Phenolic syringyl end groups in 13C-enriched hardwoods detected and quantified by solid-state NMR.

While syringyl units are the most abundant monolignols in hardwood lignin, their phenolic (i.e. hydroxyl) end group concentration has not been measured. In two uniformly 13C-enriched young hardwoods, from beech and oak, the syringyl units were quantitatively investigated by advanced solid-state 13C NMR. Small signals of OH-terminated syringyl units were resolved in spectrally edited two-dimensional 13C-13C NMR spectra of the two hardwoods. Their distinct peak positions predicted based on literature data were validated via the abundant OH-terminated syringyl units in hydrolyzed 13C-beechwood. In a two-dimensional 13C-13C exchange spectrum with diagonal-ridge suppression, a well-resolved peak of phenolic syringyl units was observed at the characteristic C-H peak position of syringyl rings, without significant overlap from guaiacyl peaks. Accurate 13C chemical shifts of regular and end-group syringyl units were obtained. Through spectrally edited 2D NMR after 1H inversion recovery, phenols of condensed tannin complexed with arginine were carefully analyzed and shown to overlap minimally with signals from phenolic syringyl units. The local structure and resulting spin dynamics of ether (chain) and hydroxyl (end-group) syringyl units are nearly the same, enabling quantification by peak integration or deconvolution, which shows that phenolic syringyl end groups account for 2 ± 1 % of syringyl units in beechwood and 5 ± 2 % in oakwood. The observed low end-group concentration needs to be taken into account in realistic molecular models of hardwood lignin structure.

Corrected solid-state 13 C nuclear magnetic resonance peak assignment and side-group quantification of hydroxypropyl methylcellulose acetyl succinate pharmaceutical excipients.

Hydroxypropyl methylcellulose acetyl succinate (HPMCAS) is widely used as a pharmaceutical excipient, making a detailed understanding of its tunable structure important for formulation design. Several recently reported peak assignments in the solid-state 13 C NMR spectrum of HPMCAS have been corrected here using peak integrals in quantitative spectra, spectral editing, empirical chemical-shift predictions based on solution NMR, and full spectrum simulation analogous to deconvolution. Unlike in cellulose, the strong peak at 84 ppm must be assigned to C2 and C3 methyl ethers, instead of regular C4 of cellulose. The proposed assignment of signals at <65 ppm to OCH sites, including C5 of cellulose, could not be confirmed. CH2 spectral editing showed two resolved OCH2 bands, a more intense one from O-CH2 ethers of C6 at >69 ppm and a smaller one from its esters and possibly residual CH2 -OH groups, near 63 ppm. The strong intensities of resolved signals of acetyl, succinoyl, and oxypropyl substituents indicated the substitution of >85% of the OH groups in HPMCAS. The side-group concentrations in three different grades of HPMCAS were quantified.

Soil C dynamics after deforestation and subsequent conversion of arable cropland to grassland in humid temperate areas.

Land use and plant-soil management influence soil organic C stocks and soil properties. This study aimed to identify the main mechanisms by which these factors alter soil organic matter (SOM) dynamics and stocks. Changes in the organic C pools and biochemical quality in different OM compartments were assessed: a) after deforestation and intensive cultivation (SOM loss) and then, b) after the conversion of cropland to grassland (SOM replenishment) in a chronosequence of recovery (1-45 years). Topsoil samples were subjected to physical fractionation to assess the distribution of free particulate OM (POM) and mineral associated OM (MAOM). SOM quality was characterized by 13C NMR spectroscopy, thermal analysis (DSC/TG), and microbial activity was monitored by isothermal microcalorimetry. Deforestation and intensive cultivation led to the loss of 80 % of the C stored in the upper mineral soil (up to 30-35 cm). The POM was almost depleted, MAOM underwent significant losses (>40 %) and all OM compounds, including the aromatic C, were affected. The large and unexpected loss of MAOM can be attributed to the low specific surface soil area and also to the labile (biodegradable) nature of the OM in this fraction. After 45 years, conversion of cropland to grassland recovered 68 % of the C lost in the mineral soil (mainly as MAOM), at an annual rate of 1.25 Mg C ha-1. The present findings showed that the persistence of long-term OM depends on how strongly organic compounds are adsorbed onto mineral surfaces (i.e., the specific surface area) and the biochemical nature of OM compounds. Adequate plant-soil management favoured the replenishment of the MAOM under these experimental conditions, and this fraction was an active pool in terms of C storage and biochemical quality. This study served to test current theories about changes in soil C fractions due to land use changes and soil-plant management.

