Folin-Ciocalteu比色法测定花椒中多酚含量的研究

研究了以没食子酸为标准品,采用Folin-Ciocalteu比色法,测定花椒多酚含量的适宜条件。结果表明:当Folin-Ciocalteu试剂与10%Na2CO3溶液之比为1∶2(V∶V),Folin-Ciocalteu试剂体积为4 mL,反应温度为25℃,反应时间为120 min时,多酚含量在1~6μg/mL范围内与吸光值呈良好的线性关系。测定了4种花椒中的多酚含量,四川青花椒多酚含量最高。     

Folin-Ciocalteu比色法测定桦褐孔菌多酚的条件优化

以没食子酸为标准品,通过正交试验和单因素实验研究了Folin-Ciocalteu 比色法测定桦褐孔菌中多酚含量的适宜条件。结果表明,优化后的显色条件为Folin-Ciocalteu试剂用量0.3mL、10% Na2CO3溶液0.6mL,25℃时避光反应30min,于750nm处测定其吸光值。多酚质量浓度在0.0032－0.0256mg/mL范围内与吸光值有良好的线性关系。根据拟合的线性回归方程对桦褐孔菌多酚进行定量测定,野生桦褐孔菌和人工培养桦褐孔菌中多酚含量最高分别为（72.05±0.08）mg/g、（52.76±0.06）mg/g（n=6）。不同加标水平的加标回收率测定实验获得的平均回收率为98.95%,RSD为1.27%。该法测定桦褐孔菌多酚具有快速、准确,稳定性强,重现性好,精密度高的特点,可应用于实际样品的测定。     

Folin-Ciocalteu分光光度法测定夏枯草总酚含量的研究(英文)

通过采用Folin-Ciocalteu试剂,以没食子酸为对照品,用紫外可见分光光度法,研究测定夏枯草中总酚含量。结果表明,在装有样品的10 mL容量瓶中依次加入Folin-Ciocalteu试剂0.5 mL,20%Na2CO31.7 mL,室温放置60 min后,在波长660 nm或760 nm测定吸光度。多酚质量浓度在0～10.3μg/mL范围内与吸光度有良好的线性关系,回归方程在660 nm为Y=94.542X-0.0067,R2=0.9999,加样回收率为101.7～105.0%;760 nm为Y=101.13X-0.0191,R2=0.9990,加样回收率为105.1～107.6%。本研究在稳定性、准确性和重复性方面都具有较好的实验结果,可为夏枯草多酚的定量分析提供方法参考。     

固液态发酵条件对桑叶发酵提取物中降糖活性物质的影响

目的:探讨不同发酵条件对桑叶总黄酮、总多酚含量及体外降糖活性的影响。方法:本研究主要利用比色法考察固态、液态两种发酵条件对桑叶发酵物中总黄酮、总多酚含量的影响,同时建立α-葡萄糖苷酶和α-淀粉酶为体外降糖活性筛选模型,以期筛选最佳发酵条件。结果:液体发酵7 d桑叶70%乙醇提取物对α-葡萄糖苷酶抑制率最高(92.79%);未发酵水提取物对α-淀粉酶抑制率最高(63.68%),其次为液体发酵7 d桑叶70%乙醇提取物(54.81%)。且在生药浓度为10 mg·m L-1时,都高于阳性对照阿卡波糖。结论:利用冠突散囊菌发酵,可以改变桑叶提取物的体外降糖活性,为开发糖尿病药物提供可行思路。     

桑叶多酚的研究进展

桑叶作为一种药食两用的资源,在我国栽培历史悠久,种植面积较广。植物多酚是一类广泛存在于植物体内的重要次生代谢产物,具有多种生物活性,近年来越来越受到人们关注。本文对桑叶多酚的提取纯化、含量测定、成分鉴定和生物活性的最新研究进展进行了综述,并展望了桑叶多酚研究应用的前景。     

