Homochirality and life

Before the emergence of life, left-handed amino acids (L-enantiomers) were selected and right-handed amino acids (D-enantiomers) were eliminated on the primal earth. Nevertheless, with the progress of analytical methods, D-amino acids have recently been found in higher order living organisms in the form of free amino acids, peptides, and proteins. Free D-amino acids have numerous physiological functions. D-amino acids containing animal peptides are well known as opioid peptides. D-amino acids in protein are related to aging. In this review, we describe the D-amino acids that are present and function as D-amino acid biosystems in our bodies.     

Profile and relative concentrations of fatty acids in corn and soybean seeds from transgenic and isogenic crops

In this work 44 fatty acids, which were analyzed as methyl esters by GC/MS in scan mode, have been determined in genetically modified corn and soybean seeds. Their relative concentrations have been compared with those of isogenic lines grown in the same conditions. Studied compounds comprised saturated and unsaturated fatty acids, including cis/trans isomers and minor fatty acids. A classical soxhlet extraction and an accelerated solvent extraction have been assayed to extract the fatty compounds from seeds and the GC separation has been carried out on a biscyanopropylpolysiloxane chromatographic column. Soxhlet extraction was selected as the most convenient and applied to compare the samples. Specific compounds, which could denote the origin of the crop have not been observed, but for some sample pairs, significant differences have been found in relation to the percentage of certain acids; the highest differences for major acids were 4.1% in corn and 4.8% in soybean. The concentrations of long chain acids such as 24:0, 26:0 and 28:0 were higher in some isogenic lines whereas the concentrations of short chain acids such as 6:0, 8:0, 9:0, 10:0 and 12:0 were higher in their transgenic counterparts.     

Simultaneous quantitation of Krebs cycle and related acids by mass fragmentography

Methods are described for simultaneous quantitation of Krebs cycle and related acids by gas chromatography--mass spectrometry using deuterium-labelled acids and n-butyl-d9-esters of the organic acids as internal standards. Using sulphuric acid as esterification catalyst, only lactic, succinic, fumaric, malic, maleic and citric acids were found to be stable to hydrogen exchange and could be used as reference standards in the deuterated form. In contrast, pyruvic, oxalacetic, alpha-ketoglutaric and malonic acids were found to exchange their deuterium readily and could not be employed for this purpose. All the acids could be quantitated using n-butyl-d9-esters of reference organic acids as internal standards, following a separate preparation of the n-butyl derivatives of the unknown acids. The method is suitable for routine analysis of organic acids at the picogram level in perchloric acid extracts of tissues.     

Capillary electrophoresis determinative and GC-MS confirmatory method for water-soluble organic acids in airborne particulate matter and vehicle emission

Urban fine airborne particulate matter (PM2.5) and vehicle emission samples were studied for water-soluble low-molecular-weight carboxylic acids using CE with indirect UV detection. Further identification of these acids was achieved using GC-MS as their butyl esters (after derivatization with BF3/butanol). Several dicarboxylic acids in the range C2-C10 including straight-chain, branched-chain, cis- and trans-unsaturated, and aromatic acids were confirmed by GC-MS. In addition, aromatic acids such as benzoate, phthalate, terephthalate, isophthalate, and 4-methylphtalate were present in such samples, but some of these were not well resolved by the used CE method. Oxocarboxylic acids (Cn(w) with n > 4) were also identified by GC-MS but not determined by CE due to lack of standards. The rapidity and simplicity of the CE method were clearly demonstrated, and the method was observed to be advantageous for routine monitoring of water-soluble organic acids in airborne PM2.5 and vehicle emission at low microg/L levels.     

Simultaneous Quantification of Organic Acids in Tamarillo (Solanum betaceum) and Untargeted Chemotyping Using Methyl Chloroformate Derivatisation and GC-MS

Sixteen organic acids were quantified in peel and pulp of Amber, Laird’s Large and Mulligan cultivars of tamarillo using GC-MS. Fourteen of these compounds had not previously been quantified in tamarillo. An untargeted metabolomics approach was used in parallel to identify and quantify 64 more metabolites relative to the internal standard, indicating abundances of glutamic acid, pro-line, aspartic acid and γ-aminobutyric acid as well as lower concentrations of several other essential fatty acids and amino acids. The main findings were that total organic acid concentration was significantly higher (p < 0.05) in pulp than in peel, with the highest concentration seen in Mulligan pulp (219.7 mg/g DW). Remarkably, after citric acid, the potent bactericide itaconic acid was the second most abundant organic acid. At least 95% of organic acids in tamarillo were one of these two acids, as well as cis-aconitic, malic and 4-toluic acids. Differences between cultivar chemotypes were as substantial as differences between tissues. These results suggest that the bitter flavour of the peel does not result from organic acids. The combination of targeted and untargeted metabolomics techniques for simultaneous qualitative and quantitative investigation of nutrients and flavours is efficient and informative.     

