固相微萃取结合GC/MS联用技术研究野松茸干品的挥发性成分

采用顶空固相微萃取结合气相色谱-质谱联用技术分析鉴定了野松茸干品中的挥发性风味成分.鉴定出48种风味化合物,占挥发性成分总量的91.44％,其中酸类9种、酯类6种、含氮杂环化合物11种、醇类3种、醛类5种、酮类10种、其他类4种.野松茸干品中主要的芳香成分是桂酸甲酯(42.99％)、3-甲基丁酸(8.56％)、2,6-二甲基吡嗪(8.36％)、2,5-二甲基吡嗪(2.46％)和1-辛烯-3-醇(2.06％).     

芸香甙热裂解研究

在 5 0 0℃和 770℃两种温度下 ,采用热解直接进样的方式 ,研究了烟草中主要的多酚类化合物———芸香甙的热裂解产物。分别鉴定出了 2 4个和 2 3个裂解化合物。通过比较和分析这两种温度下的裂解产物发现 ,芸香甙可以裂解产生大量具有香味的产物。不同温度下的热裂解产物中 ,其中有 16个热裂解产物是相同的 ,但它们的相对含量却不一样。     

固相微萃取-气质法测定土壤挥发性抑菌物质*

运用固相微萃取 气相色谱 质谱法 (SPME GC MS) ,测定了参与土壤抑真菌作用的土壤挥发性成分和土壤细菌挥发性代谢物。通过比较土壤来源和土壤细菌来源的挥发性抑菌成分 ,发现在强挥发性抑菌土壤和土壤细菌代谢物中普遍存在着三甲胺、二甲基二硫醚、3 甲基 2 戊酮、甲基吡嗪、 2 ,5 二甲基吡嗪、N ,N 二甲基辛胺、十九烷等化合物。这些化合物很有可能就是参与土壤抑菌作用 ,特别是挥发性物质抑菌作用的主要成分。另外 ,为深入了解土壤中参与抑菌作用的挥发性化合物提供了简便有效的方法。     

固相微萃取-气质法测定土壤挥发性抑菌物质

运用固相微萃取．气相色谱,质谱法(SPME-GC／MS),测定了参与土壤抑真菌作用的土壤挥发性成分和土壤细菌挥发性代谢物。通过比较土壤来源和土壤细菌来源的挥发性抑菌成分,发现在强挥发性抑菌土壤和土壤细菌代谢物中普遍存在着三甲胺、二甲基二硫醚、3-甲基-2-戊酮、甲基吡嗪、2,5-二甲基吡嗪、N,N-二甲基辛胺、十九烷等化合物。这些化合物很有可能就是参与土壤抑菌作用,特别是挥发性物质抑菌作用的主要成分。另外,为深入了解土壤中参与抑菌作用的挥发性化合物提供了简便有效的方法。     

粗叶悬钩子叶中多酚类与三萜类化合物的研究

采用高效液相色谱-串联质谱（HPLC-MS/MS）联用技术,对中国南方民间用于治疗肝损伤和癌症的药材粗叶悬钩子（Rubus aleaefolius Poir.）叶中的多酚类和三萜类成分进行了分析。通过与文献数据和部分对照品的紫外光谱、负离子模式下的一级和二级质谱信息进行比较,从粗叶悬钩子叶中共分析了18个化合物,鉴定了其中2个多酚类化合物,推测了9个多酚类和7个三萜类化合物的可能结构,并对三萜类化合物的质谱裂解途径进行了解析。基于对三萜类化合物裂解规律的研究,推测了1个三萜,即3-对香豆酰氧基-19α-羟基熊果酸。芦丁、绿原酸和3-对香豆酰氧基-19α-羟基熊果酸在本植物中属首次报道。     

不同提取方法对黔产水香薷挥发性成分的影响

比较不同提取方法对黔产水香薷挥发性成分影响。采用固相微萃取法、水蒸气蒸馏法提取水香薷挥发性成分,用GC-MS联用仪进行测定,结合NIST05和WILEY275谱库鉴定各化合物,峰面积归一化法测定各成分相对含量。利用两种提取方法供从水香薷中鉴别出67个化合物。从固相微萃取法提取物中检测并鉴定了45个化合物,而从水蒸气蒸馏法提取物种检测并鉴定了55个化合物。其中挥发性相同的化合物有33种,两种不同提取方法得到的挥发性成分虽有差异,但主要成分变化不大。体外抗菌试验表明,水香薷挥发性成分具有抑菌作用。这些研究结果为香料植物水香薷的开发提供了科学依据。     

白马骨挥发性化学成分研究

采用水蒸气蒸馏法从白马骨中提取其挥发性成分,再通过气-质联用(GC-MS)技术对所提取的挥发性成分进行分离鉴定,共测定了其中的43种成分.已成功鉴定的成分占样品总量的88.87%,其主要成分为脂肪酸,占挥发性成分总量的52.12%,其次为烯烃化合物12.66%,醇类化合物8.97%,酮类化合物5.90%,酯类化合物3.84%,烷烃化合物3.28%,醛类化合物2.10%.     

