Transfer of pesticides from commodities to natural flavors and aroma chemicals Ian Ronningen

1. Transfer of pesticides from commodities to natural flavors and aroma chemicals Ian Ronningen Chem 4101 December 9, 2011

2. Flavors and Fragrances • Aroma chemicals are extracted from commodities through a number methods • Mechanical pressing • Solvent extraction • Distillation • Expression • These extracts are commonly concentrated and then sold as fragrances or flavors Diazinon Atrazine Warfarin

3. Hypothesis • Extraction processes commonly used to separate flavors and aroma chemicals from commodities also extract pesticides • Extracted pesticides could make their way to consumers either through foods (flavors) or fragrances (aroma chemicals) • Pesticides present have the potential to pose either a chronic or acute health risk Orange, Source of Expressed orange oil(1) Potential Final Application (2) Muzaurieta, Annie Bell. "Vitamin C Foods." The Daily Green. Web. 4 Dec. 2011 "OMG! Cookies." Poached Prose. Web. 04 Dec. 2011. <http://poachedprose.wordpress.com/2010/08/02/omg-cookies/>

4. Relevance • To gain a competitive advantage flavor houses are increasingly sourcing rare commodities which come from countries that generally have less regulation surrounding pesticides • Commodity testing is limited, and screens limited number of pesticides. Screening focus is towards pesticides used in the United States. • If any pesticide ended up harming a consumer the producer of the food or fragrance would see massive economic backlash Borneo Rainforest(1) Alachor 1. Borneo Rainforest - Borneo Malaysia." Online Travel Guides of Travel Destinations -. Destination 360.

5. Analytical Techniques

6. HPLC-MS/MS Analysis • High sample throughput, automated, rapid analysis, ability to deal with complex matrices • Mass Spectrometer has high analytical selectivity • Established Mass Spectra Libraries simplifies data analysis • Many stationary phases available for wide application • C18 is a good starting point, Phenyl Hexyl also an option • Online SPE Also an option for higher sample throughput

7. Experimentation Incoming Commodities Washing Final Product Extraction Sample and analysis Sample and analysis Sample and analysis Sample and analysis Addition of internal standards of phenolic butyl-parabens Control Experiments: • Sample organic or hydroponic commodities (for incoming and extraction, and final product) • Sample unused wash water "15 Eccentric Endangered Trees, Plants, and Flowers | WebEcoist." WebEcoist. Web. 07 Dec. 2011 "Citrus Processing." Triowin. Web. 07 Dec. 2011

8. HPLC-MS/MS Analysis Sample preparation and clean up Sample Separation and characterization RestekQuEChERSsSPE tubes are an AOAC Method for Prep. Available (1,2). Include: • C18, MGSO4, PSA, GCB • Reference to established libraries • Unidentified peaks will be further investigated Morphed, James, and Peter Hancock. “A RAPID METHOD FOR THE SCREENING AND CONFIRMATION OF OVER 400 PESTICIDE RESIDUES IN FOOD“ Waters Application Notes. Tech. Waters, 2008. Web. 1 Dec. 2011 AOAC Official Method 2007.01, Pesticide Residues in Foods by Acetonitrile Extraction and Partitioning withMagnesium Sulfate QuEChERS-A Mini-Multiresidue Method for the Analysis of Pesticide Residues in Low-Fat Products. ResTek Application Notes.

9. HPLC-MS/MS Concept HPLC Separation of compounds from matrix based off interaction of compounds with stationary and mobile phases Diagram of Separation (1) Mass Spectrometer Ionization of analytes happens at the source, MS can either be used to scan or it can be operated in MS/MS utilizing collision cell and second quadrapole to analyze fragmentation patterns Diagram of Triple-quad (2) “HPLC Separation Modes” Waters Application Notes, Waters Corporation. 2011. “Mass Spectrometry Introduction." Department of Chemistry , University of Pittsburgh.

10. Figures of Merit (1)Xu, Y. "Simultaneous Determination of Flonicamid and Its Metabolites in Vegetables Using QuEChERS and Reverse-Phase Liquid Chromatography Tandem Mass Spectrometry." Journal of chromatography. (2011). *Figure and requirements depends on the analyte, since instrument response and level in extract is dependent on pesticide

11. Summary • Sampling along the line of extraction of aroma chemical from commodities will allow investigation of safety of these products • HPLC-MS/MS Analysis allows for effective separation of pesticides from complex matrix and effective analysis of pesticides in natural flavors and fragrances extracted from commodities • Although added cost of a MS/MS instrument is high added selectivity and sensitivity justify cost

12. Additional References • AOAC Official Method 2007.01, Pesticide Residues in Foods by Acetonitrile Extraction and Partitioning with Magnesium Sulfate. • Culliney, T W. "Pesticides and natural toxicants in foods." Agriculture, ecosystems & environment 41.3-4 (1992):297-320 • Davies J, Barquet A, Vaclavek C, et al. Human Pesticide Poisonings by a Fat-Soluble Organophosphate Insecticide. Archives of Environmental Health. December 1975;30(12):608. • Picó, Y. "Environmental and food applications of LC–tandem mass spectrometry in pesticide‐residue analysis: An overview." Mass spectrometry reviews 23.1 (2003):45. • Reineccius, Gary. Flavor Chemistry and Technology. Boca Raton : Taylor & Francis, c2006. • Schreiber, A. "Application of spectral libraries for high-performance liquid chromatography-atmospheric pressure ionization mass spectrometry to the analysis of pesticide and explosive residues in environmental samples." Journal of chromatography 869.1-2 (2000):411 • Seiber, J N. "Contributions of Pesticide Residue Chemistry to Improving Food and Environmental Safety: Past and Present Accomplishments and Future Challenges." Journal of agricultural and food chemistry 59.14 (2011):7536-7543. •  Thurman, E M. "Choosing between atmospheric pressure chemical ionization and electrospray ionization interfaces for the HPLC/MS analysis of pesticides." Analytical chemistry 73.22 (2001):5441.