Crystal structure and characterization of the sulfamethazine-piperidine salt.

Sulfamethazine [N1-(4,6-dimethylpyrimidin-2-yl)sulfanilamide] is an antimicrobial drug that possesses functional groups capable of acting as hydrogen-bond donors and acceptors, which make it a suitable supramolecular building block for the formation of cocrystals and salts. We report here the crystal structure and solid-state characterization of the 1:1 salt piperidinium sulfamethazinate (PPD+·SUL-, C5H12N+·C12H13N4O2S-) (I). The salt was obtained by the solvent-assisted grinding method and was characterized by IR spectroscopy, powder X-ray diffraction, solid-state 13C NMR spectroscopy and thermal analysis [differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA)]. Salt I crystallized in the monoclinic space group P21/n and showed a 1:1 stoichiometry revealing proton transfer from SUL to PPD to form salt I. The PPD+ and SUL- ions are connected by N-H+...O and N-H+...N interactions. The self-assembly of SUL- anions displays the amine-sulfa C(8) motif. The supramolecular architecture of salt I revealed the formation of interconnected supramolecular sheets.

Linking the chemical nature of soil organic carbon and biological binding agent in aggregates to soil aggregate stability following biochar amendment in a rice paddy.

Changes in soil aggregation with biochar amendment have been investigated extensively, but how biochar affects the chemical composition of organic carbon (C) and biological binding agents in aggregates and their linkage with soil aggregate stability remains unclear. Soil samples were collected in a rice paddy treated with 0 (C0, control), 10 t ha-1 (C10), 20 t ha-1 (C20) and 40 t ha-1 (C40) biochar for twenty months. The amount and chemical composition of soil organic C (SOC), microbial abundances and glomalin-related soil protein (GRSP) were determined in bulk soil and four fractions: large macroaggregates (>2000 µm), small macroaggregates (250-2000 µm), microaggregates (53-250 µm), and silt + clay (<53 µm). Our results showed that the proportion of >250 µm water-stable aggregates and mean weight diameter were gradually increased with increasing biochar addition rate. The concentrations of SOC, readily oxidizable C and microbial biomass C increased most in the small macroaggregates, followed by microaggregates under biochar amendment. Increasing biochar addition rate gradually decreased the relative contents of alkyl C, O-alkyl C and carbonyl C, but increased those of aromatic C across the aggregates, resulting in a higher aromaticity and hydrophobicity of SOC with respect to the control. The abundances of bacteria, fungi and archaea and the content of GRSP were significantly enhanced in the large and small macroaggregates under the C40 treatment. The proportion of >250 µm aggregates was significantly correlated with the contents of soil organic C fractions, GRSP and microbial abundance. Structural equation modeling further revealed that changes in SOC hydrophobicity and GRSP content under biochar amendment had significant and direct effects on the soil aggregate size distribution. In summary, our findings suggest that biochar amendment in rice paddy could improve soil aggregation through altering the chemical composition of soil organic C and the abundance of biological binding agents.

Grass waste utilization to alter aggregate-related carbon chemical composition and fungal community structure in apple orchard.