复合酶法提取黄秋葵嫩果多酚的工艺优化

对复合酶法提取黄秋葵嫩果多酚的工艺进行优化。通过单因素及正交试验,确定复合酶中纤维素酶、甘露聚糖酶和果胶酶的最佳配比浓度,分别是2%、3%、1%;在此基础上,以提取温度、料液比、提取时间为自变量,黄秋葵多酚提取含量为响应值,采用Design-Expert软件中的Central Composite Design模块设计3因素组合试验,建立多项式回归方程,经响应面回归分析优化组合条件,得到复合酶法提取黄秋葵嫩果多酚的最佳提取工艺为:提取温度44.97℃,液料比(mL:g)58.80∶1,提取时间3.99 h,预测多酚提取值为29.193 mg/g,验证值为29.156 mg/g。研究结果可为黄秋葵多酚的批量提取工艺提供参考。     

桑叶中聚戊烯醇结构与含量测定

本文建立了高效液相色谱法确定桑叶聚戊烯醇的异戊烯基单元数和含量的方法,采用Thermo C18ODS-2(250 mm×4.6 mm,5μm)色谱柱,流动相为甲醇-异丙醇(8∶7,v/v),流速为1.50 mL/min,柱温为25℃,检测器为PDA。测定结果表明,桑叶中聚戊烯醇的异戊烯基单元数为10~12,聚戊烯醇含量为0.731%。该方法准确度高、重现性好,能快速测定桑叶中聚戊烯醇的异戊烯基单元数和含量,为桑叶资源的开发提供理论依据。     

响应曲面优化醇法提取桑叶黄酮工艺研究

以含水乙醇为溶剂,在单因素试验基础上,采用响应曲面法优化提取桑叶黄酮工艺条件。结果表明,醇法提取桑叶黄酮的最优工艺条件为:乙醇体积分数71.75%、提取温度67.1℃、料液比23.2:1(醇溶液:桑叶粉,mL:g)、提取时间150 min、提取次数2,桑叶黄酮得率为2.37%。验证试验表明,该优化工艺稳定,与模型预测值相符。     

新鲜烟叶中12种多酚类化合物的UPLC-MS-MS分析

多酚类物质含量高低是衡量烟叶质量的重要指标,快速准确测定其含量可为烟叶质量评价和多酚代谢研究提供技术支持.本研究将25 mg新鲜烟叶冻干粉末样品经5 mL 50％甲醇超声20 min提取后离心,上清液经0.22 μm滤膜过滤后用超高效液相色谱-串联质谱仪(UPLC-MS-MS)在多反应监测(MRM)模式下分析.研究发现,咖啡酸、阿魏酸、莨菪亭和香豆素等4种低含量多酚化合物在0.05～5.00 μg/mL范围内线性良好;新绿原酸、莨菪苷、山奈酚、山奈酚苷和山奈酚-3-O-芸香糖苷等5种中等含量多酚化合物在0.2～40.0 μg/mL范围内线性良好;绿原酸、隐绿原酸和芸香苷等3种高含量多酚化合物在5～200 μg/mL范围内线性良好.12种多酚的回收率在84.7％～102％,相对标准偏差(n=6)在1.9％～5.7％,定量限在0.5～8.0μg/g.研究结果表明,本方法具有灵敏度高、专一性好和结果准确等特点,适用于新鲜烟叶中12种多酚含量的高通量分析,可为新鲜烟叶代谢过程中多酚含量的变化研究提供有效分析手段.     

