Base-neutral extractable organic pollutants in biota and sediments from Lake Pontchartrain

Chemosphere, Vol.14, No. lO, pn 1561-1569, Printed in Great Britain

Base-Neutral

1985

0045-6535/85 $3.00 + .OO 01985 Peraamon Press Ltd.

Extractable Organic Pollutants in Biota and Sediments from Lake Pontchartrain

Jo Ann McFall,

Shelley R. Antoine,

and Ildefonso R. DeLeon

Center for Bio-Organic Studies, University of New Orleans, Lakefront, New Orleans, Louisiana 70148 ABSTRACT. A study was conducted to screen the three passes that link Lake Pontchartrain to the Gulf of Mexico via Lake Borne for the presence of EPA base-neutral (BN) priority pollutants and any other pollutants detected in significant concentration. Biota and sediment samples were collected and analytical procedures were developed for the trace analysis of BN organics in these matrices. Compounds identified include alkanes (normal, branched, cyclic), alkenes, aromatics, alkylated aromatics, polynuclear aromatic hydrocarbons and their alkylated derivatives, phthalates, ketones, furans, thiophenes, phenols, amines, nitriles, thiazoles, amides, aldehydes, alcohols, free fatty acids, fatty acid methyl and ethyl esters, phosphates, sterols. Concentrations were in the parts-per-billion range. INTRODUCTION For several decades the inland and coastal

aquatic ecosystems

recipients of a multitude of man-made and naturally-occurring stances.

This is a consequence of increasing population

development of industrial and agricultural

activity.

chemical

sub-

and the accompanying

Many of these materials,

their by-products and degradation products are ultimately aquatic environment.

have been the

The degree to which these pollutants

released

into the

impact the environ-

ment and its inhabitants depends largely upon their quantity and nature. As part of continuing pollutants

investigations

in natural bodies of water

Mc Fall et al. 1979,

to characterize

(Laska et al.

1976,

and identify chemical Laseter et al. 1978,

Laseter and Ledet 1979, Overton and Laseter 1980, Overton

et al. 1980, Laseter et al. 1981, and Ferrario et al.

1985) and as part of a

preliminary study of the nutrient and toxic substances passes into Lake Pontchartrain,

chemistry

for the three

we report here the results of biota and sediment

samples screened for base-neutral priority pollutants. Lake Pontchartrain 1631km 2 estuary located passes, Canal

The Rigolets,

(New Orleans,

is a shallow,

in the deltaic plain of the Mississippi

oligohaline, River.

Three

the Chef Menteur Pass, and the Inner Harbor Navigation

(IHNC) provide an indirect,

The Rigolets accounts

Louisiana)

restricted

connection

for 44 percent of water transport

1561

to the Gulf of Mexico. into and out of the

1562 lake;

the Chef Menteur Pass and the IHNC, for 32 and 6 percent,

respectively.

Other

inputs

run-off

overflow

include rivers,

bayous, municipal and agricultural

from the M i s s i s s i p p i

metropolitan

area of over

River during times of potential

Stone

A

1.5 million people is adjacent to the lake and the

lake serves as a m a j o r recreational q u a n t i t i e s of crabs,

and the

flooding.

area and as a source of s u b s t a n t i a l

shrimp and other aquatic foods

(McFall et al.

1979 and

1980).

M A T E R I A L S AND METHODS Samples of oysters

(Crassotrea vir~inica),

clams

(Ran@ia cuneata),

s e d i m e n t s were collected

in May-June

N a v i g a t i o n Canal

the Chef Menteur Pass, and The Rigolets,

Pontchartrain.

