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T ab le of Co nte nts

A B S T

R

A C T

I I I

A C

K

N O

W

L

E D G

M

E N T S

I V

L I ST O F F I G U R E S

V

L I ST O F T A B L E S

V I

I N

T R O

D

U C T I O N

:

L I T E R A T U R E R E V I E

W

A N D B A C K G

R

O U N D

1

B

A

C

K

G

R

O U

N D

1

P A H

S

1

S

P

A

H

M

O

D

E

L

4

M

I

C

R

O

T

O

X -A

C U T E

6

M

IC R O T O X

-

C H R O N I C

1 1

O B J E C T I V E S

12

M

A T E R I A L S A

N

D

M

E

T

H O

D

S

14

C

H E

M

I C A L S

14

P A H S A

M

P L E

P R

E

P

A R A T I O N

14

M

I C R O T O X

-

A C U T E T O X I C I T Y

14

I N D I V I D U A L P A H T

E

ST I N G

15

P A H

M

I X T U R E T E ST I N G

16

C H R O N I C T O X I C I T Y

16

C H R O N I C T O X I C I T Y T E ST I N G

17

M

E D I A P R E P A R A T I O N

18

P O S IT I V E C O N T R O L P R E P A R A T I O N

18

S A

M

PL E P R E P A R A T I O N

18

T E S T P R O C E D U R E

19

Z P A H

M

O D E L

19

R E

S U L T S A N D D I S C U SS

I

O N

2 1

I N D I V I D U A L P A H A C U T E T O X I C I T Y T E S T S

2 1

(2)

D U A L

M

I X T U R E S

2 3

O T H E R

M

I X T U R E S

2

3

S P A H

M

O D E L

2 4

C H R O N I C T O X I C IT Y

2 5

C O N C

L U

S

I

O

N 2 7

R

E F E R E N C E S

2 9

(3)

A B ST R A C T

T

o

d

ate ,

f

ew r e s e a r c

h

er s

h

a v e a s s e s s e

d

t

h

e t ox

i

c

i

t

i

e s o

f P

o

l

ycyc

l i

c

A

r o

m

at

i

c

H

y

d

r ocar

b

ons

(

P A H

'

s

)

as

m i

xtu r e s, a

l

t

h

ou

g h

t

h

at

i

s

h

ow t

h

ey c om

m

o n

l y

oc c u r

i

n t

h

e env

i

r o n

m

e nt.

T h

e to x

i

c

i

t

i

es o

f

2,

3

, an

d 4

r

i

n

g

P A

H

'

s w e r e

d

ete r

m i

n e

d f

o r

i

n

d i

v

id

ua

l

c o

m p

ou n

d

s.

S

yn er

g i

st

i

c anta

g

o n

i

sts or a

d d i

t

i

ve e

f f

ect s o

f m i

x tu r es o

f

two , thr e e , a n

d f

o u r

P A H

c o

m p

ou n

d

s wer e a s s e s s e

d b y

c om

p

ar

i

n

g

t

h

e to x

i

c

i

t

i

e s of tho s e

m i

xtu r e s to t

h

at o

f i

n

d

iv i

d

u a

l

co m

p

o u n

d

s.

M

i

c ro t ox

m i

xtu r e re s ults we re a

l

s o c o

m p

are

d

t o t

h

o s e

p

r e

d i

ct e

d b y

t

h

e

I PA H m

o

d

e

l

.

T h

e t ox

i

c

i

t

i

e s o

f

c

h

r

y

s e n e ,

p y

r e n e ,

p h

e n a nt

h

ren e ,

f l

u o r ene ,

f l

u o r a nt

h

r e n e, ace n a

p h

t

h

a

l

e ne,

p h

e n a nt

h

r e n e, a n

d

n a

p h

t

h

a

l

e n e we r e ev a

l

u at e

d

us

i

n

g

t

h

e

M

i

croto x

®

a c ute a n

d

c

h

ron

i

c t ox

i

c

i

ty a ssay w

h i

c

h

u s e s a

b i

o

l

u

m i

n e s c e nt

b

ac te r

i

a. V

i b ri

o

f

i

s

h

e r

i

, to

m

eas u r e t

o x

i

c

i

t

y

. Re s u

l

t s

i

n

d i

c a te t

h

at, a s

i

n

d i

v

i d

u a

l

c o

m p

o u n

d

s, t

h

e a c ute t ox

i

c

i

t

i

e s o

f P A H

s a r e

i

n v e rse

l

y r e

l

ate

d

to wate r s o

l

u

b i l i

ty .

T h

e r e s e a r c

h

a

l

so s

h

o we

d

t

h

a t m

i

xtu r es c a n

b

e syn e r

g i

s t

i

c,

a ntago n

i

st

i

c, o r a

d d i

t

i

v e.

C h

r o n

i

c te sts s

h

ow e

d

t

h

a t

P A H

s w

i

t

h l

owe r wate r s o

l

u

b i

l

i

ty r e

q

u

i

re

g

r e ate r conc entra t

i

ons t

h

an o t

h

e r

P A H

s t o s

h

ow a to x

i

c r e s

p

o n s e.

L

a st

l

y

,

f

r o

m

the r e s e a r c

h

, o n e ca n s e e that t

h

e a

d d i

t

i

v e c o n c e

p

t

f

o r P

A

H

m

ixtu r e s

i

s

g

e n e r a

l l

y u

p

hel

d

.

(4)

A C K N O

W

L E D G

M

E N T S

I

wo u

l d f i

r s t

l i k

e to t

h

an

k G

o

d f

o r

g i

v

i

n

g

me t

h

e cour a

g

e an

d

d

ete r

m i

n at

i

o n to a c c o

m p l i

sh

m

y

g

o a

l

s. I

h

a v e to t

h

a n

k m

y

f

a

m i l

y a n

d f

r

i

e n

d

s

f

o r t

h

e

i

r

s u

p p

o rt a n

d

e n c o u ra

g

e

m

e nt a s wel

l

. E

l i

z a

b

et

h G

ut

h

e r

i

e a n

d C h

a

d

Ro

p

e r

h

el

p

e

d m

e s o

m

u c

h d

u r

i

n

g

t

h

e

l

a s t t w o

y

e ars.

T h

an

k y

o u so

m

u c

h f

o r e ver

y

t

h i

n

g

.

I

wo u

l d

a

l

s o

l i k

e

t o t

h

a n

k D

r .

P f

a en

d

e r

, w

h

o

b

e

l i

e v e

d i

n

m

e an

d

a

l l

o

w

e

d m

e to wo r

k w i

t

h h i m

.

(5)

L I ST O F F I G U R E S

F I

G U

R E 1. P

A H S

tr u ctu r e

,

W

ate r

So

l

u

b i l i

ty, a n

f L

o

f K

ow

F I G U R E 2

.

I

n

d i

v

i d

ua

l P A H T

ox

i

c

i

t

y E

x

p

re s s e

d i

n

T

ox

i

c

U

n

i

t s

F I

G U

R

E 3

. P

A H T

o x

i

c

i

ty v s

W

ate r

So

lu

b i l i

t

(6)

L I

S

T

O F T A

B L E

S

T A B L E

L

P

A

H T o xicty

U

n

i

ts o

f

In

d i

v

i d

u al

C

o

m

po u n

d

s

T A

B L E 2

.

D

u a

l

M

i

x tu r e s

T A B L

E 3

.

P A H T

ox

i

c

i

ty

U

n

i

t s o

f

M

i

xtu r es

T

A

B

L

E

4

. P A H

C h

r o n

i

c To x

i

c

i

t

y

(7)

As s e s s

m

ent of

C

hr onic an d A cute T o xici t y of Pol ycycl ic A r o

m

atic

H

y dr o c a rbo n s

(

P

A H

s

)

a n

d P A H

M

i

xtu r e s

I N T R O D U C T I O N A N D O B J E C T I V E S

I

ntr o

d

u ct

i

o n :

L i

teratu re rev

i

ew a n

d b

a c

k g

r ou n

d

PA

H

s

P A H

s are

m

u

l

t

i

-rin

g

str u ctu r e s

h

e

l d

t o

g

et

h

e r

b

y s ta

b l

e c a r

bo n

-car

bo n

b

o n

d

s

that can

b

e s u

b

st

i

tute

d

w

i

t

h

eit

h

e r n

i

tro

g

en , su

l f

u r , o r o x

y g

e n

(F i g

u r e

1

)

(

B l

u

m

e r,

19 7

6

)

.

