Modulating storage stability of binary gel by adjusting the ratios of starch and kappa-carrageenan.

This study aimed to investigate the interaction mechanism of the waxy starch and kappa carrageenan (KC) gel with different ratios during co-gelatinization and storage. Water distributions, mobilities and rheological properties of the mixture gels were studied. When KC concentration was low (0.5%KC and 4% starch), the starch dominated the system, and the gel strength was the lowest. When KC concentration increased to 0.75%, the gel had the lowest change rate of fracture force, and the most homogeneous network of the freeze-dried gel was observed. When KC concentration increased to 1.0%, the gel strength was high, but the uneven structure led to the instability of the gel. Overall, the gel with 0.75%KC and 4% starch was the most stable during storage, and the exclusion of the two components to each other was the weakest, resulting in the uniform structure of the gel.

Synthesis, characterization and potential application of hydrophobically modified carrageenan derivatives as pharmaceutical excipients.

New amphiphilic derivatives of kappa-carrageenan (KC) were synthesized by hydrophobic modification with octyl chloride. Two different methods based on microwave and conventional heating were used. All KC derivatives (KCRs) were characterized by different techniques. The FT-IR and 1HNMR studies demonstrated that the octyl groups were effectively grafted onto KC backbone and confirmed that the derivative KCRMM, obtained by microwave heating, presented a higher degree of substitution (DS = 0.77) compared to KCRCM (0.45). The amphiphilic character investigation also revealed that KCRMM exhibited a lower critical aggregation concentration (CAC) value of 0.13% w/v than KCRCM (0.15%). Furthermore, KCRs greatly improved the stability of oil/water emulsions as the droplet size decreased with increasing DS. Unlike the conventional heating method, novel derivatives with a higher DS, greater amphiphilic character, and an improved emulsifying power were obtained by microwave-assisted synthesis. These derivatives could potentially be used in food, cosmetics, or as excipients in pharmaceutics.

Physical, structural and thermal properties of composite edible films prepared from pearl millet starch and carrageenan gum: Process optimization using response surface methodology.

The present study aimed at developing edible composite films from pearl millet starch (PMS) and carrageenan gum (CG) blends. Central composite rotatable design of response surface methodology was used to optimize concentrations of PMS (2-5%), carrageenan (1-3.5%) and glycerol (15-35%) for the development of composite film. The optimization was done on the basis of responses viz. thickness, water vapor permeability (WVP), solubility and tensile strength (TS). Both starch and carrageenan concentrations positively affected the TS of films, whereas the reverse trend was observed with increased concentration of glycerol. The optimum level of different parameters resulting in a composite film with maximum TS and lower WVP and solubility values were obtained under conditions of 4.9% starch, 5% carrageenan and 15.73% plasticizer concentrations. The melting temperature of optimized PMS/CG film was higher than PMS film. Morphological studies exhibited homogeneous structure of PMS film while PMS/CG composite film had rough, coarser and slightly irregular surface. The stretched regions in Fourier-transform infrared spectroscopy and Differential Scanning Calorimeter analysis confirmed the miscibility and thermal stability of different components of PMS/CG composite films. The results indicated that the developed optimized PMS/CG film possessed lower WVP, solubility but higher transparency and TS in comparison to PMS film.

Primary, Secondary, Tertiary and Quaternary Structure Levels in Linear Polysaccharides: From Random Coil, to Single Helix to Supramolecular Assembly.

Polysaccharides are ubiquitous in nature and represent an essential class of biopolymers with multiple levels of conformation and structural hierarchy. However, a standardized structural nomenclature, as in the case of proteins, is still lacking due to uncertainty on their hierarchical organization. In this work we use carrageenans as model polysaccharides to demonstrate that several structural levels exist and can be unambiguously resolved by statistical analysis on high resolution Atomic Force Microscopy images, supported by spectroscopic, X-ray scattering and rheological techniques. In direct analogy with proteins, we identify primary, secondary, tertiary and quaternary structures. The structure-property relationship induced by monovalent ions for κ-, ι- and the non-gelling control λ-carrageenan is established from the single chain regime to the occurrence of hydrogels at higher concentrations. For κ-carrageenan in the presence of potassium, a disorder-order transition from random coil to single helix is first observed (secondary structure), followed by intrachain supercoiling events (tertiary structure) and macroscopic anisotropic domains which are parts of a network (quaternary structure) with tunable elasticity up to ∼103 Pa. In contrast, κ-carrageenan in the presence of sodium only produces changes in secondary structure without supercoiling events, prior to formation of gels, highlighting the ion-specificity of the process. Loosely intertwined single helices are observed for ι-carrageenan in the presence of sodium and potassium chloride, providing an elastic mesh with many junction zones, while λ-carrageenan does not undergo any structural change. A generality of the observed behavior may be inferred by extending these observations to a distinct class of polysaccharides, the weak carboxylic polyelectrolyte Gellan gum. These results advance our understanding of ion-specific structural changes of polysaccharides and the physical mechanisms responsible for their gelation.

