Characterisation of thermally modified hard- and softwoods by C CPMAS NMR

The changes induced by thermal modification in the chemical structure of spruce [Picea abies (L.) Karst.], birch (Betula pendula), aspen (Populus tremula) and oak (Quercus robur) were studied by ¹³C CPMAS NMR spectroscopy. Spruce, birch and aspen were thermally modified at 195 °C and oak at 160 °C, under steam, according to an industrial-scale heat treatment process. In both hard- and softwood samples, ¹³C CPMAS NMR measurements revealed a degradation of less ordered carbohydrates (i.e. hemicelluloses and amorphous cellulose) in the thermally modified wood, which resulted in an increase in the cellulose crystallinity. Furthermore, thermal modification induced changes in the lignin structure by a cleavage of the β-O-4 linkages. In the softwood lignin, a decrease also occurred in the methoxyl group content leading to a more condensed lignin structure.     

Changes in C NMR chemical shifts of DNA as a tool for monitoring drug interactions

The antibiotic drug, netropsin, was complexed with the DNA oligonucleotide duplex [d(GGTATACC)]₂ to explore the effects of ligand binding on the ¹³C NMR chemical shifts of the DNA base and sugar carbons. The binding mode of netrospin to TA-rich tracts of DNA has been well documented and served as an attractive model system. For the base carbons, four large changes in resonance chemical shifts were observed upon complex formation: −0.64 ppm for carbon 4 of either Ado4 or Ado6, 1.36 ppm for carbon 2 of Thd5, 1.33 ppm for carbon 5 of Thd5 and 0.94 for carbon 6 of Thd5. AdoC4 is covalently bonded to a heteroatom that is hydrogen bonded to netropsin; this relatively large deshielding is consistent with the known hydrogen bond formed at AdoN3. The three large shielding increases are consistent with hydrogen bonds to water in the minor groove being disrupted upon netropsin binding. For the DNA sugar resonances, large changes in chemical shifts were observed upon netropsin complexation. The 2′, 3′ and 5′ ¹³C resonances of Thd3 and Thd5 were shielded whereas those of Ado4 and Ado6 were deshielded; the ¹³C resonances of 1′ and 4′ could not be assigned. These changes are consistent with alteration of the dynamic pseudorotational states occupied by the DNA sugars. A significant alteration in the pseudorotational states of Ado4 or Ado6 must occur as suggested by the large change in chemical shift of −1.65 ppm of the C3′ carbon. In conclusion, ¹³C NMR may serve as a practical tool for analyzing structural changes in DNA-ligand complexes.     

NMR studies of [U-C]cyclosporin A bound to human cyclophilin B

NMR data (¹H and ¹³C chemical shifts, NOEs) on [[U-¹³C]cyclosporin A bound to cyclophilin B were compared to previously published data on the [U-¹³C]CsA/CyPA complex [Fesik et al., (1991) Biochemistry 30, 6574–6583]. Despite only 64% sequence identity between CyPA and CyPB, the conformation and active site environment of CsA when bound to CyPA and CyPB are nearly identical as judged by the similarity of the NMR data.     

Sn and C NMR study of some trivinyltin(IV) compounds and their complexes

The¹¹⁹Sn and ¹³C NMR spectra of ten trivinyltin(IV) compounds in solutions of non-coordinating (deuteriochloroform, trideuterionitromethane) and coordinating (hexadeuteriodimethyl sulphoxide) solvents have been studied. From δ(¹¹⁹Sn) chemical shifts and ¹J(¹¹⁹Sn,¹³C) coupling constants an evaluation of the coordination number of the central tin atom and the shape of coordination polyhedra around the tin atom has been carried out. Various effects on the δ(¹³C) chemical shifts of both carbon atoms of the vinyl group are also discussed.     

