Assessment of mycotoxins in grains and oilseeds most consumed in Ivory Coast D. Montet, L. Manizan, M. Oplatowska-Stachowiak, I. Metayer, D. Akaki, K. Campbell, C. Brabet didier.montet@cirad.fr
Major grains and oilseeds consumed in Ivory Coast Cereals and Oilseeds Commodities at high risk of contamination by mycotoxins Rice 84 % Rice Maize Peanuts Mil 2% Palm 7% Maize 3% Peanuts 4% According to survey of the 3C Ivoire UE project 2
Period and sampling places April 213 Main markets in Abidjan, Bouaké and Korhogo Samples (3-6 g) Total Rice Local rice 47 Imported rice 41 Total 88 Maize Cracked corn 29 Corn flour 5 Total 79 Peanut paste 71 Total 238 3
Type of samples Cracked maize Rice Peanut paste Maize flour with and without potash 4
Multi mycotoxin analysis by LC-MS/MS % Quantification of 79 mycotoxins RT control: 79*std+4IS (A=1 ng/ml) 2%DMSO in MeOH 14722_MO_13 2.8 1 XEVO-TQS#WAA788 22-Jul-214 13:4:48 2: MRM of 2 Channels ES+ TIC (Penicillic acid) 8.17e5 Time -..5 1. 1.5 2. 2.5 3. 3.5 4. 4.5 5. 5.5 6. 6.5 7. 7.5 8. 8.5 9. 9.5 1. 11 EU regulated mycotoxins: aflatoxins, OTA, FB1+FB2, ZEN, DON, Patulin 8 metabolites of EU regulated mycotoxins: 3-ADON, 15-ADON, de-epoxy-don, ZAN, α- ZEL, β-zel, α- ZAL, β-zal 6 masked mycotoxins: DON-3G, ZEN-14-G, α-zen-14g, β-zen-14g, ZEN-16-G, ZEN- 14-sulfate 54 emerging and other mycotoxins: T-2, HT-2, beauvericin, enniatins, equisetin, citrinin, fusaric acid, cyclopiazonic acid, sterigmatocystin Analyses carried out in collaboration with the Queen s University, Belfast, UK 5
EU regulation (1881/26, 165/21, 1126/27) fixing the maximum levels of mycotoxins in food intended for direct human consumption or use as an ingredient For rice, maize and their products Mycotoxins Maximal content AFB1 2 (cereals and derivatives) AFT 4 (cereals and derivatives) OTA 3 (cereals and derivatives) For peanuts and derivatives Mycotoxins Maximal content AFB1 2 AFT 4 FB1 + FB2 1 (maize and flour) ZEA 1 (maize and flour) 75 (other cereals and flour) 6
Toxicity of mycotoxins Toxins Toxicity TDI ((Tolerable Daily Intake) ng kg body weight/d Aflatoxins Hepatotoxic, genotoxic, immunotoxic, cancerogen AF B1: the most toxic of aflatoxins Group 1: human carcinogen (IARC) One of the most potent liver carcinogen No TDI Genotoxic carcinogenic effects without threshold DL5 (Lethal Dose Average) orally for mice mg/kg body weight 1 (AFB1) OTA Nephrotoxic, genotoxic, immunotoxic, neurotoxic, teratogen Group 2B: Possibly carcinogenic to humans (IARC) 18 (EFSA) 5 Fumonisins Lesion of the central nervous system, hepatotoxic, genotoxic, immunotoxic Group 2B: Possibly carcinogenic to humans (IARC) 2 (SCF) > 1 (FB1) Zearalenon Effects on fertility and reproduction, estrogen (menopause disorders) Group 3: Not classifiable as human carcinogen (IARC).2 (provisional, SCF) > 1 EFSA: European Food Safety Authority SCF: Scientific Committee of Food 7
% EU regulated mycotoxins % Local rice (47) Imported rice (41) 8 6 4 2 66 43 32 21 4 11 7 46 5 3 1-1 2 2 12 2 AFB1,12 2 4 1 14,1,15 2 4 1 2,9 Number of samples 11 8 1 2 18 1 OTA,12 3 1 15,1,31 3 7,9 Number of samples 8 1 1 4 1 8
% EU regulated mycotoxins 1 8 6 4 2 97 62 52 7 97 3 Cracked corn (29) AFB1,27 2 4 1 2 3 4 8,5 Number of samples 1 5 7 4 1 1 9
