Size Exclusion Chromatography-multiangle Laser Light Research Articles

Effect of sheeting stress and heating on the molecular chain structure, size, and conformation of gluten proteins during noodle processing

In this paper, the changes in molecular structure, size distribution, and conformation of gluten induced by the mechanical stress during sheeting and the heating effect during cooking of noodles were investigated by atomic force microscopy (AFM), X-ray scattering (SAXS), and size-exclusion chromatography multi-angle laser light scattering (SEC-MALLS). AFM results showed that the height and width of gluten molecular chains were decreased after calendering and increased post-cooking. Calendering reduced the molecular size and compacted the gluten network (increased Dm). The SDS extractable gluten components showed a quasi-spherical chain conformation but gradually changed to a random coil conformation due to heating. According to SEC-MALLS profiles, calendering and appropriate heating (4 min) promoted the homogeneity of molecular mass of glutenin polymer. The mechanical depolymerization and the heat-promoted polymerization of gluten could be due to the breakage and formation of disulfide bonds and the interconversion of sulfhydryl groups.

Non-filamentous sludge bulking induced by exopolysaccharide variation in structure and properties during aerobic granulation

The forming mechanism of non-filamentous sludge bulking during aerobic granulation were investigated basing on three feeding strategies (R1 direct aeration after fast feeding, R2 anaerobic stirring after fast feeding and R3 anaerobic plug-flow slow feeding). Results showed that strong selection stress (shortening settling time) led to a sharp flocs washout and the subsequent increase of food to microorganisms (F/M) in R1 and R3 reactors, but not found in R2 due to the different strategies of feeding modes. With the increase of F/M, zeta potential and hydrophobicity of sludge surfaces significantly decreased and thus leading to an enhanced repulsive force and energy barriers for sludge aggregation. Particularly, when F/M exceeded 1.2 kgCOD/(kgMLSS·d), non-filamentous sludge bulking was ultimately triggered in R1 and R3. Further analysis showed that massive extracellular exopolysaccharide (PS) accumulated on the surfaces of non-filamentous bulking sludge due to the increased abundance of the microorganisms related to PS secretion during sludge bulking. In addition, significantly increased intracellular second messenger (c-di-GMP), a key substance regulating PS biosynthesis, was confirmed via its concentration determination as well as microbial function prediction analysis, which played a critical role in sludge bulking. Combing with the systematic detection from surface plasmon resonance system, rheometer and size-exclusion chromatography-multiangle laser light detection-refractive index system, higher molecular weight, compact conformation, higher viscosity and higher hydrophilicity was determined in sludge bulking PS relative to PS extracted from non-filamentous bulking sludge. Clearly, the changes of PS (content, structures and properties) driven by c-di-GMP are the dominant mechanism for the formation of non-filamentous sludge bulking during aerobic granulation. This work could provide theoretical support for successful start-up and application of aerobic granular sludge technology.

Quality Distinguish of Red Ginseng from Different Origins by HPLC–ELSD/PDA Combined with HPSEC–MALLS–RID, Focus on the Sugar-Markers

Red ginseng (RG) has been extensively utilized in Asian countries due to its pharmacological effects. For the quality evaluation of RG, small molecules, such as ginsenosides, have been widely considered as candidates of its quality markers (Q-markers), and various analytical techniques have been developed in order to identify these compounds. However, despite the efforts to analyze the hydrophobic constituents, it is worth pointing out that about 60% of the mass of RG is made of carbohydrates, including mono-, oligo- and polysaccharides. Consequently, the quality differentiation and identification of RG from the perspective of sugar-markers should be focused. High performance liquid chromatography and evaporative light scattering detector (HPLC–ELSD) method for the determination of disaccharides in RG was established. Furthermore, high performance size exclusion chromatography–multi-angle laser light scattering–refractive index detector (HPSEC–MALLS–RID) for the determination of molecular weight and high performance liquid chromatography photodiode array (HPLC–PDA) for the determination of compositional monosaccharides in RG polysaccharides were also established. HPLC–ELSD/PDA combined with HPSEC–MALLS–RID could be used to determine the contents of disaccharides, molecular weights, and compositional monosaccharides of RG polysaccharides, which could be used for quality control, and this is a new view on the sugar marker to quality differentiation of various origins of RG.

