Extraction Of Chondroitin Sulfate Research Articles

Extraction of chondroitin sulfate and type II collagen from sturgeon (Acipenser gueldenstaedti) notochord and characterization of their hybrid fibrils

Chondroitin sulfate (CS) and undenatured type II collagen (Col II) are the two major, biological macromolecules of cartilage-related tissues. In this study, a new extraction. process was developed to obtain CS and Col II simultaneously. By this process, CS. and undenatured Col II were extracted from sturgeon notochord with the yields of 5.34, ± 0.74% and 45.25 ± 5.25%, respectively. The SEC-RI-MALLS result showed that the, average molecular weight of notochord CS was 38.4 kDa. FTIR NMR, and SAX-HPLC, results indicated the notochord CS was mainly composed of CS-A. The new extraction, process had no effect on the triple helical structure of Col II. To analyze the interaction, between the two macromolecules, the effect of CS on Col II fibril formation was, examined using turbidity assay and SEM observation. CS accelerated the completion, of Col II self-assembly and inhibited the lateral aggregation of fibrils. The results of this, study suggested that the sturgeon notochord is a valuable source of CS and Col II. The. new extraction method not only improves the utilization rate of sturgeon notochord, but,also reduces the waste of aquatic resources. CS and Col II derived from sturgeon, notochord have the potential for use in biomedical materials

Structural Characterization and Glycosaminoglycan Impurities Analysis of Chondroitin Sulfate from Chinese Sturgeon.

Chinese sturgeon was an endangered cartilaginous fish. The success of artificial breeding has promoted it to a food fish and it is now beginning to provide a new source of cartilage for the extraction of chondroitin sulfate (CS). However, the structural characteristics of sturgeon CS from different tissues remain to be determined in more detail. In this study, CSs from the head, backbone, and fin cartilage of Chinese sturgeon were individually purified and characterized for the first time. The molecular weights, disaccharide compositions, and oligosaccharide sulfation patterns of these CSs are significantly different. Fin CS (SFCS), rich in GlcUAα1-3GalNAc(4S), has the biggest molecular weight (26.5 kDa). In contrast, head CS (SHCS) has a molecular weight of 21.0 kDa and is rich in GlcUAα1-3GalNAc(6S). Most features of backbone CS (SBCS) are between the former two. Other glycosaminoglycan impurities in these three sturgeon-derived CSs were lower than those in other common commercial CSs. All three CSs have no effect on the activity of thrombin or Factor Xa in the presence of antithrombin III. Hence, Chinese sturgeon cartilage is a potential source for the preparation of CSs with different features for food and pharmaceutical applications.

Preparation and characterization of chondroitin sulfate from large hybrid sturgeon cartilage by hot-pressure and its effects on acceleration of wound healing

In this paper, a combination of hot-pressure, enzymatic hydrolysis and membrane separation process is used for efficiently and environmentally friendly extraction of chondroitin sulfate (CS) from large hybrid sturgeon cartilage, namely, HPCS. The recovery and yield of CS were 93.68% and 36.47% under the optimized conditions. Fourier transform infrared (FT-IR) spectroscopy, nuclear magnetic resonance (NMR) spectroscopy and high-performance liquid chromatography (HPLC) indicated that the HPCS was composed of monosulfated disaccharides in position 6 and 4 of the N-acetyl-D-galactosamine (58.38% and 27.34%, respectively) and nonsulfated disaccharide (14.29%), which was similar to the composition of CS extracted by dilute alkali-enzymatic hydrolysis-chemical precipitation from large hybrid sturgeon cartilage (SCS). The wound healing results indicated that HPCS could promote cell migration and proliferation, alleviate inflammation and facilitate angiogenesis, which results in its excellent wound treatment activity. These results provide theoretical and practical significance for the production and application of chondroitin sulfate.

