云南丹参资源植物的成分测定与质量评价

丹参为唇形科鼠尾草属（Salvia L．）植物丹参（Salvia mihiorrhiza Bge．）的干燥根及根茎,具有祛瘀止痛、活血通经、清心除烦之功效。丹参的有效成分为脂溶性的二萜醌类和水溶性的酚酸类成分,均具有心血管方面的药理作用。其有效成分含量的测定主要有HPLC法、紫外分光光度法等。云南的鼠尾草属植物（丹参）资源较为丰富,作者采用高效液相色谱法和紫外分光光度法测定了采自云南的鼠尾草属7种1变种的丹参酮类成分和丹酚酸类成分,以期为云南产丹参品质评价及资源利用提供更多的科学依据。     

经典名方温胆汤物质基准总黄酮、柚皮苷和新橙皮苷含量测定

目的：研究并建立紫外分光光度法（UV）和高效液相色谱法（HPLC）分别测定温胆汤物质基准中总黄酮、柚皮苷和新橙皮苷含量。方法：以柚皮苷为对照品，采用紫外-可见分光光度法测定总黄酮的含量并采用HPLC法测定柚皮苷和新橙皮苷的含量。结果：采用直接测定法测定总黄酮类成分在284 nm处有较强吸收，柚皮苷在20.22～80.88μg/mL范围内与吸光度呈良好的线性关系，平均加样回收率为98.33%。柚皮苷和新橙皮苷分别在0.01～0.51 mg/mL和0.01～0.50 mg/mL范围内有良好的线性关系，精密度、稳定性、重复性的RSD均小于5.0%，加样回收率分别为97.11%和102.72%；温胆汤物质基准中总黄酮的平均含量为15.03%，柚皮苷、新橙皮苷的平均含量分别为22.78 mg/g和9.75 mg/g。结论：建立的温胆汤物质基准总黄酮、柚皮苷和新橙皮苷含量测定方法简单、稳定、重复性好，为温胆汤及其相关制剂进一步研究和开发提供实验基础。     

拳卷地钱中黄酮类化合物的含量测定

本文建立了拳卷地钱中黄酮类化合物的含量测定方法。采用改进的铝盐显色分光光度法测定拳卷地钱总黄酮含量;采用反相高效液相色谱法测定拳卷地钱黄酮类化合物中芹菜素、槲皮素和木犀草素的含量。分光光度法表明：总黄酮含量在1．6～16μg/mL范围内,线性关系良好,加标回收率在97．61～101．59％,相对标准偏差0．16％;反相高效液相色谱法表明平均加标回收率96．79～101．13％,相对标准偏差0．66～1．52％。本方法用于拳卷地钱中黄酮类化合物含量的测定准确可靠,结果令人满意。     

UPLC-MS/MS法同时测定毕氏海蓬子中13个功能成分

利用超高效液相色谱-串联质谱（UPLC-MS/MS）法同时测定毕氏海蓬子（Salicornia bigelovii,SB）中没食子儿茶素、表没食子儿茶素、隐绿原酸、绿原酸、新绿原酸、儿茶素、表儿茶素、异绿原酸-B、芦丁、异槲皮苷、褪黑素、山柰酚-3-O-芸香糖苷、槲皮苷13个功能成分的含量。样品利用甲醇提取,过0.22 μm滤膜后上机检测。采用C18 色谱柱 （100 mm × 2.1 mm,1.7 μm）进行分离,以0.1%甲酸水溶液-甲醇为流动相,流速0.3 mL/min,柱温（35±5）℃,采用电喷雾离子源（ESI）、多反应监测（MRM）进行检测。结果表明：13个功能成分在各自的线性范围内线性关系良好,相关系数R2均大于0.99,该方法的检测限（LOD）和定量限（LOQ）分别为1.0~20.0 μg/kg、2.0~50.0 μg/kg,平均加标回收率在89.10%~109.51%范围内。LC-MS/MS法测定毕氏海蓬子功能成分快速灵敏,准确可靠,可用于毕氏海蓬子中功能成分的检测。     

