A NEW HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY METHOD FOR THE SEPARATION AND SIMULTANEOUS QUANTIFICATION OF EPTIFIBATIDE AND ITS IMPURITIES IN PHARMACEUTICAL INJECTION FORMULATION
Objective: The objective of the present study is to develop a stability indicating RP-HPLC method for qualitative and quantitative determination of Eptifibatide in bulk and dosage form and in presence of its impurities 1 and 2.
Method: The chromatographic separation was carried out on Phenomenex Luna C18 column (250mm×4.6mm; 5µ id) as stationary phase, methanol and phosphate buffer at pH 6.4 in the ratio of 65:45 (v/v) as mobile phase at a flow rate of 1.0 mL/min, UV detection was carried at wavelength of 236 nm and the analysis was completed with a run time of 15 min.
Result: In the developed conditions the retention time of Eptifibatide and its impurities 1 and 2 was found to be 3.35, 4.93 and 8.18 min respectively. The method was validated for system suitability, range of analysis, precision, specificity, stability and robustness. Spiked recovery at 50 %, 100 % and 150 % was carried for both standard and impurities and the acceptable % recovery of 98-102 was observed for Eptifibatide and both impurities studied and the % RSD in each spiked level was found to be less than 2. Stability tests were done through exposure of the analyte solution to five different stress conditions i.e expose to 1N HCl, 1 N NaOH, 3 % H2O2, 800C temperature to UV radiation. In all the degradation condition, standard drug Eptifibatide was detected along with both the impurities studied and the degradation products were successfully separated. In the formulation analysis there is no other chromatographic detection of other impurities and formulation excipients.
Conclusion: The developed method was found to be suitable for the quantification of Eptifibatide and can separate and analyse impurities 1 and 2.
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