响应面优化千日红酪氨酸酶抑制活性成分的提取工艺

本文研究了千日红粗提物对酪氨酸酶的抑制作用并采用响应面法优化千日红中酪氨酸酶抑制剂的提取工艺。在单因素实验的基础上,探讨了提取温度、提取时间、液料比和乙醇浓度等因素为影响因子,应用Box-Benhnken中心组合法进行4因素3水平试验设计,以酪氨酸酶抑制率率为响应值,进行响应面分析,优化千日红中酪氨酸酶活性抑制组分的提取工艺。结果表明,最佳提取条件:提取温度为80℃、提取时间为3 h、液料比为60 mL/g(V/M)、乙醇浓度为50%(V/V)。在此条件下,RSM模型预测千日红乙醇粗提物对酪氨酸酶的抑制率为54.94%,实测千日红乙醇粗提物对酪氨酸酶的抑制率为(53.86±2.11)%,与预测值无显著性差异(p0.05)。     

白附子酪氨酸酶活性抑制成分的提取与分离

研究了不同溶剂及提取方式对白附子中酪氨酸酶抑制成分的影响,并应用活性追踪法对白附子中酪氨酸酶活性抑制成分进行考察。结果表明,以20%乙醇水溶液为溶剂从白附子中提取的粗提物对酪氨酸酶活性的抑制作用最强,抑制率可达到96.35%,与恒温水浴振荡提取法相比,超声波辅助提取法对活性成分的提取效果更好。在此提取条件下所得白附子粗提物对酪氨酸酶的半抑制浓度IC50为24.69mg/mL。该粗提物依次经乙醚和正丁醇萃取,所得正丁醇萃取组分对酪氨酸酶的抑制率达71.94%。对高活性的正丁醇萃取组分进行分离,获得了一种酪氨酸酶抑制率高达98.82%的组分,是一种潜在的酪氨酸酶抑制剂。研究结果对白附子及其活性组分在医药和食品领域的应用有指导意义。     

菠萝酪氨酸酶抑制活性组分的研究

以酶抑制动力学为跟踪方法,考察了菠萝中具双孢菇酪氨酸酶抑制作用的活性成分,并将之用于双孢菇的抗褐变,结果显示对酪氨酸酶有抑制作用的活性成分存在于乙醇组分中,且该活性成分对双孢菇酪氨酸酶的抑制作用属于不可逆抑制;5%菠萝汁对双苞菇浆褐变的抑制率54.8%,0.4%乙醇组分65.1%.     

4种茶叶醇提物对酪氨酸酶的抑制作用

采用提取分离法提取土豆中的酪氨酸酶,并以多巴为底物,运用酶活性测定法研究4种茶叶醇提物对土豆酪氨酸酶活性的抑制作用.结果表明,在4种茶叶的50%乙醇(V/V)提取液中,生药质量浓度达到4.0 mg/mL时,茶叶醇提物对土豆酪氨酸酶的活性有不同程度的抑制作用,其中青茶对酪氨酸酶的抑制作用最大,抑制率为62.28%.     

孜然乙醇提取物的工艺优化及对酪氨酸酶活性的抑制作用

以孜然为原料,在单因素实验的基础上,以酪氨酸酶抑制率为指标,采用响应面法优化孜然乙醇提取物的提取工艺,并测定不同极性部位对酪氨酸酶的影响及孜然乙醇提取物中主要活性成分的含量。结果表明,孜然乙醇提取物抑制酪氨酸酶的最优提取工艺为:乙醇体积分数71%,液料比16:1 (mL/g),提取时间2.2 h,提取温度83 ℃,在此条件下其对酪氨酸酶活性的抑制率达(42.61%±0.56%),与预测值43.20%接近,表明响应面分析的结果可靠;孜然乙醇提取物中多酚、黄酮、皂苷、多糖四种活性成分的含量分别为(1.35%±0.03%)、(4.40%±0.29%)、(0.76%±0.04%)、(4.89%±0.28%);孜然乙醇提取物不同极性部位对酪氨酸酶均有抑制作用,且呈浓度依赖关系,其IC₅₀大小顺序为:水相 > 石油醚相 > 正丁醇相 > 乙酸乙酯相。因此,优化得到的孜然乙醇提取物抑制酪氨酸酶的提取工艺合理、可行,乙酸乙酯和正丁醇部位对酪氨酸酶活性的抑制能力较强,多酚、黄酮、皂苷、多糖等都可能是孜然乙醇提取物抑制酪氨酸酶的功效成分。     