T h

e

y

o c cu r

i

n t

h

e en v

i

ro n

m

e n t w

h

e n c o

m p l

ex or

g

an

i

c su

b

s t anc e s are ex

p

o s e

d

to

h i g h

t e

m p

er atu r es or

p

r ess u r e

(A g

e n cy

f

o r

T

o x

i

c

S

u

b

st an c e s a n

d

D

i

s e ase

R

e

g i

stry,

19 9 3

)

.

PA H

s are c o

m

mon c ont a

m i

na n t s o

f h

az a r

d

ou s wa st e s

i

t e s an

d

a re

p

r e s ent

in

ae r

i

al fa

l l

o ut o

f

co

m b

u stio n

p

r o

d

u ct s,

in

d

u str ia

l

a n

d

s ewa

g

e

le a

k

a

g

e, a n

d

t

h

e acc

id

e ntal

d is

p

os a

l

o

f p

etro

le u

m

p

r o

d

u cts

(

C

er n

ig l i

a,

19 92

)

. P

A H

s occur n atu ra

l l y i

n t

h

e

en v

ir

on

m

ent

d

ue t o o

i l

se e

p

s, su r

f

a ce r u n

-o

f f

, an

d f

o res t an

d p

ra

i

r

i

e

f i

re s,

b

ut t

h

e

i

r

c once ntr at

i

o ns c a n

b

e e

le v a

te

d d

u e t o

h

u

m

a n a c t

i

v

i

t

i

e s

(

C h

au n

d

r y,

199 5

)

.

P hy

s

io c

h

e

m ic a

l

c

h

a ract er

is

t

i

c s, su c

h

a s

lo

w w a te r s o

l

u

b il

i

t

y

an

d h ig h

o c t an o

l

-water coe

f f i

c

i

ents, a

f f

e c t t

h

e

p

ers

i

st en ce o

f

t

h

e s e c om

p

o u n

d

s

in

t

h

e en v

ir

on

m

ent.

P A H

s are n o t ver

y

s o

lu

b l

e , an

d

t

h

e

ir

wa te r s o

lu

b i l i

t

y

d

ecr e ase s w

i

t

h

in c r e

as

in

g

m

o

l

ec u

l

ar we

i

g

h

t.

P A H

s

h

ave a

h ig h

a

f f i

n

it

y f

or or

g

an

i

c

m

a t t e r an

d

or

g

an

i

c c o nt a

i

n

in

g

(8)

se

d

ime nt, w

h

er e t

h

ey pa rt

i

t

i

o n to t

h

e orga n

i

c

f

r a ct

i

on w h ic

h

c a n r edu c e their

b i

o a v a

i l

a

b i l i

t

y

.

T h

e

y

ar e

b

u r

i

e

d

a n

d p

e rs

i

s t u nt

i

l

d

e

g

r a

d

e

d

,

b i

oaccu

m

u

l

a te

d

, o r

re

m

ove

d b y d

re

d g i

n

g

(

C

er n

i g l i

a,

19 9 2

)

.

T h

er e

i

s t

h

ere

f

or e a

d i

r ec t re

l

a t

i

o ns

h i p

b

etwee n

P A H

to x

i

c

i

t

y

a n

d b i

o a v a

i l

a

b i l i

ty . P

A H d

e

g

r a

d

at

i

o n

i

s also

i

n

f l

u e n c e

d b y

s uc

h

t

h

in

g

s a s te

m p

e r atu r e, p H , n utrie nts, o xy ge n, a n

d

so

i l

ty

p

e.

PA H

s c ons

i

s t o

f 2

-

7 b

en ze ne r

i

n

g

s ;

PA H

s w

i

t

h l

es s t

h

a n

f

ou r

b

e nz e n e r

i

n

g

s ma

y

b

e v o

l

at

i l i l i

ze

d

,

p

hotoox

i d i

ze

d

, ox

i d i

ze

d

,

b i

oa ccu

m

u

l

at e

d

, a

b

so r

b

e

d i

n t

h

e s o

i l

,

l

e ac

h

e

d

or

b i

o

d

e

g

r a

d

e

d b y m i

cr o o r

g

an

i

s

m

s

(

C

e r n

i g l i

a,

19 9 2

;

P

ar

k

, e t a

l

,

19 9 0

;

B

o s s ert a n

d

B

a r t

h

a,

1 9 8 6

)

.

P A H

s w

i

t

h f

ou r or

m

ore

b

enz e n e r

i

n

g

s are

l

e s s

b i

o

d

e

g

ra

d

a

b l

e,

l

e s s

m

o

b i l

e , a n

d p

e r s

i

st

l

on

g

er

i

n en v

i

ron

m

ent a

l m

a tr

i

c es.

H i g h m

o

l

e c u

l

ar we

i g h

t

P A H

s

ar e

d

es or

b

e

d

s

l

ow

l y

an

d

t

h

e re

f

o re a re

l

es s a v a

i l

a

b l

e

f

or

b i

o

l

o

g i

ca

l

u

p

t a

k

e .

T h

e

h

a

l f

-l i

v es o

f h i g h m

o

l

e c u

l

a r we

i g h

t

P A H

s

l i k

e

b

en z o

[

a

]

p y

r e n e a re a

l

s o

l

on

g

e r t

h

an

l

ow

m

o

l

e cu

l

ar

w

e

i g h

t

PA H

s

l i k

e n a

p h

t

h

a

l

e ne.

T h

e se

P A H

s ar e rec a

l

c

i

tr a nt a n

d

c a n

p

e rs

i

st

i

n t

h

e en v

i

r o n

m

e nt

f

o r

l

on

g p

er

i

o

d

s o

f

t

i m

e

(

C

er n

i g l i

a,

19 9 2

)

.

R

e c e n t

l y

, r e s e a r c

h

h

a s

b

e e n c o n

d

uc t e

d

t o te s t t

h

e

d

e

g

ra

d

a

b i l i

t

y

o

f

h i g h

er

m

o

l

ec u

l

ar we

i g h

t

P A H

s.

A

stu

d

y

b y

M

a c

G i l l i

v r

y

an

d Sh i

ar

i

s

i

n

19 9 3 f

ou n

d y

ea st t o

h

a v e t

h

e a

b i l i

ty t o

d

e

g

r a

d

e

p h

ena n t

h

r e ne a n

d b

e n z o

{

a

}

a nt

h

r a c e ne

(

M

ac

G i l l i

v e ry a n

d

Sh i

a r

i

s,

19 9 3

)

. It

h

a s a

l

so

b

e e n

f

o u n

d

t

h

a t

M

y

co

b

a c t

i

u

m

s

p

.

PY R

-

1 h

a s t

h

e a

b i l i

ty to

m i

n e r a

l i

ze

f l

u o rant

h

en e, n a

p h

t

h

a

l

e ne , a n

d p y

r ene

i

n

p

ure cu

l

tur e a n

d

a

l

s o e n

h

a n c e

m i

n e r a

l i

za t

i

on o

f

f l

uorant

h

ene a n

d

p

yr e ne

i

n se

d i m

e nts c o nta

i

n

i

n

g

n at

i

v e

(9)

T he t ox ic,

m

utage nic, a nd cci r c

in o

ge n ic n atu re of P A H s is a c o n c ern to both

h

u

m

an a n

d

e n vir o n

m

enta

l he a

l

t

h

(

A g

e n c

y f

o r

T

o x

i

c

S

u

b

st anc e a n

d D i

s e a se

Re

g i

str

y

,

19 9 3

)

.

