Skip to main content
Journal cover image

Expedient methodology for total methotrexate polyglutamation pool determination in human erythrocytes

Publication ,  Journal Article
Haandel, LV; Heemskerk, AAM; Becker, ML; Leeder, JS; Stobaugh, JF
Published in: Analytical Methods
July 1, 2010

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%. © 2010 The Royal Society of Chemistry.

Duke Scholars

Published In

Analytical Methods

DOI

EISSN

1759-9679

ISSN

1759-9660

Publication Date

July 1, 2010

Volume

2

Issue

7

Start / End Page

831 / 836

Related Subject Headings

  • 4004 Chemical engineering
  • 3401 Analytical chemistry
  • 0399 Other Chemical Sciences
  • 0301 Analytical Chemistry
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Haandel, L. V., Heemskerk, A. A. M., Becker, M. L., Leeder, J. S., & Stobaugh, J. F. (2010). Expedient methodology for total methotrexate polyglutamation pool determination in human erythrocytes. Analytical Methods, 2(7), 831–836. https://doi.org/10.1039/c0ay00164c
Haandel, L. V., A. A. M. Heemskerk, M. L. Becker, J. S. Leeder, and J. F. Stobaugh. “Expedient methodology for total methotrexate polyglutamation pool determination in human erythrocytes.” Analytical Methods 2, no. 7 (July 1, 2010): 831–36. https://doi.org/10.1039/c0ay00164c.
Haandel LV, Heemskerk AAM, Becker ML, Leeder JS, Stobaugh JF. Expedient methodology for total methotrexate polyglutamation pool determination in human erythrocytes. Analytical Methods. 2010 Jul 1;2(7):831–6.
Haandel, L. V., et al. “Expedient methodology for total methotrexate polyglutamation pool determination in human erythrocytes.” Analytical Methods, vol. 2, no. 7, July 2010, pp. 831–36. Scopus, doi:10.1039/c0ay00164c.
Haandel LV, Heemskerk AAM, Becker ML, Leeder JS, Stobaugh JF. Expedient methodology for total methotrexate polyglutamation pool determination in human erythrocytes. Analytical Methods. 2010 Jul 1;2(7):831–836.
Journal cover image

Published In

Analytical Methods

DOI

EISSN

1759-9679

ISSN

1759-9660

Publication Date

July 1, 2010

Volume

2

Issue

7

Start / End Page

831 / 836

Related Subject Headings

  • 4004 Chemical engineering
  • 3401 Analytical chemistry
  • 0399 Other Chemical Sciences
  • 0301 Analytical Chemistry