NDLI: Expedient methodology for total methotrexate polyglutamation pool determination in human erythrocytes

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Analytical Methods Vol - 2 Issue - 6

Analytical Methods Vol - 2 Issue - 5

Analytical Methods Vol - 2 Issue - 4

Analytical Methods Vol - 2 Issue - 3

Analytical Methods Vol - 2 Issue - 2

Analytical Methods Vol -1

Expedient methodology for total methotrexate polyglutamation pool determination in human erythrocytes

Content Provider	Royal Society of Chemistry (RSC)
Author	Stobaugh, John F. Heemskerk, Anthonius A. M. Haandel, Leon van Leeder, J. Steven Becker, Mara L.
Copyright Year	2010
Abstract	The measurement of methotrexate polyglutamate metabolites in red blood cells has potential to aid in individualization of methotrexate therapy in rheumatoid arthritis and juvenile idiopathic arthritis. In this report a method is presented for rapid analysis of these metabolites in human red blood cells. The analytical procedure is a simple “one pot” pre-column reaction involving the addition of sodium dithionite as a reducing agent followed by 15 minutes of boiling. After centrifugation the supernatant is introduced into a conventional HPLC system equipped with a fluorescence detector, without the need for further workup. By performing the derivatization reaction pre-column, a time consuming (6–14 h) commonly used deglutamation procedure utilizing blank human plasma, becomes obsolete. Using the described procedure the total sample preparation time for a 50 sample run should not exceed 1–1.5 hours. The chromatographic run time per sample is 7 minutes using isocratic elution conditions. The method was found to be linear over the clinical relevant concentration range of 10–500 nM of intra-cellular methotrexate polyglutamates. The intra-run mean accuracy of the target value was between 98.1% and 106.0%. The intra-run precision was between 1.2% and 8.8%.
Starting Page	831
Ending Page	836
Page Count	6
File Format	HTM / HTML PDF
ISSN	17599660
Volume Number	2
Issue Number	7
Journal	Analytical Methods
DOI	10.1039/c0ay00164c
Language	English
Publisher	Royal Society of Chemistry
Access Restriction	Open
Subject Keyword	Rheumatoid arthritis Centrifugation Fluorescence Sodium dithionite Methotrexate Reducing agent High-performance liquid chromatography Chromatography Juvenile idiopathic arthritis Blood plasma
Content Type	Text
Resource Type	Article
Subject	Analytical Chemistry Engineering Chemical Engineering

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