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Revista de Chimie (Rev. Chim.), Year 2015, Volume 66, Issue 5,





Carolina Negrei, Constantin CAruntu, Octav Ginghina, George Traian Alexandru Burcea Dragomiroiu, Corina Dalia Toderescu, Daniel Boda
Qualitative and Quantitative Determination of Methotrexate Polyglutamates in Erythrocytes By High Performance Liquid Chromatography

Abstract:

Methotrexate (MTX) is a structural folic acid analogue. It is one of the most easily tolerated antipsoriatic drugs with disease-modifying capacities and a prodrug which is activated to yield methotrexate polyglutamates (MTX-PG) by means of folylpolyglutamate synthase (FP-GS). MTX-PGs concentration in the tissues is low and the quantification of individual MTX-PG species is difficult. A HPLC method for the quantification of these metabolites was developed. The MTXPGs were extracted from the biological matrix using 100 µl red blood cell hemolysate, by perchloric acid deproteinization step with direct injection of the extract into the HPLC. We used a C18 reversed-phase column, an acetonitrile buffer, and postcolumn photo-oxidation of MTXPGs to fluorescent analytes. The complete conversion to MTX of the MTX-PG2-7 pool, the incubation was performed in the dark, at 37 °C. The correlation coefficients was ]0.995 for MTX-PG1 as well as for samples of MTX-PG1 obtained following enzymatic conversion of MTX-PG 2-7. Intra- and interday imprecision were [10%. The advantage of this method lies in its simplicity and low volume of RBC required. Keywords: HPLC, methotrexate, methotrexate polyglutamates


Issue: 2015, Volume 66, Issue 5
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