Simultaneous Method for Quantification of Genotoxic Impurity in the Gemcitabine Hydrochloride by RP-HPLC

: Objective: In the pharmaceutical analytical field, effort to analyzing the degradation or related and impurity substances from the complex matrix in the ultra-trace levels is a really a challengeable task. Impurity is any component of the new drug substance that is not the chemical entity defined as the new drug substance. Gemcitabine hydrochloride is a new anticancer bio molecule. According to Indian Pharmacopoeia there is two impurities present in this drug. The literature review exploded that, there is no specific impurity profiling based analytical method developed to estimate the Cytosine. Methods: RP HPLC is developed to quantify and separate Gemcitabine hydrochloride and Cytosine. The Hibar® C18 (250 x 4.6 mm i.d., 5μ), Column choices as stationary phase, tri ethyl ether and Acetonitrile in ratio of 95:5 with 4.35 (with Orthophosphoric acid) as mobile phase, the flow rate fixed as 1ml/ min. at 279 nm. Results: A retention time found to be 3.44min and 8.28min for gemcitabine and cytosine respectively. The developed method was validated as per ICH guidelines and the method was proved as selective, specific and linear. The assay and recovery studies were carried out the assay limit was 99.19 and 99.32 for bulk drug and formulations respectively. The Cytosine were present 0.7821 μg/ml and 0.6531 μg/ml in the bulk drug and formulation respectively. Conclusion: It can be concluded that the developed RP-HPLC method was found to be suitable for the appraisal of cytosine impurities in Gemcitabine HCl. The developed RP-HPLC is economic and suitable for the use.Key words: Gemcitabine HCl (GCH), Cytosine (CYS), RP-HPLC, Genotoxin, Impurity, Anticancer.