新鲜烟叶中12种多酚类化合物的UPLC-MS-MS分析

多酚类物质含量高低是衡量烟叶质量的重要指标,快速准确测定其含量可为烟叶质量评价和多酚代谢研究提供技术支持.本研究将25 mg新鲜烟叶冻干粉末样品经5 mL 50％甲醇超声20 min提取后离心,上清液经0.22 μm滤膜过滤后用超高效液相色谱-串联质谱仪(UPLC-MS-MS)在多反应监测(MRM)模式下分析.研究发现,咖啡酸、阿魏酸、莨菪亭和香豆素等4种低含量多酚化合物在0.05～5.00 μg/mL范围内线性良好;新绿原酸、莨菪苷、山奈酚、山奈酚苷和山奈酚-3-O-芸香糖苷等5种中等含量多酚化合物在0.2～40.0 μg/mL范围内线性良好;绿原酸、隐绿原酸和芸香苷等3种高含量多酚化合物在5～200 μg/mL范围内线性良好.12种多酚的回收率在84.7％～102％,相对标准偏差(n=6)在1.9％～5.7％,定量限在0.5～8.0μg/g.研究结果表明,本方法具有灵敏度高、专一性好和结果准确等特点,适用于新鲜烟叶中12种多酚含量的高通量分析,可为新鲜烟叶代谢过程中多酚含量的变化研究提供有效分析手段.     

Selective transport of the mulberry leaf urease from the midgut into the larval hemolymph of the silkworm,Bombyx mori

Just before spinning, larvae of the silkworm, Bombyx mori, absorb intact urease of the host plant (mulberry leaf) from the midgut lumen into the hemolymph. In order to investigate whether the transport of the mulberry leaf urease is selective, crude proteins extracted from the mulberry leaves were labeled with biotin and orally administered to the fifth instar larvae. The biotinylated proteins transported into the hemolymph were detected by ligand blotting using streptavidin. When the biotinylated proteins were administered to 5-day-old fifth instar larvae, a strong signal of a biotinylated protein was detected in the hemolymph 2 days after the administration. In contrast, when the biotinylated mulberry leaf proteins were administered to 3-day-old fifth instar larvae, no signal derived from the biotinylated proteins was detected in the hemolymph. The signal weakened when the biotinylated proteins had been immunoprecipitated before administering to the larvae, indicating that the signal came from the mulberry leaf urease. These results show that the transport of the mulberry leaf urease from the midgut into the hemolymph is selective and larval-stage specific. Subsequently, binding assays were carried out to test the binding ability of the mulberry leaf urease to the brush border membrane in the epithelial cells of larval midgut. The urease was not bound to the brush border membrane vesicles (BBMV) from the midgut of 3-day-old fifth instar larvae, while more than 60% of the total amount of incubated urease was bound to the BBMV from the midgut of 6-day-old fifth instar larvae. The urease binding ability of BBMV correlated with the uptake of the mulberry leaf urease. This suggests that a urease binding molecule(s) exists in the BBM of the midgut epithelium, which is involved in the uptake of the mulberry leaf urease. In addition, the uptake of the mulberry leaf urease into the hemolymph was induced by 20-hydroxyecdysone.     

油松不同部位多酚与单宁的含量比较研究

油松为我国特有树种,药用历史悠久且分布广泛,其药用部位含有多酚和单宁等生物活性物质。本文采用Folin-Cicalten法和柠檬酸铁铵法对油松不同部位多酚与单宁含量进行了测定。实验结果表明,油松根、茎和叶多酚含量依次为89.24±12.06、68.18±11.20、104.30±12.51 mg·g^-1;单宁含量依次为25.38±5.85、20.92±3.55、39.02±5.68 mg·g^-1。油松多酚与单宁在油松各部位有相同的分布趋势,均为叶中最高,根次之,茎中最低。与其他中草药植物多酚含量相比,油松多酚含量较高,具有较大的开发前景和应用价值。     

HPLC-ELSD法测定家桑野桑植株中1-脱氧野尻霉素的含量

采用HPLC-ELSD法对桑植株各部位所含的1-脱氧野尻霉素(DNJ)进行了含量测定,使用TSKgel Am-ide-80(4.6 mm×250 mm,5μm)色谱柱,流动相为:乙腈-水(84∶16,6.5 mM醋酸铵),流速为1.00 mL/min,柱温:30℃,线性范围为0.505μg～20.2μg(r=0.9991,n=6),平均回收率为94.95%(n=5),RSD=1.98。测定结果表明,野桑叶、嫩枝和根皮部位的DNJ含量较高,而家桑的叶、枝皮和根皮中含量较高。本方法准确度高、重现性好,能快速检测桑树资源及类似植物和相关产品中1-脱氧野尻霉素的含量。     

