Testing for Bisphenol A and other Endocrine Disruptor Chemicals Lee Blum, Ph.D.

1. Lee Blum, Ph.D. Testing for Bisphenol A and other Endocrine Disruptor Chemicals

4. Bisphenol A NTP – The Study & The Results

5. Bisphenol A – It’s Everywhere

10. BPA - Instrumentation Hewlett Packard/Agilent 5973 MSD

12. BPA – Linearity in Urine Observed Concentrations (ng/mL) Target Value Day 1 Day 2 Day 3 10.0 ng/mL 10.6 10.4 10.2 5.0 ng/mL 4.90 5.02 4.99 2.5 ng/mL 2.44 2.43 2.50 1.0 ng/mL 0.98 0.98 0.98 0.25 ng/mL 0.25 0.25 0.25

13. BPA – Linearity in Urine

14. BPA - Within Day Precision in Urine N = 5 x 3 Days 0.25 ng/mL 0.5 ng/mL 7.5 ng/mL Day 1 Mean 0.30 ± 0.015 0.44 ± 0.01 6.64 ± 0.1 %CV 5.83% 1.26% 0.82% % Accuracy 104% 87.2% 88.5% Day 2 Mean 0.25 ± 0.026 0.41 ± 0.008 6.77 ± 0.1 %CV 10.4% 1.99% 1.69% % Accuracy 101% 82% 90.3% Day 3 Mean 0.23 ± 0.015 0.44 ± 0.02 6.38 ± 0.5 %CV 6.48% 3.59% 7.24% % Accuracy 93.6% 88.0% 85.1%

15. BPA - Between Day Precision in Urine N = 5 x 3 Days 0.25 ng/mL 0.5 ng/mL 7.5 ng/mL Mean 0.25 ± 0.02 0.43 ± 0.02 6.6 ± 0.30 %CV 8.91% 3.87% 4.63% % Accuracy 100% 86.0% 87.9%

16. Urine Sample Endogenous (ng/mL) Spiked (ng/mL) Target (ng/mL) Measured (ng/mL) Deviation from Target 1 0.15 1.0 1.15 1.12 -2.61% 2 1.75 2.5 4.25 4.05 -4.71% 3 2.61 5.0 7.61 6.87 -9.72% 4 0.41 10.0 10.4 9.26 -11.0% 5 3.89 15.0 18.9 19.0 0.53% 6 0.31 0.25 0.56 0.54 -3.57% 7 1.78 1.0 2.78 2.69 -3.24% 8 0.47 2.5 2.97 2.81 -5.39% 9 1.30 5.0 6.30 6.07 -3.65% 10 0.16 1.0 1.16 1.04 -10.3% 11 0.24 2.5 2.74 2.62 -4.38% 12 4.82 5.0 9.82 9.06 -7.74% 13 0.90 0.25 1.15 1.08 -6.09% 14 0.57 1.0 1.57 1.50 -4.46% 15 0.57 2.5 3.07 2.92 -4.89% 16 1.78 5.0 6.78 6.63 -2.21% 17 2.70 0.25 2.95 3.01 2.03% 18 4.28 1.0 5.28 5.26 -0.38% 19 1.08 2.5 3.58 3.06 -14.5% Mean Endogenous 1.57 ± 1.46 ng/mL Mean Bias -5.07 ± 4.15%

17. BPA – Storage Stability in Urine

18. BPA – Storage Stability in Urine

19. BPA – Storage Stability in Urine

20. BPA – Patient Urine Samples Samples Tested 36 Median 3.2 ng/mL Range <0.25 to >10 ng/mL

21. BPA – 0.25 ng/mL in Urine

22. BPA – Typical Urine Sample

24. BPA – Linearity in Serum

25. BPA – Linearity in Blood

26. BPA - Within Day Precision in Serum N = 5 0.25 ng/mL 0.75 ng/mL 7.5 ng/mL Mean 0.21 ± 0.01 0.80 ± 0.06 7.61 ± 0.26 %CV 5.12% 6.94% 3.42% % Accuracy 85.6% 106% 102%

27. BPA – Spiked Plasma Samples Plasma Sample Endogenous (ng/mL) Spiked (ng/mL) Target (ng/mL) Measured (ng/mL) Deviation from Target 1 0.22 0.25 0.47 0.47 0.00% 2 0.22 2.50 2.72 2.63 -3.31% 3 0.67 0.50 1.17 1.15 -1.71% 4 0.67 5.00 5.67 5.48 -3.35% 5 0.26 1.00 1.26 1.30 3.17% 6 0.26 10.0 10.3 10.0 -2.53% 7 0.11 0.75 0.86 0.92 6.98% 8 0.11 7.50 7.61 7.52 -1.18% 9 2.10 3.00 5.10 5.31 4.12% 10 2.10 9.00 11.1 11.3 1.62% Mean Endogenous 0.67 ± 0.83 ng/mL Mean Bias 0.38 ± 3.49 %

28. BPA – Spiked Blood Samples Blood Sample Endogenous (ng/mL) Spiked (ng/mL) Target (ng/mL) Measured (ng/mL) Deviation from Target NaF 0.08 0.25 0.33 0.36 9.09% NaF 0.08 2.50 2.58 2.86 10.8% K Oxalate 0.17 0.50 0.67 0.70 4.48% K Oxalate 0.17 5.00 5.17 6.06 17.2% Na EDTA 0.18 1.00 1.18 1.32 11.9% Na EDTA 0.18 10.0 10.2 12.2 19.4% K EDTA 0.12 0.75 0.87 0.83 -4.60% K EDTA 0.12 7.50 7.62 9.00 18.1% Na Heparin 0.22 3.00 3.22 3.30 2.48% Na Heparin 0.22 9.00 9.22 10.1 9.54% Mean Endogenous 0.15 ± 0.05 ng/mL Mean Bias 9.85 ± 7.54%

29. BPA – Storage Stability in Serum

30. BPA – 0.25 ng/mL in Serum

31. BPA – 0.75 ng/mL in Blood

34. [email_address] Phone: 1-800-522-6671 Lee Blum, Ph.D.