E

x

p

o s u re to

PA H

s

p

r o v es t o

b

e a s

i g

n

i f

ic ant

h

e a

l

t

h

r

i

s

k

t o

p

e o

p l

e

l iv

i

n

g i

n t

h

e

i

n

d

u str

ia

l

iz e

d

a r ea s o

f

t

he

wo r

l d

. For ex a

m p l

e ,

P

A

H

c o ntam

i

n at

i

on r a n

g

e s

f

ro

m 5

n

g / g

o

f

so

i l in

u n

d

ev elope

d

a r eas an

d

1

.7

9

x

10

ng

/ g

at a n o

i l

r e

f i

n e ry

(

C

e r ni

g l

ia,

19 9 2

)

.

T h

e

17 7 5

s tu

d

y

b y P

o t t

f

ou n

d

c

h i m

ney sw ee

p

s to

h

a v e a

h ig h i

nc

i d

enc e o

f

c a n c er ,

p

r e s u

m

a

b l

y

d

u e t o t

h

e

i

r e x

p

os u r e t o P

A

H s

(

A g

enc

y f

o r

T

o x

i

c

S

u

b

sta n c e a n

d D i

s e a s e

R

e

g i

s try ,

19 9 3

)

.

I

n t

h

e

19 3 0

'

s

B

e n z o

{

a

}

p y

r e n e was

is

o

l

at e

d

f

ro

m

c oa

l

t ar , a

cons t

i

tuent

f

ou n

d in

soot, a n

d f

ou n

d

t o

b

e c ar c

in

o

g

e n

i

c .

C

on ce n tr at

i

ons o

f P A H

s

in

coa

l

t ar c an

b

e or

d

e rs o

f m

a

g

n

i

tu

d

e

g

r e ate r t

h

a n t

h

ose

f

o u n

d i

n

p

etr o

l

e u

m d

e r

iv e

d

m

at er

i

a

l

s

(

M

ac

h

a

d

o e t a

l

,

19 9 3

)

.

P

A H

e x

p

osu r e

h

a s no t on

l y b

e en

l i

n

k

e

d

to s e v e ra

l

ty

p

e s o

f

can c e r

b

ut

i

t

i

s a

l

s o

i

n vo

lv

e

d i

n a

f f

e c t

i

n

g

t

h

e

h

e

m

a t o

p

oe t

i

c an

d

t

h

e

i m m

u n e s

y

st em . P

A H

s

m

ay a

f f

ect t

h

e

h

u

m

a n

f

e tu s as we

l l

(

A

g

en c

y f

or

T

ox

i

c

S

u

b

stan c e a n

d

D

i

sea s e

R

e

g i

str

y

, 1

9 9 3

)

I

n

19 9 3

,

A

s

p

o sto

l i

et a

l

. s tu

d i

e

d

t

h

e e

f f

e c t s o n wor

k

e rs o

f p

ro

l

on

g

e

d

ex

p

o s u r e t o c ut t

i

n

g

f l

u

id

s ; cut t

i

n

g f l

u

i d

s a r e

m ix

tur e s conta

i

n

in

g

var

i

o u s

p

er c ent a

g

es o

f m i

n er a

l

o

i l

s an

d /

or ,

m

or e r a re

l

y , synt

h

et

ic

o

i l

s.

T h

es e

f l

u

id

s ar e u se

d m

a

in

l y i

n me c

h

an

i

ca

l

en

g i

ne er

in

g

an

d

t

h

e

m

et a

l f

a

b

r

i

cat

i

o n

i

n

d

us tr

y

.

I

t w a s c on c

l

u

d

e

d f

r o

m

t

h i

s re s ea rc

h

t

h

a t

P A H

s

w

e r e

m

a

i

n

l y

re s

p

ons

i b l

e

f

or c ar c

in o

g

e n

i

c act

iv

i

t

y

(

s

k i

n an

d

re s

p i

rator

y

t u

m

or s

)

a ssoc

i

a t e

d

w

i

t

h

t

h

e u se o

f

cut t

i

n

g fl

u

id

ov e r t

i m

e .

P A H

s,

m

or eover , are

l i

ste

d

o n t

h

e

(10)

II P

R

e

g i

stry ,

19 9 3

)

.

A

p

p

r opr

i

ate e v a

l

u at

i

o n o

f p

o te nt

i

al a

d

v e r s e r

i

sk

f

r o

m

P A H e x

p

o s u r e

i

s

d

e

p

e n

d

ent o n t

h

e a

p p

r opr

i

ate to x

i

c o

l

o

g i

c a

l

a ss ess

m

e n t o

f PA H

s.

T

ox

i

c

i

t

y

asse s sme n ts u s

i

n

g

ex

p

er

i m

e n t a

l

an

i m

a

l

s

h

ave

b

ee n

p

er

f

orme

d

f

or

i

n

d i

v

i d

ua

l P A H

s

(

K

w an ,

D

ut

k

a, a n

d L i

u ,

19 9 0

)

. In t

h

e e n v

i

ronme nt,

h

owe v e r,

PA H

s

ar e a

l m

o st al ways

f

o u n

d

as c o

m p

o u n

d m i

xtu res.

M

i

x tu r e s exist

i

n t

h

e se

d im

e nt

b

e c a u s e

s e

d i m

e n t c ont a

m i

n a t

i

o n usu a

l l y i

n vo

l

ve s t

h

e s et t

l i

n

g

o

f m

any c

h

em

i

c a

l

s

(

S

war tz e t a

l

,

19 95

)

.

I

t

h

as

b

e e n

f

ou n

d

t

h

a t

P

A

H m i

xtu re s ar e so

m

et

i m

e s

m

or e to x

i

c t

h

a n

i

n

d i

v

id

u a

l

P A H

t ox

i

c

i

t

y

.

T h

e co

m b i

ne

d

s tr e ss es o

f m

u

l

t

i p l

e cont a

m i

n a nts, o r

i

n t

h i

s case

PA H

s,

m

ay

ca u s e e nv

i

r on

m

ent a

l

e

f f

ec ts t

h

a t wou

l d

no t

b

e e x

p

e cte

d

w

h

en

PA H

s a re tes te

d i

n

d i

v

id

ua

l l y

.

P

A H m i

xtu r es a re

m

ore res

i

stant t o

d

e

g

r a

d

at

i

o n a n

d

e x

h ib i

t a nta

g

o n

i

st

i

c, syn e r

g i

st

i

c, o r

a

dd i

t

i

v e c

h

a r a cte r

i

st

i

c s

(

C h

au

d

r

y

,

199 5

;

K

e

l l

e

y

a n

d C

er n

i g l i

a,

199 5

)

.

M

i

x tu r e s t

h

at e x

h ib i

t ant a

g

o n

i

st

i

c c

h

ar a c te r

i

st

i

cs are t

h

o s e t

h

a t s

h

ow a

l

e s s t ox

i

c

re s

p

o n s e t

h

a n t

h

e su

m

o

f

t

h

e

i

n

d i

v

i d

u a

l

c o

m p

ou n

d

s t e ste

d

.

Sy

n e r

g i

s t

i

c r es

p

o n se s w

i l l

o c cu r

w h

en t

h

e

m i

xt ure to x

i

c

i

t

y i

s

g

r ea t er t

h

an t

h

at o

f

t

h

e

i

n

d i

v

id

ua

l

c o

m p

oun

d

s te s te

d

.

A dd i

t

i

ve

t ox

i

c

i

t

y

r e s

p

o ns e s are t

h

ose t

h

at

h

a v e

m i

xt ur e t ox

i

c

i

ty e

q

u a

l

to t

h

e s um o

f

t

h

e

i

n

d i

v

i d

u a

l

co

m p

ou n

d

to x

i

c

i

ty.

