Elucidation of the structures of residual and dissolved pine kraft lignins using an HMQC NMR technique

Comparative studies on the structures of residual and dissolved lignins isolated from pine kraft pulp and pulping liquor have been undertaken using the (1)H-(13)C HMQC NMR technique, GPC, and sugar analysis to elucidate the reaction mechanisms in kraft pulping and the lignin reactivity. A modified procedure for the isolation of enzymatic residual lignins has resulted in an appreciable decrease in protein contaminants in the residual lignin preparations (N content < 0.2%). The very high dispersion of HMQC spectra allows identification of different lignin moieties, which signals appear overlapped in 1D (13)C NMR spectra. Elucidation of the role of condensation reactions indicates that an increase in the degree of lignin condensation during pulping results from accumulation of original condensed lignin moieties rather than from the formation of new alkyl-aryl structures. Among aryl-vinyl type moieties, only stilbene structures are accumulated in lignin in appreciable amounts. Benzyl ether lignin-carbohydrate bonds involving primary hydroxyl groups of carbohydrates have been detected in residual and dissolved lignin preparations. Structures of the alpha-hydroxyacid type have been postulated to be among the important lignin degradation products in kraft pulping. The effect of the isolation method on the lignin structure and differences between the residual and dissolved lignins are discussed.     

pH Dependence of the progression in NMR T(2) relaxation times in post-mortem muscle

Continuous NMR T(2) relaxation measurements were carried out on seven rabbit longissimus muscle samples in the period from 25 min to 28 h post-mortem at 200 MHz for (1)H. To display differences in post-mortem pH progress and extent of changes in water characteristics during conversion of muscle to meat, three of the seven animals were pre-slaughter injected with adrenaline (0.5 mg/kg live weight 4 h before sacrifice) to differentiate muscle glycogen stores at the time of slaughter. Distributed analysis of T(2) data displayed clear differences in the characteristics of the various transverse relaxation components dependent on progress in pH, as did the water-holding capacity of samples 24 h post-mortem. This reveals a pronounced effect of the progressive change in pH on the subsequent development in physical/chemical states of water during the conversion of muscle to meat. Finally, the relaxation characteristics are discussed in relation to supposed post-mortem processes of protein denaturation.     

Application of the membrane interphase probe (MIP): an evaluation

Background, aim, and scope

The membrane interphase probe (MIP?) from Geoprobe Systems® has frequently been applied in different countries for the characterization of soil contaminated with volatile organic carbons (VOCs). Experience shows that misinterpretation of the collected data is common. This is mainly due to the lack of understanding and knowledge related to the detectors used, their detection limits, and the sensitivity of the MIP system. It has been noticed that the sensitivity of the system given by the producer and by different users are rather optimistic, e.g., the values given are lower (= better) than those actually experienced in the field. A need for a better understanding of the MIP system sensitivity, combined with a more scientifically based interpretation of the collected data, exists.

Materials and method

Both laboratory tests (using solutions) as well as field measurements were carried out using different detector configurations to allow a better interpretation of the detector signals/system sensitivity and to collect qualitative information. These configurations were: (1) detectors stand alone; (2) the use of a 2-ml sample loop, and (3) a purge and trap system. The configurations (2) and (3) are used in combination with a capillary column to carry out on-site qualitative and semiquantitative analyses.

Results and discussion

With respect to the configuration of “detectors stand alone,” detection limits for toluene (in aqueous solutions) range between 4 ppm (flame ionization detector—FID) and 10 ppm [photo ionization detector (PID)]. For chlorinated aliphatic hydrocarbons (CAHs), observed limits are 10 ppm (FID), 4–50 ppm (PID), and 3–10 ppm [dry electrolytic conductivity detector (DELCD)]. When using the 2-ml sample loop, relatively high concentrations have to be initially present in the soil. Observed detection limits for mono-aromatic hydrocarbons are 5–100 ppm; for CAHs, 1–50 ppm; for alkanes, 250–400 ppm; and for MTBE, 25 ppm. The application of purge and trap results in a better resolution and the detection of lower concentrations. Consequently, a better identification of the pollution with depth is possible. In this case, the detection limits are a function of the concentrations and the flushing time. In relation to the qualitative analyses, it was found that the configuration of the MIP-system with the built-in capillary column and the 2-ml sample loop or the purge and trap preconcentrator, respectively, are useful to carry out on-site analyses, thus allowing a better identification of the pollution in a vertical profile.