The grass-waste management model affects soil organic carbon (SOC) and the microorganism community structure; however, studies on the relationship between the fungal community structure and the SOC chemical component at the aggregate level are poor. Solid-state 13C NMR and 18 S rDNA methods were used to evaluate the relationship between the SOC chemical composition and fungal community abundance at the aggregate level. Grass mulching significantly increased the percentage of labile carbon O-alkyl C (5.19%-11.79%) and decreased the instability of SOC (1.38-0.69). Microaggregates contained higher alkyl C (33.77%) and lower aromatic C (18.31%), and the A/O-A ratio (1.03) was higher than that of macroaggregates (0.89-0.96). Ascomycota, Basidiomycota and Mortierellomycota dominated the fungal community at the phylum level, and their abundance increased after grass mulching. Microaggregates supported more microbial diversity and richness and were rich in the Ascomycota (36.69%-67.49%) phylum, while LM aggregates were rich in Basidiomycota (5.62%-39.84%). We proved that changes in the O-alkyl C, carbonyl C, aromatic C and alkyl C of SOC chemical components were closely connected to fungal community composition, which together explained the change in fungal composition by 63.81%-71.99% among aggregates. We concluded that alterations in the chemical form of organic carbon were closely related to a change in the soil fungal community. This connection has a positive impact on soil nutrient utilization and SOC conversion in fruit-grass composite ecosystems and promotes the understanding of the relationship between the soil microbial community and nutrient cycling during long-term grass waste utilization.

Polycyclic Aromatics Observed in Enzymatic Lignin by Spectral Characterization and Ruthenium Ion-Catalyzed Oxidation.

It is generally considered that lignin is a three-dimensional amorphous polymer consisting of methoxylated phenylpropane structures. However, high yields of monomer structural units of lignin cannot be obtained through various ways, which inspired us to gain insights into the structures of lignin. Herein, enzymatic lignin (EL) was directly characterized by a solid-state 13C nuclear magnetic resonance spectrometer and Fourier transform infrared spectrometer and then subjected to ruthenium ion-catalyzed oxidation. According to the spectral characterization, it can be inferred that multi-ring aromatic clusters exist in EL because of the aromatic bridgehead carbon ratio of 0.136. Based on the results of ruthenium ion-catalyzed oxidation of the EL, it can be deduced that (1) double- and triple-aromatic ring clusters exist in the EL besides the traditional phenylpropane single-aromatic ring clusters, and (2) some aromatic rings with long-alkyl chain substituents exist in the EL, which is quite different from the traditional cognition of lignin. This investigation provides a new insight into the structure of EL.

Physicochemical Changes in Biomass Chars by Thermal Oxidation or Ambient Weathering and Their Impacts on Sorption of a Hydrophobic and a Cationic Compound.

This study examined conditions that mimic oxidative processes of biomass chars during formation and weathering in the environment. A maple char prepared at the single heat treatment temperature of 500 °C for 2 h was exposed to different thermal oxidation conditions or accelerated oxidative aging conditions prior to sorption of naphthalene or the dication paraquat. Strong chemical oxidation (SCO) was included for comparison. Thermal oxidation caused micropore reaming, with ambient oxidation and SCO much less so. All oxidative treatments incorporated O, acidity, and cation exchange capacity (CEC). Thermal incorporation of O was a function of headspace O2 concentration and reached a maximum at 350 °C due to the opposing process of burn-off. The CEC was linearly correlated with O/C, but the positive intercept together with nuclear magnetic resonance data signifies that, compared to O groups derived by anoxic pyrolysis, O acquired through oxidation by thermal or ambient routes contributes more to the CEC. Thermal oxidation increased the naphthalene sorption coefficient, the characteristic energy of sorption, and the uptake rate due to pore reaming. By contrast, ambient oxidation (and SCO) suppressed naphthalene sorption by creating a more hydrophilic surface. Paraquat sorption capacity was predicted by an equation that includes a CEC2 term due to bidentate interaction with pairs of charges, predominating over monodentate interaction, plus a term for the capacity of naphthalene as a reference representing nonspecific driving forces.

A chemical window into the impact of RNAi silencing of the StNAC103 gene in potato tuber periderms: Soluble metabolites, suberized cell walls, and antibacterial defense.

