Determination of aminophenol isomers by high-speed liquid chromatography

The simultaneous quantitation of aminophenol isomers (p-, m- and o-aminophenol) and anilline, which with previously reported methods was difficult and/or complicated, has been performed using a high-speed liquid chromatographic method. The resolutions of chromatograms were appropriate for separation and quantitation, when analysis times were 20 minutes. Since the method is applicable in aqueous media, a successful quantitation of p- and o-aminophenol formed by cytochrome P-450 model systems was carried out.     

Recognition of chitin and proteins in invertebrate cuticles using analytical pyrolysis/gas chromatography and pyrolysis/gas chromatography/mass spectrometry

Flash pyrolysis/gas chromatography (py/GC) and py/GC/mass spectrometry (MS) have been utilized to characterize the cuticles of invertebrates chemically. Pyrolysis products have been identified and assigned to specific cuticular components. Acetylpyridones, acetamidofuran, 3-acetamido-5-methylfuran and 3-acetamido-(2 and 4)-pyrones are proposed as characteristic pyrolysis markers for chitin. Pyrolysis products displaying ions of m/z 70, 154, 168, 194 are thought to derive from diketopiperazine structures and provide potential markers for proteins and peptides in which proline, alanine, valine, arginine and glycine are the dominant amino acids. These products, constituting specific pyrolysis markers for invertebrate cuticles, may reflect the amino acid composition of their constituent structural proteins. The source of the various pyrolysis products of proteins has been verified by pyrolysis of reference proteins, peptides and amino acid mixtures. The presence of additional pyrolysis products related directly to histidine and catechol moieties is consistent with the chemical structure and composition proposed for arthropod cuticles based on recent work utilizing solid state 13C and 15N nuclear magnetic resonance. This study constitutes the first comprehensive chemical characterization of the pyrolysis products of invertebrate cuticles and provides the basis for future investigations requiring qualitative screening for cross-linked chitin and proteins in modern and fossil cuticles and in materials, e.g. geopolymers, that may be derived from them.     

Determination of an anti-inflammatory methanesulfonanilide in plasma by high-speed liquid chromatography

A sensitive and chemically specific high-speed liquid chromatographic method was developed for the determination of 4-nitro 2-phenoxymethanesulfonanilide in plasma. The method includes selective extraction of the anti-inflammatory drug and an internal standard, 2-(4'-chlorophenoxy)-4-nitromethanesulfonanilide, into benzene from acidified plasma followed by reextraction into 0.2 N NaOH. The aqueous layer is acidified, and the drug is reextracted into benzene. The benzene is evaporated, and the residue is dissolved in a small volume of acetonitrile. A 10-microliter aliquot is analyzed on a reversed-phase column. The mean overall extraction recovery, after correction for aliquot factors, is 99%. The accuracy, expressed as the relative error, is 4, 0.3, and -3% at 0.60, 1.50, and 3.00 microgram/ml, respectively. Repeated analysis of reference standards indicates that the precision, expressed as the relative standard deviation, is 3% or less. The lower sensitivity limit is 0.2 microgram/ml with a 2-ml plasma sample. The method was applied successfully to the determination of plasma levels of 4-nitro-2-phenoxymethanesulfonanilide in humans and rats in metabolic experiments at pharmacological doses.     

High-performance liquid chromatography of proteins

Incubation of liver microsomes with GDP [14C] mannose leads to the formation of lipid-linked derivatives of [14C] mannose, a dolichol phosphate monosaccharide and dolichol pyrophosphate oligosaccharides. Standard procedures for separating these two types of compounds from each other were found to be deficient in that fractions thought to contain only dolichol pyrophosphate oligosaccharides are contaminated with dolichol phosphate mannose. This paper presents a column chromatographic procedure which conveniently separates the products of an 8 min labeling experiment into two components; dolichol phosphate [14C]mannose and a [14C]-mannose containing oligosaccharide which is also lipid bound. When this oligosaccharide is released from the lipid by hydrolysis and chromatographed on Sephadex G-50 or G-15 it gives a single peak with an indicated molecular weight of 1100. However, when this released oligosaccharide is chromatographed on concanavalin A Sepharose it is resolved into two peaks suggesting that there may be 2 oligosaccharide of approximately the same size but different structures. After brief periods of labeling with GDP [14C]mannose (5 s) an additional oligosaccharide of 3 to 4 sugar residues can be found in the dolichol pyrophosphate oligosaccharides fraction. Incubation of liver microsomes with UDP [14C]glucose or UDP[14C]galactose produces oligosaccharide components containing 7--8 sugar residues. Labeling of microsomes with UDP[14C]acetylglucosamine gives rise to three different components, including a lipid bound oligosaccharide containing 3- 5 sugar residues.     

