Combined high-field 13C CP MAS NMR and low-field NMR relaxation measurements on post mortem porcine muscles

Changes in postmortem muscle characteristics are investigated in muscles from eight pigs exposed to different combinations of preslaughter stress (exercise on treadmill) and stunning method (CO(2) vs electrical stunning). Solid-state (13)C cross-polarization (CP) magic-angle spinning (MAS) NMR experiments are carried out on a total of 16 rapidly frozen M. longissimus muscle biopsies taken in vivo the day before slaughter and at 45 min postmortem. Simultaneously, low-field NMR T(2) relaxation time measurements are carried out on samples from M. longissimus. Glycogen and lactate are estimated from the (13)C CP MAS spectra, and correlations of r = 0.89 and r = 0.70, respectively, to subsequent biochemical determinations using partial least squares regression (PLSR) are established. Moreover, PLSR reveals that, besides the 72 ppm signal (carbons in glycogen), a signal around 38 ppm, which increases concomitantly with lactate, is also significantly correlated to changes in glycogen/lactate. With the assumption that the 38 ppm signal reflects CH(2) in phosphocreatine/creatine, altered mobility of creatine as a result of dephosphorylation is indicated. Finally, PLSR on the 45 min (13)C CP MAS spectra also reveals correlation (r = 0.54) to the slowest relaxing T(2) population (50 min postmortem), known to reflect extra-myofibrillar water. Subsequently, evaluation of the loading plot in the PLSR analysis reveals that the correlation exclusively is associated to the 52 ppm resonance intensity. With the assumption that this resonance reflects methyl groups in choline/phosphatidyl choline, the intensity changes in the 52 ppm resonance imply alterations in membrane properties. Accordingly, the data indicate a relationship between membrane properties and the amount of water being expelled from muscle cells postmortem, which supports the hypothesis that disruption of membranes is implicated in the postmortem mobilization of muscle water.     

Solid-state selective (13)C excitation and spin diffusion NMR to resolve spatial dimensions in plant cell walls

The average spatial dimensions between major biopolymers within the plant cell wall can be resolved using a solid-state NMR technique referred to as a (13)C cross-polarization (CP) SELDOM (selectively by destruction of magnetization) with a mixing time delay for spin diffusion. Selective excitation of specific aromatic lignin carbons indicates that lignin is in close proximity to hemicellulose followed by amorphous and finally crystalline cellulose. (13)C spin diffusion time constants (T(SD)) were extracted using a two-site spin diffusion theory developed for (13)C nuclei under magic angle spinning (MAS) conditions. These time constants were then used to calculate an average lower-limit spin diffusion length between chemical groups within the plant cell wall. The results on untreated (13)C enriched corn stover stem reveal that the lignin carbons are, on average, located at distances ～0.7-2.0 nm from the carbons in hemicellulose and cellulose, whereas the pretreated material had larger separations.     

Fates of 13C from enriched glucose and glycine in an organic soil determined by solid-state NMR

The transformations of the indigenous ¹³C and the ¹³C from either uniformly enriched ¹³C-D-glucose or ¹³C-glycine added to an organic soil were followed during a 28-day incubation using cross polarization (CP) magic angle spinning (MAS) ¹³C nuclear magnetic resonance (NMR) spectroscopy and dipolar dephased (DDP) MAS ¹³C NMR. The C mineralization was determined from ¹³C remaining by mass spectrometry and from CO₂ evolution by gas chromatography. DDP MAS ¹³C NMR of the unamended soil indicated a transient increase in molecularly mobile ¹³C in the alkyl- and methyl-C over 5 days, which may be due to redistribution of ¹³C in the microbial biomass in response to perturbation. The added glucose-¹³C remaining declined to 43% after 7 days and 34% after 28 days. After 28 days the amount of added glucose-¹³C remaining was 6 times greater than the biomass C at the outset, while the microbial activity (CO₂ production) was 38% greater, indicating that a significant proportion of added glucose-¹³C was not in microorganisms. Added glycine-¹³C declined faster, such that 29% and 8% remained after 7 and 28 days, respectively. After 28 days’ incubation with ¹³C-glucose, the O-alkyl-C, the acetal- and ketal-C, and the methyl- and alkyl-C resonances in CP MAS ¹³C NMR spectra were all enhanced compared with the unamended soil. The calculated T₁ρH values of the O-alkyl-C and the acetal- and ketal-C resonances were less than those of crystalline glucose, indicating that there was no substantial reservoir of unreacted glucose. After 7 days’ incubation with ¹³C-glycine, none of the signals in the CP MAS ¹³C NMR spectra were enhanced when compared with the unamended soil, indicating that the added ¹³C remaining was distributed in undetectable quantities in a range of functionalities. The calculated T₁ρH values indicated that glycine ¹³C was in O-alkyl-C, acetal- and ketal-C and carbonyl-C. T₁ρH values may be more sensitive to changes in the distribution of ¹³C when ¹³C content is low. The DDP MAS ¹³C NMR spectra of both the ¹³C-glucose- and the ¹³C-glycine-amended soil showed that the molecularly mobile alkyl- and methyl-C increased compared with the unamended soil. Received: 27 February 1997     