Isolation and characterisation of cellulose obtained by a two-stage treatment with organosolv and cyanamide activated hydrogen peroxide from wheat straw

Seven wheat straw cellulose preparations were isolated by a two-stage acidic organosolv treatment followed by cyanamide activated hydrogen peroxide bleaching. The effects of concentration of acetic and formic acids on the yield of cellulose and degradation of lignin and non-cellulose polysaccharides were investigated. Organic acids were more effective than alcohols on the degradation of lignin and hemicelluloses. Formic acid/acetic acid/water (30/60/10, v/v/v) system was found to be the most effective in delignification and removal of non-cellulose polysaccharides from the straw and did not have any undesirable effects on cellulose properties such as its intrinsic viscosity. In this case, the treatment removed 94.1% of the original lignin and 76.5% of the original hemicelluloses using 0.1% HCl as a catalyst at 85 °C for 4 h. Cyanamide activated hydrogen peroxide bleaching degraded substantial amounts of residual hemicelluloses and lignin, produced the cellulose samples having a relatively high purity. Under a best condition, a cellulose relatively free of lignin (0.7%) and with intrinsic viscosity of 393 ml g⁻¹ and favourable molar mass (213,940 g mol⁻¹) was obtained. Both unbleached and bleached cellulose preparations were further characterised by FT-IR and CP/MAS ¹³C NMR spectroscopy, and thermal stability.     

Mobility of lipid in complexes of amylose–fatty acids by deuterium and C solid state NMR

Palmitic and lauric acid complexes with amylose were studied by solid state methods: ¹³C CP/MAS NMR, deuterium NMR, X-ray powder diffraction and differential scanning calorimetry (DSC). The crystalline amylose complexes were found to be in a V-type sixfold single chain helix. The melting points of the complexes were over 100°C, at least 40–50°C higher than the melting points of the free fatty acids. CP/MAS ¹³C NMR spectra revealed two resonance peaks at 33.6 and 32.4 ppm for the palmitic acid, which were assigned as free and complexed fatty acid, respectively. A single resonance peak at 32.4 ppm was found for the lauric acid and assigned to the complex. The chemical shift of 32.4 ppm for the complexed fatty acids suggests a combined trans and gauche conformation for the fatty acid chain in the complex. T₁ relaxation measurements on the two palmitic acid resonances show different behavior: a very slow relaxation for the 33.6 ppm and a much faster relaxation (1.2 s) for the 32.4 ppm resonances. The latter was similar to the relaxation of the single resonance of the lauric acid (1.1 s). Temperature dependent deuterium spectra of the amylose–lauric acid and amylose–palmitic acid complexes suggest a complete complexation for the amylose–lauric acid, whereas the amylose–palmitic acid complex is partially disassociated by the thermal treatment. Based on the overall data, a partially disordered model is proposed: an imperfect helix with the fatty acid partly inside and partly out, depending on crystallization conditions and the necessity of placing the carboxyl head outside the V-helix.     

模拟增温对长白山北坡垂直样带森林土壤有机碳稳定碳同位素的影响

分析了长白山北坡垂直样带3种典型原始森林地表凋落物及不同粒径土壤组分中有机质的δ¹³C值,并将在岳桦林样地(EB,海拔1996 m)采集的20 cm土柱分别置换到云冷杉林(SF,海拔1350 m)和阔叶红松林(PB,海拔740 m),云冷杉林样地采集的土柱置换到阔叶红松林中,进行为期1年的野外模拟增温试验.结果表明：3种林型土壤的δ¹³C值均显著高于凋落物的δ¹³C值,凋落物和土壤有机质中的δ¹³C值由地表凋落物向土壤下层逐渐增加,而土壤粒径中有机质的δ¹³C值随粒径减小而增大.3种林型中,凋落物δ¹³C值变化趋势为云冷杉林(-28.3‰)>阔叶红松林(-29.0‰)>岳桦林(-29.6‰),而土壤有机质的δ¹³C值变化趋势为岳桦林(-25.5‰)>阔叶红松林(-25.8‰)>云冷杉林(-26.2‰).在土壤温度增加0.7 ℃～2.9 ℃条件下,土壤及其各粒级的δ¹³C值均呈下降趋势,而且<2 μm粘粒和2～63 μm粉粒δ¹³C值的降幅(0.48‰和0.47‰)高于>63 μm砂粒δ¹³C值的降幅(0.33‰).未来气候变暖可能对储藏在细小颗粒中年龄较长的有机碳带来较大的影响.     