Characterisation, Chain Conformation and Antifatigue Effect of Steamed Ginseng Polysaccharides With Different Molecular Weight.

Two polysaccharides were obtained from steamed ginseng via ultrafiltration, and their physical–chemical properties, solution properties and antifatigue activities were studied. WSGP-S3 and WSGP-G3 were acid heteropolysaccharides with the molecular weights of 2.03 × 104 and 4.86 × 104, respectively. They were composed of different molar ratios of the monosaccharides Rha, GlcA, GalA, Glc, Gal, and Ara. The results of size-exclusion chromatography–multiangle laser light scattering analysis, Conge red staining and Circular dichroism spectroscopy revealed that WSGP-S3 exhibited a random conformation of branched clusters in solution. By contrast, WSGP-G3 exhibited an ordered conformation, including helix-like conformations in aqueous solution. Antifatigue activity tests proved that WSGP-S3 markedly prolonged the exhaustive swimming time of fatigued mice; increased liver and muscle glycogen levels and superoxide dismutase, catalase, glutathione peroxidase activities and decreased blood lactic acid, nitrogen and malondialdehyde levels compared with the control treatment. Moreover, it enhanced spleen cell proliferation in fatigued mice. By contrast, WSGP-G3 had no significant effect on fatigued mice. The results showed that WSGP-S3 might have a major contribution to the antifatigue effects of steamed ginseng polysaccharides and could be a potential anti-fatigue polysaccharide.

Short- and Long-Term Effects of X-ray Synchrotron Radiation on Cotton Paper.

X-ray analytical techniques are increasingly being used to study manuscripts and works of art on paper, whether with laboratory equipment or synchrotron sources. However, it is difficult to anticipate the impact of X-ray photons on paper- and cellulose-based artifacts, particularly due to the large variety of their constituents and degradation levels, and the subsequent material multiscale heterogeneity. In this context, this work aims at developing an analytical approach to study the modifications in paper upon synchrotron radiation (SR) X-ray radiation using analytical techniques, which are fully complementary and highly sensitive, yet not frequently used together. At the molecular scale, cellulose chain scissions and hydroxyl free radicals were measured using chromatographic separation techniques (size-exclusion chromatography-multiangle laser light scattering-differential refractive index (SEC-MALS-DRI) and reversed-phase high-performance liquid chromatography-fluorescence detector-diode array detector (RP-HPLC-FLD-DAD)), while the optical properties of paper were characterized using spectroscopy (UV luminescence and diffuse reflectance). These techniques showed different sensitivities toward the detection of changes. The modifications in the cellulosic material were monitored in real time, within a few days, and up to 2 years following the irradiation to define a lowest observed adverse effect dose (LOAED). As paper is a hygroscopic material, the impact of the humidity in the environment was studied using this approach. Three levels of moisture content in the paper, achieved by conditioning the samples and irradiating them at different relative humidities (RHs), were studied (0, 50, 80% RH). It was shown that very low moisture content accelerated molecular and optical modifications.

Synthesis of Novel pH-Tunable Thermoresponsive Hydroxyl-Terminated Hyperbranched Polyether.

In this study, a new pH-tunable thermoresponsive hydroxyl-terminated hyperbranched polyether (HTHP 2) was successfully prepared via a one-pot cationic polymerization technique and postmodification. In the first step, hydroxyl-terminated hyperbranched polyether containing double bonds (HTHP 1) were synthesized. Then, through thiol-ene “click” reaction, pH-responsive carboxyl groups were introduced to the target polymer of HTHP 2. The products were characterized via Fourier-transform infrared spectra (FTIR), nuclear magnetic resonance (NMR), and size-exclusion chromatography-multiangle laser light scattering (SEC-MALLS). Moreover, dynamic light scattering (DLS) and UV–Vis spectroscopy was employed to study the pH- and thermoresponsiveness in detail. Results showed that HTHP 2 possessed typical pH-controllable thermoresponsive behavior. By regulating the solution pH value range 3.0–5.4, LCST of HTHP 2 could be changed from 12.8 to 68.0 °C. Meanwhile, the cell viabilities of A549 cells were more than 80% for in vitro cytotoxicity tests of HTHP 2, suggested that HTHP 2 polymers are of good biocompatibility for up to 24 h.