拳参中没食子酸和总鞣质的含量测定

该研究采用高效液相色谱法和紫外分光光度法分别对拳参中的主要成份———没食子酸以及总鞣质进行含量测定。其中,用高效液相色谱法测定没食子酸的含量,HPLC 条件为 Hypersil GOLD Phenyl 柱(250 mm ×4.6 mm,5μm),甲醇－0.1％磷酸梯度洗脱,流速为1.0 mL?min-1,检测波长为276 nm。用紫外分光光度法测定总鞣质的含量,采用2010版《中国药典》一部附录 XB 中鞣质含量测定方法,用干酪素沉淀鞣质,通过干酪素沉淀前后鞣质的变化来确定样品中总鞣质的含量。结果表明：没食子酸在0.051~1.02μg 范围内呈现良好的线性关系,平均加样回收率分别为99.1％、100.3％、101.9％,RSD 分别为2.1％、0.8％、2.0％;总鞣质在2.09~10.48μg 范围内呈现良好线性关系,平均加样回收率分别为102.2％、100.2％、102.1％,RSD 分别为1.3％、1.4％、1.0％。利用上述方法,还测定了贵州贵阳和四川成都各3批样品中没食子酸和总鞣质的含量差异,发现成都样品中的没食子酸与总鞣质均大于贵阳拳参中的含量,初步分析导致这一结果的原因可能与拳参的生长海拔有关,而成都的海拔范围更适合拳参的生长。上述两种方法快速、简单、重现性好,可以作为拳参中没食子酸和总鞣质的含量测定方法,而该方法也是首次被用来同时测定拳参中没食子酸和总鞣质的含量,为今后的地方药典标准甚至《中国药典》标准的补充和完善提供了重要依据。     

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.     

紫外可见分光光度法测定杜仲绿原酸含量的方法研究

目前,测定绿原酸含量的方法很多,常采用毛细管电泳法、ＨＰＬＣ法、可见分光光度法和纸层析-紫外可见分光光度法等[1,2,3,4]。但样品处理均采用索氏回流提取,后经层析分离洗脱等步骤,时间长、效率低。本文采用紫外可见分光光度法测定绿原酸含量,只需超声波提取一小时,不经...     

黄山贡菊花、叶中总黄酮及绿原酸动态积累研究

本文分别采用紫外分光光度法和HPLC法对贡菊花及叶中总黄酮及绿原酸的含量进行分析,研究不同生长时期黄山贡菊花和叶中总黄酮与绿原酸动态积累规律。结果表明:黄山贡菊花中总黄酮和绿原酸含量在中花期均达到最高值,而在采摘期内变化较大;黄山贡菊叶中总黄酮和绿原酸含量随着生长期的延长呈现逐渐上升趋势,并在生长后期总黄酮和绿原酸含量分别出现109.30和7.87 mg/g的峰值。     

四种山蓝加工方法的比较

用高效液相色谱法和分光光度法测定了四种加工方法所得山蓝加工提取物中有效成分含量 ,结果表明 ,经阴干 ,晒干 ,乙醇浸泡几种传统加工方法加工的山蓝中紫蓝素含量分别为 31 1、4 5 6、4 72 mg/1 0 0 g(干重 ) ,而采用避氧热处理加工的山蓝中 ,紫蓝素含量达 1 .73g/1 0 0 g。用传统加工方法对山蓝进行加工 ,有效成分含量低 ,制约了山蓝的利用价值 ,而避氧热处理能大大提高有效成分含量 ,提升山蓝的品质。研究结果对山蓝的加工、炮制和开发利用有重大意义。     

Antioxidant Sources from Leaves of Russian Dandelion

Taraxacum kok‐saghyz (TKS) is a dandelion species native to Kazakhstan, Uzbekistan and north‐west China, considered as a promising alternative source of natural rubber from its roots. The aim of this study was to investigate the possible exploitation of TKS leaves, a rubber byproduct, as a source of phenolic compounds with antioxidant properties for potential applications in forage, nutraceutical and pharmacological fields. Two accessions (TKS016, TKS018) grown under Mediterranean conditions of Sardinia were evaluated at vegetative and flowering stages. The leaves of TKS018 had the highest antioxidant capacity (19.6 mmol trolox equivalent antioxidant capacity 100 g^?1), total phenolic (106.4 g gallic acid equivalent kg^?1), tannic phenolics (58.5 g gallic acid equivalent kg^?1) and total flavonoid contents (22.9 g catechin equivalent kg^?1). At both phenological stages, TKS016 showed significantly lower values than TKS018 in 1,1‐diphenyl‐2‐picrylhydrazyl (DPPH), total phenolic and tannic phenolics. Six individual molecules were identified, namely chlorogenic, cryptochlorogenic, caffeic, sinapic, chicoric and 3,4‐dimethoxycinnamic acids. Chicoric (8.53–10.68 g kg^?1 DW) and chlorogenic acids (4.18–7.04 g kg^?1 DW) were the most abundant. TKS leaves represent a valuable source of chicoric acid with potential application as antioxidant to be used as herbal medicine and nutrition for production of healthy food/feed.     