(IHNC),

and

1980, from the mouths of the Inner Harbor at Lake

All samples were packed in ice, frozen and kept at -5°C until

analysis. Extraction Methods Five g of tissue (pH 12-13) with

(oysters or clams) were a c c u r a t e l y weighed

6N NaOH.

and m a d e basic

Five hundred ml of diethyl ether was then added and

the sample was spiked with ten ug each of the recovery standards, anthracene

and N - n i t r o s o - N - p h e n y l - b e n z e n a m i n e .

disruptive

extraction

homogenizer;

in diethyl

the h o m o g e n i z e r

and the w a s h i n g s combined

The samples were s u b j e c t e d

ether with a Brinkman Polytron

blades were subsequently rinsed with d i e t h y l

(200-400 mesh S-X12 Bio-Beads ®) connected

into a boil-

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

e x t r a c t was fractionated by gel permeation column chromatography. columns

ether

The h o m o g e n i z e d

tissue s u s p e n s i o n was c e n t r i f u g e d and the supernatant was d e c a n t e d to I-2 ml on a rotary evaporator.

to

tissue

with the original tissue homogenate.

ing flask and c o n c e n t r a t e d

2-methyl-

Two

in series, p r o v i d e d

50 x I cm acceptable

s e p a r a t i o n of the m a j o r i t y of the fatty acid methyl esters from the b a s e - n e u t r a l components monitor (TBN)

(ISCO model U-A5 or model

the columns e f f l u e n t s during

T840 absorbance monitors were used fractionation).

The total b a s e - n e u t r a l

fraction was eluted with dichloromethane and concentrated

rotary evaporator. ferred

of 200 ul.

concentrator

Each total b a s e - n e u t r a l

n a n o g r a m s each of the internal prior

to 2-3 ml on a

This 2-3 ml total base-neutral organic fraction was

to a m i c r o - e v a p o r a t i v e

to

for c o n c e n t r a t i o n

trans-

to a final volume

fraction concentrate was spiked w i t h

standards,

10,000

cis-decalin and d 1 0 - a n t h r a c e n e ,

to final analysis by combined gas c h r o m a t o g r a p h y - m a s s

spectrometry

(GC/MS). Sediment mouth,

m a d e basic ether.

samples

(approximately

100 g wet weight)

were placed

p i n t - s i z e Mason jars equipped with a Teflon lined cap.

in wide-

The s a m p l e was

(pH 12-13) with 6N NaOH prior to the addition of 200 ml of diethyl

Each sample was spiked with ten ug each of the recovery standards,

2-methylanthracene extracted

and N - n i t r o s o - N - p h e n y l b e n z e n a m i n e .

for 3 hrs before

s u p e r n a t a n t was d e c a n t e d a l i q u o t of

The sample was

centrifugation at 500 rpm for 10 min.

into a boiling flask and reserved

100 ml of diethyl

ether was added,

The

for future

and the sediment

tumbleuse.

An

tumble-extracted

1563

again for 3 hrs.

Following

into the b o i l i n g

flask,

centrifugation

a third wash

and d e c a n t a t i n g

(100 ml) of d i e t b y l

sample was m a n u a l l y

agitated

and the e t h e r

to the first and second extracts.

added

e x t r a c t was r e d u c e d anhydrous

sodium

to a p p r o x i m a t e l y

sulfate.

The total b a s e - n e u t r a l rotary evaporator

column

still

n a n o g r a m s each of the internal

to a m i c r o - e v a p o r a t i v e

Gas C h r o m a t o g r a p h y

Hewlett-Packard

operated

to 2-3 ml

concentrator

To d e t e r m i n e

and Mass S p e c t r o m e t r y

in a forced-air

(GC/MS)

7920 multi-disc drive

7970B tape deck. coated with SE-52

temperature-programmed Sample

for con10,000

the dry weight of the dried

5985A GC/MS system

data storage system and a

The column was a 50 m x 0.3 mm ID glass (Supelco Inc.).

