Abstract :
Aim: The aim of this study is to develop a new stability indicating reversed-phase high-performance liquid
chromatography (RP-HPLC) method for the simultaneous estimation of Tegafur (TEG) and Uracil (URA) in capsule
dosage form. Materials and Methods: Chromatography was carried out on a C18 Phenomenex column (250 mm ×
4.6 mm i.d., 5 μm particle size) with an isocratic mobile phase composed of acetate buffer (pH 4) and methanol in
the proportion of 70:30 (v/v). The compounds were eluted at a steady flow rate of 1.0 mL/min with a total run time
of 10 min. The detection was carried out by ultraviolet-detector at 280 nm. The developed method was validated
as per the ICH guidelines, and stability studies were performed. Results and Discussion: TEG retention time was
about 5.2 min, and URA was 2.9 min. A calibration curve was obtained in the range of 25–125 μg/mL for TEG and
20–100 μg/mL for URA. Specificity experiments revealed the absence of interference from excipients. The mean
assay for TEG was found to be 101.4% (w/w) and for URA was 102.8% (w/w). The method was also evaluated for
robustness and ruggedness, and the results obtained were satisfactory. Forced degradation studies were performed
for TEG and URA in acidic and alkaline, thermal, and oxidative conditions, and no substantial degradation was
observed in all the conditions. Conclusion: It is concluded that the developed stability indicating RP-HPLC method
was specific, precise, accurate, sensitive, and robust for the simultaneous estimation of TEG and URA in capsules.
Keyword :
Reversed-phase high-performance liquid chromatography, stability indicating assay, Tegafur, Uracil, validation