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1.
Oligosaccharides were analyzed by a combination of high-performance liquid chromatography (HPLC) and mass spectrometry (MS). First, oligosaccharides labeled with 2-aminopyridine were studied to see if they could be analyzed by MS under the conditions used for separation by HPLC. Pyridylamino (PA)-oligosaccharides could be analyzed under these conditions, although the mass spectra were affected. Then, liquid chromatography-mass spectrometry was used to analyze a PA-oligosaccharide mixture derived from human immunoglobulin G. The PA-oligosaccharides were separated on a reversed-phase column and mass-analyzed directly. The observed molecular weights were close to or identical to those expected from the structures, which were estimated from the elution position on HPLC. This method is rapid and simple, as the mass spectrometer can give the accurate molecular weight of each PA-oligosaccharide in one chromatography run, even if the HPLC separation is incomplete. This method can be used to extend the so-called two-dimensional mapping of PA-oligosaccharides. The structure can be studied in greater detail by tandem MS. 相似文献
2.
Maria Christiernin Anna B Ohlsson Torkel Berglund Gunnar Henriksson 《Plant Physiology and Biochemistry》2005,43(8):777-785
Hybrid aspen (Populus tremula x tremuloides) cell cultures were grown for 7, 14 and 21 days. The cell cultures formed primary cell walls but no secondary cell wall according to carbohydrate analysis and microscopic characterization. The primary walls were lignified, increasingly with age, according to Klason lignin analysis. Presence of lignin in the primary walls, with a higher content in 21-day old cells than in 7-day old cells, was further supported by phloroglucinol/HCl reagent test and confocal microscopy after both immunolocalization and staining with acriflavin. Both laccase and peroxidase activity were found in the cultures and the activity increased during lignin formation. The lignin from the cell culture material was compared to lignin from mature aspen wood, where most of the lignin originates in the secondary cell wall, and which served as our secondary cell wall control. Lignin from the cell walls was isolated and characterized by thioacidolysis followed by gas chromatography and mass spectrometry. The lignin in the cell cultures differed from lignin of mature aspen wood in that it consisted exclusively of guaiacyl units, and had a more condensed structure. Five lignin structures were identified by mass spectrometry in the cell suspension cultures. The results indicate that the hybrid aspen cell culture used in this investigation may be a convenient experimental system for studies of primary cell wall lignin. 相似文献
3.
A thermospray high-performance liquid chromatography/mass spectrometry method for determination of serum dehydroepiandrosterone sulfate is described. The steroid was measured intact using [7,7-2H2]dehydroepiandrosterone sulfate as internal standard. The analysis was carried out in the negative ion mode by determining the peak height ratio of the molecular anions of the analyte and internal standard. The method was used to determine the steroid in serum from 15 male and female normal adults and the following values were obtained: males, 272 +/- 45 micrograms/dl (range, 197 to 331 micrograms/dl) and females, 215 +/- 67 micrograms/dl (range, 107 to 347 micrograms/dl). In addition, dehydroepiandrosterone sulfate was measured by high-performance liquid chromatography/mass spectrometry and radioimmunoassay (a commercial kit) on 25 individuals of all age groups. There was strong correlation between the values obtained, but the radioimmunoassay values were generally double those obtained by high-performance liquid chromatography/mass spectrometry. Three other steroid sulfates, androsterone sulfate, epiandrosterone sulfate, and androst-5-ene-3 beta, 17 beta-diol sulfate, were also assayed. In males, these had mean values of 112, 44, and 13 micrograms/dl and, in females, they had mean values of 84, 25, and 6 micrograms/dl, respectively. Radioimmunoassay cross-reactivity measurement for these steroids (as reference compounds) showed that they were unlikely to contribute greatly to the discrepancy between radioimmunoassay and high-performance liquid chromatography/mass spectrometry values. 相似文献
4.
M Furuya S Akashi K Hirayama 《Biochemical and biophysical research communications》1989,163(2):1100-1106
The primary structure of human epidermal growth factor (hEGF), which was produced by Escherichia coli using recombinant DNA technique, has been studied by tandem mass spectrometry. The molecular weight of hEGF (about 6200 amu) was determined by fast atom bombardment mass spectrometry. Then reduced and carboxymethylated hEGF was digested by chymotrypsin into seven peptides which could cover the whole sequence of hEGF. The amino acid sequences of five of these seven peptides could be confirmed by tandem mass spectrometry with or without isolation by high-performance liquid chromatography (HPLC). After isolation by HPLC, the other two peptides were digested with trypsin or thermolysin into small peptides, and sequenced by tandem mass spectrometry. 相似文献
5.
