Forced degradation behavior of epidepride and development of a stability-indicating method based on liquid chromatography–mass spectrometry

Wei-Hsi Chen a,*, Yu-Yung Lin a, Yu Chang a, Kang-Wei Chang b,
Yi-Chih Hsia c
a Chemistry Division, Institute of Nuclear Energy Research, Longtan Township, Taoyuan County, Taiwan, ROC
b Isotope Application Division, Institute of Nuclear Energy Research, Longtan Township, Taoyuan County, Taiwan,
ROC
c Radiopharmaceuticals Production and Marketing Center, Institute of Nuclear Energy Research, Longtan Township,
Taoyuan County, Taiwan, ROC

A reversed-phase high-performance liquid chromatography (HPLC) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was applied to study the forced degradation behavior and stability of epidepride. 123I radioisotope-labeled epidepride, [(-)-N-{[(2S)-1-ethylpyrrolidin-2-yl]methyl}-5-iodo-2,3- dimethoxybenzamide] is a radiotracer with a high affinity for dopamine D2 receptors in the brain and has been used as an imaging agent for single-photon emissioncomputed tomography. HPLC studieswere performedusing 127I-epidepride (the nonradioactive compound), instead of 123I-epidepride, with an RP-18 column using a mobile phase consisting of methanol, acetonitrile, and ammonium acetate (pH 7.0, 10 mM). The eluent flow rate and the wavelength for HPLC detection were 0.5 mL/min and 210 nm, respectively. The ligand was exposed to acid (1 N HCl) and alkaline (1 N NaOH) media and was subjected to oxidative decomposition at room temperature using 3%H2O2 and to thermal decomposition at 50
C. After various reaction times (30 minutes, 1 hour, 2 hours, 8 hours, and 24 hours), the substances were investigated by HPLC and LC-MS/MS. Although no decomposition products were observed after the acidic, alkaline, and thermal treatments, >80% of the initial amount of 127I-epidepride was oxidized within 24 hours in the presence of H2O2. Only one major oxidation product with an m/z value of 435 was observed, in addition to the 127I-epidepride species (m/z 419). The product was characterized by LC-MS/MS fragmentation, and the deteriorated type and fragmentation pathways were proposed for epidepride.

Keywords: Epidepride, Forced degradation, Stress testing