% EU regulated mycotoxins 1 8 6 4 2 96 56 5 3 2 88 1 Corn flour (5) AFB1,9 2 4 1 2 3 4 48,8 Number of samples OTA Number of samples 2 8 5 9 3 1 2 3,59 7,6 14 1 1
EU regulated mycotoxins %1 99 99 1 98 96 94 92 9 Peanut paste (71) AFB1 2 4 1 5 1 5 1,63 2 4535,5 Number of samples AFT 1 1 3 3 12 12 32 5 2 4 1 2 5 1 5 1 4 1,4 894,4 Number of samples 1 1 3 3 12 35 11 3 2 11
% Other mycotoxins 1 9 8 7 6 5 4 3 2 1 99 99 87 86 74 7 57 49 46 46 42 13 13 8 55 2 9 25 3 65 Rice Maize Peanut paste 12
Conclusions Peanut paste samples were the most contaminated, representing the highest risk to consumer health in CI With AFB1 and AFT levels up to 4535 µg/kg and 894 µg/kg, respectively Maize: up to 8.5 and 172.74 µg/kg for AFB1 and AFT, respectively Rice: up to 14.1 and 32.7 µg/kg for AFB1 and AFT, respectively % 96 1 99 99 Rice 1 91 9 8 Maize 7 57 58 6 52 5 Peanut 4 paste 3 24 18 22 17 18 2 3 6 8 1 1 13
Perspectives Study of the groundnut sector for peanut butter production Objectives Identification and understanding of abiotic and biotic factors responsible for mycotoxin contamination along the chain 14
Conclusion Under EU regulation Content in AFB1 in food 2 µg/kg Above mg/kg EU regulation Very top to EU regulation Sanitary risk Fatal risk Economic issues 15
Assessment of mycotoxins in grains and oilseeds most consumed in Ivory Coast Questions didier.montet@cirad.fr
Dynamic and biodiversity of toxigenic molds in connection with the production of aflatoxins and OTA Collecting samples along the peanut sector November 214, Korhogo region, Ivory Coast (grinding / homogenization) Sample preparation (crushing / homogenization) In France Aw Content in Aflatoxins and OTA by HPLC/FLD Enumeration and isolation of potentially producing aflatoxin fungi Identification and characterization of strains by conventional and molecular methods 17
Semis Récolte Gousses d arachide sur plants PC 1 PC 2 Séchage au champ une semaine Séchage au champ 2 à 3 jours Flow Diagram of Agriculteur : Production au champ et traitement post-récolte des PC 3 Séchage au village 3 à 4 jours Gousses d arachide séchées type 1 Séchage au village une semaine Gousses d arachide séchées type 2 the groundnut sector for the production of paste in the region of Korhogo gousses d arachide PC 4 Stockage and collection points (CP) Graines d arachide Décorticage Grossistes Stockage PC 5 Demi-grossistes et détaillants Stockage PC 6 Vendeuses de pâte d arachide Commercialisation des graines d arachide Grillage Retrait de la peau Arachide grillée 18
Type of samples and sampling locations Type of samples Abidjan Abobo Adjamé Yopougon Bouaké Korhogo Total Rice Local rice 9 9 9 9 11 47 Imported rice (dénikaschia) 9 9 9 9 5 41 Maize Cracked maize 6 7 7 5 4 29 Maize flour with potash Maize flour without potash 7 11 6 8 32 7 2 4 5 18 Peanuts Peanut paste 15 15 15 15 11 71 Total 53 4 53 48 44 238 19
Preparation of collected samples Crusher Retsch Homogenizer CHOPIN Blender Thermomix VORWERK 2
Other contaminant rice mycotoxins Local rice (47) Imported rice (41) Fumonisins OTB Beauvericin Citrinin Sterigmatocystin Equisetin Diacetoxyscripenol Fumonisins OTB Beauvericin Citrinin Sterigmatocystin Equisetin Cytochalasin B 21
