Development and Validation of a Customized Amplex UltraRed Assay for Sensitive Hydrogen Peroxide Detection in...

Area of Science:

• Pharmaceutical technology
• Analytical chemistry
• Cleanroom technology

Background:

• Hydrogen peroxide (H₂O₂) decontamination is crucial for cleanroom compliance.
• Detecting residual H₂O₂ in water for injection (WfI) is challenging but necessary.
• Current methods lack sensitivity for trace H₂O₂ in WfI.

Purpose of the Study:

• Develop and validate a quantitative assay for H₂O₂ in WfI.
• Achieve low nanomolar to low micromolar detection limits (ppb range).
• Ensure high accuracy and precision for H₂O₂ quantification.

Main Methods:

• Utilized the Amplex UltraRed fluorometric technology.
• Developed a customized assay kit, as commercial options were insufficient.
• Validated the assay for selectivity, sensitivity, accuracy, precision, and robustness.

Main Results:

• Successfully developed a validated quantitative fluorometric assay for H₂O₂ in WfI.
• The assay accurately quantifies H₂O₂ at ppb levels.
• Demonstrated robustness and reliability for process development.

Conclusions:

• The developed Amplex UltraRed assay enables reliable H₂O₂ detection in WfI.
• This is critical for safe and effective H₂O₂ decontamination of cleanrooms and isolators.
• The assay supports process development and qualification in pharmaceutical manufacturing.