Food Safety - Mycotoxins in Foods
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Transcript of Food Safety - Mycotoxins in Foods
World of Food Safety 2013
Reinforcing quality and mitigating food safety risk to drive consumer confidence and profits
Overview of Presentation
Overview of mycotoxins in foods
Mycotoxin challenges in 21st century
Impact of mycotoxin on food industry and food regulations
Emerging analytical solutions for detection of mycotoxins
Strategies for ASEAN national food safety laboratories
Mycotoxins - Definition
The word mycotoxin stems from the Greek word "mykes" meaning mould and "toxicum" meaning poison.
Mycotoxins are secondary metabolites naturally produced by moulds (fungi) that may contaminant agriculture produce.
They can contaminate various agricultural commodities either before harvest or under post-harvest conditions.
Discovery of mycotoxins
Serious worldwide concern began in the early 1960s after
“Turkey X disease” was discovered in UK.
More than 100,000 young turkeys on poultry farms died in the
course of a few months.
Investigation of the early outbreaks showed that they were all
associated with feeds, namely Brazilian peanut meal .
Mycotoxins
Chemically and toxicologically diverse compounds
Aflatoxin B1
Carcinogenic to human (Group 1)
Patulin
Genotoxic
Fumonisin B1
Hepatotoxic, Nephrotoxic
Ochratoxin A
Potentially Carcinogenic (Group 2B)
Nephrotoxic
Citrinin
Zearalenone
Skin/membrane Irritant
Mycotoxins in Food
[1] Berthiller F., Sulyok M., Krska R., Schuhmacher R., Int. J. Food Microbiol. 2007; 119:33–37. [2] Environ Health Perspect. 2010 Jun;118(6):818-24. doi: 10.1289/ehp.0901388. Epub 2010 Feb 19
Today, 300-400 mycotoxins are known[1]. Aflatoxin B1 has been classified by the IARC to be a Class 1 human
carcinogen. A 2010 study by Liu, Y. Wu in collaboration with WHO estimated that
Aflatoxin causes between 5-30% of all liver cancer cases in the world[2].
Mycotoxins in Food There are six agriculturally important mycotoxins : aflatoxins,
trichothecenes, fumonisins, zearalenone and ochratoxin.
Mycotoxins Crops
Aflatoxins Groundnut, Maize, Almond, Fig, Pistachios, Hazenut, Sunflower seed, Sorghum, Cashew, Chestnut, Nutmeg, Rice, Chilli, Pepper, Turmeric, Milk
Ochratoxins Coffee, Grape, Paprika, Fig, Pepper, Barley, Nutmeg, Corn
Fumonisins Maize, Wheat, Rice
Trichothecenes Wheat, Oats, Corn
Zearalenone Corn, Wheat, Soyabean, Rice, Barley
Ergot alkaloids Rye, Barley, Wheat, Oats
References : RSAFF, Int. J. Mol. Sci., 2008, 9, pp2062-2090
Occurrence data (EFSA chemical occurrence database)
2,183 samples retrieved from EFSA chemical occurrence database on 15 Mar 13.
Samples collected between 2007-2012.
Analytical data on Aflatoxins (B, B1, G, G1).
Samples include cereal and milling products, processed cereal products.
Sampling carried out in 16 European countries.
Occurrence data (EFSA chemical occurrence database)
Distribution of total Aflatoxins by sampling year in cereals and milling products
Distribution of total Aflatoxins by sampling year in cereals and milling products
Occurrence data (EFSA chemical occurrence database)
Distribution of total Aflatoxins by sampling year in processed cereal products
Occurrence data (EFSA chemical occurrence database)
Mycotoxin occurrence
Field Crops Harvesting Drying/Silo storage
Feed mills Local farms Processing
Field mycotoxins contamination
Field mycotoxins contamination
Storage mycotoxins contamination
Storage mycotoxins contamination
Storage mycotoxins contamination
Storage mycotoxins contamination
Biological factors, Agronomic practices, Environmental factors(Temperature, Humidity, Time)
Mycotoxin contamination of meat/milk supply
Impact of Mycotoxins Significant economic losses are associated with their impact on
human health, animal productivity, and both domestic and international trade.
It is estimated that 25% of the world's food crops, including many basic foods, are affected by mycotoxin producing fungi.
According to FAO estimates global losses of foodstuffs due to mycotoxins are in the range of 1000 million tonnes per year [1].
Over $100 billion of exported commodities all over the world are susceptible to mycotoxin contamination [2].
[1] FAO wesbite (http://www.fao.org/food/food-safety-quality/a-z-index/mycotoxins/en/) [2] Cardwell K. F. (2001), Food and nutrition Bulletin, 21:488-492
Mycotoxin control - regulations Legislation is established in many countries worldwide. In Asia/Oceania, 26 countries have specific mycotoxin regulations.
Reference : Worldwide regulations for mycotoxins in food and feed in 2003
Mycotoxin regulations In Europe, ~ 99% of the 39 countries have specific mycotoxin regulations.
