Journal
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
Volume 413, Issue 7, Pages 1779-1785Publisher
SPRINGER HEIDELBERG
DOI: 10.1007/s00216-021-03152-7
Keywords
Chondroitin-sulfate; Glycosaminoglycan; High-performance liquid chromatography; Mass spectrometry; Stability; Recovery
Funding
- ELKH Research Centre for Natural Sciences
- National Research, Development and Innovation Office [OTKA PD 121187, OTKA FK 131603]
- Janos Bolyai Research Scholarship of the Hungarian Academy of Sciences
- National Research, Development and Innovation Fund of Hungary [2018-1.2.1-NKP-2018-00005, 2018-1.2.1-NKP]
- Hungarian Ministry of Human Resources [NTP-NFTO-20-B-0203]
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Chondroitin sulfate (CS) is a widely studied class of glycosaminoglycans, commonly analyzed based on disaccharide analysis; vacuum evaporation is preferred over lyophilization for solvent exchange, with higher recovery in aqueous solutions; proper storage and handling conditions are essential to prevent decomposition of the resulting disaccharide mixture.
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 degrees 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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