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Analytical strategy for the assessment of the protein glycation status in uremic patients by high-performance liquid chromatography.

Abstract

We propose a newly integrated procedure for the analysis of furosine (early glycation product) and pentosidine (glycoxidation end-product) in plasma proteins and the simultaneous assessment of advanced glycation end-product (AGE) peptides and free pentosidine in plasma. In order to determine furosine and protein-linked pentosidine, plasma proteins were hydrolyzed in 8 M HCl and each analyte was purified by solid-phase extraction. Furosine was determined by ion-pair RP-HPLC methodology with isocratic elution and spectrophotometric detection at 280 nm and pentosidine by ion-pair RP-HPLC by using gradient elution and fluorimetric detection at 335/385 nm. To assess free pentosidine concentration and simultaneously evaluate the AGE peptides, an aliquot of plasma sample was diluted and ultrafiltered by using Centricon 10 M(r) < or = 10,000) ultrafiltration membranes. Free pentosidine and AGE peptides were analysed by ion-pair RP-HPLC, by using gradient elution and fluorimetric detection at 385 nm upon excitation at 335 nm. The HPLC methodology has been successfully used for the determination of glycation and glycoxidation protein status in uremic patients.

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  • Authors+Show Affiliations

    ,

    Dipartimento di Biologia Cellulare e Molecolare, Università di Perugia, Italy. floridi@unipg.it

    , ,

    Source

    Journal of chromatography. A 846:1-2 1999 Jun 18 pg 65-71

    MeSH

    Arginine
    Blood Proteins
    Chromatography, High Pressure Liquid
    Glucose
    Glycation End Products, Advanced
    Humans
    Lysine
    Peptides
    Reference Standards
    Spectrometry, Fluorescence
    Spectrophotometry, Ultraviolet
    Uremia

    Pub Type(s)

    Journal Article

    Language

    eng

    PubMed ID

    10420599

    Citation

    Floridi, A, et al. "Analytical Strategy for the Assessment of the Protein Glycation Status in Uremic Patients By High-performance Liquid Chromatography." Journal of Chromatography. A, vol. 846, no. 1-2, 1999, pp. 65-71.
    Floridi A, Trizza V, Paolotti P, et al. Analytical strategy for the assessment of the protein glycation status in uremic patients by high-performance liquid chromatography. J Chromatogr A. 1999;846(1-2):65-71.
    Floridi, A., Trizza, V., Paolotti, P., & Lucarelli, C. (1999). Analytical strategy for the assessment of the protein glycation status in uremic patients by high-performance liquid chromatography. Journal of Chromatography. A, 846(1-2), pp. 65-71.
    Floridi A, et al. Analytical Strategy for the Assessment of the Protein Glycation Status in Uremic Patients By High-performance Liquid Chromatography. J Chromatogr A. 1999 Jun 18;846(1-2):65-71. PubMed PMID: 10420599.
    * Article titles in AMA citation format should be in sentence-case
    TY - JOUR T1 - Analytical strategy for the assessment of the protein glycation status in uremic patients by high-performance liquid chromatography. AU - Floridi,A, AU - Trizza,V, AU - Paolotti,P, AU - Lucarelli,C, PY - 1999/7/27/pubmed PY - 1999/7/27/medline PY - 1999/7/27/entrez SP - 65 EP - 71 JF - Journal of chromatography. A JO - J Chromatogr A VL - 846 IS - 1-2 N2 - We propose a newly integrated procedure for the analysis of furosine (early glycation product) and pentosidine (glycoxidation end-product) in plasma proteins and the simultaneous assessment of advanced glycation end-product (AGE) peptides and free pentosidine in plasma. In order to determine furosine and protein-linked pentosidine, plasma proteins were hydrolyzed in 8 M HCl and each analyte was purified by solid-phase extraction. Furosine was determined by ion-pair RP-HPLC methodology with isocratic elution and spectrophotometric detection at 280 nm and pentosidine by ion-pair RP-HPLC by using gradient elution and fluorimetric detection at 335/385 nm. To assess free pentosidine concentration and simultaneously evaluate the AGE peptides, an aliquot of plasma sample was diluted and ultrafiltered by using Centricon 10 M(r) < or = 10,000) ultrafiltration membranes. Free pentosidine and AGE peptides were analysed by ion-pair RP-HPLC, by using gradient elution and fluorimetric detection at 385 nm upon excitation at 335 nm. The HPLC methodology has been successfully used for the determination of glycation and glycoxidation protein status in uremic patients. SN - 0021-9673 UR - https://www.unboundmedicine.com/medline/citation/10420599/Analytical_strategy_for_the_assessment_of_the_protein_glycation_status_in_uremic_patients_by_high_performance_liquid_chromatography_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0021-9673(99)00442-2 DB - PRIME DP - Unbound Medicine ER -