Incorporation of ß-glucans in meat emulsions through an optimal mixture modeling systems.

The effects of ß-glucans (ßG) in beef emulsions with carrageenan and starch were evaluated using an optimal mixture modeling system. The best mathematical models to describe the cooking loss, color, and textural profile analysis (TPA) were selected and optimized. The cubic models were better to describe the cooking loss, color, and TPA parameters, with the exception of springiness. Emulsions with greater levels of ßG and starch had less cooking loss (<1%), intermediate L* (>54 and <62), and greater hardness, cohesiveness and springiness values. Subsequently, during the optimization phase, the use of carrageenan was eliminated. The optimized emulsion contained 3.13 ±â€¯0.11% ßG, which could cover the intake daily of ßG recommendations. However, the hardness of the optimized emulsion was greater (60,224 ±â€¯1025 N) than expected. The optimized emulsion had a homogeneous structure and normal thermal behavior by DSC and allowed for the manufacture of products with high amounts of ßG and desired functional attributes.

Supramolecular chiral self-assembly and supercoiling behavior of carrageenans at varying salt conditions.

The self-assembly of anionic kappa and iota carrageenan polysaccharides in the presence of NaCl, KCl and CaCl2 is studied by high-resolution atomic force microscopy (AFM). A hierarchical supramolecular chirality amplification over various length scales is observed upon the addition of KCl, whereas in the presence of NaCl and CaCl2 the chains undergo solely a coil-helix transition with stiff kappa carrageenan and more flexible iota carrageenan helical conformations.

Anomalous stiffening and ion-induced coil-helix transition of carrageenans under monovalent salt conditions.

The macromolecular conformations of anionic polysaccharides with decreasing linear charge densities­lambda, iota, and kappa carrageenan­, at varying NaCl concentrations, are studied by single-chain statistical analysis of high-resolution atomic force microscopy (AFM) images. Lambda remains in the random coil conformation, whereas iota and kappa undergo ion-induced coil-helix transitions, with a 2-3-fold increase in chain rigidity. At low ionic strengths, I, the polymer chains sequester Na⁺, leading to a greater flexibility, and beyond a critical I to the formation of an intramolecular single helix. The persistence length exhibits a sublinear dependence on the Debye screening length, κ⁻¹, L(p)(e) ∼ κ(-y) (with 0 < y < 1), deviating from the classical polyelectrolyte behavior expressed by Odijk-Skolnick-Fixman or Barrat-Joanny models. Above a certain I, the L(p) shows an upturn, resulting in polymer stiffening and nonmonotonic behavior. This phenomenon is inferred from specific ion-polymer interactions and/or nonlinear electrostatic physics involving ion-ion correlations.

Development and characterization of carrageenan/grapefruit seed extract composite films for active packaging.

Carrageenan-based antimicrobial films were developed by incorporation of grape fruit seed extract (GSE) at different concentration into the polymer using a solvent casing method and their physical, mechanical, and antimicrobial properties were examined. The carrageenan/GSE composite films appeared yellowish tint due to the polyphenolic compounds in the GSE. SEM analysis showed rough surface with sponge like structures on the cross section of the films. FT-IR results indicated at GSE had good compatibility with carrageenan. The amorphous structure of polymer films was not changed by the incorporation of GSE. But, the addition of GSE increased moisture content, water vapor permeability, and surface hydrophilicity of the films. The tensile strength and elastic modulus decreased with increasing content of GSE, however, the elongation at break increased significantly up to 6.6µg/mL of GSE then decreased thereafter. Thermal stability of the films was not influenced by GSE incorporation. The carrageenan/GSE composite films exhibited great antibacterial activity against food borne pathogens. These results suggest that the carrageenan-based composite films have a high potential for being used as an antimicrobial or active food packaging applications.

Polysaccharide hydrogels with tunable stiffness and provasculogenic properties via α-helix to ß-sheet switch in secondary structure.

Mechanical aspects of the cellular environment can influence cell function, and in this context hydrogels can serve as an instructive matrix. Here we report that physicochemical properties of hydrogels derived from polysaccharides (agarose, κ-carrageenan) having an α-helical backbone can be tailored by inducing a switch in the secondary structure from α-helix to ß-sheet through carboxylation. This enables the gel modulus to be tuned over four orders of magnitude (G' 6 Pa-3.6 × 10(4) Pa) independently of polymer concentration and molecular weight. Using carboxylated agarose gels as a screening platform, we demonstrate that soft-carboxylated agarose provides a unique environment for the polarization of endothelial cells in the presence of soluble and bound signals, which notably does not occur in fibrin and collagen gels. Furthermore, endothelial cells organize into freestanding lumens over 100 µm in length. The finding that a biomaterial can modulate soluble and bound signals provides impetus for exploring mechanobiology paradigms in regenerative therapies.