Thermodynamic Stability of the Transcription Regulator PaaR2 from Escherichia coli O157:H7

PaaR2 is a putative transcription regulator encoded by a three-component parDE-like toxin-antitoxin module from Escherichia coli O157:H7. Although this module’s toxin, antitoxin, and toxin-antitoxin complex have been more thoroughly investigated, little remains known about its transcription regulator PaaR2. Using a wide range of biophysical techniques (circular dichroism spectroscopy, size-exclusion chromatography-multiangle laser light scattering, dynamic light scattering, small-angle x-ray scattering, and native mass spectrometry), we demonstrate that PaaR2 mainly consists of α-helices and displays a concentration-dependent octameric build-up in solution and that this octamer contains a global shape that is significantly nonspherical. Thermal unfolding of PaaR2 is reversible and displays several transitions, suggesting a complex unfolding mechanism. The unfolding data obtained from spectroscopic and calorimetric methods were combined into a unifying thermodynamic model, which suggests a five-state unfolding trajectory. Furthermore, the model allows the calculation of a stability phase diagram, which shows that, under physiological conditions, PaaR2 mainly exists as a dimer that can swiftly oligomerize into an octamer depending on local protein concentrations. These findings, based on a thorough biophysical and thermodynamic analysis of PaaR2, may provide important insights into biological function such as DNA binding and transcriptional regulation.

Therapeutic effects of polysaccharides from Anoectochilus roxburghii on type II collagen-induced arthritis in rats

Anoectochilus roxburghii, a famous Chinese herbal medicine, has been commonly used for the treatment of liver disease, diabetes, and rheumatoid arthritis. Our study aimed to investigate the anti-rheumatoid arthritis effects of A. roxburghii polysaccharides (ARP), using the rat's model of type II collagen-induced arthritis (CIA). ARP was prepared by alcohol sedimentation and structurally characterized based on combined chemical, chromatographic and spectroscopic methods. High Performance Size Exclusion Chromatography-Multiangle Laser Light Scattering-Refrative Index (HPSEC–MALLS–RI) analysis revealed that ARP includes two peaks, and the weight-average molecular weight (Mw) of the principal one was estimated as 5.90 kDa with a relative content of 98.2%. Pharmacological results exhibited that ARP significantly decreased the arthritis index and ameliorated the inflammatory cell infiltration and the synovial tissue destruction in CIA rats. Additionally, ARP possessed significant NO production inhibitory effects and antioxidant activity. Further anti-inflammatory mechanism investigations indicated that ARP significantly inhibited the activation of nuclear factor κB (NF-κB) pathway by suppressing the phosphorylation of IκB and p65, which subsequently down-regulated the mRNA expressions of IL-1β and IL-6 in LPS-stimulated RAW 264.7 cells. These findings suggested that ARP has great potential in the development of functional foods and dietary supplements for the treatment of rheumatoid arthritis.

Molar Mass Analysis of Hydrophobically Modified Hyaluronic Acid by SEC‐MALLS: Facing the Challenges of Amphiphilic Biomacromolecules

AbstractA size exclusion chromatography–multiangle laser light scattering (SEC‐MALLS) method is developed for determining the molar masses of amphiphilic hyaluronan (HA), consisting of a hydrophilic polymer backbone modified to varying extents with hydrophobic, polymerizable acrylate groups. The polymers are prepared via the base‐catalyzed esterification of HA with acrylic anhydride. Various parameters are investigated including mobile phase composition, temperature, pH, salt type, and concentration in order to find the most suitable conditions for completely dissolving the modified polymers irrespective of the degree of substitution (DS), and for suppressing secondary polymer interactions. The hydrolytic and the thermal stability of the different materials at various experimental conditions are also extensively investigated. HAs with DS varying from 0 to 3.4 are subsequently analyzed at the optimized experimental conditions for the mobile phase being DMSO/H2O 60/40(v/v) with 0.05 M LiBr and an operating temperature of 40 °C. The refractive index increments (dn/dc) of the HAs are determined and the results show a dependence of dn/dc on DS.