Optimization of Enzyme Assisted Extraction of Chondroitin Sulfate from Bohadschia argus by Response Surface Methodology

Chondroitin sulfate is currently one of the bioactive compounds obtained from different natural materials with high benefit in human medical treatment and pharmaceutical. However, the natural concentration of chondroitin sulfate in high cost raw material is low and this situation makes the commercialization become difficult. This study focused on production of chondroitin sulfate isolated from Bohadschia argus using enzyme-assisted extraction methodology. Optimization experiments were conducted based on Response Surface Methodology (RSM) using Box–Behnken design (BBD). Three important extraction parameters, including enzyme concentration (X1: 0.05-0.5 %), Time (X2: 0.25–3 h), and temperature (X3: 55–65 °C) were varied to obtain maximum chondroitin sulfate yields. The RSM optimized model obtained from statistical analysis presented the high correlation coefficiency (R2) at 0.7508, advocating the significance of the model. The optimum extraction conditions were selected as enzyme concentration of 0.48% with extraction time of 1.01 h. at an extraction temperature of 56.53°C. Under these conditions, the experimental yield of chondroitin sulfate was 415.59 mg/100g dry, which is in good agreement with the value predicted by the model.

Extraction of hyaluronic acid and chondroitin sulfate from marine biomass for their application in the treatment of the dry eye disease

AbstractPurposeTear film instability, hyperosmolarity and neurosensory abnormality are the key etiological contributors to the pathophysiological process of the Dry Eye Disease (DED).1 Hyaluronic acid (HA) and Chondroitin sulfate (CS) have shown improvements in the corneal epithelial barrier in patients with DED.2 Moreover they are used in pharmaceutical applications due to their biocompatibility, viscoelasticity, lubricity and immunostimulatory effects. In this study, they are extracted from marine wastes and will be evaluated as possible DED treatment using natural, green and novel Deep Eutectic Solvents (DES) to replace time‐consuming and expensive conventional methods and maintaining the quality and purity of the isolated HA and CS. Natural compounds such as menthol and trehalose, are used in DES preparation and have shown to be effective in DED treatment.3,4MethodsHA and CS are isolated from marine wastes using conventional methods and DES as low‐cost alternative technique. DES are prepared by heating and mixing the components at certain ratios. They are mixed with the marine sources at specific extraction conditions. The extracts are then enzymatically hydrolyzed and analyzed by HPLC and capillary electrophoresis to quantify HA and CS.ResultsHA and CS are extracted from mussels, codfish bones and furtherly tuna eyeballs and shark fins using DES. Several DES were tested such as menthol:borneol, menthol:camphor, trehalose:glycerol, and significant amounts of HA and CS were extracted with high specificity. For example, 37.92 µg/gdry codfish bones CS disaccharide (ΔHexAβ1‐3GalNAc4S) were extracted using menthol:borneol.ConclusionDES are effective compounds for the extraction of valuable biopolymers HA and CS. The isolated HA and CS, as well as their therapeutic mixture with DES will be studied in vitro on conjunctival and corneal epithelial human cell lines to assess their toxicity, biocompatibility, muco‐adhesion and anti‐oxidant effects for the treatment of the DED.References Craig, Jennifer P., et al. “TFOS DEWS II report executive summary.” The Ocular Surface 15.4 (2017): 802‐812. Limberg, Michael B., et al. “Topical application of hyaluronic acid and chondroitin sulfate in the treatment of dry eyes.” American journal of ophthalmology 103.2 (1987): 194‐197. Arita, Reiko, et al. “Effects of a warm compress containing menthol on the tear film in healthy subjects and dry eye patients.” Scientific reports 7 (2017): 45848. Fondi, Klemens, et al. “Effect of Hyaluronic Acid/Trehalose in Two Different Formulations on Signs and Symptoms in Patients with Moderate to Severe Dry Eye Disease.” Journal of ophthalmology 2018 (2018).

Characteristics of Chondroitin Sulfate Extracted of Tilapia (Oreochromis niloticus) Processing

In the present work, an alternative value aggregation to the residues from the industrialization of tilapia is suggested by extraction of chondroitin sulfate from the bones of the spine. Resulting materials were chemically characterized by thermogravimetry (TGA/DSC), infrared spectroscopy (FTIR) and scanning electron microscopy (SEM). Through the characterization of CS, it was possible to conclude that the methodology used to extract it generated a product with high thermal stability and purity.

