黑米花色苷矢车菊素-3-O-β-葡萄糖苷对脂多糖诱导的人THP-1单核细胞炎症损伤的保护作用

单核细胞介导的炎症反应在动脉粥样硬化发生和发展过程中起关键作用。花色苷是一种具有多种生物学活性的多酚类黄酮化合物,富含于各种深色的蔬菜、水果及谷类中,其中矢车菊素-3-O-β-葡萄糖苷（cyanidin-3-O-β-glucoside,Cy-3-g）是花色苷中重要的单体,本研究旨在探讨黑米来源的Cy-3-g对脂多糖（lipopolysaccharide,LPS）诱导的人单核白血病细胞（Tohoku hospital pediatrics-1,THP-1）炎症损伤的作用及可能的分子机制。采用乙醇-盐酸溶液浸提、大孔树脂吸附洗脱等步骤得到黑米花色苷粗提物,然后用中压液相层析联合紫外检测提取得到纯化的Cy-3-g（纯度>96.5%）。用不同质量浓度（0、0.10、0.25μg/mL和0.50μg/mL）Cy-3-g与THP-1细胞共孵育4 h,然后加入LPS（质量浓度50 ng/mL）与细胞继续孵育48 h,用酶联免疫吸附试验法测定培养液中白细胞介素-1β(interleukin-1β,IL-1β)、IL-6、IL-8、IL-10和肿瘤坏死因子-α(tumor necrosis fac...     

矢车菊素-3-葡萄糖苷通过AMPK抑制成熟脂肪细胞脂蛋白脂酶活性

通过矢车菊素-3-葡萄糖苷(Cy-3-g)干预成熟脂肪细胞,研究Cy-3-g对细胞脂蛋白脂酶(LPL)活性的影响及其作用机制。结果表明:Cy-3-g通过激活一磷酸腺苷激活的蛋白激酶(AMPK)抑制成熟脂肪细胞的LPL活性,提示Cy-3-g具有潜在的调节机体脂代谢作用,与含Cy-3-g的花色苷提取物的肥胖抑制作用密切相关。     

矢车菊素-3-葡萄糖苷对骨骼肌细胞脂蛋白脂肪酶活性的影响

富含矢车菊素-3-葡萄糖苷(Cy-3-g)的花色苷提取物可抑制肥胖,但其作用机制不明。骨骼肌细胞脂蛋白脂肪酶(LPL)活性与肥胖相关,LPL使骨骼肌中的β-脂肪酸氧化增强,从而加速清除体内循环的甘油三酯(TG),抑制TG流向脂肪组织积聚而导致肥胖。本实验建立稳定的骨骼肌细胞分离培养方法,通过花色苷Cy-3-g干预骨骼肌细胞,研究Cy-3-g对细胞LPL活性的影响及其作用机制。结果表明：Cy-3-g通过激活一磷酸腺苷激活的蛋白激酶(AMPK)上调骨骼肌细胞LPL活性,提示Cy-3-g具有潜在的调节机体脂代谢作用,与含Cy-3-g的花色苷提取物的肥胖抑制作用密切相关。     

HPLC法测定黑米矢车菊素-3-葡萄糖苷

建立水浴震荡提取-高效液相色谱法测定黑米中矢车菊素-3-葡萄糖苷(Cy-3-G)的方法。确定了HPLC法测定Cy-3-G的测定条件,通过响应面优化试验确定了水浴震荡提取黑米中Cy-3-G的最优提取条件。实验得HPLC法测定Cy-3-G的条件为:色谱柱(C18,150mm×4.6mm,5um),柱温30℃,进样体积10u L,检测波长520nm,流动相A(甲醇)-B(1%甲酸水溶液),流速1m L/min,梯度洗脱:0~3min,A∶B(20∶80);3~17min,A∶B(20∶80~30∶70);17~20min,A∶B(30∶70~20∶80)。最优提取条件为:溶剂是含0.8%盐酸的50%乙醇水溶液、料液比1∶20、提取次数2次、温度70℃、时间20min,Cy-3-G的得率为0.728%。结果表明水浴震荡提取法实现了黑米中Cy-3-G的高效提取,满足样品测定的前处理需求;建立的HPLC法测定黑米中Cy-3-G的方法操作简便、重复性好、测定结果精密度高。     

