Salt gradient chromatographic separation of chondroitin sulfate disaccharides

Tóth, Gábor; Vékey, Károly; Sugár, Simon; Kovalszky, Ilona; Drahos, László; Turiák, Lilla

doi:10.1016/j.chroma.2020.460979

Cited by 12 publications

(17 citation statements)

References 23 publications

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“…A Waters® nanoAcquity UPLC system (Waters, Milford, MA, USA) was coupled to a Waters® QTOF Premier™ Mass Spectrometer (Waters, Milford, MA, USA) via ESI source. The analysis was performed with a method previously reported [ 18 ]. Briefly, a 250 μm × 10.5 cm capillary column packed with GlycanPac AXH-1 resin (Unicam Plc., Debrecen, Hungary) was used at a temperature of 45 °C with a flow rate of 8 μL/min.…”

Section: Methodsmentioning

confidence: 99%

“…Various chromatographic methods (with UV or MS detection) have been reported to analyze sulfation patterns of CS disaccharides [ 10 , 11 ]. Some examples are the following: reversed-phase chromatography with derivatization or ion-pairing [ 12 , 13 ], size exclusion (SEC) [ 14 ], graphitized carbon [ 15 ], HILIC [ 16 , 17 ], or HILIC-WAX [ 18 , 19 ] chromatography. The most common injection solvents for the abovementioned methods are water, acetonitrile, methanol, and their mixtures with or without buffer salts, in order to match the initial conditions of the chromatographic run.…”

Section: Introductionmentioning

confidence: 99%

“…Volpi et al [ 20 ] investigated the stability of CS chains under acidic, neutral, and basic conditions, and Zaia et al [ 21 ] gave information on 4- O and 6- O sulfated CS disaccharides in 30:70 v/v% methanol:water injection solvent. Our group has previously shown the potential instability of CS disaccharides during storage at 4 °C in the injection solvent 75:25 v/v% acetonitrile:water (10 mM ammonium formate, pH = 4.4) used for their HILIC-WAX chromatographic method [ 18 ].…”

Section: Introductionmentioning

confidence: 99%

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Stability and recovery issues concerning chondroitin sulfate disaccharide analysis

et al. 2021

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Chondroitin sulfate (CS) is a widely studied class of glycosaminoglycans, responsible for diverse biological functions. Structural analysis of CS is generally based on disaccharide analysis. Sample preparation is a key analytical issue in this case. However, a detailed study on the stability and recovery of CS-derived species has been lacking so far. We have found that for solvent exchange, in general, vacuum evaporation (SpeedVac) is much preferable than lyophilization. Moreover, in the case of aqueous solutions, higher recovery was experienced than in solutions with high organic solvent content. Storage of the resulting disaccharide mixture in typical HPLC injection solvents is also critical; decomposition starts after 12 h at 4 °C; therefore, the mixtures should not be kept in the sample tray of an automatic injector for a long time. The study, therefore, lays down suggestions on proper sample preparation and measurement conditions for biologically derived chondroitin sulfate species.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Stability and recovery issues concerning chondroitin sulfate disaccharide analysis

et al. 2021

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“…CS is structurally based on N-acetylgalactosamine (GalNAc) as the neutral sugar component (MENDE et al 2016). CS is a class of long linear polysaccharides, composed of alternating saccharide units of GalNAc and glucuronic acid (GlcA) (TÓTH et al 2020).…”

Section: Chondroitin Sulfatementioning

confidence: 99%

“…Due to their large size, the verification of intact HS chains is practically impossible by instrumental analytical tools. Structural characterization of the average sulfation pattern is usually performed after enzymatic hydrolysis of the polymeric chain into the constituent disaccharide units (TÓTH et al, 2020). There are three enzymes with differing specificity for HS and Hp.…”

Section: Heparan Sulfatementioning

confidence: 99%

Properties, bioactive potential and extraction processes of glycosaminoglycans: an overview

et al. 2021

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Glycosaminoglycans (GAGs) are long-chain polysaccharides that are divided into sulphates and non-sulphates, these being chondroitin sulphate, heparan sulphate, dermatan sulphate, heparin sulphate and the only non-sulphate in the group is hyaluronic acid. GAGs are obtained from animal tissue and by an expensive low-yield extraction process; however, they are highly commercially valued polysaccharides and exploited in the biomedical market. Their disaccharidic composition, chain length and sulfation pattern present great variability depending on the species and extraction factors. GAGs possess immunomodulatory, antioxidant, antiviral, anti-inflammatory, neuroprotective, antiproliferative and anticoagulant properties, functioning as therapeutic agents modulating an array of biological processes. This report presents the general aspects of each GAG, source and extraction process, in addition to the characteristics that give them the most varied therapeutic properties and pharmacological applications.

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Investigation of sample handling steps for accurate heparan sulphate disaccharide analysis using HPLC–MS

Pál,

Tóth,

Turiák

2024

Electrophoresis

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Heparan sulphates (HSs), a specific class of glycosaminoglycans (GAGs), are important participants of cellular signalling. Analytical characterization of GAGs requires a complex sample preparation workflow. Although a detailed stability and recovery study is available for the chondroitin sulphate GAG class, the literature concerning HS is incomplete in this regard. Therefore, our aim was to systematically investigate various parameters that could potentially influence the stability and recovery of HS samples when performing disaccharide analysis using high‐performance liquid chromatography–mass spectrometry. First, effects concerning vacuum evaporation and freezing were investigated. Next, the storage stability of the HS disaccharides was analysed under several conditions such as temperature, pH, digestion buffers, injection solvents and storage vessels. We have identified several critical parameters influencing the stability and recovery of HS disaccharides. We concluded that major sample loss is expected when Tris–HCl is used as digestion buffer, followed by vacuum evaporation at elevated temperatures, or samples are stored under alkaline conditions. Following the practical considerations of this paper can contribute to increasing the reliability of future analytical measurements.

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Salt gradient chromatographic separation of chondroitin sulfate disaccharides

Cited by 12 publications

References 23 publications

Stability and recovery issues concerning chondroitin sulfate disaccharide analysis

Stability and recovery issues concerning chondroitin sulfate disaccharide analysis

Properties, bioactive potential and extraction processes of glycosaminoglycans: an overview

Investigation of sample handling steps for accurate heparan sulphate disaccharide analysis using HPLC–MS

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