Conclusions

The measurements carried out using the MIP with detectors stand-alone or in combination with a loop or trap, or connected to a column, confirm that analysis is indeed very useful to characterize VOC source zones when knowing and understanding its performance. This relates mainly to the detection limits of the MIP system. For a selection of parameters, such limits have been obtained. These values seem to be more realistic than those found in the few references where numbers are given. For the qualitative measurements, it can be concluded that a better resolution is obtained, and pollutants present in lower concentrations will be detected when using the purge and trap. It is advised to determine the optimal flushing time and the detection limit of the expected pollutants in advance.

Recommendations and perspectives

This study indicates that there is still a need for further measurements and discussion between users. Finally, additional data should result in a better interpretation of the collected field data.     

High-field nuclear magnetic resonance (NMR) study of truffles (Tuber aestivum vittadini)

A high-field NMR technique was used to analyze aqueous and organic extracts of truffles (Tuber aestivum vittadini) to characterize their chemical composition. Water-soluble metabolites belonging to different classes such as sugars, polyols, amino acids, and organic acids were almost completely assigned by means of one- and two-dimensional experiments (1H-1H COSY, TOCSY, 1H-13C HSQC, 1H-13C HMBC, and 1H-31P HMBC). The 1H spectral assignment of the cell membrane components such as lipids, sterols, and fatty acids extracted in organic solvents was also performed.     

Biosynthesis, molecular structure, and domain architecture of potato suberin: a (13)C NMR study using isotopically labeled precursors

Although suberin in potato wound periderm is known to be a polyester containing long-chain fatty acids and phenolics embedded within the cell wall, many aspects of its molecular structure and polymer-polymer connectivities remain elusive. The present work combines biosynthetic incorporation of site-specifically (13)C-enriched acetates and phenylalanines with one- and two-dimensional solid-state (13)C NMR spectroscopic methods to monitor the developing suberin polymer. Exogenous acetate is found to be incorporated preferentially at the carboxyl end of the aliphatic carbon chains, suggesting addition during the later elongation steps of fatty acid synthesis. Carboxyl-labeled phenylalanine precursors provide evidence for the concurrent development of phenolic esters and of monolignols typical of lignin. Experiments with ring-labeled phenylalanine precursors demonstrate a predominance of sinapyl and guaiacyl structures among suberin's phenolic moieties. Finally, the analysis of spin-exchange (solid-state NOESY) NMR experiments in ring-labeled suberin indicates distances of no more than 0.5 nm between pairs of phenolic and oxymethine carbons, which are attributed to the aromatic-aliphatic polyester and the cell wall polysaccharide matrix, respectively. These results offer direct and detailed molecular information regarding the insoluble intermediates of suberin biosynthesis, indicate probable covalent linkages between moieties of its polyester and polysaccharide domains, and yield a clearer overall picture of this agriculturally important protective material.     

Water distribution in brine salted cod (Gadus morhua) and salmon (Salmo salar): a low-field 1H NMR study

Low-field (LF) (1)H NMR T 2 relaxation measurements were used to study changes in water distribution in lean (Atlantic cod) and fatty (Atlantic salmon) fish during salting in 15% NaCl and 25% NaCl brines. The NMR data were treated by PCA, continuous distribution analysis, and biexponential fitting and compared with physicochemical data. Two main water pools were observed in unsalted fish, T 21, with relaxation times in the range 20-100 ms, and T 22, with relaxation times in the range 100-300 ms. Pronounced changes in T 2 relaxation data were observed during salting, revealing changes in the water properties. Salting in 15% brine lead to a shift toward longer relaxation times, reflecting increased water mobility, whereas, salting in saturated brines had the opposite effect. Water mobility changes were observed earlier in the salting process for cod compared to salmon. Good linear correlations ( F 相似文献   

Direct electron paramagnetic resonance study of tobacco. 1. Manganese(ii) as a marker

Three categories of tobacco products were studied using electron paramagnetic resonance (EPR) spectroscopy: Cuban cigar brand name Montecristo, four international trademark cigarettes, and three types of Middle Eastern tobacco blends called Al-Moassal or Jurak. The Montecristo Cuban cigar is used as standard of high-quality tobacco. Mainly two EPR signals from all of the studied samples are observed: a very weak sharp EPR signal superimposed on a broad signal. The broad EPR signal is attributed to a manganese(II) complex. The intensity of the manganese(II) EPR signal is found to be related to the quality of the tobacco content. The sharp signal, which is characteristic of semiquinone radicals, is observed at room temperature, and its intensity increases drastically with temperature.     