外源输入碳在生物结皮土壤各碳组分中的分配特征

研究外源新输入碳进入生物结皮后在各碳组分间的分配特征,可以为理解生物结皮参与碳地球化学循环过程提供数据支持和理论依据。本研究针对黄土高原典型苔藓生物结皮,借助¹³C脉冲标记技术,精确示踪外源新输入碳在生物结皮碳组分中的分配特征及其与无结皮裸地的差异,揭示生物结皮对碳循环的影响。结果表明: 1)由于生物结皮养分循环速率较慢,且与维管束植物相比,其主要生物成分苔藓的生物量有限,导致生物结皮各碳组分的¹³C丰度值均随时间变化表现相对平稳。2)生物结皮的各碳组分¹³C含量均明显高于无结皮裸地,其有机碳、微生物生物量碳、可溶性有机碳中¹³C含量平均分别为0.258、0.078、0.004 mg·kg^-1,分别比裸地高3.1、18.5、2.6倍,且苔藓植株¹³C含量高达1.45 mg·kg^-1。3)生物结皮改变了有机碳各组分的分配特征,其新同化的碳主要分配于活性有机碳库和结皮生物中,表现为¹³C在微生物生物量碳中的分配率(30.6%)高于可溶性有机碳(1.7%),而苔藓植株的¹³C分配率为20.3%。4)生物结皮中微生物生物量¹³C的转移量和库容量分别是裸地的15.7和19.5倍,但其周转率(每月2.94次)略低于裸地(每月3.30次),相应周转期是裸地的1.1倍。综上,生物结皮改变了土壤有机碳组分的分配特征,提升了碳周转速率,在干旱荒漠生态系统碳循环中的作用不容忽视。     

Effect of organosolv and soda pulping processes on the metals content of non-woody pulps

In this work the effect of different pulping processes (ethyleneglycol, diethyleneglycol, ethanolamine and soda) of tow abounded raw materials (empty fruit bunches - EFB and rice straw) on the ash, silicates and metals (Fe, Zn, Cu, Pb, Mn, Ni and Cd) content of the obtained pulps have been studied. Results showed that pulps obtained by diethyleneglycol pulping process presented lower metals content (756 microg/g and 501 microg/g for EFB and rice straw pulp, respectively) than soda pulps (984 microg/g and 889 microg/g). Ethanolamine pulps presented values of holocellulose (74% and 77% for EFB and rice straw pulp, respectively), alpha-cellulose (74% and 69%), kappa number (18.7 and 18.5) and viscosity (612 and 90 6ml/g) similar to those of soda pulp, and lower lignin contents (11% and 12%).     

长期垄作稻田腐殖质稳定碳同位素丰度(δ13C)分布特征

研究了四川盆地丘陵区连续16年垄（宽垄）作稻田土壤稳定碳库腐殖质组分的稳定碳同位素(δ¹³C)分布特征.结果表明： 稻田土壤有机碳含量为宽垄作＞垄作＞水旱轮作.腐殖质碳以胡敏素为主,占土壤碳含量的21%～30%,提取碳以胡敏酸为主,分别占土壤有机碳和腐殖质的17%～21%和38%～65%.土壤有机碳的δ¹³C值介于-27.9‰～-25.6‰,20～40 cm和0～5 cm土壤有机碳δ¹³C值之差约为1.9‰.土壤胡敏酸δ¹³C值比土壤有机碳低1‰～2‰,更接近于油菜和水稻秸秆及根系的δ¹³C值.土壤富里酸δ¹³C值分别较土壤有机碳和胡敏酸高2‰和4‰.耕作层和犁底层胡敏素δ¹³C值分别介于-23.7‰～-24.9‰和-22.6‰～-24.2‰,δ¹³C值的变化反映了耕层中腐殖质的新老混合现象.各有机组分δ¹³C值递减顺序为：胡敏素＞富里酸＞土壤有机碳＞稻草（油菜）残体＞胡敏酸.长期水稻种植有利于增加土壤有机碳含量,同时,耕作方式影响土壤腐殖质δ¹³C在耕作层和犁底层中的分布格局.     

