Assessment of bog-body tissue preservation by pyrolysis-gas chromatography/mass spectrometry

Flash pyrolysis-gas chromatography/mass spectrometry (py-GC/MS) was used to assess the quality and mechanism of protein preservation in the tissue of Iron Age bog bodies from Lindow, UK, and south-eastern Drenthe, The Netherlands. Abundant pyrolysis products of the fresh skin tissue, including 2,5-diketopiperazines of Pro-Gly, Pro-Ala, Pro-Val, Pro-Pro and Hyp, were readily assigned to specific amino acid or dipeptide moieties. Comparison of the pyrolysates of the bog-body tissues with that of modern samples revealed qualitative similarities suggesting good preservation of the collagen and non-collagenous proteins in the ancient tissues. Examination of the pyrolysates of samples of fresh calf skin, which had been treated with various vegetable tanning agents, clearly revealed markers of non-hydrolysable tannins including 1,2-benzenediol, 1,3-benzenediol and 1,2,3-benzenetriol, although chromatographic quality inevitably diminished with increasing functionalization of the compounds. Such markers were not detected in the pyrolysates of the bog-body tissues. Instead 4-isopropenylphenol, a characteristic pyrolysis product of Sphagnum moss, was detected in both solvent-extracted and base-treated samples of tissue. The presence of 4-isopropenylphenol in the pyrolysates of the bog-body tissues provides evidence that their preservation involves reactions of amino acids with sphagnum acid, and possibly other agents derived from the peat. The study constitutes the first chemical characterization of the pyrolysis products of modern and ancient collagen.     

承钢高炉喷吹用煤的等离子高温热解性能

利用电弧等离子体高温热解装置、多功能气相色谱仪和扫描电镜(SEM)对承钢高炉喷吹煤种进行高温热解实验,研究了配煤和添加助燃剂后煤粉的热解率、热解气体产率、热解气体组成的变化规律。结果表明,配煤和添加助燃剂都能提高煤粉的热解率和气相产物的产率,当烟煤配比在60%(质量分数)时,配煤方案的热解率最高为55.17%,添加助燃剂能够继续强化配煤的热解性能。热解气体中CO和H_2的含量随烟煤配比和助燃剂增加而增加,小分子烃类物质的产量减少,残渣颗粒表面出现大量不规则形状的突起,并伴有较多的裂纹、空隙和大量的孔状结构,进而提高煤粉在高炉风口区域的高温热解性能。     

Separation and characterization of histones and acidic proteins from heart muscle and calf thymus

A method for the isolation of histones and acidic proteins extracted from rabbit heart and calf thymus chromatins has been studied. The histones present five major electrophoretic fractions in both tissues. Only lysine rich fraction in the heart splits into three peptides of different mobility. Amino acid composition of total histones shows values very similar in almost all components. The acidic proteins of rabbit heart and calf thymus have been characterized by sodium dodecyl sulphate polyacrylamide gel electrophoresis and by amino acid composition. The electrophoretic pattern reveals reproducible heterogeneous banding pattern from the two sources with different degree of phosphorylation. The amino acid composition of total acidic proteins is quite similar with the exception of serine content that in the heart is most elevated.     

An improved chemical approach toward the C-terminal sequence analysis of proteins containing all natural amino acids

An improved chemical method, capable of derivatizing all natural amino acids to their corresponding thiohydantoins, is described. This involves activation by acetyl chloride in TFA followed by derivatization with ammonium thiocyanate. Possible interference of reactive side chains was investigated by reacting N-acetylamino acids as well as several peptides with propionyl chloride instead of acetyl chloride. The products were characterized by PDMS mass spectrometry and 1H-NMR. This chemical method allows, for the first time, complete derivatization of N-acetylproline to proline thiohydantoin. Applying this chemistry to peptides with a C-terminal proline, the yields for formation of proline thiohydantoin were found to be up to 60%, depending on the peptide sequence. The previous inability to derivatize C-terminal proline to thiohydantoin was thought to stem from the fact that proline cannot form the oxazolonium ion required for efficient reaction with the thiocyanate ion. However, we have found mass spectrometric evidence for the existence of a proline oxazolonium ion, under basic as well as under acidic conditions. This improvement in derivatization of C-terminal amino acids including proline is a major step forward in the development of a general chemical C-terminal sequencing method that permits the C-terminal sequence analysis of proteins of any amino acid composition.     