A

1990

s tu

d

y w

i

t

h b

en zo

(

a

)p y

ren e

(

B A P

)

showe

d

t

h

a t s

y

ne r

g i

s t

i

c

to x

i

c

i

ty res

p

o n ses oc cu r r e

d

w

h

en B

A P

wa s te st e

d i

n a

b i

nar

y m i

xtu r e o

f p

o

l y

c

h

lor ina te

d

ar o

m

a t

i

c

h y d

r o c ar

b

ons

(

D

ut

k

a an

d K

w a n,

19 82

a n

d D

o n n e

l l y

a t a

l

,

199 0

)

,

X

P

A

H

M

o

d

e

l

T h

e

S PA H

M

o

d

e

l

wa s

d

ev e

l

o

p

e

d i

n

19 95 by D

r.

R

o

b

ert

S

wartz to e s t

i m

at e t

h

e

t ox

i

c

i

t

y

o

f P A H

(11)

c o

m p

ou n

d d

ata.

B

e

f

o r e th is, c o nta

m

in ated P A H site s we r e e v alu ate

d b

a s e

d

on in

d

iv

id

u al

P A H

to x

i

c

i

ty.

T h

e

S P A H m

o

d

e

l

assu

m

e s t

h

at

f

iel

d

co

l

le c t e

d m i

xtur e s ex e rt a t ox

i

c str ess

t

h

at

is

d

ue t o t

h

e a

d d i

t

iv e t o

x

i

c

i

t

y

o

f

t

h

e

i

n

d i

v

id

ua

l

co

m p

ou n

d

s

p

r e sent.

I

t

is th

ere

f

or e

p

ossi b le t o

p

r e

d i

ct P

A H

to x

i

cit

y b

y calculat in

g

the t ox ic u n

i

ts of the

i

n

d i

vi du al c om po u n

d

s,

i

n th is case , u s

i

n

g

a

m p h ip

o

d da ta

.

T

ox

i

c un

it

s

(

T

U

)

were ca

l

c u

la t

e

d b

e cause t

h

e

y

can

b

e

de

r

iv ed f

rom a

l l

t

y p

e s o

f

to x

i

c

i

ty

m

e a s u r e

m

e nts a n

d

correc te

d f

o r the so

l

u

b i

l

i

t

i

e s o

f

the c o

m p

ou n

d

.

T h

e

T U is

b

ase

d

o n t

h

e s o

l

u

b i l i

t

y l

im it s o

f

t

h

e

P A H

s

in

t

h

e a

d d it

i

v

it

y m

o

d

e

l

.

I

t can

b

e

d

e

fi

n e

d

a s t

h

e u n

i

t

e

q

u a

l

t o t

he c o n c e

nt rat

io n

o

f

a c

he

m i

c a

l

tha t

k il

ls

5 0

%

o

f

tes t s

p

e c

i m

ens

i

n a t ox

ic

it

y t e s t

(Sp

ro u

g

e,

19 70

)

.

T U

wer e ca

l

c u

la t

e

d b y m

u

lt

ip l y i

n

g

t

he

P A H

wa te r s o

l

u

b i l i

t

y b y

t

h

e

L C

5 0

o

f

t

h

e

PA H f

or e ac

h in

d i

v

id

ua

l

c o

m p

ou n

d

.

T

o

d

a t e ,

13 PA H

s

h

ave

b

ee n t e s te

d

w

it

h

p

ro

m i

s

i

n

g

r e s u

lt

s.

T h

e tota

l

n u

m b

e r o

f

t o x

ic

u n

i

ts o

f

the

PA H

s

(

I T U

)

wa s c a

lc u

l

a t e

d

,

assu

m i

n

g

a

d d it

i

v

i

t

y

,

b

y a

d d i

n

g

t

h

e

i

n

d i

v

i d

ua

l T U

s o

f

e a c

h

c o

m p

o u n

d

(

S

wartz et a

l

,

19 95

)

.

T h

e

I P A H m

o

d

e

l

was ver

i f

i e

d b y

co

m p

a r

i

n

g m

orta

l i

t

y p

r e

d i

ct ed by the

m

o

d

e

l

w

i

t

h

t

h

e

m

ort a

l it

y

o

b

s erv ed in s e

d i m

e n t t ox

ic

i

t

y

te sts

i

n

fi

e

ld

-c o

l l

e c t e

d

s a

m p l

e s.

I

t wa s

f

ou n

d

t

h

at t

h

e

m

o

d

e

l

acc ur a te

l y p

r ed ic t e

d

the t ox

ic

it

y

o

f PA H

-c onta

m i

n a te

d

s e

d i m

ent s w

h

en

P A H

s we re t

h

e

p

r

i

nc

i p

a

l

c ont a

m i

n ant.

H

ow e v e r

, w

h

en a

PA H

wa s not t

h

e

p

r

i

nc

i p

al

c o n t a

m i

n ant, s

ig

n

i fi

c a n t

d i ff

eren ce s

b

e twe en t

h

e

p

r e

d i

c

it e

d

an

d

t

h

e o

b

ser v e

d

to x

ic

it

y

wer e

o

b

se rve

d

(

S

war tz e t a

l

, 1

9 95

)

.

T h

ere

i

s an

86

.

6

%

c orr e s

p

on

d

ence an

d

no s

i g

n

if

ica nt

d if f

er ence

b

e twe e n

p

r e

d i

cte

d

to x

i

c

it

y f

r o

m

t

h

e

m

o

d

e

l

an

d

a c tua

l

o

b

ser ve

d

to x

i

c

i

t

y

at

P A H

co ntam

i

nate

d

s

it

es w

he

re

(12)

M^ s e

d i m

e nt qu al ity gu

i d

e

l

in e s s u c

h

a s :

E P A S

e

d i m

e nt

Q

u al ity

C

rite ria

(

E

P A

A S

Q

C

)

f

o r t

h

e

p

r ot e c t

io

n o

f b

enth ic or

g

an

is

m

s.

A pp

arent

E f f

e ct

T h

re sho

l d

(

A

E T

)

,

St

at e o

f

W

a s

h i

n

g

t on

C l

ean u

p S

cr een

i

n

g L

eve

l

s

( W

A C S L

)

,

S

t a t e o

f

W

as

h i

n

g

t on

S

e

d i m

e nt

Q

ua

l i

t

y S

ta n

d

a r

d

s

(

W

A S

Q

S

)

,

E ff

e ct s r a n

g

e

-

L

ow

(

E R L

)

,

E f f

ec t s ran

g

e

-h i g -h

(

E F H

)

, an

d Sc

re en

i

n

g L

eve

l

C

o n c e ntr at

i

o n s

(

S

L

C

) (

S

wa rtz et al,

19 9

5

)

.

T h i

s a

p p

ro a c

h i

s st

i l l b

e

i

n

g

stu

d i

e

d

as a wa

y

t o

p

r e

d ic t

P A H m i

xtu re t ox

i

c

it

y i

n

s e

d i m

e nt, a n

d

t

h

e r e a r e so

m

e u n c e rta

i

nt

y

a n

d l i m i

tat

i

o n s to

it

.

F

o r e x a

m p l

e

, t

h i

s mo

d

e

l

d

o e s n ot ta

k

e

i

nt o a c c ou nt t

he

un c ert a

i

nt

ie s

b

e twe en

m

ar

i

ne an

d

e s t u ar

i

ne a

m p h ip

o

d

s

in

P A

H

se n s

it

i

v

it

y.

T h

e S P A

H

M

o

d

e

l

a

l

so

do e

s n ot a c o u nt

f

o r t

h

e

p

ot ent

i

a

l

e

f f

e cts o

f

n atu r a

l

v a r

i

a

b l

e s s u c

h

a s se

d im

e nt

g

r a

i

n s

i

z e, tota

l

o r

g

a n

i

c c a r

b

o n, a n

d

r e

d

o x

p

ote nt

i

a

l

(

S

wartz et a

l

,

19 95

)

.