胡芦巴不同溶剂粗提物对两种储粮害虫的驱避和触杀活性

为开发环境和谐的植物性杀虫剂,以乙醇、乙酸乙酯、丙酮、氯仿、石油醚为溶剂采用温浸回流法对胡芦巴Trigonella foenum-graecum L.种子中的生物活性物质进行提取;并以主要储粮害虫杂拟谷盗Tribolium confusum du Val、谷蠹 Rhyzopertha dominica Fbricius成虫为试验对象,采用滤纸药膜法对胡芦巴5种溶剂粗提物进行驱避、触杀活性测定.结果表明:5种溶剂粗提物对杂拟谷盗的驱避作用均优于对谷蠹的作用:在各粗提物剂量为1.6 μg/cm2时,对杂拟谷盗处理7 d后的平均驱避率均超过80%,驱避等级为Ⅴ级,其中丙酮粗提物效果最好.但5种溶剂粗提物对谷蠹的触杀作用却均优于对杂拟谷盗的作用:处理72 h后,乙醇、乙酸乙酯、丙酮、氯仿粗提物对谷蠹成虫的LD50依次为0.112、0.102、0.106、0.100 mg/cm2,石油醚粗提物的触杀活性相对较差;乙醇、乙酸乙酯、氯仿粗提物用0.91 mg/cm2剂量处理72 h后,杂拟谷盗成虫的平均死亡率分别为95.56%、72.22%、87.78%;丙酮、石油醚粗提物用1.04 mg/cm2剂量处理72 h后,杂拟谷盗成虫的平均死亡率分别为57.78%和53.33%.     

油茶籽壳粗提物的抗氧化活性研究

采用不同浓度的乙醇溶液提取油茶籽壳中的有效成分,通过自由基体系、双氧水体系和亚硝酸盐体系研究油茶籽壳粗提物的抗氧化性能.结果表明,提取物的抗氧化活性与乙醇浓度有关,以60%乙醇提取物的抗氧化活性最强,当浓度为15、2、4、10 g/L时,粗提物对·OH、DPPH·、H2O2和亚硝酸盐的清除率最高分别为47.7%、76%、97.1%和99.0%,且粗提物用量与清除能力呈现明显的量效关系.     

甘薯叶提取物对酪氨酸酶的抑制作用

为探究甘薯叶提取物对酪氨酸酶活性的影响,利用不同极性溶剂萃取甘薯叶乙醇提取物,分别得到石油醚层、三氯甲烷层、乙酸乙酯层及正丁醇层萃取物,并采用多巴速率氧化法考察不同溶剂提取物对酪氨酸酶的抑制作用。结果表明,甘薯叶提取物中存在能抑制酪氨酸酶活性的物质,其中甘薯叶正丁醇萃取物对酪氨酸酶的抑制作用最明显,根据正丁醇萃取物抑制酪氨酸酶的Lineweaver-Burk双倒数拟合方程可得,甘薯叶提取物对酪氨酸酶的抑制作用是可逆过程,其抑制类型是混合型,KI与KIS分别为0.198、0.209 mg/mL,即甘薯叶提取物与游离酶的结合程度大于与酶-底物络合物的结合。     

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。     

藜麦皂苷的提取及其酪氨酸酶抑制活性

探究藜麦"陇藜1号"籽粒中皂苷的最佳提取条件并验证其酪氨酸酶抑制活性。设计三因素三水平正交实验,探索料液比,乙醇浓度及超声时间对藜麦皂苷提取得率的影响。构建L-多巴诱导的B16细胞模型,测定藜麦皂苷对细胞活性及酪氨酸酶活性的影响,最终通过免疫印迹法(Westernblot)探索藜麦皂苷影响酪氨酸酶活性的作用通路。结果显示:藜麦皂苷最佳提取条件为料液比1:40,70%浓度的乙醇下超声60 min,得率为17.85 mg/g。当藜麦皂苷的浓度低于200μg/m L时,其对B16细胞无显著抑制作用,但能显著抑制酪氨酸酶的活性和黑色素的形成,200μg/m L藜麦皂苷处理B16细胞72 h,黑色素相对含量降至73.85%,酪氨酸酶活性降至53.31%。藜麦皂苷通过抑制小眼畸型相关转录因子(MITF)及酪氨酸酶(TYR)蛋白表达来抑制黑色素形成。藜麦皂苷可以作为一种具有美白活性的组分在化妆品或药品中应用。     