(13)C NMR characterization of triacylglycerols of Moringa oleifera seed oil: an "oleic-vaccenic acid" oil

The composition of acyl chains and their positions in the triacylglycerols of the oil extracted from seeds of Moringa oleifera were studied by (13)C NMR spectroscopy. The unsaturated chains of M. oleifera seed oil were found to comprise only mono-unsaturated fatty acids and, in particular, two omega-9 mono-unsaturated acids, (cis-9-octadecenoic (oleic acid) and cis-11-eicosenoic acids) and one omega-7 mono-unsaturated acid (cis-11-octadecenoic acid (vaccenic acid)). The mono-unsaturated fatty acids were detected as separated resonances in the spectral regions where the carbonyl and olefinic carbons resonate according to the 1,3- and 2-positions on the glycerol backbone. The unambiguous detection of vaccenic acid was also achieved through the resonance of the omega-3 carbon. The (13)C NMR methodology enabled the simultaneous detection of oleate, vaccenate, and eicosenoate chains according to their positions on the glycerol backbone (1,3- and 2-positions) through the carboxyl, olefinic, and methylene envelope carbons of the triacylglycerol acyl chains.     

Geographical characterization of italian extra virgin olive oils using high-field (1)H NMR spectroscopy.

1H high-field nuclear magnetic resonance (NMR) was used to analyze 216 extra virgin olive oils collected in three years (1996, 1997, and 1998) in different Italian areas in order to evaluate the potential contribution of this technique to the geographical characterization of olive oils. A statistical procedure performed on the intensity of selected NMR peaks has been proposed. Tree clustering analysis of NMR data performed without any a priori hypothesis showed the existence of reliable parameters able to group the olive oils according to the location of olive oil production. Linear discriminant analysis applied to selected NMR parameters of olive oils of the same year of production allowed the grouping of samples according to their geographical origin with only very few errors. Moreover, a satisfactory grouping is reached by combining the NMR data of olive oils from two different years (1996 and 1997). Operating on appropriate sampling, a careful analysis of data yielded the conclusion that the place of olive production could be singled out as a discriminating factor regardless of the cultivars from which the olive oils are derived.     

Evaluation of forage maize yield gap using an integrated crop simulation model-satellite imagery method (Case study: four watershed basins in Golestan Province)

The yield gap provides a guide for prioritization of crop management options (such as optimized sowing date, seeding rate, irrigation schedule, soil fertility, fertilizer application, weed and pest control) in a studied area. This study aimed to determine the maize yield gap in four major watersheds in Golestan Province, Iran, using an integrated crop simulation model-satellite imagery method. The actual yield estimated by the NDVI (as the selected index) was between 8.89 and 20.40 t ha^?1, while the potential yield is between 19.03 and 22.35 t ha^?1. About 91.76% of the studied area had a yield less than 85% of the potential yield. The lowest actual yield was in the south, southeast, and north of the study area. The yield gap was estimated between 0 to 11.76 t ha^?1 and 66.66% of maize farms yield gap was between 3.5 to 5.5 t ha^?1. The yield gap fraction changed between 0 and 0.57. The results showed that soil-dependent variables, slope, and fluctuation in farm management factors (plant density, planting, irrigation, and various methods of weed control) caused the yield gap. High yield gap indicates that there is an opportunity to increase production through managerial optimization or excluding maize from cropping patterns.     

Metabolite profiling using (1)H NMR spectroscopy for quality assessment of green tea, Camellia sinensis (L.)

A set of 191 green teas from different countries was collected and analyzed by (1)H NMR. It was proposed to establish if the teas could be discriminated according to the country of origin or with respect to quality. Both principal component analysis (PCA) and cluster analysis were applied to the data. Some separation of Chinese and non-Chinese teas was observed. The present results did not allow allocation of samples to individual countries, but cluster analysis suggested that it might be possible with an augmented sample set. The PCA did show a separation between the Longjing type (highest quality Chinese tea) and most other Chinese teas and indicated some metabolites that could be responsible for the difference. Longjing teas showed higher levels of theanine, gallic acid, caffeine, epigallocatechin gallate, and epicatechin gallate and lower levels of epigallocatechin when compared with other teas. These compounds have been mentioned previously in connection with quality, but it was also shown that higher levels of theogallin (5-galloyl quinic acid), theobromine, 2-O-(beta-l-arabinopyranosyl)-myo-inositol and some minor sugar-containing compounds were found in Longjing teas while higher levels of fatty acids and sucrose were found in the other teas. These new markers could prove to be useful for the authentication of bulk tea.     

