Development and application of HILIC MS/MS method for determination of acrylamide in selected food samples with human health risk assessment

[S. Đekić]

120 str.

Hemija - Analitička hemija / Chemistry - Analytical Chemistry

Naučni cilj ove doktorske disertacije bio je razvoj i primena HILIC-MS/MS metode za određivanje akrilamida u odabranim uzorcima hrane sa procenom rizika po zdravlje ljudi usled izloženosti akrilamidu, pri čemu su razmatrani nekarcinogeni, karcinogeni i mutageni rizici. Razvijena je i validirana brza, jednostavna, osetljiva i robustna metoda za određivanje akrilamida u različitim prehrambenim proizvodima primenom tečne hromatografije zasnovane na hidrofilnim interakcijama spregnute sa tandemnom masenom spektrometrijom. Za pouzdanu validaciju metode pripremljen je kontrolni uzorak hleba sa dodatkom enzima asparaginaze, čime je prevaziđen nedostatak matriksa bez akrilamida. Validirani postupak pokazao je dobre analitičke performanse sa niskim granicama detekcije i kvantifikacije uz zadovoljavajući rikaveri i preciznost metode. Metoda je primenjena za određivanje sadržaja akrilamida u uzorcima sa srpskog tržišta: različitim vrstama hleba, odabranim finim pekarskim proizvodima, čipsevima i dečijoj hrani na bazi cerealija. U dečijoj hrani na bazi cerealija sadržaj akrilamida je bio ispod granice detekcije. Utvrđeno je da većina ispitanih uzoraka hleba ima koncentracije akrilamida ispod regulatornih graničnih vrednosti. Najviše koncentracije akrilamida detektovane su u integralnom pšeničnom hlebu sa semenkama, a najniže u ražanom hlebu. Ispitivanje distribucije akrilamida u hlebu pokazalo je značajno veće koncentracije u kori nego u sredini hleba. Eksperimentalno je potvrđena direktna zavisnost sadržaja akrilamida od vremena tostiranja, pri čemu je utvrđeno da tostiranje duže od 5 minuta rezultira značajnim povećanjem mutagenog rizika. U čipsu su izmerene najviše koncentracije akrilamida među svim ispitanim proizvodima. Procena rizika po ljudsko zdravlje pokazala je da ne postoji nekarcinogeni rizik, ali je utvrđen potencijalni mutageni rizik za sve ispitane uzorke, sa posebno visokim vrednostima za čips i tostirani hleb. Sa aspekta karcinogenog rizika, samo određene vrste hleba, neki uzorci biskvita i grisina pokazali su zadovoljavajući nivo bezbednosti. Ovo istraživanje predstavlja prvu procenu rizika po zdravlje stanovnika Srbije usled izloženosti akrilamidu putem hrane i pruža značajnu naučnu osnovu za razvoj strategija za smanjenje izloženosti ovom kontaminantu.

The scientific objective of this doctoral dissertation was to develop and apply a HILIC-MS/MS method for the determination of acrylamide in selected food samples, along with an assessment of human health risks associated with acrylamide exposure, considering non-carcinogenic, carcinogenic, and mutagenic risks. A rapid, simple, sensitive, and robust method was developed and validated for the quantification of acrylamide in various food products using hydrophilic interaction liquid chromatography coupled with tandem mass spectrometry. To enable reliable method validation, a control bread sample was prepared using the asparaginase enzyme, thereby overcoming the lack of an acrylamide-free matrix. The validated method demonstrated good analytical performance, with low limits of detection and quantification, and satisfactory recovery and precision. The method was applied to determine acrylamide content in samples from the Serbian market, including various types of bread, selected fine bakery products, potato chips, and cereal-based baby food. In cereal-based baby food, acrylamide was found to be below the detection limit. Most analyzed bread samples contained acrylamide concentrations below regulatory threshold values. The highest acrylamide levels were detected in wholegrain wheat bread with seeds, while the lowest were found in rye bread. Distribution analysis revealed significantly higher acrylamide concentrations in bread crust compared to crumb. A direct relationship between toasting time and acrylamide formation was experimentally confirmed, with toasting longer than 5 minutes resulting in a substantial increase in mutagenic risk. Potato chips had the highest acrylamide concentrations among all tested products. The human health risk assessment indicated no non-carcinogenic risk but identified a potential mutagenic risk for all analyzed samples, with particularly high values for chips and extensively toasted bread. From the perspective of carcinogenic risk, only certain types of bread, as well as some biscuit and snacks samples, demonstrated an acceptable level of safety. This study presents the first health risk assessment for the Serbian population related to dietary acrylamide exposure and provides a valuable scientific basis for the development of strategies aimed at reducing exposure to this contaminant.

Akrilamid, QuEChERS, HILIC-MS/MS, asparaginaza, tostiranje hleba, kora i sredina hleba, dečija hrana, fini pekarski proizvodi, čips, procena rizika

Acrylamide, QuEChERS, HILIC-MS/MS, Asparaginase, Toasting Bread, Bread Crust and Crumb, Baby Food, Fine Bakery Products, Chips, Risk Assessment

543.4:614.31(043.3)

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