Analytical development of electrospray and nanoelectrospray mass spectrometry in combination with liquid chromatography for the characterization of proteins

Microbial pathogens may be transmitted to humans via food animals and food animal products. A quick reference table is presented to provide easy access to food safety information related to the major food animal product areas. Included in the table are the pathogens, mode of transmission, public health impact, and control and prevention strategies for poultry, beef, dairy products, and pork.     

Emulsification and mass transfer in ladle metallurgy

Measurements of slag emulsification in gas-stirred ladles were carried out in cold-model systems of different geometric sizes. Detaching of slag droplets – necessary for efficient emulsification – only takes place if the flow velocity at the slag/metal interface exceeds a certain level. The use of a centric nozzle leads at high gas flow rates to considerably larger degrees of emulsification than eccentric stirring. The reason of this phenomenon is that the recirculation flow during centric gas injection transports larger amounts of emulsified droplets into deeper regions of the melt while during eccentric stirring there is more time for reseparation of slag droplets into the top slag. Comparing emulsification results with mass-transfer measurements, the dependence between rate constants as well as degrees of emulsification and Froude number shows similar behaviour.     

Oxygen mass transfer calculations in large arteries

Parkinsonism is characterised by overactive glutamatergic transmission in the cortico-striatal and subthalamo-medial pallidal pathways. Local blockade of glutamatergic transmission in these pathways can alleviate parkinsonian symptoms. The effectiveness of the treatment, however, is often limited by the simultaneous appearance of unwanted side-effects. These side-effects, including ataxia and dissociative anaesthesia, are particularly problematic when N-methyl-D-aspartate (NMDA) antagonists are used. In an attempt to overcome these problems we have attempted to manipulate excitatory amino acid (EAA)-mediated neurotransmission indirectly by targeting the NMDA receptor associated modulatory sites. We review evidence which demonstrates that antagonists for both the NMDA associated glycine and polyamine sites can reverse parkinsonian symptoms when injected intra-cerebrally in both MPTP-treated and bilateral 6-OHDA lesioned marmosets without eliciting unwanted side-effects. We further review preliminary data which suggest that ifenprodil, a polyamine site antagonist, has striking anti-parkinsonian actions in the marmoset. Potential mechanisms of action underlying these effects are discussed in terms of NMDA receptor subtypes and the neuroanatomical locus of action. The anti-parkinsonian efficacy of intra-striatally administered EAA antagonists leads us to question the view of dopamine acting in the striatum as a simple neuromodulator.     

Triplet state energy transfer in several proteins

Energy transfer between excited triplet states of aromatic amino acid residues was observed at 1.4 degrees K. The distance necessary for energy transfer between monomeric tyrosine and tryptophan residues was determined to be roughly 63 A. Total phosphorescence decay rate constants for several proteins were determined while emission corresponding to tyrosine and tryptophan residues was monitored. The observed decay rate constants are interpreted in terms of intramolecular interactions of the polypeptide residues.     

The determination of methionine in proteins by gas-liquid chromatography

Intact methionine residues in proteins were rapidly and precisely determined by measuring methyl thiocyanate released during the reaction with CNBr and separated by g.l.c. Conditions for the reaction and for chromatography on columns of Porapak P-S are described. The recovery of methyl thiocyanate from several methionine derivatives and analogues were examined. Carbamoylmethionine was adopted as a stable primary standard and ethyl thiocyanate as internal standard. The measured methionine content of several isolated proteins was close to the theoretical value indicated by previous work and the results for these and a range of food proteins agreed well with results obtained by ion-exchange chromatography after performic acid oxidation. Since CNBr does not react with methionine sulphoxide and a preliminary hydrolysis is not required, the method discriminates between methionine and any methionine sulphoxide that may be present. It could be useful in studies on the nutritional availability of methionine in processed foods.     

Distinguishing small molecular mass differences of proteins by mass spectrometry

This study was undertaken to determine if acidic or basic fibroblast growth factor (FGF1 or FGF2) or vascular endothelial growth factor (VEGF) alters the radiation response of small bowel after total-body irradiation (TBI). Female C3H mice were treated with various doses of angiogenic growth factor administered intravenously 24 h before or 1 h after TBI. Radiation doses ranged from 7 to 18 Gy. End points measured were the number of crypts in three portions of the small bowel, the frequency of apoptosis of crypt cells at various times after TBI, and the LD50/30 (bone marrow syndrome) and LD50/6 (GI syndrome). Fibroblast growth factors alone, without TBI, decreased the number of crypts per circumference significantly. Among the factors tested, FGF2 caused the greatest decline in baseline crypt number. Despite this decrease in the baseline crypt number, after irradiation the number of surviving crypts was greater in animals treated with growth factor. The greatest radioprotection occurred at intermediate doses of growth factor (6 to 18 pg/mouse). Mice treated with FGF1 and FGF2 had crypt survival curves with a slope that was more shallow than that for saline-treated animals, indicating radiation resistance of crypt stem cells in FGF-treated mice. The LD50/6 was increased by approximately 10% for all treatments with angiogenic growth factors, whether given before or after TBI. Apoptosis of crypt cells was maximum at 4 to 8 h after TBI. The cumulative apoptosis was decreased significantly in animals treated with angiogenic growth factors, and the greatest protection against apoptosis was seen in animals treated with FGF2 prior to TBI. All three angiogenic growth factors tested were radioprotective in small bowel whether given 24 h before or 1 h after irradiation. The mechanism of protection is unlikely to involve proliferation of crypt stem cells, but probably does involve prevention of radiation-induced apoptosis or enhanced repair of DNA damage of crypt cells.     

