Development and Validation of an HPLC-DAD Method for the Determination of 5-Fluorouracil in Plasma
Main Article Content
Abstract
Cancer remains one of the most significant health challenges worldwide, characterized by uncontrolled cell growth. However, it can be cured by using anticancer drugs. 5-fluorouracil is a drug used to treat various types of cancer with the inhibitory mechanism of the enzyme thymidylate synthase, which is crucial for DNA replication in cancer cells, leading to errors in their growth. Although effective, the short half-life of 5-FU and individual metabolic differences among patients limit its therapeutic potential. Therefore, it is necessary to monitor the 5-FU concentration during treatment to ensure the treatment efficiency. However, monitoring this compound in plasma is challenging due to the coexistence of complicated matrices. The study optimized and validated a sensitive method to quantify 5-fluorouracil in mice plasma using liquid-liquid extraction combined with a high-performance liquid chromatography-photodiode array detector (HPLC-PAD). The technique achieved excellent linearity, precision, accuracy, detection limits (LOD = 1.12 μg/L), and quantification (LOQ = 3.74μg/L). The optimal extraction solvent was ethyl acetate in two replicates (2x3ml), PSA/C18 was the most effective cleaning agent with 97.9 ± 0.2% recovery and the mass of PSA/C18 did not affect the extraction recovery. However, high plasma volume reduced the recovery of 5-FU in plasma. Finally, in vivo studies in mice were used to confirm this method. The findings of this research underscored the importance of personalized dosing strategies based on 5-FU concentration in blood.
Keywords
5-Fluorouracil (5-FU), HPLC-DAD, plasma, PSA, C18, pharmacokinetics
Article Details
References
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