Solid-State 13C CP/MAS NMR studies on aging of starch in white bread

The effects of storage methods and glycerol on the aging of breadcrumbs were studied using solid-state (13)C CP/MAS NMR. After baking, a shift in C(1) peaks from triplet (A-type) to singlet (V-type) was observed. Addition of glycerol reduced the carbon peak intensities of fresh and aged breads, which correlated well with the DSC amylopectin "melting" enthalpy (r(2) = 0.91). Upon storage of bread with crust in hermetically sealed containers (when moisture migrated from the crumbs to the crust), the (13)C CP/MAS NMR peak intensity increased more rapidly during aging than when the bread was stored without crust. Although addition of glycerol retarded the starch retrogradation, as observed by (13)C CP/MAS NMR and DSC, it accelerated the firming rate. Therefore, bread firming in this case was controlled not only by starch retrogradation but also by other events (such as local dehydration of the matrix or gluten network stiffening).     

Application of P and Al MAS NMR for phosphate speciation studies in soil and aluminium hydroxides: promises and constraints

Magic angle spinning (MAS) nuclear magnetic resonance (NMR) was used to investigate the chemical environment of P in soil and soil components. ³¹P and ²⁷Al MAS NMR spectra are presented of synthetic aluminium hydroxides (amorphous aluminium hydroxide and gibbsite), reacted with P under different conditions of P concentration, temperature and pH. The reaction product is amorphous octahedral aluminium phosphate, which transforms (partly reversibly) to tetrahedral aluminium phosphate upon drying. Results of several experiments on excessively fertilized sandy soil material are discussed. The soil particle fraction smaller than 50 μm was used for NMR analysis. ³¹P and ²⁷Al MAS NMR spectra show a Ca---P pool and an Al---P pool. A six-fold water extraction removes part of both P pools. Oxalate extraction removes all Ca---P and Al---P from the sample. Removal of the labile P pool by the HFO-DMT long-term P desorption technique, does not drastically change the ³¹P MAS NMR spectrum. The formerly mentioned Ca---P and Al---P are thus stable P pools. The ¹H---³¹P cross-polarization (CP) spectrum of the original soil sample revealed a third chemical environment, which was identified as labile Ca---P: this P pool does not appear in the ¹H---³¹P CP spectrum of the soil sample from which the fast P pool had been removed. The combination of MAS and CP MAS NMR can thus reveal at least three different P species in soil, of which two pools were identified as stable, and one as labile.     

Following the decomposition of ryegrass labelled with 13C and 15N in soil by solid-state nuclear magnetic resonance spectroscopy