Effects of different extraction methods on volatile chemical constituents of the Elscholtzia kachinensis distributed in Guizhou Province

比较不同提取方法对黔产水香薷挥发性成分影响.采用固相微萃取法、水蒸气蒸馏法提取水香薷挥发性成分,用GC-MS联用仪进行测定,结合NIST05和WILEY275谱库鉴定各化合物,峰面积归一化法测定各成分相对含量.利用两种提取方法供从水香薷中鉴别出67个化合物.从固相微萃取法提取物中检测并鉴定了45个化合物,而从水蒸气蒸馏法提取物种检测并鉴定了55个化合物.其中挥发性相同的化合物有33种,两种不同提取方法得到的挥发性成分虽有差异,但主要成分变化不大.体外抗菌试验表明,水香薷挥发性成分具有抑菌作用.这些研究结果为香料植物水香薷的开发提供了科学依据.     

花生中香味成分的研究与分析

笔者通过分析花生中香味成分化合物来对花生中香味成分进行研究。采用的方法为固相微萃取法,此法可以萃取出烘烤花生中的各种风味物质,根据实验我们可以看出:烘烤花生中挥发性的化合物中FD因子的最大值为256,并且FD因子排在前三位(FD因子>64)的在实验中检测出有10种已知的化合物和3种未知的化合物,实验结果显示:已知的化合物分别为:3-甲基丁醛(巧克力味)、2-甲基丁醛(焦糖味)、2-(2-羟乙基)-3-甲基-4-噻唑(鱼腥味)、2,5-二甲基吡嗪(烘烤坚果味)、2,6-二甲基吡嗪(烘烤坚果味)、乙基吡嗪(爆米花味)、2-乙基-5-甲基吡嗪(草味,坚果味)、呋喃醛(生土豆味)、3-乙基-2,5-二甲基吡嗪(咖啡味)、苯乙醛(甜芳香味)等。通过本次的实验,可以得出结论:2,5-二甲基吡嗪、乙基吡嗪等吡嗪类和醛类化合物是花生中香味的主要成分,对花生散发香味的贡献最大,酮类化合物对花生的风味贡献率较醛类来讲较低。     

贵州产辣蓼挥发性成分分析

采用固相微萃取法和水蒸气蒸馏法提取出贵州产辣蓼的挥发性成分,运用GC-MS联用技术对挥发性成分进行分析鉴定,并采用峰面积归一法确定各成分的相对含量。固相微萃取物中鉴定出48个化合物,主要有β-石竹烯(27.02%),正癸醇(14.39%),十二醛(12.96%)等,水蒸气蒸馏物中鉴定出27个化合物,β-红没药烯(19.00%),补身树醇(15.25%),十二醛(14.41%)等。辣蓼中挥发性成分经两种不同提取方法成分存在明显差异。与文献数据比较,不同省区产的辣蓼挥发性成分差异很大。体外抗菌试验表明,辣蓼挥发性成分具有抑菌作用,这些研究结果为辣蓼的有效利用提供了依据。     

Chemical studies of the antioxidant mechanism of tea catechins: radical reaction products of epicatechin with peroxyl radicals

Tea catechins, an important class of polyphenols, have been shown to have antioxidant activity and are thought to act as antioxidants in biological systems. However, the mechanisms of their antioxidant reactions remain unclear. The objective of this study was to characterize the reaction products of epicatechin with peroxyl radicals generated by thermolysis of the azo initiator azo-bisisobutyrylnitrile (AIBN). Structural elucidation of these products can provide insights into specific mechanisms of antioxidant reactions. Eight reaction products were isolated and identified using high-field 1D and 2D NMR spectral analysis. The observation of these compounds confirmed that the B-ring is the initial site for formation of reaction products in the peroxyl radical oxidant system.     

