Zaporožskij Medicinskij Žurnal (Oct 2018)
Validation of the method for determination of related impurities in the active antiviral ingredient of enisamium iodide
Abstract
The aim of the work. To validate a method for the related impurities determination in the enisamium iodide active pharmaceutical ingredient (API) by means of high-performance liquid chromatography (HPLC). To characterize the sensitivity of the method by establishing the limit of detection and limit of quantification for the following impurities: benzylamine, isonicotinic acid (raw materials for API synthesis), isonicotinic acid benzylamide (intermediate product of API synthesis) and unidentified impurities. To confirm the method compatibility with validation characteristics as recommended by the International Conference on Harmonization of Technical Requirements for the Registration of Medicinal Products for Human Use (ICH). Materials and methods. The liquid chromatograph Agilent 1200 equipped with degasser, four-channel pump, auto sampler, chromatographic column thermostat and diode array detector was used. The Zorbax Eclipse XDB-C18 chromatographic column with dimensions 150.0 ×4.6 mm filled by octadecylsilane sorbent with a particles size of 5 μm, was used. The mobile phase was a mixture of a buffer solution pH 2.5 containing sodium octane-1-sulfonate in an amount of 0.1 %: water : acetonitrile (30 : 44 : 26 v/v/v). The primary standards of Farmak JSC were used: enisamium iodide (b. 07-16) and isonicotinic acid benzylamide (b. 10413). Other materials and reagents were used with sufficient purity (Sigma-Aldrich). Results. Detection limits and limits of quantification for identified and unidentified impurities have been determined. Validation characteristics such as specificity, accuracy, precision, range of use, robustness, intermediate precision have been confirmed. The linearity has been confirmed within the range of 0.01–0.06 % for each of the impurities (with a maximum permitted content of each impurity of 0.05 %). The correlation coefficients of linear regression (r) between the entered and found values for each of the determined impurities are greater than 0.990 and the relative confidence intervals (Dz) are at the level not more than 5.0 %. Stability of the test solution, reference solution and solution for system suitability test upon storage at room temperature for 48 hours have been confirmed. Conclusions. It has been experimentally proved that the described method can be used for quality control of enisamium iodide substance.
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