Separation and characterisation of products of the reaction between propionamide and formaldehyde

Summary Propionamide-formaldehyde reaction products have been characterised using micellar electrokinetic chromatography (MEKC), HPLC-electrospray-MS (HPLC-ES-MS) and time-of-flight-MS (TOF-MS). HPLC-MS-MS was used to distinguish between isomeric species. The MEKC separation of all reaction products was obtained with good resolution and efficiency. Comparison of water-micelle distribution constants (P _MEKC) for propionamide-formaldehyde with those for caprolactam-formaldehyde reaction products suggests that the concentration of sodium dodecyl sulfate appropriate for use in MEKC is inversely related toP _MEKC. Quantification of all species containing one or two amide units was achieved using standard calibration, mass balance and the assumption of identical absorption coefficients for the same functional groups in monomers and dimers.     

Separation and determination of m-phenoxybenzaldehyde reaction products by capillary gas chromatography

A capillary GC method employing an internal standard has been developed and successfully used for quantitative determination both of the raw materials used for the manufacture of m-phenoxybenzaldehyde and for the components of the reaction mixtures obtained at various stages of the development of the process. A complete analysis can be performed in a single temperature programmed run.     

Separation and partial characterization of Maillard reaction products by capillary zone electrophoresis

Capillary zone electrophoresis proved useful for separating small amounts of both charged and uncharged solutes that are otherwise difficult to analyse. A typical complex mixture that had previously resisted all analytical approaches, including reversed-phase separations, is the products arising from the reaction of free amino acids with aldehydic sugars (Maillard reaction products). By using capillary zone electrophoresis [untreated capillary 50 cm x 75 microns I.D., 18 kV, 0.02 mol/l phosphate buffer (pH 7.5)], a number of products resulting from the reaction of glucose or ribose with glycine, alanine and isoleucine were separated and partially characterized. They were separated (1) without derivatization (and profiles of compounds absorbing at 220 nm were obtained), (2) as phenylthiocarbamyl derivatives in a search for reactive amino groups and (3) after derivatzation with 2,4-dinitrophenylhydrazine in a search for a method for compounds with a free aldehydic group. Phenylthiocarbamyl derivatives were separated in 0.005 mol/l borate buffer (pH 9.6) at 20 kV and 25 microA. Separation of 2,4-dinitrophenylhydrazones was effected by electrokinetic micellar chromatography in the same apparatus using a 50 cm x 75 microns I.D. capillary at 10 kV in 0.01 mol/l Na2HPO4-0.006 mol/l tetraborate, 0.050 mol/l with respect to sodium dodecyl sulphate. The results are compared with those given by high-performance liquid and thin-layer chromatography.     

Separation and analysis of DNA sequence reaction products by capillary gel electrophoresis

This paper demonstrates the potential of capillary gel electrophoresis with laser induced fluorescence detection as a tool for DNA sequence determination. Both synthetic oligonucleotides and single-stranded phage DNA were utilized as templates in the standard chain termination procedure. Primer molecules were tagged at the 5' end with the fluorescent dye, JOE. First, baseline resolution of a dA extended primer from 18 to 81 bases long, a total of 64 fragments, was observed. A second synthetic template was designed to yield alternating stretches of dA and dT extensions of the primer. Thirdly, the sequence reaction products from a synthetic oligonucleotide template containing all four bases was analyzed in four independent runs, one for each of the four base-specific reactions. In all cases, the expected number and patterns of peaks were observed by capillary gel electrophoretic analysis. Finally, separation of sequence reaction products generated with single-strand M13mp18 phage DNA as template exhibited baseline resolution of fragments differing in length by a single nucleotide and from 18 to greater than 330 bases total length.     

环十二碳酮与苯基乙二醇反应产物的分离及确定

环十二碳酮(1)与苯基乙二醇(2)在对甲苯磺酸催化下的反应粗产物具有浓郁的花香,最初我们以为这种香气来源于缩酮2-苯基-1,4-二氧杂螺[4,11]十六烷(3):但该反应粗产物经反复重结晶后得到的无色晶体却没有香气、经红外、~1H核磁等谱图及碳氢分析鉴定该无色晶体确是缩酮3.既然缩酮3没有气味,则反应粗产物的香气必然来自反应中的副产物。薄层色谱分析表明,该反应中副产物较多。我们猜测,副反应可能主要来自2。     

Separation of the reaction products of cyanazine and terbuthylazine nitrosation by thin-layer chromatography