Other contaminant peanut paste mycotoxins OTA et OTB Beauvericin Cyclopiazonic acid Diacetoxyscripenol Enniatin B Aurofusarin Equisetin Sterigmatocystin Acid 71 samples 22
Other contaminant maize mycotoxins Cracked maize (29) Flour with potash (32) Flour without potash (18) Beauvericin Citrinin Agroclavin Cyclopiazonic acid Equisetin Fusaric acid OTB Beauvericin Citrinin Agroclavin Cyclopiazonic acid Equisetin Fusaric acid OTB Beauvericin Citrinin Agroclavin Cyclopiazonic acid Equisetin Diacetoxyscripenol 23
Sample contamination rate Regulated mycotoxins % 9 8 7 6 5 4 3 2 1 83 56 49 32 2 37 5 238 samples rice, maize, peanut paste Other main mycotoxins Beauvericin, Equisetin, Citrinin, Fusaric acid, Cyclopiazonic acid, OTB, Sterigmatocystin, Cytochalasin B, Diacetoxyscripenol 24
% % Rice contamination rate Regulated mycotoxins 7 6 5 4 3 2 1 Local rice (47) 66 4 15 21 4 11 7 6 5 4 3 2 1 46 Imported rice (41) 12 2 2 2 AFB1,12 2 4 14 14,1 AFB1,15 2 4 14 2,9 Number of samples 11 8 11 1 Number of samples 18 1 OTA,12 3 1 15,1 OTA,31 3 7,9 Number of samples Number of samples 9 1 1 5 1 25
% % Rice contamination rate Regulated mycotoxins Local rice (47) Imported rice (41) 9 8 7 6 5 4 3 2 1 Abobo Adjamé Yopougon Bouaké Korhogo 9 8 7 6 5 4 3 2 1 Abobo Adjamé Yopougon Bouaké Korhogo AFB1 AFB1>2µg/kg AFT>4µg/kg OTA OTA>3µg/kg ZEA AFB1 AFB1>2µg/kg AFT>4µg/kg OTA OTA>3µg/kg ZEA Korhogo and Bouaké The most contaminated samples Bouaké The most contaminated samples 26
% Maize contamination rate Regulated mycotoxins 1 9 8 7 6 5 4 3 2 1 97 62 55 7 99 3 Cracked maize (29) AFB1,27 2 4 2 4 8,5 Number of samples 1 5 1 2 1 27
% % Maize contamination rate Regulated mycotoxins Flour with potash (32) Flour without potash (18) 1 9 8 7 6 5 4 3 2 1 94 47 34 13 3 81 13 1 9 8 7 6 5 4 3 2 1 1 72 72 55 1 6 AFB1,9 AFB1 2 4 2 4 48,8,32 2 4 2 4 57,1 Number of samples OTA 15 5 6 3 1 5 3 8 1 1 3,59 7,6 Number of samples Number of samples 3 1 28
% % % Maize contamination rate Regulated mycotoxins Cracked maize (29) 1 8 6 4 2 Abobo Adjamé Yopougon Bouaké Korhogo AFB1 AFB1>2µg/kg AFT>4µg/kg OTA OTA>3µg/kg FUM ZEA Flour with potash (32) Flour without potash (18) 1 1 8 8 6 6 4 4 2 2 Abobo Yopougon Bouaké Korhogo Abobo Yopougon Bouaké Korhogo AFB1 AFB1>2µg/kg AFT>4µg/kg OTA OTA>3µg/kg FUM ZEA AFB1 AFB1>2µg/kg AFT>4µg/kg OTA OTA>3µg/kg FUM ZEA 29
% Peanut paste contamination rate Regulated mycotoxins 1 1 99 99 71 samples 98 96 94 92 9 AFB1 AFB1>2µg/kg AFT>4µg/kg AFB1 2 4 5 1 5 1,63 4535,5 Number of samples AFT 1 1 18 12 34 4 1 4 5 1 5 1 4 1,4 894,4 Number of samples 1 8 11 36 11 3 1 3
% Peanut paste contamination rate Regulated mycotoxins 1 95 71 samples 9 85 8 75 Abobo Adjamé Yopougon Bouaké Korhogo AFB1 AFB1>2µg/kg AFT>4µg/kg Abobo and Adjamé 1% contaminated samples, at higher level than the European regulations 31
Other mycotoxins Average positives and maximum Mycotoxin Rice Maize Peanut paste Beauvericin.9 (11.8) 2.2 (22.5) 1.5 (11.) Equisetin 7.2 (114.8) 1.6 (4,7) 14.4 (139.9) OTB 1. (1.5).7 (1.1) 18.7 (259.9) Sterigmatocystin 1.9 (7.8) 8. (4.) Citrinin 42.3 (11.4) 212.8 (2394.1) Agroclavin 1.3 (3.5) Fumonisin B3 17.9 (82.6) Fusaric acid 25.3 (93.7) Cyclopiazonic acid 5.3 (18.8) 24.3 (131.7) Enniatin B.9 (2.6) Enniatin B1.7 (2.4) OTA 9.2 (147.4) 32