Effect of metabolism on bioconcentration of geometric isomers of d-phenothrin in fish

Chemospher¢, Vol.24, No.12, pp 2001-2007, 1992 Printed in Great Britain

EFFECT

OF M E T A B O L I S M

GEOMETRIC

MITSUGU

0045-6535/92 $5.00 + 0.00 Pergamon Press Ltd.

ISOMERS

MIYAMOTO,

ON B I O C O N C E N T R A T I O N

OF d - P H E N O T H R I N

SHOJI

SAITO*,

AND MASATOSHI

Environmental

Health

Sumitomo 4-2-I,

Takatsukasa,

YOSHIYUKI

TAKIMOTO,

MATSUO

Science

Chemical

OF

IN FISH

Laboratory,

Co.,

Takarazuka,

Ltd. Hyogo,

665 Japan

ABSTRACT

Yearling mixture

carp

(Cyprinus

of d-cis

piperonyl

evaluated.

under

factors

higher

subsequent

in e l e v a t e d

BCF's was o b s e r v e d presumably oxidative

2.3-11.

metabolism

Thus,

bioconcentration

that

in the a b s e n c e

isomer.

to a r e d u c e d

to the e l i m i n a t i o n

isomer

rate

for the d-trans

The e l e v a t i o n caused isomer.

constant

explained

butoxide

were isomer

resulted

change

observed

in

here was

by i n h i b i t e d The c o n t r i b u t i o n

of

(K3/K 2) was e s t i m a t e d

to

by a d i s t i n c t

and a s i g n i f i c a n c e

separately

isomer

but no r e m a r k a b l e

to the d-cis

the r e s u l t was well

were

for the d-cis

elimination

of

and the

isomers

of p i p e r o n y l

isomer,

(a 1:4

and the p r e s e n c e

condition

than those

in the p r e s e n c e

for the d-cis

phenomenon

test

BCF values

to 2 . 2 - f o l d

characteristic

of the d-cis

to d - p h e n o t h r i n

of the g e o m e t r i c

for the d-trans

reactions

biotransformation be

(BCF's)

by 1.1

attributable

exposed

the f l o w - t h r o u g h

exposure

BCF values

were

isomers)

It was d e m o n s t r a t e d

significantly and the

and d-trans

butoxide

bioconcentration

carpio)

oxidative

of m e t a b o l i s m

in

was e x e m p l i f i e d .

INTRODUCTION

Biotransformations rates

that

their

of x e n o b i o t i c s

bioconcentrations

in fish, are

in some cases,

significantly

2001

occur

e f f e c t e d 1,2)

at such

2002

In this

study,

d-phenothrin

to clarify

in carp,

of the c o m p o u n d

whose

the effect

bioconcentration metabolic

factors

pathway

to one a n o t h e r 3) were

separately

of p i p e r o n y l

which

butoxide

of m e t a b o l i s m

on b i o c o n c e n t r a t i o n

(BCF's)

and rates

of g e o m e t r i c

are

remarkably

measured, in the absence

is well

known

MATERIALS

AND

of

isomers

different

and the p r e s e n c e

as an i n h i b i t o r

of m i c r o s o m a l

m o n o o x y g e n a s e 4)

METHODS

Chemicals d-Phenothrin purity was

93.5%)

was used

96.3% when

(IR)-cis,trans-chrysanthemate)(chemical

(3-phenoxybenzyl as a test

determined

by HPLC

with h e x a n e / 1 , 2 - d i c h l o r o e t h a n e 1:4 when

determined

Piperonyl

butoxide,

Pure

Chemical

The optical

(Shimadzu

(Shimadzu

reagents

LC-6A,

purity

Sumipax

of IR isomer

0A-2000

The d-cis:d-trans isomer

(500/I)).

by G C - F I D other

compound.

GC-9A,

QF-I

and solvents

capillary

column ratio was

column).

were p u r c h a s e d

from Wako

Co.,Ltd.

