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International Journal of Pharmaceutical Chemistry and Analysis


Stability Indicating RP-HPLC Method for In-Vitro Analysis of 5-Flurouracil


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Author Details: Md. Habban Akhter, Saima Amin, Showkat R Mir

Volume : 3

Issue : 1

Online ISSN : 2394-2797

Print ISSN : 2394-2789

Article First Page : 18

Article End Page : 24


Abstract

5-Flurouracil (FU), {5-fluoropyrimidines-2,4(1H,3H)-dione} is an anticancer drug that competitively inhibits thymidylate synthatase, an enzyme required for DNA synthesis that leads to programmed cell death of proliferating cells. It is used as topical cream for squamous cell carcinoma but a little information is available for its active drug targeting. Our attempt is to generate a validated stability indicating RP-HPLC method for the quantitation of 5-FU for active targeting to squamous cell carcinoma. In our study, RP-HPLC system used was (SHIMADZU (AT vp), Japan) and comprised of UV detector (SCL-10A vp), a pump system (LC-10 AT vp), and an injector. The separation was achieved on C18 LiChrospher®100 (250 x 4.6 mm, 5 μm) and elution was monitored gradiently with mobile phase consisting of HPLC water at a flow rate of 1ml/min. The injection volume was 20 µL previously filtered through 0.45μ membrane filter and elute was analyzed at the set wavelength of 269 nm. The proposed method was validated for various parameters such as specificity, linearity, precision, accuracy, and robustness. The calibration plot drawn was found to be linear over the concentration range of 1–10µg/mL and within the acceptable range (R2 = 0.999). The developed method was found to be simple, precise, and accurate and can be reproduced with LOD and LOQ of 0.007108 and 0.021539µg/mL respectively. The degradation study at variable temperatures such as 0oC; 8oC, 25oC, 37oC and pH 1.2, 3.2, 4.2, 6.8 and 7.4 after storage for 24 h suggested that 5-FU was stable at aforementioned temperatures and pH. It further signifies that the method is suitable for in-process quality control analysis of new composition of 5- FU due to good accuracy and high degree of precision.

Keywords:
5-Fluorouracil, Active targeting, Assay validation, RP-HPLC, Squamous cell carcinoma, Solubility