The

injection port was

in the s p l i t l e s s mode at 250°C with a 30 sec vent delay.

of 32 min.

in a

and reweighed.

a Hewlett-Packard

capillary column

was

standards, d10- a n t h r a c e n e and cis-decalin,

C o m b i n e d G C / M S was done on a Hewlett-Packard Model e q u i p p e d with

in diethyl ether,

Each sample was spiked with

an aliquot of each sample was weighed,

c h e m i c a l v a p o r hood,

and dried with

(30 x 2 cm) and eluted with d i c h l o r o m e t h a n e .

to a final volume of 200 ul.

samples,

it was centrifuged

fraction was collected and c o n c e n t r a t e d

and t r a n s f e r r e d

The

The total base-neutral

2 ml on a rotary evaporator

prior to final a n a l y s i s by combined GC/MS. sediment

5 min before

The concentrated extract,

loaded o n t o a gel p e r m e a t i o n

centration

for approximately

of the second extract

ether was added.

The oven was

from 50°C to 250"C at 4°C/min with a minimum

final hold

ionization was by electron impact at 70 eV with a source

temperature

of 200°C.

1.52 sec).

The G C / M S data g e n e r a t e d on the H e w l e t t - P a c k a r d

The mass range was scanned from 45 to 450 amu (cycle time

treated on a F i n n i g a n / I N C O S

2000 data system by means

for EPA p r i o r i t y p o l l u t a n t b a s e - n e u t r a l s

5985A GC/MS were

of user prepared

and a general

routines

search for other

organics.

RESULTS AND D I S C U S S I O N The r e s u l t s of the a n a l y s i s of oyster samples from the IHNC and of clam samples

from the Chef Menteur Pass and The Rigolets are presented

and 2.

The o y s t e r s

base-neutral

from the IHNC population contained

p r i o r i t y pollutants.

in Tables

The majority of the base-neutral organic

priority pollutants

detected

pooled clam s a m p l e s

from the Chef Menteur Pass and The Rigolets contained

and ten b a s e - n e u t r a l samples,

The g e n e r a l

were polynuclear a r o m a t i c

p r i o r i t y pollutants,

the p r e d o m i n a n t

respectively.

from the three p a s s e s

revealed

which were

at least to chemical class.

following cyclic), thiazole,

aromatics,

aldehydes,

acid ethyl e s t e r s

(PAHs).

The

nine

As with the IHNC oyster

PAHs, phenols,

alcohols,

and sterols.

for oyster and clam samples

139 distinct n o n - p r i o r i t y pollutant compounds

c l a s s e s of compounds were detected: alkenes,

hydrocarbons

b a s e - n e u t r a l organic p o l l u t a n t s were PAHs.

search of the GC/MS data developed

identified

I

twenty specific EPA

Representatives

alkanes

amines,

free fatty acids,

(normal,

ketones,

of the

branched,

nitriles,

and

a

fatty acid methyl esters,

fatty

1564

Table

I.

Base-neutral organics detected Pontchartrain at passes

in biota samples

from Lake

nano@ram/~ram (ppb) wet weight Chef IHNC Menteur Rigolets O~ster a Clams b Clams b

Compound Priority Pollutants c bis(2-chloroethyl)ether

0.6

dichlorobenzene bis(2-chloroisopropyl)

_d

58. ether

isophorone

0.8

130.

-

38. 12.

bis(2-chloroethoxy)methane

140.

1,2,4-trichlorobenzene naphthalene

35.

120.

51.

2-chloronaphthalene

34.

970.

140.

acenaphthylene

36.

34.

130.

dimethyl phthalate

8.4

acenaphthene

46.

fluorene

44.

-

24.

46.

450.

340.

21.

diethyl phthalate

1100.

phenanthrene

220.

anthracene

44.

di-n-butyl phthalate

72.

-

36.

43.

240.

200.

630.

330.

570.

fluoranthene

80.

pyrene

200.

benz(a)anthracene

9.9

chrysene

58.

bis(2-ethylhexyl)phthalate benzo(k)fluoranthene

210. 12.

Non-Priority Pollutants e methylnaphthalene

15.

