采用核磁共振/液相色谱-质谱异相关谱方法开展药用植物草棉活性提取物中多组分的同步结构分析研究

基于不完全分离分析策略,建立了NMR/LC-MS异相关谱(NMR/LC-MS HCS)方法对系列混合物样品的LC-MS和NMR谱数据的整合分析,发掘混合物图谱中来自同一组分的LC-MS和NMR谱数据间的内在相关性.以新疆维吾尔民族药用植物草棉(Gossypium herbaceam L.)花瓣的乙醇提取物(AB-8-2...     

瓯柑石油醚提取物的GC-MS分析

为了研究瓯柑的挥发性成分和脂溶性成分, 采用GC-MS分析法进行分离鉴定, 并用面积归一法确定各成分的质量分数. 从瓯柑果肉石油醚提取物中分离鉴定出11种成分, 占总提取物的97.43%, 其主要成分为(Z, Z)-9, 12-十八碳二烯酸(37.46%)、γ-谷甾醇(22.91%);从瓯柑皮石油醚提取物中分离鉴定出10种成分, 占总提取物的99.79%, 其主要成分为柠檬烯(86.39%).     

番石榴叶提取物的鉴定及其抗菌作用

采用纸片扩散法对番石榴叶提取物进行体外抗菌试验研究,以了解其抗菌谱.结果表明番石榴叶提取物中存在有效抗菌成分,其中以乙醇提取物中白念珠菌、甲醇提取物中大肠埃希菌高度敏感.     

溪黄草及其不同提取物的红外光谱研究

采用一维红外光谱、二阶导数谱和二维相关红外光谱对溪黄草原药材及其不同溶剂提取物进行了红外光谱研究。结果表明:溪黄草原药材中含有酯类、芳香类和糖苷类等物质,溪黄草水提物中主要为糖类物质,乙醇提取物中主要为黄酮类物质,石油醚提取物中主要为酯类物质。红外光谱能够快速、简便地提供中药及其提取物中主要化学成分的宏观信息,从而为后续的化学成分分析和中药提取分离方法的改进和优化提供有效参考。     

乌梅二氯甲烷提取物化学成分的GC/MS分析

采用GC-MS联用方法对乌梅二氯甲烷超声提取物和索氏提取物的化学成分分别进行分析。索氏提取物分离了16个峰,经数据库鉴定出13种成分。超声提取物分离了16个峰,经数据库签定出12种成分。     

中药化州橘红乙酸乙酯提取物中微量元素的分布及含量

以乙酸乙酯为溶剂萃取分离了中药化州橘红水煎液的有机成分；测定了提取物中元素的分布及含量，分析了不同微量元素在提取物中富集程度的差别。并由此说明了中药微量元素与中药所含的有机成分是有密切关系的。     

大豆中Glycinol的分离与鉴定

 Glycinol是大豆中主要的植物抗毒素和植物雌激素glyceollins的直接前体,是研究大豆中异戊烯转移酶必需的重要底物。该研究以新鲜大豆为原料，采用硝酸银诱导大豆子叶使glycinol 在其中累积,再用甲醇提取，经制备高效液相色谱分离获得单一成分;经液相色谱/四极杆-飞行时间串联质谱及紫外光谱鉴定,并与标准品对照,确定所获得的单一成分为glycinol。采用该分离方法可从390 g鲜重的大豆中分离得到100 mg高纯度的glycinol。该研究提供了一种分离纯化和较大量制备glycinol 的方法。     

超高效液相色谱法测定大黄与铜离子络合能力

中药肝豆汤治疗铜代谢障碍疾病具有疗效显著、副作用小的优势,但由于缺乏评价铜离子(Cu²⁺)络合能力的技术手段,限制了肝豆汤配位活性成分的筛查和发现。为评价复方肝豆汤中主药大黄对铜离子的络合能力,该文通过优化大黄活性成分与铜离子配位反应条件,建立了超高效液相色谱法(UHPLC)测定中药大黄与铜离子络合能力的方法。样品经Agilent Eclipse Plus C18色谱柱(50 mm×2.1 mm, 1.8μm)分离,以甲醇-0.1%(v/v)磷酸水溶液为流动相进行梯度洗脱,柱温30℃,流速为0.3 mL/min,检测波长为254 nm,进样体积为5μL。研究通过UHPLC分析比较大黄与铜离子配位反应前后色谱峰变化,超高效液相色谱四极杆-飞行时间质谱(UPLC-Q-TOF-MS)鉴定大黄提取物中配位活性成分,计算大黄提取物与铜离子配位反应前后其色谱峰面积变化率来评价大黄中活性成分与Cu²⁺的络合能力。结果显示,大黄提取物与Cu²⁺在pH为9的体系中配位反应12 h达到平衡;鉴定出大黄提取物中20种主要成分,根据Cu     