Ultrasound assisted cold gelation of kappa carrageenan dispersions.

Usually to prepare a gel from kappa carrageenan, an aqueous dispersion of carrageenan powder is heated. This research presents evidence of cold gelation of carrageenan dispersions while no heat is needed. The occurrence of cold gelation is investigated in the absence or presence of potassium ions during ultrasound treatment by studying the mechanical and microstructural properties of the prepared gels using a texture analyzer and scanning electron microscopy, respectively. Carrageenan gels were obtained by applying power ultrasound. By increasing sonication time, gel hardness increased up to a certain level (in absence or presence of K(+) during ultrasound treatment) and further ultrasound application had a negative effect. Moreover, the mechanical properties of the gels in which K(+) ions were added before ultrasound were weaker than the samples when K(+) ions were added after sonication. Relaxation times of gels were calculated using the generalized Maxwell model with three elements.

One-pot synthesis of fluorescent polysaccharides: adenine grafted agarose and carrageenan.

New fluorescent polysaccharides were synthesized by grafting the nucleobase adenine on to the backbones of agarose and κ-carrageenan, which were characterized by FT-IR, (13)C NMR, TGA, XRD, UV, and fluorescence properties. The synthesis involved a rapid water based potassium persulfate (KPS) initiated method under microwave irradiation. The emission spectra of adenine grafted agarose and κ-carrageenan were recorded in aqueous (5×10(-5) M) solution, exhibiting λ(em,max) 347 nm by excitation at 261 nm, affording ca. 30% and 40% enhanced emission intensities, respectively compared to that of pure adenine solution in the same concentration. Similar emission intensity was recorded in the pure adenine solution at its molar equivalent concentrations present in the 5×10(-5) M solution of the agarose and carrageenan grafted products, that is, 3.28×10(-5) M and 4.5×10(-5) M respectively. These fluorescent adenine grafted products may have potential utility in various sensor applications.

Physical state of kappa-carrageenan modulates the mode of action of kappa-carrageenase from Pseudoalteromonas carrageenovora.

Pseudoalteromonas carrageenovora kappa-carrageenase is a glycoside hydrolase involved in the bioconversion of carrageenans. Carrageenans are sulfated galactans that are densely packed in red algal cell walls. Previous crystallographic investigations revealed that the active site of kappa-carrageenase has a tunnel-shaped topology, suggesting a processive mode of action for this enzyme. To biochemically characterize the enzymatic depolymerization of kappa-carrageenan, soluble and solid substrates (in both gel and powder forms) were incubated with P. carrageenovora kappa-carrageenase. The average molecular mass of soluble carrageenan decreased rapidly, and all possible degradation products were observed, suggesting random degradation of kappa-carrageenan. In contrast, as expected for a processive-type carrageenase, the average molecular mass of solid carrageenan decreased very slowly, and tetrasaccharide production was high. Interestingly, experimentally determined processivity was similar for gel and powder, suggesting that, in addition to an adapted catalytic site, the substrate must be in the solid state for kappa-carrageenase processivity to operate, whatever the level of carrageenan ordering.

Preparation and properties of carrageenan microspheres containing allopurinol and local anesthetic agents for the treatment of oral mucositis.

For the treatment of oral mucositis, carrageenan microspheres containing allopurinol and local anesthetic agents, such as lidocaine hydrochloride, dibucaine hydrochloride and tetracaine hydrochloride were prepared using a spray-drying method. As base materials, kappa-carrageenan and iota-carrageenan were evaluated, since carrageenan mitigates bitter taste of lidocaine hydrochloride, dibucaine hydrochloride and tetracaine hydrochloride. The microspheres were spherical and their average diameters were about 10 microm. The drug loading efficiency was more than 70%. Allopurinol and local anesthetic agents became amorphous by the spray drying. Allopurinol and the local anesthetic agents were released from the microspheres for at least 400 min when iota-carrageenan was used as a base material. On the other hand, the release was prolonged to 600 min when kappa-carrageenan was used. The microspheres were spread and made membranes at the air/water interfaces immediately after dropped on the water surfaces. The properties of the microspheres such as dispersing efficacy and membrane production on the water surfaces suggest that the microspheres can uniformly cover inner surfaces of oral cavity to prevent and treat oral mucositis.

Tensile and tear strength of carrageenan film from Philippine eucheuma species.

