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Quick drying after water or sweat absorption is an important function of underwear. In this study, the hydroxy groups of cotton knitted fabrics (CFs) were partially acetylated, maintaining the original fabric structure. The following three heterogeneous acetylation processes were used: Ac-I (Ac2O/H2SO4/toluene), Ac-II (Ac2O/H2SO4/AcOH/water), and Ac-III (Ac2O/AcONa) systems (Ac2O, acetic anhydride; AcOH, acetic acid; AcONa, sodium acetate). Acetylated cotton knitted fabrics (AcCFs) with degree of substitution (DS) ≤ 0.5 and yields of > 80% were prepared. AcCFs prepared with the Ac-III system gave high degree of polymerization (DP) values of > 1500, whereas those prepared with the Ac-II system exhibited low DP values of ≤ 400. The moisture contents of AcCFs at 20 °C and 65% relative humidity decreased from 7.1 to 4.7% with increasing DS value up to 0.46; introducing hydrophobic acetyl groups into the CFs decreased their hydrophilic nature. Quick drying similar to that of a polyester fabric was achieved for some of the AcCFs with DS values of < 0.2. When the acetyl groups in the AcCFs were homogeneously distributed across each fiber width (achieved for AcCFs prepared with the Ac-II system), quick drying was evident in the AcCFs. The crystallinities and crystal widths of cellulose I for the AcCFs with DS values of ≤ 0.28 were almost unchanged compared with those of the original CFs. However, neither the crystallinities nor crystal widths of cellulose I were directly related to quick drying after water absorption. Thermal degradation of the AcCFs varied between the acetylation systems, and depended on the DP values and/or the presence of sulfate ester groups in the AcCFs. [ABSTRACT FROM AUTHOR]

Chitosan-based materials have broad applications, from biotechnology to pharmaceutics. Recent experiments showed that the degree and pattern of acetylation along the chitosan chain modulate its biological and physicochemical properties; however, the molecular mechanism remains unknown. Here, we report, to the best of our knowledge, the first de novo all-atom molecular dynamics (MD) simulations to investigate chitosan's self-assembly process at different degrees and patterns of acetylation. Simulations revealed that 10 mer chitosan chains with 50% acetylation in either block or alternating patterns associate to form ordered nanofibrils comprised of mainly antiparallel chains in agreement with the fiber diffraction data of deacetylated chitosan. Surprisingly, regardless of the acetylation pattern, the same intermolecular hydrogen bonds mediate fibril sheet formation while water-mediated interactions stabilize sheet–sheet stacking. Moreover, acetylated units are involved in forming strong intermolecular hydrogen bonds (NH–O6 and O6H–O7), which offers an explanation for the experimental observation that increased acetylation lowers chitosan's solubility. Taken together, the present study provides atomic-level understanding the role of acetylation plays in modulating chitosan's physiochemical properties, contributing to the rational design of chitosan-based materials with the ability to tune by its degree and pattern of acetylation. Additionally, we disseminate the improved molecular mechanics parameters that can be applied in MD studies to further understand chitosan-based materials. [ABSTRACT FROM AUTHOR]

Pectin was extracted from coffee pulp using 0.1 M H2SO4. The Box Behnken Design based Response surface methodology was applied to optimize pectin yield. The impact of extraction time (45–75 min), temperature (80–100 °C), solid to liquid ratio (SLR) (1:20, 1:27.5, and 1:35), and pH (1.5, 2, and 2.5) on pectin yield were studied. Under optimal extraction conditions (84 °C, 75 min, SLR of 1:20 and pH: 1.5), physical, chemical, structural and antioxidant properties of pectin were examined. The results of the physicochemical analysis are: acetyl value: 1.10 ± 0.05 %, equivalent weight: 1429 ± 54 g/mol, anhydrouronic acid: 57.1 ± 0.9 %, degree of esterification: 78.5 ± 1.8 %, moisture content: 8.5 ± 1.5 % and ash content: 4.3 ± 0.9 %. FTIR analysis indicated the (-OH) peak of pectin was lower and shifted left compared to treated and untreated coffee pulp powder. SEM analysis shows a smoother surface, whereas XRD shows a less amorphous structure of pectin. The total phenolic and flavonoid content of coffee pulp pectin was found to be 26.7 μg Gallic Acid Equivalent/mg and 0.8957 μg Quercetin Equivalent/mg, respectively. Antioxidant analysis showed significant antioxidant properties (IC50 = 642.31 ± 30.43 μg/mL). The predicted and actual pectin yields at the optimal extraction condition were 14.39 and 13.7 %, respectively, with R2 = 0.95 that indicate the model can represent the experiment. Therefore, achieving a maximum pectin yield with improved antioxidant and other physicochemical qualities ensures that coffee pulp can potentially serve as a viable commercial source of pectin.

Aloe vera (Aloe barbadensis Miller) gel is a frequently used ingredient in many food pro-ducts, particularly beverages, due to its reported health benefits. Studies have identified acemannan, a polysaccharide rich in mannose units which are partially or fully acetylated, as the primary bioactive compound in Aloe vera gel. The acemannan content and its degree of acetylation (DA) were measured in 15 different commercial beverages containing Aloe vera at varying concentrations (from 30% to 99.8%) as listed on the label. Other biopolymers such as pectins, hemicelluloses, and cellulose were also evaluated. Flavoured beverages (seven samples labelled as containing from 30% to 77% Aloe vera) presented low levels of acemannan (<30 mg/100 g of fresh sample) and were fully deacetylated in most cases. These samples had high levels of other polymers such as pectins, hemicelluloses, and cellulose, likely due to the addition of fruit juices for flavour. Unflavoured beverages (eight samples, with Aloe vera concentrations above 99% according to their labels) had variable levels of acemannan, with only three containing more than 160 mg/100 g of fresh sample. In fact, four samples had less than 35 mg acemannan/100 g of fresh sample. DA levels in all but one sample were lower than 35%, possibly due to processing techniques such as pasteurization causing degradation and deacetylation of the acemannan polymer. Legislation regarding Aloe vera products is limited, and manufacturers are not required to disclose the presence or quality of bioactive compounds in their products, leaving consumers uncertain about the true properties of the products they purchase. [ABSTRACT FROM AUTHOR]

