1. Toxicology Component of FDA’s Action Plan for Acrylamide Richard Canady, PhD DABT US Food and Drug Administration Center for Food Safety and Applied Nutrition www.cfsan.fda.gov Food Advisory Sub-Committee Meeting December 4 and 5, 2002

2. Goal of the toxicity component of FDA’s action plan Examine the likelihood that adverse health effects are caused by exposure to acrylamide-containing foods.

3. Context for toxicology research Ongoing assessment of knowledge base for acrylamide Past assessments for Food contact materials, cosmetics, water treatment, pesticide inert ingredient, specialized grout, monomer/polymer manufacture, coal mining, cigarette smoke Knowledge has changed with regard to How we are exposed Ingestion from food, not just water The levels to which we are exposed

4. Data gap evaluations World Health Organization/ Food and Agriculture Organization (WHO/FAO) consultation in June Interagency meeting in September Joint Institute for Food Safety and Applied Nutrition (JIFSAN) workshop in October Expert panel at Emerging Neurotoxicology Conference in November Workshop for germ cell mutagen risk assessment Spring 2003

5. Outline of presentation of what we know, need, and are planning 1. Toxicokinetics 2. Animal carcinogen 3. Human neurotoxicant 4. Reproductive/developmental effects 5. Safety/risk assessment

6. 1. What we know for toxicokinetics Absorbed orally Degree of absorption from food has not been studied Most toxicity information from drinking water or injections Wide and uniform distribution (oral exposure, high dose) Metabolism understood for >1000 micrograms per kilogram of body weight (mcg/kg) doses Saturable (CYP2E1) conversion to glycidamide Glutathione conjugation Elimination occurs in hours to days But protein binding (-SH) and potential for cumulative damage Exposure duration affects neurotoxic dose

7. 1. Toxicokinetics data needs Bioavailability of acrylamide from food Dose-response for toxicity, disposition, and binding Dose metrics, critical events, mode of action DNA to hemoglobin adduct relationship DNA and protein binding as a marker of toxicity and risk Significant nuclear protiens Toxicokinetics of acrylamide in humans Risk factors for susceptibility

8. 1. Plans for Toxicokinetics data development FDA – National Center for Toxicological Research (NCTR) DNA/protein adduct characterization and their relationship to effects and relevant external dose levels Bioavailability CDC – National Center for Environmental Health (NCEH) Biomarker-intake relationship in studies prior to NHANES Acrylamide monomer industry Human dosing study, PbPk model development

9. 2. What we know for cancer 2-year rat studies show cancer Drinking water exposures, high doses Epidemiology not sufficient to change conclusions or weight of evidence

10. 2. Carcinogenicity data needs Cancer epidemiology in populations of known high exposure Chronic toxicity/carcinogenicity study Acrylamide and glycidamide in rat and mouse Review histology slides from existing bioassays using updated diagnostic criteria Investigate the mechanism of thyroid tumor induction as reported in existing bioassays

11. 2. FDA carcinogenicity study plans: intermediate to long term FDA nominated acrylamide and glycidamide to the National Toxicology Program as priority selections Chronic carcinogenicity and mechanism NCTR will conduct these studies FDA will participate in all experimental protocol designs to assure regulatory needs are met

12. 2. FDA carcinogenicity study plans shorter term Mechanistic studies, including bioavailability and markers of exposure and effect DNA and protein adducts caused by acrylamide Adducts are reaction products between a chemical and either DNA or proteins Adducts can tell us How much exposure occurs and Help us understand the toxicology Adducts may be particularly useful in relating animal toxicity studies to potential risks for humans from acrylamide

13. 3. Neurotoxicity High dose exposures in occupational settings causes neurotoxicity in humans Effect widely studied, multiple species Cumulative dose important Age-related effects One study showed more rapid onset of neurotoxicity in young vs old animals Another showed greater response for neurotransmitter effects in younger animals But very little data overall

14. 3. Neurotoxicity data needs Further evaluation of interaction between dose and duration Improve weight-of-evidence for or against neurodevelopmental effects at food-acrylamide doses Establish a NOAEL for relevant endpoints

15. FDA – plans are being developed for Inclusion of neurotoxicity endpoints in NTP study Study of neurodevelopment Ongoing academic research into mechanism of neurotoxicity 3. Plans for neurotoxicity study

16. 4. Reproductive/ developmental effects (other than neurotoxicity) Decreased body weight, but no structural malformations Reduced litter sizes (5000 mcg/kg treatment of males) Germ cell mutagen in animals at high, injected doses Genetic effects in mice (injection >40,000 mcg/kg) Specific-locus mutations Heritable or reciprocal transformations Unscheduled DNA synthesis in spermatids,chromosomal aberrations or micronuclei frequency in spermatogenic cells

17. 4. Reproductive/ developmental effects data needs Epidemiology for germ cell toxicity Expression profiling for genotoxic effects on somatic and germ cells Define the germ cell mutations generated Dose-response Low dose and multiple dose exposure effects Compare food and drinking water exposures Mechanistic Adduct formation with DNA and significant nuclear proteins Dominant lethal study in CYP2E1 knockout mice

18. FDA Genotoxicity endpoints in NTP study, including initial subchronic studies (under development) Workshop for germ cell mutagen risk assessment NIOSH worker studies NIEHS collaboration for reproductive/genotox NIEHS evaluation of CYP2E1 knockout mice for dominant lethal effect Expert review panel by National Toxicology Program (NTP) Center for the Evaluation of Risks to Human Reproduction (CERHR) 4. Plans for Reproductive/ developmental effects study

19. 5. Safety/risk assessment USEPA RfD and USFDA ADI EPA Reference dose and FDA Acceptable Daily Intake based on Burek et al 1980 finding of sciatic nerve degeneration (electron microscopic) 90 day study in rats Lowest observed effect level 1000 mcg/kg No observed effect level 200 mcg/kg Only 3 animals examined at EM level per dose group Recovery seen at 144 days for LOAEL dose 1000 fold uncertainty factor Routine EPA re-evaluation of RfD underway

20. 5. WHO/FAO Safety/risk assessment Noncancer effects judged unlikely at doses from food Cancer No consensus on quantification of risk Carcinogenic potency characterized as …similar to that of other carcinogens in food (benzo[a]pyrene and heterocyclic aromatic amines) …” However, “… intake levels for acrylamide are likely to be higher.”

21. WHO/FAO conclusion “major concern” for acrylamide based on relative cancer potency and uncertainty regarding germ cell mutagenicity findings.

22. 5.Safety/risk assessment - Effective dose, Exposure, ADI Lowest neurotoxic dose in rats 1000 mcg per kg body weight No effect seen at 200 mcg/kg WHO/FAO – typical dietary intake 0.3-0.8 mcg/kg body weight FDA Acceptable Daily Intake for food contact decisions 1000 fold uncertainty factor 0.2 mcg/kg body weight

23. Summary Acrylamide causes effects in animals and in humans at doses much higher than those we get through food However, safety/risk assessment indicates the need for further analysis of the risk There are substantial gaps in our knowledge of whether the acrylamide levels in food are likely to cause health effects Data gap analyses by leading experts in a variety of venues in the last 7 months indicate what kind of information is needed We have initiated programs to carry out the needed information gathering and research, and are already seeing results