High‐performance liquid chromatography analysis of chondroitin sulphate isomers in human whole saliva in a variety of clinical conditions

OBJECTIVES: Tests have been carried out to assess the level of unsaturated disaccharide isomers obtained from chondroitin sulphate in whole saliva, which contains chondroitin sulphate derived from gingival crevicular fluid (GCF). MATERIALS AND METHODS: Whole saliva was collected from periodontally diseased subjects (PDS), clinically healthy subjects (CHS) and edentulous subjects (ES). Glycosaminoglycans (GAG) were liberated by digestion with Pronase E, and precipitated with cetylpyridinium chloride and ethanol. The unsaturated disaccharides obtained by chondroitinase ACII digestion of the liberated GAG were analysed by high-performance liquid chromatography. The unsaturated disaccharides included ΔDi-0S, ΔDi-6S and ΔDi-4S. RESULTS AND CONCLUSIONS: Analysis of data indicated that ΔDi-0S, ΔDi-6S and ΔDi-4S were found in all PDS samples. The amount (ng ml-1 collected whole saliva) of ΔDi-0S, ΔDi-6S and ΔDi-4S (P < 0.01) indicated significant differences between CHS and PDS whole saliva samples. The quantities of ΔDi-0S and ΔDi-4S (P < 0.01) indicated significant differences between PDS and ES whole saliva. The amount of ΔDi-0S (P < 0.05) and ΔDi-6S (P<0.01) also indicated significant differences between CHS and ES whole saliva. These results indicate that chondroitin sulphate in PDS and CHS whole saliva is representative of that previously reported in gingival crevicular fluid and so provides a useful and alternative means of assessing the role of GAG as indicators of periodontal disease.