CONSUMPTION OF CONTAMINATED FOOD IS A MAJOR ROUTE OF HUMAN EXPOSURE TO TOXIC CHEMICALS, IN THE UNITED STATES AND GLOBALLY. ONE WAY THAT CHEMICALS UNINTENTIONALLY ENTER INTO FOODS IS THROUGH MIGRATION FROM FOOD PACKAGING. A CLASS OF CHEMICALS THAT HAS RECEIVED GROWING ATTENTION DUE TO HEALTH CONCERNS AND THAT IS HIGHLY RELEVANT FOR PACKAGED FOODS IS THE CLASS OF PER- AND POLYFLUORINATED ALKYL SUBSTANCES (PFAS). THESE CHEMICALS ARE WIDELY USED BECAUSE OF THEIR NON-STICK PROPERTIES AND THEIR ABILITY TO REPEL OIL. THEY ARE PRESENT IN BOTH PAPER-BASED AND PLASTIC-BASED FOOD PACKAGING, AND PEOPLE WHO CONSUME A HIGH PROPORTION OF PACKAGED FOODS, INCLUDING FAST FOOD AND TAKE-OUT FOODS, HAVE HIGHER CONCENTRATIONS OF THESE CHEMICALS IN THEIR BLOOD. SOME PFAS, SUCH AS PERFLUOROOCTANE SULFONATE (PFOS) AND PERFLUOROOCTANOIC ACID (PFOA) ARE NOW KNOWN TO BE BIOACCUMULATIVE AND TOXIC. THESE HAVE BEEN VOLUNTARILY WITHDRAWN FROM MUCH OF THE US MARKET, AND ARE RESTRICTED IN THE EUROPEAN UNION, BUT CONTINUE TO BE USED AND PRODUCED IN OTHER COUNTRIES. UNFORTUNATELY, THERE IS STILL A LACK OF INFORMATION ABOUT WHICH OTHER PFAS ARE PRESENT IN MOST PACKAGED FOODS, THOUGH A NUMBER OF PFAS-CONTAINING FOOD PACKAGING PRODUCTS ARE APPROVED BY THE FDA. THE CLASS OF PFAS INCLUDES MORE THAN 5,000 COMPOUNDS ACROSS A WIDE ARRAY OF INDUSTRIES, AND VERY LITTLE IS KNOWN ABOUT THE TOXICITY OF MOST OF THEM.IN THIS PROJECT, WE WILL PERFORM THE FIRST SYSTEMATIC SURVEY OF PFAS PRESENT IN BOTH DOMESTIC AND IMPORTED PACKAGED FOODS. WE WILL BUILD A DATABASE OF THE TYPES AND ORIGINS OF PACKAGED FOODS THAT ARE AVAILABLE IN BOTH NATIONAL SUPERMARKET CHAINS AND LOCAL INTERNATIONAL FOOD STORES, AND COME UP WITH A REPRESENTATIVE SELECTION OF SAMPLES FOR PFAS ANALYSIS. THE FOOD PACKAGING SAMPLES WILL BE TESTED FOR THEIR POTENTIAL TO IMPART PFAS TO FOODS THROUGH EXTRACTION (A MORE EXTREME CASE) AND THROUGH MIGRATION ASSAYS THAT SIMULATE ACTUAL CONDITIONS WHEN IN CONTACT WITH FOODS (A MORE TYPICAL CASE). USING STATE-OF-THE-ART METHODS, WE WILL ANALYZE THE MIXTURES THAT RESULT FROM EXTRACTION AND MIGRATION STUDIES TO IDENTIFY WHICH PFAS ARE PRESENT AND AT WHAT QUANTITY. BY EMPLOYING RECENTLY DEVELOPED NON-TARGET ANALYSIS TECHNOLOGY, WE WILL BE ABLE TO RESOLVE THE CHEMICAL STRUCTURES OF PFAS PRESENT EVEN IF THEY ARE PREVIOUSLY UNKNOWN AND/OR HAVE NEVER BEEN ANALYZED BEFORE. THIS NON-TARGETED ANALYSIS WILL BE FOLLOWED UP BY MORE TRADITIONAL HIGH-PERFORMANCE TARGETED ANALYSIS TO MEASURE THE CONCENTRATION OF PFAS THAT HAVE ANALYTICAL STANDARDS AVAILABLE. FINALLY, WE WILL ESTABLISH WHETHER THE TYPES OF PFAS PRESENT IN THESE FOOD PACKAGING SAMPLES AT THE CONCENTRATIONS DETECT POSE A RISK TO CONSUMERS BY EVALUATING THEIR POTENTIAL TO ACCUMULATE IN THE HUMAN BODY AND TO CAUSE TOXIC EFFECTS. THIS WILL BE DONE USING A COMBINATION OF COMPUTER SIMULATIONS AND EXPERIMENTS WITH ZEBRAFISH. THE SIMULATIONS WILL BE USED TO UNDERSTAND WHETHER THE PFAS WILL BIND TO KEY PROTEINS AND RECEPTORS IN THE BODY, LEADING FOR EXAMPLE TO ACCUMULATION IN THE LIVER OR TO CHANGES IN BLOOD CHEMISTRY. THE EXPERIMENTS ON ZEBRAFISH, AN ANIMAL MODEL OFTEN USED TO ANTICIPATE EFFECTS IN HUMANS, WILL DETERMINE AT WHAT DOSE THE PFAS CAUSE TOXICITY, AND IN PARTICULAR IF THERE ARE MEASURABLE DEVELOPMENTAL EFFECTS THAT COULD BE OF CONCERN TO VULNERABLE POPULATIONS SUCH AS CHILDREN.BASED ON THE OUTCOMES OF THIS PROJECT, BETTER POLICY AROUND THE USE OF PFAS IN FOOD PACKAGING CAN BE DEVELOPED, REGULATORS WILL BE MADE AWARE OF THE TYPES OF FOOD IMPORTS MORE LIKELY TO CONTAIN TOXIC PFAS, CONSUMERS WILL HAVE GUIDANCE ON WHAT SPECIFIC TYPES OF PACKAGING TO AVOID IN ORDER TO REDUCE THEIR EXPOSURE TO TOXIC CHEMICALS, AND SCIENTISTS WILL HAVE BETTER INFORMATION ABOUT HOW SPECIFIC PFAS STRUCTURES AND OR MIXTURES OF PFAS LEAD TO INCREASED TOXICITY, INFORMING THEM IN HOW TO BETTER DESIGN LESS HAZARDOUS CHEMICALS.