An HPLC method for the measurement of 5-fluorouracil in human plasma with a low detection limit and a high extraction yield

  • Authors:
    • Mark Adel Nassim
    • Farshad H. Shirazi
    • Christine M. Cripps
    • Shereeni Veerasinghan
    • Matshela J. Molepo
    • Mihail Obrocea
    • Diedre Redmond
    • Susan Bates
    • Diane Fry
    • David J. Stewart
    • Rakesh Goel
  • View Affiliations

  • Published online on: October 1, 2002     https://doi.org/10.3892/ijmm.10.4.513
  • Pages: 513-516
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Abstract

High performance liquid chromatographic (HPLC) techniques for the quantification of 5-fluorouracil (5-FU) in human plasma have been reported in the literature, however, a low limit of detection was generally found to result in a comparatively low extraction yield. We have developed a simple, rapid and sensitive HPLC method for the measurement of 5-FU in plasma which provides both a low limit of quantification and a high extraction yield. This method involves the solid phase extraction of 5-FU from a 500 μl plasma sample. The extract is then injected into an HPLC system equipped with a C18 μBondapak column, and a UV detector set at 260 nm. Ethyl acetate and potassium dihydrogen phosphate are used for the solid phase extraction and the HPLC mobile phase, respectively. This method provides in a good baseline, a sharp and symmetrical peak for 5-FU, and a high resolution between 5-FU and the internal standard. The retention time of 5-FU using this method is 4.7 min with a limit of detection of 5 ng/ml, and an extraction yield of 96.2±0.5% (SE). The next injection is possible in 11 min, and the coefficients of variation are 4.2-8.9% for interday precision, and 5.2-10.6% for day-to-day reproducibility. An HPLC method has been developed that has a low limit of detection and a high extraction yield. This technique was successfully applied in a clinical pharmacokinetic study of 5-FU.

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October 2002
Volume 10 Issue 4

Print ISSN: 1107-3756
Online ISSN:1791-244X

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Spandidos Publications style
Nassim MA, Shirazi FH, Cripps CM, Veerasinghan S, Molepo MJ, Obrocea M, Redmond D, Bates S, Fry D, Stewart DJ, Stewart DJ, et al: An HPLC method for the measurement of 5-fluorouracil in human plasma with a low detection limit and a high extraction yield. Int J Mol Med 10: 513-516, 2002
APA
Nassim, M.A., Shirazi, F.H., Cripps, C.M., Veerasinghan, S., Molepo, M.J., Obrocea, M. ... Goel, R. (2002). An HPLC method for the measurement of 5-fluorouracil in human plasma with a low detection limit and a high extraction yield. International Journal of Molecular Medicine, 10, 513-516. https://doi.org/10.3892/ijmm.10.4.513
MLA
Nassim, M. A., Shirazi, F. H., Cripps, C. M., Veerasinghan, S., Molepo, M. J., Obrocea, M., Redmond, D., Bates, S., Fry, D., Stewart, D. J., Goel, R."An HPLC method for the measurement of 5-fluorouracil in human plasma with a low detection limit and a high extraction yield". International Journal of Molecular Medicine 10.4 (2002): 513-516.
Chicago
Nassim, M. A., Shirazi, F. H., Cripps, C. M., Veerasinghan, S., Molepo, M. J., Obrocea, M., Redmond, D., Bates, S., Fry, D., Stewart, D. J., Goel, R."An HPLC method for the measurement of 5-fluorouracil in human plasma with a low detection limit and a high extraction yield". International Journal of Molecular Medicine 10, no. 4 (2002): 513-516. https://doi.org/10.3892/ijmm.10.4.513