Development and validation of high-performance liquid chromatography method for analysis of b-CCT and its related substances

Compound 2 beta-carbomethoxy-3 beta-(4-chlorophenyl)tropane (beta-CCT) is a key intermediate for the synthesis of some clinical dopamine transporter (DAT) imaging agents. Potential impurities from synthesis process of beta-CCT and degradation during storage might have detrimental effect on the final imaging agents. Thus, it is necessary to guarantee the quality of beta-CCT. In this study, a rapid, sensitive and accurate high-performance liquid chromatography (HPLC) method was devel-oped and validated for the analysis of beta-CCT and its related substances. The chromatographic sep-aration was achieved on a reverse-phase phenomenexTM Gemini C18 column with an isocratic mobile phase consisted of methanol, water and TFA (30:70:0.1 v/v/v). The flow rate was 1.0 mL/ min at 30 degrees C and samples were monitored at 220 nm. The method was validated concerning system suitability, linearity, accuracy, precision, specificity, robustness and stability. The limit of detection (LOD) and the limit of quantification (LOQ) of beta-CCT were 0.5 and 1.5 mu g/mL, respectively. The linearity range of beta-CCT was 1.5-450 mu g/mL with a good linear correlation coefficient (R-2 = 0.9999) between the peak response and concentration. Specificity investigation through forced degradation experiments displayed that beta-CCT was stable in acidic, thermal and photolytic degradation conditions, but significantly unstable in alkaline and oxidative conditions. With the developed chromatographic method, possible impurity alpha-CCT from synthetic process and potential degradation products could be well separated from beta-CCT. Good recovery and precision were manifested in the assay method. These results indicated that the present method would be suitable for not only the quality assurance of beta-CCT in regular production sample assays but also the monitoring and determination of its related substances. (c) 2022 The Author(s). Published by Elsevier B.V. on behalf of King Saud University.