桑椹花青素在小鼠消化道中的动力学变化研究

研究了经口摄入桑椹花青素后,桑椹花青素在消化道中的动力学变化.给小鼠灌胃不同剂量的桑椹花青素混合物,采用比色法检测不同时间消化道中残留的花青素总量.并用HPLC法比较灌胃前样品和小鼠粪便中回收的花青素样品中花青素含量的变化.当灌胃剂量低于12.5mg/鼠时,小鼠粪便中残留的花青素保持较低的水平;当灌胃剂量高于12.5mg/鼠时,小鼠粪便中花青素残留量与荆量呈直线相关.前4h,消化道中花青素残留量呈递减趋势,4 h后花青素残留量变化很小.桑椹花青素经过小鼠消化道后,矢车菊素-3-葡萄糖苷和矢车菊素-3-芸香糖苷的比例发生了倒置,说明矢车菊素-3-葡萄糖苷的吸收率远高于矢车菊素-3-芸香糖苷的.     

黑米种皮中花色苷组成及含量的HPLC分析

花色苷是黑米种皮中重要的活性成分,具有多种生物活性,准确分析其组成及含量对黑米的加工利用研究有重要意义。采用酸性甲醇溶液浸提黑米种皮得到粗提液,用高效液相色谱法(HPLC)通过与标准品对照,分析黑米皮中主要花色苷的组成种类及含量,旨在建立一种简便、准确的黑米种皮花色苷分析方法。研究结果表明,黑米种皮中主要含有3种花色苷组份,分别是矢车菊-3-葡萄糖苷(23.30 mg/g),矢车菊-3-芸香糖苷(0.97 mg/g),芍药定-3-葡萄糖苷(2.24 mg/g)。该方法样品前处理简单,是一种简便、准确的花色苷组成及含量的测定方法。     

HPLC法测定不同产地黑豆皮中矢车菊素-3-O-葡萄糖苷的含量

目的：建立黑豆皮中矢车菊素-3-O-葡萄糖苷的含量测定方法,测定5个不同产地黑豆皮中矢车菊素-3-O-葡萄糖苷的含量。方法：采用HPLC法测定黑豆皮中矢车菊素-3-O-葡萄糖苷的含量,色谱柱采用Phenomenex Luna Su C18 柱 (250mm×4.60mm,5μm);流动相A相为0.5%磷酸溶液,B相为水-乙腈(50:50),进行梯度洗脱;流速0.8mL/min;检测波长520nm;柱温30℃;进样量10μL。结果：矢车菊素-3-O-葡萄糖苷标准曲线回归方程为：Y＝2×107X－33120(r＝0.9998),在 0.1041～1.041μg范围内线性关系良好,平均回收率分别为92.4%、 92.5%和95.5%,不同产地黑豆皮中矢车菊素-3-O-葡萄糖苷含量范围为5.263～12.829mg/g。结论：所用方法简便、准确,可用于不同产地黑豆皮的质量控制。     

有色大米中花青素UPLC测定方法的建立

从样品处理和色谱分析两个方面进行优化,建立了有色大米中花青素超高效液相色谱(UPLC)分析方法。样品经处理后,在5 min内即可完成色谱分析,添加回收率高于95%;日内和日间相对标准偏差分别为2.97%和5.87%; 6种花青素的最低检出限介于0.08～0.15 mg/kg。运用该方法进行检测表明:黑米中花青素总量最高,为3 251.90 mg/kg;其次为紫米,而红米中花青素含量较低;黑米和紫米中花青素种类主要为矢车菊素和芍药素;不同产地的紫米中花青素总量差异明显,且紫米中花青素主要分布在外层。     

