REAL-TIME MONITORING OF IN-PROCESS SAMPLES BY UPLC

Importance of the Topic

Real-time monitoring of manufacturing processes is essential in pharmaceutical production to ensure consistent product quality, maximize yields, and reduce time-consuming offline analyses. Integrating high-performance liquid chromatography directly into the production environment addresses limitations of traditional spectroscopic sensors by providing selective and sensitive quantification of multiple components in complex mixtures.

Objectives and Study Overview

This work demonstrates the implementation of the PATROL UPLC Process Analyzer for both on-line and at-line monitoring of reaction streams and purification effluents. Key objectives include validating the system’s precision and speed, showcasing its ability to quantify active pharmaceutical ingredients (APIs) and impurities in real time, and evaluating its integration with process control systems.

Methodology

Two applications were investigated:

• Simulated purification effluent was generated using a quaternary gradient pump. Samples were analyzed on a 2.1×50 mm BEH C18, 1.7 μm column at 1.0 mL/min and 50 °C. A 1 min run time with a 2.67 min cycle enabled monitoring of three impurities and the API at 243 nm.
• Reaction monitoring of acetylsalicylic acid conversion to salicylic acid was performed in a heated vessel at 75 °C. Sampling was automated, and analysis used a 2.1×50 mm HSS T3, 1.8 μm column with a 2 min gradient (5–80% B), 0.8 mL/min flow, 2.5 min run time, and 4.17 min cycle.

Used Instrumentation

• PATROL UPLC Process Analyzer with integrated sample management module.
• Columns: BEH C18 (2.1×50 mm, 1.7 μm) and HSS T3 (2.1×50 mm, 1.8 μm).
• Quaternary gradient pump for effluent simulation.
• UV detector at 243 nm, 40 Hz; time constant 0.025 s.
• On-Line Manager (OLM) and At-Line Manager (ALM) software for seamless automation and barcode-based walk-up operation.

Key Results and Discussion

The system achieved high precision, with RSD values below 0.2% for peak area and height in both scenarios. On-line effluent monitoring accurately replicated UV/Vis profiles and quantified individual impurity peaks, enabling determination of the optimal collection window for maximum purity. Reaction monitoring provided real-time concentration profiles of starting material, API, and four process impurities, allowing precise identification of the reaction endpoint and quench timing. These capabilities surpass those of conventional spectroscopic sensors by resolving and quantifying low-level components in complex matrices.

Benefits and Practical Applications

• Fully automated integration with distributed control systems (DCS) and laboratory information management systems (LIMS).
• Walk-up at-line analysis with barcode scanning to reduce operator input and errors.
• Simultaneous quantification of multiple analytes, improving process understanding and control.
• Minimized offline sampling and faster feedback loops for decision-making in quality assurance and control.

Future Trends and Potential Applications

Further developments may include combining real-time UPLC data with advanced chemometric algorithms for predictive process control, miniaturizing UPLC modules for decentralized monitoring, and integrating with Industry 4.0 platforms to enable self-optimizing manufacturing lines. Expansion into bioprocessing and continuous manufacturing environments represents additional growth areas.

Conclusion

The PATROL UPLC Process Analyzer effectively brings high-performance chromatography onto the manufacturing floor, delivering precise, real-time quantification of APIs and impurities. This technology enhances process analytical capabilities, accelerates decision-making, and supports robust strategies for improving product yield and purity in pharmaceutical manufacturing.