2-chlorobenzamine

41.

methylphenanthrene 1,2-benzenedicarboxylic C 4 alkylated

178

2.8 acid

-

aMean of eight samples. bResults are of a composite sample. CQuantitated by authentic standards; adjusted for percent recovery. d,,_- means not detected. eConcentrations expressed as d10-anthracene equivalents; adjusted for percent recovery.

57

1565

Table 2.

Chemical

Other o r g a n i c s detected in the base-neutral from Lake P o n t c h a r t r a i n at passes

fraction of biota

samples

c o n c e n t r a t i o n ranges in n a n o g r a m / g r a m (p~b) wet weight a Chef Number of IHNC Menteur Rigolets components O[ster b Clams c Clams c

classes

alcohols

4

990-5400

aldehydes

4

100-1800

21-250

45-410

160

26

alkanes

30

32-2600

alkenes

4

300-7200

alkylbenzenes

3

-

11-31

amines

I

-

ii0

28

b r a n c h e d alkanes

I

470

34

320

cyclic alkanes

5

fatty acids fatty acid ethyl e s t e r s fatty acid methyl esters

11-720

16-720

_d

-

34-1200

18-94

-

11

79-64000

2

510-19000

52

19-42000

-

190-39000

35-19000

1400

1200

8.0-8700

7.6-5200

42-490

23-220

ketones

4

640

nitriles

I

-

phenols

3

33

27-130

16-66

sterols

8

130-25000

70-2500

63-2600

thiazoles

I

ii

-

89

a C o n c e n t r a t i o n s e x p r e s s e d as d 1 0 - a n t h r a c e n e equivalents; percent recovery. bMean of eight samples. C R e s u l t s are of a c o m p o s i t e sample. d-_. means not detected.

The results of the analysis of the sediment samples are given

in Tables

3 and 4.

p o l l u t a n t s were identified the largest number replicate

IHNC o y s t e r

Rigolets contained

adjusted

for

from the three passes

specific b a s e - n e u t r a l o r g a n i c p r i o r i t y

in these analyses;

(twenty-three)

detected were PAHs.

respectively.

Twenty-five

23

the IHNC sediment samples showed

of the priority pollutants.

As was true

in the

samples, most of the base-neutral priority p o l l u t a n t s The sediment samples from the Chef Menteur Pass and The

eleven and eight base-neutral priority pollutants,

Most of the base-neutral organic priority p o l l u t a n t s

found were

PAHs. The general

search of the GC/MS data

for the sediment

three passes revealed

a total of 178 individual

which were identified

at least to chemical

following

class.

classes of compounds were detected:

cyclic),

alkenes,

ketones,

furans,

alcohols, phenols,

acid methyl esters,

aromatics,

thiophenes,

samples

alkanes

thiazoles,

compounds

R e p r e s e n t a t i v e s of the (normal,

branched

PAHs, alkylated aromatics,

a fatty acid ethyl ester,

from the

non-priority p o l l u t a n t

amides,

and

phthalates,

free fatty acids,

and a phosphate.

fatty

1566

Base-neutral organics detected Pontchartrain at passes

Table 3.

in sediment

samples

from Lake ,

m

nanogram/gram (ppb) dry weight Chef Menteur b Rigoletsb IHNC a

Compound Priority p011utantsC isophorone

0.9

1,2,4-trichlorobenzene

_d

naphthalene

12.

10. 3.4

-

0.9

3.4

3.0

2-chloronaphthalene

1.6

3.0

7.6

acenaphthylene

0.4

1.7

6.5

dimethyl

0.2

2.0

phthalate

acenaphthene

17.

fluorene

1.6

5.3

diethyl phthalate

25.

hexachlorobenzene

-

phenanthrene

51.

anthracene

14.

di-n-butyl

65.

phthalate

4.5

15.

fluoranthene

42.

25.

pyrene

41.