N-Nitroso-triazine herbicide species were synthesized as reference standards and their reaction products were separated by TLC. The method was established for analytical as well as for preparative amounts to achieve a sufficient separation between the educts, the byproducts and the N-nitrosated moieties. Separation of the reaction mixture was performed by two-dimensional TLC, using different mobile phases for each dimension. In the second dimension two developments were executed using the same solvent mixture. To examine the quality of performance, the separation zones were scrapped off, extracted and analyzed in the HPLC-diode array detection (DAD) mode in comparison with standards of the educts and the products of their N-nitrosation. This method minimizes the contact with the resulting N-nitroso compounds. At present nothing is known about the toxicological relevance or risk assessment on human health of N-nitrosocyanazine or N-nitrosoterbuthylazine species. Therefore the practical handling of these putative mutagenic and carcinogenic substances should be seen under the aspect of precaution and prevention of contamination. Received: 25 August 1997 / Revised: 13 October 1997 / Accepted: 21 October 1997     

Separation of the reaction products of cyanazine and terbuthylazine nitrosation by thin-layer chromatography

N-Nitroso-triazine herbicide species were synthesized as reference standards and their reaction products were separated by TLC. The method was established for analytical as well as for preparative amounts to achieve a sufficient separation between the educts, the byproducts and the N-nitrosated moieties. Separation of the reaction mixture was performed by two-dimensional TLC, using different mobile phases for each dimension. In the second dimension two developments were executed using the same solvent mixture. To examine the quality of performance, the separation zones were scrapped off, extracted and analyzed in the HPLC-diode array detection (DAD) mode in comparison with standards of the educts and the products of their N-nitrosation. This method minimizes the contact with the resulting N-nitroso compounds. At present nothing is known about the toxicological relevance or risk assessment on human health of N-nitrosocyanazine or N-nitrosoterbuthylazine species. Therefore the practical handling of these putative mutagenic and carcinogenic substances should be seen under the aspect of precaution and prevention of contamination.     

Permethylation of barbiturates. Separation and characterization of the reaction products by gas chromatography-mass spectrometry

A Series of barbiturates (I) has been permethylated with the methylsulfinylmethide carbanion and methyl iodide and the products (II to IV) separated, quantified and characterized by g.c.-m.s. The ratios and fragmentation patterns of II to IV are highly dependent upon the nature of the R-groups. All fragment ions have been confirmed by accurate mass measurements or perdeuteriomethylation.     

Structural and EPR characterisation of single electron and alkyl transfer products from reaction of dimethyl magnesium with bulky alpha-diimine ligands

Treatment of dimethylmagnesium with bulky alpha-diimine ligands provides either the biradical methyl-bridged complexes [(alpha-diimine-.)Mg+(mu-CH3)]2 via single electron transfer (SET), or the product of methyl transfer to an imine carbon atom depending upon conditions.     

Separation of protactinium from uranium and other reaction products in the reaction of 60 MeV/nucleon 18O with natural uranium

Protactinium was produced by the reaction of 60 MeV/nucleon ¹⁸O with natural uranium. A simple, relatively fast radiochemical procedure was developed, which can be used for the extraction separation of protactinium from uranium and from the complex reaction products using 1-phenyl-3-methyl-4-benzoyl-5-pyrazolone and tri-iso-octylamine as extractants. Measurements of the gamma-ray spectra for the separated protactinium fractions were performed with a HPGe detector. The measured g-ray spectrum of protactinium shows that the decontamination from the main impurity elements, especially zirconium and niobium, is quite satisfactory.     

Separation and confirmation of the product and by products of the reaction of cyclododecanone with phenyl ethylene glycol

It has been proved, by TLC and synthetic methods, that the odoriferous material formed in the reaction of cyclododecanone and phenyl ethylene glycol under acidic condition, was not the ketal of cyclododecanone, but an acetal of phenylacetaldehyde with phenyl ethylene glycol—the by-product formed from phenyl ethylene glycol. The cis-trans isomers of the acetal were separated by chromatography, the configurations were confirmed via the analysis of their ¹H NMR spectra, and the odors were evaluated as well.     

Separation and characterisation of sphingoceramides by high-performance liquid chromatography-electrospray ionisation mass spectrometry

We developed a simple and reliable analytical method for the quantification and the characterization of ceramides extracted from biological samples by high-performance liquid chromatography (HPLC) coupled to electrospray ionisation tandem mass spectrometry (ESI/MS/MS). The chromatographic separation of analytes was carried out in a RP8 column, eluting with a methanol-water mixture in gradient elution mode. The separated lipids were detected by total ion monitoring and characterised by MS/MS spectra; quantitative analysis was performed by integrating the extracted ion peaks obtained in the negative ion mode. Good repeatability was obtained for retention time (0.3-2%), peak area ratio (A(S)/A(IS), 2-8%), as well as limit of detection (LOD, 5-26 pg) and quantification (LOQ, 13-53 pg). The method was validated for the analysis of N-palmitoyl-D-erythro-sphingosine (Cer16), N-stearoyl-D-erythro-sphingosine (Cer18), N-tetracosanoyl-D-erythro-sphingosine (N24:0, lignoceric ceramide, Cer24:0), and N-tetracos-15'-enoyl-D-erythro-sphingosine (N24:1, nervonic ceramide, Cer24:1), giving good results. Lipid mixtures, extracted from skin and epidermal cells, were analysed for their content of the studied ceramides.     