蔓三七叶中分离绿原酸和异绿原酸及其抗氧化活性研究

以蔓三七叶为原料,制备其中的绿原酸和异绿原酸A、B和C,并对它们进行结构鉴定;同时,采用DPPH法、水杨酸比色法、邻苯三酚自氧化法来对其进行体外抗氧化活性研究。结果表明,分离制备的绿原酸和异绿原酸A、B和C质量分数分别为96. 8%、98. 2%、97. 6%和98. 7%。在0. 5～10μg/mL范围内,绿原酸和异绿原酸A、B和C对DPPH和羟基自由基表现出了较好的清除作用,随浓度升高而逐渐增大;但它们对超氧自由基的清除作用时,浓度从5μg/mL升高到100μg/mL时,清除作用都未出现明显的剂量依赖性,各样品与阳性对照组(VC)相比较而言,其清除超氧自由基的作用明显较弱。本文揭示了蔓三七叶中的绿原酸和异绿原酸A、B和C具有不同的抗氧化效果。     

不同甜叶菊品种叶中绿原酸类成分的比较研究

该文以14个扦插培育的甜叶菊品种叶为材料,从8种不同型号的树脂中筛选出一种合适的大孔吸附树脂对甜叶菊叶中绿原酸类成分进行纯化前处理,采用HPLC法对不同甜叶菊品种叶中所含绿原酸类成分进行比较分析。结果表明:(1)在8种不同型号的树脂中,XAD~(-1)6对甜叶菊叶中绿原酸类成分吸附-解析性能最佳。(2)经优化,上样液浓度1.20 mg·mL~(-1)、样品溶液pH 3、解析液乙醇体积分数70%时XAD~(-1)6树脂对甜叶菊叶中绿原酸类成分具有较好的纯化效果。(3) HPLC检测分析表明,在14个品种中共检测出新绿原酸、绿原酸、隐绿原酸、异绿原酸B、异绿原酸A、异绿原酸C六种绿原酸类成分,其中主要成分均为异绿原酸A、绿原酸、异绿原酸C,而在品种3、5、13、14中没有检测出异绿原酸B。(4) 14个品种中6个绿原酸类成分的含量分别为异绿原酸A 20.55~54.3 mg·g~(-1)、绿原酸17.96~32.93 mg·g~(-1)、异绿原酸C 4.15~19.49 mg·g~(-1)、新原酸0.61~4.61 mg·g~(-1)、隐绿原酸0.52~3.11 mg·g~(-1)、异绿原酸B 0.0~3.17 mg·g~(-1),6种绿原酸类成分总量为43.9~97.8 mg·g~(-1)。可见,不同品种甜叶菊叶中绿原酸类成分含量有明显差异,富含绿原酸类成分的甜叶菊品种可用于开发获取绿原酸类物质。     

Chemical Characterization and Multidirectional Biological Effects of Different Solvent Extracts of Arum elongatum: in Vitro and in Silico Approaches

Arum elongatum (Araceae) is widely used traditionally for the treatment of abdominal pain, arterial hypertension, diabetes mellitus, rheumatism and hemorrhoids. This study investigated the antioxidant properties, individual phenolic compounds, total phenolic and total flavonoid contents (HPLC/MS analysis), reducing power and metal chelating effects of four extracts obtained from A. elongatum (ethyl acetate (EA), methanol (MeOH), methanol/water (MeOH/water) and infusion). The inhibitory activity of the extracts were also determined against acetylcholinesterase, butyrylcholinesterase, tyrosinase, amylase and glucosidase enzymes. The MeOH/water extracts contained the highest amount of phenolic contents (28.85 mg GAE/g) while the highest total flavonoid content was obtained with MeOH extract (36.77 mg RE/g). MeOH/water demonstrated highest antioxidant activity against DPPH⋅ radical at 38.90 mg Trolox equivalent per gram. The infusion extract was the most active against ABTS⁺⋅ (133.08 mg TE/g). MeOH/water extract showed the highest reducing abilities with the CUPRAC value of 102.22 mg TE/g and the FRAP value of 68.50 mg TE/g. A strong metal chelating effect was observed with MeOH/water extract (35.72 mg EDTAE/g). The PBD values of the extracts ranged from 1.01 to 2.17 mmol TE/g. EA extract displayed the highest inhibitory activity against AChE (2.32 mg GALAE/g), BChE (3.80 mg GALAE/g), α-amylase (0.56 mmol ACAE/g) and α-glucosidase (9.16 mmol ACAE/g) enzymes. Infusion extract was the most active against tyrosinase enzyme with a value of 83.33 mg KAE/g. A total of 28 compounds were identified from the different extracts. The compounds present in the highest concentration were chlorogenic acids, 4-hydroxybenzoic acid, caffeic acid, p-coumaric acid, ferulic acid, isoquercitrin, delphindin 3,5-diglucoside, kaempferol-3-glucoside and hyperoside. The biological activities of A. elongatum extracts could be due to the presence of compounds such as gallic acid, chlorogenic acids, ellagic acid, epicatechin, catechin, kaempferol, 4-hydroxybenzoic acid, caffeic acid, p-coumaric acid, ferulic acid, quercetin, isoquercitrin, and hyperoside. Extracts of A. elongatum showed promising biological activities which warrants further investigations in an endeavor to develop biopharmaceuticals.     

