(M1030-09-50) Development of an Automated and High Throughput UHPLC/MS Based Workflow for Cleaning Verification of Potent Compounds in the Pharmaceutical Manufacturing Environment

Purpose: Cleaning verification (CV) is a critical step in the pharmaceutical manufacturing process to eliminate or reduce unacceptable contamination of a product as a result of insufficiently cleaned equipment surfaces. The main concern is cross contamination with active pharmaceutical ingredients (APIs) from previous runs that may impact patient safety. Current conventional approaches involve rather tedious sample preparation and analytical methods with relative lengthy analysis time. Potent APIs possessing low acceptable daily intake (ADE) values require analytical methods for CV with very low detection limits to confirm that these APIs are below their acceptance limits prior to the next manufacturing process.

Methods: In this work, a novel end to end CV workflow was developed, which includes the automated sample and calibration solution preparation as well as high throughput analysis by ultra-high-performance liquid chromatography (UHPLC) coupled with single quadrupole mass spectrometry in flow injection analysis and selected ion monitoring mode (FIA-MS-SIM). The method was validated using ten model compounds.

Results: Acceptable specificity, linearity (R² > 0.995) and single digit ng/mL LOQ and LOD were achieved for all model compounds. This approach was also successfully applied to the analysis of 22 internal CV samples from an internal program.

Conclusion: A high-throughput, LC-FIA-MS method coupled with automated sample and calibration solution preparation was developed to quantify potent compounds during cleaning verification to support drug substance and drug product manufacturing. Our approach is based on a broadly available analytical instrumentation in the pharmaceutical industry, HPLC coupled with single quadruple MS analyzer.