越鞠丸石油醚提取物的气相色谱-质谱分析

采用气相色谱-质谱联用技术对中药复方越鞠丸石油醚提取物中的成分进行鉴定,共鉴定出60种成分,并采用峰面积归一化法确定了各成分的相对含量。     

Current analytical methods to study plant water extracts: the example of two mushrooms species, Inonotus hispidus and Sparassis crispa

The analytical study of two hot-water extracts from the mushrooms Inonotus hispidus (Bull.) P. Karst and Sparassis crispa Wulf.:Fr was performed by NMR, HPLC-PAD-MS and GC-MS. The simultaneous use of different analytical techniques highlighted the diverse classes of natural products contained in these extracts. This study describes an attempt to adapt a useful phytochemical method to the direct investigation of plant water extracts, which represent the typical traditional manner for the administration of natural remedies. The heritage concerning plant processing procedures, known as traditional pharmaceutical knowledge, could play an important role in future research on medicinal species. This kind of study could be used as an update for current and future perspectives in this research field.     

无叶假木贼和盐爪爪提取物的抗菌活性

无叶假木贼和盐爪爪地上部分乙醇提取物、不同溶剂萃取部分对供试病原细菌和真菌均表现出较好的抑制作用,其乙酸乙酯萃取部分和正丁醇萃取部分的抗菌活性明显强于石油醚萃取部分和水部分。结果表明,无叶假木贼中抗菌活性成分主要为极性中等的化合物,且很可能是具弱碱性、易与酸成盐的生物碱类。盐爪爪中抗菌活性成分同样为极性中等的化合物。     

Screening of plant extracts for antimicrobial activity against bacteria and yeasts with dermatological relevance

There is cumulative resistance against antibiotics of many bacteria. Therefore, the development of new antiseptics and antimicrobial agents for the treatment of skin infections is of increasing interest. We have screened six plant extracts and isolated compounds for antimicrobial effects on bacteria and yeasts with dermatological relevance. The following plant extracts have been tested: Gentiana lutea, Harpagophytum procumbens, Boswellia serrata (dry extracts), Usnea barbata, Rosmarinus officinalis and Salvia officinalis (supercritical carbon dioxide [CO2] extracts). Additionally, the following characteristic plant substances were tested: usnic acid, carnosol, carnosic acid, ursolic acid, oleanolic acid, harpagoside, boswellic acid and gentiopicroside. The extracts and compounds were tested against 29 aerobic and anaerobic bacteria and yeasts in the agar dilution test. U. barbata-extract and usnic acid were the most active compounds, especially in anaerobic bacteria. Usnea CO2-extract effectively inhibited the growth of several Gram-positive bacteria like Staphylococcus aureus (including methicillin-resistant strains - MRSA), Propionibacterium acnes and Corynebacterium species. Growth of the dimorphic yeast Malassezia furfur was also inhibited by Usnea-extract. Besides the Usnea-extract, Rosmarinus-, Salvia-, Boswellia- and Harpagophytum-extracts proved to be effective against a panel of bacteria. It is concluded that due to their antimicrobial effects some of the plant extracts may be used for the topical treatment of skin disorders like acne vulgaris and seborrhoic eczema.     

Protective Effect of Enzymatic Extracts from Microalgae Against DNA Damage Induced by H<Subscript>2</Subscript>O<Subscript>2</Subscript>

The enzymatic extracts from seven species of microalgae (Pediastrum duplex, Dactylococcopsis fascicularis, Halochlorococcum porphyrae, Oltmannsiellopsis unicellularis, Achnanthes longipes, Navicula sp. and Amphora coffeaeformis) collected from three habitats (freshwater, tidal pool, and coastal benthic) at Jeju Island in Korea were investigated for their antioxidant activity. Of the extracts tested, the AMG 300 L (an exo 1, 4-α-d-glucosidase) extract of P. duplex, the Viscozyme extract of Navicula sp., and the Celluclast extract of A. longipes provided the most potential as antioxidants. Meanwhile, the Termamyl extract of P. duplex in an H₂O₂ scavenging assay exhibited an approximate 60% scavenging effect. In this study, we report that the DNA damage inhibitory effects of P. duplex (Termamyl extract) and D. fascicularis (Kojizyme extract) were nearly 80% and 69% respectively at a concentration of 100 μg/ml. Thus, it is suggested that the microalgae tested in this study yield promising DNA damage inhibitory properties on mouse lymphoma L 5178 cells that are treated with H₂O₂. Therefore, microalgae such as P. duplex may be an excellent source of naturally occurring antioxidant compounds with potent DNA damage inhibition potential.     