体外消化对藜蒿叶多酚含量及抗氧化活性的影响

本文研究了藜蒿叶提取物在模拟体外消化过程中酚酸含量、抗氧化活性以及对酪氨酸酶活力抑制率的变化。研究表明,经过模拟体外消化后藜蒿叶提取物中的绿原酸、1,5-双咖啡酰奎宁酸、3,4-双咖啡酰奎宁酸、3,5-双咖啡酰奎宁酸、4,5-双咖啡酰奎宁酸5种酚酸含量均显著性降低。总酚含量和DPPH、FRAP、ABTS抗氧化活性经过模拟体外消化后均显著性增加。消化后粗提物和纯化样总酚含量分别升高了73.57%、23.79%,DPPH抗氧化能力分别增强了36.17%、53.29%,FRAP抗氧化能力分别增强了28.71%、15.45%,ABTS清除自由基能力分别增强了163.67%和49.50%。此外,藜蒿叶粗提物对酪氨酸酶活力的抑制率在体外消化过程中也显著性增加,消化后粗提物的抑制率可达到49.88%。从而可有效抑制黑色素生成,具有较好的美白潜力。因此,藜蒿叶提取物经体外消化后具有较好的抗氧化活性和抑制酪氨酸酶活力的能力,有望应用于美白产品的开发,该研究为藜蒿茎加工副产物-藜蒿叶的开发提供了借鉴。     

赤芍抗氧化活性及其成分研究(英文)

利用DPPH法、β-胡萝卜素法和还原力法评估赤芍提取物的抗氧化活性。结果显示乙酸乙酯提取物具有最高的抗氧化活性。利用硅胶,RP-18和Sephadex LH-20对其进行活性追踪分离,得到四种纯的抗氧化活性成分。利用1D、2DNMR和ESI-MS对其结构进行鉴定。结果显示为没食子酸、儿茶精、没食子酸芍药甙和没食子酸氧化芍药甙。其抗氧化活性均大于同浓度的人工合成抗氧化剂BHT。实验证明赤芍及其提取物是一种高效价廉的天然的抗氧化剂,可用作食品添加剂。     

ANTIOXIDANT ACTIVITY OF PHENOLIC FRACTIONS OF WHITE BEAN (PHASEOLUS VULGARIS)

An extract from white bean seeds was prepared using 80% (v/v) acetone. Four fractions (I-IV) -were separated from the crude extract on a Sephadex LH-20 column using methanol as the mobile phase. The antioxidant activity of fractions was investigated in a β-carotene-linoleate model system. For individual fractions, IV spectra were recorded and the content of total phenolics was determined. Fractions were also characterized based on the number of phenolic compounds, and their antioxidant activity determined by TLC analysis. The presence of caffeic, p-coumaric, ferulic, and sinapic acids in the form of free and estrified compounds was found in fraction IV. One dominant phenolic compound was present in fraction III after acid hydrolysis with a maximum absorption at 278 nm. Results of the β-carotene-linoleate model system indicated that antioxidant activity of separated fractions did not correlate exactly with their content of total phenolic compounds and were in the order of IV>III>II>I. Individual fractions contained several phnolic compounds as noted by TLC. Spots on the plates sprayed with a solution of p carotene-linoleate indicated that these compounds can act as natural antioxidants. Absorption maxima in the W spectra showed that jlavonoids, and not phenolic acids, were the main phenolic compounds present in the separated fractions.     

Isolation and identification of two novel umami and umami-enhancing peptides from peanut hydrolysate by consecutive chromatography and MALDI-TOF/TOF MS

Peanut hydrolysate produced by crude protease extract from Aspergillus oryzae HN 3.042 was found to elicit intense umami and umami-enhancing effect. Taste profiles, amino acid and organic acid composition of peanut hydrolysate and its separation fractions by ultrafiltration were evaluated. The results revealed that peanut hydrolysate was mainly low molecular weight compounds. Fractions of 1-3 kDa and below 1 kDa prominently contributed to the umami taste and umami-enhancing effect of the peanut hydrolysate. The two fractions were further purified, using gel filtration chromatography and reverse-phase high-performance liquid chromatography (RP-HPLC), in combination with sensory evaluation, to obtain a umami peptide and umami-enhancing peptide. The active peptides were identified as Ser-Ser-Arg-Asn-Glu-Gln-Ser-Arg (SSRNEQSR, 963.9 Da) and Glu-Gly-Ser-Glu-Ala-Pro-Asp-Gly-Ser-Ser-Arg (EGSEAPDGSSR, 1091.1 Da), by MALDI-TOF/TOF MS, respectively.     

ORAChromatography and total phenolics content of peanut root extracts

A large number of compounds have been reported in peanut plants. Many of these compounds are phytoalexins, which are produced by plants experiencing environmental stress and often exhibit antioxidant activity. It is difficult to determine which of the many compounds has the greatest impact on total antioxidant capacity in a mixture. The objectives of this research were to examine the oxygen-radical absorbing capacity (ORAC) value and total phenolic contents of peanut root extracts and peanut root extract fractions collected via HPLC. Peanut roots were extracted from four different cultivars (Brantley, NC-12, Phillips, and Wilson) with 70% aqueous ethanol with ultrasonic assistance. Each cultivar was sampled in duplicate. The extracts were fractionated into 18 3-min fractions by HPLC using a C-18 column. Fractions and crude extracts were freeze dried. ORAC values and total phenolic content were then determined for all fractions and crude extracts. Fractions had a significant effect on the μM TE/mg gallic acid equivalents (GAE). ORAC values ranged from -46.89 μM TE to 185 μM TE in HPLC fractions. ORAChromatography can be used to focus on antioxidants in complex samples.     