Synthesis and NMR structural analysis of O-succinyl derivative of low-molecular-weight κ-carrageenan

Low-molecular-weight (LMW) κ-carrageenan was achieved through mild hydrochloric acid hydrolysis of κ-carrageenan. The acylation of LMW κ-carrageenan was performed by use of tetrabutylammonium (TBA) salt of the anionic polysaccharide fragments, succinic anhydride, 4-dimethylaminopyridine and tributylamine under homogeneous conditions in N,N-dimethylformamide at 80 °C. Investigation of FT-IR spectrum of the succinylated LMW κ-carrageenan showed that a monoester derivative with succinyl group was formed when LMW κ-carrageenan reacted with succinic anhydride. The ¹H and ¹³C NMR spectroscopy has been used to characterize the fine structure of O-succinyl derivative of the LMW κ-carrageenan. The ¹³C and ¹H NMR chemical shifts of disaccharide unit of O-succinyl LMW κ-carrageenan have been fully assigned using 2D NMR spectroscopic techniques.     

On hexenuronic acid (HexA) removal and mediator coupling to pulp fiber in the laccase/mediator treatment

Flax soda/AQ pulps were treated with different fungal laccase-mediator combinations followed by physical and chemical characterization of the pulps to obtain a thorough understanding of the laccase/mediator effects on hexenuronic acid (HexA) removal and the coupling of mediator onto pulps for fiber functionalization. Large differences were found and the presence of lauryl gallate (LG) during Trametes villosa laccase (TvL) treatment (TvL + LG) resulted in a much larger reduction of pulp-linked HexA than the combination of p-coumaric acid (PCA) and Pycnoporus cinnabarinus laccase (PcL). A major portion of LG became attached to the pulp as revealed by an increase in the kappa number and further confirmed by thioacidolysis and ¹H NMR analysis of solubilized pulp fractions. Additional experiments with other chemical pulps and isolated pulp xylan and lignin revealed that HexA seems to be the sole pulp component attacked by TvL + LG. As a substrate for TvL, the reaction preference order is PCA > HexA > LG.     

Biosynthesis of C-labeled branched polysaccharides by pestalotiopsis from C-labeled glucoses and the mechanism of formation

Biosynthesis of branched glucan by Pestalotiopsis from media containing D-(1-¹³C)glucose, D-(2-¹³C)glucose, D-(4-¹³C)glucose, D-(6-¹³C)glucose or a mixture of D-(1-¹³C)glucose and D-(2-¹³C)glucose was carried out to elucidate biosynthetic mechanism of branched polysaccharides. ¹³C NMR spectra of the labeled polysaccharides were determined and assigned. Analysis of ¹³C NMR spectra of glucitol acetates obtained from hydrolysates of the labeled branched polysaccharides indicated that transfer of labeling from C-1 to C-3 and C-6 carbons, from C-2 to C-1, C-3 and C-5 carbons, and from C-6 to C-1 carbon. From the results the percentages of routes via which the polysaccharide is biosynthesized are estimated. They show that the biosynthesis of the polysaccharide via the Embden-Meyerhof pathway and that from lipids and proteins are more active, and the pentose cycle is less active, than in the biosynthesis of cellulose and curdlan. As for the results, labeling at C-6 carbon in the branched polysaccharide cultured from D-(6-¹³C)glucose was low, compared to that of cellulose and curdlan.     

Coordination of a disubstituted alkene in a chelated d-metal-alkyl-alkene complex. Evidence for equilibrium between complexes with coordinated and free alkenes

Addition of (Cp*₂YH)₂ (4) to 2-methyl-1,4-pentadiene produced the yttrium-alkyl-alkene chelate complex Cp*₂YCH₂CH₂CH₂C(CH₃)=CH₂ (2) in which a disubstituted alkene is complexed to the metal center. Evidence for coordination of the alkene unit of 2 comes from the ¹H and ¹³C NMR chemical shifts of the vinyl units and from observation of nOe effects between Cp* protons and vinyl hydrogens. The disubstituted alkene ligand of 2 is weakly bound, and evidence for an equilibrium with substantial amounts of complex 3 with a free alkene was obtained from variable temperature ¹H NMR spectroscopy.     

