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DEVELOPMENT OF BIOASSAY PROCEDURES FOR
DEFINING POLLUTION OF HARBOR SEDIMENTS
PART 1.
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EPA 60.0/3-81-025
March 1981
FINAL REPORT
DEVELOPMENT OF BIOASSAY PROCEDURES FOR
DEFINING POLLUTION OF HARBOR SEDIMENTS
PART I.
by
DONALD A, BAHNICK
WILLIAM A. SWENSON
THOMAS P. MARKEE
DANIEL J. CALL
CRAIG A. ANDERSON
R. TED MORRIS
Center for Lake Superior Environmental Studies
University of Wisconsin, Superior, Wisconsin
PROJECT NUMBER: R804918-01
PROJECT OFFICER: RICHARD ANDERSON
ENVIRONMENTAL RESEARCH LABORATORY
DULUTH, MINNESOTA
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CONTENTS
Section 1 - Introduction . ....... ii
Section 2 - Conclusions iii
Section 3 - Recommendations. v
Section 4 - Overview of the Study 1
Background and Purpose 1
Biological Tests to Screen Sediments. . . . . 3
Daphnia Bioassays. ..... . 3
Hexagenia Bioassays. ...... 4
Pontoperia Bioassays 5
Fish Behavior Bioassay 5
Sediment Employed and Chemical Characterization 6
Uptake of Chemicals by Chironomids and Hexagenia. ........ 7
Bioaccumulation Determinations Using High Liquid Pressure
Chromatography 7
Chemical Correlations to Bioassays Using Biological Probes. ... 8
Development of Harbor Sediment Sampler 8
Section 5 - Study Area . 10
General Harbor Description ; 10
History of Pollution 12
Physical Nature of the Sediments 13
Chemical Nature of the Sediments 14
Aquatic Life. 16
Section 6 - Project Field Sampling 18
Sampling Sites 18
Sediment, Water and Chironomids Sampling 19
Hexagenia Limbata . 21
Pontoporeia Affinis 21
Lepomis Macrochirus 23
Section 7 - Chemical Characterization of Samples and Water -
Sediment System Preparation . 24
Sediment-Water Systems Utilized ..... 24
Treatment of Core Samples 24
Background . 24
Procedure 25
Sediment Analysis 27
General Physical and Chemical Sediment Tests 28
Preparation of Sediment Extracts for Metal Analysis 28
Exchangeable Phase Metals 32
Easily Reducible Phase Metals , . 32
Organic and Sulfide Phase Metals 33
Moderately Reducible Phase Metals . 33
Residual Phase Metals ........ 34
Metal Analysis Procedures 35
Metal Analysis Results 35
Analysis for Total Metals in Sediments .... 41
Pesticide and PCB Analysis 41
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Sediment Extraction Procedure. ............... 44
Extract Clean-Up Procedure ................. 44
Separation of PCBs and Pesticides. . 45
Gas Chromatographic Analysis 45
Chlorinated Phenol Analysis ...... 46
Analysis Procedure . 46
PAH Analysis 48
Analysis Procedure . . .......... 49
Analytical Results ..................... 50
Discussion of Sediment Results. ................ 53
Metals in the Sediments. .................. 54
Trace Organics in the Sediments 55
Overlying Water 56
Interstitial Water ..... 59
Elutriate Water 63
Preparation of Liquid Phase (Elutriate Water) and Particulate
Phase 64
Liquid Phase (Elutriate Water) and Particulate Phase Tests. . . 65
Survey of Chemical Properties of Elutriate Water 65
Generated Pore Water 70
Generated Pore Water Production Procedure 71
Generated Pore Water Results. ..... ..... 72
Discussions of Results. ..... 74
Section 8 - Bioassay Tests. . . ............. 83
General Bioassay Procedures 83
Light and Temperature Control ........ 83
Daphnia Culturing Techniques 83
Acclimation and Handling of Test Organisms 84
Oversediment Bioassay. .... 86
Methods 86
Hexagenia-Daphnia Bioassay Methods 86
Pontoporeia affinis Bioassay Methods 90
Results 91
Hexagenia Bioassays 91
Daphnia Bioassays. . 94
Pontoporeia Bioassays. . 98
Elutriate, Interstitial and Pore Water Bioassays 100
Methods 100
Daphnia Bioassay 100
Fish Bioassays 101
Results 106
Daphnia Bioassays 106
Fish Bioassays . 109
Comparisons of Results to Other Sediment Bioassay Studies 110
Section 9 - Bioaccumulation Potential of Sediment Associated Chemicals 114
Chemical Analysis of Hexagenia and Chironomids 114
General Methods 114
Results 115
Bioaccumulation of PCBs 116
Bioaccumulation of Pesticides 116
Bioaccumulation of PAH Compounds 118
Bioaccumulation of Metals. 121
Related Studies 122
Sampling Screening for Organic Chemicals Using HPLC 123
Methods and Samples 124
Results 125
Discussion 128
> -b
