Tags

Type your tag names separated by a space and hit enter

A novel LIF-CE method for the separation of hyaluronan- and chondroitin sulfate-derived disaccharides: Application to structural and quantitative analyses of human plasma low- and high-charged chondroitin sulfate isomers.
Electrophoresis. 2007 Jul; 28(14):2439-47.E

Abstract

The report describes a rapid and simple CE method using LIF detection for the analysis of unsaturated disaccharides obtained from enzymatic depolymerization of plasma chondroitin sulfate (CS) isomers. The disaccharide reducing groups were labeled with 2-aminoacridone (AMAC). The fluorotagged products can be separated by reversed-polarity CE using a sodium acetate buffer, pH 3.8, in the presence of 0.05% methylcellulose. The choice of the appropriate electrophoretic conditions was performed after a deep analysis of the most important parameters affecting analyte separation. In particular, the effect of both run buffer concentration and pH on resolution, efficiency, migration times, and peak area was evaluated. The selected electrophoretic conditions allowed us to separate the CS isomers-derived Delta-disaccharides in less than 12 min, also resolving the nonsulfated disaccharides released from CS isomers from those released from hyaluronan (HA). Moreover, these conditions gave a good reproducibility of both the migration times (CV%, 0.25) and the peak areas (CV%, 1.4). Intra- and interassay CV were 5.37 and 7.23%, respectively, and analytical recovery was about 86%. The applicability of the above method to the quantitative and structural disaccharide analyses of plasma CS isomers was investigated. Data obtained from 44 healthy human subjects were compared with those obtained by a fluorophore-assisted carbohydrate electrophoresis (FACE) reference assay, by using the Passing and Bablok regression and Bland-Altman tests. The developed method could represent a good tool for an ultrasensitive analysis of CS isomers in biological samples from different sources, particularly when samples are available in very low amounts.

Authors+Show Affiliations

Zinellu A
Dipartimento di Scienze Biomediche, Cattedra di Biochimica Clinica, Università degli Studi di Sassari, Italia. angelozinellu@libero.it
Pisanu S
No affiliation info available
Zinellu E
No affiliation info available
Lepedda AJ
No affiliation info available
Cherchi GM
No affiliation info available
Sotgia S
No affiliation info available
Carru C
No affiliation info available
Deiana L
No affiliation info available
Formato M
No affiliation info available

MeSH

Chondroitin SulfatesDisaccharidesElectrophoresis, CapillaryHumansHyaluronic AcidIsomerism

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

17577197

Citation

Zinellu, Angelo, et al. "A Novel LIF-CE Method for the Separation of Hyaluronan- and Chondroitin Sulfate-derived Disaccharides: Application to Structural and Quantitative Analyses of Human Plasma Low- and High-charged Chondroitin Sulfate Isomers." Electrophoresis, vol. 28, no. 14, 2007, pp. 2439-47.
Zinellu A, Pisanu S, Zinellu E, et al. A novel LIF-CE method for the separation of hyaluronan- and chondroitin sulfate-derived disaccharides: Application to structural and quantitative analyses of human plasma low- and high-charged chondroitin sulfate isomers. Electrophoresis. 2007;28(14):2439-47.
Zinellu, A., Pisanu, S., Zinellu, E., Lepedda, A. J., Cherchi, G. M., Sotgia, S., Carru, C., Deiana, L., & Formato, M. (2007). A novel LIF-CE method for the separation of hyaluronan- and chondroitin sulfate-derived disaccharides: Application to structural and quantitative analyses of human plasma low- and high-charged chondroitin sulfate isomers. Electrophoresis, 28(14), 2439-47.
Zinellu A, et al. A Novel LIF-CE Method for the Separation of Hyaluronan- and Chondroitin Sulfate-derived Disaccharides: Application to Structural and Quantitative Analyses of Human Plasma Low- and High-charged Chondroitin Sulfate Isomers. Electrophoresis. 2007;28(14):2439-47. PubMed PMID: 17577197.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A novel LIF-CE method for the separation of hyaluronan- and chondroitin sulfate-derived disaccharides: Application to structural and quantitative analyses of human plasma low- and high-charged chondroitin sulfate isomers. AU - Zinellu,Angelo, AU - Pisanu,Salvatore, AU - Zinellu,Elisabetta, AU - Lepedda,Antonio J, AU - Cherchi,Gian Mario, AU - Sotgia,Salvatore, AU - Carru,Ciriaco, AU - Deiana,Luca, AU - Formato,Marilena, PY - 2007/6/20/pubmed PY - 2007/10/5/medline PY - 2007/6/20/entrez SP - 2439 EP - 47 JF - Electrophoresis JO - Electrophoresis VL - 28 IS - 14 N2 - The report describes a rapid and simple CE method using LIF detection for the analysis of unsaturated disaccharides obtained from enzymatic depolymerization of plasma chondroitin sulfate (CS) isomers. The disaccharide reducing groups were labeled with 2-aminoacridone (AMAC). The fluorotagged products can be separated by reversed-polarity CE using a sodium acetate buffer, pH 3.8, in the presence of 0.05% methylcellulose. The choice of the appropriate electrophoretic conditions was performed after a deep analysis of the most important parameters affecting analyte separation. In particular, the effect of both run buffer concentration and pH on resolution, efficiency, migration times, and peak area was evaluated. The selected electrophoretic conditions allowed us to separate the CS isomers-derived Delta-disaccharides in less than 12 min, also resolving the nonsulfated disaccharides released from CS isomers from those released from hyaluronan (HA). Moreover, these conditions gave a good reproducibility of both the migration times (CV%, 0.25) and the peak areas (CV%, 1.4). Intra- and interassay CV were 5.37 and 7.23%, respectively, and analytical recovery was about 86%. The applicability of the above method to the quantitative and structural disaccharide analyses of plasma CS isomers was investigated. Data obtained from 44 healthy human subjects were compared with those obtained by a fluorophore-assisted carbohydrate electrophoresis (FACE) reference assay, by using the Passing and Bablok regression and Bland-Altman tests. The developed method could represent a good tool for an ultrasensitive analysis of CS isomers in biological samples from different sources, particularly when samples are available in very low amounts. SN - 0173-0835 UR - https://www.unboundmedicine.com/medline/citation/17577197/A_novel_LIF_CE_method_for_the_separation_of_hyaluronan__and_chondroitin_sulfate_derived_disaccharides:_Application_to_structural_and_quantitative_analyses_of_human_plasma_low__and_high_charged_chondroitin_sulfate_isomers_ L2 - https://doi.org/10.1002/elps.200600668 DB - PRIME DP - Unbound Medicine ER -