Chitosan-based materials have broad applications, from biotechnology to pharmaceutics. Recent experiments showed that the degree and pattern of acetylation along the chitosan chain modulate its biological and physicochemical properties; however, the molecular mechanism remains unknown. Here, we report, to the best of our knowledge, the first de novo all-atom molecular dynamics (MD) simulations to investigate chitosan’s self-assembly process at different degrees and patterns of acetylation. Simulations revealed that 10 mer chitosan chains with 50% acetylation in either block or alternating patterns associate to form ordered nanofibrils comprised of mainly antiparallel chains in agreement with the fiber diffraction data of deacetylated chitosan. Surprisingly, regardless of the acetylation pattern, the same intermolecular hydrogen bonds mediate fibril sheet formation while water-mediated interactions stabilize sheet–sheet stacking. Moreover, acetylated units are involved in forming strong intermolecular hydrogen bonds (NH–O6 and O6H–O7), which offers an explanation for the experimental observation that increased acetylation lowers chitosan’s solubility. Taken together, the present study provides atomic-level understanding the role of acetylation plays in modulating chitosan’s physiochemical properties, contributing to the rational design of chitosan-based materials with the ability to tune by its degree and pattern of acetylation. Additionally, we disseminate the improved molecular mechanics parameters that can be applied in MD studies to further understand chitosan-based materials.

In this study, the physicochemical, rheological, and functional properties of colloidal chitin prepared from commercial shrimp shell α-chitin by acid hydrolysis at ambient temperature for various treatment durations were investigated. With increasing treatment duration, the functional properties viz solubility, water and fat binding capacity, and emulsion capacity increased significantly (p0.05) among treatment durations. The degree of deacetylation decreased as treatment duration increased (p

Aloe vera (Aloe barbadensis Miller) gel is a frequently used ingredient in many food pro-ducts, particularly beverages, due to its reported health benefits. Studies have identified acemannan, a polysaccharide rich in mannose units which are partially or fully acetylated, as the primary bioactive compound in Aloe vera gel. The acemannan content and its degree of acetylation (DA) were measured in 15 different commercial beverages containing Aloe vera at varying concentrations (from 30% to 99.8%) as listed on the label. Other biopolymers such as pectins, hemicelluloses, and cellulose were also evaluated. Flavoured beverages (seven samples labelled as containing from 30% to 77% Aloe vera) presented low levels of acemannan (

Endotoxemia is mainly caused by a massive burst of inflammatory cytokines as a result of lipopolysaccharide (LPS) invasion. Chitooligosaccharides (COS) is expected to be a potential drug for relieving endotoxemia due to its anti-inflammatory properties. However, the structural parameters of COS are often ambiguous, and the effect of degree of acetylation (DA) of COS on its anti-inflammatory remains unknown. In this study, four COSs with different DAs (0%, 12%, 50% and 85%) and the same oligomers distribution were successfully obtained. Their structures were confirmed by 1H NMR and MS analysis. Then, the effect of DA on the anti-inflammatory activity and relieving endotoxemia potential of COS was researched. The results revealed that COS with a DA of 12% had better anti-inflammatory activity than COSs with other DAs, mainly in inhibiting LPS-induced inflammatory cytokines burst, down-regulating its mRNA expression and reducing phosphorylation of IκBα. Furthermore, this COS showed an obviously protective effect on endotoxemia mice, such as inhibiting the increase in inflammatory cytokines and transaminases, alleviating the injury of liver and intestinal tissue. This study explored the effect of DA on the anti-inflammatory activity of COS for the first time and lays the foundation for the development of COS as an anti-inflammatory drug against endotoxemia. [ABSTRACT FROM AUTHOR]

Chitin is the second most abundant bio-polymer after cellulose in earth. This carbohydrate polymer is one of the major components of the cell wall of some micro-organisms and exoskeleton of certain invertebrates. Recent studies show the potential application of chitin and its derivatives in chemical, medical, agricultural, food technology, pollutant treatment and textile sectors. The traditional resources of chitin for industrial processing are crustacean shells and fungal mycelia. This work attempts to extract and characterize chitin from the edible insect Coridius nepalensis. The results were compared with the commercial chitin extracted from shrimp (Pandalus borealis). The chitin yield of from C. nepalensis was found to be 43.97% of its dry weight. The degree of acetylation was recorded as 57.67%. FT-IR spectrum showed the extracted chitin as α-allomorph. The peaks of the chitin extracted from C. nepalensis in X-ray diffractogram are less sharp compared to commercial shrimp chitin. Thus, the degree of acetylation, FT-IR spectrum and XRD show mixture of chitin and chitosan in the extracted chitin from C. nepalensis. The crystalline index (CrI110) for C. nepalensis and commercial shrimp chitin was calculated as 86.33% and 71.56% respectively. Scanning electron micrographs indicated granular surface morphology the extracted chitin of C. nepalensis, in contrast to commercial shrimp chitin, which was fibrous and porous. Thus, the present study indicates the possibility to use the adults of C. nepalensis as a new and alternative source of biomaterial. [ABSTRACT FROM AUTHOR]

This study investigates the impact of water and salinity stress on Aloe vera , focusing on the role of Aloe vera polysaccharides in mitigating these stresses. Pectins and acemannan were the most affected polymers. Low soil moisture and high salinity (NaCl 80 mM) increased pectic substances, altering rhamnogalacturonan type I in Aloe vera gel. Aloe vera pectins maintained a consistent 60 % methyl-esterification regardless of conditions. Interestingly, acemannan content rose with salinity, particularly under low moisture, accompanied by 90 to 150 % acetylation increase. These changes improved the functionality of Aloe vera polysaccharides: pectins increased cell wall reinforcement and interactions, while highly acetylated acemannan retained water for sustained plant functions. This study highlights the crucial role of Aloe vera polysaccharides in enhancing plant resilience to water and salinity stress, leading to improved functional properties. [Display omitted] • The role of acemannan and pectin on the tolerance of Aloe vera to saline-water stress was studied. • Acemannan and pectic polysaccharides levels increase with higher saline-water stress. • Abiotic stress leads to increased acetylation of acemannan and branching of pectic polysaccharides. • Highly acetylated acemannan may improve intracellular water management during saline-water stress. • The presence of highly branched pectin as a result of abiotic stress could reinforce the cell wall. [ABSTRACT FROM AUTHOR]