NMR metabolic profiling of organic and aqueous sea bass extracts: implications in the discrimination of wild and cultured sea bass

The nuclear magnetic resonance (NMR) technique was used as analytical tool to determine the complete metabolic profiling of sea bass extracts: water-soluble metabolites belonging to different classes such as sugars, amino acids, dipeptides and organic acids as well as metabolites soluble in organic solvent such as lipids, sterols and fatty acids were identified. The metabolite profiling together with a suitable statistical analysis were used to discriminate between wild and cultured sea bass samples. Preliminary results show that discrimination between wild and cultured sea bass was obtained not only using fatty acid composition but also cholesterol and phosphatidylethanolamine and some water-soluble metabolites such as choline, trimethylamine oxide, glutamine, fumaric and malic acids.     

Comparative characteristic studies on soil and commercial humic acids

Summary Two commercial and eight soil humic acids were characterized to evaluate the use of commercial humic materials in soil and environmental studies. Significant differences between commercial and soil humic acids were observed in ash content, major and micro-element constituents, E₄/E₆ ratio, total acidity and ¹³C-NMR spectroscopy; but data from stability constant and IR-spectroscopy were almost identical. The commercial materials appeared to be significantly less aromatic and more aliphatic in nature as compared with representative soil humic acids. The above discrepancies were interpreted as indicating an origin of the respective commercial product other than soil. The two commercial humic acids investigated may not originate from the same source material as evidenced by clear-cut differences in the intensity of the ¹³C-NMR spectra in almost all chemical shift regions. Due to the observed structural, compositional and characteristic dissimilarities, both between soil and commercial humic acids and between different commercial ones, the commercial humic products are not recommended to be used as analogues of real soil humic acids in soil and environmental studies.     

卤代苯丙氨酸衍生物的合成与拆分

以苯胺类化合物为起始原料,经重氮化、丙烯酸加成、氨解等过程合成了5种消旋苯环取代苯丙氨酸衍生物,并采用α-糜蛋白酶和枯草杆菌蛋白酶对所有氨基酸进行了拆分,通过IR,¹HNMR,元素分析和旋光等技术进行表征.其中4种氨基酸未见文献报道.     

Identification of very-long-chain fatty acids in rat and mouse harderian gland lipids by capillary gas chromatography-mass spectrometry

Lipids of Harderian ophthalmic gland were separated by means of thin-layer chromatography with flame ionization detection in an latroscan apparatus. Wax ester and polar lipids (phosphatidylethanolamine and phosphatidylcholine) were detected as the main lipids in rats and glyceryl ether diester and both polar lipids were the main lipids in mice. Fatty acids were determined in individual lipid classes by means of gas chromatography and gas chromatography-mass spectrometry on capillary columns. The content of fatty acids, the positional isomers of monoenoic acids being predominantly C18, C20 and C22, is most interesting. Very-long-chain fatty acids, saturated fatty acids up to C30 and even monoenoic acids up to C28 were detected. Branched-chain fatty acids, predominantly iso and anteiso, are minority components, although their chain length distribution (C15-C27) is broad.     

Fatty acids and amino acids of entomopathogenic fungus <Emphasis Type="Italic">Conidiobolus coronatus</Emphasis> grow on minimal and rich media

Entomopathogenic fungi are referred to as potential candidates as insect pest control agents. The objective of the study was to identify fatty acids and amino acids from Conidiobolus coronatus cultured on two different media. Each medium was extracted with ethyl acetate and its mixtures with isopropanol, acetonitrile and methanol. Analyses of fatty acids and amino acids of entomopathogenic fungus C. coronatus were performed by means of gas chromatography coupled with mass spectrometry. The analysis showed that the fungus C. coronatus produces the following groups of compounds: fatty acids and amino acids; α- and β-glucopyranose were also identified. The identified fatty acids included 12–20, 22 and 24 carbon atoms per chain. The highest content of fatty acids was detected in a mycelium sample cultured in a liquid minimal medium extracted with ethyl acetate. The lowest content of these organic compounds was identified in mycelium cultured in a liquid nutrient-rich medium extracted with ethyl acetate–methanol mixture. Fatty acids were found to account for 62.0 mass % to 94.4 mass % of all organic compounds in the analyzed mycelia. C18:1 acids were detected in the highest amounts when ethyl acetate was used as the extracting agent. The identified amino acids accounted for 4 mass % to 21 mass % of all organic compounds. Upon extraction of C. coronatus mycelium samples with the ethyl acetate—methanol mixture, two anomeric forms of glucose were also identified. An analysis of the studied material confirmed, that the entomopathogenic fungus C. coronatus is a very rich source of organic compounds, which might encourage its further research so as to identify an even larger number of compounds being produced by this species.     

