1. National Center for Environmental Health Division of Laboratory Sciences Xiaoyun Ye, Xiaoliu Zhou, Josh Kramer, Lily Jia, Lee-Yang Wong, and Antonia M. Calafat National Center for Environmental Health Centers for Disease Control and Prevention Atlanta, GA 30341, USA Exposure to the Antimicrobial Agent Triclocarban in Convenience Samples of U.S. Adults

2. Triclocarban (TCC)  Antimicrobial agent used in consumer and personal care products  soaps  body wash  deodorants  cleansing lotion  toothpaste  plastics

3.  TCC is classified as a high production volume chemical requiring environmental risk assessment by the EPA. TCC is also regulated as an antibacterial ingredient under the FDA’s Tentative Final Monograph (59 FR 31402, June 17, 1994)  TCC has been detected widely in environment In surface waters, municipal waste water treatment effluents, sewage sludge, streams nationwide  Humans exposed to TCC because of use of this chemical in personal care products

4. Estimated Distribution of Active Ingredients in Consumer Antiseptic Hand and Body Washes by Product Form TCC and/or triclosan used in >2000 antimicrobial products in the US in 2014 Typical levels in soap range from 0.5% to 1.5 % (TCC) and 0.1% to 0.5 % (triclosan) http://www.fda.gov/downloads/aboutfda/reportsmanualsforms/reports/economicanalyses/u cm379555.pdf Antiseptic Active Ingredient Liquid, Gel, and FoamBar soap % of UPCs % of Total Equivalent (16 oz.) Units Sold % of UPCs % of Total Equivalent (16 oz.) Units Sold Triclosan93.199.87.90.98 Triclocarban0085.199.01 Chloroxylenol1.70.014.00.01 Benzalkonium Chloride0.40.000 Phenoxyethanol0.4000 Natural ingredients3.90.23.00.002 Total100

5. Adverse Effects Upon Exposure to TCC  Animal data suggest that TCC could impair mammalian reproduction  Act as endocrine disruptor by enhancing the action of testosterone and other steroids  Induce cross-resistance to antibiotic drugs used in human medicine FDA issued a proposed rule in 2013 requiring manufacturers to provide more substantial data to demonstrate the safety and effectiveness of antibacterial soaps

6. Exposure Assessment of Triclocarban by Biomonitoring  Assessment of internal dose by measuring the parent chemical (or its metabolite or reaction product) in human specimens  Considerations for conducting a biomonitoring study  Selection of biomarker  Matrix considerations  Sensitive and selective techniques  Data interpretation

7. Potential Biomarkers of TCC TCC oxidative metabolites (e.g., 2’-OH-TCC, 3’-OH- TCC) and their conjugates, N (N’)-glucuronide TCCs were identified in rats and human urine Ye XY, Zhou XL, Furr J, Ahn KC, Hammock BD, Gray EL, et al. 2011. Biomarkers of exposure to triclocarban in urine and serum. Toxicology 286: 69-74.

8.  Dermal contact is the major route of human exposure  Conjugated TCCs (N-glucuronide TCCs) were the main urinary metabolites  Although present, oxidative metabolites and their conjugates were minor products in humans Human exposure to TCC following 15 mins showering with a TCC-containing soap (0.6%) Schebb NH, etc., 2011. Investigation of Human Exposure to Triclocarban after Showering and Preliminary Evaluation of Its Biological Effects. Environ Sci Technol 45: 3109-3115.

9.  Archived specimens collected in 2011-2014 from a diverse group of US male (N=92) and female (N=104) volunteers with no documented occupational exposure to TCC  Participants provided urine anonymously. Except for sex information, no other personal or demographic data were available Urine samples for the study

10. Analytical Method  On-line solid phase extraction-high performance liquid chromatography-tandem mass spectrometry  Enzymatic deconjugation  Solid phase extraction  Chromatographic separation  Quantification using isotope dilution mass spectrometry

11. Deconjugation of N-glucuronide TCC  Hydrolysis of N-glucuronides could be different from those of O-glucuronides  Acid and enzymatic hydrolysis for deconjugation of N- glucuronides have been used  Relatively long incubation time and high concentration of enzyme might be required

12. Deconjugation of TCC by beta-glucuronidase from Helix pomatia, H-1  Original conditions in our regular method (left) Amount: 4 units/µL urine Incubation time needed: >30 hrs  Optimized conditions (right) Amount: 24 units/µL urine Incubation time needed: 4 hrs

13. Distribution of TCC concentrations (LOD=0.1 ng/mL) Sample SizeFrequencyMeanSelect %tile (ng/mL)Maximum of Detection (%)(ng/mL)50th75th95th(ng/mL) Total196558.10.10.663.1120 Male927215.40.316.685.3120 Female104411.6<LOD0.22.184.3

14. How do exposures to TCC and triclosan compare?  Males had significantly higher TCC concentrations than females (p <0.001 for T-Test on both concentration and Log [concentration])  Concentrations of TCC generally lower than of triclosan  Sex differences in concentrations of triclosan are not significant  Different applications of TCC and triclosan in PCPs, and different lifestyles for males and females may play a role Conc. (ppb)

15. How do exposures to TCC and other PCPs compare?  Benzophenone-3 is a sunscreen agent used extensively in PCPs  Compared to TCC, females’ BP-3 concentrations are higher than among males  Different lifestyle for males and females may explain differences Conc. (ppb)

16. Conclusions  US adults are exposed to TCC through use of antimicrobial personal care products  N-glucuronide TCCs can be used as biomarkers of exposure to TCC  Urinary concentrations of TCC are higher in males than in females, suggesting possible differences in lifestyle and usage of antimicrobial consumer products

17. Future Work  Include triclocarban in future NHANES to assess the exposure to this chemical in the general US population  Conduct studies to evaluate potential health effects associated with exposure

18. THANK YOU! For more information please contact Centers for Disease Control and Prevention 1600 Clifton Road NE, Atlanta, GA 30333 Telephone: 1-800-CDC-INFO (232-4636)/TTY: 1-888-232-6348 Visit: www.cdc.gov | Contact CDC at: 1-800-CDC-INFO or www.cdc.gov/info The findings and conclusions in this report are those of the authors and do not necessarily represent the official position of the Centers for Disease Control and Prevention. National Center for Environmental Health Division Name in this space