Endotoxemia is mainly caused by a massive burst of inflammatory cytokines as a result of lipopolysaccharide (LPS) invasion. Chitooligosaccharides (COS) is expected to be a potential drug for relieving endotoxemia due to its anti-inflammatory properties. However, the structural parameters of COS are often ambiguous, and the effect of degree of acetylation (DA) of COS on its anti-inflammatory remains unknown. In this study, four COSs with different DAs (0%, 12%, 50% and 85%) and the same oligomers distribution were successfully obtained. Their structures were confirmed by 1H NMR and MS analysis. Then, the effect of DA on the anti-inflammatory activity and relieving endotoxemia potential of COS was researched. The results revealed that COS with a DA of 12% had better anti-inflammatory activity than COSs with other DAs, mainly in inhibiting LPS-induced inflammatory cytokines burst, down-regulating its mRNA expression and reducing phosphorylation of IκBα. Furthermore, this COS showed an obviously protective effect on endotoxemia mice, such as inhibiting the increase in inflammatory cytokines and transaminases, alleviating the injury of liver and intestinal tissue. This study explored the effect of DA on the anti-inflammatory activity of COS for the first time and lays the foundation for the development of COS as an anti-inflammatory drug against endotoxemia.

The esterification of galacturonic acid with methanol and/or acetic acid is important for the structural analysis of pectin. Although several methods have been reported for determining the degree of methylesterification (DM) and acetylation (DAc), the present study compares and optimizes three methods (titration, FT-IR and HPLC) using commercial citrus pectin (CP). Our results showed that the DM of CP was 47.0%, 47.6% and 48.0% as determined by titration, FT-IR and HPLC, respectively, suggesting that DM determination is nearly identical using any of these methods. However, the titration approach requires more sample than the other two. HPLC showed that the DAc of CP was 1.6%, an approach that can be used to determine the DM and DAc of pectin simultaneously. Here, we simplified and optimized sample treatment for HPLC analysis and compared it with the reported literature. Our results provide useful information for choosing appropriate methods for determining the DM and DAc of pectin based on various sample properties and experimental conditions. [ABSTRACT FROM AUTHOR]

Pectin was extracted from coffee pulp using 0.1 M H2SO4. The Box Behnken Design based Response surface methodology was applied to optimize pectin yield. The impact of extraction time (45-75 min), temperature (80-100 °C), solid to liquid ratio (SLR) (1:20, 1:27.5, and 1:35), and pH (1.5, 2, and 2.5) on pectin yield were studied. Under optimal extraction conditions (84 °C, 75 min, SLR of 1:20 and pH: 1.5), physical, chemical, structural and antioxidant properties of pectin were examined. The results of the physicochemical analysis are: acetyl value: 1.10 ± 0.05 %, equivalent weight: 1429 ± 54 g/mol, anhydrouronic acid: 57.1 ± 0.9 %, degree of esterification: 78.5 ± 1.8 %, moisture content: 8.5 ± 1.5 % and ash content: 4.3 ± 0.9 %. FTIR analysis indicated the (-OH) peak of pectin was lower and shifted left compared to treated and untreated coffee pulp powder. SEM analysis shows a smoother surface, whereas XRD shows a less amorphous structure of pectin. The total phenolic and flavonoid content of coffee pulp pectin was found to be 26.7 μg Gallic Acid Equivalent/mg and 0.8957 μg Quercetin Equivalent/mg, respectively. Antioxidant analysis showed significant antioxidant properties (IC50 = 642.31 ± 30.43 μg/mL). The predicted and actual pectin yields at the optimal extraction condition were 14.39 and 13.7 %, respectively, with R 2  = 0.95 that indicate the model can represent the experiment. Therefore, achieving a maximum pectin yield with improved antioxidant and other physicochemical qualities ensures that coffee pulp can potentially serve as a viable commercial source of pectin., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors.)

Cryptococcus neoformans is an opportunistic fungal pathogen that infects ~280,000 people every year, causing >180,000 deaths. The human immune system recognizes chitin as one of the major cell-wall components of invading fungi, but C. neoformans can circumvent this immunosurveillance mechanism by instead exposing chitosan, the partly or fully deacetylated form of chitin. The natural production of chitosans involves the sequential action of chitin synthases (CHSs) and chitin deacetylases (CDAs). C. neoformans expresses four putative CDAs, three of which have been confirmed as functional enzymes that act on chitin in the cell wall. The fourth (CnCda4/Fpd1) is a secreted enzyme with exceptional specificity for D-glucosamine at its -1 subsite, thus preferring chitosan over chitin as a substrate. We used site-specific mutagenesis to reduce the subsite specificity of CnCda4 by converting an atypical isoleucine residue in a flexible loop region to the bulkier or charged residues tyrosine, histidine, and glutamic acid. We also investigated the effect of CnCda4 deacetylation products on human peripheral blood-derived macrophages, leading to a model explaining the function of CnCda4 during infection. We propose that CnCda4 is used for the further deacetylation of chitosans already exposed on the C. neoformans cell wall (originally produced by CnChs3 and CnCda1 to 3) or released from the cell wall as elicitors by human chitinases, thus making the fungus less susceptible to host immunosurveillance. The absence of CnCda4 during infection could therefore promote the faster recognition and elimination of this pathogen. [ABSTRACT FROM AUTHOR]

Chitin is a structural polysaccharide that is found in crustaceans, insects, fungi and some yeasts. Chitin deacetylation produces chitosan, a well-studied biopolymer with reported chemical and biological properties for diverse potential applications for drug delivery, metal ion absorption, scaffolds and tissue engineering. Most known properties of chitosan have been determined from samples obtained from a variety of sources and in different conditions, this is, from chitosans with a wide range of degrees of N-acetylation (DA) and molecular weight (MW). However, as for any copolymer, the physicochemical and mechanical characteristics of chitosan highly depend on their monomer composition (DA) and chain size (MW). This work presents a simple methodology to produce chitosans with specific and predictive DA and MW. Reaction with acetic anhydride proved to be an efficient method to control the acetylation of chitosan, DAs between 10.6% and 50.6% were reproducibly obtained. In addition to this, MWs of chitosan chains were reduced in a controlled manner in two ways, by ultrasound and by acidic hydrolysis at different temperatures, samples with MWs between 130 kDa and 1300 kDa were obtained. DAs were determined by 1H-NMR and MWs by gel permeation chromatography.