微波热解法制备氧化铈过程的可视化研究

针对传统液相法制备氧化铈纳米颗粒存在工艺流程复杂、高污水排放等问题,提出了一种高效绿色的实验方案,以七水合氯化铈为原料,采用微波射流热解技术制备出了高纯度氧化铈纳米颗粒.通过X射线衍射仪(XRD)、扫描电镜(SEM)和能谱仪(EDS)分析手段对产物进行了表征,借助数值模拟手段可视化分析了各物理场、各组分分布.考察了不同工艺条件(热解温度、气相速度、和添加柠檬酸)对实验产物中残余氯根含量和产物微观形貌的影响.结果表明,热解温度达到500℃时便可获得单相氧化铈,温度越高氧化铈纯度越高,颗粒形貌越规则.增大气相入口速度导致产物残余氯根增多,但有利于改善颗粒团聚.添加柠檬后氧化铈从球状颗粒逐渐破碎为伴有少量多孔结构的不规则形状颗粒,颗粒比表面积增大.柠檬酸浓度大于0.1 mol?L^-1后利于减少氯根含量.     

天然气还原热解硫酸镁制备高纯氧化镁

利用盐湖氯化镁和硫酸制备七水硫酸镁,将七水硫酸镁脱水得到无水硫酸镁.采用天然气还原热解无水硫酸镁得到高纯氧化镁.通过单因素实验考察了还原热解温度、热解时间、硫酸镁的粒径和天然气气体流量对硫酸镁转化率的影响,通过正交试验优化了还原热解的条件.采用X射线衍射和扫描电镜对还原热解产物进行分析和表征.氧化镁的最佳制备条件:热解温度为1000℃,热解时间为30min,硫酸镁粒径为75μm,天然气气体流量为25mL·min-1.温度是影响硫酸镁转化率的主要因素.在最佳制备条件下,硫酸镁的转化率达到99.27%,氧化镁的纯度达到99.5%,制取的氧化镁单分子均匀,表面为多孔蓬松,具有高比表面积.      

Determination of amino acids in biological fluids by capillary gas chromatography with nitrogen-phosphorus selective detection

A selective and sensitive method for the determination of protein and non-protein amino acids in biological fluids by capillary gas chromatography (GC) has been developed. The amino acids in the samples were directly converted into their N(O,S)-isobutoxycarbonyl methyl ester derivatives and measured by GC with nitrogen-phosphorus selective detection (NPD) using a DB-17ht capillary column. Using this method, the derivatives of the 21 protein amino acids and the 25 non-protein amino acids provided excellent NPD responses and were quantitatively and reproducibly resolved within 28 min. The lower detection limits of these amino acids, at a signal-to-noise ratio of 3, were ca. 6-150 pg injected. The calibration curves for each amino acid in the range of 0.02-2 micrograms were linear and sufficiently reproducible for quantitative analysis. This method was successfully applied to small urine and serum samples without prior clean-up; there was no evidence of interference from coexisting substances. Overall recoveries of amino acids added to urine and serum samples were 83-112%. The intra-assay and inter-assay R.S.D. of amino acids in these samples were 0.3-8.9% (n = 3) and 1.9-15.8% (n = 3), respectively.     

低阶煤热解与兰炭生产工艺研究进展

当今社会,化石能源日益枯竭,充分高效利用现有资源和开发新的可再生资源十分必要.低阶煤挥发分高,燃烧性和反应性较好,直接燃烧无法充分发掘其所蕴含的潜在价值.热解技术作为一种能够高效利用煤炭资源的技术,受到许多学者的重视.根据低阶煤在不同转化阶段反应性不同的特点,通过控制煤的热解过程可以制得兰炭.兰炭的气化活性优于焦炭和无...     