M

i

cr oto x

"

-Ac u t e

Si

nc e

i

t

'

s

d

eve

l

o

p

me nt

i

n

19 7 9 b

y

B

e c

k m

a n

I

n str u

m

e nt s, t

h

e

M

i

c ro t ox

* a s s ay

h

a s

b

een us e

d

t o

d

e t er m

i

ne an

d m

on

i

t or s o

i l

a n

d

s e

d i m

ent, wa s t ew at er , su r

f

a c e wa te r ,

a n

d

a

l

s o

g

ro u n

d

wate r t ox

i

c

it

y

(

B

u

l ic

h

,

19 8

4 ;

S

o

m

a su n

d

o r a

m

et a

i

,

19 90

)

.

S

o

m

e s t a te ,

an

d

f

e

d

er a

l

re

g

u

l

at or

y

a

g

enc

ie s

us e t

h i

s s

y

ste

m

i

n s c r e e n

i

n

g

t e s t s to

m

o n

it o

r

en v

ir o

n

m

ent a

l p

o

l l

ut ant s

(

So

m

a su n

d

o r a

m

e t a

l

,

19 9 0

)

.

M

ic

ro t ox *

is

us e

d b y

t

h

e

C i

t

y

o

f C h

at ta n o o

g

a t o r a n

k

a

l l

the

l

o c a

l i

n

d

u str

i

a

l d i

s c

h

a r

g

e s

i

n o r

d

e r o

f

r e

l

at

i

v e t ox

i

c

it

y

an

d i

n or

d

er o

f

re

la t

i

ve to x

i

c

i m p

a c t on t

h

e

C i

t

y

'

s wa s t ewa t er tr e a t

m

ent

p l

ant

(

K

u r z,

19 84

)

.

M

i

c ro t ox *

h

a s

b

e en r e c o

m m

en

d

e

d b y T h

e

A m

e r

i

c a n

S

o c

ie t

y f

o r

T

e s t

i

n

g

(13)

^

M

ic r ot ox *

f

or te st

i

ng tr e ate

d

e

f f l

u e nt s,

f

o r

p

re

d i

c t

i

n

g

lan

d

tr e at a

b i l i

t

y

o f or

g

a n ic

w

a ste s

an

d f

or

b i

oa s se s sme nt o

f

w a s t e

d i

s

p

o s a

l

s

i

t e s

(

M

u n

k i

t tr

i

c

k

e t a

l

,

19 9 1

)

.

T

a

i

w an

h

a s

b

e

g

u n t o u s e t

h

e

M

i

c r o t ox * s

y

s te

m

to s cree n

i

ts

i

n

d

us tr

i

a

l

was t ewat ers

(

H

a o et a

l

,

19 9 6

)

.

M

i

cr oto x

^

i

s a t ox

i

c

i

t

y

test t

h

at

h

a s

b

e c o

m

e o ne o

f

t

h

e s tan

d

ar

d

t ox

i

c

i

t

y

a s s a

y

s

(

T a rk

p

e a et al, 1

9 8 6

)

.

M

i

cro t ox

^

m

ea su res t

h

e

d

e c rea s e

d

b i

o

l

u

m i

n e s c e nce o

f

t

h

e

b

a cte r

i

a

Vi b

r io

f

i

s c

h

er

i i

n t

h

e

p

re s e n c e o

f

a t ox

i

c a

g

ent.

T h

e

l

u c

i f

e r ase e n z

y

me

f

r om t

h i

s

b

ac te r

i

a

u ses re

d

u ce

d f l

av

i

n a n

d

o xy

g

en to

p

r o

d

u c e

l i g h

t

(

C h

an

g

et a

l

, 1

9 8 1

)

.

A p h

ot o

m

et er

m

e asu r e s t

h

e

d

e c rea s e o

f l i g h

t a n

d g i

ves a r e a

d i

n

g

o

f

t

h

e

p

er c e nt

l ig h

t

l

o ss,

(

G

a

m m

a

)

.

A

n

E f f

e ct

C

o n centr at

i

o n o r

E C

50

i

s t

h

e n

p

r o

d

uce

d b

y

g

r a

p h i

n

g

t

h

e

l

o

g

o

f

t

h

e

G

a

m m

a

r es

p

on se ver su s t

h

e

l

o

g

o

f

t

h

e s a

m p l

e con centr at

i

on .

T h

e

E C

5 0

i

s t

h

e c

h

e

m i

c a

l

o r

e nv

i

r on

m

e n t a

l

s a

m p l

e c onc entr a t

i

o n ca us

i

n

g

a

5 0

%

d

e cr e as e

i

n t

h

e

M

i

c r o tox *

R

e a

g

e n t

l i g h

t out

p

u t u n

d

er

d

e

fi

ne

d

co n

d i

t

i

o ns o

f

t

i m

e a n

d

te st tem

p

er atu r e

(

M

i

c ro

b i

cs,

19 8 9

)

.

T h

e

d

e cr ea se

d i

n t en s

i

t

y

o

f b i

o

l

u

m i

ne s ce nce

i

s

p

r o

p

o r t

i

o n a

l

to t

h

e to x

i

c

i

ty o

f

s

p

ec

i f i

c

c

h

e

m i

c a

l

s or e n v

i

r o nme nt a

l

s am

p l

es.

T h

e s

m

a

l l

e r t

h

e

E C

5 0

,

t

h

e

m

o re t ox

i

c t

h

e

c

h

e

m i

c a

l

o r e n v

i

ron

m

e nt a

l

s a

m p l

e .

R

ese ar c

h h

a s s

h

own t

h

a t t

h

e r e su

l

ts o

f

M

i

cro t ox *

are co

m p

ara

b l

e t o o t

h

er

a s sa

y

s.

C

a n na

-

M

i

c

h

a e

l i d

ou

i

n

19 9 3

, c o

m p

are

d

M

i

cro t ox *

t o a

g

a s c

h

ro

m

a t o

g

ra

p h

y

"

fi

n

g

er

p

r

i

n t sc re en

i

n

g

t e s t" .

H

e c o

m p

a re

d

the to x

i

c e

f f

ects o

f 7 m i

xtu r es o

f

vo

l

at

i l

e

o r

g

an

i

c

p

o

l l

ut ants

(

V O C

)

. In

d

o in

g

so,

h

e

f

ou n

d

t

h

at the

M

i

c r ot ox

*

(14)

-m

2 2 0 ti

m

e s

m

o r e s e n sitiv e t

h

a n the

G C /

F

I

D fo r a r o

m

atic

m

ix tu r e s. T h is te st wa s als o

f

ou n

d m

or e s e n s

i

t

iv e

fo

r

d

etec t

i

n

g

r e

g

u

l

a r

p

etro

l

eu

m

an

d d ie s e

l

t ox

ic

i

ty.

R

e s e ar c

h h

a s a

l

s o

b

e en

d

o ne c o

m p

ar

i

n

g

M

ic r

o to x* to

f

ish,

D

a

p h

n

i

a , an

d

othe r

m ic r

o

b i

a

l

t ox

i

c

i

ty

m

et

h

o

d

s.

M

ic r oto x "

v s. F ish. T he

2

4

-to -

9 6

-

h

r f is

h

Lethal

C

o n c e ntr ation 5

0

(

L

C

5 0

)

acute

b io a

s sa

y i

s one o

f

t

h

e

m

o st w

i d

e

l

y use

d

a cu te tox

i

c

i

t y t e s ts.

T h

e re

h

a s

b

e en ext ens

iv

e

re se a r c

h

s

h

ow

i

n

g

t

h

at

M

ic r

o tox *

is

a s sens

it

iv

e a s t

h i

s tes t, e s

p

e c

i

a

l l

y w

h

en us

i

n

g

t

h

e

f la t

h

e a

d m i

nnow

(

In

d

ora to et a

l

,

C

u rt

i

s et a

l

,

19 82

,

E

.

V

.

S

.

C

onsu

l

t ants

,

19 8 9

,

B

u

l i

c

h

e t a

l

,

19 80

,

M

un

k it t

r

ic

k

e t a

l

,

19 9 1

)

.