Investigating the biogeochemistry of plant material decomposition in soil has been restricted by difficulties extracting and identifying organic compounds. In this study the decomposition of ¹³C- and ¹⁵N-labelled Lolium perenne leaves mixed with mineral soil has been investigated over 224 days of incubation under laboratory conditions. Decomposition was followed using short-term rates of CO₂ evolution, the amounts of ¹³C and ¹⁵N remaining were determined by mass spectrometry, and ¹³C and ¹⁵N solid-state nuclear magnetic resonance (NMR) spectroscopy was used to characterize chemically the plant material as it decomposed. After 224 days 48% of the added ¹³C had been lost with a rapid period of C0₂ evolution over the first 56 days. The fraction of cross-polarization magic angle spinning (CP MAS) ¹³C NMR spectra represented by O-alkyl-C signal probably in carbohydrates (chemical shift, 60–90 p.p.m.) declined from 60 to 20% of the spectrum (chemical shift, 0–200 p.p.m.) over 224 days. The rate of decline of the total ¹³C exceeded that of the 60–90 p.p.m. signal during the first 56 days and was similar thereafter. The fraction of the CP MAS ¹³C NMR spectra represented by the alkyl- and methyl-C (chemical shift, 10–45 p.p.m.) signal increased from 5 to 14% over the first 14 days and was 19% after 224 days. CP MAS ¹³C NMR of ¹³C- and ¹⁵N-L. perenne contained in 100-μm aperture mesh bags incubated in the soil for 56 days indicated that the remaining material was mainly carbohydrate but there was an increase in the alkyl- and methyl-C associated with the bag's contents. After 224 days incubation of the labelled ¹³C- and ¹⁵N-L. perenne mixed with the soil, 40% of the added N had been lost. Throughout the incubation there was only one signal centred around 100 p.p.m. detectable in the CP MAS ¹⁵N NMR spectra. This signal corresponded to amide ¹⁵N in peptides and may have been of plant or microbial origin or both. Although there had been substantial interaction between the added ¹⁵N and the soil microorganisms, the associated redistribution of ¹⁵N from plant to microbial tissues occurred within the amide region. The feasibility of following some of the component processes of plant material decomposition in soil using NMR has been demonstrated in this study and evidence that microbial synthesis contributes to the increase in alkyl- and methyl-C content of soil during decomposition has been represented.     

Changes in Rice with Variable Temperature Parboiling: Thermal and Spectroscopic Assessment

Rapid visco analysis (RVA) and differential scannning calorimetry (DSC) provided overall assessments of the effects of variable temperature soaking at 30, 50, 70, and 90°C and steaming at 4, 8, and 12 min. Calculation of the relative parboiling index (RPI) and percent gelatinization provided good metrics for determining the overall effects of partial parboiling. FT‐Raman and solid‐state ¹³C CP‐MAS NMR spectroscopies provided insight to conformational changes in protein and starch of paddy rice under various parboiling conditions. RVA showed lower pasting curves and DSC showed lower ΔH with increased temperature and steaming times. A large decrease in viscosity occurred with only the 30‐4 treatment as opposed to raw rice. This observation was consistent with FT‐Raman results that indicated substantial conversion of the protein from α‐helix to other conformations. DSC indicated incomplete gelatinization of starch, even with 90°C soaking and 12 min of steaming. Solid‐state ¹³C CP‐MAS NMR spectroscopy confirmed this result. However, it indicated the percent of V_h/amorphous plus the remaining crystalline starch in the 90‐12 treatment was equal to the amorphous and partially‐ordered starch in commercially parboiled rice. These results suggest that partial parboiling, 90°C soaking, and more than 8 min of steaming (ideally ≈12 min) of paddy rice is sufficient to induce changes that inactivate enzymes and provide enough starch gelatinization to prevent kernel breakage.     

Solid-state (13)C CP MAS NMR spectroscopy of mushrooms gives directly the ratio between proteins and polysaccharides

The solid-state (13)C CP MAS NMR technique has the potential of monitoring the chemical composition in the solid state of an intact food sample. This property has been utilized to study mushrooms of different species (Pleurotus ostreatus, Pleurotus eryngii, Pleurotus pulmunarius, and Lentinula edodes), already characterized by chemical analyses for protein and dietary fiber components. Solid-state (13)C CP MAS NMR spectroscopy reveals a large difference in the ratio between the glucidic and the proteic resonances probably depending on the mushroom species. An accurate inspection by model compounds and suitable mixtures of proteins and saccharides gives a methodology to interpret these experimental data. A good correlation (R(2) = 0.93; R(2) = 0.81) has been obtained by comparing the NMR data with the results of the chemical analyses. The results suggest the possibility to perform a taxonomic study and/or a nutritional study on the basis of the ratio between protein and polysaccharide levels determined by NMR or chemical methodologies.     