红芪药材防治肝纤维化的谱效关系

研究了红芪药材的高效液相色谱(HPLC)指纹图谱与抗肝纤维化作用之间的谱效关系。采用四氯化碳(CCl4)-花生油(体积比为4∶6)皮下注射(0.1 mL/10 g,1次/5 d)致小鼠肝纤维化,同时灌胃给予红芪各不同溶剂提取物(10 g原药材/kg,1次/d),5周后测定与肝纤维化有关的各血清指标及肝重指数,结果乙醇提取物药效最佳。运用灰色关联度和偏最小二乘法研究了不同产地红芪药材乙醇提取部位的HPLC指纹图谱与抗肝纤维化药效之间的谱效相关性。结果红芪乙醇提取物中多数成分与抗肝纤维化药效均有高度关联性(0.8),表明该药效是红芪中所有成分共同作用的结果;同时鉴定出腺苷、毛蕊异黄酮及芒柄花苷3种成分,其中腺苷及毛蕊异黄酮对药效有较大贡献。     

金银花的微乳液薄层色谱分离方法研究

以CTAB/正丁醇/正庚烷/水微乳液作为流动相,在聚酰胺薄层上用薄层色谱法满意地分离了金银花提取物中的各类成分,并用薄层色谱扫描仪进行荧光扫描,建立了金银花的微乳液薄层色谱扫描图谱,为金银花指纹图谱的研究奠定了基础.     

Nuclear magnetic resonance and liquid chromatography-mass spectrometry combined with an incompleted separation strategy for identifying the natural products in crude extract

NMR and LC-MS combined with an incompleted separation strategy were proposed to the simultaneous structure identification of natural products in crude extracts, and a novel method termed as NMR/LC-MS parallel dynamic spectroscopy (NMR/LC-MS PDS) was developed to discover the intrinsic correlation between retention time (Rt), mass/charge (m/z) and chemical shift (δ) data of the same constituent from mixture spectra by the co-analysis of parallelly visualized multispectroscopic datasets from LC-MS and ¹H NMR. The extracted ion chromatogram (XIC) and ¹H NMR signals deriving from the same individual constituent were correlated through fraction ranges and intensity changing profiles in NMR/LC-MS PDS spectrum due to the signal amplitude co-variation resulted from the concentration variation of constituents in a series of incompletely separated fractions. NMR/LC-MS PDS was applied to identify 12 constituents in an active herbal extract including flavonol glycosides, which was separated into a series of fractions by flash column chromatography. The complementary spectral information of the same individual constituent in the crude extract was discovered simultaneously from mixture spectra. Especially, two groups of co-eluted isomers were identified successfully. The results demonstrated that NMR/LC-MS PDS combined with the incompleted separation strategy achieved the similar function of on-line LC-NMR-MS analysis in off-line mode and had the potential for simplifying and accelerating the analytical routes for structure identification of constituents in herbs or their active extracts.     

In vitro toxicity, antiplatelet and acetylcholinesterase inhibition of Buddleja thyrsoides Lam. leaves

Alzheimer's disease (AD) is a neurodegenerative disorder resulting in impaired memory and behaviour of remarkable socio-economic impact. A decrease in cholinergic activity is a key event in the biochemical of AD. Buddleja thyrsoides is a plant widely distributed in Southern parts of South America. In Brazilian traditional medicine, the infusion of its leaves and flowers is used for the treatment of bronchitis and cough. Crude ethanolic (70%) extract and fractions (dichloromethane, ethyl acetate and n-butanolic) were investigated regarding their toxicities in?vitro and antiplatelet action. The enzyme acetylcholinesterase inhibition was evaluated to study the crude extract. The crude extract and fractions were evaluated by means of Brine Shrimp Lethality test and they showed low activities with LC(50) values 1698, 2818, 2187 and 3672?μg?mL(-1) for dichloromethane, ethyl acetate, n-butanolic fractions and crude extract, respectively. Buddleja thyrsoides presented great antiplatelet action. The IC(50) values obtained for crude extract and dichloromethane, ethyl acetate and n-butanolic fractions were 361.29, 354.23, 368.75 and 344.30, respectively, while the IC(50) for the standard AAS was 257.01?μg?mL(-1). The crude extract showed an inhibition of 22.8% of the acetylcholinesterase enzyme in 24?h.     