The growth and survival of terrestrial plants require control of their interactions with the environment, e.g., to defend against desiccation and microbial invasion. For major food crops, the protection conferred by the outer skins (periderm in potato) is essential to cultivation, storage, and marketing of the edible tubers and fruits. Potatoes are particularly vulnerable to bacterial infections due to their high content of water and susceptibility to mechanical wounding. Recently, both specific and conserved gene silencing (StNAC103-RNAi and StNAC103-RNAi-c, respectively) were found to increase the load of wax and aliphatic suberin depolymerization products in tuber periderm, implicating this NAC gene as a repressor of the wax and suberin biosynthetic pathways. However, an important gap in our understanding of StNAC103 silencing concerns the metabolites produced in periderm cells as antimicrobial defense agents and potential building blocks of the deposited suberin biopolymer. In the current work, we have expanded prior studies on StNAC103 silenced lines by conducting comprehensive parallel analyses to profile changes in chemical constituents and antibacterial activity. Compositional analysis of the intact suberized cell walls using solid-state 13C NMR (ssNMR) showed that NAC silencing produced an increase in the long-chain aliphatic groups deposited within the periderm cell walls. LC-MS of polar extracts revealed up-regulation of glycoalkaloids in both StNAC103-RNAi and StNAC103-RNAi-c native periderms but down-regulation of a phenolic amine in StNAC103-RNAi-c and a phenolic acid in StNAC103-RNAi native periderms. The nonpolar soluble metabolites identified using GC-MS included notably abundant long-chain alkane metabolites in both silenced samples. By coordinating the differentially accumulated soluble metabolites and the suberin depolymerization products with the ssNMR-based profiles for the periderm polymers, it was possible to obtain a holistic view of the chemical changes that result from StNAC103 gene silencing. Correspondingly, the chemical composition trends served as a backdrop to interpret trends in the chemical barrier defense function of native tuber periderms, which was found to be more robust for the nonpolar extracts.

Cellulose Structural Changes during Mild Torrefaction of Eucalyptus Wood.

The changes in the cellulose structure of eight Eucalyptus species (E. botryoides, E. globulus, E. grandis, E. maculata, E. propinqua, E. rudis, E. saligna and E. viminalis) in a mild torrefaction (from 160 °C to 230 °C, 3 h) were studied in situ and after cellulose isolation from the wood by solid-state carbon nuclear magnetic resonance (13C NMR), wide angle X-ray scattering (WAXS), Fourier transform infrared spectroscopy (FTIR) and by analytic pyrolysis coupled with gas chromatography and mass spectrometry (Py-GC/MS). Changes in molecular weight were assessed by viscosimetry. A small decrease in cellulose crystallinity (ca. 2%-3%) was attributed to its amorphization on crystallite surfaces as a result of acid hydrolysis and free radical reactions resulting in the homolytic splitting of glycosidic bonds. The degree of the cellulose polymerization (DPv) decreased more than twice during the heat treatment of wood. It has been proposed that changes in the supramolecular structure of cellulose and in molecular weight during a heat treatment can be affected by the amount of lignin present in the wood. The limitations of FTIR and Py-GC/MS techniques to distinguish the minor changes in cellulose crystallinity were discussed.

[Interactions of Fungi Community and Relationship with the Carbon Structure in Arable Mollisols with Gradient Organic Matter Content].

Fungi play an important role in the accumulation and transformation of soil organic matter (SOM) and nutrient cycling. To investigate the relationship between the fungal community and soil organic carbon functional groups under gradient SOM contents in arable mollisols, arable mollisols with 2%-9% SOM content were collected in Northeast China. Solid-state 13C-NMR technology was used to explore the differences in the functional group structure of SOM, and ITS high-throughput sequencing was used to investigate the fungal community structure. The potential interactions between different taxonomic groups of soil fungal community and their associations with organic carbon molecular structures were compared by constructing molecular ecological networks under low SOM (2%-5%) and high SOM (7%-9%) conditions. The 13C-NMR results indicated an increase in the relative abundance of Alkyl C (25.8% to 35.9%). The decrease in Alkyl C/O-Alkyl C indicated a smaller degree of decomposition in high SOM soils. Sordariomycetes and Mortierellomycotina dominated the fungal community and their relative abundance increased with the SOM gradient (P<0.05) from 14.33% to 28.17% and from 7.32% to 23.14%, respectively. The network analysis showed simpler ecological topological properties of the fungal community in low SOM soils, with lower numbers of nodes, edges, and average clustering coefficients than those in high SOM soils. A closer relationship between fungi and organic carbon functional groups, especially LOC, was observed in low SOM soils. The random forest model showed that LOC had the largest amount for fungal interactions in low SOM soils (10%), followed by recalcitrant organic carbon (ROC). In comparison, LOC contributed less to the variations in fungal interactions in high SOM soils (7.4%). With globally increasing soil carbon loss, the limition of the carbon resources, especially the reduction of LOC, may reduce the stability and ecological functions of soil fungal communities.