Antioxidant activities of phenolic rich fractions (PRFs) obtained from black mahlab (Monechma ciliatum) and white mahlab (Prunus mahaleb) seedcakes

The antioxidant activities of phenolic rich fractions (PRFs) from crude methanolic extract (CME), and its fractions using ethyl acetate (EAF), hexane (HF) and water (WF) of black mahlab (Monechma ciliatum) and white mahlab (Prunus mahaleb) seedcakes were investigated. The total phenolic compounds were found to be higher in white mahlab than black mahlab seedcakes. The antioxidant activity determined by the DPPH method revealed that black mahlab PRFs had the highest antioxidant activity, compared to white mahlab fractions. The presence of antioxidants in the two mahlab PRFs reduced the oxidation of β-carotene by hydroperoxides from these extracts/fractions. The effect of the two mahlab PRFs on the oxidative stability of corn oil at 70 °C was tested in the dark and compared with butylated hydroxyanisole (BHA). The CME performed better antioxidant activity in inhibiting the formation of both primary and secondary oxidation products. The qualitative and quantitative characterisation of phenolic compounds was carried out by HPLC/DAD.     

Biological activity of rice extract and the inhibition potential of rice extract,rice volatile compounds and their combination against α-glucosidase, α-amylase and tyrosinase

Rice is produced for consumption and traditional medicine. Rice is also used as an ingredient in cosmetic products. In this study, the author investigated the biological activity and inhibition potential against α-glucosidase, α-amylase and tyrosinase activity of rice extract (black rice [BR], red rice [RR] and white rice [WR]), rice volatile compounds, rice extract combined with volatile compounds, rice extract combined with standard inhibitors and volatile compounds combined with standard inhibitors. The results revealed that the free-radical scavenging capacity of rice extract is related to the phenolic content and flavonoids. BR showed the highest potential to inhibit α-glucosidase and α-amylase activity, whereas WR showed the highest potential to inhibit tyrosinase activity. Among rice volatile compounds, vanillin and vanillyl alcohol had the highest inhibition potential against α-glucosidase and α-amylase, respectively, whereas guaiacol had the highest inhibitory activity against tyrosinase. Molecular docking supported by the high binding efficiency was also obtained from vanillin and guaiacol when located at the active site of these enzymes. The combination of RR with acarbose (AB) had the highest inhibition potential and showed a synergic effect on both α-glucosidase and α-amylase. Interestingly, the combination of rice extract (BR, RR and WR) and vanillin and vanillyl alcohol had a synergic effect on α-amylase. Moreover, the combination of WR and vanillyl alcohol had the highest inhibition potential and showed a synergic effect on tyrosinase, whereas rice volatile compounds had a synergic effect on tyrosinase obtained from 2-pentylfuran/kojic acid (KA), vanillin/KA and vanillyl alcohol/KA.     

北京早园竹叶不同提取组分对CHO细胞Akt信号通路的影响

通过比较北京早园竹叶提取物中的主要活性组分黄酮苷类、酚酸类及总提取物3个组分对中国仓鼠卵巢细胞(CHO)的增殖、凋亡及Akt信号通路作用差别,考察竹叶提取物对正常细胞可能产生有害作用的活性组分。采用MTT法检测细胞增殖作用、Annexin V-FITC/PI双染流式法测细胞凋亡、In Cell Analyzer 1000活细胞图像分析系统测定Akt信号通路的活化。结果显示：1)50～400μg/mL范围内3种组分对细胞存活率的影响无明显变化,当质量浓度≥800μg/mL时细胞增殖率明显下降(P＜0.05),细胞毒性作用明显,且以酚酸类组分作用最强;2)不同组分的竹叶提取物在200～800μg/mL范围内与空白组比较均可降低细胞凋亡率(P＜0.05),但总提取物和黄酮苷类在质量浓度为1600μg/mL时本身可诱导CHO细胞的凋亡增加,差异显著(P＜0.05),且以黄酮苷类诱导的细胞凋亡率最高,达14.21%;3)与对照组比较,黄酮苷类在100～800μg/mL剂量条件下,对Akt的活性无明显影响,总提取物在400、800μg/mL时Akt的激活率明显增加(P＜0.01),而酚酸类在100～800μg/mL范围内,随剂量增加,激活率增高,呈明显的剂量-反应关系(r=0.996,P＜0.01)。由此可见3种不同提取物组分对CHO细胞的作用是有差别的,其中酚酸类可能是产生有害作用的主要组分。