Cross-species characterisation of abundantly expressed Ochrobactrum anthropi gene products.

The identity of 45 protein spots representing 32 orthologues within the Ochrobactrum anthropi proteome within a gradient of pH 4-7, and mass range 5-90 kDa were determined across species boundaries. These proteins could be classified into 13 functional categories and establish metabolic, regulatory and translatory systems including amino acid biosynthesis, electron transport and the potential for plant symbiosis in a molecularly understudied organism. Amino acid composition and/or peptide mass fingerprinting were employed as a means to search the Swiss-Prot and OWL protein sequence databases for similarity within a broad taxonomic class of bacteria. Candidate matches from database searches could be compared and a simple multiplication matrix based on co-occurrence and rank within the top 96 most similar entries was used to provide statistical confidence. This mathematical matrix was evaluated with respect to the characterisation of O. anthropi, an unsequenced and understudied bacterium, in the light of the recent influx of DNA sequence information.     

Click synthesis by Diels-Alder reaction and characterisation of hydroxypropyl methylcellulose-based hydrogels

The Diels-Alder reaction was used to fabricate hydroxypropyl methylcellulose-based hydrogels. First, hydroxypropylmethylcellulose (HPMC) was modified by a carboxyl-containing diene molecule (SFA) which was synthesised from furfurylamine and succinic anhydride. Second, dienophile groups were introduced into HPMC by the coupling reaction with N-maleoyl alanine (AMI) using N,N′-dicyclohexylcarbodiimide (DCC) and 4-dimethylaminopyridine (DMAP). Subsequently, the asprepared furan- and maleimide-modified HPMC were dissolved in water and gelation was observed at a pre-determined temperature after a period of time. The samples thus obtained were characterised by FTIR, NMR, SEM, etc. The gelation time changing with temperature, concentration of the solution, and solvent was measured. It was found that gelation time decreased with increasing temperature and concentration of the solution, and that water had a rate-accelerating effect on Diels-Alder reaction. The swelling behaviour indicates that the hydrogels have a high swelling ratio in water and the swelling ratio increases with the increasing temperature. Taking into consideration that the HPMC-based hydrogels are prepared under mild reaction conditions with an adjustable gelation time and thermal stability, the method described here has a potential application in biomaterials, especially in the areas of tissue-engineering and drug-controlled release carriers.     

Chlorine atom and ClO wall reaction products

The products of the heterogeneous reactions of chlorine atoms and chlorine oxide radicals with acid coated Pyrex walls have been directly determined for the first time. Contrary to the usual assumption that chlorine atoms recombine to form Cl₂, we find that the major product is HCl, with small amounts of perchlorate also formed. Similarly, ClO radicals form HCl rather than Cl₂. The source of hydrogen for these reactions is probably the water always found in this type of vacuum system. These results may change the interpretation of flow tube experiments with chlorine atoms. Application to the H + HCl reaction is discussed as an example.     

Impact of reaction products on the Grignard reaction with silanes and ketones

Grignard reactions with alkoxysilanes or carbonyl compounds produce alkoxymagnesium halides as by-products. Kinetic measurements for reactions of silanes and of a ketone were performed with Grignard reagents, enriched in alkoxymagnesium halides and taken in a great excess.The alkoxide-type reaction products complex tightly with Grignard reagents and enhance in this way their nucleophilicity, thus accelerating the reaction. However, alkoxides branched at α-C atom exert an unfavorable steric hindrance to reaction resulting in a decrease in the reaction rate.     

α-蒎烯水合主产物的分离与提纯

根据α-蒎烯水合产物中的龙脑、异龙脑和α-松油醇在邻苯二甲酸酐存在下发生酯化、脱水反应的选择性差异，可将三者大部分分离，提高优质龙脑和α-松油醇的得率。．     

Separation of miRNA and its methylation products by capillary electrophoresis

Methylation is a crucial step in plant microRNA biogenesis. To improve our understanding of the methylation process and its regulation, a rapid and convenient high-throughput method should be sought to help with the study of reaction kinetics and assist the search for chemical inhibitors of the methyltransferase, HEN1. In this short communication, we report a pioneering work that used capillary electrophoresis (CE) to separate the miRNA and its methylation product. Capillary zone electrophoresis (CZE) with UV-absorption detection and a reduced running temperature achieved good separation of miR173/miR173* and miR173-m/miR173*-m with a detection limit of around 1 microM. To enhance detection sensitivity, capillary gel electrophoresis (CGE) coupled with laser-induced fluorescence (LIF) detection was also tested, and base-line separation of nanomolar duplex RNA samples was achieved using 4% polyvinylpyrrolidone (PVP) as the sieving matrix and SYBR Green II RNA gel stain for on-column labeling. Although further study is needed to investigate if the separation is sequence dependent, our study demonstrated, for the first time, that CE could be an effective and rapid method in monitoring the miRNA methylation process.