Determination of molecular mass values of chondroitin sulfates by fluorophore-assisted carbohydrate electrophoresis (FACE)

Buzzega, Dania; Maccari, Francesca; Volpi, Nicola

doi:10.1016/j.jpba.2009.10.015

Cited by 13 publications

(10 citation statements)

References 9 publications

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“…FTIR spectra (Supporting Information Figure 6) of HAP revealed the presence of expected bonds in the crystal. Strong modes for stretching (1035 cm –1 ) and bending (564, 604 cm –1 ) indicated the presence of PO 4 . − Characteristic stretching (3572 cm –1 ) of the −OH group was also observed. , Additionally, some bands indicating the weak presence of adsorbed water (1635 and 3000–3700 cm –1 ) were observed. , …”

Section: Resultsmentioning

confidence: 93%

“…These GAGs exhibit desirable physicochemical properties as scaffolds in tissue engineering. ,− ,, HA is a linear, high-molar-mass polysaccharide composed of alternating (1 → 4)-β-linked d -glucuronic acid and (1 → 3)-β-linked N -acetyl- d -glucosamine residues. CS differs only in the N -acetyl- d -glucosamine residue, which is sulfonated at either the 4 or 6 carbon site. ,,, Because of repeating carboxyl or sulfonate moieties along the backbone, HA and CS occur as anionic macromolecules in aqueous environments at extracellular pH. ,,, As a result, these highly charged macromolecules possess desirable shear-thinning and viscoelastic properties − ,, and have primarily been used as inert carrier fluids in bone tissue engineering applications . Emerging efforts, however, aim to elicit “bridging” connections within colloidal gels by using particles to act as anchoring points between multivalent, adsorbing polymers. ,,− ,− ,,, …”

Section: Introductionmentioning

confidence: 99%

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Mapping Glycosaminoglycan–Hydroxyapatite Colloidal Gels as Potential Tissue Defect Fillers

et al. 2014

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Malleable biomaterials such as Herschel–Bulkley (H–B) fluids possess shear responsive rheological properties and are capable of self-assembly and viscoelastic recovery following mechanical disruption (e.g., surgical placement via injection or spreading). This study demonstrated that the addition of moderate molecular weight glycosaminoglycans (GAGs) such as chondroitin sulfate (CS) (Mw = 15–30 kDa) and hyaluronic acid (HA) (Mw = 20–41 kDa) can be used to modify several rheological properties including consistency index (K), flow-behavior index (n), and yield stress (τy) of submicrometer hydroxyapatite (HAP) (Davg ≤ 200 nm) colloidal gels. GAG–HAP colloidal mixtures exhibited substantial polymer–particle synergism, likely due to “bridging” flocculation, which led to a synergistic increase in consistency index (KGAG-HAP ≥ KGAG + KHAP) without compromising shear-thinning behavior (n < 1) of the gel. In addition, GAG–HAP colloids containing high concentrations of HAP (60–80% w/v) exhibited substantial yield stress (τy ≥ 100 Pa) and viscoelastic recovery properties (G′recovery ≥ 64%). While rheological differences were observed between CS–HAP and HA–HAP colloidal gels, both CS and HA represent feasible options for future studies involving bone defect filling. Overall, this study identified mixture regions where rheological properties in CS–HAP and HA–HAP colloidal gels aligned with desired properties to facilitate surgical placement in non-load-bearing tissue-filling applications such as calvarial defects.

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Section: Resultsmentioning

confidence: 93%

Section: Introductionmentioning

confidence: 99%

Mapping Glycosaminoglycan–Hydroxyapatite Colloidal Gels as Potential Tissue Defect Fillers

et al. 2014

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“…The molecular mass of CS/DS obtained from Atlantic bluefin tuna skins (SGAT) was determined by PAGE according to Edens et al 29 15 μg of the purified CS/DS determined by uronic acid assay were layered on the gel. The related calibration curve was constructed using oligosaccharide standards of known molecular mass prepared from CS Buzzega et al 30 After a run of 40 min at 100 V, the gel was stained with toluidine blue (0.1% in 1% acetic acid) for 30 min, followed by destaining in 1% acetic acid. Molecular mass evaluation was performed by densitometric acquisition of the bands and comparison of their migration times on the calibration curve constructed by plotting the retention times of the standards against the logarithms of their molecular mass values.…”

