RP-HPLC Method for the Quantification of Impurities in Etelcalcetide Injection and a Comparative Characterization Study of In-House Samples of Etelcalcetide Injection with Reference-Listed Drug Samples

In present study, an accurate and simple reversed phase-high performance liquid chromatography (RP-HPLC) method for the determination of potential impurities resulting from etelcalcetide injection was validated. An Ace Excel 3 C18 amide column with 100% 0.1 M sodium perchlorate (pH 2.0) buffer (100%) was used as mobile phase A and a 60:10 (%v/v) ratio of acetonitrile and 0.1 M sodium perchlorate (pH 2.0) was used as mobile phase B. The wavelength detection performed at 210 nm. Etelcalcetide injection was subjected to thermal, photolytic, acid, base, and peroxide degradation conditions, which were studied using the current methodology. The method was validated as per the International Council for Harmonization guideline (ICH). Consistent recoveries were obtained for all the impurities of etelcalcetide between 85% and 105%. Furthermore, comparative characterization studies and impurity profiling were successfully performed for in-house samples of etelcalcetide with reference-listed drug (RLD) samples through circular dichroism (CD), Fourier transform infrared (FTIR), size exclusion chromatography-high performance liquid chromatography (SEC-HPLC) and liquid chromatography-high resolution mass spectrometry (LC-HRMS), results indicates that found similar in both in-house and RLD samples.