Antibacterial, antifungal, antispasmodic and Ca++ antagonist effects of Caesalpinia bonducella

Caesalpinia bonducella F. (Leguminosae) has been used as a folk medicine for a variety of ailments. The crude extract of C. bonducella and its fractions were studied for antibacterial, antifungal, antispasmodic and Ca++ antagonistic properties. The strongest antibacterial effect was displayed by the n-butanol (72%) and ethyl acetate (80%) fractions, followed by the crude extract (46% and 42%), against Escherichia coli and Bacillus subtilis, respectively. The plant extract and its fractions showed mild to excellent activity in antifungal bioassays, with maximum antifungal activity against Candida glaberata (80%) and Aspergillus flavus (70%) by the n-butanol and chloroform fractions, followed by the crude extract (70% and 65%). Caesalpinia bonducella extract caused concentration-dependent inhibition of spontaneous and high K+ (80 mM)-induced contractions of isolated rabbit jejunum preparations, similar to that caused by Verapamil. These results indicate that C. bonducella exhibits antibacterial, antifungal, spasmolytic and Ca++ channel blocking actions.     

Antioxidative properties of defatted dabai pulp and peel prepared by solid phase extraction

Solid phase extraction (SPE) using Sep-Pak^? cartridges is one of the techniques used for fractionation of antioxidant compounds in waste of dabai oil extraction (defatted dabai parts). The aim of this study was to determine the phenolic compounds and antioxidant capacity in crude extracts and several SPE fractions from methanolic extract of defatted dabai pulp and peel. Based on SPE, Sep-Pak^? cyanopropyl and C₁₈ cartridges were used to fractionate the antioxidant-rich crude extracts into water and methanolic fractions. Analyzed using LC-MS, flavonoids, anthocyanins, saponin derivatives and other unknown antioxidative compounds were detected in the defatted dabai crude extracts and their SPE fractions. Anthocyanins were the major phenolic compounds identified in the defatted dabai peel and detected in most of the SPE fractions. Methanolic fractions of defatted dabai parts embraced higher total phenolics and antioxidant capacity than water fractions. This finding also revealed the crude extracts of defatted dabai peel have the most significant antioxidant properties compared to the methanolic and water fractions studied. The crude extract of defatted dabai parts remain as the most potent antioxidant as it contains mixture of flavonoids, anthocyanins and other potential antioxidants.     

Antifertility activity of Piper longum Linn. in female rats

The crude extract, its different fractions and the major pure compound from the active fraction of the powdered fruits of Piper longum were studied for the antifertility effect in female rats. The crude extract and its hexane fraction exhibited 100 and 86% efficacy respectively (days 1-7 post-coitum (p.c.) schedule). On the other hand, 1-butanol soluble, 1-butanol insoluble and chloroform fractions were inactive.     

Acute toxicity and hepatotoxicokinetic studies of Tamarindus indica extract

Tamarindus indica is widely used as a food and beverage and in traditional medicine. The apparent lack of dose standardization in herbal medicine necessitates the evaluation of the lethality T. indica on Artemia salina nauplii and chicken embryos via in vitro and in vivo techniques. Furthermore, hepatotoxicokinetics of the crude extract and fractions on Wister rats was also assessed. At concentrations of 200, 20 and 2 μg/mL, crude extract and fractions showed brine shrimp death percentages ranging from 86.70% to 3.30% and the sub-fractions showed death percentage ranges of 46.70% to 3.30%. Calculated LD?? values ranged from 832 μg/mL to 5,019 μg/mL. Dosing Wister rats with 25% and 50% concentration of LD?? determined for crude extract and fractions on chicken embryos showed an elevation in the ALT and AST levels in the serum. Brine shrimps and chicken embryos showed a positive correlation, with R2 values of 0.541 and 0.588 (P ≤ 0.05) for fractions and subfractions, respectively, as media for the lethality assay. Dose standardization in folk herbal medicine is imperative as T. indica used as food and medicine has been shown to be toxic at high doses. Brine shrimp and chicken embryos may be comparably used as medium for toxicity assay.     