Optimisation extraction of chondroitin sulfate from fish bone by high intensity pulsed electric fields

High intensity pulsed electric fields (PEF) was used to extract chondroitin sulphate (CS) from fish bone. Results show that PEF extraction speed is much faster, and the content of CS is much higher compared with traditional methods. Variation of PEF parameters and the content of CS were determined by single factor experiments. The processing conditions were optimised by quadratic general rotary unitised design experiments. The maximum yield of 6.92g/L was achieved under the following conditions: material–liquid ratio of 1:15g/mL, electric field intensity of 16.88kV/cm, pulse number of 9, and NaOH concentration of 3.24%. The purity of CS was analysed by agarose gel electrophoresis. CS purity was high, and the extract did not contain any other glycosaminoglycans. PEF can be widely used to extract CS with non-thermal performance, high speed, and low pollution.

Extraction of Chondroitin Sulfate from Tilapia Byproducts with Ultrasonic-Microwave Synergistic

The optimum conditions for the extraction and separation of chondroitin sulfate from tilapia (Oreochromis niloticus) byproducts were studied in the paper. The chondroitin sulfate was extracted by the combination of ultrasonic and microwave. On the basis of the single factor tests, the response surface methodology was taken to study the optimum process of extraction. Results show that the best conditions for the extraction and separation of chondroitin sulfate from tilapia byproducts are: the ultrasonic power is 50W, the microwave power is 98W, the extraction time is 120s, the ratio of solid to liquid is 1:50 (g/mL), and under these conditions, the extraction rate of chondroitin sulfate is 2.513%.

Extraction, purification and characterisation of chondroitin sulfate in Chinese sturgeon cartilage

Chinese sturgeon (Acipenser sinensis), a rare species, is an important fishery resource in China. To save this species from overfishing, damming and pollution, artificial propagation has been developed rapidly in recent years. However, the cartilage from Chinese sturgeon, which contains an abundance of chondroitin sulfate (CS), is currently discarded as solid waste after consumption of the fish. The aim of this study was to investigate the extraction, purification and characterisation of CS from Chinese sturgeon cartilage. The optimal extraction parameters were a ratio of aqueous NaOH to cartilage powder of 9.2, a concentration of aqueous NaOH of 4.4% and an extraction time of 3.9 h. Under these optimal conditions the yield of crude CS from Chinese sturgeon cartilage was 26.51%, in agreement with the model prediction (26.54%). Purification by DEAE-52 cellulose and Sephadex G-100 column chromatography yielded a single fraction, CS-11. Its characterisation indicated that CS-11 was consistent with the polysaccharide backbone structure -4GlcAβ-3GalNβ- and was present in the form of chondroitin-4-sulfate and chondroitin-6-sulfate. The results of this study provide a basis for promoting the utilisation of Chinese sturgeon resources and are significant for the development and utilisation of CS from Chinese sturgeon cartilage in the food industry.

Evaluation of increasing peptone blend on nursery pig performance from 15 to 40 lb (2013)

A total of 270 pigs (PIC 327 × 1050, initially 15.7 lb BW) were used in a 28-d trial to evaluate the effects of increasing levels of a new peptone blend by-product on nursery pig growth performance. The product is the result of the pharmaceutical extraction of chondroitin sulfate from bovine cartilage and processing to form the peptone blend, which was mixed with soybean hulls and drum-dried. Pigs were weaned at 21 d of age and were fed a common pelleted diet for 5 d prior to the start of the experiment. Each treatment had 8 replicate pens and 6 or 7 pigs per pen. The 5 experimental treatments were: (1) a diet with 1% blood meal and 2% select menhaden fish meal (positive control), (2) a diet with no added specialty protein source (negative control), (3) a diet containing 4% peptone blend, (4) a diet containing 8% peptone blend, or (5) a diet containing 12% peptone blend. Experimental diets were fed for 14 d, then a common Phase 2 diet was fed for an additional 14 d to determine the residual treatment effects on growth performance.

Chondroitin sulfate extraction from broiler chicken cartilage by tissue autolysis

Chondroitin sulfate (CS) is an acidic polysaccharide comprised of repeating disaccharide units of N-acetylgalactosamine and glucuronic acid. It is known as a dietary supplement for joint articular cartilage care. CS is commonly extracted from bovine and shark cartilages by digestion of tissues with exogenous proteinase. This study was undertaken to extract CS from broiler chicken cartilage by tissue autolysis without using exogenous proteinase. Most (>90%) of tissue uronic acid was extracted by incubation with 0.5 mol/L sodium acetate at pH 4.5 and 37 °C overnight followed by tissue reextraction. There was no difference (P > 0.05) in the proportion of extractable uronic acid among cartilage tissues from different anatomical locations. The final product obtained after anion exchange chromatography was a large CS-peptide with glucuronosyl-N-acetylgalactosamine-4-sulfate as a predominant disaccharide. Keratan sulfate was detected by hexosamine analysis, susceptibility to keratanase, and immunodiffusion with anti-keratan sulfate monoclonal IgM. The CS-peptide also contained serine as a predominant amino acid accounting for approximately one third of total amino acid. It was concluded that tissue autolysis is a simple efficient procedure that can be scaled up for extraction of broiler chicken CS.