The esterification of galacturonic acid with methanol and/or acetic acid is important for the structural analysis of pectin. Although several methods have been reported for determining the degree of methylesterification (DM) and acetylation (DAc), the present study compares and optimizes three methods (titration, FT-IR and HPLC) using commercial citrus pectin (CP). Our results showed that the DM of CP was 47.0%, 47.6% and 48.0% as determined by titration, FT-IR and HPLC, respectively, suggesting that DM determination is nearly identical using any of these methods. However, the titration approach requires more sample than the other two. HPLC showed that the DAc of CP was 1.6%, an approach that can be used to determine the DM and DAc of pectin simultaneously. Here, we simplified and optimized sample treatment for HPLC analysis and compared it with the reported literature. Our results provide useful information for choosing appropriate methods for determining the DM and DAc of pectin based on various sample properties and experimental conditions.

Mealworm (Tenebrio molitor L.) is a classic edible insect with high nutritional value for substituting meats from vertebrates. While interest in mealworms has increased, the determination of carbohydrate constituents of mealworms has been overlooked. Thus, the aim of the present study was to investigate the carbohydrate content and composition of mealworms. In addition, the characteristics of mealworm chitin were determined as these were the major components of mealworm carbohydrate. The crude carbohydrate content of mealworms was 11.5%, but the total soluble sugar content was only 30% of the total carbohydrate content, and fructose was identified as the most abundant free sugar in mealworms. Chitin derivatives were the key components of mealworm carbohydrate with a yield of 4.7%. In the scanning electron microscopy images, a lamellar structure with α-chitin configuration was observed, and mealworm chitosan showed multiple pores on its surface. The overall physical characteristics of mealworm chitin and chitosan were similar to those of the commercial products derived from crustaceans. However, mealworm chitin showed a significantly softer texture than crustacean chitin with superior anti-inflammatory effects. Hence, mealworm chitin and chitosan could be employed as novel resources with unique advantages in industries.

We report on a method for the preparation of cellulose/chitin composite materials from the ionic liquid 1-butyl-3-methylimidazolium acetate and γ-valerolactone as a biosourced sustainable co-solvent. Element analysis and attentuated total reflectance Fourier transform infrared spectroscopy show that the average degree of acetylation of chitin in the composite materials was around 82.5%. This indicates that chitin is not deacetylated to chitosan during the dissolution process. The X-ray diffraction results show that the degree of crystallinity of the composite materials increases from amorphous to 59% with increasing chitin concentration accompanied by a developing crystallite size up to 3 nm. Mechanical testing yielded a maximum tensile stress of 4.7 MPa, an elastic modulus of 27.4 MPa and a breaking elongation of 78.7% for the composites with 80 wt% chitin. In addition, water contact angle measurements indicated that the presence of chitin rendered the materials more hydrophobic. [ABSTRACT FROM AUTHOR]

Acetyl groups are backbone outreaches that enhance inter-fibril connection in chitin and chitosan fibril bundle. Removal of acetyl groups affects flexibility of chitosan fibril bundle, thereby affecting mechanical strength of chitosan-based products. Understandings of relationship between degree of acetylation and flexibility of chitin fibril bundle conduce to optimization of synthetic chitin materials. Here, the relationship is examined by performing molecular dynamics simulations. Coiling of chitin and chitosan fibril bundle with different degree of acetylation is observed and flexibility of fibrils is measured. Number and alignment of acetyl groups are found to be important factors determining the flexibility of chitin and chitosan fibril bundle. Structural instability can be caused by incompatible alignment of acetyl groups. Our findings on synthetic chitin-based materials indicate that adding a small amount of acetyl groups to chitosan can significantly enhance the integrity of fibril bundle. [ABSTRACT FROM AUTHOR]

In the present study, a well-defined set of chitosans, with different degrees of acetylation (DA) and degrees of polymerization (DP), were processed by solution electrospraying from a water-based solvent. The solution properties, in terms of surface tension, conductivity, viscosity, and pH, were characterized and related to the physico-chemical properties of the chitosans. It was observed that both DA and DP values of a given chitosan, in combination with biopolymer concentration, mainly determined solution viscosity. This was, in turn, the major driving factor that defined the electrosprayability of chitosan. In addition, the physico-chemical properties of chitosans highly influenced solution conductivity and results indicated that the chitosan solutions with low or low-to-medium values of conductivity were the most optimal for electrospraying. The results obtained here also demonstrate that a good process control can be achieved by adjusting the working conditions,i.e. applied voltage, flow-rate, and tip-to-collector distance. Finally, it was also shown that electrosprayability of chitosan with inadequate physico-chemical properties can be improved by solution mixing of very different kinds of this polysaccharide. The resultant electrosprayed submicron chitosan capsules can be applied for encapsulation of food additives and to develop bioactive coatings of interest in food packaging, where these particles alone or containing functional ingredients can be released from the package into the food to promote a health benefit. [ABSTRACT FROM AUTHOR]

Peripheral nerve injuries have produced major concerns in regenerative medicine for several years, as the recovery of normal nerve function continues to be a significant clinical challenge. Chitosan (CHT), because of its good biocompatibility, biodegradability and physicochemical properties, has been widely used as a biomaterial in tissue engineering scaffolding. In this study, CHT membranes were produced with three different Degrees of Acetylation (DA), envisioning its application in peripheral nerve regeneration. The three CHT membranes (DA I: 1%, DA II: 2%, DA III: 5%) were extensively characterized and were found to have a smooth and flat surface, with DA III membrane having slightly higher roughness and surface energy. All the membranes presented suitable mechanical properties and did not show any signs of calcification after SBF test. Biodegradability was similar for all samples, and adequate to physically support neurite outgrowth. The in vitro cell culture results indicate selective cell adhesion. The CHT membranes favoured Schwann cells invasion and proliferation, with a display of appropriate cytoskeletal morphology. At the same time they presented low fibroblast infiltration. This fact may be greatly beneficial for the prevention of fibrotic tissue formation, a common phenomenon impairing peripheral nerve regeneration. The great deal of results obtained during this work permitted to select the formulation with the greatest potential for further biological tests. [ABSTRACT FROM AUTHOR]

