Biomonitoring Six Per- and PolyfluoroalkylSubstances (PFAS)

Lee Blum, Ph.D.

NMS Labs

Horsham, PA

HDC-SETAC

2019 Annual Spring Meeting

Princeton, NJ

April 24, 2019

2

I am an employee of NMS Labs, Inc., which is a bioanalytical toxicology laboratory that tests biological fluids and tissues for drugs and chemicals.

Disclaimer:

3

Outline

1. Introduction to PFAS

2. Method for biomonitoring

3. In-house dataa. NMS Labs’ population data of six PFAS

b. NMS Labs’ PFOA population data

4. Summary

4

Introduction

• PFAS are a class of synthetic chemicals widely used in industry and consumer products for their surfactant properties.

• They are environmental contaminants that can be found in areas where they are manufactured or where PFAS-containing products are used.

• These substances are found in fire-fighting foams, wetting agents, cleaners, stain repellents, and in surface finishes for textiles, paints, lacquers, and food packaging among other things.

5

Introduction

• Since PFAS are persistent chemicals, they easily move through the environment. As a result they are widely distributed and frequently found in sources of drinking water.

• In May 2012, the third Unregulated Contaminant Monitoring Rule (UCMR 3) from the U.S. EPA was published and required monitoring for 30 contaminants (28 chemicals and two viruses) between 2013 and 2015.

• including PFBS, PFHpA, PFHxS, PFNA, PFOA, and PFOS.

6

Hu XC et al., Environmental Science & Technology Letters, 2016.

7

2000: 263 3M workers for serum PFOA and PFOS (Olsen et al., 2003)

2004: 1025 DuPont workers for APFO (ammonium perfluorooctanoic acid) (Sakr et al., 2007)

2005-2006: 69,030 Ohio River Valley C8 Health Project (Frisbee et al., 2009).

2008-2009: 196 East Metro-Minnesota pilot study (Washington County, MN) (Minnesota Dept of Health, July 2009, Landsteiner et al., 2014)

2009: 153 residents around Decatur, AL (ATSDR, April 2013)

2015: 1578 residents in the vicinity of Pease Tradeport, Portsmouth, NH (NH DHHS, June 2016)

2016: 2903 residents in the vicinity of Hoosick Falls, NY for PFOA (NYDOH, June 2018)

1999-2016: National Health and Nutrition Examination Survey (NHANES) – CDC

National Report on Human Exposure to Environmental Chemicals

U.S. General Population Data

Biomonitoring U.S. Studies

8

Introduction

Health Effects of PFAS

From the ATSDR website:

Some, but not all, studies in humans with PFAS exposure have shown that certain PFAS may:

• affect growth, learning, and behavior of infants and older children

• lower a woman’s chance of getting pregnant

• interfere with the body’s natural hormones

• increase cholesterol levels

• affect the immune system

• increase the risk of some cancers

9Ref: http://digital.olivesoftware.com/Olive/ODN/PhiladelphiaInquirer/shared/ShowArticle.aspx?doc=PHQP%2F2018%2F12%2F23&entity=Ar00105&sk=3F0A8B0E&mode=text

10Ref: http://digital.olivesoftware.com/Olive/ODN/PhiladelphiaInquirer/shared/ShowArticle.aspx?doc=PHQP%2F2018%2F12%2F23&entity=Ar00105&sk=3F0A8B0E&mode=text

NMS Labs

Horsham, PA

11

12

Introduction

• Our objective was to develop a serum test for biological monitoring exposures to these same six PFAS.

