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Application of HPLC to study the kinetics of a branched bi-enzyme system consisting of hypoxanthine-guanine phosphoribosyltransferase and xanthine oxidase—an important biochemical system to evaluate the efficiency of the anticancer drug 6-mercaptopurine in ALL cell line by Sukirti Kalra; Manash K. Paul; Hemalatha Balaram; Anup Kumar Mukhopadhyay is a Chemistry article available to read on EtoBox.

What is Application of HPLC to study the kinetics of a branched bi-enzyme system consisting of hypoxanthine-guanine phosphoribosyltransferase and xanthine oxidase—an important biochemical system to evaluate the efficiency of the anticancer drug 6-mercaptopurine in ALL cell line about?

The thiopurine antimetabolite 6-mercaptopurine (6MP) is an important chemotherapeutic drug in the conventional treatment of childhood acute lymphoblastic leukemia (ALL). 6MP is mainly catabolized by both hypoxanthine-guanine phosphoribosyltransferase (HGPRT) and xanthine oxidase (XOD) to form thioinosinic monophosphate (TIMP) (therapeutically active metabolite) and 6-thiouric acid (6TUA) (inactive metabolite), respectively. The activity of both the enzymes varies among ALL patients governing the active and the inactive metabolite profile within the immature lymphocytes. Therefore, an attempt was made to study the kinetic nature of the branched bi-enzyme system acting on 6MP and to quantitate TIMP and 6TUA formed when the two enzymes are present in equal and variable ratios. The quantification of the branched kinetics using spectrophotometric method presents problem due to the closely apposed lambda(max) of the substrates and products. Hence, employing an HPLC method, the quantification of the products was done with the progress of time. The limit of quantification (LOQ) of substrate was found to be 10nM and for products as 50 nM. The limit of detection (LOD) was found to be 1 nM fo

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Author

Sukirti Kalra; Manash K. Paul; Hemalatha Balaram; Anup Kumar Mukhopadhyay

Publisher

Elsevier Science; Elsevier ; Elsevier BV (ISSN 1873-376X)

Published

2007

Language

EN

Field

Chemistry (Physical Sciences)