝果忍冬多酚鉴定及其抗氧化、降血糖活性

目的：对野生蓝果忍冬进行多酚成分鉴定和抗氧化、抗淀粉酶活性研究。方法：以我国小兴安岭地区的野生蓝果忍冬果实为材料,通过固相萃取技术分离纯化得到花色苷和非花色苷分离液,测定分离液中的化学成分,并测定多酚粗提液中总酚和总花色苷含量,抗氧化活性（1,1-二苯基-2-三硝基苯肼自由基清除能力、铁离子还原能力、2,2’-联氮-双(3-乙基苯并噻唑啉-6-磺酸)阳离子自由基清除能力、氧自由基吸收能力）,抗α-淀粉酶、α-葡萄糖苷酶活性和脂肪酶活性。结果：野生蓝果忍冬果实中含有丰富的花色苷和非花色苷多酚,尤其是矢车菊素-3-葡萄糖苷和山柰酚-3-芸香糖苷含量很高;蓝果忍冬多酚粗提液中总酚含量为82.7 mg/100 g,总花色苷含量为49.8 mg/100 g,具有较强的抗氧化活性;对α-淀粉酶、α-葡萄糖苷酶活性的半抑制质量浓度分别为0.39 mg/mL和0.933 mg/mL,对脂肪酶活性的半抑制质量浓度为12.31 mg/mL,说明蓝果忍冬可以作为一种有前景的淀粉酶抑制剂。     

发芽糙米蛋白-葡聚糖共价接枝复合物的开发及其物理化学性能、乳液稳定性的评价

将发芽糙米蛋白（germinated brown rice protein,GBRP）和葡聚糖通过美拉德反应开发一种新型的乳化剂。使用傅里叶变换红外光谱仪和X射线衍射仪分析发芽糙米蛋白-葡聚糖共价复合物（germinated brown rice protein-dextran conjugate,GBRP-Dex Con）。结果表明,通过与葡聚糖共价结合,相比于GBRP,GBRP-Dex Con的溶解度提高了10%（pH 7.0）;变性温度（122.2 ℃）提高了14.7 ℃（GBRP为107.5 ℃）;GBRP-Dex Con清除DPPH自由基能力及总还原能力显著提高（P＜0.05）。通过测定美藤果油乳状液的粒径及物理稳定性发现,GBRPDex Con在稳定乳状液比GBRP、发芽糙米蛋白-葡聚糖混合物（GBRP-Dex Mix）以及作为常规乳化剂的大豆分离蛋白更为有效。本方法制备的乳化剂GBRP-Dex Con具有优异的性能和应用到食品工业的潜力。     

Anthocyanin Pigment Composition of Blackberries

ABSTRACT: The anthocyanin pigments in 51 blackberry samples were isolated and characterized by highperformance liquid chromatography (HPLC) with UV-visible detection, electrospray mass spectroscopy (ESMS), and hydrolysis and saponification reactions. Five pigments were detected: cyanidin-3-glucoside, cyanidin-3-rutinoside, cyanidin-3-xyloside, cyanidin-3-glucoside acylated with malonic acid, and an unidentified acylated derivative of cyanidin-3-glucoside. Four different quantitative patterns were evident among the cultivars and selections. Total anthocyanin content ranged from 70.3 to 201 mg/100 g with a mean of 137 mg/100 g. Ten commercial blackberry juice concentrate samples were analyzed. Total anthocyanin ranged from 12.3 to 107 mg/100 g on a single-strength juice basis, and 1 sample had an atypical profile.