Forced Degradation Studies of Norepinephrine and Epinephrine from dental anesthetics: Development of Stability‐Indicating HPLC Method and In Silico Toxicity evaluation

Injectable solutions containing Epinephrine (EPI) and Norepinephrine (NE) are not stable and their degradation is favored mainly by the oxidation of catechol moiety. Since studies of these drugs under forced degradation conditions are scarce, herein we report the identification of their degradation products (DP) in anesthetic formulations by the development of stability-indicating HPLC method. Finally, the risk assessment of the major degradation products was evaluated using in silico toxicity approach. HPLC method was developed to obtain a higher selectivity allowing adequate elution for both drugs and their DPs. The optimized conditions were developed by using a C18 HPLC column, sodium 1-octanesulfonate and methanol (80:20 v/v) as mobile phase, with a flow rate of 1.5 mL/min, UV detection at 199 nm. The analysis of standard solutions with these modifications resulted in greater retention time for EPI and NE, which allow the separation of these drugs from their respective DPs. Then, Five DPs were identified and analyzed by in silico studies. Most of the DPs showed important alerts as hepatotoxicity and mutagenicity. To the best of our acknowledgment, this is the first report of a stability-indicating HPLC method which can be used with formulations containing catecholamines.