13

Instrumentation – AB Sciex API 4500 QTrap tandem quadrupole mass spectrometer

14

Delay Column – Agilent Zorbax Bonus-RP Cartridge

Guard Column – Agilent Zorbax Bonus-RP Cartridge

Analytical Column – Agilent Poroshell 120 PFP

Mobile Phase A: Ammonium Bicarbonate

Mobile Phase B: Ammonium Bicarbonate in Methanol

Mobile Phase C: Formic Acid in DI Water

Mobile Phase D: Ammonium Formate in Methanol

Internal Standards

PFBS-13C3; PFHpA-13C4; PFHxS-13C3; PFNA-13C9; PFOA-13C4; PFOS-13C4

Method Parameters

15

Calibration

Analyte

AMR

(ng/mL) Slope Y-Intercept Correlation

PFBS 0.05 - 10.0 0.999 0.002 1.000

PFHpA 0.05 - 10.0 0.992 0.023 0.999

PFHxS 0.05 - 10.0 0.998 0.005 1.000

PFNA 0.05 - 10.0 1.007 -0.02 1.000

PFOA 0.5 - 100 0.993 0.195 1.000

PFOS 0.5 - 100 0.999 0.031 1.000

*Five replicate samples spanning the analytical range

16

Precision and Accuracy

Within Day 1

(n=5)

Between Day

(n=5 x 3 days)

Analyte

Target

Conc.

(ng/mL)

Accuracy

(%Diff)

Precision

(%CV)

Accuracy

(%Diff)

Precision

(%CV)

PFBS

0.05 3.5 5.1 4.2 3.9

0.25 -1.4 3.7 -3.6 3.1

6.00 -6.4 2.6 -4.9 2.1

PFHpA

0.05 8.1 5.1 -2.4 8.2

0.25 4.7 8.4 4.4 7.7

6.00 -7.8 2.4 -7.9 3.3

PFHxS

0.05 7.3 6.5 4.6 5.4

0.25 10.5 4.0 7.2 2.7

6.00 -8.2 2.9 -7.2 3.3

PFNA

0.05 16.8 7.0 6.7 6.0

0.25 -2.7 5.7 -3.3 4.8

6.00 -7.4 10.8 -6.7 6.2

PFOA

0.50 -9.6 8.5 -2.4 8.2

2.50 -8.1 4.3 -6.0 6.6

60.0 -5.8 3.3 -6.2 5.4

PFOS

0.50 6.1 3.3 4.3 2.9

2.50 -1.0 1.6 -0.3 3.6

60.0 0.3 1.0 -0.2 1.4

17

Chromatogram of 5/50 Calibrator

PFBS

PFNA

PFHpA

PFHxS

PFOS

PFOA

18

Chromatogram of 5/50 Calibrator

19

Chromatogram of Serum Sample

PFHpA

PFOA PFOS

20

Comparable Data for Six PFAS vs. General U.S. Population

* Not measured after 2013 - 2014

NMS Labs’ population data of random de-identified samples (n=151) with unknown exposures vs 2015 – 2016 U.S. CDC NHANES Data

PFBS

(ng/mL)

PFHpA

(ng/mL)

PFHxS

(ng/mL)

PFNA

(ng/mL)

PFOA

(linear isomer)

(ng/mL)

PFOS

(linear isomer)

(ng/mL)

NMS Population

Median <0.05 <0.05 0.99 0.42 1.12 1.77

NMS Population

97.5th% 0.06 0.47 5.75 1.44 4.13 11.5

2015 - 2016 US

CDC NHANES

Median

<0.10* <0.10* 1.20 0.60 1.50 3.20

2015 - 2016 US

CDC NHANES

95th%

<0.10* 0.20* 4.90 1.90 4.10 12.8

21

PFAS Panel Samples

* Not measured after 2013 - 2014

Patient samples of unknown exposures (n=36)

PFBS

(ng/mL)

PFHpA

(ng/mL)

PFHxS

(ng/mL)

PFNA

(ng/mL)

PFOA

(linear isomer)

(ng/mL)

PFOS

(linear isomer)

(ng/mL)

NMS Population

Median <0.05 <0.05 2.8 0.53 1.85 3.15

NMS Population

Range <0.05 <0.05 - 0.27 0.28 - 120 0.13 - 1.7 <0.05 - 120 0.78 - 17

2015 - 2016 US

CDC NHANES

Median

<0.10* <0.10* 1.20 0.60 1.50 3.20

2015 - 2016 US

CDC NHANES

95th%

<0.10* 0.20* 4.90 1.90 4.10 12.8

22

PFAS Panel Samples

* Not measured after 2013 - 2014

Patient samples of unknown exposures (n=36)

PFBS

(ng/mL)

PFHpA

(ng/mL)

PFHxS

(ng/mL)

PFNA

(ng/mL)