Using simple 13C NMR linewidth and relaxation measurements to make detailed chemical shift assignments in triacylglycerols and related compounds

Two simple experiments measuring the ¹³C linewidths ν_1/2 and spin–lattice relaxation times T₁ of each of the signals in the spectrum of trilinolein indicate that the ν_1/2 and T₁ values are consistent with the different degrees of motional freedom expected for the various ¹³C nuclei. However, for each chain, the ν_1/2 and T₁ measurements indicate a small reversal in mobility at C-10 relative to C-9 before motional freedom again steadily increases on each chain starting at C-11. The T₁ experiment allows unambiguous assignments of the C-8 signal and C-14 signal, which differ by only 0.010 ppm. Measurements of ¹³C ν_1/2 and T₁ values on tripalmitin provide secure assignments for the C-5 and C-6 signals, for which conflicting assignments have been reported. The T₁ measurements also show that among the tightly clustered C-8 through C-12 signals, the C-11 signals are the most downfield, while the C-12 signals are the most upfield, again contrary to a previous report. Similar measurements of ¹³C ν_1/2 and T₁ values on other triacylglycerols or related compounds may prove equally useful in making chemical shift assignments and detecting any discontinuities in motional freedom along a chain. The benefits and possible limitations of ultrahigh field NMR for studying triacylglycerols and related compounds are discussed.     

生长在超干旱环境下的3种相思树种表现出异常低的叶片、树枝、树干、根中δ13C含量

生长在超干旱环境下的3种相思树种表现出异常低的叶片、树枝、树干、根中δ¹³C含量 在植物生理生态学中,叶片中碳13(¹³C)含量负值较少(富集),表明叶片处于通过气孔的气体交换减少,比如在干旱胁迫下。此外,与叶片相比,13C在非光合组织中的负值也较少。然而,对从叶片(光合器官)到树枝、树干和根(非光合器官)中的δ ¹³C数值的关系知之甚少,特别是缺少在关联密切的多个树种间或者不同器官间,以及对生长在极端高温和干旱胁迫下的树木中进行测定。本研究测定了3种近缘沙漠相思树种(Acacia tortilis、A. raddiana和A. pachyceras)从叶片到根的¹³C含量。我们在以色列南部成树的叶片组织中测定了δ ¹³C含量。与此同时,在试验果园进行了为期7年的3个水平的灌溉试验。在试验结束时,测定了叶片、树枝、树干和根的生长参数和δ ¹³C含量。研究结果表明,叶片组织中δ ¹³C含量约为−27‰,其同位素贫化程度远超过生长在地球上最干燥和最热环境中的沙漠树种的预期值。在不同的相思树种和不同器官中,所有灌溉水平处理中的δ ¹³C含量并没有富集(−28‰到ca. −27‰),证实了在成熟相思树中的测定结果。在不同器官中,叶片δ ¹³C含量与树枝和根的δ ¹³C含量异常相似,甚至比树干的δ ¹³C含量负值更少。高度贫化的叶片δ ¹³C表明,尽管这些树木生长在极端干燥的生境中,但其气孔气体交换较高。非光合组织中缺乏δ ¹³C富集可能与叶片和异养组织生长的季节耦合有关。     

