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Analytical method for keratan sulfates by high-performance liquid chromatography/turbo-ionspray tandem mass spectrometry.
Anal Biochem. 2001 Mar 01; 290(1):68-73.AB

Abstract

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 NH2 UG80 (35 mm x 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.

Authors+Show Affiliations

Faculty of Pharmaceutical Sciences, Chiba University, 1-33 Yayoi, Inage, Chiba, 263-8522, Japan.ogumab0g@daiichipharm.co.jpNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

11180938

Citation

Oguma, T, et al. "Analytical Method for Keratan Sulfates By High-performance Liquid Chromatography/turbo-ionspray Tandem Mass Spectrometry." Analytical Biochemistry, vol. 290, no. 1, 2001, pp. 68-73.
Oguma T, Toyoda H, Toida T, et al. Analytical method for keratan sulfates by high-performance liquid chromatography/turbo-ionspray tandem mass spectrometry. Anal Biochem. 2001;290(1):68-73.
Oguma, T., Toyoda, H., Toida, T., & Imanari, T. (2001). Analytical method for keratan sulfates by high-performance liquid chromatography/turbo-ionspray tandem mass spectrometry. Analytical Biochemistry, 290(1), 68-73.
Oguma T, et al. Analytical Method for Keratan Sulfates By High-performance Liquid Chromatography/turbo-ionspray Tandem Mass Spectrometry. Anal Biochem. 2001 Mar 1;290(1):68-73. PubMed PMID: 11180938.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Analytical method for keratan sulfates by high-performance liquid chromatography/turbo-ionspray tandem mass spectrometry. AU - Oguma,T, AU - Toyoda,H, AU - Toida,T, AU - Imanari,T, PY - 2001/2/22/pubmed PY - 2001/5/22/medline PY - 2001/2/22/entrez SP - 68 EP - 73 JF - Analytical biochemistry JO - Anal Biochem VL - 290 IS - 1 N2 - 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 NH2 UG80 (35 mm x 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. SN - 0003-2697 UR - https://www.unboundmedicine.com/medline/citation/11180938/Analytical_method_for_keratan_sulfates_by_high_performance_liquid_chromatography/turbo_ionspray_tandem_mass_spectrometry_ DB - PRIME DP - Unbound Medicine ER -