Reference : Worldwide regulations for mycotoxins in food and feed in 2003
Mycotoxin regulations The United States and Canada have five regulations on mycotoxins in 2003.
Reference : Worldwide regulations for mycotoxins in food and feed in 2003
Mycotoxin regulations Regulated levels of mycotoxins differ from nation to nation.
Reference : Worldwide regulations for mycotoxins in food and feed in 2003
Codex Standard 193-1995 Maximum limits exists for Total Aflatoxins, Aflatoxin M1,
Ochratoxin A and Patulin. Codex standards (Maximum limits) on 14 types of
foodstuffs EU Legislations on mycotoxins in foodstuffs Regulatory standards (Maximum limits) on 62 types of
foodstuffs
Mycotoxin regulations
Reference : JRC Technical Notes, Mycotoxin Factsheet, 2011
Codex Standard 193-1995
Commission Regulations (EC) No. 1881/2006
Commission Regulations (EC) No. 1881/2006
Commission Recommendation of 27 Mar 2013 (2013/65/EU)
Presence of T-2 and HT-2 toxins in cereal and cereal products
0 1 2 3 4 5 6 7
Aflatoxins B & G
Aflatoxin M1
Ochratoxin A
Patulin
Fumonisins
Deoxynivalenol
Zearalenone
Legislation established on regulatory limits of mycotoxins in ASEAN countries.
Mycotoxin regulations
Mycotoxin regulations Regulated levels of mycotoxins differ from nation to nation.
Country Limits for Patulin
Indonesia Fresh apple, canned apple, apple extract, nectar, alcoholic drinks
50g/kg
Apple puree
25 g/kg
Apple puree for infants and children 10 g/kg
Malaysia Apple juice (includes apple juice as ingredients in other beverages)
50 g/kg
Singapore Juices 10 g/kg
Vietnam Fruit and fruit juices 50 g/kg
Concentrated fruit juices and all product derived from them5
50 g/kg
Emerging Issues
(1) Masked Mycotoxins In mycotoxin contaminated commodities, many structurally
related compounds generated by plant metabolism or by food processing can co-exists with the native mycotoxin.
These derivatives may have very different chemical behaviour,
easily escaping routine detection. These derivatives can be hydrolysed to its precursors in the
digestive tract & can exert toxic effects comparable to the free form.
Emerging Issues February 2012 Glycoside derivatives of T-2 and HT-2 toxins in contaminated wheat and oats.
Emerging Issues “Occurrence of deoxynivalenol and deoxynivalenol-3-glucoside in durum wheat”, C. Dall'Asta, A. Dall'Erta, P. Mantovani, A. Massi, G. Galaverna, World Mycotoxin Journal, Dec 2012, pp. 83-91.
Abstract The occurrence of deoxynivalenol and deoxynivalenol-3-glucoside in durum wheat samples (n=150; 25 lines × 2 reps × 3 environments) collected in 2010 from 3 areas located in north-central Italy was evaluated. In addition, the co-occurrence of other trichothecenes was considered. An optimised extraction method based on the use of salts followed by ultra-high performance liquid chromatography-mass spectrometry analysis was used for the quantification of the mycotoxins. All samples were found positive for deoxynivalenol at concentrations ranging between 47 and 3,715 μg/kg. A ubiquitous occurrence of deoxynivalenol-3-glucoside was found; 85% of the analysed samples contained this masked mycotoxin at concentrations varying between 46 and 842 μg/kg. In addition to glycosylated deoxynivalenol, acetylated forms of deoxynivalenol (3- and 15-acetyldeoxynivalenol) were also found in most of the durum wheat samples. The deoxynivalenol-3-glucoside/deoxynivalenol ratio, reaching up to 30% in many samples, was similar to that already found in other cereals such as soft wheat and barley. These data open the way for further investigations on the role of glycosylating activity as a possible Fusarium head blight-resistance mechanism in durum wheat, as already proved in the case of soft wheat.
According to an article in Chemistry Research in Toxicology, current government limits on mold toxins in grain crops fail to account for so-called “masked mycotoxins,” which transform from harmless when outside the body to potentially harmful when inside.
Chiara Dall’Asta and colleagues say that plants protect themselves by conjugating glucose, sulfur or other substances to the mycotoxin, producing conjugated mycotoxins that are not harmful to the grain.