鬼箭锦鸡儿叶片和土壤碳稳定同位素特征及其影响因素

为探究高海拔地区的植物碳(C)循环过程与其生境的关系,以生长在高山地区的豆科灌木鬼箭锦鸡儿为研究对象,沿着横跨我国东西部山区的样带采集35个样点的鬼箭锦鸡儿叶片和土壤样品,分析了鬼箭锦鸡儿叶片碳稳定同位素组成(δ¹³C)、土壤δ¹³C、叶片和土壤δ¹³C差值(Δδ¹³C)在不同采样点的特征及其与气候因子、叶片和土壤元素的关系。结果表明:鬼箭锦鸡儿叶片δ¹³C的变化范围为-30.9‰～-27.1‰,平均值为-28.4‰,土壤δ¹³C的变化范围为-26.2‰～-23.2‰,平均值为-25.3‰,Δδ¹³C的变化范围为2.0‰～7.7‰,平均值为3.1‰;叶片δ¹³C显著低于土壤δ¹³C,且随着叶片δ¹³C增加,土壤δ¹³C先降低后升高;叶片δ¹³C与生长季均温和叶片C含量呈显著负相关,土壤δ¹³C与相对湿度和最暖月均温呈显著负相关,与土壤碳∶氮(C∶N)呈显著正相关,随土壤C含量的增加土壤δ¹³C先降低后升高,Δδ¹³C与叶片C含量、土壤C含量和土壤C∶N呈显著正相关;气候因子对叶片δ¹³C和Δδ¹³C具有直接影响,同时也通过对叶片和土壤元素的影响,间接导致叶片δ¹³C、土壤δ¹³C和Δδ¹³C的改变。高海拔地区的气候因子、叶片和土壤元素共同影响鬼箭锦鸡儿的C循环过程。     

Preparation and characterisation of methylcellulose from annual cardoon and juvenile eucalyptus

We have studied the preparation of methylcellulose from annual cardoon and juvenile eucalyptus. The high cellulose and low lignin contents of these plants make them potential alternative resources for cellulose derivation. Their high quality pulps were obtained by the Impregnation Rapid Steam Pulping (IRSP) process and Total Chloride Free (TCF) bleaching sequences using hydrogen peroxide. The bleached pulp was methylated twice in an isopropanol slurry at 60 °C for 22 h after mercerisation in 40% NaOH solution. Yields of water-soluble and alkali-soluble methylcellulose were determined by solvent extraction. Substitution patterns of methylcellulose were determined by ¹³C Nuclear Magnetic Resonance (NMR) spectroscopy. The intrinsic viscosities and solution properties of methylcellulose were measured in either 4% NaOH solution or dimethyl sulphoxide (DMSO). This study shows that annual cardoon and juvenile eucalyptus can produce high-quality methylcellulose, which can be used as alternative resources for the production of methylcellulose.     

Nuclear magnetic resonance spectroscopy of 3 beta,7 beta-dihydroxy-5-cholen-24-oic acid multi-conjugates: unusual bile acid metabolites in human urine

Complete ¹H and ¹³C resonance assignments were made for a new type of 3β,7β-dihydroxy-5-cholen-24-oic acid doubly conjugated with sulfuric acid at C-3 and N-acetylglucosamine at C-7 and its glycine- and taurine-amidated triple-conjugates by the combined use of several homonuclear and heteronuclear shift-correlated 2D NMR techniques. The effects of sulfation at C-3, N-acetylglucosaminidation at C-7, and aminoacyl amidation at C-24 on the ¹H and ¹³C chemical shifts and signal multiplicity were clarified. The shielding data serving to characterize each of the bile acid multi-conjugates are also discussed.     

四川裂腹鱼耳石碳、氧稳定同位素特征

为探讨耳石碳(δ¹³C)、氧(δ¹⁸O)稳定同位素在淡水鱼类群体识别中的作用,本研究以养殖条件下不同年龄组四川裂腹鱼为对象,采用稳定同位素质谱仪进行碳、氧同位素测定,揭示耳石中碳、氧稳定同位素特征,探讨其与环境间的关系. 结果表明:1⁺龄四川裂腹鱼δ¹³C和δ¹⁸O值均与耳石质量无显著相关关系,但在微耳石和星耳石之间存在显著差异;不同年龄四川裂腹鱼微耳石δ¹³C和δ¹⁸O平均值分别为(-9.58±0.06)‰、(-8.33±0.17)‰,其在雌雄个体之间均无显著性差异,但在不同年龄组间存在显著差异. 耳石δ¹⁸O和δ¹³C的关联分析能有效区分四川裂腹鱼不同养殖年龄群体,可作为一种识别淡水鱼类养殖群体的手段.