The rising demand for chitin and chitosan in chemical, agro-food, and healthcare industries is creating a need for rapid and high-throughput analysis. The physicochemical properties of these biopolymers are greatly dependent on the degree of acetylation (DA). Conventional methods for DA determination, such as LC-MS and 1H NMR, are time-consuming when performed on many samples, and therefore efficient methods are needed. Here, high-throughput microplate-based FTIR and FT-Raman methods were compared with their manual counterparts. Partial least squares regression models were based on 30 samples of chitin and chitosan with reference DA values obtained by LC-MS and 1H NMR, and the models were validated on an independent test set of 16 samples. The overall predictive accuracy of the high-throughput methods was at the same level as the manual methods and the well-established LC-MS and 1H NMR methods. Therefore, high-throughput FTIR and FT-Raman DA determination methods have great potential to serve as fast and economical substitutes for traditional methods. [Display omitted] [ABSTRACT FROM AUTHOR]

The degree of acetylation of chitosan is an important parameter to be taken into consideration when chitosan is used as starting material for realization of derived products with applications in pharmacy and medicine. Although considered a reliable method, the determination of the degree of acetylation of chitosan by UV spectrophotometry has as main drawback: the lack of accuracy. Starting from an already published method, based on the UV spectrophotometry of N-acetylglucosamine and glucosamine molecules and considering that UV absorbance of chitosan is due to the absorbance of the two monomers, some preliminary results of an improved method for estimation of the degree of acetylation of chitosan are proposed. The improvements are based on the use of phosphoric acid as solvent for chitosan (and of monomer molecules - N-acetylglucosamine and glucosamine) and the use of optical density at 197 nm. [ABSTRACT FROM AUTHOR]

The objective of this paper is to emphasize the fact that while consistent interest has been paid to the industrial use of chitosan, minor attention has been devoted to spread the knowledge of a good characterization of its physico-chemical properties. Therefore, the paper attempts to critically comment on the conflicting experimental results, highlighting the facts, the myths and the controversies. The goal is to indicate how to take advantage of chitosan versatility, to learn how to manage its variability and show how to properly tackle some unexpected undesirable features. In the sections of the paper various issues that relate chitosan properties to some basic features and to advanced solutions and applications are presented. The introduction outlines some historical pioneering works, where the chemistry of chitosan was originally explored. Thereafter, particular reference is made to analytical purity, characterization and chain modifications. The macromolecular characterization is mostly related to molecular weight and to degree of acetylation, but also refers to the conformational and rheological properties and solution stability. Then, the antimicrobial activity of chitosan in relation with its solubility is reviewed. A section is dedicated to the formulation of chitosan biomaterials, from gel to nanobeads, exploring their innovative application as active carrier nanoparticles. Finally, the toxicity issue of chitosan as a polymer and as a constructed nanomaterial is briefly commented in the conclusions. [ABSTRACT FROM AUTHOR]

Dietary fiber like konjac glucomannan (KGM) is important in maintaining good human health. There is no established method for quantifying the average degree of acetylation DA of this polysaccharide. Polysaccharides are notoriously difficult to dissolve. In this study, KGM could not be fully dissolved in common solvents and was characterized in the solid state. ATR-FTIR spectroscopy enabled a fast qualitative assessment of acetylation, selective to the outer layer of KGM particles, and identifying excipients like magnesium stearate. Average DA was quantified for the first time with solid-state 13C NMR in KGM: semi-quantitative measurements on the same arbitrary scale by cross polarization (1 to 2 days) were calibrated with a few longer single-pulse excitation measurements (approximately 1 week). DA values ranged from 4 to 8% of the hexoses in the backbone, in agreement with previously reported values. This method could be used for quality control and standardization of KGM products. [Display omitted] [ABSTRACT FROM AUTHOR]

Chitosan is a linear aminopolysaccharide that has been widely used for the formation of chitosan-based nanoparticles by ionic gelation with sodium tripolyphosphate (TPP). Often, the experimental design used to obtain these systems does not take into consideration important variables, such as the degree of acetylation (DA) and the molecular weight ( M w ) of chitosan. In this work, we studied the formation of chitosan-TPP nanoparticles with chitosan samples of varying DA and M w (DA0 ∼ 0–47% and M w ∼ 2.5–282 kDa). We addressed the influence the degree of space occupancy and the degree of crosslinking on the physical properties of chitosan-TPP nanoparticles. Nanoparticles that comprised chitosan of DA ∼ 0–21.7% behaved differently than those made of chitosan of DA ∼ 34.7–47%. We attributed these differences to the polymer conformation and chain flexibility of the distinct chitosans in solution. Moreover, chitosan of high M w were found to have a stronger preference for incorporating into the formed nanoparticles than do low- M w ones, as determined by SEC-HPLC. These results open new perspectives to understand the formation of chitosan nanoparticles by the ionic gelation technique. [ABSTRACT FROM AUTHOR]

Chitin is one of the most abundant polysaccharides on earth. It consists of repeating β-1,4 linked N-acetylated glucosamine (A) units. Chitosan is an N-deacetylated product of chitin. Chitosan and its derivatives have broad medical applications as drugs, nutraceuticals, or drug delivery agents. However, a reliable analytical method for quality control of medically used chitosans is still lacking. In current study, nitrous acid was used to cleave all glucosamine residues in chitosan into 2,5-anhydromannose (M) or M at the reducing end of di-, tri-, and oligosaccharides. PMP, i.e. 1-phenyl-3-methyl-5-pyrazolone, was used to label all the Ms. Online UV detection allowed quantification of all M-containing UV peaks whereas online MS analysis directly identified 11 different kinds of mono-, di-, tri-, and oligosaccharides that correlated each oligosaccharide with specific UV peak after HPLC separation. The DA (degree of acetylation) for chitosans was calculated based on the A/(A+M) value derived from the UV data. This newly developed method had several advantages for quality control of chitosan: 1. the experimental procedures were extensively optimized; 2. the reliability of the method was confirmed by online LC-MS analysis; 3. the DA value was obtainable based on the UV data after HPLC analysis, which was comparableto that of 1 H NMR and conductometric titration analyses; 4. finally and most importantly, this method could be used to obtain the DA as well as chemical acetylation/deacetylation mechanisms for chitosan by any laboratory equipped with a HPLC and an online UV detector. [ABSTRACT FROM AUTHOR]

