Rapid Identification of Polysorbates 20 and 80 Directly Through Amber Bottles

Significance of the Topic

Polysorbate 20 and 80 are widely used excipients in biopharmaceutical formulations, present in approximately 80% of monoclonal antibody products. Their degradation through hydrolysis or auto‐oxidation can compromise drug stability, making reliable identification and verification of these raw materials essential for quality control and regulatory compliance.

Objectives and Study Overview

This study aimed to develop and validate a non‐invasive Raman spectroscopic method using Spatially Offset Raman Spectroscopy (SORS) for rapid differentiation of Polysorbate 20 (PS 20) and Polysorbate 80 (PS 80) through amber glass bottles. A comparison was made between conventional back‐scattering Raman and Agilent Vaya handheld Raman with SORS to assess selectivity and robustness in real‐world warehouse conditions.

Used Instrumentation

• Agilent Vaya handheld Raman spectrometer equipped with SORS technology and 830 nm laser.
• Conventional handheld Raman spectrometer with 785 nm laser for comparative analysis.

Methodology

A method development wizard built into the Vaya system generated spectral libraries for PS 20 and PS 80, using multi‐compendial grade samples in standard amber glass bottles. The system automatically applied baseline correction and container subtraction algorithms to isolate raw material spectra. Specificity tests involved ten replicate challenges per formulation, including cross‐testing each PS20 and PS80 model against both analytes. Conventional Raman measurements were acquired under the instrument’s default settings without additional processing.

Main Results and Discussion

SORS‐enabled spectra exhibited distinct Raman bands for PS 20 and PS 80, notably the ~1650 cm⁻¹ band associated with the monooleate group in PS 80. The challenge matrix for the Vaya system showed 100% correct identification of each polysorbate and <0.1 false positives off‐diagonal. In contrast, conventional Raman through amber glass yielded weak, fluorescence‐dominated spectra that failed to resolve the characteristic peaks, resulting in poor selectivity.

Benefits and Practical Applications

• Non‐destructive, through‐container testing preserves sterility and shelf‐life, reducing waste and the need for reagents.
• Rapid raw material release in warehouse environments, improving throughput and safety by minimizing sample handling.
• Elimination of sampling booths and protective clothing requirements, lowering labor and operational costs.

Future Trends and Opportunities

SORS‐based handheld Raman spectroscopy can be extended to other critical materials with low Raman scattering cross‐sections housed in light‐filtering or opaque containers. Integration with digital supply chain platforms and AI‐driven spectral libraries may further streamline raw material verification and support real‐time quality assurance workflows.

Conclusion

The Agilent Vaya Raman spectrometer with SORS technology offers a robust, selective, and rapid method for identifying Polysorbate 20 and 80 through amber glass bottles. This approach enhances raw material management in pharmaceutical settings, reduces contamination risks, and supports efficient regulatory compliance.

Reference

• ICH Q7: Good Manufacturing Practice Guide for Active Pharmaceutical Ingredients. ICH Harmonised Tripartite Guideline. Page 16. Accessed April 2021.