Polychlorinated biphenyls (PCBs) detection in food samples using an electrochemical immunosensor

In this work, a disposable electrochemical immunosensor, based on a competitive assay scheme, was applied to detect polychlorinated biphenyls (PCBs) in food. For this purpose, antibodies against PCBs were directly immobilized onto the carbon surface of a disposable screen-printed electrode. A competition between the PCBs present in the sample and a fixed concentration of an enzyme-labeled PCB was realized and evaluated by electrochemical detection. Alkaline phosphatase was used as the enzyme label, coupled with differential pulse voltammetry (DPV) as the electrochemical technique. The immunosensor was tested on aroclor mixture detection (1242 and 1248) and then on some typologies of food samples to evaluate the possible application for real sample analysis. Samples analyzed were from different matrixes, such as sheep milk, bovine adipose tissue, and bovine muscle. Results obtained were compared with the accredited results according to ISO 17025 methods for PCB detection (HRGC-LRMS) as a confirmatory analysis. Preliminary results show the possibility to use this device as a screening method in food sample analysis. The negligible matrix effect observed may lead to a simplified extraction procedure, and considerable time and consumable savings are the immediate benefits given by the proposed method.     

Geographical extrapolation of forest soil acidification using pH(KCl) data: A case study

Soil acidification over the last three decades in three forest areas situated in northern Belgium (Region of Flanders), is assessed by comparing recent data of soil analysis with historical data. It is investigated whether acidification values of sites evenly spread over the region, averaged per soil group, can be used to assess acidification in specific forest areas. Results suggest that data obtained by this kind of geographical extrapolation do not differ significantly from measured data. In sandy soils however, a stop of acidification as measured by the pH(KCl), is observed at pH(KCl) values lower than 3.3.     

A 13C NMR study of decomposing logging residues in an Australian hoop pine plantation

Purpose

Residue retention is important for nutrient and water economy in subtropical plantation forests. We examined decomposing hoop pine (Araucaria cunninghamii Ait. Ex D. Don) residues—foliage, branches, and stem wood—to determine the changes in structural chemistry that occur during decomposition.

Materials and methods

Residues were incubated in situ using 0.05 m² microplots. We used solid-state ¹³C nuclear magnetic resonance (NMR) spectroscopy to determine the structural composition of harvest residues in the first 24 months of decomposition.

Results and discussion

The spectral data for branch and stem residues were generally similar to one another and showed few changes during decomposition. The lignin content of branch and foliage residues decreased during decomposition. When residues were mixed together during decomposition, the O-alkyl fraction of foliage decreased initially then increased up to 24 months, while the alkyl carbon (C) fraction exhibited the opposite pattern. The decomposition of woody hoop pine residues (branch and stem wood) is surprisingly uniform across the major C forms elucidated with ¹³C NMR, with little evidence of preferential decomposition. When mixed with branch and stem materials, foliage residues showed significant short- and long-term compositional changes. This synergistic effect may be due to the C/N ratio of the treatments and the structure of the microbial decomposer community.

Conclusions

Twenty-four months of decomposition of hoop pine residues did not result in substantial accumulation of recalcitrant C forms, suggesting that they may not contribute to long-term C sequestration.     

Rheological and nuclear magnetic resonance (NMR) study of the hydration and heating of undeveloped wheat doughs

The undeveloped doughs of two wheat flours differing in technological performance were characterized at the supramolecular level, by fundamental small-deformation oscillatory rheology and shear viscometry, and at the molecular level, by nuclear magnetic resonance (NMR) spectroscopy. For the harder variety, the higher storage moduli indicated lower mobility of the protein/water matrix in the 0.001-100 s range. Conversely, 1H NMR indicated higher molecular mobility in the sub-microsecond range for protein/water, whereas starch was found to be generally more hindered. It is suggested that faster protein/water motions are at the basis of the higher structural rearrangement indicated by tan delta for the harder variety. Rheological effects of heating-cooling reflect mainly starch behavior, whereas 1H NMR spectra and relaxation times give additional information on component mixing and molecular mobility. The heated softer variety dough formed a rigid lattice and, although a similar tendency was seen for the hard variety, all of its components remained more mobile. About 60% of starch crystallizes in both varieties, which may explain their similar rheological behaviors upon cooling.     

Quantitative analysis of limonin in grapefruit juice using an enzyme-linked immunoassay: interlaboratory study

Eleven laboratories participated in an interlaboratory study of an enzyme-linked immunoassay for limonin in grapefruit juice. Participating laboratories received training and practice samples until familiar with the procedure. Laboratories then received 8 sample pairs of grapefruit juice as blind duplicates. Samples were refrigerated and analyzed within 4 days of receipt. Results from 1 laboratory and 6 individual samples were excluded from the statistical analysis. Method performance was measured by the one-way component of variance analysis. Repeatability relative standard deviations (RSDr) ranged from 7.0 to 15.6%. Reproducibility relative standard deviations (RSDR) ranged from 12.9 to 29.4%.     

