1. the World Mycotoxin Forum, 14-15 May 2001, the Netherlands 1 Parallel session 4 Recent analytical developments T. Nowicki Canadian Grain Commission, Canada

2. the World Mycotoxin Forum, 14-15 May 2001, the Netherlands 2 Acceptance of methods need methods that meet requirement for user friendliness and reliability many factors influence test results EC directives should be followed EC 98/53 outlines sampling and method guidelines for official control

3. the World Mycotoxin Forum, 14-15 May 2001, the Netherlands 3 Acceptance of methods (cont’d) precision parameters - look to ISO, AOAC, IUPAC, Codex use of Horwitz equation for evaluating repeatability and reproducibility performance parameters CEN report CP13505 sampling methods and performance criteria - 466/2001 EC

4. the World Mycotoxin Forum, 14-15 May 2001, the Netherlands 4 Acceptance of methods (cont’d) EC JRC has 8 institutes at 5 locations: - certified reference materials available from Geel Institute - see website (http://cpf.jrc.it) for training videos and method descriptions - information on EU law and legislation available at website (http://europa.eu.int/eur-lex and http://europa.eu.int/eur-lex/en/search_lif_simple.html)

5. the World Mycotoxin Forum, 14-15 May 2001, the Netherlands 5 Antibody based techniques traditional methods have drawbacks antibody based methods are reliable, sensitive, robust, rapid and cost effective good antibody required specificity requirements not always clear to manufacturers no antibody for patulin

6. the World Mycotoxin Forum, 14-15 May 2001, the Netherlands 6 Antibody based techniques (cont’d) IA columns; ELISA-plate, cards, tubes, biosensors many considerations in kit development recommendations to industry - correct sampling - provide reference matrices, technical assistance, training, standards, certified reference materials

7. the World Mycotoxin Forum, 14-15 May 2001, the Netherlands 7 Mycotoxin analysis in practice need sound and statistically valid sampling methods (EC directive 98/53) need to tailor extraction method and solvent to sample matrix and analyte cleanup - consider available commercial kits different detection technologies need different levels of cleanup

8. the World Mycotoxin Forum, 14-15 May 2001, the Netherlands 8 Mycotoxin analysis in practice (cont’d) consider possibilities for automation immunoaffinity columns - have disadvantages as well as advantages quality assurance is very important

9. the World Mycotoxin Forum, 14-15 May 2001, the Netherlands 9 Biosensors sensor surface with molecular recognition and signal transduction signal reduction with inhibitor multi-analyte detection CH3CN/water extraction with Romer Mycosep cleanup coatings include antibodies, enzymes, peptides

10. the World Mycotoxin Forum, 14-15 May 2001, the Netherlands 10 Biosensors (cont’d) detection limits e.g. aflatoxin B1, 0.2 ng/g strong correlation with GC and LC regenerable high sample throughput

11. the World Mycotoxin Forum, 14-15 May 2001, the Netherlands 11 Membrane based techniques alternative to expensive instrumental methods practical for portable field test developed by converting ELISA into test more suited for field use dipstick

12. the World Mycotoxin Forum, 14-15 May 2001, the Netherlands 12 Membrane based techniques (cont’d) flow-through format - semi-quantitative (cut-off levels) - colour/no-colour kits available (OTA, T-2, aflatoxin M1)

13. the World Mycotoxin Forum, 14-15 May 2001, the Netherlands 13 Near Infrared Spectroscopy no sample preparation time 1-5 min compared to GC-EC and HPLC correlation > 0.90 don’t know what molecule is being measured

14. the World Mycotoxin Forum, 14-15 May 2001, the Netherlands 14 Near Infrared Spectroscopy (cont’d) use of technology for measuring DON is feasible sensitive enough for proposed EU limit further improvement needed