Ultra-Fast Analysis of Nitrosamines Using SPE-QQQ

Significance of the Topic

Nitrosamines are potent carcinogenic impurities found in pharmaceuticals and environmental samples. Regulatory bodies require sensitive high-throughput assays to monitor these compounds. Traditional LC-MS methods rely on lengthy chromatographic separations, limiting throughput and generating analytical backlogs.

Objectives and Overview of the Study

This work reproduces the US FDA RapidFire SPE-QQQ method and extends the panel by incorporating N-Nitrosoethylmethylamine (NEMA), N-Nitrosopyrrolidine (NPyR), and N-Nitrosopiperidine (NPIP). The aim is to achieve accurate quantitation of a broad nitrosamine panel in under 15 seconds per injection.

Used Methodology and Instrumentation

The analytical workflow utilizes automated online solid phase extraction (SPE) with a graphitic carbon cartridge on an Agilent RapidFire 400 High-Throughput Mass Spectrometry System, coupled to an Agilent Ultivo Triple Quadrupole LC/MS. Samples undergo a trap–wash–elute cycle optimized for <15-second throughput. Mobile phases consist of water and methanol with 0.1% formic acid. The APCI source operates in positive mode at 300 °C drying gas temperature, 6 L/min gas flow, 55 psi nebulizer pressure, and 350 °C APCI heater temperature.

Main Results and Discussion

Triplicate injections achieved approximately 12.5 seconds per sample, yielding 72 injections in under 15 minutes with negligible carryover. Precision was demonstrated by coefficients of variation between 4.5% and 9.0% across all analytes. Calibration curves spanning 1–100 ng/mL exhibited excellent linearity (R2 values of 0.997–0.999). The method proved robust, reproducible, and suitable for high-throughput screening applications.

Benefits and Practical Applications

The ultra-fast SPE-QQQ approach significantly reduces analysis time, solvent consumption, and operational costs. It is well suited for pharmaceutical quality control, environmental monitoring, and compliance testing where rapid and reliable nitrosamine quantitation is essential.

Future Trends and Potential Applications

Future developments may include expanding the analyte panel to cover additional nitrosamine species, integrating the workflow with continuous manufacturing and process analytical technology (PAT), and adopting advanced cartridge chemistries for enhanced selectivity. Automation and real-time monitoring solutions are expected to drive further improvements in throughput and data quality.

Conclusion

The Agilent RapidFire SPE-QQQ method delivers ultra-fast, reliable quantitation of nitrosamine impurities in under 15 seconds per sample. This scalable workflow aligns with regulatory guidelines and offers a powerful tool for high-throughput analytical laboratories.

Reference

1. FDA Updates and Press Announcements on Angiotensin II Receptor Blocker (ARB) Recalls. FDA, November 13, 2019.
2. Development and Validation of a RapidFire-MS/MS Method for Screening of Nitrosamine Carcinogen Impurities in ARB Drugs. FDA, July 24, 2019.
3. Information about Nitrosamine Impurities in Medications. FDA, February 3, 2020.