A novel approach to measure the degree of acetylation in biopolymers applying a combination of Zemplén-deacetylation by sodium methanolate and GC–MS methodology is introduced. The development focuses on very low limits of detection to cover also samples with extremely low degrees of acetylation which hitherto eluded accurate determination. Free acetic acid or inorganic acetates, often present in biopolymer samples, do not disturb the quantification. Two techniques to measure the Zemplén-released methyl acetate were comparatively assessed, direct injection of the liquid phase and a SPME-based approach, the former being more straightforward, but being inferior to the latter in sensitivity. By applying isotopically labeled methyl acetate released from 4- O -( 13 C 2 -acetyl)-vanillin as the internal standard, influences, such as varying moisture contents, are corrected, improving the overall method reliability to a large extent. The combination of Zemplén-release of acetyl groups in biopolymers as methyl acetate, in connection with its accurate quantification by SPME–GC–MS, was found to be the method of choice for routine, yet very accurate analysis of a wide range of acetylation degrees of biopolymers, showing satisfying analytical parameters along with easy handling and widest applicability. Limit of detection for acetylated cellulose samples is 0.09 nmol/mg, for hemicellulose samples 0.48 nmol/mg. [ABSTRACT FROM AUTHOR]

RESUMO: Procedimentos adequados para as purificações de quitosana comercial como cloridrato e na forma neutralizada são descritos. As amostras obtidas apresentam elevado grau de pureza e baixo teor de cinzas conforme verificado através de termogravimetria. As determinações de graus de acetilação por espectroscopia de ressonância magnética nuclear ( ou = 23,6%) e por titulações ( ou = 23,0%) apresentam muito boa concordância. O cloridrato de quitosana é solúvel em água, o que permite determinações de viscosidade intrínseca em solventes aquosos na ausência de excesso de ácido e facilita a comparação de quitosana com outros polieletrólitos quanto a estimativas de rigidez/flexibilidade.ABSTRACT: The procedures for the purification of a commercial chitosan as a hydrochloride and in the neutralized form are described. By applying these procedures, pure chitosans of low ash contents are obtained as determined by thermogravimetry. The agreement between the values of degrees of acetylation determined by conductimetric titrations ( or = 23,0%) and by ¹H nmr spectroscopy ( or = 23,6%) is very good. Chitosan hydrochloride being soluble in pure water allows the determination of intrinsic viscosity in dilute acid-free aqueous solutions and makes easier the comparison of chitosan and other polyelectrolytes with respect to their chain rigidity/flexibility.

The objective of this paper is to emphasize the fact that while consistent interest has been paid to the industrial use of chitosan, minor attention has been devoted to spread the knowledge of a good characterization of its physico-chemical properties. Therefore, the paper attempts to critically comment on the conflicting experimental results, highlighting the facts, the myths and the controversies. The goal is to indicate how to take advantage of chitosan versatility, to learn how to manage its variability and show how to properly tackle some unexpected undesirable features. In the sections of the paper various issues that relate chitosan properties to some basic features and to advanced solutions and applications are presented. The introduction outlines some historical pioneering works, where the chemistry of chitosan was originally explored. Thereafter, particular reference is made to analytical purity, characterization and chain modifications. The macromolecular characterization is mostly related to molecular weight and to degree of acetylation, but also refers to the conformational and rheological properties and solution stability. Then, the antimicrobial activity of chitosan in relation with its solubility is reviewed. A section is dedicated to the formulation of chitosan biomaterials, from gel to nanobeads, exploring their innovative application as active carrier nanoparticles. Finally, the toxicity issue of chitosan as a polymer and as a constructed nanomaterial is briefly commented in the conclusions.

Highlights: [•] Pectin was extracted from cocoa husks, a processing waste from the cocoa industry. [•] Citric acid is the most efficacious matrix for pectin extraction from cocoa husks. [•] Low methoxyl pectin from cocoa husks was obtained using water or hydrochloric acid. [Copyright &y& Elsevier]

Chitosan, copolymer of glucosamine and N-acetyl glucosamine is mainly derived from chitin, which is present in cell walls of crustaceans and some other microorganisms, such as fungi. Chitosan is emerging as an important biopolymer having a broad range of applications in different fields. On a commercial scale, chitosan is mainly obtained from crustacean shells rather than from the fungal sources. The methods used for extraction of chitosan are laden with many disadvantages. Alternative options of producing chitosan from fungal biomass exist, in fact with superior physico-chemical properties. Researchers around the globe are attempting to commercialize chitosan production and extraction from fungal sources. Chitosan extracted from fungal sources has the potential to completely replace crustacean-derived chitosan. In this context, the present review discusses the potential of fungal biomass resulting from various biotechnological industries or grown on negative/low cost agricultural and industrial wastes and their by-products as an inexpensive source of chitosan. Biologically derived fungal chitosan offers promising advantages over the chitosan obtained from crustacean shells with respect to different physico-chemical attributes. The different aspects of fungal chitosan extraction methods and various parameters having an effect on the yield of chitosan are discussed in detail. This review also deals with essential attributes of chitosan for high value-added applications in different fields. [ABSTRACT FROM AUTHOR]

Chitosan is a biopolymer of increasing significance, as well as a renewable and sustainable material. Its main molecular characteristics are molar mass and degree of acetylation (composition). Precise average degrees of acetylation were measured by quantitative H solution-state NMR spectroscopy. While number-average degrees of acetylation had already been determined by H NMR spectroscopy, weight-average degrees of acetylation are also determined and may be more relevant for some properties, such as mechanical properties. We report the first separation of chitosan according to its degree of acetylation using free solution capillary electrophoresis. Capillary electrophoresis separates chitosan in the 'critical conditions': the molar mass plays little role and the separation is by the degree of acetylation. It characterises the heterogeneity of chitosan samples in terms of composition (dispersity of the distribution of degrees of acetylation). This heterogeneity (broad distribution of degrees of acetylation) cannot be neglected contrary to a common assumption found in the literature. This fast and easy separation will allow establishing a structure-property relationships. [Figure not available: see fulltext.] [ABSTRACT FROM AUTHOR]