环渤海湾地区连作苹果园土壤中酚酸类物质变化

分析了山东昌邑、栖霞、蓬莱,辽宁大连、抚宁、绥远,河北昌黎、青县等地苹果园连作土壤中酚酸物质的组成和含量,结果表明:连作障碍发生的苹果园土壤中酚酸类物质的组成和含量在不同地区、不同土层厚度间存在显著差异。苹果园土壤中酚酸类物质的含量从春季到秋季随时间的延长逐渐减少,但过程缓慢。且非连作园土壤中酚酸类物质含量显著少于连作土。连作土壤中酚酸物质的组成和含量在不同季节间差异显著,尤其春季土壤中酚酸物质的种类与夏季和秋季显著不同。这可能是因为植物在不同季节分泌的酚酸类物质种类和含量有差别.不同土层中酚酸物质的分布因季节不同而有显著差异。夏季土壤中酚酸物质主要分布在浅层土壤中,而秋季则主要分布在深层土壤中。这可能是由于浇水等果园管理措施和自然降水对土壤中的酚酸物质产生的淋溶作用,使得大量的酚酸物质向深层土壤运动,最终造成了秋季果园连作土中随土层加深而酚酸物质含量增加的现象。不同地区苹果园连作土中,对羟基苯甲酸、(+)-儿茶素、咖啡酸、阿魏酸含量与非连作土无显著差异,而焦性没食子酸、绿原酸和根皮苷显著高于非连作土。焦性没食子酸、绿原酸和根皮苷可能是引起山东、辽宁、河北地区苹果园连作障碍的关键酚酸物质。     

Extraction of Functional Compounds from Tarragon (Artemisia dracunculus L.) by Deep Eutectic Solvents at Different Properties

In this study, it was aimed to examine the capacity of deep eutectic solvents (DESs) with different contents to extract bioactive compounds from tarragon (Artemisia dracunculus L.) plant. For this reason, the total phenolic-flavonoid content, total proanthocyanidin content and antioxidant/antimicrobial activities of the prepared DES extracts were investigated, as well as the individual phenolic and individual amino acid profiles. According to the results, DES10 had the highest efficiency in terms of its capacity to extract individual phenolics (approximately 59 mg/100 g) and individual amino acids (approximately 2500 mg/kg), and also gave a higher yield compared to ethanol (approximately 44 mg/100 g for individual phenolics and about 19810 mg/kg for individual amino acids) and methanol (approximately 58 mg/100 g for individual phenolics and approximately 21430 mg/kg for individual amino acids). However, the total phenolic content, total flavonoid content and antioxidant activity values of DES extracts were determined between 59.09–77.50 mg GAE/100 g, 28.68–45.55 mg GAE/100 g and 42.96–146.86 mg TE/100 g, respectively. Therefore, it can be recommended to use these green solvents, which are known as environmentally friendly, as an alternative to organic solvents in the process of preparing extracts of this important medicinal plant in different areas.     

菹草提取物总黄酮、总酚酸和抗氧化活性含量研究

研究菹草提取物的有效成分,测定其总黄酮含量及总酚酸含量,并确定它的乙酸乙酯和石油醚部位的抗氧化活性成分。采用紫外可见分光光度法,以芦丁和没食子酸作为控制材料,测定菹草的石油醚部位及乙酸乙酯部位中总黄酮和总酚酸含量;采用清除DPPH自由基能力测定法、测定总还原能力铁氰化钾法和水杨酸捕捉羟基自由基法来对菹草各组分提取物的抗氧化能力进行研究。菹草中有一定量的黄酮类化合物和酚酸类化合物存在,不同溶剂提取的部位所含有的总黄酮和总酚酸含量是有差异的,石油醚部位的总黄酮含量要高于乙酸乙酯部位的总黄酮含量,并且总酚酸的含量亦是如此;石油醚部位和乙酸乙酯部位的提取物具有清除DPPH、羟基自由基和还原Fe^3+的能力,各部位清除DPPH、羟基自由基的能力和还原Fe^3+的能力随着样品浓度的增大而增大,且乙酸乙酯部位的测定结果均低于石油醚部位。     

