Simultaneous HPLC Analysis of Cyanuric Acid and Melamine on Amaze TH Mixed-Mode Column

Application description

Melamine is hydrophilic basic compound with pKa of 5. Melamine due to high nitrogen content was used as adulterant of milk products to artificially increase nitrogen content. Cyanuric acid is a trimer of cyanic acid. It is polar and slightly acidic in nature. Melamine and cyanuric acid can form a salt which cases serious health issues when consumed. Melamine and cyanuric acid carry an opposite to each other charge. We have developed a method for simultaneous analysis of melamine and cyanuric acid on Amaze TH HILIC mixed-mode column. Melamine is retained by combination of HILIC and cation-exchange mechanism. Cyanuric acid is retained by HILIC and anion-exchange mechanism. Amount of ACn, buffer pH and buffer concentration can be used to adjust retention time and resolution of both acidic and basic compounds. Method is compatible with major detection techniques 9UV, ELSD, CAD, RI, MS) and can be used for quantitation of melamine and cyanuric acid in food compositions and biofluids.Tri-modal HPLC columns provide additional flexibility in range of mobile phase which can be used to adjust selectivity of separation.

Conditions of Experiment
Column: Amaze TH
Separation Modes: HILIC, anion-exchange
Column Dimenstions: 4.6 x 150 mm, 5 um, 100A,
Mobile Phase: 85% ACN with 15 mM AmFm pH 3.8
Detection: ELSD
Sample: 0.3 mg/ml
Injection: 5 uL
Flow rate: 1 ml/min
Class of compounds: Aromatic acid, Aromatic base, Organic acid
Nature of compounds: Acidic, Basic, Hydrophilic, Polar
Compounds: Cyanuric acid, Melamine