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Journal of Chemistry
Volume 2013, Article ID 239342, 7 pages
Research Article

A Simple HPLC-DAD Method for the Analysis of Melamine in Protein Supplements: Validation Using the Accuracy Profiles

Department of Pharmacy, University of Naples Federico II, Via D. Montesano 49, 80131 Naples, Italy

Received 7 May 2013; Accepted 15 July 2013

Academic Editor: Pranav S. Shrivastav

Copyright © 2013 Domenico Montesano et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


The study presents a fully validated simple high-performance liquid chromatography method with diode array detection (HPLC-DAD), able to accurately determine the melamine, fraudulently added, in protein supplements, commonly used from healthy adults to enhance exercise or sport performance. The validation strategy was intentionally oriented towards routine use and the reliability of the method rather than extreme performance. For this reason, validation by accuracy profile, including estimation of uncertainty, was chosen. This procedure, based on the concept of total error (bias + standard deviation), clearly showed that this method was able to determine melamine over the range of 0.05–3.0 mg Kg−1, selected by taking into account the maximum residue levels (MRLs) proposed by European legislation to distinguish between the unavoidable background presence of melamine and unacceptable adulteration. The accuracy profile procedure established that at least 95% of the future results obtained with the proposed method would be within the ±15% acceptance limits of the validated HPLC-DAD method over the whole defined concentration range.