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Section 10 - Relationships Between Chemical Characteristics of the
Sediment Systems and Bioassay Toxicity .... 130
Site Location and Toxicity .................... 130
General Site Chemistry and Toxicity. ......... . 131
Specific Chemicals and Toxicity. ................. 133
References. ... . ............... 139
Appendices. . . . 145
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SECTION 1
INTRODUCTION
This study was undertaken to evaluate bioassay techniques which might be
applicable to assessing potential harmful effects resulting from the dredging
and disposal of harbor sediments. Sediments from the Duluth, Minnesota and
Superior, Wisconsin harbor area were used in preparing systems containing water
overlying a sediment substrate, containing interstitial water, or containing
elutriate water. Acute 96 hour toxicity tests were carried out by exposing
Hexagenia 1imbata, Daphnia magna and Pontoporeia affinus to certain of these
systems. Bluegill sunfish (Lepomis macrochirus) were monitored for cough fre-.
quencies and breathing patterns in sediment interstitial water mixed with Lake
Superior water.
Sediment quality was chemically evaluated by extensive analysis of the
sediments and their interstitial waters for a variety of chemical parameters.
The chemical testing included determinations of metals, certain inorganic non-
metallic substances, particle size, pH, Eh and trace organics (PCBs, pesticides,
and PAH compounds). The results of the chemical analyses were compared to the
biological toxicity tests when possible.
Chironomids collected from various harbor sites and Hexagenia 1imbata ex-
posed for 96 hours to harbor sediments were analyzed to determine accumulation
of specific organic compounds (PCBs, PAH and pesticides) and some heavy metals.
Harbor sediments, harbor chironomids and the sediment exposed Hexagenia
1imbata were extracted into organic solvents. The lower molecular weight
organic components in the extracts were separated by reverse phase high pres-
sure liquid chromatography. This chromatographic study was investigated for
its possible use in evaluating the presence of organic compounds having high
lipid bioaccumulation potential.
Part II of this project evaluated a method of assessing sediment quality
by exposing specific enzymes to extracts from the sediments and monitoring
changes in enzyme activity.
ii
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SECTION 2
CONCLUSIONS
On the basis of one or more chemical parameters, most of the sediment
samples used in the bioassay tests would be classified as polluted according
to the currently used sediment evaluation criteria given in Appendix D.
Ranking the sediments according to their concentrations of a large number
of metals, inorganic non-metals and organic chemical parameters indicated
that sites located in the Superior harbor (near the Superior entry to Lake
Superior) and a Lake Superior site were less polluted than sites located near
the more industrialized zones in the harbor.
Chemical analysis for heavy metals in the sediments revealed that the
residual phase of the sediments contained the highest concentrations of most
of them. However measurable amounts (>1 mg/kg) of arsenic, cobalt, copper,
nickel and zinc were found in the organic and sulfide sediment phases for nearly
all the samples and selenium and cadmium were found in these phases for some
of the samples.
PCB concentrations in the sediments ranged from 0.3 to 2.1 mg/kg based
on a dry sediment basis. These values are not high compared to sediments
from other harbor areas. In addition to PCBs, low levels of polycyclic aromatic
hydrocarbons were found in two of the harbor sediments and low levels (1 to
15 yg/kg) of pentachlorophenol were detected.
Studies on determining the amounts of chemical species which would be
released upon flushing the sediments with Lake Superior water showed that only
about one per cent or less of most of the chemicals (COD, Fe, Mn, Ni, Pb, Cu,
Zn, Hg) were removed from the sediments by water extraction. These results
indicated that the measured chemical species were not readily available to
water except when associated with particulates.