Section: Methodsmentioning

confidence: 99%

Purification and structural elucidation of chondroitin sulfate/dermatan sulfate from Atlantic bluefin tuna (Thunnus thynnus) skins and their anticoagulant and ACE inhibitory activities

et al. 2018

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Chondroitin sulfate/dermatan sulfate (CS/DS) was extracted from Atlantic bluefin tuna (Thunnus thynnus) skin (SGAT) and was purified and characterized. SGAT was characterized by acetate cellulose electrophoresis, FTIR spectroscopy, 13 C NMR spectroscopy and SAX-HPLC. According to the results obtained for specific chondroitinases (ABC and AC) and the SAX-HPLC separation of generated unsaturated repeating disaccharides, the polymer was found to contain a disaccharide monosulfated in positions 6 and 4 of GalNAc and disulfated disaccharides in different percentages. These results were confirmed by 13 C NMR experiments. The average molecular mass was 24.07 kDa, as determined by PAGE analysis. SGAT was evaluated for its in vitro anticoagulant activity via activated partial thromboplastin time, thrombin time and prothrombin time tests. The polymer showed strong inhibitory activity against angiotensin I-converting enzyme (IC 50 ¼ 0.25 mg mL À1 ). Overall, the results suggest that this newly extracted CS/DS can be useful for pharmacological applications. Fig. 3 (A) Anticoagulant activity of the purified CS/DS from Atlantic bluefin tuna skin (SGAT) at different concentrations evaluated by measurement of the activated partial thromboplastin time (aPTT). (B) Anticoagulant activity of SGAT at different concentrations evaluated by measurement of the thrombin time (TT). (C) Anticoagulant activity of SGAT at different concentrations evaluated by measurement of the prothrombin time (PT). All results are expressed as mean AE SD (n ¼ 3) and all measurements were performed in triplicate. 37972 | RSC Adv., 2018, 8, 37965-37975 This journal is

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“…According to this, a combination of specific analytical methods to define the final quality, origin and properties of the product is required. Such methods include electrophoresis, HPSEC, selective enzymatic treatment and further separation of the constituent disaccharides/oligosaccharides using sophisticated techniques including capillary electrophoresis [31,52,53], HPLC [5,6,54,55], FACE (Fluorophore-Assisted Carbohydrate Electrophoresis) [42,56] and mass spectrometry [57]. CS profile is achieved using well-established multi-analytical laboratory techniques, particularly electrophoresis, HPSEC and eHPLC (Figure 8) [5,6] due to their high throughput, sensitivity and reproducibility.…”

Section: Analytical Quality Controlsmentioning

confidence: 99%

Chondroitin Sulfate Safety and Quality

Volpi

2019

Molecules

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The industrial production of chondroitin sulfate (CS) uses animal tissue sources as raw material derived from different terrestrial or marine species of animals. CS possesses a heterogeneous structure and physical-chemical profile in different species and tissues, responsible for the various and more specialized functions of these macromolecules. Moreover, mixes of different animal tissues and sources are possible, producing a CS final product having varied characteristics and not well identified profile, influencing oral absorption and activity. Finally, different extraction and purification processes may introduce further modifications of the CS structural characteristics and properties and may lead to extracts having a variable grade of purity, limited biological effects, presence of contaminants causing problems of safety and reproducibility along with not surely identified origin. These aspects pose a serious problem for the final consumers of the pharmaceutical or nutraceutical products mainly related to the traceability of CS and to the declaration of the real origin of the active ingredient and its content. In this review, specific, sensitive and validated analytical quality controls such as electrophoresis, eHPLC (enzymatic HPLC) and HPSEC (high-performance size-exclusion chromatography) able to assure CS quality and origin are illustrated and discussed.

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Determination of molecular mass values of chondroitin sulfates by fluorophore-assisted carbohydrate electrophoresis (FACE)

Cited by 13 publications

References 9 publications

Mapping Glycosaminoglycan–Hydroxyapatite Colloidal Gels as Potential Tissue Defect Fillers

Mapping Glycosaminoglycan–Hydroxyapatite Colloidal Gels as Potential Tissue Defect Fillers

Purification and structural elucidation of chondroitin sulfate/dermatan sulfate from Atlantic bluefin tuna (Thunnus thynnus) skins and their anticoagulant and ACE inhibitory activities

Chondroitin Sulfate Safety and Quality

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