The significance of solid‐state 13C NMR spectroscopy of whole soil in the characterization of humic matter

Abstract

Differences in characteristics of humic matter were investigated by solid‐state CP/MAS ¹³C NMR spectroscopy of whole (nontreated) materials and their extracted humic fractions. Samples used in the analysis were lignite, a commercial humate AG, and the Bh horizons of a Mascotte and a Lawnwood soil. Humic fractions were extracted by the 0.1 M NaOH or Na₄P₂O₇ (pH 9.8) method. The humic (HA) and fulvic acid (FA) obtained were weighed and analysed for total acidity, carboxyl and phenolic‐OH group contents. Whole lignite, humate AG and soil samples, and the HA and FA fractions were analyzed by solid state CP MAS ¹³C NMR and infrared spectroscopy. Carbon, H, and N contents were determined by chemical analysis. NMR spectra of the combined HA+FA extracts resembled the spectra of the whole materials. No additional signals were detected, indicating that alien compounds were not produced during the extraction. The best spectra were obtained with HA samples produced by the NaOH method. These spectra closely resembled those of the untreated materials. Spectral and chemical differences noticed between the HA (or FA) fractions were attributed more to differences in origin than to the extraction procedure. Aliphatic, aromatic and carboxyl groups were the major components of HA from lignite and humate AG. In contrast, HA from the two Haplaquods were characterized by four major components: the aliphatic, polysaccharide, aromatic, and carboxyl groups. Regardless of origin, all the HA fractions contained similar functional groups, as indicated by their close similarities in infrared spectra.     

Dynamic high-resolution 1H and 31P NMR spectroscopy and 1H T2 measurements in postmortem rabbit muscles using slow magic angle spinning

Postmortem changes in rabbit muscle tissue with different glycogen status (normal vs low) were followed continuously from 13 min postmortem until 8 h postmortem and again 20 h postmortem using simultaneous magic angle spinning (1)H and (31)P NMR spectroscopy together with measurement of the transverse relaxation time, T(2), of the muscle water. The (1)H metabolite spectra were measured using the phase-altered spinning sidebands (PASS) technique at a spinning rate of 40 Hz. pH values calculated from the (31)P NMR spectra using the chemical shifts of the C-6 line of histidine in the (1)H spectra and the chemical shifts of inorganic phosphate in the (31)P spectra confirmed the different muscle glycogen status in the tissues. High-resolution (1)H spectra obtained from the PASS technique revealed the presence of a new resonance line at approximately 6.8 ppm during the postmortem period, which were absent in muscles with low muscle glycogen content. This new resonance line may originate from the aminoprotons in creatine, and its appearance may be a result of a pH effect on the exchange rate between the amino and the water protons and thereby the NMR visibility. Alternatively, the new resonance line may originate from the aromatic protons in tyrosine, and its appearance may be a result of a pH-induced protein unfolding exposing hydrophobic amino acid residues to the aqueous environment. Further studies are needed to evaluate these hypotheses. Finally, distributed analysis of the water T(2) relaxation data revealed three relaxation populations and an increase in the population believed to reflect extramyofibrillar water through the postmortem period. This increase was significantly reduced (p < 0.0001) in samples from animals with low muscle glycogen content, indicating that the pH is controlling the extent of postmortem expulsion of water from myofibrillar structures. The significance of the postmortem increase in the amount extramyofibrillar water on the water-holding capacity was verified by centrifugation, which showed a reduced centrifugation loss in muscles with low preslaughter glycogen status (0.9 vs 1.9%, p = 0.07).     

Quantitative solid-state 13C NMR spectroscopy of organic matter fractions in lowland rice soils

Spin counting on solid‐state ¹³C cross‐polarization (CP) nuclear magnetic resonance (NMR) spectra of two humic fractions isolated from tropical lowland soils showed that only 32–81% of potential ¹³C NMR signal was detected. The observability of ¹³C NMR signal (C_obs) was higher in the mobile humic acid (MHA) than in the calcium humate (CaHA) fraction, and increased with increasing intensity of irrigated rice cropping. NMR observability appeared to be related to the nature of the organic carbon, with phenol‐ and methoxyl‐rich samples having the higher values of C_obs. The Bloch decay (BD) technique provided more quantitatively reliable ¹³C NMR spectra, as evidenced by values of C_obs in the range 91–100% for seven of the eight humic fractions studied. The BD spectra contained considerably more aryl and carbonyl signal, and less O–alkyl and alkyl signal, with the greatest differences between CP and BD spectra observed for the samples with low C_obs(CP). The causes of low CP observability were investigated using the spectral editing technique RESTORE ( RE storation of S pectra via T _CH and T O ne R ho (T_1ρH) E diting). Rapid T_1ρH relaxation was found to be primarily responsible for the under‐representation of carbonyl carbon, whereas inefficient cross‐polarization was primarily responsible for the under‐representation of aryl carbon in CP spectra. Proton NMR relaxation rates T_1H and T_1ρH were found to correlate with other NMR properties and also with cropping management. Non‐uniform rates of T_1H relaxation in two of the CaHA fractions enabled the generation of proton spin relaxation editing subspectra.     

