Box-Behnken Design-Based Development and Validation of RP-HPLC Method for Determination of Related Substances in...

Area of Science:

• Analytical Chemistry
• Pharmaceutical Analysis

Background:

• Nitrofurantoin is an antibiotic susceptible to degradation.
• Ensuring the quality and safety of nitrofurantoin formulations requires accurate quantification of related substances and impurities.
• Existing analytical methods may not fully address the stability-indicating requirements for nitrofurantoin.

Purpose of the Study:

• To develop and validate a stable reversed-phase high-performance liquid chromatography (RP-HPLC) method for quantifying related substances in nitrofurantoin capsule formulations.
• To optimize the RP-HPLC method using Box-Behnken design for improved performance and dependability.
• To provide a robust analytical approach for routine quality control and regulatory compliance.

Main Methods:

• Development of a gradient elution RP-HPLC method using specific mobile phase compositions (formic acid/tetrahydrofuran and methanol/acetonitrile/tetrahydrofuran).
• Utilized Box-Behnken design, a response surface methodology, for systematic optimization of method parameters.
• Employed an Inertsil ODS-3 C18 column with detection at 285 nm and a column temperature of 30°C.
• Validated the method according to International Council for Harmonisation (ICH) and United States Pharmacopeia (USP) guidelines.

Main Results:

• The optimized RP-HPLC method demonstrated high accuracy (92-111%), excellent precision (≤1.0% RSD), and strong linearity (R² > 0.999).
• The Box-Behnken design effectively enhanced method performance, robustness, and dependability.
• The method proved to be stability-indicating, capable of detecting and quantifying nitrofurantoin degradation products and impurities.

Conclusions:

• The developed Box-Behnken design-based RP-HPLC method is a reliable and validated stability-indicating approach for analyzing nitrofurantoin capsule formulations.
• This method is suitable for routine quality control, ensuring product safety and regulatory compliance.
• The systematic optimization approach enhances confidence in the analytical results for nitrofurantoin impurities.