Monitoring the Air for Toxic and Genotoxic Compounds

Kim Rogers

Office of Research and DevelopmentNational Exposure Research LaboratoryU.S. Environmental Protection Agency

Why Are We Interested in Monitoring Toxic Vapors ?

• Industrial ContaminationDeliberateAccident

• Homeland Security

Why Toxicity Monitoring ?

• Simple• Rapid• Sensitive• Adaptable from Commercial Sector• Biochemical / Physiological Relevance

What Compounds Are of Interest ?

• Toxic Industrial Chemicals (TICs)High Risk

Toxic GassesMedium Risk

Toxic Volatiles

• Chemical Warfare Agents (CWAs)• Non-Traditional Agents (NTAs)

Adapt Waste Water Screening Assays for Air Monitoring

• Acute ToxicityMicrotoxIQ-Tox Daphnia magna

• GenotoxicityDNA Melting/Annealing Analysis

DMSO

ExposureChamber

SPMD

NeatTIC

Stockin

Methanol

Direct Microtox Assay

EC50

NeatTIC

Dilutionin Microtox

AssayReagent

Stockin

DMSO

AppEC50

Vapor Accumulation Assay

Dilutionin Microtox

AssayReagent

SPMDHow Does It Work

• Low Density Polyethylene– 10A pores

• Biological Mimic– Triolein, DMSO, Carbon

• After Deployment– Direct assay – Extract with solvent

SPMD in 40 mL and 4L Exposure Chambers

Microtox Reagent (bacterium)Photobacterium vibrio

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 170

50

100

150

200

250

300

Dik

eten

eP

hosp

horu

s ox

ychl

orid

eA

crol

ein

Tric

holo

race

tyl c

hlor

ide

Met

hane

sulfo

nyl c

hlor

ide

Stil

bene

1-O

ctan

ethi

olS

ulfu

ryl c

hlor

ide

Form

alde

hyde

Ally

lam

ine

Met

hyl c

hlor

ofor

mat

eC

hlor

oace

tone

Met

hylc

hlor

osila

neD

iisop

ropy

l flu

orop

hosp

hate

Met

hylh

ydra

zine

Ace

tone

cya

nohy

drin

1,2-

Dib

rom

oeth

ane

EC-5

0 (p

pmv)

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 160

100

200

300

400

500

600

700

800

Pho

spho

rus

oxyc

hlor

ide

-- 5

,400

Dik

eten

eFo

rmal

dehy

deM

ethy

lhyd

razi

neM

etha

nesu

lfony

l chl

orid

eA

llyla

min

eS

tilbe

neA

crol

ein

Tric

hlor

oace

tyl c

hlor

ide

Diis

opro

pyl f

luor

opho

spha

te

Sul

fury

l chl

orid

eM

ethy

lchl

oros

ilane

Met

hyl c

hlor

ofor

mat

eA

ceto

ne c

yano

hydr

in1,

2-D

ibro

moe

than

eC

hlor

oace

tone

1-O

ctan

ethi

ol

Con

cent

ratio

n Fa

ctor

s

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 170

200

400

600

800

1000

Phos

phor

us o

xych

lorid

eAc

role

inD

iket

ene

Tric

hlor

oace

tyl c

hlor

ide

1-O

ctan

ethi

olS

tilbe

neSu

lfury

l chl

orid

eM

etha

nesu

lfony

l chl

orid

eC

hlor

oace

tone

Met

hyl c

hlor

ofor

mat

e

Met

hylc

hlor

osila

neA

llyla

min

eD

iisop

ropy

l flu

orop

hosp

hate

Form

alde

hyde

Acet

one

cyan

ohyd

rin1,

2-D

ibro

moe

than

eM

ethy

lhyd

razi

ne

Appa

rent

EC-

50 (p

pbv)

123456789

10111213141516171819

-60 -40 -20 0 20 40 60 80

More Effective with 15 min. Assay

More Effective with 5 min. Assay

Zinc SulfateDiketeneAcrolein

Acetone cyanohydrinChloroacetone

MethylhydrazineFormaldehyde

AllylamineListerine

Stilbene1-Octanethiol1,2-DibromoethaneMethylchloroformateMethylchlorosilanePhenolHypochlorite (bleach)SulfurylchlorideMethanesulfonylchlorideTrichlororacetylchloride

% Relative Change in EC50 Between 5 and 15 min.