Determination of serum retinol (vitamin A) by high-speed liquid chromatography

We report a fast and simple high-speed liquid-chromatographic assay for serum retinol. Only 100 mul of serum is required. The lower detection limit is 50 mug/liter; linearity was demonstrated up to 1.50 mg/liter. On analyzing a serum pool eight times, a CV of 2.5% was obtained. Values by this method are compared with results obtained by a flurometric method [Clin. Chem. 16, 766(1970)].     

High-performance liquid chromatography of amino acids, peptides and proteins. CXXXIX. Impact of operating parameters in large-scale chromatography of proteins

Large-scale chromatography has been playing an important role in downstream treatment processing in biotechnology. In order to improve the productivity, the throughput of the chromatographic equipment was often increased by increasing the flow-rate and/or by increasing the column sample loading. This paper reports the results of a study on the impact of these and other operating parameters in affinity and ion-exchange chromatographic columns when used for protein purification. A sectional model was developed to predict protein adsorption processes in a packed column. The formulations of this mathematical model are presented in the Appendix. The present study was carried out with computer simulation based on this model and using data obtained from laboratory-scale columns. This model can simulate both the adsorption and washing stages of the protein purification process for both porous and non-porous particles. The effects of changing operating parameters were simulated and contour plots were generated for the easy identification of these effects. It was shown that both flow-rate and column loading can have a considerable impact on the processing rate and the yield of the column. As for the column capacity utilization, the impact of changing flow-rate is not significant at column loading of less than 80% in the test case. It was suggest that the present investigation provides a systematic predictive strategy which will greatly reduce the need for expensive, labour-intensive and time-consuming experimental work during process scale-up.     

Electrical conduction and mass transfer in crystalline oxides

The electrical conductivity of oxides is closely related to the reducibility or oxidizability of their metals. The experimental results of the authors and reported data are used to explain the behavior of the electrical conductivity of solid oxides and its relation to mass transfer with allowance for the ionic structure of these oxides.     

Model experiments on mass transfer in ladle metallurgy

Model experiments on mass transfer in gas-stirred ladles were carried out in reactors of different geometric dimensions. The model system consists of: water, cyclohexan as model slag, iodium as element to be extracted from water into slag phase, and compressed air as stirring gas. The experimental results show that when using an eccentric bottom nozzle, rate constants of mass transfer are always smaller than with centric gas injection. Centric stirring leads to comparatively larger increases of rate constants if a certain gas flow rate is exceeded. Both results can be explained by different emulsification conditions of slag phase. Theoretical calculations of residence times show that mainly the emulsification of small droplets taken along by the recirculation flow is responsible for accelerations of mass transfer in gas-stirred ladles.     

Interfacial phenomena and mass transfer in extractive metallurgy

Interfacial phenomena play an important role in pyrometallurgical processes and knowledge of them and of physical properties involved is necessary for understanding the mechanisms and the kinetics of such reactions. A large number of measurements, performed at Irsid under equilibrium conditions, are presented in this review. Several experimental techniques were used and more particularly: – Sessile drop method for measurement of liquid metals surface tension and contact angle liquid metal/solid oxide; – measurement of the contact angle between a liquid slag drop and its liquid metal substrate from which the interfacial tension can be derived; – direct determination of the interfacial tension from X-ray pictures of metal drops immersed in the slag. The systems studied consisted, for the metal phase, of binary and ternary Fe alloys containing C, Mn, Si, O, S and, for the slag phase, binary and ternary mixtures made from CaO, SiO₂, Al₂O₃, MnO, iron oxides, CaF₂ and Na₂O. A strong effect of O and S potentials was observed. For non-equilibrium conditions, however, the dynamic interfacial tension between liquid metal and slag decreases sharply when an intense mass transfer occurs through the interface. The potential consideration of interfacial turbulence phenomena (Marangoni effect) in metallurgical reactions is also discussed.     

冷却工艺对高速连铸结晶器传热的影响

为了分析冷却水的供水工艺对结晶器铜壁和冷却水温度场的影响,基于结晶器铜壁热电偶实测温度,构建铸坯/铜壁传热反问题和铜壁/冷却水正问题数学模型,采用ANSYS建立铸坯/铜壁/冷却水数值分析模型,对薄板坯结晶器温度场进行耦合传热分析,解析不同冷却工艺对高速薄板坯连铸结晶器内传热行为的影响.结果表明,水缝内冷却水流动方向对铜...