Abstract
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.
Highlights
Melamine (2,4,6-triamino-1,3,5-triazine) is commonly used as a raw material or additive in the manufacture of melamineformaldehyde plastics or can coatings for food contact materials
Validation by accuracy profile, including estimation of uncertainty, was chosen. This procedure, based on the concept of total error, 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 high-performance liquid chromatography (HPLC)-DAD method over the whole defined concentration range
Summary
Melamine (2,4,6-triamino-1,3,5-triazine) is commonly used as a raw material or additive in the manufacture of melamineformaldehyde plastics or can coatings for food contact materials. It can be found at low levels as a contaminant in a variety of foods due to the leaching from food contact materials [1,2,3,4]. It is fraudulently added to food and/or feed ingredients to boost their total protein content and increase their market value Such adulteration exploits a limitation of the most common nonspecific protein tests such as Kjeldahl method, which use total nitrogen as an indicator of protein content (melamine contains 66.6% of nitrogen by weight). In order to harmonize maximum residue levels (MRLs), set in many countries all over the world, European Food Safety Authority has proposed a legal maximum residue level (MRL) of 1.0 mg/Kg for infant foods and 2.5 mg/Kg for other foods to distinguish between the unavoidable background presence of melamine (from food contact materials, pesticide use, etc.) and unacceptable adulteration [18]
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