X A

xis

Title

0 5 10 15 20 25

0

20

40

60

80

100

% Toxicity % Mass

Rel

ativ

e M

ass

and

Toxi

city

(%)

Acro

lein

Accumulation Time (hr)

Daphnia magna Used in the IQ-Tox Test

20

40

60

80

100

sulfu

ryl ch

loride

methylc

hlorof

ormate

methylc

hloros

ilane

1,2-di

bromoe

thane

trichlo

roace

tyl ch

loride

allyla

mine

methan

esulf

onyl

chlor

ide

dikete

ne

acrol

ein

chlor

oace

tone

stilbe

ne

octan

e thio

l

phos

phoru

s oxy

chlor

ide

EC

-50

(ppm

v)

% Daphnia IQ-tox 1 hr % Daphnia 18 hr % Daphnia 24 hr

0.1

1

10

100

1,2-di

bromoe

thane

methylc

hlorof

ormate

methylc

hloros

ilane

sulfu

ryl ch

loride

trichlo

roace

tyl ch

loride

allyla

mine

methan

esulf

onyl

chlor

ide

dikete

ne

acrol

ein

chlor

oace

tone

stilbe

ne

octan

e thio

l

phos

phoru

s oxy

chlor

ide

EC

-50

(ppm

v)

% Daphnia IQ-tox 1 hr % Microtox 5 min

020406080

100120

0 0.02 0.04 0.06 0.08

Time (a.u)

Fluo

resc

ence

(fl.u

.)

020406080

100120

0 0.02 0.04 0.06 0.08

Time (a.u)

Tem

pera

ture

(o C)

10 50 100 150 200 2500

20

40

60

80

100

Concentration of DNA (ng/mL)

Relative Fluorescence (%

)

Control Cyclohexane Toluene Phenol Glutaraldehyde Mitomycin C0

20

40

60

80

100R

elat

ive

Fluo

resc

ence

(%)

Genotoxic and Non-Genotoxic Controls

ControlAcrolein

Allylamine

Chloroacetone

Formaldehyde

Acrylonitrile

Bromoethane

Crotonaldehyde0

20

40

60

80

100R

elat

ive

Fluo

resc

ence

(%)

Toxic Industrial Chem icals (100 m M )

Control

Acrolein

Allylam ine

Chloroacetone

Form aldehyde

Acrylonitrile

Brom oethane

Crotonaldehyde0

2 0

4 0

6 0

8 0

1 0 0Pe

rcen

t dec

reas

e in

fluo

resc

ence

T o x ic In d u s tr ia l C h e m ic a l (1 0 0 m M )

Control 10 20 40 60 80 1000

20

40

60

80

100

Relative Fluorescence (%

)

Concentration of Formaldehyde (mM)

0 20 40 60 80 1000

20

40

60

80

100R

elat

ive

Fluo

resc

ence

(%)

Concentration of Formaldehyde (mM)

Summary

• Air Sampling• Toxicity

MicrotoxIQ-Tox

• GenotoxicityDNA Melting Annealing Analysis

Future Directions• Sampling

– Carbon Fabric• Sensitivity

• Bioassay– Genetically Engineered Yeast

• Cytotoxicity & Genotoxicity

• Environmental Pollutants– Nanaomaterials

• Environmental Applications– Vapor, Water, Sediment

Acknowledgements

• Kumar Ramanathan NRC-Postdoc• Srividia Kailasam NRC-Postdoc• Stacey Harper EPA-Postdoc

• Jim Petty USGS, CERC, MO• Jim Huckins• Robert Gale