A reversed-phase high-performance liquid chromatography (RP-HPLC) method has been developed using an ODSC18
column for the analysis of teriflunomide (TRF), in accordance with the regulatory requirement of Analytical Quality by Design
(AQbD) in the development of robust analytics. Following risk analysis, a number of critical method variables (CMV) were used
as inputs, including the solvent modifier (A), flow rate (B), injection volume (C), column tem (D), and buffer strength (E), while
the corresponding Flow rate (F1), injection volume (F2), and buffer strength (F3) of TRF were used as method responses that are
likely to have an impact on method performance. The analytical target profile (ATP) was established in accordance with ICH and
USP standards. The Central Composite Design (CCD) and Fractional-Factorial Design(FFD) was used as a tool in the
investigation of scientific understanding between input variables (A-E) and method responses (F1-F3). The method operable
design region (MODR) was arrived at by subsequent analysis of systematic simulation followed by 3D-Contour plots. There were
Five candidate methods were selected within MODR and verified experimentally. The r2 value was 0.997 (TRF) indicating the
consistency of the model and reliability of the method in the region. Among the methods, a method Solvent modifierACN:0.1AA(A), the Flow rate of 0.75 µM/ml (B), Injection volume of 15 µL/ml (C) Column temp of 27°C (D) and Buffer
strength of 50 µM (E) was validated as per ICH Q2R1 guidelines. A conventional RP- HPLC method was developed by trialand-error method and was compared with the AQbD method. The result indicated the AQbD method was relatively more robust
with CMV range ofACN-0.1AA (A), 0.5-1.0(B), 10-20 (C), 25-30(D)and 50-30(E) whilst the conventional HPLC method failed
for robustness.
Key words: AQbD, Teriflunomide, RP-HPLC, Method Development
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