Solvent-free mechanochemical extraction of chondroitin sulfate from shark cartilage

A solvent-free mechanochemical extraction (SFMCE) technology was applied to the extraction of chondroitin sulfate (CS) from shark cartilage. This process was carried out via mechanical pretreatment (MPT) and mechanochemical treatment (MCT) in AGO-2 high-intensity planetary activator at room temperature under solvent-free condition. The yield of CS was increased from 8.50 ± 0.16% to 9.33 ± 0.14% and the purity of product was improved from 86.4 ± 0.88% to 95.1 ± 0.65%. The effectiveness of the MCT was confirmed by HPLC–ELSD analysis. The energy cost was decreased obviously due to the decrease of extraction time from 3 h to 3 min. SFMCE method was proved to be a high-efficient and energy-saving technique.

Determination of chondroitin sulfate from different sources of cartilage

Cartilage is produced as a by-product from slaughter house and fishery industries in Thailand. The enzymatic extraction of chondroitin sulfate followed the method of Nakano et al. [T. Nakano, H.H. Sunwoo, X. Li, M.A. Price, S.S. Jeong, Study of sulfated glycosaminoglycans from porcine skeletal muscle epimysium including analysis of iduronosyl and glucoronosyl residues in galactosaminoglycan fractions, J. Agric. Food Chem. 44 (1996) 1424–1434] and the sulfate glycosaminoglycans (GAGs) assay [W.R. Farndale, D.J. Buttle, A.J. Barrett, Improved quantitation and discrimination of sulphated glycosaminoglycans by use of dimethylmethylene blue, Biochim. Biophys. Acta 883 (1986) 173–177] was employed to determine the content of chondroitin sulfate in cartilage of shark fin, ray, crocodile and chicken keel. Identification of types of chondroitin sulfate through FTIR spectroscopy KBr pellet technique was carried out. The results indicated that chicken keel, crocodile hyoid and sternum cartilage are the most promising potential sources of chondroitin sulfate. The value ranges from 11.55 to 14.84 g/100 g of dried cartilage, calculated as chondroitin-4-sulfate. Identification of dried chondroitin sulfate extracts from investigated cartilages show the existence of both chondroitin-4-sulfate and chondroitin-6-sulfate.

Enzymatic extraction of chondroitin sulfate from skate cartilage and concentration-desalting by ultrafiltration

Skate cartilage is a fishery by-product, which contains chondroitin sulfate (CS), a glycosaminoglycan well known for its chondroprotective effect. Here described is a low-cost two-step process producing CS in non-denaturing conditions, consisting of an enzymatic extraction followed by tangential filtration to concentrate and purify CS. The performances of UF and MF membranes were compared in terms of flux and selectivity. The 0.1 μm-pore size membrane appeared to be the most efficient to separate CS from the other compounds.

軟骨からのコンドロイチン硫酸の抽出におけるアルカリ処理に関する研究-I

The influence of alkali treatment on the behavior of chondroitin sulfate (CS), kera-tosulf ate (KS) and protein in Dowex 50 W treatment and ethanol precipitation, which are the main processes for the extraction of CS, were studied and the following results were obtained. 1) Some differences in elution patterns of CS, KS and protein from a column of Dowex 50 W-X l(H+) were perceived between the liquefied whale nasal cartilage solu-tion and alkali-treated ones. CS was scarcely adsorbed on the Dowex 50 W column, but some amounts of KS and a greater part of protein were adsorbed. 2) The ethanol concentration needed for the precipitation of CS in the alkali-treated whale nasal cartilage mucopolysaccharide mixture was almost the same as that in the untreated preparation, while the ethanol concentration needed for the precipitation of KS was higher. It may be concluded that CS is easily separated from a greater part of KS by precipitating with ethanol after alkali treatment.