Hydrocarbons and fatty acid methyl esters in bacterial biomass before and after physicochemical treatment

Hydrocarbons and fatty acid methyl esters were identified by chromatography-mass spectrometry in extracts of the native biomass of bacteria: chemoorganoheterotrophic Arthrobacter sp. and Pseudomonas aeruginosa and chemolithoautotrophic Carboxydothermus sp. Ultrasound treatment of bacterial biomass and mild thermolysis were shown to promote formation of a broad spectrum of hydrocarbons from bacterial biomass. The biomarker stigmastane belonging to the sterane group was found in P. aeruginosa biomass after thermolysis at 110°C in an open vial. Alkane composition in P. aeruginosa biomass before and after thermolysis at 300°C in a sealed container remained unchanged, indicating the possibility of preservation of hydrocarbons of bacterial origin in sealed layers under high temperature and elevated pressure.     

A new study of cell disruption to release recombinant thermostable enzyme from Escherichia coli by thermolysis

Extraction of intracellular protein from Escherichia coli is traditionally achieved by mechanical, chemical or enzymatic disruption technology. In this study, a novel thermolysis method was used to disrupt E. coli cells to release a recombinant thermostable esterase. We found that heat treatment of E. coli was highly effective to destroy the integrity of bacterial cell walls and release the recombinant hyperthermophilic esterase at temperatures above 60 degrees C. The effects of temperature, pH and cell concentration on the efficiency of cell disruption were examined. The most effective temperature for cell disruption was at 80 degrees C. The pH and cell concentration had only minor effect on the release of the hyperthermophilic esterase. In addition, we found that the hyperthermophilic esterase could be purified at the early stage of the thermolysis, which is a major advantage of the thermolysis method. Finally, a comparison between thermolysis and traditional methods for the disruption of cells and the release of the thermostable enzyme was made.     

The formation of chromophores through amino acid thermolysis and their possible role as prebiotic photoreceptors

The thermal polymerization of amino-acid mixtures was studied at various temperatures and reaction times with specific emphasis on the formation of fluorescent chromophores. The reaction conditions appeared to have a pronounced effect on the ratio of synthesized chromophores and biuret-positive material. During thermolysis of equimolar mixtures of lysine, alanine and glycine or lysine, aspartic acid and glycine small amounts of pteridines and flavines are formed, which are often covalently linked to the thermal oligomer. These heterocyclic compounds are likely formed by condensation reactions of the amino acid break-down and conversion products. Reaction schemes that describe the processes are proposed. The significance of these chromoproteinoids is discussed in respect to prebiotic redox reactions and photoinduced processes.     

A mechanistic study of thermal C-H reductive elimination from a six-coordinate d iridium complex

The thermolysis of trans-IrL₂(CO)Cl(H)(C₆H₅) (1abd; L=P(i-Pr)₃; H trans to CO) produces benzene and the Vaska-type complex IrL₂(CO)Cl. A mechanistic study of the reaction has shown that 1a reversibly loses CO at 120 °C (as evidenced by the incorporation of ¹³CO) and isomerizes to the previously unreported 1b (H trans to Cl). It was found that 1b is the complex primarily responsible for the formation of benzene upon thermolysis under CO atmosphere; direct loss of benzene from 1a was determined to be, at most, a minor pathway. Benzaldehyde was also formed as a product of thermolysis of 1a under CO atmosphere. The first-order rate constant for benzene elimination in the absence of CO was found to be 8.5 × 10⁻⁵ s⁻¹. The presence of only 5 Torr CO results in a decrease to 2.0 × 10⁻⁵ s⁻¹, but little further inhibition is observed above 5 Torr CO. Added dihydrogen (100 Torr) was found to effect a novel catalysis of benzene elimination from 1a in the absence of CO atmosphere; it is suggested that trace amounts of dihydrogen, present in solutions of 1a, are responsible for the enhanced rate of elimination in the absence of CO. The thermolysis of 1-d₆ in toluene was found to proceed without any toluene incorporation, implying that arene loss is irreversible.     

Peroxyl Radical Scavenging by a Series of Coumarins

Sixteen plant-derived or synthetic coumarins with various hydroxyl and other substitutions were tested for their ability to scavenge alkylperoxyl radicals generated in the aqueous phase by the controlled thermolysis of 2,2'-azo-bis-(2-amidinopropane) dihydrochloride (ABAP). Protection by coumarins against inactivation of lysozyme by the radicals was assayed by measuring the loss of turbidity of suspensions of M. lysodeikticus. Ten of the coumarins were potent scavengers of aqueous peroxyl radicals with activities comparable to n-propyl gallate, desferrioxamine, ferrioxamine and trolox c, yielding IC50 values in the range 21 to 92 micromolar. The presence of 6,7-ortho-dihydroxy functions gave compounds of the greatest potency. Scavenging activity was unrelated to ability to chelate iron ions. The active coumarins are attractive candidates for evaluation as protective agents against disorders in which oxidative stress is implicated.     