The mutant ATCN4 strain of Azotobacter vinelandii, which lacks the Na-NQR activity and results in an alginate overproduction (highly mucoid phenotype), was cultured in shake flasks in minimal and rich medium, and the chemical composition and rheological properties of the alginate were determined. Mutant ATCN4 exhibited a high efficiency for sucrose conversion to alginate and PHB accumulation, reaching yields that were 3.6- and 1.6-fold higher than those obtained from the wildtype cultures in minimal medium (Burk's sucrose, BS). The alginate produced by ATCN4 in the minimal medium had a high degree of acetylation (≥4 %) and a low G/M ratio (=2) with respect to the polymer synthesised in the rich medium (BS with yeast extract) (degree of acetylation = 0 % and G/M ratio of 4.5). The alginate produced in the minimal medium exhibited a pronounced pseudoplastic behaviour and a higher G* module in comparison to that observed in the alginate obtained in the cultures using a rich medium. The ATCN4 mutant culture in the minimal medium promoted the synthesis of a polymer of improved rheological quality in terms of its mechanical properties. These characteristics make this mutant a valuable source for producing alginates with improved or special properties. [ABSTRACT FROM AUTHOR]

The objectives of this study are to explore the effect of the ionic strength of aqueous acidic media and the intrinsic characteristics of chitosan used on the formation of different types of aggregation. Two different degrees of acetylation (DA) of β-chitosan are prepared from β-chitin by stepwise alkali deacetylation. These two chitosans have high chain stiffness and typical good solvent behavior in a 0.20 M acetic acid/0.10 M sodium acetate buffer solution. However, a trace amount of aggregates forms in the higher DA chitosan dilute solution. At ionic strength (I) ≥ 0.20 M, the association of compact chitosan forms and/or the amount of the aggregates increases with increasing I of the media. At I = 0.01 M, a considerable quantity of particle structure is observed in the low ionic strength solution and the electrodynamic coupling effect predominates on the mobile behavior of the extended and congested chitosan molecule in solution. [ABSTRACT FROM AUTHOR]

Abstract: Bacillus cereus is an endospore-forming bacterium able to cause food-associated illness. Different treatment processes are used in the food industry to reduce the number of spores and thereby the potential of foodborne disease. Chitosan is a polysaccharide with well-documented antibacterial activity towards vegetative cells. The activity against bacterial spores, spore germination and subsequent outgrowth and growth (the latter two events hereafter denoted (out)growth), however, is poorly documented. By using six different chitosans with defined macromolecular properties, we evaluated the effect of chitosan on Bacillus cereus spore germination and (out)growth using optical density assays and a dipicolinic acid release assay. (Out)growth was inhibited by chitosan, but germination was not. The action of chitosan was found to be concentration-dependent and also closely related to weight average molecular weight (Mw) and fraction of acetylation (FA) of the biopolymer. Chitosans of low acetylation (FA =0.01 or 0.16) inhibited (out)growth more effectively than higher acetylated chitosans (FA =0.48). For the FA =0.16 chitosans with medium (56.8kDa) and higher Mw (98.3kDa), a better (out)growth inhibition was observed compared to low Mw (10.6kDa) chitosan. The same trend was not evident with chitosans of 0.48 acetylation, where the difference in activity between the low (19.6kDa) and high Mw (163.0kDa) chitosans was only minor. In a spore test concentration corresponding to 102–103 CFU/ml (spore numbers relevant to food), less chitosan was needed to suppress (out)growth compared to higher spore numbers (equivalent to 108 CFU/ml), as expected. No major differences in chitosan susceptibility between three different strains of B. cereus were detected. Our results contribute to a better understanding of chitosan activity towards bacterial spore germination and (out)growth. [Copyright &y& Elsevier]

Abstract: Chitosans, polysaccharides obtained from the exoskeleton of crustaceans, have been shown to exert antibacterial activity in vitro and their use as a food preservative is of growing interest. However, beyond a consensus that chitosan appears to disrupt the bacterial cell membrane, published data are inconsistent on the chemical characteristics that confer the antibacterial activity of chitosan. While most authors agree that the net charge density of the polymer (reflected in the fraction of positively charged amino groups at the C-2 position of the glucosamine unit) is an important factor in antibacterial activity, conflicting data have been reported on the effect of molecular weight and on the susceptibility among different bacterial species to chitosan. Therefore, we prepared batches of water-soluble hydrochloride salts of chitosans with weight average molecular weights (Mw) of 2–224kDa and degree of acetylation of 0.16 and 0.48. Their antibacterial activity was evaluated using tube inhibition assays and membrane integrity assays (N-Phenyl-1-naphthylamine fluorescence and potassium release) against Bacillus cereus, Escherichia coli, Salmonella Typhimurium and three lipopolysaccharide mutants of E. coli and S. Typhimurium. Chitosans with lower degree of acetylation (FA =0.16) were more active than the more acetylated chitosans (FA =0.48). No trends in antibacterial action related to increasing or decreasing Mw were observed although one of the chitosans (Mw 28.4kDa, FA =0.16) was more active than the other chitosans, inhibiting growth and permeabilizing the membrane of all the test strains included. The test strains varied in their susceptibility to the different chitosans with wild type S. Typhimurium more resistant than the wild type E. coli. Salmonellae lipopolysaccharide mutants were more susceptible than the matched wild type strain. Our results show that the chitosan preparation details are critically important in identifying the antibacterial features that target different test organisms. [Copyright &y& Elsevier]

Abstract: Enzymatic defense responses and protection against Phytophthora nicotianae were studied in tobacco plants treated with chitosan of different molecular weights (MW) and degrees of acetylation (DA). The concentration and mode of chitosan application affected enzymatic induction in tobacco leaves. β-1,3-Glucanase (EC 3.2.1.6) activity required 10 times the polymer concentration relative to the oligochitosan mixture to induce the highest activity above control when treated by foliar spray, indicating the influence of molecular weight in this response. PAL (EC 4.3.1.5) and POD (EC 1.11.1.6) activities increased above control as a result of the influence of polymer degree of acetylation when treatments were applied by foliar spray. A higher DA favored PAL activity, whereas a lower DA induced higher POD activity. Using an in vitro bioassay, it was found that the three chitosan compounds caused a reduction of the infection index of P. nicotianae in tobacco plants that was dependent on elicitor concentrations. There was a significant relationship between the reduction of the infection index and an increase in PAL activity when chitosan was applied by foliar spray and by substrate drench to the plant rhizosphere. These results demonstrate the influence of chitosan physico–chemical properties in plant-induced resistance and the relevance of particular responses in plant protection against pathogens. [Copyright &y& Elsevier]

