Determination of methotrexate and its metabolite 7-hydroxymethotrexate by direct injection of human plasma into a column-switching liquid chromatographic system using post-column photochemical reaction with fluorimetric detection

39Citations
Citations of this article
4Readers
Mendeley users who have this article in their library.
Get full text

Abstract

A simple, sensitive and fully automated column-switching system by direct injection of plasma samples for determination of methotrexate and its metabolite 7-hydroxymethotrexate was developed. The system utilized a C8 alkyl-diol silica precolumn coupled with a LiChrospher RP-18 analytical column, followed by a photoreactor and fluorimetric detection. The photo-oxidative irradiation was accomplished at UV 254 nm in the presence of 0.1% hydrogen peroxide in the eluent. Studies showed that the fluorimetric response was influenced by the reaction time, the degree of the reactor's transparency and the choice of the working wavelengths. By optimizing the content of acetonitrile in the eluent, methotrexate can be separated from 7-hydroxymethotrexate completely. The method validation revealed quantitative recoveries (≤94%) with coefficients of variation ≤4.4%. The limits of detection and quantitation for determination of methotrexate were 0.20 and 0.36 ng, respectively, corresponding to 2.0 and 3.6 ng/ml for an injection volume of 100 μl. It was possible to enhance the sensitivity further by injecting larger plasma volumes, up to 500 μl.

Cite

CITATION STYLE

APA

Yu, Z., Westerlund, D., & Boos, K. S. (1997). Determination of methotrexate and its metabolite 7-hydroxymethotrexate by direct injection of human plasma into a column-switching liquid chromatographic system using post-column photochemical reaction with fluorimetric detection. Journal of Chromatography B: Biomedical Applications, 689(2), 379–386. https://doi.org/10.1016/S0378-4347(96)00357-X

Register to see more suggestions

Mendeley helps you to discover research relevant for your work.

Already have an account?

Save time finding and organizing research with Mendeley

Sign up for free