Exposure Exposure

was c o n d u c t e d

guideline,

No.

corresponding isomers,

to 0.86

or 0 . 0 8 6 p p b prepared

was i n t r o d u c e d

exposure,

The renewal

fish and water

To clarify experiment presence

the effect

of d - p h e n o t h r i n

and

with

oil

samples

set at 14.4 were

of m e t a b o l i s m

at 0.43ppb of d - p h e n o t h r i n

of 1.0ppm of p i p e r o n y l

(Nikko

3% mean

Chemical) where lipid

times/day.

taken at the

butoxide

20 y e a r l i n g

specified

similar

8 weeks

intervals.

another

conditions

was c o n d u c t e d

as an

content 6)) were

During

on b i o c o n c e n t r a t i o n , under

test

solution,

aquarium,

20g body weight,

rate was

in the OECD

of d-cis and d-trans

3.4 or 0.34ppb caster

into a 100-L glass

(Cyprinus carpio, about

maintained.

way as d e s c r i b e d

4.3 or 0.43ppb

respectively,

emulsifier carp

305C5):

in a dynamic

for

exposure

but in the

4 weeks.

Analysis Each g e o m e t r i c QP-1000, (fragment

QF-I

was q u a n t i f i e d AW DMCS

of 3 - p h e n o x y b e n z y l

d-phenothrin) to be

isomer

on c h r o m o s o r b

for fish,

4 and 16ppb

For a n a l y s i s liquid-solid

group)

separately

column)

for water

respectively.

of the water, using

one

liter

XAD-4

and of 350

The d e t e c t i o n

for d-cis and d-trans

extraction

by GC-MF

isomers, of w a t e r

resin

(Shimadzu,

with a common m/e of 183 (parent

limits were

ion of calculated

respectively. sample

(Organo)

was

subjected

and eluted

with

to

50ml

of

2003

acetone.

The e l u a t e

was c o n c e n t r a t e d

and d i s s o l v e d

in 2ml of hexane

for

G C- M F d e t e r m i n a t i o n . For a n a l y s i s of a s o l v e n t homogenate minutes

of fish,

mixture

After

liquid-liquid

partitioning

to remove

and r e c o n s t i t u t e d The r e c o v e r i e s

respectively. throughout

rpm,

(100,

50,

of the 50ml)

the c o m b i n e d

for

hexane

was s u b j e c t e d

and a c e t o n i t r i l e

weight

10 layer

to

and open

column

eluted with hexane/dichloromethane/acetic

Cosmosile

C18 , e l u t e d w i t h m e t h a n o l / w a t e r

fat in the extracts.

in the above fortified

to 0 . 0 5 p p m

Then,

procedures

were

at 4.3 or 0 . 4 3 p p b

of d - c i s

The optical

the sample

was c o n c e n t r a t e d

found

for water

and 0 . 2 0 p p m

and g e o m e t r i c a l

to be more

ratios

90% when

and 0 . 2 5 p p m

of d - t r a n s

isomer

than

isomer)

for fish,

remained

intact

the p r o c e d u r e s .

The a n a l y s i s extraction capillary

and an a l i q u o t

concentrate

hexane

w i t h an e q u i v a l e n t

w i t h 2ml of hexane.

were

(corresponding

at 2700

The

PR,

and Nakarai

(I/1)

times w i t h h e x a n e

between

(Florisile

(100/50/1)

d-phenothrin

three

centrifugation

and c o n c e n t r a t e d .

chromatographies

(90/10))

fish was h o m o g e n i z e d

of w a t e r / m e t h a n o l

was e x t r a c t e d

each.

was dehydrated

acid

whole

for p i p e r o n y l

manner

butoxide

as d e s c r i b e d

column)

was used

in fish was also

above,

but G C - F I D

conducted

(Shimadzu,

w i t h a same

GC-14A,

DB-5

for q u a n t i f i c a t i o n s .

