Track: Manufacturing and Analytical Characterization - Chemical - Analytical - Impurities and Degradation - Impurity Quantitation
Category: Poster Abstract
Chiral HPLC Method to Determine the D-/L-Epinephrine Ratio in Expired Epinephrine Autoinjectors
Purpose: Epinephrine autoinjectors (EAI) are recommended for prehospital treatment of anaphylaxis. Epinephrine is an unstable chemical. Even in the presence of sodium metabisulfite, it is susceptible to oxidation and inactivation by partial racemization to the dextroisomer. Our purpose is to evaluate unused and outdated EAIs, obtained from patients, for epinephrine content and the degradation products including the D/L enantiomer ratio. Methods: Chiral separation of epinephrine enantiomers was performed on an ORpak CDBS-453 4.6 x 150 mm column (Shodex) using an Agilent HPLC. The mobile phase was composed of 95%:5% (v/v) aqueous/acetonitrile mixture. The aqueous phase is a 200 mM sodium chloride solution containing prepared 0.05% glacial acetic acid. A flow rate of 0.5 mL/min was used in isocratic mode and the column was cooled to 10°C. The injection volume was set to 10 µL, and UV detection was monitored at 280 nm. Results: The chiral HPLC method was validated according to the ICH guidelines with calibration curves for the following standard solutions: L-(-)-Epinephrine (Fisher Scientific), D-(+)-Epinephrine (Santa Cruz Biotechnology, Inc.), and racemic DL-epinephrine (Fischer Scientific). The total chromatographic analysis time per sample was 20 minutes, and a complete baseline resolution was obtained for the D-, and L- epinephrine (Figure 1 and 2). The method was used for determination of the D/L racemic ratio of epinephrine in expired EAIs. Conclusion: Use of chiral separation of epinephrine enantiomers measured D- and L- epinephrine from expired EAIs. Knowing the amount of active and inactive epinephrine remaining in expired EAIs may allow the expiration dating to change, thus allowing patients access to EAIs for a longer duration.