Methodical approach for determination of the heterocyclic aromatic amines in meat products using HPLC–MS/MS

D A Utyanov,A A Kurzova,A N Ivankin,A S Knyazeva,A V Kulikovskii

doi:10.21323/2414-438x-2021-6-2-118-127

Abstract

Heterocyclic aromatic amines (HAA) are formed in foods of animal origin during the Maillard reaction due to the high creatine and creatinine contents. HAA have carcinogenic and mutagenic effects. HAA content is not standardized in the Russian Federation and the Customs Union territory. However, in the EU countries, comprehensive monitoring studies are carried out on the HAA contents and effect on the human body. Due to constant expansion of the list of controlled contaminants in food products, analytical laboratories need to develop methods for determining HAA in food items. As a result of the research, a method for HAA determination was developed using high-performance liquid chromatography with mass spectrometry in the mode of specified reaction monitoring. Comparative tests of the two methods for sample preparation were carried out. The advantages and disadvantages of sample preparation approaches were substantiated. The existing SPE conditions were optimized, which made it possible to concentrate trace amounts of MeIQx and PhIP and to dispose of substances suppressing analyte ionization. The estimation of method accuracy and specificity was carried out. The degree of ionization suppression by the matrix for MeIQx and PhIP analytes was determined. The degree of HAA extraction was empirically established. For biological samples of animal origin, it was up to 90.9% for MeIQx and up to 89.4% for PhIP. It is shown that, in accordance with the developed methodology, HAA may be determined with an accuracy of 96.15 to 98.4% at the levels of 5 to 20 ng/g. The limit of quantification of the target substances was 3 ng/g.

Highlights

Heat treatment of food products leads to the formation and/or accumulation of new compounds
The results showed that replacing 40% of animal fat in a meat product with vegetable oil led to a significant decrease in the amount of heterocyclic aromatic amines (HAA) formed during heat treatment
The purpose of this work was to substantiate the conditions for the chromatographic determination of HAA in meat products using HPLC–MS/MS and to compare the slightly modified method of sample preparation with solid-phase extraction (SPE) used in foreign laboratories and the method of sample preparation with liquid extraction by an organic solvent developed by the authors of the work

Summary

Introduction

Heat treatment of food products leads to the formation and/or accumulation of new compounds. Often, such compounds are not useful and, may harm human health due to their mutagenic and carcinogenic properties. These compounds include heterocyclic aromatic amines (HAA), which are formed in foods as a result of high-temperature treatment. According to [2], the HAA formation reaction proceeds by cleavage of water from creatine and its further cyclization to creatinine, which forms the amino-imidazole part of the HAA molecule being its basis. Pyrazines and pyridines formed from unbound amino acids and hexoses during the Maillard reaction complete the HAA molecule formation. An important participant in the HAA molecule formation is the Strecker aldehyde or the corresponding Schiff base (Figure 1)

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