Preparation of a standard Melamine solution

Regarding the Melamine Crisis 

Methods for Melamine Analysis via HPLC 

There are basically 2 methods we can offer via HPLC, 

depending on the requirements of the employed equipment: 

1 st Method: Reversed Phase Mode 

U.S. FDA states to use a commercially available C8 column for the analysis of Melamine. 

Very promising results can be obtained with Inertsil C8-3 

Inertsil C8-3 5um 4.6X150mm or 250mm 

(Cat# 5020-01900 or 5020-01901) 

Important Remarks on the 1 st method: Reversed Phase Mode 

An ion-pair reagent is used in this application. 

Therefore, this application is NOT compatible with LC/MS/MS analysis. 

This is due to the fact that an ion-pair reagent will heavily contaminate the expensive LC/MS detector. 

This application data is ONLY compatible with instrumental setups employing an UV or PDA detector. 

2 nd Method: For LC/MS/MS 

Please see Slide 3


1st Method: Reversed Phase Mode, Inertsil C8-3 5um 4.6X150mm


1st Method: Reversed Phase Mode, Inertsil C8-3 5um 4.6X250mm


LC/MS/MS-Method 

2 nd Method: For LC/MS/MS 

If you will be using a LC/MS/MS, please use the following: 

Inertsil HILIC 5um 3.0X150mm on LC/MS/MS-System 

(Cat# 5020-07715) 

Important Remarks on the 2 nd method: For LC/MS/MS 

The mobile phase used in this application data is compatible with LC/MS/MS.

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2 nd Method: For LC/MS/MS, Inertsil HILIC 5um 3.0X150mm

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Preparation of a standard Melamine solution 

Background information of Melamine (2, 4, 6-triamine-s-triazine) 

It is only slightly soluble in water. 

It is soluble to warm (hot) water and diluted hydrochloric acid. 

However, it is insoluble in hydrophobic solvents. 

Preparation of standard stock solution of Melamine 

1) Measure 50mg of Melamine. 

2) Dissolve it by 20% Methanol solution and making it at 50mL. 

3) Use this solution as a 1,000ppm standard stock solution. 

Preparation of standard Melamine solution 

1) Whenever using the above standard stock solution, always add 1% trichloroacetic acid and dilute it. 

2) Then use the solution as a standard Melamine solution. 

For example: 

1) Take 1mL of the standard stock solution and add 1% trichloroacetic acid and make it at 10mL. 

2) This 10mL will be the standard Melamine solution. 

3) In this case, 0.1mg of Melamine will be contained in a 1mL of standard Melamine solution.

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Procedure of Sample Preparation

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Solid Phase Extraction Method

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Ordering Information 

* For price information, please send a quotation request: 

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Preparation of a standard Melamine solution

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