Characterization of the chemical structures and physical properties of exopolysaccharides produced by various Streptococcus thermophilus strains

Exopolysaccharides (EPS) produced by some lactic acid bacteria are often used by the dairy industry to improve the rheological and physical properties of yogurt, but the relationship between their structure and functional effect is still unclear. The EPS from different species, or different strains from the same species, may differ in terms of molar mass, repeating unit structure, and EPS yield during fermentation of milk. This study aimed to characterize the detailed properties of EPS produced from 7 strains of Streptococcus thermophilus, which is one of the key cultures used for yogurt manufacture. Milk was fermented with strains DGCC 7698, DGCC 7710, DGCC 7785, ST-10255y, St-143, STCth-9204, and ST4239. These strains were selected because they have been used in previous studies on yogurt texture, but a complete description of their EPS structural properties has not yet been reported. All strains were fermented under a similar acidification rate by adjusting the level of supplementation with peptone or the inoculation level, which allowed for a comparison of EPS yields under similar growth conditions (reconstituted skim milk at 40°C). The EPS from each strain was isolated and the weight-average molar mass and z-average root mean square radius determined using size-exclusion chromatography multiangle laser light scattering. The monosaccharide composition of EPS was determined using gas chromatography-mass spectrometry, and repeating unit structure was determined using nuclear magnetic resonance spectroscopy. The weight-average molar mass values of EPS ranged from 0.14 to 1.61 × 106 g/mol. All 7 EPS samples were uncharged. The strains ST-10255y and ST4239 had EPS with the same repeating unit structure. The monosaccharide compositions of the various EPS were mainly composed of glucose and galactose, with low levels of rhamnose in the EPS isolated from DGCC 7698, and N-acetylgalactosamine in the EPS from DGCC 7785, ST-10255y, and ST4239. The yields of EPS (measured when fermented milks reached pH 4.6) ranged from 8.0 to 76.4 mg of glucose equivalents/kg. In addition to (free) EPS, some strains were also able to produce capsular polysaccharide (associated with the bacterial cells) when observed with negative staining technique. The results of our study will help the dairy industry to better understand the mechanism by which different strains of Streptococcus thermophilus affect yogurt texture.

Preparation of Low Molecular Weight Chondroitin Sulfates, Screening of a High Anti-Complement Capacity of Low Molecular Weight Chondroitin Sulfate and Its Biological Activity Studies in Attenuating Osteoarthritis.

Chondroitin sulfate (CS) plays important roles in the complement system. However, the CS structure is complicated due to different sources and the number and positions of sulfate groups. The objective of this study was to prepare different low molecular weight chondroitin sulfates (LMWCSs) and to investigate the biological activity in anti-complement capacity. A series of LMWCSs was prepared from different sources and characterized by ultraviolet-visible (UV) spectroscopy, high-performance liquid chromatography (HPLC), size exclusion chromatography-multiangle laser light scattering (SEC-MALLS) and nuclear magnetic resonance (NMR) spectroscopy. Hemolytic, anti-complement 3 deposition capacity and cell viability assays were carried out to investigate the biological activities in vitro. The results showed that LMWCS prepared from shark cartilage with the oxidative degradation method (LMWCS-S-O) had the best anti-complement capacity. LMWCS-S-O could inhibit the alternative pathway of the complement system and protect chondrocytes from cell death. The attenuating effect of LMWCS-S-O on Osteoarthritis (OA) was investigated by destabilization of the medial meniscus (DMM) model in vivo. Functional wind-up, histological and C5b-9 analyses were used to evaluate the treatment effect on the OA model. In vivo results showed that LMWCS-S-O could attenuate OA. LMWCS-S-O with a high content of ΔDi-2,6diS and ΔDi-6S could be used for attenuating OA through regulating the complement system.

Dietary flavonoid fisetin increases abundance of high-molecular-mass hyaluronan conferring resistance to prostate oncogenesis.

