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Home / Applications / Rapid HPLC Analysis of Melamine in Dairy Products

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Rapid HPLC Analysis of Melamine in Dairy Products

By Satoe Iijima

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June 14, 2024

Introduction

In this report, we will present the results of the analysis of melamine in a milk products using HPLC with a photometric diode-array (PDA) detector.

Melamine is an organic nitrogen compound that has a triazine ring at the center of its structure. It is typically used as  melamine resin, which has excellent properties of high thermal and water resistance and is used for mechanical strengthening. Recently, melamine has been found as an additive in dilute milk to make-up for the lack protein content, however, this milk product causes serious health problems. The Food and Drug Administration (FDA) has been evaluating the risks of contamination in food products  and has suggested HPLC with UV detection as one of the practical methods for the analysis of melamine.

Melamine Analysis HPLC System
LC-4000 Series HPLC System

Experimental

Sample preparation: 20 µL of 100 µg/L a melamine standard solution was added to 180 µL of milk, then diluted with 1800 µL of ultrapure water and made up to a total volume of 2mL. The resulting solution was centrifuged at 5000 rpm for 10 minutes and the supernatant filtrated using a 0.45 µm membrane filter.

 

Keywords

620020H, HPLC, PDA Detector, analysis of melamine, milk, UV detection

Results

Figure 1 shows the structure of melamine, while figure 2 shows the chromatogram and contour plot of melamine.

Molecular Structure of Melamine
Fig. 1 Molecular structure of melamine
HPLC Separation of Melamine
Fig. 2 Chromatogram of melamine

 

Fig. 3 shows the UV absorption spectrum of the melamine standard.

UV Spectrum of Melamine absorption
Fig. 3 UV absorption spectrum of a melamine standard

Figure 4 shows the chromatogram and on peak spectrum of milk spiked with melamine. The spectrum of a melamine standard registered in a search library returned a correlation coefficient of 1.0

Fig. 4 Chromatogram of milk spiked with melamine.

References

Environ Health Perspect. 2009 Dec; 117(12): 1803–1808.
Published online 2009 Aug 6. doi: 10.1289/ehp.0900949
PMCID: PMC2799451
PMID: 20049196
The Melamine Incident: Implications for International Food and Feed Safety
Céline Marie-Elise Gossner, Jørgen Schlundt,* Peter Ben Embarek, Susan Hird, Danilo Lo-Fo-Wong, Jose Javier Ocampo Beltran, Keng Ngee Teoh, and Angelika Tritscher

Access publication

This document has been prepared based on information available at the time of publication and is subject to revision without notice. Although the contents are checked with the utmost care, we do not guarantee their accuracy or completeness. JASCO Corporation assumes no responsibility or liability for any loss or damage incurred as a result of the use of any information contained in this document. Copyright and other intellectual property rights in this document remain the property of JASCO Corporation. Please do not attempt to copy, modify, redistribute, or sell etc. in whole or in part without prior written permission.

Featured Products:

  • ChromNAV 2.0

  • LC-4000 Series Preparative HPLC

  • LC-4500 Series Compact HPLC

About the Author

Ms Iijima is a member of the chromatography separations team at JASCO Corporation main applications laboratory in Tokyo Japan.

JASCO Application Note

Rapid HPLC Analysis of Melamine in Dairy Products

Introduction

In this report, we will present the results of the analysis of melamine in a milk products using HPLC with a photometric diode-array (PDA) detector.

Melamine is an organic nitrogen compound that has a triazine ring at the center of its structure. It is typically used as  melamine resin, which has excellent properties of high thermal and water resistance and is used for mechanical strengthening. Recently, melamine has been found as an additive in dilute milk to make-up for the lack protein content, however, this milk product causes serious health problems. The Food and Drug Administration (FDA) has been evaluating the risks of contamination in food products  and has suggested HPLC with UV detection as one of the practical methods for the analysis of melamine.

Melamine Analysis HPLC System
LC-4000 Series HPLC System

Experimental

Sample preparation: 20 µL of 100 µg/L a melamine standard solution was added to 180 µL of milk, then diluted with 1800 µL of ultrapure water and made up to a total volume of 2mL. The resulting solution was centrifuged at 5000 rpm for 10 minutes and the supernatant filtrated using a 0.45 µm membrane filter.

 

Results

Figure 1 shows the structure of melamine, while figure 2 shows the chromatogram and contour plot of melamine.

Molecular Structure of Melamine
Fig. 1 Molecular structure of melamine
HPLC Separation of Melamine
Fig. 2 Chromatogram of melamine

 

Fig. 3 shows the UV absorption spectrum of the melamine standard.

UV Spectrum of Melamine absorption
Fig. 3 UV absorption spectrum of a melamine standard

Figure 4 shows the chromatogram and on peak spectrum of milk spiked with melamine. The spectrum of a melamine standard registered in a search library returned a correlation coefficient of 1.0

Fig. 4 Chromatogram of milk spiked with melamine.

Keywords

620020H, HPLC, PDA Detector, analysis of melamine, milk, UV detection

References

Environ Health Perspect. 2009 Dec; 117(12): 1803–1808.
Published online 2009 Aug 6. doi: 10.1289/ehp.0900949
PMCID: PMC2799451
PMID: 20049196
The Melamine Incident: Implications for International Food and Feed Safety
Céline Marie-Elise Gossner, Jørgen Schlundt,* Peter Ben Embarek, Susan Hird, Danilo Lo-Fo-Wong, Jose Javier Ocampo Beltran, Keng Ngee Teoh, and Angelika Tritscher

Access publication

This document has been prepared based on information available at the time of publication and is subject to revision without notice. Although the contents are checked with the utmost care, we do not guarantee their accuracy or completeness. JASCO Corporation assumes no responsibility or liability for any loss or damage incurred as a result of the use of any information contained in this document. Copyright and other intellectual property rights in this document remain the property of JASCO Corporation. Please do not attempt to copy, modify, redistribute, or sell etc. in whole or in part without prior written permission.
28600 Mary’s Court, Easton, MD 21601 USA • (800) 333-5272 • Fax: (410) 822-7526 • jascoinc.com/applications

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