Using Solid-State 13C NMR Spectroscopy to Study the Molecular Organization of Primary Plant Cell Walls.

A knowledge of the mobilities of the polysaccharides or parts of polysaccharides in a cell-wall preparation provides information about possible molecular interactions among the polysaccharides in the cell wall and the relative locations of polysaccharides within the cell wall. A number of solid-state 13C NMR techniques have been developed that can be used to investigate different types of polysaccharide mobilities: rigid, semirigid, mobile, and highly mobile. In this chapter techniques are described for obtaining spectra from primary cell-wall preparations using CP/MAS, proton-rotating frame, proton spin-spin, spin-echo relaxation spectra and single-pulse excitation. We also describe how proton spin relaxation editing can be used to obtain subspectra for cell-wall polysaccharides of different mobilities, and how 2D and 3D solid-state NMR experiments have recently been applied to plant cell walls.

Effective delignification of lignocellulosic biomass by microwave assisted deep eutectic solvents.

To establish an environmentally friendly and cheaper method to delignify lignocellulosic biomass feedstocks, deep eutectic solvents (DESs) were investigated as a green alternatives to conventional solvents. Six different DESs were facilely prepared and used to delignify miscanthus and birchwood feedstocks, including monocarboxylic acid/choline chloride (ChCl), dicarboxylic acid/ChCl and polyalcohol/ChCl. The enhanced delignification efficiency was evaluated in relation to the nature of the hydrogen bond donors and acid strength of DES. The largest extraction of lignin from the miscanthus and birchwood was achieved using ChCl.formic acid and ChCl.oxalic acid DES, respectively. The TGA and 13C NMR characterization results of the extracted lignin samples indicated that the different types of lignin were produced using different DESs. The reaction optimization results showed an increase in lignin extraction with increasing temperature from 60 to 130 °C. However, the optimal reaction time was different, 30 min for miscanthus and 60 min for birchwood. A convenient and reliable method for the quantification of lignin was developed using UV-Vis spectrophotometry.

A molecular fluorophore in citric acid/ethylenediamine carbon dots identified and quantified by multinuclear solid-state nuclear magnetic resonance.

The composition of fluorescent polymer nanoparticles, commonly referred to as carbon dots, synthesized by microwave-assisted reaction of citric acid and ethylenediamine was investigated by 13 C, 13 C{1 H}, 1 H─13 C, 13 C{14 N}, and 15 N solid-state nuclear magnetic resonance (NMR) experiments. 13 C NMR with spectral editing provided no evidence for significant condensed aromatic or diamondoid carbon phases. 15 N NMR showed that the nanoparticle matrix has been polymerized by amide and some imide formation. Five small, resolved 13 C NMR peaks, including an unusual ═CH signal at 84 ppm (1 H chemical shift of 5.8 ppm) and ═CN2 at 155 ppm, and two distinctive 15 N NMR resonances near 80 and 160 ppm proved the presence of 5-oxo-1,2,3,5-tetrahydroimidazo[1,2-a]pyridine-7-carboxylic acid (IPCA) or its derivatives. This molecular fluorophore with conjugated double bonds, formed by a double cyclization reaction of citric acid and ethylenediamine as first shown by Y. Song, B. Yang, and coworkers in 2015, accounts for the fluorescence of the carbon dots. Cross-peaks in a 1 H─13 C HETCOR spectrum with brief 1 H spin diffusion proved that IPCA is finely dispersed in the polyamide matrix. From quantitative 13 C and 15 N NMR spectra, a high concentration (18 ± 2 wt%) of IPCA in the carbon dots was determined. A pronounced gradient in 13 C chemical-shift perturbations and peak widths, with the broadest lines near the COO group of IPCA, indicated at least partial transformation of the carboxylic acid of IPCA by amide or ester formation.

Understanding the Polymerization of Polyfurfuryl Alcohol: Ring Opening and Diels-Alder Reactions.