We and others have shown previously that fisetin, a plant flavonoid, has therapeutic potential against many cancer types. Here, we examined the probable mechanism of its action in prostate cancer (PCa) using a global metabolomics approach. HPLC-ESI-MS analysis of tumor xenografts from fisetin-treated animals identified several metabolic targets with hyaluronan (HA) as the most affected. Efficacy of fisetin on HA was then evaluated in vitro and also in vivo in the transgenic TRAMP mouse model of PCa. Size exclusion chromatography-multiangle laser light scattering (SEC-MALS) was performed to analyze the molar mass (Mw) distribution of HA. Fisetin treatment downregulated intracellular and secreted HA levels both in vitro and in vivo Fisetin inhibited HA synthesis and degradation enzymes, which led to cessation of HA synthesis and also repressed the degradation of the available high-molecular-mass (HMM)-HA. SEC-MALS analysis of intact HA fragment size revealed that cells and animals have more abundance of HMM-HA and less of low-molecular-mass (LMM)-HA upon fisetin treatment. Elevated HA levels have been shown to be associated with disease progression in certain cancer types. Biological responses triggered by HA mainly depend on the HA polymer length where HMM-HA represses mitogenic signaling and has anti-inflammatory properties whereas LMM-HA promotes proliferation and inflammation. Similarly, Mw analysis of secreted HA fragment size revealed less HMM-HA is secreted that allowed more HMM-HA to be retained within the cells and tissues. Our findings establish that fisetin is an effective, non-toxic, potent HA synthesis inhibitor, which increases abundance of antiangiogenic HMM-HA and could be used for the management of PCa.

An Intrinsically Disordered Region in the Proapoptotic ASPP2 Protein Binds to the Helicobacter pylori Oncoprotein CagA.

The leading risk factor for gastric cancer in humans is infection by Helicobacter pylori strains that express and translocate the oncoprotein CagA into host epithelial cells. Once inside host cells, CagA interacts with ASPP2, which specifically stimulates p53-mediated apoptosis and reverses its pro-apoptotic function to promote ASPP2-dependent degradation of p53. The X-ray crystal structure of a complex between the N-terminal domain of CagA and a 56-residue fragment of ASPP2, of which 22 residues were resolved, was recently described. Here, we present biochemical and biophysical analyses of the interaction between the additional regions of CagA and ASPP2 potentially involved in this interaction. Using size exclusion chromatography-multiangle laser light scattering, circular dichroism, and nuclear magnetic resonance analyses, we observed that the ASPP2 region spanning residues 331-692, which was not part of the ASPP2 fragment used for crystallization, is intrinsically disordered in its unbound state. By surface plasmon resonance analysis and isothermal titration calorimetry, we found that a portion of this disordered region in ASPP2, residues 448-692, binds to the N-terminal domain of CagA. We also measured the affinity of the complex between the ASPP2 fragment composed of residues 693-918 and inclusive of the fragment used for crystallization and CagA. Additionally, we mapped the binding regions between ASPP2 and CagA using peptide arrays, demonstrating interactions between CagA and numerous peptides distributed throughout the ASPP2 protein sequence. Our results identify previously uncharacterized regions distributed throughout the protein sequence of ASPP2 as determinants of CagA binding, providing mechanistic insight into apoptosis reprogramming by CagA and potential new drug targets for H. pylori-mediated gastric cancer.

Isolation, structural characterization and neurotrophic activity of a polysaccharide from Phellinus ribis

A new polysaccharide named PRG was isolated from the fruiting bodies of Phellinus ribis by hot water extraction, ethanol precipitation, anion-exchange and gel-filtration chromatography. PRG was homogeneous, with a molecular weight of 5.16×103Da, as determined by high-performance size-exclusion chromatography–multiangle laser light scattering analysis. Its structural characteristics were investigated and elucidated by methylation analysis, partial acid hydrolysis, gas–liquid chromatography mass spectrometry, Fourier transform infrared and nuclear magnetic resonance spectroscopy. Based on obtained data, PRG was found to be a β-d-glucan containing a (1→3)-linked backbone, with a branch of two (1→6)-linked and one terminal glucoses substituting at the C-6 position every three residues, along the main chain. PRG exhibited neurotrophic activity, which significantly promoted the neurite outgrowth of the nerve growth factor-stimulated PC12 cells, suggesting that it might be a potential candidate for the treatment of neurodegenerative diseases.

Dimerization of matrix protein is required for budding of respiratory syncytial virus.

