Improved HPLC method for total plasma homocysteine detection and quantification.
Recent clinical research has pointed at hyperhomocysteinemia as an independent risk factor in a number of cardiovascular and neurological diseases. We have improved a chromatographic method of total plasma homocysteine measurements in order to obtain higher sensitivity, reliability and reproducibility. The method demonstrates excellent linearity (R=0.999), range (<2-100 microM), precision (instrumental RSD 0.06 and method RSD 1.17), accuracy (recovery of 99.92 and RSD 1.27), reproducibility, quantification limit and ruggedness (e.g. pH from 2.0 to 2.5). Because even a small increase in homocysteine level can be a significant risk factor of cardiovascular diseases, such a precise method is required. The constructed method allows the measurement of plasma pyridoxal phosphate, PLP, the co-enzyme form of vitamin B(6), on the same column and similar reagents. The developed method has been successfully applied to measure both total plasma and serum homocysteine in a group of acute stroke patients.
Sawuła
Wojciech
W
Department of Molecular and Cellular Biology, Intercollegiate Faculty of Biotechnology of University of Gdańsk and Medical University of Gdańsk, Poland.
Banecka-Majkutewicz
Zyta
Z
Kadziński
Leszek
L
Jakóbkiewicz-Banecka
Joanna
J
Wegrzyn
Grzegorz
G
Nyka
Walenty
W
Banecki
Bogdan
B
D001774:Blood Chemical Analysis / Q000379:methods*; D001777:Blood Coagulation; D002021:Buffers; D002624:Chemistry, Clinical / Q000379:methods; D002845:Chromatography / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005456:Fluorescent Dyes / Q000494:pharmacology; D006710:Homocysteine / Q000097:blood*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D015203:Reproducibility of Results; D025101:Vitamin B 6 / Q000737:chemistry; D014914:Whole Blood Coagulation Time
D002021:Buffers; D005456:Fluorescent Dyes; D006710:Homocysteine; D025101:Vitamin B 6