Developmental Control of Lignification in Stems of Lowland Switchgrass Variety Alamo and the Effects on Saccharification Efficiency 总被引:1,自引:0,他引:1
Hui Shen Chunxiang Fu Xirong Xiao Tui Ray Yuhong Tang Zengyu Wang Fang Chen 《Bioenergy Research》2009,2(4):233-245
The switchgrass variety Alamo has been chosen for genome sequencing, genetic breeding, and genetic engineering by the US Department of Energy Joint Genome Institute (JGI) and the US Department of Energy BioEnergy Science Center. Lignin has been considered as a major obstacle for cellulosic biofuel production from switchgrass biomass. The purpose of this study was to provide baseline information on cell wall development in different parts of developing internodes of tillers of switchgrass cultivar Alamo and evaluate the effect of cell wall properties on biomass saccharification. Cell wall structure, soluble and wall-bound phenolics, and lignin content were analyzed from the top, middle, and bottom parts of internodes at different developmental stages using ultraviolet autofluorescence microscopy, histological staining methods, and high-performance liquid chromatography (HPLC). The examination of different parts of the developing internodes revealed differences in the stem structure during development, in the levels of free and well-bound phenolic compounds and lignin content, and in lignin pathway-related gene expression, indicating that the monolignol biosynthetic pathway in switchgrass is under complex spatial and temporal control. Our data clearly show that there was a strong negative correlation between overall lignin content and biomass saccharification efficiency. The ester-linked p-CA/FA ratio showed a positive correlation with lignin content and a negative correlation with sugar release. Our data provide baseline information to facilitate genetic modification of switchgrass recalcitrance traits for biofuel production. 相似文献
6.
An unpolluted process of wheat straw fractionation by steam explosion coupled with ethanol extraction was studied. The wheat straw was steam exploded for 4.5 min with moisture of 34.01%, a pressure of 1.5 MPa without acid or alkali. Hemicellulose sugars were recovered by water countercurrent extraction and decolored with chelating ion exchange resin D412. The gas chromatography (GC) and high-performance liquid chromatography (HPLC) analysis results indicated that there were organic acids in the hemicellulose sugars and the ratio of monosaccharides to oligosaccharides was 1:9 and the main component, xylose, was 85.9% in content. The total recovery rate of hemicellulose was 80%. Water washed materials were subsequently extracted with ethanol. The optimum extraction conditions in this work were 40% ethanol, fiber/liquor ratio 1:50 (w/v), severity log(R)=3.657 (180 degrees C for 20 min), 0.1% NaOH. The lignin yield was 75% by acid precipitation and 85% ethanol solvent was recovered. The lignin was purified using Bj?rkman method. Infrared spectrometry (IR) results indicated that the lignin belonged to GSH (guaiacyl (G) syringyl (S) and p-hydroxyphenyl (H)) lignin and its purity rate reached 85.3%. The cellulose recovery rate was 94% and the results of electron spectroscopy for chemical analysis (ESCA) and infrared spectrometry (IR) showed that hemicellulose and lignin content decreased after steam explosion and ethanol extraction. 相似文献
7.
Lignin, the second most abundant biopolymer on earth and with a predominantly aromatic structure, has the potential to be a raw material for valuable chemicals and other bio-based chemicals. In industry, lignin is underutilized by being used mostly as a fuel for producing thermal energy. Valorization of lignin requires knowledge of the structure and different linkages in the isolated lignin, making the study of structure of lignin important. In this article, lignin samples isolated from two types of reactors (autoclave reactor and displacement reactor) were analyzed by FT-IR, size exclusion chromatography, thermogravimetric analysis (TGA), and Py-GC-MS. The average molecular mass of the organosolv lignins isolated from the autoclave reactor decreased at higher severities, and FT-IR showed an increase in free phenolic content with increasing severity. Except for molecular mass and molecular mass dispersity, there were only minor differences between lignins isolated from the autoclave reactor and lignins isolated from the displacement reactor. Carbohydrate analysis, Py-GC–MS and TGA showed that the lignin isolated using either of the reactor systems is of high purity, suggesting that organosolv lignin is a good candidate for valorization. 相似文献
8.
