HPLC
IndustriesPharma & Biopharma
ManufacturerWaters
Significance of the Topic
Accurate and efficient analysis of budesonide nasal spray is essential for quality control in pharmaceutical manufacturing and regulatory compliance. Modernizing compendial HPLC monographs to UPLC methods can dramatically shorten run times, reduce solvent use, and improve laboratory throughput without compromising analytical performance.
Objectives and Study Overview
This study demonstrates the transfer of the USP monograph method for budesonide from a traditional HPLC platform to a sub-2 µm UPLC system. Goals included maintaining system suitability criteria, achieving significant reductions in run time and solvent/sample consumption, and evaluating method robustness through extended routine use injections.
Instrumentation used
- Alliance HPLC system with 2489 UV/Visible detector
- ACQUITY UPLC system with TUV detector
- XBridge C18 column, 4.6×150 mm, 5 µm (L1)
- ACQUITY UPLC BEH C18 column, 2.1×50 mm, 1.7 µm (L1)
- Waters pre-column filter and method transfer calculator tools
- Empower 2 CDS software
Methodology and Sample Preparation
The HPLC method used isocratic separation at pH 3.2 (phosphate buffer/acetonitrile, 68:32) at 1.5 mL/min over 32 minutes. Sample prep involved diluting Rhinocort AQUA nasal spray in acetonitrile and buffer, mechanical shaking, and centrifugation to yield a 12.8 µg/mL analyte solution. UPLC conditions were scaled via the ACQUITY UPLC Columns Calculator to an isocratic method (0.92 mL/min, same mobile phase) yielding a 4-minute run time while preserving retention factors.
Main Results and Discussion
System suitability criteria (resolution ≥1.5, plate count ≥5500, relative retention time 1.1) were met for both HPLC and UPLC methods. Initial routine use on UPLC showed plate count decline after ~830 injections. Method refinements—extended centrifugation (25 min), addition of a pre-column filter, and a constant column temperature (30 °C)—restored robustness. Over 2846 injections, resolution and plate count remained within USP limits and system pressure stabilized around 11,800 psi.
Benefits and Practical Applications
- 87% reduction in chromatographic run time
- Over 90% reduction in solvent and sample consumption
- Robust performance for routine QC testing over thousands of injections
- Seamless adoption of UPLC technology for compendial monographs
Future Trends and Potential Applications
Further expansion of method translation tools can facilitate rapid modernization of other USP and global pharmacopeial methods. Advances in column chemistries and AI-driven optimization may yield even shorter run times and enhanced selectivity. Integrating UPLC with high-resolution mass spectrometry could broaden its use in impurity profiling and formulation development.
Conclusion
Transferring the budesonide USP HPLC method to UPLC has been successful, delivering dramatic gains in speed, solvent economy, and routine operation robustness. The use of a method-translation calculator, optimized sample preparation, and preventive maintenance measures enabled a long column lifetime and reliable performance for over 2800 injections, demonstrating UPLC’s value for compendial analysis.
References
- USP Monograph. Budesonide, USP34-NF29, The United States Pharmacopeial Convention, official from May 1, 2011.
- USP Chapter 621. Liquid Chromatography, USP34–NF29, Pharmacopeial Forum 35(6):1516.
- Jones MD, Alden P, Fountain KJ, Aubin A. Implementation of Methods Translation between Liquid Chromatography Instrumentation. Waters Application Note, 2010;720003721EN.
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