The tensile and tear strength of carrageenan film from Philippines Eucheuma species were investigated using NEC tensilon universal-testing machine according to American Society for Testing Materials methods. These properties are important for assessing carrageenan film as packaging material. The kappa and iota types were used in the study. The effect of glycerine on the tensile and tear strength including elongation was also evaluated. Addition of glycerine tended to lower the tensile strength of the film and increase its elongation properties including the tear strength. Carrageenan film without glycerine was much stronger. Glycerine made the film more flexible and easy to deform. The composite film of carrageenan and konjac gum did not exhibit elongation. It also showed higher tensile strength than did the composite film of carrageenan and xanthan gum. Compared with iota-type carrageenan film, kappa-type carrageenan film without glycerine was more comparable to low-density polyethylene (LDPE) film in terms of tensile strength as was the composite film of carrageenan-konjac gum. The kappa-type carrageenan film with glycerine was more comparable to LDPE film in terms of tear strength. The elongation reading for carrageenan film was lower than that for LDPE film. Morphologic studies showed that the carrageenan film had sets of pores distributed randomly at different places as compared to LDPE film. It also showed that the carrageenan film was more fibrous than LDPE film.

Evaluation of bioadhesive glipizide spheres and compacts from spheres prepared by extruder/marumerizer technique.

The objective of this study was to attempt to deliver glipizide from spheres and compacts containing the natural polymer Carrageenan (Gelcarin, GP 812) and prepared by extruder/marumerizer technique. A second objective was to evaluate the mucoadhesive strength of the bioadhesive spheres onto the mucus membrane of rabbit. The effects of polymer, drug level, and type of spheronizing material were evaluated. All sphere formulations were compacted into tablets using a rotary Manesty B-3B machine equipped with 12/32 flat face tooling. Results show drug release from spheres and compacts decreased as the level of Carrageenan was increased. However as the level of drug was increased drug release also increased. Spheres containing Avicel PH-101 gave higher drug release than spheres of the same composition but prepared with Avicel RC-581. In general, the drug release from tablets was higher than its corresponding spheres and drug release from spheres and tablets containing Carrageenan was higher than control spheres and tablets of the same composition but without Carrageenan. Tablet formulations compacted from spheres containing Avicel RC-581 gave higher release rate constants than tablet formulations of the same composition but prepared with Avicel PH-101. The bioadhesion study showed that mucoadhesion strength between spheres and mucus membrane of the rabbit depends on the levels of polymer, drug, and type of spheronizing material. Developed bioadhesive spheres and tablets increase the solubility of glipizide which may increase its bioavailability and also increased the adherence of the bioadhesive systems to the mucous membrane so that once daily dose can be administered.

Microstructure of aggregated and nonaggregated kappa-carrageenan helices visualized by atomic force microscopy.

Gelation of kappa-carrageenan is believed to involve two steps: helix formation on cooling and a further specific cation (salt) induced side-by-side aggregation of helices. Samples that should contain aggregated and also nonaggregated "helices" of kappa-carrageenan were prepared in aqueous solutions, spread onto freshly cleaved mica surfaces, and visualized under butanol using atomic force microscopy. In the presence of an excessive amount of a gel-promoting salt, KCl, kappa-carrageenan appeared to form rigid rodlike structures considered as large aggregates of double helices. Even when the side-by-side interhelical aggregation was suppressed by diluting random coiled solutions prior to cooling, by adding an aggregation-impeding salt, NaI, or by transforming kappa-carrageenan into the tetramethylammonium (TMA) salt, branched rodlike structures were still evident, suggesting that the side-by-side aggregation of helices is not a prerequisite for kappa-carrageenan to form a network structure, at least locally. Even in the absence of factors that promote side-by-side aggregation, kappa-carrageenan helices appeared to be capable of associating and forming gel networks.

Imaging of carrageenan macrocycles and amylose using noncontact atomic force microscopy.

Samples of kappa-carrageenan, iota-carrageenan, and synthetic amylose have been examined by atomic force microscopy (AFM). All samples were spray deposited from aqueous solutions onto freshly cleaved mica, air dried, and imaged in air using noncontact atomic force microscopy (NCAFM). Images of single stranded amylose and carrageenan are presented. At relatively low polymer concentrations in the presence of NaCl iota-carrageenan formed circles that appear to be predominantly head-to-tail associated unimeric duplex (double stranded) structures. At higher iota-carrageenan concentrations the polymer forms circles and aggregates that appear to involve dimeric duplex structure. Direct comparison of synthetic amylose molecular weights determined from NCAFM images with results from solution measurements showed that NCAFM provides an excellent way to measure amylose molecular weight and molecular weight distribution. It is shown that synthetic amylose is single stranded in aqueous solution and that the chain length distribution is broader than the Poisson distribution anticipated from polymerization theory.