Julien Bauwens Cédric Tarayre Catherine Brasseur Christel Mattéotti Jacqueline Destain Micheline Vandenbol Daniel Portetelle Philippe Thonart Edwin De Pauw Eric Haubruge Frédéric Francis 《Symbiosis (Philadelphia, Pa.)》2012,58(1-3):119-125
The gut of lower termites is populated by numerous microbial species belonging to prokaryotes, fungi, yeasts and protists. These micro-organisms are organized in a complex symbiotic system, interacting together and with the insect host. Their likely ability to degrade ligno-cellulosic compounds could lead to improvements in second generation biofuels production. Lignin elimination represents a critical point as this polymer significantly interferes with industrial process of cellulose. Although host produces its own lignin-degrading enzymes, some symbionts may participate in digestion of lignin and its degradation products in termite gut. Here, we compared gut proteomes from R. santonensis after rearing on artificial diets composed of cellulose with and without lignin. The effect of lignin in artificial diets on different parts of the digestive tract was compared through liquid chromatography associated with tandem mass spectrometry (LC-MS/MS) experiments. Enzymatic assays were performed to characterize activities present in R. santonensis digestive tract after feeding on artificial diets. Microscopic observations of microbial communities provided some information on population balances after feeding experiment. 相似文献
9.
Separation of brain monosialoganglioside molecular species by high-performance liquid chromatography 总被引:1,自引:0,他引:1
A method for the separation of molecular species of brain monosialogangliosides by high-performance liquid chromatography is described. GM4, GM3, GM2, and GM1 were purified from human brain and their individual molecular species were separated on a C18 reversed-phase column. Peaks were identified by mass spectrometry of the intact ganglioside, by gas-liquid chromatography of the fatty acids, and by high-performance liquid chromatography of the long chain bases. A characteristic elution sequence of molecular species permitted their identification based upon their retention times on the reversed-phase column. 相似文献
10.
N4-Hexanoylspermidine,a New Polyamine-Related Compound That Accumulates during Ovary and Petal Senescence in Pea 总被引:1,自引:1,他引:0
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Perez-Amador MA Carbonell J Navarro JL Moritz T Beale MH Lewis MJ Hedden P 《Plant physiology》1996,110(4):1177-1186
A previously unknown polyamine conjugate that accumulates in senescing ovaries of pea (Pisum sativum L.) was shown by mass spectrometry, nuclear magnetic resonance, and chemical synthesis to be N4-hexanoylspermidine (hexanoyl-spd) This structure was indicated by analysis of the dansylated polyamine using fast atom bombardment mass spectrometry, following purification by high-performance liquid chromatography. Furthermore, acid hydrolysis of the compound yielded spermidine and hexanoic acid. 1H-nuclear magnetic resonance suggested that spermidine was substituted at N4 in the conjugate. Hexanoyl-spd was synthesized, and its didansyl derivative was shown to have an identical mass spectrum and high-performance liquid chromatography retention time as the derivatized natural compound. Further confirmation of its structure was obtained by comparison of the synthetic and natural polyamines as trifluoroacetyl derivatives using gas chromatography-mass spectrometry. This new polyamine conjugate is present in pea ovaries at low levels at anthesis and its concentration remains low in developing seeded fruit or in parthenocarpic fruit that have been induced by application of growth regulators to emasculated flowers or by topping the plant. Conjugate levels are also low in parthenocarpic fruit induced naturally in the slender (la crys) mutant. However, levels of hexanoyl-spd increase progressively in senescing petals and ovaries, beginning at anthesis or 2 d later, respectively. 相似文献
11.
