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Colorectal Cancer (CRC) is constantly increasing in incidence both worldwide and at the national level. Chemotherapeutic agents often prescribed in CRC are Capecitabine (CCB) and 5-Fluorouracil (FU). CCB is activated to FU in a three steps reaction giving 5'-deoxy-5-fluorocytidine (DFCR), followed by 5'-deoxy-5-fluorouridine (DFUR) to yield finally FU, the active form, which is later deactivated to 5,6-dihydro-5-fluorouracil (DHFU). Patients exhibited variable responses and adverse events in response to CCB therapy, despite being treated with the same dose. This could be explained by the presence of different possible enzyme SNPs that can occur along the CCB activation and deactivation pathways. This study aims at developing a new method of analysis of CCB and its metabolites using HPLC-UV, to determine the plasma concentrations of CCB and its metabolites DFCR, DFUR, FU, DHFU and 5-Chlorouracil (CLU; the internal standard), followed by a correlation study with the toxicities occurring during therapy, to become a predictive method for toxicity, away from the exhausting genotyping process. A new superior analytical method is presented using computer-assisted method development, which achieved full separation of the six analytes during the least possible gradient time, eluting the compounds at 2.8, 3.2, 4.4, 5.2, 5.8 and 9.9 minutes for DHFU, FU, CLU, DFCR, DFUR and CCB, respectively. The method showed accuracy, precision and robustness upon validation. Clinical results showed a positive correlation between the DFCR concentration and mucositis, as well as, between the DFUR concentration and Hand-Foot Syndrome, confirming that this technique could be used for predicting such toxicities in CRC patients.

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