Ana Vulić

Abstracts

3 The Development of Liquid Chromatography Tandem Mass Spectrometry Method for Citrinin Determination in Dry-Fermented Meat Products

Authors: Jelka Pleadin, Tina Lešić, Ana Vulić, Nina Kudumija, Manuela Zadravec, Nada Vahcic, Maja Kis, Tomaz Polak

Abstract:

Mycotoxins are toxic secondary metabolites produced by numerous types of molds. They can contaminate both food and feed so that they represent a serious public health concern. Production of dry-fermented meat products involves ripening, during which molds can overgrow the product surface, produce mycotoxins, and consequently contaminate the final product. Citrinin is a mycotoxin produced mainly by the Penicillium citrinum. Data on citrinin occurrence in both food and feed are limited. Therefore, there is a need for research on citrinin occurrence in these types of meat products. The LC-MS/MS method for citrinin determination was developed and validated. Sample preparation was performed using immunoaffinity columns, which resulted in clean sample extracts. Method validation included the determination of the limit of detection (LOD), the limit of quantification (LOQ), recovery, linearity, and matrix effect in accordance to the latest validation guidance. The determined LOD and LOQ were 0.60 µg/kg and 1.98 µg/kg, respectively, showing a good method sensitivity. The method was tested for its linearity in the calibration range of 1 µg/L to 10 µg/L. The recovery was 100.9 %, while the matrix effect was 0.7 %. This method was employed in the analysis of 47 samples of dry-fermented sausages collected from local households. Citrinin wasn’t detected in any of these samples, probably because of the short ripening period of the tested sausages that takes three months tops. The developed method shall be used to test other types of traditional dry-cured products, such as prosciuttos, whose surface is usually more heavily overgrown by surface molds due to the longer ripening period.

Keywords: Mycotoxins, LC-MS/MS, citrinin, dry-fermented meat products

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2 Ochratoxin-A in Traditional Meat Products from Croatian Households

Authors: Jelka Pleadin, Tina Lešić, Ana Vulić, Nina Kudumija, Manuela Zadravec, Mario Skrivanko, Irena Perkovic, Nada Vahcic

Abstract:

Products of animal origin, such as meat and meat products, can contribute to human mycotoxins’ intake coming as a result of either indirect transfer from farm animals exposed to naturally contaminated grains and feed (carry-over effects) or direct contamination with moulds or naturally contaminated spice mixtures used in meat production. Ochratoxin A (OTA) is mycotoxin considered to be of the outermost importance from the public health standpoint in connection with meat products. The aim of this study was to investigate the occurrence of OTA in different traditional meat products circulating on Croatian markets during 2018, produced by a large number of households situated in eastern and north Croatian regions using a variety of technologies. Concentrations of OTA were determined in traditional meat products (n = 70), including dry fermented sausages (Slavonian kulen, Slavonian sausage, Istrian sausage and domestic sausage; n = 28), dry-cured meat products (pancetta, pork rack and ham; n = 22) and cooked sausages (liver sausages, black pudding sausages and pate; n = 20). OTA was analyzed by use of quantitative screening immunoassay method (ELISA) and confirmed for positive samples (higher than the limit of detection) by liquid chromatography tandem mass spectrometry (LC-MS/MS) method. Whereas the bacon samples contaminated with OTA were not found, its level in dry fermented sausages ranged from 0.22 to 2.17 µg/kg and in dry-cured meat products from 0.47 to 5.35 µg/kg, with in total 9% of positive samples. Besides possible primary contamination of these products arising due to improper manufacturing or/and storage conditions, observed OTA contamination could also be the consequence of secondary contamination that comes as a result of contaminated feed the animals were fed on. OTA levels obtained in cooked sausages ranged from 0.32 to 4.12 µg/kg (5% of positives) and could probably be linked to the contaminated raw materials (liver, kidney and spices) used in the sausages production. The results showed an occasional OTA contamination of traditional meat products, pointing that to avoid such contamination on households these products should be produced and processed under standardized and well-controlled conditions. Further investigations should be performed in order to identify mycotoxin-producing moulds on the surface of the products and to define preventative measures that can reduce the contamination of traditional meat products during their production on households and period of storage.

Keywords: Croatian households, ochratoxin-A, traditional cooked sausages, traditional dry-cured meat products

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1 Comparison of Fatty Acids Composition of Three Commercial Fish Species Farmed in the Adriatic Sea

Authors: Jelka Pleadin, Greta Krešić, Tina Lešić, Ana Vulić, Renata Barić, Tanja Bogdanović, Dražen Oraić, Ana Legac, Snježana Zrnčić

Abstract:

Fish has been acknowledged as an integral component of a well-balanced diet, providing a healthy source of energy, high-quality proteins, vitamins, essential minerals and, especially, n-3 long-chain polyunsaturated fatty acids (n-3 LC PUFA), mainly eicosapentaenoic acid (20:5 n-3 EPA), and docosahexaenoicacid, (22:6 n-3 DHA), whose pleiotropic effects in terms of health promotion and disease prevention have been increasingly recognised. In this study, the fatty acids composition of three commercially important farmed fish species: sea bream (Sparus aurata), sea bass (Dicentrarchus labrax) and dentex (Dentex dentex) was investigated. In total, 60 fish samples were retrieved during 2015 (n = 30) and 2016 (n = 30) from different locations in the Adriatic Sea. Methyl esters of fatty acids were analysed using gas chromatography (GC) with flame ionization detection (FID). The results show that the most represented fatty acid in all three analysed species is oleic acid (C18:1n-9, OA), followed by linoleic acid (C18:2n-6, LA) and palmitic acid (C16:0, PA). Dentex was shown to have two to four times higher eicosapentaenoic (EPA) and docosahexaenoic (DHA) acid content as compared to sea bream and sea bass. The recommended n-6/n-3 ratio was determined in all fish species but obtained results pointed to statistically significant differences (p < 0.05) in fatty acid composition among the analysed fish species and their potential as a dietary source of valuable fatty acids. Sea bass and sea bream had a significantly higher proportion of n-6 fatty acids, while dentex had a significantly higher proportion of n-3 (C18:4n-3, C20:4n-3, EPA, DHA) fatty acids. A higher hypocholesterolaemic and hypercholesterolaemic fatty acids (HH) ratio was determined for sea bass and sea bream, which comes as the consequence of a lower share of SFA determined in these two species in comparison to dentex. Since the analysed fish species vary in their fatty acids composition consumption of diverse fish species would be advisable. Based on the established lipid quality indicators, dentex, a fish species underutilised by the aquaculture, seems to be a highly recommendable and important source of fatty acids recommended to be included into the human diet.

Keywords: Fatty Acids, farmed fish, sea bass, dentex, sea bream

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