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Simple ANP Method for Determinations of Cyanuric Acid and Melamine

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Notes: Melamine is not a food product but was found in the past in pet food and also in infant food formulas. It was alleged that melamine was added intentionally to protein products to gain protein content (test is done for nitrogen content). Melamine itself has low or no toxicity however melamine production byproducts often contain cyanuric acid, which together with melamine causes toxicity (forming insoluble crystals in the kidneys). Due to the adverse effects of melamine/Cyanuric acid contamination imported products for infants have to be tested for the presence of these potentially harmful compounds.

Method Conditions

Cogent Diamond Hydride™, 4µm, 100A

Catalog No.

A: DI water + 0.1% acetic acid
B: Acetonitrile + 0.1% acetic acid

Time	%B
0.0	100
15.00	50

0.4 mL/min. t0 = 0.8 min.

Sample Prep

1. Cyanuric acid 1.5 microg/mL: NEG (M-H)- 128.0102 m/z analyzed
in Neg mode, RT = 1.278 min
2. Melamine 3 microg/mL: POS (M+H)+ 127.0727 m/z analyzed in Pos
mode, RT = 11.260 min

ESI – NEG for cyanuric acid and ESI – POS for melamine: -
Agilent 6210 MSD TOF mass spectrometer.
Note: When an instrument with a dual ion source is used both samples can
be analyzed simultaneously.

The benefits of using the method in this application note include: 1. Short equilibration time between gradient runs 2. Excellent repeteability of the results. 3. Use of LC-MS detection eliminates the need for derivatization (as is needed with a GCMS method), 4. Higher flow rates without compromising the efficiency (higher sample throughput possible) 5. Columns used in this method have very long life and are good for over 2000 injections of real life samples. When analyzing melamine alone, a simple isocratic method can be used. When using the Cogent Diamond Hydride™ column melamine is retained at 80% B mobile phase. The resulting peak shape is very symmetrical.