Specific ion effects on macromolecular interactions in Escherichia coli extracts

Protein characterization in situ remains a major challenge for protein science. Here, the interactions of ΔTat‐GB1 in Escherichia coli cell extracts were investigated by NMR spectroscopy and size exclusion chromatography (SEC). ΔTat‐GB1 was found to participate in high molecular weight complexes that remain intact at physiologically‐relevant ionic strength. This observation helps to explain why ΔTat‐GB1 was not detected by in‐cell NMR spectroscopy. Extracts pre‐treated with RNase A had a different SEC elution profile indicating that ΔTat‐GB1 predominantly interacted with RNA. The roles of biological and laboratory ions in mediating macromolecular interactions were studied. Interestingly, the interactions of ΔTat‐GB1 could be disrupted by biologically‐relevant multivalent ions. The most effective shielding of interactions occurred in Mg²⁺‐containing buffers. Moreover, a combination of RNA digestion and Mg²⁺ greatly enhanced the NMR detection of ΔTat‐GB1 in cell extracts.     

褛裳夜蛾的交配行为及雄蛾对性腺提取物的反应节律

详细观察了褛裳夜蛾成虫的交配行为,利用风洞、触角电位技术研究了雄蛾对性腺提取物的反应节律,通过林间诱蛾试验进行了验证,旨在为褛裳夜蛾性信息素的精确提取及性信息素组份分离、鉴定提供依据。研究结果表明:成虫的求偶、交配行为均发生在暗期,有一定的节律性:雌蛾在羽化3d以后开始求偶,1周左右表现最为强烈,3-4日龄雌蛾在暗期4-6h开始求偶,最大求偶率在暗期6-8h;5-9日龄在进入暗期就开始求偶,最大求偶率出现在暗期4-6h。风洞试验表明,3-7日龄的雄蛾对性腺提取物均有明显的性行为反应,5日龄雄蛾在暗期4-6h对性腺提取物的反应最为强烈。3-6日龄的成虫在暗期4-6h开始交配,而7-8日龄的成虫于暗期2-4h开始交配,6日龄的成虫交配率最高,交配高峰在暗期4-8h。雄蛾对性腺提取物的触角电位反应也有一定的节律性:雄蛾对4日龄雌蛾性腺提取物开始有电生理反应,对6-7日龄暗期5h性腺提取物反应最为强烈。林间诱蛾试验测定了性腺提取物的引诱活性,7日龄雌蛾性腺提取物林间诱蛾量最高,引诱高峰在暗期4-6h,该结果也验证了褛裳夜蛾雄蛾对性腺提取物的反应节律。     

Determination of carbohydrates in medicinal plants--comparison between TLC, mf-MELDI-MS and GC-MS

Introduction – Quality control in the pharmaceutical and phytopharmaceutical industries requires fast and reliable methods for the analysis of raw materials and final products. Objective – This study evaluates different analytical approaches in order to recognise the most suitable technique for the analysis of carbohydrates in herbal drug preparations. Methodology – The specific focus of the study is on thin‐layer chromatography (TLC), gas chromatography (GC), and a newly developed mass spectrometric method, i.e. matrix free material enhanced laser desorption/ionisation time of flight mass spectrometry (mf‐MELDI‐MS). Samples employed in the study were standards and microwave‐assisted water extracts from Quercus. Results – TLC analysis proved the presence of mono‐, di‐ and trisaccharides within the biological sample and hinted at the existence of an unknown carbohydrate of higher oligomerisation degree. After evaluation of different derivatisation techniques, GC‐MS confirmed data obtained via TLC for mono‐ to trisaccharides, delivering additionally quantified values under a considerable amount of time. A carbohydrate of higher oligomerisation degree could not be found. The application of mf‐MELDI‐MS further confirmed the presence of carbohydrates up to trisaccharides, also hinting at the presence of a form of tetrasaccharide. Besides this information, mf‐MELDI‐MS delivered further data about other substances present in the extract. Quantitative determination resulted in 1.750, 1.736 and 0.336 mg/mL for glucose, sucrose and raffinose respectively. Conclusion – Evaluation of all three techniques employed, clearly proved the heightened performance of mf‐MELDI‐MS for the qualitative analysis of complex mixtures, as targets do not need modification and analysis requires only a few minutes. In addition, GC‐MS is suitable for quantitative analysis. Copyright © 2011 John Wiley & Sons, Ltd.     

