Multi-mycotoxin occurrence in Lebanese and Italian spices and herbs

Project leaders

Michele Solfrizzo, Rajaa Fakhoury

Agreement

LIBANO - CNRS-L- not in force - National Council for Scientific Research of Lebanon

Call

CNR- CNRS (Libano) 2017-2018

Department

Biology, agriculture and food sciences

Thematic area

Biology, agriculture and food sciences

Status of the project

New

Research proposal

Spices and herbs have been used for thousands of centuries by many cultures to enhance the flavor and aroma of foods. Early cultures recognized the value of using spices and herbs in preserving foods and for their medicinal value. Scientific experiments have documented the antioxidant effects of spices and other antimicrobial properties (Rubió et al., 2013). In terms of world trade value, the most important spice crops from the tropical regions are pepper, Capsicum, nutmeg/mace, cardamom, allspice/pimento, vanilla, cloves, ginger, cinnamon and cassia, and turmeric. Coriander, cumin, mustard, and sesame seeds and the herbs sage, oregano, thyme, bay and the mints are the most important spice crops from non-tropical environments (FAO 2005). Spices, even when used in small amounts, present a potential source of microbial contamination for foodstuffs to which they are added, this is due to the environmental and processing conditions under which they are produced. Mycotoxins, especially aflatoxins and ochratoxin A, can contaminate spices and herbs at various degree of incidence and levels, therefore maximum levels (5-10 µg/kg for aflatoxins, 15-20 µg/kg for ochratoxin A) for these contaminants are in force in Europe and other Countries (Codex 2014). According to the annual report of the Rapid Alert System for Food and Feed (RASFF), in 2015, mycotoxins were the second main hazard in border rejection notifications in the European Union, being the pathogenic microorganisms the first. In the report, spices and herbs were within the food categories most affected. Most fungi present on spices and herbs are of the post-harvest and storage type, which develop after harvest if relative humidity is not controlled during storage. The most frequent toxigenic fungal contaminants of spices and herbs are species from the genera Aspergillus, Penicillium, Alternaria and less often Fusarium. Recently the Codex committee on contaminants in foods proposed a draft code of practice for the prevention and reduction of aflatoxins and ochratoxin A contamination in spices (Codex 2016). In addition to aflatoxins and ochratoxin A other mycotoxins (zearalenone, trichothecenes, fumonisins, Alternaria toxins) can also contaminate spices and herbs but their occurrence has been poorly investigated. Although the maximum permitted levels of Alternaria toxins are currently not regulated in Europe they are considered emerging mycotoxins and the introduction of maximum limits are expected in the near future. Within the Alternaria toxins, tenuazonic acid occurs in foods at the highest levels and frequencies. The U.S. Food and Drug Administration (FDA) has noted an increased number of recalls of dried spices imported from Lebanon due to mycotoxins. Spices and herbs consumed in Italy are in part produced in Italy, in part imported and in part imported and packaged in Italy. The European Committee for Standardization (CEN) has published several standard methods for the determination of mycotoxins in food commodities including spices and herbs. However, most of these methods were developed for individual mycotoxins or groups of related mycotoxins. Only aflatoxin B1 was monitored in samples of spices and herbs marketed in Italy (Romagnoli B. et al., 2007). Few studies have been done on the occurrence of mycotoxins in spices and herbs in Lebanon (Raad F., et al., 2014; Khoury A. et al., 2008). The use of LC-MS/MS technology permits to accurately determine several mycotoxins with a single method if appropriate extraction, purification and calibration approaches are used. Recently, at CNR-ISPA, 45 samples of Capsicum produced and marketed in Southern Italy have been analyzed. AFB1 was found in 30% of samples at levels up to 156 µg/kg. High levels (up to 177 µg/kg) and incidence of positive samples (52%) were found for ochratoxin A.
In addition, the presence of significant levels of fumonisins, T-2 and HT-2, zearalenone, deoxynivalenol and nivalenol were found in 59%, 22%, 91%, 35% and 37% of samples, respectively. Almost all samples were contaminated by Alternaria toxins. TeA was found in 98% of samples with a median level of 306 µg/kg. The other Alternaria toxins were also found in 9% (altenuene), 54% (alternariol), 91% (tentoxin) and 33% (alternariol mono methyl ether) of samples at level up to 1676 µg/kg. All samples were contaminated with multiple mycotoxins that range from 3 to 13 mycotoxins. The availability of two LC-MS/MS methods permitted to analyze 18 mycotoxins, the five Alternaria toxins with the first methods and the other 13 mycotoxins with the second method.
The purpose of this project is to catalogue, collect and analyze samples of the spices and herbs produced and marketed in Italy and Lebanon as well as those imported in the two countries. The samples will be spices pepper, nutmeg, cardamom, allspice, cloves, ginger, cinnamon, turmeric, coriander, cumin, mustard, and sesame seeds and herbs are sage, oregano, thyme, bay and the mints. The samples will be analysed for aflatoxins, ochratoxin A, fumonisins, T-2 and HT-2 toxins, zearalenone, deoxynivalenol and nivalenol by using an LC-MS/MS method based on the use of multi antibody column cleanup. For the determination of altenuene, alternariol, tentoxin, alternariol mono methyl ether and tenuazonic acid an LC-MS/MS method based on SPE column cleanup will be used.
The main outcomes of the project will be:
a) To identify the most frequently occurring mycotoxins in these matrices
b) To identify the spices and herbs most frequently contaminated by mycotoxins
c) To establish correlations between origin of spices and herbs and mycotoxin contamination,
d) To estimate human exposure to mycotoxins through consumption of spices and herbs in Italy and Lebanon
e) The occurrence data of mycotoxins in spices and herbs could be useful for future regulation of mycotoxins other than aflatoxins and ochratoxin A, and to set new standards for mycotoxins MRL'S.

Research goals

o To catalogue and group the spices and herbs marketed in Italy and Lebanon.
o To collect a significant and representative number of samples of spices and herbs marketed in Italy and Lebanon.
o To analyze the samples of spices and herbs for aflatoxins, ochratoxin A, fumonisins, T-2 and HT-2 toxins, zearalenone, deoxynivalenol and nivalenol by using an LC-MS/MS method based on the use of multi-antibody column cleanup.
o To analyze the samples of spices and herbs for altenuene, alternariol, tentoxin, alternariol mono methyl ether and tenuazonic acid by using an LC-MS/MS method based on solid phase extraction (SPE, OASIS HLB) column cleanup.
o To identify the most frequently occurring mycotoxins in spices and herbs marketed in Italy and Lebanon
o To identify spices and herbs most frequently contaminated by mycotoxins
o To establish correlations between the origin of spices and herbs and mycotoxin contamination
o To estimate, in Italy and Lebanon, human exposure to mycotoxins through consumption of spices and herbs
o To produce occurrence data of mycotoxins in spices and herbs that will be useful for future regulation of mycotoxins other than aflatoxins and ochratoxin A, in order to support LIBNOR in setting a standard for MRL's for spices and herbs.

Last update: 19/04/2024