Analysis of 10 Nitrosamines

Význam tématu

Nitrosamines are potent genotoxic impurities subject to strict regulatory limits across major agencies such as ICH, FDA and EMA. Their presence in pharmaceuticals and environmental samples poses significant health risks, necessitating sensitive and reliable analytical methods for routine monitoring and quality control.

Cíle a přehled studie / článku

This application note details the development of a reversed-phase LC–MS/MS method for the simultaneous determination of ten common nitrosamines. The study aims to achieve fast separation, high sensitivity, and robust quantitation using a Shim-pack Velox Biphenyl column coupled with a triple quadrupole mass spectrometer.

Použitá metodika

The chromatographic separation employs a gradient elution with mobile phases of water/formic acid (1000:0.5) and methanol/formic acid (1000:0.5). The Shim-pack Velox Biphenyl column (100 mm × 2.1 mm, 2.7 μm) operates at 45 °C with a flow rate of 0.6 mL/min. Injection volume is set to 20 μL. The gradient program runs from 1 % to 98 % organic over a 10-minute cycle, followed by re-equilibration.

Použitá instrumentace

• LC system: Shimadzu Nexera X3
• Column: Shim-pack Velox Biphenyl, P/N 227-32015-03
• Mass spectrometer: Shimadzu LCMS-8060NX with APCI in positive mode
• Ion source parameters: probe voltage 4.0 kV, nebulizing gas 4.0 L/min, drying gas 3.0 L/min, interface 350 °C, heat block 200 °C, DL 150 °C
• Detection mode: MRM
• UV detection: 254 nm (SPD-M40)

Hlavní výsledky a diskuse

The method achieved baseline separation of all ten nitrosamines—including NDMA, NDEA and NDBA—within 10 minutes. MRM transitions provided high specificity, while APCI source conditions ensured low background and robust signal intensities. The rapid gradient and biphenyl stationary phase enhanced selectivity for aromatic nitrosamine structures. Reproducibility tests demonstrated retention time RSDs below 0.5 %, supporting method robustness.

Přínosy a praktické využití metody

• Fast throughput suitable for high-volume testing in QA/QC laboratories
• High sensitivity and selectivity in a single run
• Compliance with pharmacopeial guidelines (EP, USP, JP)
• Adaptable to different nitrosamine profiles and sample matrices

Budoucí trendy a možnosti využití

• Integration with automated sample preparation workflows
• Application of high-resolution MS for non-target nitrosamine screening
• Miniaturized columns for reduced solvent consumption
• Development of green chromatography techniques

Závěr

The proposed LC-MS/MS approach on a bimodal biphenyl column delivers a rapid, sensitive and reproducible platform for nitrosamine analysis, aligning with regulatory expectations and facilitating routine pharmaceutical testing.

Reference

• Shimadzu Corporation. Application News an_01-00400. First Edition: Aug. 2023.