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Separation of human fibrinopeptides by capillary zone electrophoresis.
Electrophoresis. 2005 Feb; 26(3):600-9.E

Abstract

We describe a method for the simultaneous determination of the five fibrinopeptide forms derived from the thrombin-promoted activation of human fibrinogen by capillary zone electrophoresis (CZE). The fibrinopeptide mixture was first desalted by a solid-phase extraction (SPE) step. The analysis was performed in reversed polarity in a highly cross-linked polyethylene glycol (PEG)-coated capillary with UV-light absorption detection at 200 nm. Several parameters including buffer concentration and pH, presence of an organic modifier, temperature, and applied voltage, have been tested. The best separations were obtained within 20 min, utilizing a 20 mM sodium phosphate buffer without organic modifier, in the narrow 6.1-6.2 pH range, at 25 degrees C, with an applied voltage of 20 kV. Quantitative analysis is made possible by the use of sheep fibrinopeptide A as an internal standard to correct for both extraction and injection errors.

Authors+Show Affiliations

Profumo A
S.C. Medicina Rigenerativa, Istituto Nazionale per la Ricerca sul Cancro, Genova, Italy. aldo.profumo@istge.it
Cardinali B
No affiliation info available
Cuniberti C
No affiliation info available
Rocco M
No affiliation info available

MeSH

AnimalsChromatography, High Pressure LiquidElectrophoresis, CapillaryFibrinopeptide AFibrinopeptide BHumansHydrogen-Ion ConcentrationReference StandardsReproducibility of ResultsSensitivity and SpecificitySheep

Pub Type(s)

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

Language

eng

PubMed ID

15690433

Citation

Profumo, Aldo, et al. "Separation of Human Fibrinopeptides By Capillary Zone Electrophoresis." Electrophoresis, vol. 26, no. 3, 2005, pp. 600-9.
Profumo A, Cardinali B, Cuniberti C, et al. Separation of human fibrinopeptides by capillary zone electrophoresis. Electrophoresis. 2005;26(3):600-9.
Profumo, A., Cardinali, B., Cuniberti, C., & Rocco, M. (2005). Separation of human fibrinopeptides by capillary zone electrophoresis. Electrophoresis, 26(3), 600-9.
Profumo A, et al. Separation of Human Fibrinopeptides By Capillary Zone Electrophoresis. Electrophoresis. 2005;26(3):600-9. PubMed PMID: 15690433.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Separation of human fibrinopeptides by capillary zone electrophoresis. AU - Profumo,Aldo, AU - Cardinali,Barbara, AU - Cuniberti,Carla, AU - Rocco,Mattia, PY - 2005/2/4/pubmed PY - 2005/6/10/medline PY - 2005/2/4/entrez SP - 600 EP - 9 JF - Electrophoresis JO - Electrophoresis VL - 26 IS - 3 N2 - We describe a method for the simultaneous determination of the five fibrinopeptide forms derived from the thrombin-promoted activation of human fibrinogen by capillary zone electrophoresis (CZE). The fibrinopeptide mixture was first desalted by a solid-phase extraction (SPE) step. The analysis was performed in reversed polarity in a highly cross-linked polyethylene glycol (PEG)-coated capillary with UV-light absorption detection at 200 nm. Several parameters including buffer concentration and pH, presence of an organic modifier, temperature, and applied voltage, have been tested. The best separations were obtained within 20 min, utilizing a 20 mM sodium phosphate buffer without organic modifier, in the narrow 6.1-6.2 pH range, at 25 degrees C, with an applied voltage of 20 kV. Quantitative analysis is made possible by the use of sheep fibrinopeptide A as an internal standard to correct for both extraction and injection errors. SN - 0173-0835 UR - https://www.unboundmedicine.com/medline/citation/15690433/Separation_of_human_fibrinopeptides_by_capillary_zone_electrophoresis_ DB - PRIME DP - Unbound Medicine ER -