I

n

19 9 3

,

D

e

Z

wa rt an

d

S l

o o

f

d

ete r

m in

e

d

the t ox

ic

i

t

y

o

f

15

c

h

e

m i

c a

ls

b y

co

m p

a r

i

n

g

the

M

ic r

o to x *

a c ute a ss a

y

to that o

f g

ree n a

l g

ae,

D

a

p h

n

ia

, a n

d fi

s

h

.

T h

e

ir

r e su

l

t s sho we

d

t

h

e

M

ic

r ot ox a s sa

y

t o

b

e c o

m p

ara

b le

to the o ther t e s ts, e s

p

ec

i

a

l l y

t

h

e

acute t ests c on

d

uc te

d

w

i

t

h fi

s

h

.

D i f f

e r e nc e s

b

e twe en t

h

e

fi

s

h

a n

d

M

i

cr o t ox

®

s ens

i

t

iv

i

ty

w

ere w

i

th in one o r

d

er o

f

ma

g

n

i

tu

d

e .

T h

e

C i

t

y

o

f C h

at t a n oo

g

a

h

a s n ot on

l y f

ou n

d

t

h

e t e s t t o

be

m

o re se ns

i

t

iv

e,

b

ut

a

ls o

l

e s s ex

p

ensiv e an

d

t

i m

e c onsu

m i

n

g

.

C

o

m p

ar e

d

t o t

h

e

S

tate o

f T

e n n ess ee

, w

h i

c

h

a v e r a

g

e

d 10

o r

le s s

b io

a ssays a

y

e ar us

i

n

g

the L

C

5 0

fi

s

h

t ox

i

c

it

y

te st,

C h

a t ta n oo

g

a

c on

d

uc te

d 160

t e s t s us

i

n

g

M

ic r

otox*

in

l

e ss t

ha n

6 m

ont

h

s t

i m

e

(

K

u r z,

19 8 4

)

.

M

i

c r otox'^ vs.

D

a

p h

n

ia

.

T h

e c o

m p

ara t

iv

e s e n s

it

iv

it

y

o

f

M

ic

ro t ox

® an

d

D

a

p h

n

i

a

is

d

e

p

e n

d

e nt o n t

h

e c o

m p

ou n

d

t e s t e

d

.

M

ic r

o t ox

®

i

s c o

m p

a r a

b l

e to

D

a

p h

n

ia

(15)

e x a

m

p le ,

D

a

p

hnia is

m

o r e s e nsitiv e t o a

m m

onia, cya nide , he x a ch lo r o e thi a n e ,

p

e n ta

d i

one, an

d

s o

d i

u

m i

au r

y l

.

M

i

cro t o x *

, o n t

h

e o t

h

er

ha n

d is

m

o re se ns

i

t

iv

e to

c

h l

oro

f

o r

m

, styr en e a n

d h i g h l y

s u

b

st

i

tut e

d

or

g

a n

ic s

(

M

u n

k i

t tr

i

c

k

et a

l

,

19 9

1

)

.

M

i

c r o to x "

vs. o the r

M

i

c ro

b ia

l

to x

ic

i

t

y m

e t

h

o

d

s. In 19 8 1, D ut

k

a a n

d K

wan

c o

m p

a re

d

M

i

c r ot ox* t o thr ee othe r

m

icr ob ia

l

to x

ic

ity te st s:

Sp

iri

ll

u

m

v o

l

uta n s,

P

seu

d

o

m

a s

f

lu

o re s c e n s, an

d A

ero

m

ona s

hy d

ro

p h i l

a .

T he

M

ic r o

t ox *

te st

p

r o ve

d

t o

b

e

t

h

e

m

o s t s ens

i

t

iv

e t o t

h

e c

h

e

m i

ca

l

s tes te

d

(

w

i

t

h

t

h

e e x c e

p

t

i

o n o

f H g

'

^' *"

,

N i

"

^" ^

,

P b

"

^ "*"

,

an

d N

-

N i

tr o s o

d i

e t

h y l

a

m i

ne

)

.

M

ic r

o t ox ®

wa s

p

rov e n to

b

e a

m

o r e r e

l i

a

b l

e a n

d

s ens

i

t

iv

e

te s t

w h

e n co

m p

a re

d

t o t

h

e r es az u r

i

n r e

d

uct

io

n

(

R R

)

m

et

h

o

d

an

d

t

h

e

d i

sso

lv

e

d

oxy

g

en

d

e

p l

e t

i

on

(

D O D

)

t e s t

(

G

r een e e t a

l

,

19 85

)

.

M

i

cro t ox *

a

l

s o ta

k

e s

l

e s s t

i m

e t

h

an o t

h

e r

b i

oa ss a

y

s an

d h

a s a s t a t

i

s t

ic

a

l

a

d

vant a

g

e s

in

c e r esu

l

ts ar e

b

a se

d

o n us

i

n

g

over

10

^

b

acte r

ia

.

W

h

en

M

c

F

et er s t e s t e

d

t

h

e two or

g

an

i

s

m

T

c

h

an

p

ro c e

d

u r e

(

a

l g

ae an

d

lu

m i

nes c e nt

b

ac te r

i

a

)

w

i

t

h

M

i

c r oto x *

h

e

f

o u n

d

M

ic r o

t ox

'

t o

b

e a

m

ore s ens

i

t

iv

e te s t .

R

e

l

ativ e sens

i

t

iv

i

ty t o

M

i

cr ot ox *

h

a s

b

e en c om

p

are

d

t o a va r

i

e ty o

f b i

o a s s a

y

m

e t

h

o

d

s

f

or t

h

e tox

i

c

i

t

y

o

f

c

h

e

m i

ca

l

s, s e

d i m

ent s, an

d

co

m p l

ex e

f f l

uent s.

W

h

en

c o

m p

are

d

t o

f i

s

h

an

d D

a

p h

n

i

a stu

d i

e s, s ever a

l

r e s e arc

h

e r s r eac

h

e

d m

a n

y

o

f

t

h

e s a

m

e

m

a

j

o r c onc

l

u s

i

ons.

M

u n

k i

t tr

i

c

k

s s u

m m

a r

i

z e s t

h

e s e con c

lu s

io n s

as

f

o

l l

ow s

(

M

u n

k i

t tr

i

c

k

e t a

l

,

19 9 1

)

:

O

r

g

a n

ic

s -

M

i

cro to x *

is

as s e ns

i

t

iv e

a s ot

h

er a s sa

y

s

f

o r t

h

e

m

eas u re

m

ent o

f p

u r e

o r

g

a n

ic

s.

M

ic r

o to x *

(16)

4

»

^>>Vi|

m

ult

i

ch lo r

i

n ate

d b

e n z e n e s, p

h

e n o

l

s, a n

d

et

h

ano

l

s.

T h

e sys te

m i

s

l

es s s en s

i

t

i

v e t o

c

y

an

i d

e, c

h l

oro

f

o r

m

, o r

p h

e n o

l

.

In o r

g

a n

i

cs -

M

i

cr oto x *

i

s n ot a s s e n s

i

t

i

v e a s ot

h

er te st s to

m

o s t

i

n o r

g

a n

i

c s . It

i

s

h

owever ,

m

ore se ns

i

t

i

ve t o

m

ercu r

y

, ars en

i

c , a n

d

c o

b

a

l

t.

M

u n

i

ci

p

a

l

W

as te s -

M

i

c ro to x

*

i

s a

f

av o r a

b l

e

b i

oa s s ay

f

o r mo n

i

to r

i

n

g m

u n

i

c

i p

a

l

was tes.

In

d

u str

i

a

l

W

a stes -

M

i

c ro to x® c a n a

l

s o

b

e use

d

t o s c ree n

f

o r t

h

e r e

l

at

i

v e to x

i

c

i

t

y

o

f

c o

m p l

ex e

f f l

u e nts.