超高效液相色谱法测定有色稻米中花色苷的含量

采用盐酸甲醇溶液超声提取有色稻米花色苷,以超高效液相色谱-紫外检测器对有色稻米中主要花色苷矢车菊素-3-O-葡萄糖苷和芍药素-3-O-葡萄糖苷进行定量检测。结果表明,在0. 5～50. 0μg/mL浓度范围内线性关系良好,R~2 0. 999;矢车菊素-3-O-葡萄糖苷回收率在93. 0%～98. 5%,RSD在0. 33%～3. 50%;芍药素-3-O-葡萄糖苷回收率在96. 0%～111. 7%,RSD在0. 42%～2. 18%。对四川重庆地区的4个黑米样品、1个紫米样品和5个红米样品的糙米和米糠中主要花色苷的测定结果表明,黑米、黑米糠、紫米和紫米糠中花色苷总量分别为0～1 051. 27 mg/kg,74. 06～206. 82 mg/kg,296. 18 mg/kg和116. 15 mg/kg;红米中未检出矢车菊素-3-O-葡萄糖苷和芍药素-3-O-葡萄糖苷,部分红米糠中检出矢车菊素-3-O-葡萄糖苷,含量为0～50. 94 mg/kg。     

聚酰胺树脂分离纯化黑米中矢车菊素-3-葡萄糖苷工艺研究

通过单因素试验和正交试验,确定了聚酰胺树脂分离纯化黑米中矢车菊素-3-葡萄糖苷的最佳工艺参数。以矢车菊素-3-葡萄糖苷得率为考察指标时,其最佳工艺条件为:上样液pH4、上样液质量浓度25mg/ml、上样液流速6ml/min、洗脱液乙醇体积分数50%、洗脱流速3ml/min;以纯度为考察指标时,该最佳工艺条件为:上样液pH3、上样液质量浓度25mg/ml、上样液流速3ml/min、洗脱液乙醇体积分数30%、洗脱流速6ml/min。聚酰胺树脂能有效纯化黑米花色苷,最终所得样品纯度达到80.84%,大部分黄酮类杂质被有效去除。     

Hepatoprotective and antioxidant activity of anthocyanins in black rice bran on carbon tetrachloride-induced liver injury in mice

The benefits of anthocyanin-rich black rice bran extract (ARBE) on the liver of carbon tetrachloride (CCl₄)-intoxicated mice was investigated. Based on the in vivo experiment, the effect of cyanidin-3-glucoside (Cy-3-G) and peonidin-3-glucoside (Pn-3-G), the predominant anthocyanins in ARBE, on CCl₄ insulted hepatocytes L-02 was further evaluated. Mice treated with ARBE for 7 weeks by oral administration showed reduced aminotransferase activities in serum, accompanied by enhanced superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activity, while tiobarbituric acid reactive substances (TBARS), and 8-hydroxy-2′-deoxyguanosine (8-OHdG) levels were significantly decreased compared to CCl₄-intoxicated model group. Histopathological observation showed ARBE administration alleviated the pathological changes in livers of CCl₄-intoxicated mice. Similarly, preincubation of L-02 cells with Cy-3-G or Pn-3-G significantly alleviated CCl₄-induced injury dose-dependently, exhibited higher cell viability, decreased aminotransferase activity and enhanced cellular antioxidant status. Furthermore, Cy-3-G showed much stronger hepatoprotective activity than Pn-3-G at the same concentration. HPLC analysis showed that Cy-3-G accounted for more than 88% of total anthocyanins in ARBE. These results indicate that ARBE is significantly beneficial to liver health, and that Cy-3-G is the predominant anthocyanin in ARBE exerting this effect. The antioxidant activity of anthocyanins is an important mechanism by which ARBE exerts hepatic health benefit.     

