Analysis of Nitrosamines - No. 566

Significance of the Topic

Nitrosamines are a class of potent carcinogens that can form in pharmaceutical products, food, and environmental samples.
Accurate, sensitive, and reliable analysis of nitrosamines is critical for regulatory compliance and public health protection.

Objectives and Study Overview

This application note presents a robust method for simultaneous determination of six common nitrosamines (NDMA, NMBA, NDEA, NEIPA, NDIPA, NDBA) using reversed-phase liquid chromatography coupled with tandem mass spectrometry.
The study aims to demonstrate effective separation, low detection limits, and high throughput using the Shimadzu Shim-pack Scepter C18-120 column on the Nexera X3 UHPLC platform and LCMS-8060 mass spectrometer.

Instrumental Methods

• Chromatography was performed on a Shim-pack Scepter C18-120 column (100 mm × 2.1 mm I.D., 1.9 µm) at 45 °C with a mobile phase of 0.05 % formic acid in water (A) and 0.05 % formic acid in methanol (B) at 0.4 mL/min and 10 µL injection volume.
• Mass spectrometric detection employed an LCMS-8060 system in positive APCI mode with MRM acquisition, probe voltage 4.0 kV, nebulizing gas flow 4.0 L/min, drying gas flow 3.0 L/min, DL temperature 150 °C, interface temperature 300 °C, block heater at 200 °C, and a hydrocarbon filter on the nitrogen line.
• Key instruments include the Nexera X3 UHPLC system and the LCMS-8060 tandem mass spectrometer.

Main Results and Discussion

The method achieved baseline separation of all six nitrosamines with retention times spanning a compact analytical window, enabling run times under 10 minutes.
Limits of detection and quantification were in the low parts-per-billion range, demonstrating excellent sensitivity.
MRM transitions provided high specificity and minimal background interference, while the Scepter column offered sharp peak shapes and robust performance over multiple injections.

Benefits and Practical Applications

• High sensitivity and selectivity for trace-level nitrosamine quantification ensure compliance with stringent regulatory limits.
• Short analysis time increases laboratory throughput for routine quality control in pharmaceutical manufacturing and food safety testing.
• Reproducible chromatography and stable mass spectrometric response support long-term method reliability and reduced maintenance.

Future Trends and Applications

Advancements may include coupling with high-resolution mass spectrometry for untargeted screening of novel nitrosamines and matrix effects studies.
Method miniaturization and faster gradients can further enhance throughput and reduce solvent consumption.
Expansion into environmental water and personal care product analysis is anticipated as regulatory requirements evolve.

Conclusion

The combination of the Shim-pack Scepter C18-120 column and the LCMS-8060 system provides a powerful platform for rapid, sensitive, and reliable nitrosamine analysis.
This workflow meets the needs of modern laboratories focused on ensuring product safety and regulatory compliance.

References

Application News 01-00188A (JP, ENG), Shimadzu Corporation First Edition Sep. 2024