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Determination of homocysteine in human plasma by micellar electrokinetic chromatography and in-capillary detection reaction with 2,2'-dipyridyl disulfide.
J Chromatogr A. 2003 Mar 21; 990(1-2):197-204.JC

Abstract

We present a new method for homocysteine quantitation in human plasma based on in-capillary reaction of homocysteine with 2,2'-dipyridyl disulfide. Homocysteine is in this so-called thiol-exchange reaction quantitatively transformed in mixed disulfide concomitantly with formation of an equimolar amount of 2-thiopyridone that is further separated by micellar electrokinetic chromatography and determined specifically at 343 nm. The concentration of homocysteine is thus estimated indirectly from the result of 2-thiopyridone determination. The linear detection range for concentration versus peak area for the assay was from 0.03-3 mM (correlation coefficient 0.994) with a detection limit of 6 microM and a limit of quantitation 20 microM. The inter-day reproducibility of the peak area and the migration time were 1.37% and 0.05%, respectively. The method is simple, relatively rapid and can be easily automated. Moreover the common capillary electrophoresis apparatus with a UV detector can be used to distinguish between normal and pathological hyperhomocysteinemia plasma samples.

Authors+Show Affiliations

Department of Biochemistry, Faculty of Science, Masaryk University, Kotlárská 2, 611 37 Brno, Czech Republic.No affiliation info availableNo affiliation info available

Pub Type(s)

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

Language

eng

PubMed ID

12685598

Citation

Sevcíková, Petra, et al. "Determination of Homocysteine in Human Plasma By Micellar Electrokinetic Chromatography and In-capillary Detection Reaction With 2,2'-dipyridyl Disulfide." Journal of Chromatography. A, vol. 990, no. 1-2, 2003, pp. 197-204.
Sevcíková P, Glatz Z, Tomandl J. Determination of homocysteine in human plasma by micellar electrokinetic chromatography and in-capillary detection reaction with 2,2'-dipyridyl disulfide. J Chromatogr A. 2003;990(1-2):197-204.
Sevcíková, P., Glatz, Z., & Tomandl, J. (2003). Determination of homocysteine in human plasma by micellar electrokinetic chromatography and in-capillary detection reaction with 2,2'-dipyridyl disulfide. Journal of Chromatography. A, 990(1-2), 197-204.
Sevcíková P, Glatz Z, Tomandl J. Determination of Homocysteine in Human Plasma By Micellar Electrokinetic Chromatography and In-capillary Detection Reaction With 2,2'-dipyridyl Disulfide. J Chromatogr A. 2003 Mar 21;990(1-2):197-204. PubMed PMID: 12685598.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Determination of homocysteine in human plasma by micellar electrokinetic chromatography and in-capillary detection reaction with 2,2'-dipyridyl disulfide. AU - Sevcíková,Petra, AU - Glatz,Zdenek, AU - Tomandl,Josef, PY - 2003/4/11/pubmed PY - 2003/12/3/medline PY - 2003/4/11/entrez SP - 197 EP - 204 JF - Journal of chromatography. A JO - J Chromatogr A VL - 990 IS - 1-2 N2 - We present a new method for homocysteine quantitation in human plasma based on in-capillary reaction of homocysteine with 2,2'-dipyridyl disulfide. Homocysteine is in this so-called thiol-exchange reaction quantitatively transformed in mixed disulfide concomitantly with formation of an equimolar amount of 2-thiopyridone that is further separated by micellar electrokinetic chromatography and determined specifically at 343 nm. The concentration of homocysteine is thus estimated indirectly from the result of 2-thiopyridone determination. The linear detection range for concentration versus peak area for the assay was from 0.03-3 mM (correlation coefficient 0.994) with a detection limit of 6 microM and a limit of quantitation 20 microM. The inter-day reproducibility of the peak area and the migration time were 1.37% and 0.05%, respectively. The method is simple, relatively rapid and can be easily automated. Moreover the common capillary electrophoresis apparatus with a UV detector can be used to distinguish between normal and pathological hyperhomocysteinemia plasma samples. SN - 0021-9673 UR - https://www.unboundmedicine.com/medline/citation/12685598/Determination_of_homocysteine_in_human_plasma_by_micellar_electrokinetic_chromatography_and_in_capillary_detection_reaction_with_22'_dipyridyl_disulfide_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0021-9673(03)00048-7 DB - PRIME DP - Unbound Medicine ER -