Polyfurfuryl alcohol (PFA) is one of the most intriguing polymers because, despite its easy polymerization in acid environment, its molecular structure is definitely not obvious. Many studies have been performed in recent decades, and every time, surprising aspects came out. With the present study, we aim to take advantage of all of the findings of previous investigations and exploit them for the interpretation of the completely cured PFA spectra registered with three of the most powerful techniques for the characterization of solid, insoluble polymers: Solid-State 13C-NMR, Attenuated Total Reflectance (ATR), Fourier Transform Infrared (FTIR) spectroscopy, and UV-resonant Raman spectroscopy at different excitation wavelengths, using both an UV laser source and UV synchrotron radiation. In addition, the foreseen structures were modeled and the corresponding 13C-NMR and FTIR spectra were simulated with first-principles and semi-empiric methods to evaluate their matching with experimental ones. Thanks to this multi-technique approach, based on complementary analytical tools and computational support, it was possible to conclude that, in addition to the major linear unconjugated polymerization, the PFA structure consists of Diels-Alder rearrangements occurring after the opening of some furanic units, while the terminal moieties of the chain involves γ-lactone arrangements. The occurrence of head-head methylene ether bridges and free hydroxyl groups (from unreacted furfuryl alcohol, FA, or terminal chains) could be excluded, while the conjugated systems could be considered rather limited.

Developmental changes in collenchyma cell-wall polysaccharides in celery (Apium graveolens L.) petioles.

BACKGROUND: Collenchyma cells occur widely in eudicotyledons and provide mechanical support for growing organs. At maturity, the cells are elongated and have thick, non-lignified walls, which in celery contain cellulose and pectic polysaccharides, together with xyloglucans and heteroxylans and heteromannans. A previous study suggested that at least some of the collenchyma cell wall in celery is laid down after expansion has stopped and is thus secondary. In the present study, we re-examined this. We used chemical analysis and immunomicroscopy to determine changes in the polysaccharide compositions of these walls during development. Additionally, solid-state NMR spectroscopy was used to examine changes in polysaccharide mobilities during development. RESULTS: We showed the collenchyma walls are deposited only during cell expansion, i.e. they are primary walls. During cell-wall development, analytical and immunomicroscopy studies showed that within the pectic polysaccharides there were no overall changes in the proportions of homogalacturonans, but there was a decrease in their methyl esterification. There was also a decrease in the proportions of the (1 → 5)-α-L-arabinan and (1 → 4)-ß-D-galactan side chains of rhamnogalacturonan I. The proportions of cellulose increased, and to a lesser extent those of xyloglucans and heteroxylans. Immunomicroscopy showed the homogalacturonans occurred throughout the walls and were most abundant in the middle lamellae and middle lamella junctions. Although the (1 → 4)-ß-D-galactans occurred only in the rest of the walls, some of the (1 → 5)-α-L-arabinans also occurred in the middle lamellae and middle lamella junctions. During development, the location of the xyloglucans changed, being confined to the middle lamellae and middle lamella junctions early on, but later occurred throughout the walls. The location of the heteroxylans also changed, occurring mostly in the outer walls in young cells, but were more widely distributed in mature cells. Solid-state NMR spectroscopy showed that particularly cellulose, but also homogalacturonans, decreased in mobility during development. CONCLUSIONS: Our studies showed that celery collenchyma cell walls are primary and that during their development the polysaccharides undergo dynamic changes. Changes in the mobilities of cellulose and homogalacturonans were consistent with the cell walls becoming stiffer as expansion ceases.

Molecular-level elucidation of saccharin-assisted rapid dissolution and high supersaturation level of drug from Eudragit® E solid dispersion.