The dissolution behavior of chitin in a series of ionic liquids containing alkylimidazolium chloride, alkylimidazolium dimethyl phosphate, and 1-allyl-3-methyl-imidazolium acetate has been studied. Chitin with a low degree of acetylation and low molecular weight can be readily dissolved in ionic liquids, while those with a high degree of acetylation can also be dissolved in ionic liquids with various critical solution temperatures (CST) mainly depending on the structure of the chitin and the ionic liquids. The results indicated that the dissolution behavior of chitin in ionic liquids was affected by the degree of acetylation (DA), the crystallinity, and the molecular weights of chitin, as well as the nature of the anion of the ionic liquid. [ABSTRACT FROM AUTHOR]

The aim of this work is to investigate the gelation behavior of chitosans which has various degree of acetylation (DA) of amino group to ensure the preparation of the designed membrane structure. The DA was controlled by the amount of acetic anhydride added to the solution of chitosan/acidified water/methanol mixture. The gelation behavior was evaluated by the gelation time and the quantity of syneresis. The mechanical strength of acetylated chitosan gels was also measured. The optimum condition for the formation of the membrane, i.e. chitosan concentration, acetic acid/ chitosan mass ratio, methanol/water mass ratio, and amount of acetic anhydride for acetylation, was determined. This composition gives low contractility, shorter gelation time, and high mechanical strength to the membrane and the gel. Useful information not only for a preparation of a membrane but also for an immobilized carrier or a chemical reaction system was obtained in this work. [ABSTRACT FROM AUTHOR]

Abstract: In this study, a new source of insect chitin was proposed. Insect chitin was extracted from cicada sloughs by 1M HCl and 1M NaOH treatment for demineralization and deproteinization, respectively. The brown color of this chitin from cicada sloughs was removed using 6% sodium hypochlorite as an oxidizing agent. It was found that the insect chitin extracted from the cicada sloughs has a higher percent recovery than the chitin from rice-field crab shells. The chemical structure and physicochemical properties of α-chitin from cicada sloughs were characterized using elemental analysis (EA), attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR), proton nuclear magnetic resonance spectroscopy (1H NMR), solid-state 13C cross-polarization magic-angle-spinning nuclear magnetic resonance (CP/MAS) NMR spectroscopy, X-ray diffractometry (XRD), and thermogravimetry (TG). The degree of acetylation (DA) was determined by EA, 1H NMR, and 13C CP/MAS NMR techniques. The DA values of chitin from cicada sloughs were in the range of 97% to 102% depending on each technique. Furthermore, it was found that the DA increased with an increasing thermal property and crystallinity. [Copyright &y& Elsevier]

Abstract: An acetylesterase was purified from Aspergillus aculeatus on the basis of its ability to deacetylate homogalacturonan. A series of well-characterized substrates was prepared and used to investigate its specificity, including pectin with various degrees of methylation and acetylation, pectic domains with various structures, and oligomers. It was then compared to a rhamnogalacturonan acetylesterase previously isolated from the same fungus. Both enzymes were active towards various acetylated pectins, and were able to release acetyl groups from acetylated homogalacturonan, oligogalacturonates as well as rhamnogalacturonan at different extent. It was thus concluded that while the two enzymes differed in their efficiency, they are both pectin acetylesterases, able to act in the “smooth” as well as in the “hairy” regions of pectin. [Copyright &y& Elsevier]

Abstract: An economical and accurate determination of degree of acetylation (DA) for highly acetylated chitin has always been a challenge for researchers dealing with chitin and chitosan. A new protocol for the first derivative UV method using concentrated phosphoric acid as a solvent for highly acetylated chitin was developed in this study. The solvent was proposed based on thorough investigation of the effects of associated reactions including chain degradation, monomer dehydration, and oxazolinium ion formation. The reproducibility and performance of the new protocol was evaluated using commercial samples and the results showed the DA values of both chitin and chitosan could be determined accurately by a single analytical technique in less than 3h. [Copyright &y& Elsevier]

Chitosans with different degree of acetylation (DA, 10–50%) were synthesized by the acetylation reaction of deacetylated chitosan and acetic anhydride with different ratios. The porous beads (approx. 500 μm) fabricated from the acetylated chitosans were used to investigate the degradation behaviors of chitosans with different DA in vitro and in vivo. The in vitro degradation behavior of the acetylated chitosan beads was investigated in solutions of lysozyme and/or N-acetyl-β-D-glucosaminidase (NAGase), which are enzymes for chitosan present in the human body. It was observed that the degradation rate of acetylated chitosans can be controlled by adjusting the DA value: the degradation increased with increasing DA value of the acetylated chitosans. It seemed that NAGase plays an important role for the full degradation of chitosans in the body, even though NAGase itself can not initiate the degradation of chitosans. The in vitro degradation behavior of the chitosans in the mixture solution of lysozyme and NAGase was more similar to the in vivo degradation behavior than in the single lysozyme or NAGase solution. It may be owing to the sequential degradation reaction of chitosans in the mixture solution of lysozyme and NAGase (initial degradation by lysozyme to low-molecular-weight species or oligomers and the following degradation by NAGase to monomer forms). The in vivo degradation rate of acetylated chitosan beads was faster than the in vitro degradation rate. The acetylated chitosan porous beads with different DA value (and thus different degradation time) can be widely applicable as cell carriers for tissue-engineering applications. [ABSTRACT FROM AUTHOR]

Chitosan (Ch) is being actively investigated as a non-protein template for the growth of an increasing number of anchorage-dependent cells, including chondrocytes and bone cells. In the present work, Ch films with degrees of N-acetylation (DAs) in the range of 4 to 49% were evaluated with respect to the attachment, spreading and short-term proliferation of osteoblasts, using human osteoblastic MG-63 cells. The films were characterized in terms of surface morphology and surface charge by atomic force microscopy and streaming potential measurements, respectively. Cell attachment was assessed after 3 and 24 h of cell culture. After 24 h of incubation, cell attachment was found to be dependent on the DA, lower DAs favouring cell adhesion. With time, cell spreading and cytoskeleton organization were only attained for DAs ≤ 13%. Regarding cell proliferation, cells grown on films with the lowest DA (4%) revealed a higher specific growth rate as compared to those grown on films with higher DAs. Films with a DA of 49% failed to sustain cell proliferation. In addition, a longer lag-phase was observed on Ch, as compared to TCPS, in accordance to an observed delay of cell spreading. The present findings revealed that differences in the DA as small as 9% may be critical in terms of the osteoblast response to two-dimensional Ch-based matrices. [ABSTRACT FROM AUTHOR]