Vater J Kablitz B Wilde C Franke P Mehta N Cameotra SS 《Applied and environmental microbiology》2002,68(12):6210-6219
An innovative method was developed for rapid sensitive detection and efficient structural characterization of lipopeptide biosurfactants by matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry by using whole microbial cells and crude culture filtrates as targets in combination with surface tension measurements. This was done for a bacterial strain that was isolated from petroleum sludge and efficiently produces biosurfactants. This organism was identified by using biochemical, physiological, and genetic parameters as a Bacillus subtilis strain, designated B. subtilis C-1. This assignment was supported by a mass spectrometric investigation of the secondary metabolite spectrum determined by whole-cell MALDI-TOF mass spectrometry, which revealed three lipopeptide complexes, the surfactins, the iturins, and the fengycins, which are well-known biosurfactants produced by B. subtilis strains. These compounds were structurally characterized by in situ structure analysis by using postsource decay MALDI-TOF mass spectrometry. The isoforms were separated by miniaturized high-resolution reversed-phase high-performance liquid chromatography for mass spectrometric characterization. Iturin compounds which contain unusual fatty acid components were detected. 相似文献
12.
H Kitagawa H Nakada S Fukui I Funakoshi T Kawasaki I Yamashina S Tate F Inagaki 《Biochemistry》1991,30(11):2869-2876
We have isolated four novel oligosaccharides with the sialyl-Lea structure from human milk using a monoclonal antibody, MSW 113. These oligosaccharides were purified by affinity chromatography on a column of the immobilized monoclonal antibody and by high-performance liquid chromatography. The results of structural analyses, i.e., 500-MHz 1H NMR spectroscopy, fast atom bombardment mass spectrometry, and binding to specific anticarbohydrate antibodies, are consistent with the following structures. (formula; see text) 相似文献
13.
Manfred R. Moeller 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》1992,580(1-2):125-134
This article reviews the analysis of 31 drugs and drug metabolites in human hair by thin-layer chromatography, high-performance liquid chromatography, gas chromatography, gas chromatography—mass spectrometry and mass spectrometry. The most important detection method after chromatographic separation of the components is the mass spectrometry because of its sensitivity and specifity. Washing steps to exclude external contamination, extraction, derivatization, stationary phases, detection modes and detection limits of the mass spectrometric and gas chromatographic-mass spectrometric procedures are presented in five tables. Additionally, a method for a gas chromatographic-mass spectrometric screening procedure is presented. 相似文献
14.
O N Jensen P Roepstorff B Rozynov M Horanyi J Szelenyi S R Hollan E A Aseeva V A Spivak 《Biological mass spectrometry》1991,20(10):579-584
S-Aminoethylated-alpha A and -beta A globin tryptic peptides separated by reversed-phase high-performance liquid chromatography have been analysed by plasma desorption mass spectrometry. Almost all the expected alpha A and beta A tryptic fragments were tentatively assigned relative to the known globin chain sequences based on the molecular weight obtained by plasma desorption mass spectrometric analysis of the purified peptides. The application of plasma desorption mass spectrometry for structure elucidation of a haemoglobin alpha-chain variant revealed the first case of Hb Hasharon in Hungary. 相似文献
15.
Meng-lei Huan Si-yuan Zhou Zeng-hui Teng Bang-le Zhang Xin-you Liu Jie-pin Wang Qi-bing Mei 《Bioorganic & medicinal chemistry letters》2009,19(9):2579-2584
The synthetic DOX–LNA conjugate was characterized by proton nuclear magnetic resonance and mass spectrometry. In addition, the purity of the conjugate was analyzed by reverse-phase high-performance liquid chromatography. The cellular uptake, intracellular distribution, and cytotoxicity of DOX–LNA were assessed by flow cytometry, fluorescence microscopy, liquid chromatography/electrospray ionization tandem mass spectrometry, and the tetrazolium dye assay using the in vitro cell models. The DOX–LNA conjugate showed substantially higher tumor-specific cytotoxicity compared with DOX. 相似文献
16.
Michal Svoboda Michael Przybylski Jolanda Schreurs Atsushi Miyajima Kenneth Hogeland Max Deinzer 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》1991,562(1-2)
The primary structure of Baculovirus-expressed mouse interleukin-3 produced in infected Bombyx mori larvae was characterized by liquid secondary ion mass spectrometry and 252Cf-plasma desorption mass spectrometry in combination with selected protein microchemical reactions. Interleukin-3 was found to consist of at least two glycoprotein species of ca. 17 000 dalton. Characterization of tryptic and S. aureus V8 protease peptides by Edman degradation combined with plasma desorption mass spectrometry showed that two N-glycosylation sites, Asn-16 and Asn-86, were present. N-Glycan residues were shown by liquid secondary ion mass spectrometry and high-performance liquid chromatography to consist of mannose, fucose, and glucosamine. The presence of galactosamine indicated that O-glycosylated residues were present, in addition to the N-glycosylated residues. Glucose was also present, which indicated incomplete processing of the insect-expressed N-linked oligosaccharides. 相似文献
17.
