Analytical method for keratan sulfates by high-performance liquid chromatography/turbo-ionspray tandem mass spectrometry

We established a highly sensitive LC/MS/MS method for the analysis of the d isaccharides produced from keratan sulfates (KS), It was revealed that the disaccharides produced by keratanase II enzymatic digestion of RS could be determined with high sensitivity by the negative-ion mode of multiple react ion monitoring. Furthermore,monosulfatedanddisulfateddisaccharides can be s eparated using a short column of Capcell Pak NH2 UG80 (35 mm x 2 mm i.d.). The complete analysis of one sample can be performed within 5 min. The assa y method was validated and showed satisfactory sensitivity, precision, and accuracy, which enabled quantitation at subpicomole levels. From the result s of analyses of KS obtained from cornea, nasal cartilage, and brain, it wa s found that the degree of sulfation at the C-6 position of the galactose r esidues differed among those samples in the following order: nasal cartilag e > cornea > brain. Our analytical method is very useful for the analyses o f KS in various biological materials and for comparison of the degree of su lfation of KS from various biological samples. (C) 2001 Academic Press.