However, Dall’Asta’s research has shown for the first time that bacteria present in the human large intestine deconjugates deoxynivalenol (DON) and zearalenone (ZEN), the two most widespread mycotoxin contaminants. Once deconjugated, DON and Zen revert to their toxic forms.
http://www.foodengineeringmag.com/articles/90405-food-safety-regulations-miss-masked-mycotoxins
Emerging Issues
Ochratoxin A moieties T-2 /HT-2 toxins moieties Open-lactone-ochratoxin A 4-deoxy T-2 toxin Ochratoxin-A-hydrochinon 3-acetyl T-2 toxin 4R/4S-Hydroxy-ochratoxin A T-2 toxin 3-glucoside Ochratoxin alpha T-2 triol Ochratoxin alpha glucoronide T-2 tetraol Ochratoxin A glucuronide Glycoside derivative of T-2 11-Hydroxy-ochratoxin A Glycoside derivative of HT-2 Fumonisins moieties Deoxynivalenol moieties Hydrolysed B1 3 acetyl DON Hydrolysed B2 15 acetyl DON
Emerging Issues
Emerging Issues
Emerging Issues
Emerging Issues (2) Effect of climate change on mycotoxin geographic
distribution pattern EFSA’s Emerging Risks Unit identified changing patterns in
mycotoxin contamination in cereals such as wheat, maize and rice, due to climate change as a potential emerging hazard. In particular, aflatoxins (AFs) which are frequent in tropical and sub-tropical areas may become a concern in the EU.
With climate change and expected increasing temperature and
decreasing rain, the fungi, Aspergillus flavus and A. parasiticus may find conditions that are more suitable for their development.
Reference : “Modelling, predicting and mapping the emergence of aflatoxins in cereals in the EU due to climate change”, 18 Jan 2012, http://www.efsa.europa.eu/en/supporting/doc/223e.pdf.
Emerging Issues
+ 2oC climate change + 5oC climate change
Predictive modelling : Map of Aflatoxin B1 contamination risk (Maize) in the EU in different climate change scenarios
Emerging Issues (3) Emerging mycotoxins
Emerging Issues (3) Co-occurrence of multiple mycotoxins
Published 8 March 2013 139 mycotoxins & metabolites were detected in 83 feed samples.
All samples were co-contaminated with 7–69 different mycotoxins.
Emerging Issues (3) Co-occurrence of multiple mycotoxins
Beauvericin was found most often, in 98% of sample. Enniatins were second most, in 96% sample followed by DON and emodin, each 89%.
Major approaches for masked mycotoxin detection includes :
(a) Direct method using LC/MS/MS - Pros : Specific, Quantitative to target mycotoxin - Cons : Expensive, Lack of reference standards (b) Indirect method requiring acidic/enzymatic cleavage, LC/MS/MS strategies, etc. - Pros : Cheaper (?), Detection of uncharacterised conjugates - Cons : Not specific, not quantitative, Efficiency of hydrolysis
Analytical Strategies
Analytical Strategies Direct Approach
Analytical Strategies
Full scan mass spectra of deoxynivalenol-3-glucoside in ACN (1000 g/kg)
Analytical Strategies Direct Approach
Analytical Strategies
Analytical Strategies Indirect Approach
Analytical Strategies
Probing extracts for masked mycotoxin through investigation of parent ions chromatogram of product ions (m/z 185, 215)
Parent : 442
Parent : 604
Parent : 484
Parent : 646
Parent : 400
Flow injection ESI Product Ion Scan
T-2 glucoside HT-2 glucoside
Investigation of hidden mycotoxins
Analytical Strategies Indirect Approach
World Mycotoxin Journal, 2013
Flow Injection-Mass Spectrometry
Strategies for ASEAN National Food Testing Laboratories
Strategies for ASEAN National Food Testing Laboratories
Challenge 1 :
Strengthen the national food testing laboratory capacity to ensure safety of food supply particularly those of significance to ASEAN.
ASEAN Reference Laboratory
Field of Expertise Designated ARL
Mycotoxin Health Sciences Authority (HSA), Singapore
Pesticide Residues Agri-food and Veterinary Authority (AVA), Singapore
Heavy Metals & Trace Elements Department of Medical Sciences, Ministry of Public Health, Thailand
Veterinary Drug Residues in foodstuffs of animal origin
Department of Livestock Development, Ministry of Agriculture and Cooperatives, Thailand
Microbiology Quatest 3, MOST, Vietnam
Genetically Modified Organisms Department of Chemistry, Malaysia
ASEAN Reference Laboratory (1) Mycotoxin Training workshops Three rounds of Mycotoxin workshop organised for ASEAN national
laboratory staff in 2005, 2007 and 2009.
ASEAN Reference Laboratory (2) Mycotoxin Proficiency Testing Programme Annual PT was organised for all ASEAN national laboratories since
2010.
2010 Ochratoxin A in Cereals 2011 Aflatoxins B & G in Peanuts 2012 Aflatoxins B & G in Nutmeg Powder 2013 Aflatoxins B & G and Ochratoxin A in Chilli Powder
Strategies for ASEAN National Food Testing Laboratories
Challenge 2 : Development of testing methodologies suitable for determination of
mycotoxins in Asian food products.
Nutmeg Powder B1 – 1.8 g/kg B2 – 42 g/kg
Peanut B1 – 8.5 g/kg B2 – 1.8 g/kg
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Strategies for ASEAN National Food Testing Laboratories
Challenge 3 : Development of testing methodologies suitable for determination of
multiple mycotoxins in Asian food products.
Thank You for your attention.
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