黑米花色苷的提取及纯化

目的：得到黑米花色苷最佳提取工艺,建立应用大孔吸附树脂纯化花色苷的方法。方法：以矢车菊素-3- 葡萄糖苷为跟踪指标,通过单因素试验和正交试验,对影响黑米花色苷提取的各因素进行研究,比较9 种大孔吸附树脂对花色苷的静态吸附和解吸性能。结果：黑米花色苷最佳提取条件,提取液乙醇- 水- 盐酸体积比为50:50:0.5,温度50℃,固液比为1:10(g/mL),提取时间为1h,提取次数为3 次。通过对9 种大孔吸附树脂的比较,确定AB-8 为纯化黑米的理想吸附树脂,80% 乙醇为洗脱剂,上样流速为1.0BV/h,解吸流速为2.0BV/h。结论：测定经树脂纯化后提取物中花色苷的含量达到22.59%(粗提物中花色苷含量为3.448%),树脂富集倍数为 6.02,此工艺条件纯化效果显著。     

HPLC法测定黑米皮提取物中花色苷成分及含量

目的:用高效液相色谱法测定黑米皮提取物中花色苷的成分及含量。方法:采用HypersilBDSC18柱(250×4.6mm,5μm),以乙腈-4%磷酸:(15:85)为流动相,流速0.8ml/min,检测波长530nm,用外标法定量。结果与结论:黑米皮提取物中主要含两种花色苷:矢车菊定-3-葡萄糖苷和芍药定-3-葡萄糖苷,其含量分别为25.7%和1.7%。     

Comparative studies on major nutritional components of black waxy rice with giant embryos and its rice bran

The concentration of nutrients in brown rice is mainly associated with embryo size. Various beneficial components have been purified from rice bran. Recently developed black waxy rice with a giant embryo (‘Milyang 263’, BGE), which is the ge ^t mutant of the GE gene, was selected and analyzed to produce high quality nutritional components. γ-Aminobutyric acid (GABA) content in BGE rice bran, one of the most important nutritional compounds in rice, showed a 10.6-fold increase (2.66±0.48 mg/g) compared to that of BGE brown rice (0.25±0.01 mg/g). In addition, brown rice and BGE rice bran showed enriched amylopectin (94.5±0.5 and 97.0±0.0%) and bioactive anthocyanin [Cy-3-G: 75.15±4.18 (brown rice), 82.97±0.81 (rice bran) and Pn-3-G: 1.52±0.50 (brown rice), 4.33±0.20 (rice bran); mg/100 g] contents. These results suggest that BGE rice bran can be used as an excellent raw material to efficiently produce high quality essential amino acids, cyanidin-3 glycoside-enriched anthocyanins, and GABA.     

Compositions of anthocyanins in blackberry juice and their thermal degradation in relation to antioxidant activity

This study investigated the compositions of anthocyanins in blackberry juice and the effect of heating (70, 80, and 90?°C) on individual anthocyanins, as well as its impact on antioxidant activity. The results showed that five anthocyanins were identified, including cyanidin (Cy)-3-O-glucoside (1), Cy-3-O-arabinoside (2), Cy-3-O-malonyl-glucoside (3), Cy-3-O-dioxalyl-glucoside (4), and the Cy derivative (5). The degradation rates of individual anthocyanins (1–4) followed the first-order reaction kinetics. Unexpectedly, the amount of the Cy derivative (5) was observed to slightly increase in temperature. At the same temperature, the half-lives (T _1/2) values of each anthocyanin (1–4) significantly differed (P?<?0.05). Maximum of the T _1/2 values were observed with Cy-3-O-dioxalyl-glucoside (T _1/2 values ranged 9.48–3.54?h) and exhibited relatively resistant stability. Moreover, the T _1/2 values of Cy-3-O-glucoside were higher than Cy-3-O-arabinoside and Cy-3-O-malonyl-glucoside. The antioxidant activity of blackberry juice reduced with the decreasing of total anthocyanins. These results demonstrated that the time dependency of anthocyanins degradation and antioxidant capacity were strongly correlated in blackberry juice.     