Chemical analysis of sediment interstitial water showed that many of the
chemical species were probably associated with very fine (possibly colloidal)
particles in the water. The concentrations of many of the metals were much
lower in filtered water samples compared to non-filtered samples.
The concentrations of chemicals found in liquid phase elutriate water
did not change greatly when prepared from an exposed sediment or from sediment
unexposed to air.
Acute toxicity tests using sediment-water systems resulted in generally
low toxicity to Hexagenia limbata, Daphnia magna and Pontoporeia affinis.
The high survival of tests animals indicated low levels of available toxicants
in the sediments. In 96 hour bioassays, survival was found to be significantly
lower for test sites compared to controls in only a few tests. The results
demonstrated that the animals could be successfully maintained in the complex
test systems and the sediment-water systems caused low acute toxicity. This
low toxicity in combination with low precision between replicates generally
resulted in finding no significant differences in animal survival between
test and control. Among the test species employed, Daphnia magna appeared
to be the most sensitive to toxicants.
iii
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The average cough frequencies of bluegills in dechlorinated city water
and in interstitial water from the sampling site sediments mixed with Lake
Superior water were generally similar. Cough frequencies during the first
22 to 26 hours of fish exposure were found to be elevated above the frequencies
observed for the control for three of the six sites studied. Bluegill opercular
activity was nearly continuous in dechlorinated city water in contrast to broken
patterns of activity observed in interstitial water - Lake Superior water mix-
tures formed from sediments obtained from five of the six sampling sites.
Although survival of test organisms was generally high during the 96
hour toxicity tests, some correlations of survival of Daphnia magna in water
overlying sediments (1977 tests) with chemical parameters were found. Many
of these correlations involved the concentrations of metals in the sediment,
in interstitial water removed from the sediment or in elutriate water formed
by mixing sediment with Lake Superior water.
Some correlations were found between Daphnia magna survival in elutriate
water - Lake Superior water mixtures and individual chemical parameters. Cor-
relations were found between iron concentrations in interstitial water re-
moved from the sediments and Daphnia survival.
A general index of toxic effects, developed by considering the relative
percentage of low survival for the various acute toxicity tests, showed that
survival was generally lower in test systems derived from sediments in the
industrialized areas of the harbor compared to the less developed areas and
the Lake Superior site. Comparing the percent low survival values for the
sites to rankings based on sediment and interstitial water chemical results
indicated a positive correlation to the rankings based on sediment chemistry
(r = 0.80, P > 0.1). A lower correlation coefficient was found between rank-
ings based on interstitial water chemistry and the percent low survival values.
These results indicate that the combined chemical tests for the sediments were
a fair indicator of general toxicity.
Evidence was found that chironomids dwelling in harbor sediments had ac-
cumulated PCBs and p,p'-DDE. Similar results were found for Hexagenia limbata
exposed to harbor sediments for 96 hours.
Chromatograms of organic extracts from sediments, chironomids and Hexagenia
1imbata, using reverse phase high pressure liquid chromatography, showed the
presence of organic compounds with bioaccumulation potential. This method
of screening samples for the amounts of bioaccumulated organic compounds is
potentially useful.
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SECTION 3
RECOMMENDATIONS
The use of Daphnia magna, Hexagenia llmbata or Pontoporeia affinis as
test organisms for potential toxic effects of sediments should be considered
further. Of these species, Daphnia magna appears to be most suited as a test
organism due to ease of culturing, sensitivity and the large amount of in-
formation available on the response of Daphnia to specific chemicals. Further
tests would be useful employing sediment samples having greater variation in
chemical quality. Recognizing that sediments used in this study contained
large quantities of potentially toxic heavy metals in unavailable or nontoxic
forms, it is important to develop better understanding of the conditions which
would result in transformation to available forms and the effects that such
transformations would have. Comparisons of toxicity results to other recently
developed toxicant screening techniques such as algal or luminescent bacteria
assays are recommended.
It was necessary to design the tests following approved criteria for
ecological evaluation of dredging and dredge spoil disposal in marine systems.
Following these criteria, controls for the bioassay tests were derived from
sediments from.Lake Superior and a relatively undisturbed area of the harbor.