Physicochemical changes in the hull of corn grains during their alkaline cooking

The alkaline cooking of corn in a solution of Ca(OH)2 to produce corn-based foods is oriented to make corn proteins available, to incorporate Ca to the cooked grains, and also to remove the corn hull. This process (nixtamalization) is known in Mexico and Guatemala from prehispanic times; however, the effect of the alkaline cooking on the corn hull remains poorly documented. In this work, the physicochemical changes that take place in the corn hull during its cooking in a saturated solution of Ca(OH)2 were studied using infrared, X-ray diffraction, 13C cross-polarization/magic-angle spinning (CP/MAS) NMR, confocal imaging microscopy, differential scanning calorimetry, and thermogravimetry techniques. The main effect of this treatment on the hull is the removal of hemicelluloses and lignin, increasing the hull permeability and, as a consequence, facilitating the entry of the alkaline solution into the corn kernel. No significant changes were observed in the cellulose fiber network, which remains as native cellulose I, with a crystalline index, according to 13C CP/MAS NMR spectra, of 0.60. The alkaline treatment does not allow the cellulose fibers to swell and their regeneration in the form of cellulose(II). It seems any attempt to make use of the Ca binding capacity of the hull to increase the Ca availability in nixtamalized corn-based foods requires a separated treatment for the hull and kernel. On alkaline cooking, the hull hemicellulose fraction dissolves, losing its ability to bind Ca as a way to incorporate this element into foods elaborated from nixtamalized corn.     

Spin accounting and RESTORE – two new methods to improve quantitation in solid-state 13C NMR analysis of soil organic matter

Rapid T_1ρH relaxation and inefficient cross‐polarization have long been known to affect quantitation in solid‐state ¹³C cross‐polarization (CP) NMR spectra of soil organic matter. We have developed two new techniques to overcome these problems. The first, spin accounting, enables accurate gauging of how quantitative a spectrum is likely to be. The result is expressed as the percentage of potential NMR signal that can be accounted for (C_obs). Spin accounting improves on the established spin counting technique by correcting for rapid T_1ρH relaxation and inefficient cross‐polarization. Spin accounting identifies three components: one that is well represented in CP spectra, one that is under‐represented in CP spectra due to rapid T_1ρH relaxation, and one that is under‐represented in CP spectra due to inefficient cross‐polarization. For a range of eight de‐ashed soils, C_obs was in the range 83–106%, indicating that virtually all potential signal could be accounted for after correcting for rapid T_1ρH relaxation and inefficient cross‐polarization. The second new technique, RESTORE (RE storation of S pectra via T _CH and T O ne R ho (T_1ρH) E diting), generates subspectra for the three components identified in spin accounting. The sum of the three RESTORE subspectra is essentially a corrected CP spectrum. The RESTORE spectra of all eight soils more closely resembled the corresponding, and presumably quantitative, Bloch decay spectra than did the CP spectra. RESTORE identifies the types of structures underestimated by CP, and the cause of their underestimation. Rapid T_1ρH relaxation most affected carbonyl and carbohydrate carbons, whereas inefficient cross‐polarization most affected aromatic carbons.     

Alkyl and other major structures in (13)C-labeled glucose-glycine melanoidins identified by solid-state nuclear magnetic resonance

The high molecular weight fraction of melanoidins formed in the Maillard reaction between isotopically labeled glucose and glycine has been characterized comprehensively using advanced (13)C and (15)N solid-state NMR with spectral editing. We have focused on the fate of glucose in a 1:1 molar ratio with glycine, heated as a coprecipitated powder at 125 °C for 2 h. Quantitative (13)C NMR spectra show that aromatic and alkene carbons make up only 40% of the total in the melanoidin. Spectra of melanoidins made from specifically labeled ((13)C1, (13)C2, (13)C3, and (13)C6) glucose are strikingly different, proving that specific structures of various types are formed. More than half of the glucose-C1 carbons form new C-C bonds, not just C-O and C-N bonds. Most C2 carbons are bonded to N or O and not protonated, while C3 shows the reverse trends. C4 and C5 remain significantly in alkyl OCH sites or become part of heterocyclic aromatic rings. C6 undergoes the least transformation, remaining half in OCH(2) groups. Functional groups characteristic of fragmentation are relatively insignificant, except for N/O-C2 ═ O groups indicating some C(1) + C(5) and C(2) + C(4) fragmentation. On the basis of (13)C-(13)C and (15)N-(13)C correlation spectra, 11 "monomer units" have been identified, including several types of alkyl chain or ring segments, furans, pyrroles, imidazoles, and oxazoles; these are mixed on the nanometer scale. This complexity explains why simple models cannot represent the structure of melanoidins. While none of the "monomer units" represents more than 15% of all C, the 11 units identified together account for more than half of all glucose carbon in the melanoidin.     