Origin of the high-frequency resonances in 1H NMR spectra of muscle tissue: an in vitro slow magic-angle spinning study

High-resolution slow magic-angle spinning (150 Hz) 1H PASS NMR spectroscopy is performed on intact excised rat m. tibialis anterior. Untreated muscles and muscles in vitro incubated in Krebs-Ringers buffer based on deuterium oxide are investigated. In the high-frequency region of the 1H NMR spectra, resonances from H4 (approximately 7.1-7.2 ppm) and H2 (approximately 8.2-8.5 ppm) in histidine are observed. In addition, a resonance appears at 6.7 ppm for the untreated muscles. However, this resonance is absent in muscles following incubation in deuterium oxide. On the basis of its behavior in deuterium oxide combined with supplementary measurements for creatine solutions, the 6.7 ppm resonance is ascribed to the amino protons in creatine. Moreover, the present study demonstrates that the observation of the 6.7 ppm resonance depends on pH, which explains earlier reports stating its occasional appearance. Finally, measurements on solutions of ATP/AMP and histidine indicate that both ATP/AMP and histidine contribute to the resonances at approximately 8.2-8.5 ppm in the 1H NMR spectra of muscle tissue.     

Detection of allergenic ingredients using real-time PCR: a case study on hazelnut (Corylus avellena) and soy (Glycine max)

Compliance with the European allergen labeling legislation (Directive 2007/68/EC) is only possible when coupled with appropriate methods to detect allergens in food. The aim of the current study was to develop new real-time PCR assays for the detection of hazelnut and soy and evaluate these assays via comparison with commercially available kits. Although the new assays were not as sensitive as the commercial qualitative assays, they proved to be more specific. Moreover, the cross-reactivity study indicated contamination of some of the food products used with either hazelnut or soy, which presents a risk for the allergic consumer. The assays were able to quantify as few as 5-15 genome copies. This unit, used to express analytical results for allergen detection by means of PCR, needs to be converted to a unit expressing the amount of allergenic ingredient in order to be informative. This study emphasizes that the use of real-time PCR for allergen quantification is complicated by the lack of appropriate reference materials for allergens.     

Study of interactions between food phenolics and aromatic flavors using one- and two-dimensional (1)H NMR spectroscopy

Changes in flavor release and aroma characteristics in a medium including food phenolics may be attributed to an intermolecular interaction between flavor compounds and phenolics. To investigate the interaction, one- and two-dimensional NMR studies have been carried out on the binding of two phenolics, gallic acid and naringin, with three aroma compounds, 2-methylpyrazine, vanillin, and ethyl benzoate. Evaluation of thermodynamic parameters and intermolecular nuclear Overhauser effects reveals that gallic acid can interact more strongly with aromatic flavors than naringin. The supramolecular complexation is also dependent on the structural nature of the flavors, with 2-methylpyrazine and vanillin interacting more strongly than ethyl benzoate. The interaction is principally pi-pi stacking between the galloyl ring and the aromatic ring of the aroma compounds, but secondary hydrogen-bonding effects help to stabilize the complex and enhance the specificity.     

Antioxidant activity in hepatopancreas of the shrimp (Pleoticus muelleri) by electron paramagnetic spin resonance spectrometry

Free radical scavenging properties of hepatopancreas extracts of Pleoticus muelleri were evaluated by electron paramagnetic spin resonance spectrometry methods (EPR) against the stable 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical. The present study was carried out to characterize different physiological stages of the shrimp under environmental and nutritional stress, evaluating the effect on growth, survival, and functional morphology of the hepatopancreas. Feeding trials were carried out on juveniles (1 g initial weight) held in aquaria. Each diet, with different concentrations of vitamins A and E, was tested in triplicate groups during 25 days. The control groups were fed with fresh squid mantle and with a vitamin-free diet. For all of the diets, the extracts exhibited strong DPPH radical scavenging activity, suggesting that the tissue is a powerful natural antioxidant. Individuals fed with different concentrations of vitamin E showed the strongest effect on the DPPH radicals, reducing the DPPH radicals to 50%, after an incubation period of 3 min. In contrast, the extracts of control animals, fed with squid mantle, had the weakest antioxidant activity (4%). These data indicated that the presence of vitamin E in the diet can provide immediate protection against free radicals.