S

e

d i m

e nt

C

ont a

m i

na t

i

o n -

T h

e s e ns

i

t

i

v

i

t

y

o

f

M

i

cro to x *

va r

i

e s w

i

t

h

c ont a

m i

n a nt

e x tra c t

i

o n te c

h

n

i q

u e

(

M

u n

k i

t tr

i

c

k

et a

l

,

1 9 9 1

)

.

M

i

cro t ox ®

no t o n

l y h

a s

b

e e n u s e

d

to te st

i

n

d i

v

i d

ua

l

to x

i

ca nt s,

b

u t

h

as a

l

s o

b

e e n

u s e

d

to te st s o

m

e

m i

xtu res.

M

a n

y h

ave

f

ou n

d m i

x tu r e s t o

b

e the s u

m

o

f

t

h

e

i

r t ox

i

c

p

art s or a s

b

e

i

n

g

a

dd i

t

i

ve

(

S

e

l l

er s an

d R

am

)

.

H

ow ever

,

i

t

h

a s

b

e en

d i

s c o v ere

d

ov er

t

i m

e t

h

a t

m i

xtu r e s can r eact syn e r

g i

st

i

ca

l l y

a n

d

anta

g

o n

i

st

i

ca

l l

y a s we

l l

.

In

C

an na-

M

i

c

h

a e

l i d

ou '

s stu

d y

o

f

v o

l

a t

i l

e or

g

an

i

c

p

o

l l

uta nts,

h

e

f

oun

d

m i

xtu re s t o s

h

ow a

d d i

t

i

ve e

ff

e c ts

(

C

an n a-

M

i

c

h

a e

l i d

ou ,

19 9 3

)

.

B

a s e

d

o n ex

p

o su re

t

i m

e, a n

d

t

h

e

p

r o

p

or t

i

o n o

f

c

h

e

m i

ca

l

co

m p

on e nt,

h

e av

y

m

eta

l

m i

xtu r e s w er e

c a t e

g

o r

i

ze

d

a s s

y

n e r

g i

s t

i

c , anta

g

on

i

s t

i

c, or a

d d i

t

i

ve.

T h

e r esu

l

t s s

h

owe

d

t

h

a t a

f

t er a

4 5

-m i

nut e ex

p

o s u re,

C d

-

Z

n

i

n

t eracte

d

s

y

ne r

g i

s t

i

ca

l l y

,

C d

-

N i i

nt eracte

d

ant a

g

on

i

st

i

ca

l l y

,

a n

d C

o -

N i

a n

d A

s-

Pb i

nt er a c t e

d

a

d d i

t

i

ve

l

y

(

S

e

l l

er s a n

d R

a

m

)

.

W

a

l k

e r s tu

d i

e

d

t

h

e

t o x

i

c

i

t

y

o

f

z

i

n c a n

d p

e nta c

h l

o r o

p h

o

l

(

P C P

)

i

n

19 87

a n

d d

ete r

m i

n e

d

t

h

at t

h

e

i

n t e r a ct

i

o n s

b

e twe e n t

h

e tw o w e r e a

dd i

t

i

v e.

(17)

if^

m

D

utka a nd K wa n

(

1 9 8 1

)

als o te sted

m

ixtu r e s w ith

M

i

c r oto x

®

. T he

y, l i

k

e

C

an na-

M

i

c

h

a e

l i d

ou

f

ou n

d

t

h

a t

m i

xt ure s

p

ro

d

uc e

d

re s

p

o nses t

h

a t

w

e r e s

y

ne r

g i

s t

i

c ,

a nt a

g

on

i

s t

i

c, or a

dd i

t

i

v e ,

b

a s e

d

o n t

h

e co n c e ntr at

i

o n s o

f

t

h

e

i

n

d i

v

i d

u a

l

c

h

e

m i

c a

l

a n

d

t

h

e

i

ncu

b

at

i

o n o r co nta ct t

i m

e.

F

or e xa

m p l

e,

d i f f

e r e nt c on c e ntr at

i

o ns o

f p h

e n o

l

w er e

c o

m b i

n e

d

w

i

t

h

the s a

m

e c o n c e ntr at

i

o n of s o

d

iu

m l

a u ry l s ul

p h

ate.

C

o ntr o

l

s s

h

ow e

d

t

h

at

p h

e n o

l h

a

d

a

d

ec re a s e

d

t ox

i

c e

f f

e ct w

i

t

h i

ncre ase

d i

ncu

b

at

i

on t

i m

e; t

h

e t ox

i

c e

f f

e ct o

f

so

d i

u

m

l

au r

y l

su

l p h

a t e w a s s t a

b l

e ov e r a

15

-

m i

n ut e

p

e r

i

o

d

.

W

h

e n t

h

e two wer e

c o

m b i

ne

d

,

h

owe v e r, t

h

e r e wa s an

i

ncre a se

i

n t ox

i

c e

f f

ect. T

h

e

2 0 p p m p h

e no

l

co

m b i

na t

i

o n s

h

ow e

d

a n a

d d i

t

i

v e r e s

p

o n s e, w

h i l

e

2

.

5 p p

m

p h

e no

l

co

m b i

nat

i

o n w

i

t

h 0

.

5

p p m

o

f

s o

d i

u

m l

a u ry

l

su

l f

at e s

h

owe

d

a syner

g i

s t

i

c r es

p

o nse.

A

n a nt a

g

o n

i

st

i

c r e s

p

o nse

w

a s s e en w

h

en

h i g h

c o n c e ntr at

i

ons o

f H g

"^"*"

(

0

.

0 7 5 p p

m

)

we re c om

b i

n e

d

w

i

t

h Pb

^ "

^

(

5

p p m

)

.

M

i

c r o t ox

*

i

s s ens

i

t

i

ve, r a

p i d

, a n

d i

n ex

p

e ns

i

v e re

l

at

i

ve to ot

h

e r assays

(

R

ib

o

an

d K

a

i

se r ,

19 87

)

.

T h

e t a s

k

o

f b

r e e

d i

n

g

t e st or

g

a n

i

sms

i

s e

l i m i

na t e

d i

n t

h i

s s

y

s t e

m

.

A

ssa

y

s t

h

at t a

k

e two or t

h

ree

d

a

y

s u s

i

n

g

s t an

d

ar

d

t ox

i

c

i

t

y

t est c an

b

e con

d

uc t e

d i

n

f i

ve

to t

h i

r t

y m i

nu te s w

i

t

h

c o

m p

a ra

b l

e r e su

l

ts

(

D

e

Z

w art,

1 9 8 3

)

.

M

i

cr otox * -

C h

ron

i

c

T

h

e

C h

r o n

i

c t ox

i

c

i

ty t e s t

i

s a n ew a

d d i

t

i

o n to t

h

e

M

i

c r oto x *

a s sa

y

s

y

ste

m

.

T h

er e

f

o r e, t

h

e r e

h

a s n ot

b

een

m

u c

h p

u

b l i

s

h

e

d

c o nce r n

i

n

g

t

h i

s t e s t.

C h

r o n

i

c

to x

i

c

i

t

y

t e sts a r e u s e

d

t o

p

r e

d i

c t a

d

vers e

b i

o

l

o

g i

c a

l

e

f f

e c t s t

h

a t r e su

lt w

h

e n an o r

g

a n

i

s

m

i

s e xpo s e

d

o v e r

i

ts e ntir e

l if

e cyc

l

e

(

M

i

cro

b i

c s, 19 96

)

.

L i k

e t

h

e a c ute t est

, t

h

e

(18)

m

M

i

cro to x *

c

h

r o n

i

c to x

i

c

i

ty t est

h

as

b

e en s

h

own t o

b

e a s s e n s

i

t

i

v e o r

m

o r e s en s

i

t

i

v e t

h

a n

o t

h

er c

h

r on

i

c t e s t s

(

M

i

cro

b i

cs,

199 6

)

.

T h i

s t e s t a

l

so ta

k

es

l

es s t

i

me t

h

an ot

h

ers

,

2 2

h

ou r s.