Because these sediments contained varying quantities of many toxic substances,
their use negated accurate determination of the sensitivity of the bioassay
procedures. It is therefore recommended that future studies aimed at identify-
ing screening procedures incorporate more effective controls.
Based on our observations, bluegill cough frequencies are difficult to
interpret and their usefulness in determining differences in sediment quality
was limited due to observed similarities in results for the various sites.
The data suggests that extensive experience in conducting fish cough response
tests is necessary to interpret the results, and therefore the technique has
limited applications as a general screening procedure.
Further analysis of bioassay results and chemical characteristics of the
sediments is desirable. If correlations hold for a wide variety of sediments,
additional understanding of the causes of the observed toxicity results may
be obtained.
Screening of sediment extracts or extracts of animals exposed to sediments
using reverse phase high pressure liquid chromatography should be further
investigated. In particular, the eluent fractions containing organic compounds
with high lipid solubility should be studied for methods to quantitate and
perhaps identify highly bioaccumuable compounds.
v
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SECTION 4
OVERVIEW OF THE STUDY
BACKGROUND AND PURPOSE
The need for maintaining vessel accessibility to our nation's waterways
necessitates a continuous dredging program by the Corps of Engineers, The
enormity of this program is demonstrated by the 290 million cubic meters of
sediment dredged annually in the United States, About 65% of this dredged
material is deposited in streams, lakes and coastal waters (Boyd e_t al_., 1972).
The prediction of ecological perturbations due to dredging activities is
difficult. The need for a reliable evaluation procedure to identify the
potential effects of chemical contaminants in dredged or fill materials on
water quality and the aquatic community is recognized.
Section 103 of Public Law 92-532 (Marine Protection, Research and
Sanctuaries Act of 1972) requires the evaluation of dredged material pro-
posed to be discharged into ocean waters. Guidelines for this evaluation
have been presented in the Federal Register, Vol. 42, Mo. 7, Tuesday, 11
January 1977. An implementation manual for this evaluation has been pre-
pared (EPA/Corps of Engineers, 1977) which incorporates toxicity bioassays,
analysis of chemical constituents and bioaccumulation tests in the procedure.
The manual recognizes the dynamic status of ecological evaluation methods
and the need to develop new procedures. The evaluative methods included in
this manual, in conjunction with on-going and recently completed research
studies, will probably form the framework for an evaluative manual covering
the effects of fresh water dredging activities on ecosystems.
The current criteria for the evaluation of the quality of Great Lakes
harbor sediments is based on EPA guidelines (EPA, 1977). These criteria
are based on levels of chemical parameters in sediments (principally total
volatile solids, chemical oxygen demand (COD), total Kjeldahl nitrogen, oil
and grease, mercury, lead and zinc although ammonia, cyanide, phosphorus
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and a number of other metals may be considered), field observations and
benthos. However, projection of bulk sediment chemical analyses to potential
adverse ecological effects due to dredging activities is difficult. The
criteria do not identify the fractions of sediment which are potentially
toxic and biologically available. The problem of bioaccumulation of chemicals
in flora and fauna resulting in magnification in aquatic food chains is not
adequately addressed.
In recognition of the need for a comprehensive program to assess and
predict the environmental aspects of dredging and dredged material disposal
operations, the Corps of Engineers has conducted a five year Dredge Material
Research Program (DMRP) through its Engineers Waterways Experimental Station
(WES) aimed primarily at ocean disposal of dredged spoils. This program
addresses a wide variety of environmental considerations due to dredging in-
cluding the mechanism and the magnitude of chemical constituent release from
dredge material and subsequent effects. Due to the tremendous quantities of
sediment dredged annually and the extreme complexity in assessing environmental
impacts, it is essential to explore a variety of approaches aimed at assessing
and predicting potential effects.
