AQbD-based development of a stability-indicating UHPLC-PDA-QDa method for triptorelin in parenteral formulations

Abstract

A robust, stability-indicating analytical method for the quantification of triptorelin in suspension formulations was developed using reverse-phase ultra (high)-performance liquid chromatography (RP-UHPLC) under the Analytical Quality by Design (AQbD) framework. This systematic, risk-based approach enabled the efficient identification and optimization of critical method parameters, reducing reliance on traditional trial-and-error procedures. Key variables such as column type, temperature, gradient profile, and organic modifier composition were evaluated. Optimal chromatographic conditions were achieved using a YMC Triart C18 column (50 × 2.1 mm, 1.9 μm) at 53.8 °C. The mobile phase consisted of 10 mM ammonium formate buffer (pH 5.0) as phase A and acetonitrile containing 0.1% formic acid as phase B. A short 5 minute gradient elution at 0.48 mL min⁻¹, with UV detection at 280 nm, was applied. The method was subjected to forced degradation studies under hydrolytic (acidic and basic), oxidative, and thermal stress conditions to demonstrate its stability-indicating capability. These results support its overall suitability for routine quality control and regulatory applications in peptide drug analysis.