T h i

s te s t u s e s t

h

e

M

o

d

e

l 5 0 0 A

na

l y

zer t o

d

ete c t

l i g h

t loss.

T

o a na

l y

ze t

h

e

r e s ults o

f

t

h

e

M

i

c r oto x ®

chr o n

i

c to xicity te st, t

h

e s o

f

twa r e syste

m T

o x

C

a

l

c

,

i

s u s e

d

.

It

i

nc

l

u

d

e s su c

h

s tat

i

st

i

ca

l

o

p

t

i

o n s a s: a

p

ar a

m

etr

i

c

h

y

p

o thes

i

s te st, a non

p

a ra

m

e tr

i

c

h y p

ot

h

e s

i

s te st, a n

d p

o

i

nt e st

i m

ate s ca n

b

e u s e

d

to

d

et er

m i

n e t

h

e

N

o

O b

ser v e

d E f f

e c t

C

o nce n tra t

i

o n

(

N O E C

)

a n

d

t

h

e

L

owe st

O b

s e r ve

d E f f

ect

C

o n c e ntra t

i

o n

(

L O E C

)

.

T h

e

N

O

E

C i

s t

h

e

h i g h

e st c o nce ntr a t

i

on te st e

d

t

h

at c au se s n o s tat

i

st

i

c a

l l

y

m

eas u ra

b l

e e

f f

ec t

o n t

h

e te st s

y

ste

m

. T

h

e L

O E C

, on t

h

e ot

h

e r

h

a n

d

,

i

s

d

i e

l

owe st conc entra t

i

o n tes t e

d

t

h

a t c aus es a stat

i

s t

i

c a

l l y m

e asu ra

b l

e e

f f

e c t o n t

h

e te st s

y

ste

m

(

M

i

c ro

b i

c s,

19 9 6

)

.

O b

j

ect

i

v es

T h i

s r e s ear c

h

wa s

d

e s

i g

ne

d

t o c o

m p

a re t

h

e t ox

i

c

i

t

y

o

f

PA H

s a s

i

n

d i

v

id

u a

l

c om

p

ou n

d

s a n

d

a s

m i

xtu r e s, a s t

h

ey ar e

f

o u n

d i

n t

h

e en v

i

r o n

m

e nt.

P A H m i

xtu r e to x

i

c

i

t

y

h

as not

b

e e n stu

d i

e

d

a s

f

u

l l y

a s t

h

at o

f i

n

d i

v

i d

ua

l

co

m p

ou n

d

s.

T h

e

M

i

c ro t ox

*

to x

i

c

i

t

y

tes t,

b

o t

h

a cute an

d

c

h

ron

i

c , wer e us e

d

t o

d

e te r

m i

n e t ox

i

c

i

t

y

o

f PA H

s an

d

t

h

e

i

nt er a ct

i

o ns o

f

t

h

e

m i

xtures.

T h

e s

p

e c

i f i

c o

b

j

ect

i

ves o

f

t

h i

s r e s ea rc

h p

r o

j

e c t w ere t o :

1

.

A

s se s s t

h

e a c u te an

d

c

h

r o n

i

c tox

i

c

i

t

y

o

f P A H

s

i

n

d i

v

i d

ua

l l y

a n

d

a s

m i

xtu r e s us

i

n

g

t

h

e

M

i

c r ot ox *

a s sa

y

s

y

s t e

m

.

(19)

^,

g

^

2

. T o dete r

m

in e i f the to x icity r e spo n s e o f P A H

m

ix tu r e s is pr ed ictab le ba s ed o n data

g

e n era t e

d

w

i

t

h i

n

d i

v

id

ua

l

c o

m p

ou n

d

s u s

i

n

g

t

h

e a

d d i

t

i

v e

m

o

d

e

l

(

S

wartz e t a

l

,

19 9 5

)

.

3

.

T

o e v a

l

u ate

d i f f

e r e n c e s

b

etwe e n acute a n

d

c

h

ro n

i

c t ox

i

c

i

ty a ssay r e s u

l

t s.

(20)

m

M

A T E R I A L S A N D

M

E T H O D S

C h

e

m i

c a

l

s

E i g h

t

P A H

s wer e eva

l

u at e

d by

t

h

e

M

i

cro t ox '*

b i

o a s s a

y

: na

p h

t

h

a

l

ene, c

h

r

y

se ne,

p y

r e n e, a nt

h

r a c e n e,

f l

u o r a nthr e n e,

f

lu o r e n e, ace n a

p h

t

h

e n e

(

A l d

r

i

c

h C h

e

m i

c a

l C

o . ,

M

il

w au

k

ee ,

W

I

)

an

d p h

ena nt

h

re ne

(

S ig m

a

C h

e

m i

c a

l C

o.,

S

t.

L

ou

i

s,

M

O

)

,

(

F i g

u re

1

)

.

P

A

H

s a

m p l

e

p

r e

p

a r a

ti

on

S

o

l

ut

i

o n s o

f i

n

d i

v

id

ua

l P A H

s wer e

p

r e

p

are

d b y d i

s s o

l

v

i

n

g

e ac

h P A H i

n t wo

s e

p

a ra te so

l

v e nts,

m

et

h

ano

l

an

d

a

2

%

N

a

C l

s o

l

ut

i

o n a t t

h

e

i

r r e s

p

ect

i

v e wa t er s o

l

u

b i l i

t

y

c onc ent rat

i

ons.

F

or

2 % N

a

C l

s o

l

ut

i

ons, satu ra te

d

s o

l

ut

i

ons o

f

ea c

h i

n

d i

v

i d

u a

l P A H w

e r e

p

r e

p

a r e

d b

y

f i

r st we

ig h i

ng t

h

e c

h

e

m i

c a

l

t o at

l

e as t

1

.

5

t

i m

es t

h

e sta n

d

ar

d

so

l

u

b i l i

ty

i

n

10

m L

o

f d i

s t

il l

e

d

,

d

e

i

o n

i

z e

d

w ate r. T

h

e s atu r at e

d N

a

C

l so

l

ut

i

o n s we r e m

i

x e

d

we

ll

a n

d

a

l l

owe

d

to s e tt

l

e .

S

u

p

ern a t ant wa s r e

m

ove

d

t o

m

a

k

e t

h

e s a

m p l

e

m i

xt ure.

S

o

l

u t

i

ons o

f

P A H

an

d m

et

h

ano

l

we re

p

r e

p

ar e

d b y d i

s so

l

v

i

n

g

t

h

e

w

e

i g h

t o

f

t

h

e c

h

em

i

c a

l

ne c es s ary to

p

rov

i d

e

w

a te r s o

l

u

b i l i

t

y

o

f

t

h

e

PA H i

n

10 m l

o

f m

et

h

a no

l

.

T

o x

i

c

i

ty c o

m p

ar

i

son s were

m

a

d

e

b

etw een

N

a

C l

s o

l

ut

i

o n s an

d m

e t

h

a n o

l

s o

l

ut

i

on s

f

or e a c

h i

n

d i

v

id

u a

l P A H

.

M

i

croto x

'

-

A

c ut

e to x

i

c

i

t y

T h

e

M

i

crotox a cut e t ox

i

c

i

t

y

te s t eva

l

uate s t ox

i

c

i

t

y b y

c o

m p

ar

i

n

g

t

h

e

l

u

m i

n e sce nce o

f

V

i b

r

i

o

fi

s c

h

e r

i

a

l

o n e to t

h

e

l

u

m i

n e s c e n c e

i

n t

h

e

p

r e sen c e o

f

a

p

a rt

i

cu

l

a r c

h

e

m i

c a

l

or e n v

i

r o n

m

enta

l

s a

m p l

e.

T h

i

s as sa

y

can

b

e c o

m p l

ete

d i

n

5

to

3

0

m

inute s.

T h

e

M

i

cr oto x

®

sys t e

m i

n c

l

u

d

e s a

M

i

c ro

b i

c s

M

o

d

e

l 5 00 A

n a

l y

zer to

m

ea s u re

Figure

Updating...

References

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