RESULTS

Average compound

actual in w a t e r

3 . 4 p p b or 4.3ppb or 0.43ppb, Fish

concentrations were

0.72

no symtoms of each

a ft e r

exposure,

2 weeks

The o b s e r v e d

in Table

isomer

by 1.1

pairs)

reveals

as shown showed

isomers

of the test

concentrations

and 0 . 3 1 p p b

at 0.086

of 0.86

and

and 0 . 3 4 p p b

increased

in Fig.

the e x p e r i m e n t a l rapidly

period.

and r e a c h e d

The

a plateau

I.

no clear d e p e n d e n c y but r e m a r k a b l e

indicates

to 2.2-fold.

to carp,

higher

Statistical

the d i f f e r e n c e

butoxide

during

on the e x p o s u r e

differences

between

isomers,

I.

0.43 ppb of the test

increased,

in carp

of d-phenothrin,

fish m e a s u r e d

piperonyl

and 0.084

of i n t o x i c a t i o n

isomer

BCF values

concentrations

When

and 3 . 3 p p b at nominal

of d - p h e n o t h r i n

concentration

All the

and d - t r a n s

respectively.

showed

as shown

of d - c i s

BCF values analysis

to be s i g n i f i c a n t

compound

but no s i g n i f i c a n t

change

than d - t r a n s

(the t test for m a t c h e d at 0.1%

was e x p o s e d

the BCF v a l u e s

for the d - c i s

of the d - c i s

was o b s e r v e d

level.

in the p r e s e n c e isomer

of

clearly

for the d - t r a n s

isomer

2004

104

4.3ppb*

iO3

O,43ppb*

~ "~

~" tO

/

II

"~

d-cis

(hsh)

102

10'

10'

10°

r" o

O

d-trans (water)

~ 10 °

I

0

I

I

14 28

I

~

d-cis (water)

10-2

I

I

42 56

]4

Days

*: N o m i n a l Values

concentrations correspond

3.4 and 0 . 3 4 p p b

Table

I.

I

I

28

I

42 56

Days

B i o c o n c e n t r a t i o n of g e o m e t r i c

Fig.1.

d-trans (water)

10-'

d-cis (water)

I0-'

" ~ d-trans (fish) d-cis (fish)

i s o m e r s of d - p h e n o t h r i n

of d - p h e n o t h r i n

in water.

to 0.86 and 0 . 0 8 6 p p b for the d - c i s

for the d - t r a n s

BCF's of g e o m e t r i c

isomer,

respectively.

isomers of d - p h e n o t h r i n

4.3ppb*

BCF Days

14

28

42

56

cis

0.43ppb*

BCF ratio trans

and

BCF

(cis/trans)

BCF ratio

cis

trans

(cis/trans)

480

330

1.5

490

320

1.5

480

360

1.3

670

500

1.3

450

370

1.2

660

290

2.2

470

320

1.5

730

490

1.5

610

400

1.5

380

280

1.4

530

400

1.3

600

530

1.1

330

270

1.2

500

340

1.5

420

320

1.3

330

210

1.6

*: N o m i n a l

concentrations

Values correspond

of d - p h e n o t h r i n

to 0.86 and 0 . 0 8 6 p p b

0 . 3 4 p p b for the d - t r a n s

isomer,

in water. for the d - c i s

respectively.

and 3.4 and

2005

(BCF c i s / t r a n s piperonyl

ratio:

butoxide

11-20),

was d e t e c t e d

In this e x p e r i m e n t ,

average

isomers

compound

of the test

concentrations observed plateau

of 0.086

actual

2. In e x p o s e d

concentrations

in w a t e r

exposure.

confirmed

in Table

at the c o n c e n t r a t i o n s

and 0.34ppb),

for fish d u r i n g was also

as shown

after

were

of the d-cis

0.080

As shown

and 0 . 3 0 p p b

in Fig.

and d-trans

(at n o m i n a l

of i n t o x i c a t i o n

was

2, a b i o c o n c e n t r a t i o n

exposure.