Identification of the conserved coding sequences of three chitin synthase genes in Fonsecaea pedrosoi

Primers having designs based on highly conserved stretches in the deduced amino acid sequences of chitin synthase (CHS) genes were used in PCR reactions to amplify 600 bp and 366 bp products from the genomic DNA of three major causal agents of chromoblastomycosis. Cloning and sequencing of the PCR products of one of these fungi, Fonsecaea pedrosoi, identified three CHS sequences designated as FpCHS1, FpCHS2 and FpCHS3. FpCHS1 and FpCHS2 were homologous to regions of CHS1 and CHS2 of Saccharomyces cerevisiae, and their derived amino acid sequences fell into chitin synthase classes I and II, respectively. FpCHS3 was homologous to a region of the CAL1/CSD2 gene of S. cerevisiae, which codes for the chitin synthase three (Chs3) enzyme in that fungus. Phylogenetic trees constructed using the deduced amino acid sequences of PCR-amplified CHS products from many fungi clustered F. pedrosoi with other dematiaceous fungi, providing new molecular evidence for the genetic relatedness of these organisms. The identification of these CHS genes in F. pedrosoi will facilitate future studies of the functional roles of chitin synthases in the unique in vivo dimorphism exhibited by chromoblastomycotic fungi.     

New cyclic peptides from the ascidian Lissoclinum patella

Four new cyclic peptides, patellamide G (2) and ulithiacyclamides E-G (3-5), along with the known patellamides A-C (6-8) and ulithiacyclamide B (9), were isolated from the ascidian Lissoclinum patella collected in Pohnpei, Federated States of Micronesia. The planar structures of these peptides were determined from 1D and 2D 1H and 13C NMR spectra. The absolute stereochemistries of the amino acid units, except for cysteine, were assigned by chiral GC analysis of N(O)-trifluoroacetyl isopropyl ester derivatives of amino acids obtained by acid hydrolysis of the intact and ozonized peptides. The structures of ulithiacyclamides E-G (3-5) were confirmed by chemical conversion. Patellamides B (7) and C (8) exhibited in vitro modulation of multidrug resistance in CEM/VBL100 cells.     

Two-dimensional map of Haemophilus influenzae following protein enrichment by heparin chromatography

Two-dimensional gel electrophoresis separates several hundred protein molecules in one single experiment and is efficiently used to study the products expressed by different genomes. Low-copy-number gene products are invisible on a stained two-dimensional map and must be enriched such that sufficient amounts are present for visualization and identification. We investigated the enrichment of proteins of the bacterium Haemophilus influenzae by chromatography on immobilized heparin which has affinity for growth and protein biosynthesis factors. Total soluble proteins of the microorganism were fractionated on Heparin-Actigel which resulted in enrichment of approximately 160 proteins. The eluates, representing about 40% of the applied proteins, were analyzed by two-dimensional gel electrophoresis and the protein spots were characterized by amino acid composition analysis and matrix-assisted laser desorption ionization mass spectrometry. The proteins enriched by chromatography on the heparin gel were not exclusively low-copy-number gene products and they did not exclusively belong to one single class of proteins. The proteins that bound to the heparin gel are indicated in a two-dimensional protein map which includes more than 110 newly identified proteins.     

Detection of amphetamine and methamphetamine in urine by gas chromatography/Fourier transform infrared (GC/FTIR) spectroscopy

New methods for identification of amphetamines have been employed using gas chromatography/Fourier transform infrared spectroscopy (GC/FTIR). These methods have provided identification of the drug and its metabolites, with detection at the low picogram levels or less than 10 ng/mL. Developments in cryogenic sample deposition for GC/FTIR spectroscopy have increased the sensitivity of GC/FTIR to levels that match or surpass that of gas chromatography/mass spectrometry (GC/MS). This advancement in technology has allowed the highly selective ability of infrared spectroscopy to be used for identification and quantitation in studies where the analytes are in low concentrations. The limits of detection (LOD), quantitation (LOQ), and linearity (LOL), and precision have been determined in this study, and these instrumental parameters have been compared with those of established techniques.     

