engleski

Application of LC MS in characterization of degradation products in drug substances

The International Conference on Harmonization (ICH) guideline entitled “Stability testing of new drug substances and products” requires that stress testing has to be carried out to elucidate the inherent stability characteristics of the active substance and to identify degradation products 1. Stress testing should include the effect of temperature and humidity ; the susceptibility of drug to hydrolysis and oxidation ; and photostability. In this work the force degradation of bivalirudin, and the determination of bivalirudin in the presence of its degradation products is presented. Bivalirudin is a synthetic oligopeptide, consists of 20 amino acids (D-Phe- Pro-Arg-Pro-Gly-Gly-Gly-Gly-Asn-Gly-Asp-Phe-Glu- Glu-Ile-Pro-Glu-Glu-Tyr-Leu). Bivalirudin specifically binds to the thrombin at two sites, catalytic site and anion binding site. As a direct thrombin inhibitor, bivaliruidin is used as anticoagulant for patients with acute coronary syndromes or patients undergoing percutaneous coronary intervention 2. An UPLC method for determination of bivalirudin in the presence of its degradation products has been developed using quality-by-design approach. Developed UPLC method was validated as well, and was proven to be an effective method for separating bivalirudin from its degradation products. Forced degradation studies were performed using acid, base, H2O2, heat and light exposure as recommended by the International Conference on Harmonization (ICH). High resolution MS and MS/MS analysis were performed to propose the structure of degradation products generated under force degradation conditions. 2D-UPLC instrument was used to remove non-volatile buffer used as a mobile phase. Molecular structure was proposed for 14 degradation impurities.

bivalirudin, degradation products, LC-MS analysis

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