Using a Highly Sensitive and Selective Single-Quad LCMS-2050 for Cleaning Validation

Importance of the Topic

Effective cleaning validation prevents cross contamination in pharmaceutical production by verifying removal of trace residues of active ingredients and cleaning agents. Traditional UV based methods may lack sensitivity for compounds with poor UV absorbance. A single quadrupole LCMS system addresses this limitation by offering selective and ultra sensitive detection at picogram levels.

Objectives and Study Overview

This study evaluates coupling of a single quadrupole mass spectrometer LCMS 2050 to an UHPLC PDA system for quantitative cleaning validation. Four macrolide antibiotics were selected as test analytes to demonstrate detection limits at low nanogram and picogram levels across a wide concentration range.

Methodology and Instrumentation

Sample Preparation

• Four macrolides spiramycin azithromycin erythromycin clarithromycin
• Stock standards at 1000 microgram per mL diluted to final mixtures from 0.02 to 100000 ng per mL

Chromatography

• UHPLC system Nexera XR with PDA detector
• Reversed phase column with gradient mobile phase of acetonitrile and water with 0.1 percent formic acid
• Optimized gradient run of approximately three minutes

Mass Spectrometry

• LCMS 2050 single quadrupole with heated DUIS ion source combining ESI and APCI
• Positive ion mode scan m/z 150 to 1000 and selected ion monitoring m/z 734 748 749 843
• Interface voltage 0.5 kV corona needle 0.5 kV DL Qarray 20 kV
• Gas flows for nebulizing dry and heating gases with interface temperatures at 300 and 250 degrees C

Results and Discussion

All four macrolides produced protonated ions detected with high sensitivity. Calibration curves showed linearity with r2 above 0.999 over dynamic ranges up to 2000 fold. Limits of quantitation ranged from 0.02 to 0.5 ng per mL corresponding to 0.1 to 2.5 pg on column. Reproducibility at LOQ had relative standard deviations below nine percent and signal to noise ratios above eleven. Chromatographic overlap of spiramycin and azithromycin did not affect quantitation due to distinct m/z values. A novel software function mass it overlays MS peak information onto UV traces enabling simultaneous review of UV and MS data and preventing undetected compounds in UV analysis.

Benefits and Practical Applications

Integration of a single LCMS device with UHPLC UV delivers ultra low level detection suitable for cleaning validation where traditional methods fail. The approach combines specificity of UV separation with sensitivity of mass detection enabling quantitation of poorly absorbing compounds at picogram levels within a standard QC workflow.

Future Trends and Opportunities

Ongoing advances may include deeper integration of MS software tools for real time overlay with UV data wider adoption in quality control environments and adaptation to diverse analyte classes. The trend toward compact sensitive instruments will support in line monitoring and streamlined validation processes.

Conclusion

This study demonstrates that a single quadrupole LCMS system offers a robust solution for cleaning validation achieving limits of quantitation down to 0.02 ng per mL and accurate quantitation across four orders of magnitude. The addition of mass it functionality enhances data review by combining UV and MS perspectives within one platform.