63.

benzo(a)fluorene

21.

benzo(ghi)fluorene benz(a)anthracene

38. 54.

butyl benzyl phthalate phthalate isomer

a benzo fluoranthene

isomer

7.3

0.8 56.

a benzo fluoranthene

-

210.

86. 2.9

benzo(e)pyrene

0.6

benzo(a)pyrene

0.5

Non-priority

6.0

6.5

chrysene di-n-octyl

7.9

pollutants e

5-methyl-l-phenyl-hexanone C10-alkane ethyl benzoate

39. 0.5 65.

5-(1-propenyl)-1,2benzodioxole

8.7

2-methylnaphthalene

8.7

1-methylnaphthalene dimethyl

tetralin

dimethylnaphthalene

6.0 2.8 2.9

1-butylhexylbenzene dibenzofuran diethyl

phosphate

42. 1.9 3.6

1567

Table 3, continued nano@ram/@ram (ppb) dry weight Chef IHNC a Menteur b Ri~91etsb

Compound 5,6,7,7A-tetrahydro-4,4,7Atrimethyl-2(4H)benzofuranone trimethyl

6.3

naphthalene

4.2

0.1 -

180.

1-pentylheptylbenzene

-

110.

1-butyloctylbenzene

-

76.

1-propylnonylbenzene

-

55.

1,1-diethylpropylbenzene 1,2-dimethyl-4-phenylmethylbenzene

1.8

nonyl phenol

7.9

6,10,14-trimethyl-2pentadecanone

2.8

methyl phenanthrene/ methylanthacene

15.

phenylnaphthalene 4H-furo 3,2-G/I/ benzopyran-4,7,9-trione

10. 11.

dimethyl phenanthrene/ dimethyl anthracene

7.1

methyl pyrene/methyl fluoranthene

3.8

octadecenoic

acid ethyl ester

40.

benzo(b)naphthothiophene

1.8

methylbenz(a)anthracene

1.6

N-ethylpentamide

0.7

C29-alkane

2.0

14.

aMean of eight samples. bResults are of one sample. CQuantitated by authentic standards; adjusted for percent d,,_- means not detected. eConcentrations expressed as d10-anthracene equivalents; adjusted for percent recovery.

While the EPA base-neutral sediment

samples

compounds decalins furans,

are of obvious

identified

are probably

and tetralins, benzofurans

recreational

priority

cyclic

pollutants

anthropogenic biogenic.

and branched

and thiophenes

craft and commercial

utilize

in the biota and

the majority of the other

The presence alkanes,

are indicative vessels

found

origin,

recovery.

of alkylated

PAH's,

of fossil

benzenes,

alkylated PAH's, fuels.

these waterways,

Since small these find-

ings are not u n u s u a l . The base-neutral EPA priority pollutants identified in the biota and sediment have quite variable physical, chemical, physiological, and toxicological properties

which allow a variety of commercial

and industrial

applications.

1568

These chemicals are used extensively as chemical solvents,

lubricants, degreasing

pharmaceuticals,

etc.

raw materials,

agents, plasticizers,

identified

sediment samples have possible adverse human health effects nervous system depression,

teratogenic effects,

mutagenic

various renal and hepatotoxic effects which may manifest 1984).

(Christensen,

1975; Clayton and Clayton,

At the present time maximum permissible

organisms are not available.

tissue

However, benzo(a)pyrene

suspected carcinogens and therefore no safe exposure established based on the non-threshold

assumption

potentially have an effect on the biological these compounds tend to accumulate solubility.

including central

effects,

narcosis and

at very low levels of

1981, Moore and Karosek, concentrations

for aquatic

and hexachlorobenzene

are

limit can rationally be

processes

in biological

in the biota and

(EPA 1980). These organics in the environment since

systems due to their high lipid

These organics are not only toxic to the aquatic organisms present

in Lake Pontchartrain but can also affect the health deriving

fungicides,

(Clayton and Clayton 1981).

