Pharmaceutical Analysis Using UV-Vis: Compliance with USP Chapter <857>, and European Pharmacopoeia (Ph. Eur. Chapter 2.2.25)

Significance of the topic

UV-Vis spectroscopy is a cornerstone technique in pharmaceutical quality assurance and research. Compliance with pharmacopeial standards such as USP <857> and Ph. Eur. 2.2.25 is mandatory to ensure instrument performance and data integrity in regulated environments. Automated system verification streamlines routine checks, reduces human error, and supports GMP and 21 CFR Part 11 compliance.

Objectives and overview

This application note demonstrates how the Agilent Cary 3500 UV-Vis spectrophotometer and its Cary UV Workstation software execute system suitability tests aligned with USP and European Pharmacopoeia requirements. The goals are to:

• Review the set of automated verification tests for wavelength, photometric, stray light, linearity and resolution.
• Showcase instrument design features that facilitate simultaneous sample analysis.
• Confirm compliance with pharmacopeial tolerances across the UV-Vis range.

Methodology and instrumentation used

The Cary 3500 comprises a separation of the optical engine and a multicell module supporting eight cuvette positions without moving parts. Instrument performance tests implemented in Cary UV Workstation include:

• Control of wavelength accuracy and precision using certified reference materials: holmium oxide (200–780 nm), cerium sulfate (200–300 nm), didymium and xenon emission lines.
• Control of photometric absorbance accuracy, precision and linearity with potassium dichromate solutions (UV region) and NIST-traceable glass filters (VIS region).
• Stray light determination at cutoff wavelengths using aqueous KCl, NaI, NaNO₂ and acetone standards.
• Resolution assessment via toluene in hexane scanning between 275 and 265 nm to calculate the Aₘₐₓ/Aₘᵢₙ ratio.

Main results and discussion

Automated tests on the Cary 3500 yielded pass results within pharmacopeial limits:

• Wavelength accuracy: ±1 nm (200–400 nm) and ±2 nm (400–780 nm) with <0.5 nm precision.
• Photometric accuracy: ≤±0.01 Abs below 1 Abs and ≤±1 % above 1 Abs; photometric linearity R²≥0.999 across multiple concentrations.
• Stray light: Absorbance >2.0 A (198, 220 nm) and >3.0 A (250–370 nm) as specified.
• Resolution: A₂₆₉/A₂₆₆ ratio meeting the required spectral bandwidth performance.

These results confirm the Cary 3500’s robust optics and reliable electronics across all cuvette channels.

Benefits and practical applications

Key advantages include:

• Simultaneous measurement across eight positions, boosting throughput and internal reference control.
• Automated compliance protocols reduce manual intervention and documentation burden.
• Separation of engine and sample module simplifies qualification strategies.
• Integrated software supports data integrity, audit trails and electronic signatures.

Future trends and potential applications

Emerging developments may include:

• Enhanced connectivity with LIMS and cloud platforms for real-time performance tracking.
• Advanced chemometric algorithms for multi-component analysis.
• Miniaturized or flow-through multicell designs for process analytical technology (PAT).
• Integration with AI-driven diagnostics for predictive maintenance.

Conclusion

The Agilent Cary 3500 UV-Vis spectrophotometer, coupled with automated Cary UV Workstation verification tests, robustly meets USP <857> and Ph. Eur. 2.2.25 requirements. Its unique multicell design, comprehensive performance checks and software compliance features make it an ideal choice for pharmaceutical laboratories striving for high data quality, regulatory adherence and operational efficiency.