Respiratory syncytial virus (RSV) infects epithelial cells of the respiratory tract and is a major cause of bronchiolitis and pneumonia in children and the elderly. The virus assembles and buds through the plasma membrane, forming elongated membrane filaments, but details of how this happens remain obscure. Oligomerization of the matrix protein (M) is a key step in the process of assembly and infectious virus production. In addition, it was suggested to affect the conformation of the fusion protein, the major current target for RSV antivirals, in the mature virus. The structure and assembly of M are thus key parameters in the RSV antiviral development strategy. The structure of RSV M was previously published as a monomer. Other paramyxovirus M proteins have been shown to dimerize, and biochemical data suggest that RSV M also dimerizes. Here, using size exclusion chromatography-multiangle laser light scattering, we show that the protein is dimeric in solution. We also crystallized M in two crystal forms and show that it assembles into equivalent dimers in both lattices. Dimerization interface mutations destabilize the M dimer in vitro. To assess the biological relevance of dimerization, we used confocal imaging to show that dimerization interface mutants of M fail to assemble into viral filaments on the plasma membrane. Additionally, budding and release of virus-like particles are prevented in M mutants that fail to form filaments. Importantly, we show that M is biologically active as a dimer and that the switch from M dimers to higher-order oligomers triggers viral filament assembly and virus production. Human respiratory syncytial virus (RSV) is the most frequent cause of infantile bronchiolitis and pneumonia. The enormous burden of RSV makes it a major unmet target for a vaccine and antiviral drug therapy. Oligomerization of the matrix protein is a key step in the process of assembly and production of infectious virus, but the molecular mechanism of RSV assembly is still poorly understood. Here we show that the RSV matrix protein forms dimers in solution and in crystals; the dimer is essential for formation of higher-order oligomers. Destabilizing the dimer interface resulted in the loss of RSV filament formation and a lack of budding of virus-like particles. Importantly, our findings can potentially lead to new structure-based RSV inhibitors targeting the assembly process.

Characterization of Exopolysaccharides Produced by Bifidobacterium longum NB667 and Its Cholate-Resistant Derivative Strain IPLA B667dCo

Bifidobacteria are natural members of the human intestinal microbiota and some strains are being used as probiotics. Adaptation to bile can allow them to increase survival in gastrointestinal conditions, thus improving their viability. Bifidobacterium longum NB667 and the cholate-resistant strain B. longum IPLA B667dCo produced exopolysaccharides (EPS) that were partially characterized. Analysis by size exclusion chromatography-multiangle laser light scattering indicated that the EPS crude fractions of both strains contained two polymer peaks of different molar mass. On the basis of chromatographic techniques both peaks appeared to be heteropolysaccharides. The smaller peak was mainly composed of glucose, galactose and rhamnose whose molar ratios and linkage types showed slight variations between the EPS fractions of both strains. The bigger peak consisted of glucose and galactose; the monosaccharide composition was identical in the EPS fractions of the two microorganisms, but their infrared spectra presented some differences regarding compounds other than carbohydrates that seem to be associated to the polymer. Differences in the composition of EPS fractions did not affect the capability of crude EPS from B. longum to be fermented by the human intestinal microbiota in fecal batch cultures.

Comb-shaped graft copolymers with cellulose side-chains prepared via click chemistry

Abstract Comb-shaped copolymers with cellobiose acetate or cellulose triacetate (CTA) side-chains, PPMA-g-(CTA2-C15) and PPMA-g-(CTA13-C15), were prepared by grafting N-(15-azidopentadecanoyl)-2,3,6-tri-O-acetyl-4-O-(2,3,4,6-tetra-O-acetyl-β- d -glucopyranosyl)-β- d -glucopyranosylamine (CTA2-C15-N3) and N-(15-azidopentadecanoyl)-tri-O-acetyl-β-cellulosylamine (CTA13-C15-N3, number average degree of polymerization (DPn) = 13) onto poly(2-propyn-1-yl methacrylate) (PPMA, weight average degree of polymerization (DPw, X + Y = 5.59 × 102)) via “click chemistry”. The copolymers were characterized by 1H, 13C and two-dimensional NMR and size exclusion chromatography–multi-angle laser light scattering (SEC–MALS) measurements. The numbers of CTA side-chains (X) of PPMA-g-(CTA2-C15) and PPMA-g-(CTA13-C15) were calculated as 4.03 × 102 and 2.45 × 102, respectively. Copolymers with cellulosic side-chains, PPMA-g-(CELL2-C15) and PPMA-g-(CELL13-C15), were successfully obtained after deacetylation of PPMA-g-(CTA2-C15) and PPMA-g-(CTA13-C15), respectively. X-ray diffraction measurements revealed that PPMA-g-(CELL13-C15) showed crystalline pattern of cellulose II, which is believed to have anti-parallel orientation.