In this work, the effect of saccharin (SAC) addition on the dissolution and supersaturation level of phenytoin (PHT)/Eudragit® E (EUD-E) solid dispersion (SD) at neutral pH was examined. The PHT/EUD-E SD showed a much slower dissolution of PHT compared to the PHT/EUD-E/SAC SD. EUD-E formed a gel layer after the dispersion of the PHT/EUD-E SD into an aqueous medium, resulting in a slow dissolution of PHT. Pre-dissolving SAC in the aqueous medium significantly improved the dissolution of the PHT/EUD-E SD. Solid-state 13C NMR measurements showed an ionic interaction between the tertiary amino group of EUD-E and the amide group of SAC in the EUD-E gel layer. Consequently, the ionized EUD-E could easily dissolve from the gel layer, promoting PHT dissolution. Solution-state 1H NMR measurements revealed the presence of ionic interactions between SAC and the amino group of EUD-E in the PHT/EUD-E/SAC solution. In contrast, interactions between PHT and the hydrophobic group of EUD-E strongly inhibited the crystallization of the former from its supersaturated solution. The PHT supersaturated solution was formed from the PHT/EUD-E/SAC SD by the fast dissolution of PHT and the strong crystallization inhibition effect of EUD-E after aqueous dissolution.

Synthesis, characterisation and application of pyridine-modified chitosan derivatives for the first non-racemic Cu-catalysed Henry reaction.

The preparation, characterisation and application of two pyridine-modified chitosan derivatives (C1 and C2) containing Cu(OAc)2 adsorbed as catalysts for the conversion of benzaldehyde into 2-nitro-1-phenylethanol are described. Quantitative solid-state 13C multiple-contact cross-polarization, magic-angle-spinning, nuclear magnetic resonance (MC-CP MAS NMR) measurements confirmed the successful grafting of 2-pyridinecarboxaldehyde and 6-methylpyridine-2-carboxaldehyde to the chitosan backbone and indicated that 47(±2)% of the NH2 groups were grafted for both C1 and C2. The use of C1-Cu(OAc)2 as a catalyst in the nitroaldol reaction led to 96(±1)% conversion and 19(±4)% enantiomeric excess (ee), while the use of C2-Cu(OAc)2 as a catalyst also promoted the nitroaldol reaction, affording almost quantitatively the expected 2-nitro-1-phenylethanol (98(±1)%) with 14.5(±1.5)% ee.

Characterization of crystalline linear (1→3)-α-d-glucan synthesized in vitro.

We investigated the crystal structure and molecular arrangement of the linear (1→3)-α-d-glucan synthesized by glucosyltransferase GtfJ cloned from Streptococcus salivarius using sucrose as a substrate. The synthetic products had two morphologies: wavy fibril-like crystals as major and thin lamellae as minor products. Their structures were analyzed using electron microdiffraction, synchrotron X-ray powder diffraction, and solid-state 13C NMR spectroscopy. The fibrils and lamellae had the same allomorphic form but different molecular arrangements. The wet crystals were in a hydrated form, which converted into an anhydrous form with a significant decrease in crystallinity on drying. The hydrated and anhydrous forms had an extended-chain conformation with 2/1 helix, and the hydrated form was estimated to contain one water molecule per glucose residue. The long glucan chains were folded in the fibril crystals, while the short, extended chains were arranged perpendicular to the base plane of the lamellae.

Quantification of gabapentin polymorphs in gabapentin/excipient mixtures using solid state 13C NMR spectroscopy and X-ray powder diffraction.

Gabapentin was used as a model pharmaceutical compound with susceptibility to polymorphic transformation as a function of environmental and mechanical stress. The utility of 13C CP/MAS NMR and XRPD as stability-indicating methods to quantify polymorphic transformation kinetics was investigated. Polymorphic Form II and III were distinguishable based on their chemical shift and distinct diffraction peak differences. Reproducible and accurate quantification of polymorphic composition in the presence of selected excipients was demonstrated using both signals from 13C CP/MAS NMR spectra and XRPD patterns. The effect of excipients on polymorphic transformations (Form II→III) was determined by measuring the transformation after co-milling. Both 13C CP/MAS NMR and XRPD were capable of measuring polymorphic composition in co-milled excipient mixtures without excipient peak interference. The amounts of Form III present in co-milled mixtures containing colloidal silicon dioxide, starch, hydroxy propyl cellulose and dibasic calcium phosphate were 8.7, 21, 33, and 39mol%, respectively. A quenching procedure for obtaining 13C CP/MAS NMR spectra and environmentally-controlled XRPD were devised to determine polymorphic transformation kinetics of co-milled excipient mixtures during storage.