O S Reshetova B V Rozynov M V Bezrukov I A Bogdanova 《Bioorganicheskaia khimiia》1986,12(12):1625-1636
Mass spectrometry has been applied to the structural analysis of one of the glycopeptides from blastolysin, antitumor bacterial preparation isolated from the Lactobacillus bulgaricus cell wall. The glycopeptide (MW 10,000) was subjected to partial acid hydrolysis (6 N HCl, 100 degrees C) and the resulting products were dansylated or trifluoroacetylated and methylated or deuteromethylated. The mixture of these derivatives was examined by high-performance liquid chromatography or gas chromatography followed by mass spectrometry using electron impact and ammonia chemical ionization techniques. 相似文献
18.
Albrecht Grupe Gerhard Spiteller 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》1981,226(2)
A sequence of chromatographic methods (thin-layer chromatography, high-performance liquid chromatography and glass capillary gas chromatography) was used to separate the acid fraction of human urine. The power of this method to separate and detect previously unknown compounds and the elucidation of their final structure with mass spectrometry is exemplified by the identification of N-acetyl-2-aminooctanoic acid as a metabolic compound in the urine of healthy individuals.In addition, the conjugate of glycine with indolepropionic acid, N-formylanthranilic acid, succinoylphenylalanine, δ-hydroxyvaleric acid, δ-hydroxycapric acid, 3-hydroxyadipic acid, and higher homologues were detected in a polar fraction of human urine. 相似文献
19.
Fountain KJ Gilar M Budman Y Gebler JC 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》2003,783(1):61-72
Singly- and dually-labeled synthetic oligonucleotides were purified by ion-pair reversed-phase high-performance liquid chromatography using a 50x4.6-mm column packed with porous, 2.5 micrometer C(18) sorbent. We studied the mechanism of dye-labeled oligonucleotide retention in order to improve the quality of purification. By-products of oligonucleotide synthesis were characterized by liquid chromatography with mass spectrometry detection (LC-MS). We purified oligonucleotides labeled with 6-carboxyfluorescein (6FAM), hexachlorofluorescein (HEX), tetrachlorofluorescein (TET), carboxytetramethylrhodamine (TAMRA) and indodicarboxycyanine (Cy3) dyes, as well as dually-labeled TaqMan probes. Purification of a 0.1-micromole oligonucleotide synthesis in a single injection was demonstrated. 相似文献
20.
木质素降解产物对微生物产生的抑制作用,是燃料乙醇生物炼制的主要瓶颈之一。本文以树干毕赤酵母为发酵菌株,研究木质素降解产物中3种酚酮类(4-羟基苯乙酮、4-羟基-3-甲氧基苯乙酮、4-羟基-3,5-二甲氧基苯乙酮)对其木糖乙醇发酵及酵母细胞脂肪酸组成的影响。采用高效液相色谱(HPLC)和气相色谱-质谱联用(GC/MS)技术对乙醇发酵性能和酵母细胞脂肪酸组成进行分析。研究结果表明,酚酮类物质对乙醇发酵呈现抑制作用且其分子量越低抑制作用越明显,当4-羟基苯乙酮浓度为1.50 g/L时,发酵24 h的木糖利用率、乙醇得率和乙醇浓度分别下降了42.47%、5.30%和9.76 g/L;培养基中存在酚酮类物质时,酵母细胞中的不饱和脂肪酸的比例上升,添加1.50 g/L的3种酚酮类物质后,树干毕赤酵母细胞不饱和脂肪酸和饱和脂肪酸的比例从原来的2.58分别上升到3.03、3.06和3.61,酵母细胞膜的流动性随之上升,不稳定性提高。因此,酚酮类物质能够降低酵母生长、提高不饱和脂肪酸的比例以及降低乙醇发酵能力,有效降低或去除木质素降解产物含量是提高木质纤维原料生物炼制的关键。 相似文献