Validated high-performance anion-exchange chromatography with pulsed amperometric detection method for the determination of residual keratan sulfate and other glucosamine impurities in sodium chondroitin sulfate

Bottelli, Susanna; Grillo, Gianluca; Barindelli, Edoardo; Nencioni, Alessandro; Maria, Alessandro Di; Fossati, Tiziano

doi:10.1016/j.chroma.2017.04.045

Cited by 9 publications

(3 citation statements)

References 5 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Determination of the CS and GlcN contents of the sample supernatants and of their eventual KS contamination was performed by high-performance anion-exchange chromatography (HPAE-PAD) analysis on the basis of the monosaccharide composition, according to a previously reported method [23]. The CS content was determined after a hydrolytic procedure on the supernatants as previously described [23]; the GlcN monosaccharide content was determined after ultrafiltering the sample supernatants on 3-kDa membranes, according to a previously reported protocol [23, 26] and analyzing the permeate fractions. The retentate fraction that contained the CS and the eventual KS was hydrolyzed and analyzed as well.…”

Section: Methodsmentioning

confidence: 99%

“…Recent papers have also demonstrated that keratan sulfate (KS), which is another component of animal articular cartilaginous tissue, may be extracted along with the CS, thus contaminating it [23]. Because of the structural similarities, and the superimposable size of CS and KS biopolymers, the selective removal of this macromolecule is challenging [24–26]. It is of great scientific interest for the assessment of analytical strategies to fully characterize food supplement purity, with the specific target of finding a correlation between structural characteristic and biological activity using in vitro models [27].…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Comparative Analyses of Pharmaceuticals or Food Supplements Containing Chondroitin Sulfate: Are Their Bioactivities Equivalent?

et al. 2019

View full text Add to dashboard Cite

IntroductionOral supplementation of chondroitin sulfate (CS) and glucosamine (GlcN), symptomatic slow-acting molecules, is recommended by European Society for Clinical and Economic Aspects of Osteoporosis and Osteoarthritis and Musculoskeletal Diseases (ESCEO) and other European Union (EU) guidelines for the restoration of the articular cartilage surface in patients affected by osteoarthritis (OA). They are commercialized as pharmaceutical grade products and as food supplements in combination with plant extracts hyaluronic acid, methylsulfonylmethane, and other components. Food supplements do not need to undergo the strict regulatory controls of pharmaceutical grade products; thus, composition and contaminants that could be present may not be evidenced before commercialization and these uncertainties may give rise to concerns about the bioactivity of these formulations.MethodsIn this paper 10 different food supplements (FS) from diverse European countries were analyzed in comparison with two pharmaceutical grade products (Ph) using updated analytical approaches and biochemical cell-based assays. The purity, the titer, and the origin of CS in Ph and FS samples were initially assessed in order to successively compare the biological function. Both food supplements and pharmaceutical formulations were tested in vitro, using the same final CS concentration, on primary chondrocytes and synoviocytes in terms of (i) cell viability, (ii) activation of the NF-κB-mediated inflammation pathway, (iii) cartilage oligomeric matrix protein (COMP-2), IL-6, and IL-8 production.ResultsAll the FS presented a certain insoluble fraction; the CS and the GlcN contents were lower than the declared ones in 9/10 and 8/10 samples, respectively. All FS contained keratan sulfate (KS) at up to 50% of the total glycosaminoglycan amount declared on the label. Primary cells treated with the samples diluted to present the same CS concentration in the medium showed cytotoxicity in 7/10 FS while Ph preserved viability and reduced NF-κB, COMP-2, and secreted inflammatory cytokines.ConclusionAmong all samples tested, the pharmaceutical grade products demonstrated effective modulation of biomarkers counteracting the inflammation status and improving viability and the physiological condition of OA human primary chondrocyte and synoviocyte cells. In contrast to that, most FS were cytotoxic at the tested concentrations, and only 3/10 of them showed similarities to Ph sample behavior in vitro.FundingThis work was partially supported by PON01_1226 NUTRAFAST, MIUR Ministero dell’Università e della Ricerca Scientifica. Bioteknet financed two short-term grants for graduate technicians. The journal’s Rapid Service and Open Access fees were funded by IBSA CH.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Comparative Analyses of Pharmaceuticals or Food Supplements Containing Chondroitin Sulfate: Are Their Bioactivities Equivalent?

et al. 2019

View full text Add to dashboard Cite

show abstract

“…Pesek et al (2016) have isolated and determined glucosamine from shrimp shells on silica hydride-based stationary phase (aqueous normal-phase chromatography). Also ion-exchange stationary phase with pulsed amperometric detection has been reported (Cheng & Kaplan, 2003;Bottelli et al, 2017). For a direct measurement of glucosamine, special electrochemical sensors have also been developed (Suea-Ngam, Rattanarat, Wongravee, Chailapakul, & Srisa-Art, 2016;Kamyabi & Hajari, 2017).…”

Section: Various Methods For Determination Of Glucosaminementioning

confidence: 99%

Determination of glucosamine and monitoring of its mutarotation by hydrophilic interaction liquid chromatography with evaporative light scattering detector

Pazourek

2018

Biomedical Chromatography

View full text Add to dashboard Cite

Saccharides and their derivatives are typical polar analytes without a suitable UV-chromophore that are nowadays analyzed by HPLC (high-performance liquid chromatography) under HILIC (hydrophilic interaction liquid chromatography) mode. Usually an evaporative light scattering detector (ELSD) is utilized which, however, gives a nonlinear response. A procedure to overcome the problem of mutarotating (time-varying) analytes recorded with such a nonlinear response detector is described. The procedure was applied for determination of glucosamine in two commercially available pharmaceutical formulations containing the common inorganic ions that the detector gives a response to. Under optimized conditions, both the anomers of glucosamine were separated and could be determined separately. Owing to the short retention time of the analyte (a run time <4 min) and relatively slow kinetics of the anomeric conversion (equilibration time 2.5 h), mutarotation could be monitored and corresponding rate constants calculated.

show abstract

High-pH Anion-Exchange Chromatography (HPAEC) and Pulsed Amperometric Detection (PAD) for Carbohydrate Analysis

Hardy¹,

Rohrer

2021

Comprehensive Glycoscience

View full text Add to dashboard Cite

Validated high-performance anion-exchange chromatography with pulsed amperometric detection method for the determination of residual keratan sulfate and other glucosamine impurities in sodium chondroitin sulfate

Cited by 9 publications

References 5 publications

Comparative Analyses of Pharmaceuticals or Food Supplements Containing Chondroitin Sulfate: Are Their Bioactivities Equivalent?

Comparative Analyses of Pharmaceuticals or Food Supplements Containing Chondroitin Sulfate: Are Their Bioactivities Equivalent?

Determination of glucosamine and monitoring of its mutarotation by hydrophilic interaction liquid chromatography with evaporative light scattering detector

High-pH Anion-Exchange Chromatography (HPAEC) and Pulsed Amperometric Detection (PAD) for Carbohydrate Analysis

Contact Info

Product

Resources

About