Determination of the composition of gentamicin sulfates by 1H and 13C nuclear magnetic spectroscopy

The British Pharmacopoeia test controlling the composition of gentamicin sulfate is based on CW 60 MHz magnetic resonance spectroscopy. Application of this method to FT 90 MHz spectra was evaluated. Results clearly show the limitations of this technique and point out the need for more reliable assay methods. Thus a 13C nuclear magnetic resonance (NMR) procedure for quantitative analysis of gentamicin sulfate was developed. Ratios of 4 gentamicin components (C1, C2, C1a, and C2a) were obtained from peak height measurements of selected resonance signals in spectra recorded under steady-state conditions. Relative response factors were determined from spectra of a reference mixture or, alternatively, from spectra of the individual pure components. Results obtained by the 13C NMR method were in agreement with those obtained by liquid chromatography using pre-column derivatization.     

Biomass characterization of morphological portions of alamo switchgrass

Comparative studies between the leaf and internode portions of switchgrass, Panicum virgatum L., were performed by compositional analysis and structural determination. GC-MS, ICP, and HPAEC-PAD were employed to analyze the chemical compositions of the fractionated switchgrass samples. Quantitative (13)C NMR and CP/MAS (13)C NMR techniques were employed to determine the structures of lignin and cellulose, respectively. These results indicated that the leaves and internodes differed chemically in the amounts of inorganic elements, hot-water extractives, benzene/ethanol extractives, carbohydrates, and lignin content. However, the ultrastructure of isolated cellulose was comparable between leaves and internodes. Ball-milled lignins isolated from leaves and internodes were found to have H/G/S ratios of 12.4/53.9/33.7 and 8.6/54.8/36.6, respectively.     

Solubilization of acyl heterogeneous triacylglycerol in phosphatidylcholine vesicles

The amount of acyl heterogeneous triacylglycerol (TG(HET)) solubilized by phosphatidylcholine (PC) vesicles, prepared by co-sonication of egg PC and small amounts (<6% w/w) of TG(HET), was determined using (13)C nuclear magnetic resonance (NMR). The acyl chains of TG(HET) were predominantly 16 or 18 carbons in length, 50% saturated, and approximately 21.7% (13)C isotopically enriched at the carbonyl carbon. The (13)C NMR spectra revealed two carbonyl resonances at chemical shift values between PC carbonyls and oil-phase TG carbonyls, confirming the presence of TG(HET) solubilized in PC vesicles. Oil-phase TG carbonyl peaks were present only in spectra of vesicles containing >3 wt % TG(HET). Integration of TG(HET) carbonyl resonances determined that PC vesicles solubilized 3.8 wt % of TG(HET), compared to 2.8 wt % of acyl homogeneous triolein. The difference between the maximum solubility of TG(HET) and that of homogeneous TG (TG(HOM)) with similar acyl chain lengths provides evidence that specific acyl composition, in addition to the acyl chain length of triacylglycerols, affects the solubility of TG in PC vesicles and TG-rich lipoprotein surfaces. Thus, TG(HET) may innately be a better model substrate than TG(HOM) for determination of substrate availability of TG at lipoprotein surfaces.     