Antistaphylococcal and biofilm inhibitory activities of gallic,caffeic, and chlorogenic acids

Staphylococcus aureus is a Gram-positive pathogen which is able to form biofilms, exhibiting a more pronounced resistance to antibiotics and disinfectants. The?hurdles posed in eradicating biofilms?have driven the search for new compounds able to fight these structures. Phenolic compounds constitute one of the most numerous and ubiquitous group of plant secondary metabolites with many biological activities. The aim of the present work was to study the potential antimicrobial and antibiofilm properties of gallic, caffeic, and chlorogenic acids against S. aureus as well to elucidate its mechanism of action. It was concluded that the phenolic acids studied in this work?have antistaphylococcal properties. For instance, gallic acid is able to influence the adhesion properties of S. aureus. The phenolic acids tested were also able to inhibit the production of α-hemolysin by this microorganism, with the exception of chlorogenic acid. Regarding its mechanism of action, caffeic acid interferes with the stability of the cell membrane and with the metabolic activity of the cells of S. aureus.     

Overexpression of a chrysanthemum transcription factor gene <Emphasis Type="Italic">DgNAC1</Emphasis> improves drought tolerance in chrysanthemum

Phenolic acids and tanshinones are two groups of pharmaceutical components present in Salvia miltiorrhiza Bunge. Methyl jasmonate (MeJA) has been reported to influence the accumulation of both phenolic acids and tanshinones in S. miltiorrhiza hairy roots. However, there is currently a lack of information regarding the comparison of how these two groups of bioactive compounds in S. miltiorrhiza respond to MeJA under the same conditions. In the present study, the effect of 100 µM MeJA on the biosynthesis of phenolic acids and tanshinones in S. miltiorrhiza hairy roots was investigated. The results showed that MeJA dramatically induced the accumulation of five different phenolic acids, especially rosmarinic acid and salvianolic acid B, which reached their highest contents at day 3 (20.3 mg/g DW, 1.5-fold of control) and day 6 (47.49 mg/g DW, 2.5-fold of control), respectively. The total production of phenolic acids was induced by as much as 3.3-fold of the control (day 9 after treatment), reaching 357.5 mg/L at day 6. However, tanshinone I was almost unaffected by MeJA treatment, and the accumulation of tanshinone IIA was inhibited. Furthermore, cryptotanshinone and dihydrotanshinone I were moderately induced by MeJA. The gene expression results indicated that MeJA probably induced the whole pathways, especially the tyrosine-derived pathway and the methylerythritol phosphate pathway, and finally resulted in the increased production of these metabolites. This study will help us to further understand how the different biosynthetic mechanisms of phenolic acids and tanshinones respond to MeJA and provide a reference for the future selection of elicitors for application to improving the production of targeted compounds.     

Phenolic compounds of Sorghum vulgare in response to Sclerotium rolfsii infection

Abstract

Identification of individual phenolic acids of Sorghum vulgare Pers. cv. M.P. after interaction with Sclerotium rolfsii Sacc. using high performance liquid chromatograph (HPLC) showed the presence of phenolics namely tannic, gallic, ferulic, chlorogenic and cinnamic acids in varying amounts. After 72 h inoculation with S. rolfsii, a maximum amount of ferulic acid (166.6 µg g^?1 fresh wt) was present in the collar of inoculated plants, followed by leaves and roots and its level decreased gradually with time. Similarly, the presence of chlorogenic acid was traced after 48 h, while that of cinnamic acid was traced after 72 h of inoculation. Reddish-brown pigmentation at the collar region of inoculated plants was also observed along with the high content of tannic acid. Among other phenolics, the presence of gallic acid was recorded consistently and maximum accumulation (139.3 µg g^?1 fresh wt) was noticed at the zone of interaction (collar region) after 72 h of inoculation. In contrast, maximum lignin deposition was observed at collar region after 96 h of inoculation. Induction of phenolic acids in S. vulgare along with the lignin deposition and red pigmentation at collar region is considered a key biomarker in the non-host-pathogen interaction in the S. valgare–S. rolfsii pathosystem.