Liquid chromatography/tandem mass spectrometry of synthesis products associated with the viral protein US11

US11 is a small basic protein composed of 161 amino acid residues, and is among the most abundant viral proteins in cells infected with herpes simplex viruses HSV1 and HSV2. The amino acid sequence [91-121] is considered essential for the binding of this protein with RNA. Automated solid phase synthesis of this fragment resulted in a crude reaction mixture containing the desired sequence as well as a number of unknown side products. On-line liquid chromatography/electrospray mass spectrometry (LC/ES-MS) and LC/ES tandem mass spectrometry (MS/MS) allowed the identification of the separated components and furnished relevant sequencing information. The unusual sequences of the monitored components, which consist of a tandemly repeated three-amino-acid motif with the sequence Arg-X-Pro, where X is an acidic or uncharged polar amino acid residue, yielded product ion spectra lacking substantial sequence information and rich in fragment ions manifesting the neutral losses 17, 42 and 60 u.     

Effects of non-REM sleep on ventilation and respiratory mechanics in adults with cystic fibrosis

A polysaccharide containing 2-acetamido-2-deoxy-D-glucose (GlcNAc), 2-acetamido-2-deoxy-L-fucose (FucNAc), and 2-acetamido-2-deoxy-D-galacturonic acid (GalNAcA) was isolated from an aqueous phenol extract of lipid-free, isolated cell walls of the reference strain for Acinetobacter baumannii serogroup O5, by mild acid hydrolysis of the extract and chromatography of the water-soluble products on Sephadex G-50. By means of NMR studies, methylation analysis, carboxyl reduction and chemical degradations, the repeating unit of the polymer was identified as a branched tetrasaccharide of the structure shown. The serologically active polymer is believed to correspond to the side chain of the O5 lipopolysaccharide: [table: see text]     

Semisynthesis of cytotoxic proteins using a modified protein splicing element

Two cytotoxic proteins, bovine pancreatic ribonuclease A (RNase A), and a restriction endonuclease from Haemophilus parainfluenzae (HpaI), were produced using a novel semisynthetic approach that utilizes a protein splicing element, an intein, to generate a reactive thioester at the C-terminus of a recombinant protein. Nucleophilic attack on this thioester by the N-terminal cysteine of a synthetic peptide ultimately leads to the ligation of the two reactants through a native peptide bond. This strategy was used to produce RNase A and HpaI by isolating inactive truncated forms of these proteins, the first 109 and 223 amino acids of RNase A and HpaI, respectively, as fusion proteins consisting of the target protein, an intein, and a chitin binding domain. Thiol-induced cleavage of the precursor led to the liberation of the target protein with a C-terminal thioester-tag. Addition of synthetic peptides representing the amino acids missing from the truncated forms led to the generation of full-length products that displayed catalytic activity indicative of the wild-type enzymes. The turnover numbers and Km for ligated and renatured RNase A were 8.2 s(-1) and 1.5 mM, in good agreement with reported values of 8.3 s(-1) and 1.2 mM (Hodges & Merrifield, 1975). Ligated HpaI had a specific activity of 0.5-1.5 x 10(6) U/mg, which compared favorably with the expected value of 1-2 x 10(6) U/mg (J. Benner, unpubl. obs.). Besides assisting in the production of cytotoxic proteins, this technique could allow the easy insertion of unnatural amino acids into a protein sequence.     