桑-蚕系统中镉的吸收、累积与迁移

对桑－蚕系统中镉的吸收,累积与迁移行为研究结果表明：（１）桑树对土壤镉污染有一定的耐性,桑树镉累积量和相对累积率与土壤镉浓度的关系可分别用回归方程Ｔａ＝ａ＋ｂｌｏｇ（Ｓｃ）和ｌｏｇ（Ｒａ）＝ａ＋ｂｌｏｇ（Ｓｃ）描述。（２）随着土壤镉浓度的增加,镉在桑树根部的分布率明显增加,地上部分的分布率有所降低,运转到叶片的比率明显降低。（３）蚕体,蚕砂和蚕茧的镉含量,镉累积量,随着桑叶镉含量的增加而增加。     

Phytogenic compounds from avocado (Persea americana L.) extracts; antioxidant activity,amylase inhibitory activity,therapeutic potential of type 2 diabetes

Diabetes is a worldwide public health disease. Currently, the most effective way to treat diabetes is to mitigate postprandial hyperglycemia by inhibiting carbohydrate hydrolysis enzymes in the digestive system. Plant extracts are rich in bioactive compounds, which can be used in diabetes treatment. This study aims to evaluate the polyphenols content in ethanolic extracts of avocado fruit and leaves (Persea americana Mill.). Additionally, their antioxidant activity using DPPH, while the inhibition ability of α-amylase was examined by reacting different amounts of the extracts with α-amylase compared to acarbose as standard inhibitor. The active compounds were detected in the extracts by LC/MS. The obtained results showed that the leaf extract recorded a significant content of total phenolic compounds compared to the fruit extract (178.95 and 145.7 mg GAE /g dry weight, respectively). The total flavonoid values ??ranged from 32.5 to 70.08 mg QE/g dry weight of fruit and leaves extracts, respectively. Twenty-six phytogenic compounds were detected in leaf and fruit extract by LC/MS. These compounds belong to fatty acids, sterols, triterpenes, phenolic acids, and flavonoids. The antioxidant activity of the extracts is due to the exist of phytogenic compounds, i.e., polyphenols and flavonoids. The antioxidant activity increased in a concentration dependant manner. Avocado fruit extract (1000 µg/mL) scavenged 95% of DPP? while leaf extract rummaged 91.03% of free radicals compared with Vit C and BHT. Additionally, higher α-amylase inhibitory activity was observed in fruit extract than the leaf extract, where the fruit and leaf extract (1000 μg/ml) inhibited the enzyme by 92.13% and 88.95%, respectively. The obtained results showed that the ethanolic extracts of avocado could have a significant impact on human health due to their high content of polyphenols.     

A combined network pharmacology and molecular biology approach to investigate the active ingredients and potential mechanisms of mulberry (Morus alba L.) leaf on obesity

