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Table 3 Control strategy for stability methods of AV obtained by CCD design

From: Comprehensive stability-indicating method development of Avanafil Phosphodiesterase type 5 inhibitor using advanced Quality-by-Design approach

Factors Attribute or parameter Range studied Optimized data Proposed range Purpose of control
Mobile phase ratio (Buffer:ACN) Resolution 40:60, 50:50, 60:40, 70:30, 80:20 v/v 60:40, v/v 60:40 ± 2, v/v Better separation between peaks, to get > 2 resolution
pH of buffer Resolution 2.5, 3.5, 4.5, 5.5, 6.5 4.5 4.5 ± 0.2 Better separation between peaks, to get > 2 resolution
Flow rate of mobile phase Resolution 0.75, 0.5, 1, 1.25 mL/min, 0.9 mL/min 0.9 ± 0.2 mL/min Better separation between peaks, to get > 2 resolution
Temperature of column oven Resolution 0 °C (no control),
10, 20, 30, 40 °C,
20 °C 20 ± 2 °C Effective parameter for changing band spacing and improving resolution
Organic phase type Resolution ACN,
MeOH
ACN ACN To improve peak shape and resolution
Degase time Resolution 10–25 min 12–15 min (12–15 min) ±  2 To remove air bubble from mobile phase
Sample volume Resolution 5–20 μL 10 μL 10 μL ± 0.1 To reduce chances of more sample loading
Column type Resolution C18, C8 C18 C18 For better separation and resolution
Buffer type Resolution Ammonium formate,
ammonium acetate
Ammonium acetate Ammonium acetate Ammonium salts are used for LC–MS compatible method
Buffer strength Resolution 5–10 mM 10 mM 10 mM Buffer used to maintain consistent retention, reproducibility, and selectivity
Detection wavelength Resolution 230–250 nm 239 nm 239 nm More absorbance at wavelength maxima