Bile acids and sterols in urban sewage treatment plants

The composition of bile acids, sterols and sterones in water and sludge from an urban sewage treatment plant has been examined for assessment of the possible use of these compounds as pollution biomarkers. Samples were solvent-extracted, hydrolysed, and fractionated by column chromatography to separate acids, hydrocarbons, sterones and sterols. These fractions, except hydrocarbons, were methylated (acids only) and silylated for instrumental analysis. Gas chromatographic-mass spectrometric analysis was performed in the electron-impact mode, using a non-polar capillary column. Lithocholic acids (3alpha- and 3beta-epimers), coprostanone, coprostanol, cholesterol, cholestenone, and cholestanone were found in sludge and all waters. However, the waters after secondary plant treatment contained mainly lithocholic acids epimers and coprostanone, pointing to these compounds as potential markers for urban treatment plant effluents in natural waters courses.     

Investigation of the possible role of fatty acids and alcohols as precursors of normal hydrocarbons generated during vacuum pyrolysis of a young kerogen

Significant amounts of fatty acids and alcohols, in addition to straight-chain hydrocarbons, were observed in the vacuum pyrolysate of a young kerogen isolated from a fresh-water sediment. The amounts and the distribution patterns of fatty acids and alcohols were compared with those indigenous to the original kerogen, as determined by solvent extraction followed by saponification extraction. Straight-chain fatty acids in the pyrolysate were 12.7 mg/g kerogen, while those extracted from the original kerogen were 10.5 mg/g. The amounts of fatty alcohols in the pyrolysate and in the extracts were 1.31 and 1.10 mg/g. respectively. The excess amounts in the pyrolysate were explained by the release of a strictly bound form of fatty acids and alcohols, which were left in the kerogen matrix even after the saponification extraction. The results indicated that most fatty acids and alcohols were not decomposed as a result of the vacuum pyrolysis and that neither fatty acids nor fatty alcohols should be viewed as the prime precursors of the straigth-chain hydrocarbons (4.6 mg/g) simultaneously generated during vacuum pyrolysis.     

Chiral lariat ethers as membrane carriers for chiral amino acids and their sodium and potassium salts

Four chiral lariat ethers 8–11 containing a (p-methoxyphenoxy) methyl side arm were used for chiral discrimination of amino acids in their zwitterionic form or as potassium and sodium salts in transport across a bulk chloroform membrane with satisfactory selectivity. The carriers that were employed exhibited different transport selectivity relative to the amino acids and their salts under study. The d/l selectivity strongly depends on the amino acids or their salts, and in some cases reverse selectivity has been obtained. The best selectivity was obtained in the case of tyrosine and its potassium salts for all carriers. The transport rates of amino acids and their salts were found to be controlled by factors such as the structure of the carriers and amino acids or their salts. Among these factors, it was also found that the side arm of the lariat ethers plays an important role in the transport process. As a consequence, the main goal of our investigation was to separate the chiral amino acids through liquid membranes.     

An improved method for preparation of perbenzoylated ganglioside-derived sialic acids and nanogram detection of N-acetyl- and N-glycolylneuraminic acid by high performance liquid chromatography.

An improved method for the preparation of perbenzoylated ganglioside-derived sialic acids is described. After mild acid hydrolysis, isolation of sialic acids can be achieved by Folch partition (Method A) or by anion exchange chromatography (Method B). Perbenzoylated sialic acids were freed from benzoylation reagents by a second Folch partition. Total recoveries of both methods were found to be greater than or equal to 90%, calculated from metabolically labelled gangliosides. Derivatized N-acetylneuraminic and N-glycolylneuraminic acids were separated and quantified by isocratic high performance liquid chromatography using a RP18 column as the stationary phase and methanol:water (8:2) as the mobile phase. Both sialic acids were completely separated and eluted as single peaks within 15 min, monitored by UV detection. As little as 20 ng of neuraminic acid could be detected, the detector being linear up to 5 micrograms tested.     

Electron ionization mass spectra and their reproducibility for trialkylsilylated derivatives of organic acids, sugars and alcohols

Mass spectra of trialkylsilyl derivatives of fatty acids, dicarboxylic acids, hydroxyacids, oxoacids, sugars, amino acids and alcohols were obtained. Amino acids were analyzed as tert-butyldimethylsilyl derivatives; all other model compounds were analyzed as trimethylsilyl derivatives. Reproducibility of the electron ionization (EI) mass spectra for the derivatives obtained was discussed. It was shown that, for many investigated derivatives, composition of the respective mass spectra depended greatly on ion source contamination. The trimethylsilylated alpha-tocopherol mass spectrum composition was most significantly influenced by ion source contamination. This compound can be used to test ion source contamination.     

