A validated stability-indicating HPTLC method for the estimation of gemcitabine HCl in its dosage form

Abstract

Gemcitabine belongs to a class of drugs known as antimetabolites (pyrimidine analog) used in various carcinomas, such as pancreatic cancer, bladder cancer, and breast cancer. The present study deals with the development and validation of a stability-indicating highperformance thin-layer chromatography (HPTLC) method for the estimation of gemcitabine using TLC plates precoated with silica gel 60 F254 as stationary phase and toluene-methanol-chloroform in the ratio of 3.6:3.6:3 (v/v/v) as the mobile phase. Gemcitabine (RF 0.48 ± 0.03) and its degradation products were well resolved. The wavelength selected for quantitation was 268 nm. The method was linear in the concentration range 500–3000 ng/band with a correlation coefficient of 0.997. The repeatability for six samples was 1.88%RSD. The intraday and interday precisions were 1.35–1.60%RSD and 1.57–1.81%RSD, respectively. The accuracy (recovery) was found to be in the range 98.30–101.75%. The drug was subjected to stress conditions such as hydrolysis, oxidation, photolysis, and heat. Degradation products produced as a result of the stress conditions did not interfere with the detection of gemcitabine; therefore the proposed method can be considered stability-indicating. Gemcitabine did not degrade under thermal and photolytic conditions but showed degradation under hydrolytic and oxidative conditions. The developed method was found to be simple, specific, precise, and stability-indicating.