邯钢高炉喷吹煤粉的快速热解机制

为了提高邯钢高炉喷吹煤比,模拟煤粉在高炉内的热解。以邯钢喷吹用长治煤(以下简称CL)和大湾煤(以下简称DW)为原料,采用等离子体进行快速热解,计算反应后煤粉的分解率,利用气相色谱仪对气体产物进行分析以及用扫描电镜(SEM)观察反应产物的形貌特征。试验结果表明,CL和DW的分解率分别为43.10%和52.04%,气相产物主要为CO、H2、CH4、C2H2及少量C2H4等气体,热解产物的粒径减小,形貌发生明显变化。在CL煤的基础上配加不同比例的DW后,煤粉颗粒出现了孔状结构,因此可以提高炉内风口回旋区固定碳颗粒的燃烧率,为提高煤粉燃烧率提供理论依据。     

Coal pyrolysis in a rotary kiln: Part I. Model of the pyrolysis of a single grain

A mathematical model is presented which describes the pyrolysis of a single grain of coal and is designed to be incorporated into an overall model simulating the rotary kiln coal pyrolysis process. The grain model takes into account the principal physical phenomena occurring during the conversion of coal to coke, namely, heat transfer toward and within the grain, drying of the coal, and the evolution of volatile species. Particular care has been taken in the determination of the thermophysical and kinetic parameters necessary for the model. Thus, the drying kinetics for Lorraine coal were measured by thermogravimetry. The kinetics of pyrolysis were determined by both thermogravimetry and gasphase chromatography, in order to separately monitor the evolution of the nine gaseous species considered. The true specific heat and the thermal conductivity of the solid were also measured as a function of temperature. The numerical model, based on the finite-volume method, calculates the temperature, composition, and mass flow rates for the different gases evolved at each point in the grain at any instant of time. The model was, finally, validated by comparing the calculated and measured values of the overall conversion of the pyrolysis reaction and the temperature at the center of the grain.     

Lubricant pyrolysis during sintering of powder metallurgy compacts

The chemistry surrounding the pyrolysis of N,N′-ethylenebisstearimide (EBS) compacted with iron powder is described for the first time. Heat treatment is carried out in a 5 vol pct hydrogen atmosphere (balance nitrogen) over the 100 °C to 850 °C range. The exhaust from the furnace is monitored by Fourier transform infrared and dispersive ultraviolet absorption spectroscopy; condensable materials are analyzed by gas chromatography/mass spectrometry (GC/MS). A wide range of analytes emitted from the preceding process were characterized. The aliphatic CH stretch in the 3000 to 2700 cm⁻¹ range and the asymmetric CO stretch in gaseous CO₂ at 2350 cm⁻¹ are excellent indicators of the extent of delubrication. A bimodal CO emission phase is observed in the temperature window between delubrication and sintering. Three major large molecule reaction products, along with five minor compounds, are identified by GC/MS. A preliminary reaction mechanism is inferred based on product analysis and known organic chemistry. It appears that hydrolysis of EBS competes with γ-H abstraction yielding an N-vinyl amide and stearamide, which undergoes further reaction. Hydrolysis affords stearic acid, which decarboxylates to heptadecane, and 2-heptadecyl-4,5-dihydroimidazole via ring closure of the corresponding amino-amide. This paper is dedicated to Robert M. Becker. Bob died tragically in an automobile accident in December 2003. He was a good friend and valued colleague who will be greatly missed.     

不同热解工艺下半焦的微观结构特点及其对燃烧性能的影响

摘要：对比研究了3种直立内热炉半焦(熄焦方式分别为水泡熄焦、喷水熄焦及烟气干熄焦)及一种流化床热解半焦的微观结构及燃烧性能。通过对半焦组成、孔隙结构、有机官能团、碳化学结构及半焦微晶结构分析,获得了不同低温热解工艺半焦的组成及微观结构特点,明确了组成及微观结构与燃烧性能之间的关系。结果表明,不同热解工艺半焦的组成和微观结构差异明显,其对半焦的燃烧性能影响显著。干熄焦碳化学结构缺陷比例大,快速热解半焦孔隙结构发0达、挥发分高,与水喷焦相比水泡焦石墨化程度低;水喷焦燃烧性能优于水泡焦,干法所得半焦的燃烧性优于湿法;半焦的挥发分含量决定其燃点,在快升温速率条件下孔隙结构对半焦的燃烧性能影响较大,而慢升温速率条件下有序化度对半焦的燃烧性能影响较大。