QbD-based Development and Validation of RP-HPLC Method for the Estimation of Alogliptin in Human Plasma

Background: Alogliptin, developed by Takeda Pharmaceutical Company, is a DPP-4 inhibitor used in the treatment of Type II diabetes. It enhances the activity of incretin hormones by inhibiting their breakdown, which helps improve blood glucose control. It is administered orally, either alone or with other antidiabetic medications. Objective: To develop and validate a reliable and sensitive RP-HPLC method for estimating Alogliptin in human plasma, using a Quality by Design (QbD) approach for method optimization. Materials and Methods: Chromatographic separation was performed using an Agilent 1100 HPLC system and a C18 column (4.6 mm × 250 mm, 5 µm) packed with superficially porous silica particles. Plasma samples were processed through a protein precipitation technique. Method optimization was conducted using Central Composite Design (CCD), a component of QbD. Results: Optimal conditions included a mobile phase of acetonitrile and 0.1% formic acid (16.1:83.9 v/v) at a flow rate of 0.64 mL/min. Alogliptin was eluted at 5.1 minutes with peak tailing below 2. The method showed linearity in the range of 2–10 µg/mL (R² = 0.9998). It demonstrated high accuracy (99.71–101.76%) and precision, with an LOD of 0.0752 µg/mL and LOQ of 0.2297 µg/ml. Conclusion: The method was successfully developed and validated following ICH guidelines. Its reliability and sensitivity make it suitable for use in preclinical and clinical studies for estimating Alogliptin in human plasma.