BackgroundAs one of traditional Chinese medicine, mulberry leaf is abundant in diverse active ingredients and widely used for the treatment of metabolic disease and its complications. However, there are a few of reports on its application in the prevention and treatment of obesity. And the molecular mechanism on the anti-obesity of mulberry leaf are unknown till now.PurposeThe present study aimed to evaluate the potential ingredients and targets of mulberry leaf and uncover the anti-obesity mechanisms by using the network pharmacology tactics and verify its effect by biological experiments.Study designActive ingredients and key targets of mulberry leaf, genes related to obesity were screened through public database. Based on the results of network pharmacology, the flavonoids-enriched fraction of mulberry leaf (MLF) was extracted and composition of this fraction was identified. After that, HepG2 cells model of lipid accumulation was established for verifying the effect of MLF and related mechanisms.ResultsA total of 37 active ingredients in mulberry leaf, 192 predicted biological targets and 8813 obesity-related targets were determined, of which 180 overlapping targets might have obvious curative effects on obesity. The networks showed that mulberry leaf might play a role through key targets, such as AKT, MAPK and IL-6, and regulated PI3K-Akt signaling pathway. Based on HPLC-ESI-QQQ-MS analysis, 13 constituents of MLF were identified, including 9 flavonoids. Furthermore, HepG2 cells model of lipid accumulation was established. The results indicated that MLF treatment could down-regulate the secretion of inflammatory cytokines, as well as clearly inhibited lipid droplets formation and alleviated TC, TG, HDL-C and LDL-C levels. Positive effect was observed on hypolipidemic efficacy due to the regulation of PI3K/Akt/Bcl-xl pathway, as indicated by the amelioration of PI3K, Akt and Bcl-xl gene and protein expression.ConclusionThis study firstly systematically disclose the multi-ingredients, multi-targets mechanisms of mulberry leaf on obesity by using network pharmacology approach, and validate in HepG2 cells that the protective effect of MLF against obesity involved both inflammation response and lipid metabolism involving PI3K/Akt/Bcl-xl signaling pathway. It provides indications for further mechanistic research of mulberry leaf and also for the development as a potential candidate for the therapy for obese patients.     

Leaf yield loss assessment due to Macrophomina root rot disease in mulberry gardens of south India

Abstract

Leaf yield loss in mulberry due to Macrophomina root rot disease was assessed in three different states of south India at field level. The highest leaf yield loss recorded was in V-1 variety (34.74%), whereas the lowest leaf yield loss was in K-2 variety (28.54%). However, the leaf yields losses in other varieties viz., MR-2 (32.90%), S-36 (32.06%), RFS-175 (31.75%) and S-13 (29.0%) recorded were medium. The average leaf yield loss was 31.49% due to root rot disease caused by M. phaseolina in mulberry.     

桑叶不同化学成分的降血糖作用

桑叶是一种重要的降血糖药用植物资源。以秋桑叶中提取的六种活性化学成分为材料,研究了六种提取物对四氧嘧啶诱导的糖尿病小鼠的降血糖效果。结果显示,各提取物均有一定的降血糖作用,其中以桑叶总多糖的功效最为显著,桑叶生物碱及桑叶黄酮苷类效果次之。     

Simultaneous determination of the major active components of tea polyphenols in rat plasma by a simple and specific HPLC assay

A simple and specific HPLC assay for simultaneous determination of two major active components (-) epigallocatechin-3-gallate (EGCG), and (-) epicatechin-3-gallate (ECG) of tea polyphenols (TP) in rat plasma was developed and validated. Following addition of resorcinol as internal standard (IS) the analytes were isolated from rat plasma by liquid-liquid extraction with ethyl acetate. The chromatographic separation was achieved on a reversed-phase C18 column using an isocratic mobile phase consisting of 0.1% citric acid+CH(3)CN (86:14, v/v) running at flow rate of 1.5 mL/min. The effluent was monitored at a wavelength of 280 nm. EGCG, ECG and IS were well separated from each other and free from interference from blank plasma and other components in TP as well as metabolites post-dosing. The calibration curve was constructed by plotting peak area ratio of analytes to IS vs. concentration. The method showed good linearity over range of 0.5-300 microg/mL for EGCG and 0.1-60 microg/mL for ECG (r>0.999). The intra- and inter-day precision (R.S.D.) was better than 6 and 12%, respectively. Assay accuracy was better than 94.78% for both compounds. Extraction recovery at QC samples was between 85.73 and 91.93% for EGCG and 79.08 and 86.51% for ECG. The developed method was successfully used to simultaneously measure plasma concentrations of EGCG and ECG after intravenous administration of TP to rats and yielded two typical biexponential decay concentration-time curves.