Analytical Method for Keratan Sulfates by High-Performance Liquid Chromatography/Turbo-Ionspray Tandem Mass Spectrometry
Autor: | Toshio Imanari, Toshihiro Oguma, Toshihiko Toida, Hidenao Toyoda |
---|---|
Rok vydání: | 2001 |
Předmět: |
Quality Control
Chromatography Chemistry Selected reaction monitoring Biophysics Analytical chemistry Reproducibility of Results Cell Biology Disaccharides Tandem mass spectrometry Mass spectrometry Biochemistry High-performance liquid chromatography Biological materials Highly sensitive Cornea Sulfation Keratan Sulfate Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization Animals Cattle Proteoglycans Keratanase II Molecular Biology Chromatography High Pressure Liquid |
Zdroj: | Analytical Biochemistry. 290:68-73 |
ISSN: | 0003-2697 |
DOI: | 10.1006/abio.2000.4940 |
Popis: | We established a highly sensitive LC/MS/MS method for the analysis of the disaccharides produced from keratan sulfates (KS). It was revealed that the disaccharides produced by keratanase II enzymatic digestion of KS could be determined with high sensitivity by the negative-ion mode of multiple reaction monitoring. Furthermore, monosulfated and disulfated disaccharides can be separated using a short column of Capcell Pak NH 2 UG80 (35 mm × 2 mm i.d.). The complete analysis of one sample can be performed within 5 min. The assay method was validated and showed satisfactory sensitivity, precision, and accuracy, which enabled quantitation at subpicomole levels. From the results of analyses of KS obtained from cornea, nasal cartilage, and brain, it was found that the degree of sulfation at the C-6 position of the galactose residues differed among those samples in the following order: nasal cartilage > cornea > brain. Our analytical method is very useful for the analyses of KS in various biological materials and for comparison of the degree of sulfation of KS from various biological samples. |
Databáze: | OpenAIRE |
Externí odkaz: |
načítá se...