HPLC Analysis of Nitrosamines on Supel™ Carbon LC
Materials
analytical column
mobile phase component
standard
N-Nitrosodiisopropylamine (NDIPA)
certified reference material, pharmaceutical secondary standardN-Nitrosoethylisopropylamine (NEIPA / NIPEA)
certified reference material, pharmaceutical secondary standardN-Nitrosomethylaminobutyric Acid (NMBA)
certified reference material, pharmaceutical secondary standardN-Nitrosomethylphenylamine (NMPA)
certified reference material, pharmaceutical secondary standardwater purification system
Milli-Q® IQ 7005 Ultrapure and Pure Water Purification System
Produces ultrapure (Type 1) water and pure (Type 2) water with a production flow rate of 5 L/hr from tap water feed.CONDITIONS
column
Supel™ Carbon LC 2.7 µm 100 x 3.0 mm I.D. (59993-U)
mobile phase
[A] Water + 0.1% TFA,[B] Acetonitrile + 0.1% TFA
gradient
0-0.5 min 2.5% B, in 4.5 min to 95% B, 2 min 95% B, 3 min at 2.5% B
flow rate
0.5 mL/min
pressure
80 – 130 bar
column temp.
90 °C
detector
UV, 230 nm
injection
5 µL
sample/matrix
six nitrosamines standard mix (~20 mg/L each) in mobile phase A (water + 0.1% TFA)
Description
General description
Nitrosamines have been known to exist in the environment and within food sources, with awareness further raised in 2018 when many regulatory authorities for medicines became aware of the dangers of N-Nitrosamines as drug contaminants. This class of compounds is known to be potent carcinogens and can be introduced through multiple introduction pathways. In this application, 2.7 µm porous graphitic carbon particles in Supel™ Carbon LC column are used to separate six different nitrosamines, followed by UV detection.
Analysis Note
The Supel™ Carbon LC column is used for the separation of six nitrosamine compounds. The column is able to separate six nitrosamines based on reversed phase with good reproducibility for repeated analyses (data not shown here). The resolution and peak asymmetry for the six different nitrosamines are greater than 1.5 and less than 2.0, respectively. The LOD and LOQ of the tested nitrosamines are in the low mg/L range and can be further improved with the use of mass spectrometry instrumentation.
Other Notes
APP_661
Legal Information
Supel is a trademark of Sigma-Aldrich Co. LLC