PFOA

(linear isomer)

(ng/mL)

PFOS

(linear isomer)

(ng/mL)

NMS Population

Median <0.05 <0.05 2.8 0.53 1.85 3.15

NMS Population

Range <0.05 <0.05 - 0.27 0.28 - 120 0.13 - 1.7 <0.05 - 120 0.78 - 17

2015 - 2016 US

CDC NHANES

Median

<0.10* <0.10* 1.20 0.60 1.50 3.20

2015 - 2016 US

CDC NHANES

95th%

<0.10* 0.20* 4.90 1.90 4.10 12.8

23

PFAS Panel Samples

* Not measured after 2013 - 2014

Patient samples of unknown exposures (n=36)

PFBS

(ng/mL)

PFHpA

(ng/mL)

PFHxS

(ng/mL)

PFNA

(ng/mL)

PFOA

(linear isomer)

(ng/mL)

PFOS

(linear isomer)

(ng/mL)

NMS Population

Median <0.05 <0.05 2.8 0.53 1.85 3.15

NMS Population

Range <0.05 <0.05 - 0.27 0.28 - 120 0.13 - 1.7 <0.05 - 120 0.78 - 17

2015 - 2016 US

CDC NHANES

Median

<0.10* <0.10* 1.20 0.60 1.50 3.20

2015 - 2016 US

CDC NHANES

95th%

<0.10* 0.20* 4.90 1.90 4.10 12.8

24

NMS Labs PFOA Population Data, 2014 - 2017

n

Ages: Avg ±SD, Median

(yrs)

AgesRange

(yrs)

PFOA Mean ± SD

(ng/mL)

PFOA Median (ng/mL)

PFOA95th%

(ng/mL)

PFOARange

(ng/mL)

PFOA Number

<0.5 ng/mL

PFOA Number

≥1000 ng/mL

Overall 2711 59 ± 15, 62 10 to 95 28.6 ± 268 5.8 62 <0.5 - 8200 29 10

Men 1282 60 ± 15, 63 10 to 91 42.3 ± 377 7.0 76 <0.5 - 8200 11 6

Women 1402 58 ± 15, 60 10 to 95 15.4 ± 87.5 5.0 47 <0.5 - 2200 18 3

NHANES (n-PFOA)2013 - 20142014 - 2016

21651993

12 to ≥2012 to ≥20

1.91.5

5.34.1

25

NMS Labs PFOA Population Data 2014 - 2017

(NMS)

(NMS)

26

27

Median Serum Concentrations of PFOA by Gender

* PFOA (linear)

7

1.83.45

31.9

5

1.32.85

25.4

0

5

10

15

20

25

30

35

NMS Labs* NHANES*(2015-2016)

Pease Trades,Portsmouth, NH

Hoosick Falls,NY

PFO

A S

eru

m (

ng

/mL)

Males Females

28

NMS Labs PFOA Testing: 2014 – 2018 (n = 3693)2015 - PFOA production and use stopped in U.S.

54.2

28.2

21.819.1

10.9

0.0

10.0

20.0

30.0

40.0

50.0

60.0

2014 2015 2016 2017 2018

NMS - PFOA Serum

PF

OA

Se

rum

(n

g/m

L)

2.081.94

1.56

0

0.5

1

1.5

2

2.5

2011 - 2012 2013 - 2014 2015 - 2016

PFO

A S

eru

m (

ng

/mL)

NHANES - PFOA Serum

29

Summary1. Six PFASs are tested in drinking water by the U.S. EPA under the third

Unregulated Contaminant Monitoring Rule. Method developed a method by LC/MS/MS to test for the same six PFASs in serum to use for biological monitoring of these substances.

2. The analysis of random serum samples with unknown exposures to the six PFASs demonstrated comparable findings to those observed in the general U.S. population.

3. In the samples tested by NMS Labs during 2014 – 2017 (n=2711), the overall PFOA serum concentrations appear to be higher than those values observed in the general U.S. population.

4. Serum PFOA concentrations appear to be higher in men than women.

5. Over the past 5 years (2014 - 2018) of testing (n = 3693), the average PFOA serum concentrations have declined in each subsequent year.

30

Thank you for your attention!