Cell thermolysis – A simple and fast approach for isolation of bacterial laccases with potential to decolorize industrial dyes

Laccases belong to multicopper oxidases and are widespread in nature. Currently, mainly fungal laccases are applied in biotechnological processes. One reason for this is that fungal laccases are much better studied. Compared to fungal laccases, bacterial laccases possess some advantageous characteristics like high stability at elevated temperatures and alkaline pH values. Intracellular recombinant expression of bacterial laccases in E. coli makes however downstream processing more complex and time-consuming compared to extracellular expression of fungal enzymes. Here, we demonstrate that cell disruption by cell thermolysis is an efficient and simple method for the isolation and partial purification of recombinant bacterial laccases. Three different laccases, Tth from Thermus thermophilus, CotA from Bacillus subtilis and Ssl1 from Streptomyces sviceus, were used to compare cell disruption by cell thermolysis with sonication and high-pressure homogenization, with and without subsequent heat treatment. Cell thermolysis resulted in high laccase activities per gram of cell wet weight and in the highest specific activities of the laccases. For example, specific activity of Tth laccase after cell thermolysis was 469-fold higher than after sonication. Furthermore, high decolorization activity towards indigo carmine and alizarin red S of these laccases, isolated via cell thermolysis, demonstrate their potential for technical applications.     

EPR Spin Trapping Study of the Decomposition of Azo Compounds in Aqueous Solutions by Ultrasound: Potential for Use as Sonodynamic Sensitizers for Cell Killing

Sonodynamic therapy, a promising new approach to cancer treatment, is based on synergistic cell killing by combination of certain drugs (sonosensitizers) and ultrasound. Although the mechanism of sonodynamic action is not understood, the role of free radicals produced from sonosensitizers by ultrasound is implicated. In this work, we studied formation of free radicals during the decomposition of several water-soluble azo compounds by 50 kHz ultrasound in aqueous solutions. Using the spin trap 3, 5-dibromo-4-nitrosobenzene sulfonate (DBNBS) tertiary carbon-centered radicals from 2, 2'-azobis (N,N'-dimethyl-eneisobutyramidine) dihydrochloride (VA-044), 2-(carbamoylazo)-isobutyronitrile (V-30), and 2, 2'-azobis (2-amidinopropane) dihydrochloride (AAPH) and CH3 radicals from 1, 1'-azobis (N,N'-dimethylformamide) (ADMF) were detected in argonsaturated solutions and the corresponding oxygen-centered radicals (alkoxyl and peroxyl) from VA-044, V-30, and AAPH were identified using the spin trap 5, 5'-dimethyl-l-pyrroline-N-oxide (DMPO) in aerated sonicated solutions. No free radicals from 4, 4'-dihydroxyazobenzene-3, 3'-dicarboxylic acid, disodium salt (DHAB) could be found in either system. While VA-044 and AAPH could also be readily decomposed by heat (42.5°C and 80°C), V-30 decomposition only occurred in the ultrasound-exposed solutions. The most likely mechanism of decomposition of azo compounds by ultrasound is their thermolysis in the heated shell of the liquid surrounding ca vita ting bubbles driven by ultrasound and/or by pyrolysis inside these bubbles. Experiments using scavengers of ·OH and ·H, which are produced by sonolysis in aqueous solutions, demonstrated that these radicals are not involved in the ultrasound-mediated radical production from the azo compounds. Due to the known cytotoxic potential of free radicals produced from azo compounds, the use of these compounds as ultrasound sensitizers appears to be a promising approach for sonodynamic cell killing.     

A simple method of genomic DNA extraction suitable for analysis of bulk fungal strains

Aims: A simple and rapid method (designated thermolysis) for extracting genomic DNA from bulk fungal strains was described. Methods and Results: In the thermolysis method, a few mycelia or yeast cells were first rinsed with pure water to remove potential PCR inhibitors and then incubated in a lysis buffer at 85°C to break down cell walls and membranes. This method was used to extract genomic DNA from large numbers of fungal strains (more than 92 species, 35 genera of three phyla) isolated from different sections of natural Ophiocordyceps sinensis specimens. Regions of interest from high as well as single‐copy number genes were successfully amplified from the extracted DNA samples. The DNA samples obtained by this method can be stored at ?20°C for over 1 year. Conclusions: The method was effective, easy and fast and allowed batch DNA extraction from multiple fungal isolates. Significance and Impact of Study: Use of the thermolysis method will allow researchers to obtain DNA from fungi quickly for use in molecular assays. This method requires only minute quantities of starting material and is suitable for diverse fungal species.