Studies on the starch and hemicelluloses fractionated by graded ethanol precipitation from bamboo Phyllostachys bambusoides f. shouzhu Yi

Starch from bamboo Phyllostachys bambusoides f. shouzhu Yi evaluated by means of solid-state 13C CP/MAS NMR and X-ray diffraction showed a typical B-type pattern with a very low degree of crystallinity (10.9%). In addition to starch, alkali-soluble hemicelluloses were further fractionated by graded precipitation at ethanol concentrations of 0 (HA), 15, 30, 45, 60, and 75% (v/v). Chemical composition and structural features of the six hemicellulosic subfractions were investigated by a combination of sugar analysis, GPC, FT-IR, GC-MS, 1D (1H and 13C) and 2D (HSQC) NMR spectra, and thermal analysis. The results showed that the bamboo hemicelluloses were O-acetylated 4-O-methyl-glucuronoarabinoxylans (GAX) consisting of a linear (1→4)-β-D-xylopyranosyl backbone decorated with branches at O-3 of α-L-arabinofuranosyl (5-12 mol%) or at O-2 of 4-O-methylglucuronic acid units and acetyl groups (0.8-11 mol%). The molecular weights of these polysaccharides ranged between 13400 and 67500 g/mol, and the molar ratios of A/X and G/X increased with ascending ethanol concentrations. Moreover, xylo-oligosaccharides (XOS) with DP 1-6 were produced by enzymatic hydrolysis of hemicelluloses and the total yields of XOS were range of 21.5 to 40.6%. The structure-property relationships were also established in order to improve enzyme accessibility.     

Organic matter composition in sewage farm soils: Investigations by 13C-NMR and pyrolysis-field ionization mass spectrometry

The composition of soil organic matter (SOM) on sewage farms south of Berlin was investigated by solid-state CP/MAS ¹³C-NMR and pyrolysis-field ionization mass spectrometry (Py-FIMS) of freeze-dried sewage solids and soil samples of differing contamination. These were an untreated soil (USOIL), a former sewage farm used as arable land since 1990 (SF90A), and a recent sewage farm (SF1994). The CP/MAS ¹³C-NMR spectra showed enrichments of the sewage-treated soils with aliphatic C and C in OCH₃-groups and amino acids. In the Py-FI mass spectra the major markers of sewage and SOM in sewage farm soils were (i) N-containing compounds, in particular peptides, (ii) dimethylphthalate (m/z 194), (iii) sterols, and (iv) signals in the mass range m/z 502 to 554 of mono- and diaryl esters which were substituted by long aliphatic chains. The latter signals were intense in the sewage solids, increased in intensity from sample SF90A to SF1994; but they were not present in the USOIL, thus clearly indicating anthropogenic origin. Temperature-resolved Py-FIMS showed that the SOM compounds in the sewage farm soils were generally incorporated into bonds with widely different stabilities which could be relevant for SOM turnover and environmental effects. This is demonstrated for the trapping of dimethylphthalate in a modelled humic substance.     

Effect of gamma-sterilization and autoclaving on soil organic matter structure as studied by solid state NMR, UV and fluorescence spectroscopy

Sterilized soil is often used, for example in degradation studies, sorption experiments, microbiological tests and plant test systems, to distinguish between microbial processes and abiotic reactions. The most commonly used technique for sterilization is autoclaving of the soil. Another technique is irradiation with high‐level gamma radiation (γ‐radiation). One major drawback of sterilization procedures is the possible alteration of the structure of soil components, for example the organic matter. A change in the chemical structure of the soil organic matter can cause different reactions in the above‐mentioned experiments and hence interfere with the aim of clearly distinguishing between biotic and abiotic processes. Two soils (Gleyic Cambisol and Orthic Luvisol) were sterilized by two γ‐irradiation procedures (4 kGy hour^?1 for 9 hours and 1.3 kGy hour^?1 for 27 hours) and repeated autoclaving at 121°C. Gentle physical aggregate fractionation of the sterilized soils revealed a decrease in the aggregation of the soil, which was reflected in an increase of the clay fraction. Subsequent analysis of the aqueous phase revealed much more dissolved organic matter (DOM) in the γ‐sterilized and autoclaved soils than in the untreated soils. Ultraviolet (UV) and fluorescence spectra of the DOM showed a decrease in the aromaticity and polycondensation of the dissolved organic carbon (DOC). ¹³C cross‐polarization/magic‐angle spinning nuclear magnetic resonance (¹³C‐CP/MAS NMR) spectra of the unfractionated soils and their respective soil fractions before and after sterilization showed that the most important change occurred in the carbohydrate and N‐alkyl region, the main components of microorganisms. In general, the impact of the sterilization method was stronger for autoclaving. The γ‐sterilized soils and fractions displayed both fewer and smaller changes in the soil organic matter.