‘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。     

In Vitro Antioxidant Activity of the Anthocyanins in Sageretia theezans Brongn Fruit

Antioxidant activities of the extract containing anthocyanins from fruit of Sageretia theezans Brongn were investigated in this work. The fruit of Sageretia theezans Brongn was treated with 0.001 mol·L^–1 HCl in a solid (g): liquid (mL) ratio 1:10 to give extract. Total anthocyanins content was 253.10 ± 2.31 mg in 100 g fruit. The anthocyanins in extract were determined as cyanidin-3-sophorose-5-glucoside (Cy-3-Sp-5-Glu), petunidin-3-(6′-malonyl)-glucoside (Pt-3-(6’-Mal)-Glu), malvinidin-3-glucosid (Mv-3-Glu), and peonidin-3-(6′-malonyl)-glucoside (Pn-3-(6’-Mal)Glu) in 0.38, 22.57, 44.32, and 30.86%, respectively, by high performance liquid chromatography-mass spectrometry method. Antioxidant activities of the anthocyanins in extract were evaluated by methods of ferric reducing antioxidant power, hydroxyl radical scavenging assay, and superoxide radical scavenging assay. The anthocyanins in extract exhibited obvious antioxidative activities. These results suggested that the fruit of Sageretia theezans Brongn could be considered as a good source of natural antioxidants.     

Anti-oxidative analysis,and identification and quantification of anthocyanin pigments in different coloured rice

Anthocyanin pigments in coloured rice cultivars were isolated and identified using high-performance liquid chromatography techniques. Two black rice cultivars (Asamurasaki, Okunomurasaki) contained three major anthocyanins: cyanidin-3-glucoside, peonidin-3-glucoside and malvidin. Chinakuromai (black) rice additionally contained a fourth anthocyanin, petunidin-3-glucoside. Four red rice cultivars contained only malvidin. The total anthocyanin content varied greatly among black rice cultivars (79.5–473.7 mg/100 g), but was lower in red rice (7.9–34.4 mg/100 g). Total phenolic content was similar between red (460.32–725.69 mg/100 g) and black (417.11–687.24 mg/100 g) rice. The oxygen radical absorbing capacity was ranked as follows: red (69.91–130.32 μmol Trolox/g) > black (55.49–64.85 μmol Trolox/g) > green (35.32 μmol Trolox/g) > white (21.81 μmol Trolox/g) rice. The antioxidant capacity resulted mainly from the seed capsule, not the endosperm. The anthocyanin pigments contributed little to the total antioxidant capacity of red (0.03–0.1%) and black (0.5–2.5%) rice cultivars. Hence, the antioxidant capacity is derived mainly from other phenolic compounds.     

Comparison of the activities of hydrophilic anthocyanins and lipophilic tocols in black rice bran against lipid oxidation

The antioxidant capabilities of anthocyanin and tocol extracts from black rice bran were evaluated using an emulsion system containing either cholesterol (1.0 mg/ml) or fish oil (10 mg/ml). The cholesterol oxidation product, 7-ketocholesterol, increased to 180.1 μg/ml in the control emulsion after 168 h of oxidation, while it was only 15.4 and 39.0 μg/ml in the emulsions containing 1 μg/ml of the anthocyanin and tocol extracts, respectively; but below 1.2 μg/ml in the emulsion having 5 μg/ml of anthocyanins or tocols. In the fish oil emulsion, over 80% of C20:5 and C22:6 were oxidised after a 48 h incubation at 37 °C, while they were retained above 38% and 65% in the emulsions containing 10 μg/ml of anthocyanins and tocols, respectively, and above 85% in the emulsion containing 20 μg/ml of anthocyanins or tocols. Compared with the tocols extract, the capability of the anthocyanin extract was relatively greater in stabilising cholesterol but lower in inhibiting fatty acids oxidation.