Anti-inflammatory and antiproliferative activities of Ixora brevifolia Benth. (Rubiaceae)

The crude extract and fractions from the branches of Ixora brevifolia, a tree found in the Brazilian Cerrado, were tested for anti-inflammatory and in vitro antiproliferative effects. The crude extract and n-hexane fraction exhibited significant inhibition of ear oedema in mice, while n-hexane-precipitated and chloroform fractions strongly inhibited the myeloperoxidase activity in ear tissue. The n-hexane and n-hexane-precipitated fractions showed strong growth inhibition for glioma cell line and the hydromethanolic fraction inhibited the growth of leukaemia cell line.     

Isolation of quercetin and mandelic acid from <Emphasis Type="Italic">Aesculus indica</Emphasis> fruit and their biological activities

Background

In this study Aesculus indica fruit was subjected to isolation of phytochemicals. Two antioxidants quercetin and Mandelic acid were isolated in pure state. The free radical scavenging and acetyl choline esterase inhibitory potential of the crude extract and sub fractions were also determined.

Results

The antioxidant capacity of crude extract, fractions and isolated compounds were determined by DPPH and ABTS methods. Folin-Ciocalteu reagent method was used to estimate the total phenolic contents and were found to be 78.34?±?0.96, 44.16?±?1.05, 65.45?±?1.29, 37.85?±?1.44 and 50.23?±?2.431 (mg/g of gallic acid) in crude extract, ethyl acetate, chloroform, n-hexane and aqueous fractions respectively. The flavonoid concentration in crude extract, ethyl acetate, chloroform, n-hexane and aqueous fraction were; 85.30?±?1.20, 53.80?±?1.07, 77.50?±?1.12, 26.30?±?1.35 and 37.78?±?1.25 (mg/g of quercetin) respectively. The chloroform fraction was more potent against enzymes, acetyl choline esterase and butyryl choline esterase (IC₅₀?=?85 and 160 μg/ml respectively). The phenolic compounds in the crude extract and fractions were determined using HPLC standard method. Chlorogenic acid, quercetin, phloroglucinol, rutin, mandelic acid and hydroxy benzoic acid were detected at retention times 6.005, 10.062, 22.623, 30.597, 35.490 and 36.211 in crude extract and different fractions. The ethyl acetate fraction was rich in the targeted compounds and was therefore subjected to column isolation. The HPLC chromatogram of isolated compounds showed single peak at specified retention times which confirms their isolation in pure state. The isolated compounds were then characterized by FTIR and NMR spectrophotometric techniques.

Conclusion

The Aesculus indica fruit extracts showed antioxidant and anticholine esterase inhibitory potentials. Two bioactive compounds were isolated in the pure form ethyl acetate fraction. From results it was concluded that the fruit of this plant could be used to minimize oxidative stress caused by reactive oxygen species.

     

Short and long-term effects of Baccharis articulata on glucose homeostasis

In this study, the in vivo effect of the crude extract and n-butanol and aqueous residual fractions of Baccharis articulata (Lam.) Pers. on serum glucose levels, insulin secretion and liver and muscle glycogen content, as well as in vitro action on serum intestinal disaccharidase activity and albumin glycation were investigated. Oral administration of the extract and fractions reduced glycemia in hyperglycemic rats. Additionally, the n-butanol fraction, which has high flavonoids content, stimulated insulin secretion, exhibiting an insulinogenic index similar to that of glipizide. Also, the n-butanol fraction treatment significantly increased glycogen content in both liver and muscle tissue. In vitro incubation with the crude extract and n-butanol and aqueous residual fractions inhibited maltase activity and the formation of advanced glycation end-products (AGEs). Thus, the results demonstrated that B. articulata exhibits a significant antihyperglycemic and insulin-secretagogue role. These effects on the regulation of glucose homeostasis observed for B. articulata indicate potential anti-diabetic properties.