All the base-neutral EPA priority pollutants

the compounds

intermediates,

insecticides,

and well-being of humans

food products from this lake due to the accumulation

tion of these organics

and biomagnifica-

in the higher trophic levels.

The data which have been generated

in this study show the presence of a

significant number of base-neutral organic priority pollutants neutral organics of industrial

and biogenic origin

sediment samples from Lake Pontchartrain. affect the ecology of Lake Pontchartrain

and other base-

in benthic organism and

The degree

to which these pollutants

is not known and cannot be properly

assessed until further investigations are conducted. ACKNOWLEDGEMENTS The authors gratefully Army Corps of Engineers,

Holmes, J.L. Laseter, G.C. assistance,

acknowledge partial

Contract

support of this work by the U.S.

#DACW29-82-M-0189.

Lawler, J.M. Pachankis,

and K.D. Trembley for assistance

We thank C.J. Byrne, J.P.

and A.H.

Rhyans for technical

in the preparation of this manu-

script. REFERENCES Christensen HR, Luginbyhl TT, Benigna CS (eds.) (1975) Registry of toxic effects of chemical substances. U.S. Department of Health, Education and Welfare Clayton GD, Clayton FE (eds.) (1981) Party's industrial Volume 2B. John Wiley and Sons Inc., New York EPA (1980) Ambient water quality criteria, 049, 056, 059, 066, 067, 069)

hygiene and toxicology,

EPA 440/5-80(015,

028, 030, 031,

Ferrario JB, Lawler GC, DeLeon IR, Laseter JL (1985) Volatile organic pollutants in biota and sediments of Lake Pontchartrain. Bull Environ Contamin Toxicol 34:246-255 Laseter JL, Ledet EJ (]979) Hydrocarbons and free fatty acids associated with the air/water interface, sediments, and beaches of the Timbalier Bay and Offshore Louisiana Area. Rice Univesity Studies 65:265-286

1569

Laseter JL, Lawler GC, Overton EB, Patel JR, Holmes JP, Shields MI, Maberry MA (198]) Characterization of aliphatic and aromatic hydrocarbons in flat and Japanese type oyster and adjacent sediment collected from L'Aber Wrac'h following the AMOCO CADIZ Oil spill. In AMOCO CADIZ fates and effects of the oil spill. CNEXO, Paris, 633-644 pp Laseter JL, DeLeon IR, Remele PC (]978) Determination of mirex in fish by gas chromatography and gas chromatography-mass spectrometry-computer system. Anal Chem 5 0 : 1 1 6 9 - 1 1 7 2 Laska AL, Bartell CK, Laseter JL (]976) Distribution of hexachlorobenzene and hexachlorobutadiene in water, soil, and selected aquatic organisms along the lower Mississippi River, Louisiana. Bull Environ Contam Toxicol 15:535-542 McFall JA, Huang WY, Laseter JL (]979) Organics at the airwater Lake Pontchartrain. Bull Environ Contam Toxicol 22:80-87

interface of

Moore RA, Karasek FW (1984) GC/MS identification of organic pollutants Caroni River, Trinidad. Interm J Environ Anal Chem 17:203-221

in the

Overton EB, Laseter JL (1980) Distribution of aromatic hydrocarbons in sediments from selected Atlantic, Gulf of Mexico, and Pacific Outer Continental Shelf Areas. Petroleum in the Marine Environment ]85:327-341 Overton EB, Mascarella SW, McFall JA, Laseter JL (1980) Organics in the water column and air-water interface samples of Mississippi River water. Chemosphere 9:629-633 Stone JH (ed) (]980) Environmental analysis of Lake Pontchartrain, Louisiana, its surrounding wetlands, and selected land uses. Center for Wetland Resources, LSU, Baton Rouge, LA, I:]-60

(Received in Germany 28 July 1985; accepted 21 August 1985)