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The Development of a Rapid, Simple and Sensitive LC-MS/MS Method, to Guide Clinical Dosing, for the Analysis of 5-Fluorouracil in Human Plasma

[ Vol. 13 , Issue. 4 ]


Wenxuan He*, Jennifer H Martin, P Nicholas Shaw, Euan T Walpole and Goce Dimeski   Pages 378 - 383 ( 6 )


Introduction: 5-Fluorouracil is a commonly prescribed antineoplastic drug. Historically, methods to determine this compound in biological fluids have been time consuming or not sensitive enough.

Materials and Methods: In this report, a method using simple and rapid sample preparation with good sensitivity, precision and accuracy for the quantitative determination of 5-fluorouracil in human plasma by LC-MS/MS is established and validated. By the judicious combination of a small amount of injected sample, low flow rate, careful control of gradient elution profile, use of a HILIC pre-column to further remove residual phospholipids and an isotopically labelled internal standard (ILIS), the method has successfully overcome matrix effect with protein precipitation sample pre-treatment without evaporating to dryness and reconstituting process.

Conclusion: This newly developed method can be used to measure patient plasma concentrations of 5- fluorouracil, as low as 0.8 ng/mL and as high as 100 µ g/mL, with less than 15% both intra- and interday accuracy and precision. Such a methodology will be very convenient of being used in routine therapeutic drug monitoring.


5-Fluorouracil, LC-MS/MS, human plasma, protein precipitation, antineoplastic drug, sample preparation technique.


Department of Chemistry and Chemical Engineering, The University of Minjiang, Fuzhou, P.R., School of Pharmacy, The University of Queensland, Brisbane, School of Pharmacy, The University of Queensland, Brisbane, Princess Alexandra Hospital, Brisbane, Princess Alexandra Hospital, Brisbane

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