Determination of eight isomers and related substance of Aprepitant using normal-phase and reverse-phase HPLC methods with mass spectrophotometric detection

摘要

Introduction: Aprepitant is a chiral prodrug possessing three stereogenic centers and exists in eightstereochemical forms. It was aimed to estimate and validate Aprepitant using reverse phase HPLC.Materials and methods: Chiral liquid chromatography with an ion-trap mass spectrophotometer wasused to isolate all the isomers. The separation of the eight isomers was achieved on an amylase-basedchiral column (Chiralpak ADH, 250 mm 4.6 mm) using n-hexane/isopropyl alcohol/methanol/trifluoroaceticacid (970/40/4/0.5, vol/vol/vol/vol) as the mobile phase at a flow rate of 0.5 ml/min. Areverse-phase HPLC method was used to analyze Aprepitant and its related substances. The determinationof Aprepitant and its related substances was developed by using Xterra RP e 18,250 mm 4.6 mm, 5 mm column.Results and conclusion: These methods were validated according to the International Conference onHarmonization (ICH) guidelines. The limit of detection (LOD) and limit of quantitation (LOQ) for Aprepitantwere found to be 0.14 mg/ml and 0.41 mg/ml, respectively, for chiral substance and 0.07 mg/ml and0.21 mg/ml, respectively, for the related substance.