Table

0.43ppb+PBO"

of 81 to 110ppm.

and no s y m p t o m

2 w e ek s

fish,

2.

Effect

of p i p e r o n y l

butoxide

10"

on the b i o c o n c e n t r a t i o n

~

102

geometric

d-cis (fish) d-trans (fish)

~. o.

isomers

of

of

d-phenothrin

0.43ppb+PBO*

c 10' 0 P

BCF

t(D

o 10° t-

Days

cis

ratio trans

(cis/trans)

O

O

d-trans (water)

~ 10-'

14

d-cis (water)

28 10-2

3800

190

20

3600

320

11

2600

210

12

2400

200

12

Days

*: d - P h e n o t h r i n Fig.2.

Bioconcentration geometric

isomers

d-phenothrin presence

of

to

of

in the

of p i p e r o n y l

d-Phenothrin at 0 . 4 3 p p b

was e x p o s e d

to carp

in

the p r e s e n c e

of 1 . 0 p p m

of p i p e r o n y l

butoxide

(PBO).

in

the

of p i p e r o n y l

butoxide W.

carp

was e x p o s e d presence butoxide

of

at 0 . 4 3 p p b 1.0ppm

(PBO).

2006

DISCUSSIONS

In this isomers

study,

a certain

is o b s e r v e d

statistically difference

difference

(Table

significant

can be done

I). The d i f f e r e n c e at 0.1% level.

by c o n s i d e r i n g

bioconcentrations

of the isomers.

biotransformation

will be d e s c r i b e d

compartment

bioconcentration

BCF : Cf(~)/Cw(~)

where

Cf(~)

water,

and Cw(~)

respectively

rate constants, The e q u a t i o n

in carp was

Elimination

rapidly

isomer

products

isomer

(e.g.

c-phenothrin When

exposure

considerable (Table

reactions

microsomal oxidation

elevation

isomers

in

isomer

as r e p o r t e d

The higher

should

were d e g r a d e d

one week.

lower BCF value

study:

isomer

have

Based on was

similarly

for

of the d - t r a n s

metabolic,

resulted

therefore,

in the

lower

p r o d u c ts

moiety

or h y d r o l y z e d

isomer

conjugate

in carp 3) were ester

of the alcohol

amounting

oxidized

(e.g.

and at the methyl

metabolites

sulfate

conjugate

were of 3-(4'-

to 47% of the i d e n t i f i e d were mostly

of 4'-OH-,

formed

desphenyl-,

from the d - c i s and

4'-OH-~t-alc-

to 77%).

is c o n d u c t e d

in the p r e se n c e

of the BCF value

piperony

butoxide

of piperonyl

is o b s e r v e d

is well

o x y g e n a s e 4) and the d - c i s as m e n t i o n e d

attributable

results

or d - t r a n s - i s o m e r

d-cis

the d - t r a n s

isomer,

Ester-cleaved

acid

glucuronide amounting

Both

of d - p h e n o t h r i n

of the d - t r a n s

whereas

2). Since

by b i o t r a n s f o r m a t i o n .

fish w i t h i n

of 14C,

explains

at 4 ' - p o s i t i o n

hydroxyphenoxy)-benzoic metabolites),

in fish and

isomer.

of the acid moiety.

principal

well

in the p r e s e n t

than the d - c i s

oxidations

from

than the d - c i s

rate of the d - t r a n s

The m a j o r m e t a b o l i c

group

of the two-

and e l i m i n a t i o n

in b i o t r a n s f o r m a t i o n

e t . a l . 3)

excreted

metabolized

cleavage,

rate

of 1 4 C - l a b e l e d

by T a k i m o t o

completely

mammals. 7) This o b s e r v a t i o n

BCF value

by

)

of the c o m p o u n d

K 1 and K 2 uptake

rate c o n s t a n t

the higher

for the d i s t r i b u t i o n

the d - c i s

on

BCF e q u a t i o n

( I

state,

the o b s e r v a t i o n s

than

for the

in fish.