Compared molecular characterization of hyaluronan using multiple-detection techniques

The biological functions as well as physico-chemical properties of hyaluronan (HA), one of the most important and ubiquitous glycosaminoglycan in various organisms, are closely related to its molecular weight, molecular weight distribution and chain conformation. For this reason it is crucially important to make a reliable characterization of these parameters for HA in both chemical and clinical fields. The present work compared the application of different techniques including capillary viscometry, concentration gradient multiangle laser light scattering (CG-MALLS), size exclusion chromatography-multiangle laser light scattering (SEC-MALLS), asymmetric flow field flow fractionation-multiangle laser light scattering (AFlFFF-MALLS) and horizontal agarose gel electrophoresis (AGE), to the characterization of a range of HA samples of bacterial and animal sources. The advantage and limitation of these techniques were discussed in terms of producing accurate and reliable molecular parameters.

Effect of nucleating agents on the molar mass distribution and its correlation with the isothermal crystallization behavior of poly(L‐lactic acid)

AbstractIn this work, the effect of the processing conditions on the thermal behavior of a commercial poly(L‐lactic acid) (PLLA) was investigated. In particular, PLLA having a very high molecular weight was processed by a discontinuous mixer at different mixing conditions. The molar mass variation was investigated by size exclusion chromatography–multiangle laser light scattering chromatography, and the thermal characteristics were detected by the conventional differential scanning calorimetry and dynamic mechanical thermal analysis. Tensile tests were performed at room temperature and at 60°C. Our attention was essentially focused on the crystallization behavior of the various samples obtained. A modest decline in the molar mass was obtained with two mixing temperatures of 210 and 250°C, but a significant increase in the crystalline phase content was noticed, with a concomitant tremendous increment in the crystallization rate. A correlation was also made between the molar mass distribution and crystallization rate in isothermal conditions, where clearly visible were the effects of the processing temperature on both the molar mass distribution and how the nucleating agents affected the crystallinity of PLLA. © 2011 Wiley Periodicals, Inc. J Appl Polym Sci, 2011

Characterization of resulting network polymer precursors in stepwise crosslinking diepoxide/diamine polymerization

Our previous mechanistic discussion of network formation in chainwise crosslinking multiallyl polymerization was extended to stepwise crosslinking diepoxide/diamine polymerization, typically including bisphenol-A diglycidyl ether (BADGE) and 4,4′-diaminodiphenylmethane (DDM). In allyl polymerization a monomer chain transfer is an essential termination reaction, providing only oligomeric primary polymer chains. Therefore, crosslinking multiallyl polymerization could be in the category of a classical gelation theory. Thus, the gelation behavior was discussed by comparing the actual gel point with the theoretical one. Then the resulting network polymer precursors (NPPs) were characterized by size-exclusion chromatography-multiangle laser light scattering-viscometry to clarify the stepwise crosslinking BADGE/DDM polymerization mechanism. Notably, the intrinsic viscosity ratio [η]NPP/[η]Linear tended to decrease with the progress of crosslinking and finally, it reached less than 0.2. This suggests that the structure of resulting NPP becomes dendritic at a conversion close to the gel point. These dendritic NPPs can collide with each other to form crosslinks between NPPs, eventually leading to gelation as a reflection of the high concentration of NPP. The dilution effect on gelation was marked in polar solvent; no gelation was observed at a dilution of 1/5. However, in nonpolar solvent the gelation was promoted by dilution; this is ascribed to enhanced crosslink formation between NPPs through hydrogen bonding due to abundant hydroxyl groups in the NPP generated by the polyaddition reaction. Finally, the subject of “Is cured epoxy resin inhomogeneous?” is briefly discussed. © 2011 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem, 2011