中华补血草多酚提取物对自由基的清除能力

采用超声波辅助浸提—大孔树脂吸附法从中华补血草[ Limonium sinense (Girard) Kuntze]根、根茎、叶和花中获得多酚提取物,并对它们的得率和组成成分进行了初步分析;在此基础上,比较研究了不同部位多酚提取物对DPPH·、·OH和O2的清除能力.实验数据表明:中华补血草根、根茎、叶和花多酚提取物的得率分别为12.42％、5.98％、5.27％和3.98％,差异明显;多酚提取物中总酚、原花色素、黄烷醇和总黄酮含量变化幅度较大,分别为51.87％ ～61.60％、5.62％～39.47％、3.69％～12.46％和2.53％ ～35.97％;其中,总酚、原花色素和黄烷醇含量均以根多酚提取物最高,总黄酮含量以花多酚提取物最高,均与其他部位多酚提取物有显著差异.随质量浓度提高,4个部位多酚提取物对3种自由基的清除率总体上逐渐增大;其中根多酚提取物对DPPH·的清除作用最强,半数清除质量浓度(ρSC50)为38.52 μg·L-1,显著低于阳性对照芦丁和BHT(ρSC50分别为67.40和74.25 μg·L-1);花多酚提取物对·OH和O2的清除能力最强,ρSC50分别为53.51和74.00 μg·L-1,均低于芦丁;总体上,4个部位多酚提取物对DPPH·的清除能力由强至弱依次为根、根茎、叶、花,对·OH和O2的清除能力由强至弱依次为花、根、叶、根茎.结果显示:中华补血草不同部位多酚提取物均具有较强的自由基清除能力,其中,根和花多酚提取物中高含量的原花色素和黄酮类成分分别与其自由基清除能力密切相关.     

Differences in regulation of the first two M-phases in Xenopus laevis embryo cell-free extracts

The first embryonic M-phase is special, being the time when paternal and maternal chromosomes mix together for the first time. Reports from a variety of species suggest that the regulation of first M-phase has many particularities; however, no systematic comparative study of the biochemical aspects of first and the following M-phases has been previously undertaken. Here, we ask whether the regulation of the first embryonic M-phase is modified, using Xenopus cell-free extracts. We developed new types of extract specific for the first and the second M-phase obtained either from parthenogenetic or from in vitro fertilized embryos. Analyses of these extracts confirmed that the amplitude of histone H1 kinase activity reflecting CDK1/cyclin B (or MPF for M-phase Promoting Factor) activity is higher and persists longer than during the second M-phase, and that levels of cyclins B1 and B2 are correspondingly higher during the first than the second embryonic M-phase. Inhibition of protein synthesis shortly before M-phase entry reduced mitotic histone H1 kinase amplitude, shortened the period of mitotic phosphorylation of chosen marker proteins, and reduced cyclin B1 and B2 levels, suggesting a role of B-type cyclins in regulating the duration of mitotic events. Moreover, addition of exogenous cyclin B to the extract prior the second mitosis brought forward the activation of mitotic histone H1 kinase but prolonged the duration of this activity. We also confirmed that the inhibitory phosphorylation of CDK1 on tyrosine 15 oscillates between the first two embryonic M-phases, but is clearly more pronounced before the first than the second mitosis, while the MAP kinase ERK2 tended to show greater activation during the first embryonic M-phase but with a similar duration of activation. We conclude that discrete differences exist between the first two M-phases in Xenopus embryo and that higher CDK1/cyclin B activity and B-type cyclin levels could account for the different characteristics of these M-phases.