GC Analysis of Organic Acids and Phenols Using On-Line Methylation with Trimethylsulfonium Hydroxide and PTV Solvent Split Large Volume Injection

The combination of on-line methylation using trimethylsulfonium hydroxide with large volume injection of 100 μL was evaluated for the analysis of organic acids and phenols in water. Solvent split injection was applied with complete evaporation of the solvent before analytes were transferred onto the GC column. Despite complete solvent removal, losses were very low compared to conventional splitless injection even for volatile acidic compounds such as propionic acid and phenol. This is explained by intermediate formation of low volatility trimethylsulfonium salts of the analytes which were held in the injector for long evaporation times of up to 10 min, if the evaporation temperature was as low as 10°C. Using a simple liquid/liquid extraction procedure, volatile fatty acids, dicarboxylic acids, benzoic acids and phenols could be detected in 5 mL of water at concentrations of 0.04–0.1 μmol/L with GC/MS in full scan mode. Lactic, pyruvic, and also malonic acids could only be detected at higher levels because of their limited extractability from water as well as their poorer methylation yields. The method provides an easy way to sensitively detect acidic compounds of medium to high volatility in water. It was applied for screening of organic acids and phenols in batch cultures of anaerobic bacteria of which one example is shown.     

Boc-L-甲基苯丙氨酸的合成与拆分

利用Boc-2-氨基丙二酸二乙酯和甲基苄溴为原料, 合成邻位、间位、对位甲基取代的Boc-苯丙氨酸乙酯, 经枯草杆菌蛋白酶拆分得到对应的Boc-L-甲基苯丙氨酸. 通过红外光谱、核磁共振、质谱及旋光度分析对3种物质的结构进行了表征.     

Homopolymerization of Four New Hydroxyamic Acids and Polycondensation of Glycerol With Dibasic Amic Acids

During the past 15 years or so, research in our laboratory has been mainly directed toward the synthesis of cyclic imides and isoimides for different purposes [1–4]. Thus a large number of substituted amic acids and bis-amic acids were prepared as intermediates. These amic acids were readily purified, crystallized, and obtained in excellent yields. In the present paper two types of amic acids are prepared. The first type is hydroxyamic acids of formula A, while the second type consisted of dibasic amic acids such as B:     

Ascertainment of D-amino acids in germ-free, gnotobiotic and normal laboratory rats.

Free D-amino acids were ascertained in the blood serum, urine and aqueous ethanolic extracts of feces of germ-free laboratory rats and a rat made gnotobiotic (tetra-associated) with species of Streptococcus, Lactobacillus and Clostridium. D-Amino acids were also determined in the brains of two germ-free rats. For comparison, D-amino acids were also measured in the blood serum of normal rats and the blood plasma, urine and feces of normal white mice. D-Enantiomers of most protein L-amino acids were detected in all physiological samples of animals. Quantities of free D-amino acids were determined as N(O)-pentafluoropropionyl-(2)-propyl esters by enantioselective gas chromatography and mass spectrometry. Stereoisomers of the bacterial marker 2,6-diaminopimelic acid, analyzed as N-trifluoroacetyl-(2)-propyl esters, were detected in feces of the gnotobiotic but not of the germ-free rat.     

Determination of 4-aminobutyric acid, aspartate, glutamate and glutamine and their 13C stable-isotopic enrichment in brain tissue by gas chromatography-mass spectrometry

A selected-ion monitoring method was developed for measuring 4-aminobutyric acid, aspartate, glutamate, and glutamine in brain tissue. Natural isotopes of these amino acids and their stable-isotopic enrichment following intravenous infusion of a precursor, [13C]glucose, were quantitated. Frozen mouse brain tissue was homogenized in cold 80% ethanol, and the supernatant, equivalent to 1 mg of wet weight brain tissue, was extracted using solid-phase bonded silica ion-exchange columns. Aspartate and glutamate (dicarboxylic acids) were isolated from strong anion-exchange columns, whereas 4-aminobutyric acid and glutamine (neutral amino acids) were isolated from strong-cation exchange columns. n-Butyl ester pentafluoropropionyl amide derivatives of these amino acids were analyzed by gas chromatography-mass spectrometry using a methane positive chemical ionization mode after gas chromatographic separation on a wide-bore, fused-silica capillary column. The method is applicable to determination of brain concentrations of these amino acids as well as their fluxes following administration of a stable-isotopic tracer.