investigated

more

of the c o m p o u n d

= KI/(K2+K3)

that

but

explanation

of m e t a b o l i s m

by a f o l l o w i n g

biotransformation

and almost

disappearance

the effect

and K 3 an e l i m i n a t i o n

The c o m p a r a t i v e

is not so large,

A possible

are the c o n c e n t r a t i o n s

suggests

and d - t r a n s

d-cis

model I)-

at a steady

the lower BCF value

rapidly

in BCF's b e t w e e n

isomer

oxidative

reaction

a

for the d - c i s

known as an inhibitor is m e t a b o l i z e d

above 3) , the e l e v a t i o n

to a reduced

butoxide,

observed

here

isomer of

mainly should

of the i n h i b i t e d

by be

enzyme.

The

2007

contribution calculated eq.(1)

and by s e t t i n g

Thus, d-cis

to the e l i m i n a t i o n

to be 2.3 to 11 by i n t r o d u c i n g

conducted

with

of b i o t r a n s f o r m a t i o n

the rate c o n s t a n t

in the p r e s e n c e

a metabolic

isomer

The d-trans

relations

effect

the o b s e r v e d

K3/K 2) was

BCF values

K 3 to be 0 when e x p o s u r e

into the was

butoxide.

on b i o c o n c e n t r a t i o n

whose metabolism

the d-trans

it f o l l o w e d

of p i p e r o n y l

(i.e.

proceeds

was c l e a r l y

mainly

through

indicated

oxidation

for the

as c o m p a r e d

isomer.

isomer,

on the other

a rapid e s t e r

cleavage

to the o x i d i z i n g

hand,

did

not show such e l e v a t i o n

or hydrolysis,

because

w h i c h has no d i r e c t

enzyme.

CONCLUSION

This

study

clearly

demonstrated

carp was

significantly

presence

of p i p e r o n y l

d-cis

isomer,

higher

that

the BCF value

than the d-trans

butoxide

resulted

but no r e m a r k a b l e

change

of d-cis

isomer

phenothrin

and an e x p o s u r e

in an e l e v a t e d

BCF value

in BCF's was o b s e r v e d

in

in the

for the

for the d-trans

isomer. Considering result

an i n h i b i t e d

obtained

oxidative

here was well

bioconcentration

phenomenon

metabolism

explained

for the d-cis

and a s i g n i f i c a n c e

isomer,

the

of m e t a b o l i s m

in

was e x e m p l i f i e d .

ACKNOWLEDGMENTS

The a u t h o r s and Dr.

are i n d e b t e d

H. Yamada,

to P r o f e s s o r

Sumitomo

Chemical

Dr. T. Nishihara,

Co.,

Ltd.,

Osaka

University

for their helpful

discussions.

REFERENCE

I) Lech,

J.J.

2) Melancon,

and J.R. M.J.,Jr.

Bend,

Environ.

and J.J.

Lech,

Health

Perspect.,

Xenobiotica,

34,

7(10),

115-131

633-640

3) Takimoto, Y.,et.al., A b s t r a c t paper of the S i x t h I n t e r n a t i o n a l of P e s t i c i d e C h e m i s t r y (IUPAC) Otawa, 7B-04 (1986). 4) Casida, 5) OECD

J.E.,et.al.,

(1981),

6) Bligh, E.G. (1959). 7) Izumi

Science,N.

Guidelines and W.J.

,et.al.,

Y.,

for T e s t i n g

Dyer,

J. Pestic.

Can. Sci.,

153,

1130-1133

9, 259-267

(Receivedin Germany 18 May 1992; accepted25 May 1992)

and Physiol., (1984).

Congress

(1966).

of C h e m i c a l s . , v o l . 3 0 5 C ,

J. Biochem.

(1980).

(1977).

Paris. 37,

911-917