This research attempts to develop procedures designed to assess potential
harmful effects of harbor sediments subject to dredging. In pursuing this
objective, the applicability, ease of duplication and cost effectiveness of
the methods was taken into consideration. Short term bioassay tests using
fish, benthos and plankton were studied for their suitability in screening
sediment quality. These tests utilized sediment from the Duluth-Superior
harbor and Lake Superior and the freshwater species Hexagenia limbata (burrow-
ing fauna), Daphnia magna (planktonic fauna) and Pontoporeia affinis (burrow-
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ing fauna) in 96 hr toxicity determinations. Bluegil1 sunfish (Lepomis
macrochirus) served as the biological probe in cough frequency and breathing
pattern measurements. Information on the usefulness of changes in the activity
of specific enzymes as a tool to monitor sediment quality was developed. The
sediment and water systems used in these tests were chemically characterized
for a large number of parameters. Bioaccumulation of PCBs and p,p'-DDE and
ten metals were studied in Chironomid larvae obtained from harbor sediment
sites and Hexagenia limbata exposed to sediment in the 96 hr tests. The use
of high pressure liquid chromatography to screen sediments for bioaccumuable
organic material was also investigated. Attempts were made to correlate
chemical properties of the sediment and water systems to the tests employing
the biological probes.
The results of this project should aid in identifying methods useful in
assessing the pollution status of harbor sediments and possible adverse eco-
logical effects associated with dredging and disposal of spoils. The develop-
ment of meaningful methods is necessary to avoid ecosystem perturbations and,
at the same time, minimize the cost and delay factors associated with removing
sediments from our waterways and their subsequent deposition.
BIOLOGICAL TESTS TO SCREEN SEDIMENTS
Daphnia Bioassa.ys
Daphnia sp. have been widely used in screening bioassays for a variety of
individual toxicants (Biesinger and Christensen, 1972; Anderson, 1944; Macek
et al., 1976) and complex effluents (Winner, 1976; Arthur et al_., 1975).
Daphnia have been shown to exhibit sensitivity to a varity of chemical species.
The available data on Daphnia responses coupled with their short generation
time and ease of culturing suggests that they are a suitable group for use in
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harbor sediment screening bioassays.
Acute 96 hour toxicity tests using Daphnia magna were conducted in situa-
tions causing exposure of these animals to Lake Superior water in contact
test sediments. Additional 96 hr tests resulted in determining toxic effects
of interstitial water extracted from the sediments, elutriate water prepared
from Lake Superior water-sediment suspensions (EPA/Corps of Engineers, 1977;
Keeley and Engler, 1974), and Lake Superior water used as a sediment extractant
(generated pore water). Elutriate water was prepared by shaking sediment with
Lake Superior water, allowing the sediment to settle for one half hour and
centrifuging the suspension (elutriate particulate phase) to remove most of
the suspended particles. The effect of suspended particles on Daphnia magna
survival was studied by exposure of the animals to mixtures of test sediment
elutriate particulate phase with Lake Superior water.
Hexaqenia Bioassays
Hexagenia sp. possess characteristics highly suitable for use in sediment
bioassay tests. The group has a wide distribution and is sensitive to changes
in sediment and related water quality (Fremling, 1-964; Fremling, 1967).
Eriksen (1963) found Hexagenia nymphs to occupy water within 7 m above the
sediments. The animals ingest mud, organic detritus, bacteria and algae from
above, on or below the sediment surface. Smith and Oseid (1974) found behavioral
changes in Hexagenia limbata occurred at below lethal concentrations of
hydrogen sulfide. The animals were found to be suitable for bioassay tests
with oxygen concentrations down to 2 ppm. Prater and Anderson (1977, 1977a)
have utilized this species in acute toxicity tests of sediments. The burrow-
ing behavior of Hexagenia facilitates direct contact with the sediments thus
promoting uptake of bioavaiTable toxicants associated with the particulates
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and interstitial waters. The relatively large size of the nymphs provides
adequate tissue sample for chemical analysis.
Behavior studies and acute 96 hour toxicity tests were carried out
using Hexagenia limbata in aquaria containing test sediments and overlying
Lake Superior water. The locations of the animals were observed over this
period and their survival was noted at the conclusion of the tests.
Pontoporeia Bioassays
Pontoporeia affinis is the predominate member of the macrobenthic fauna
in all the Great Lakes. Although a burrowing species, it undergoes vertical
migration in the water column, making it a highly versatile sediment quality
probe. Changes in activity have been correlated with bioaccumulation of
mercury from sediments (Magnuson, et_ aj_., 1976). Previous studies demonstrated
that P.. affinis show sediment behavior which may be related to sediment quality
(Marzolf, 1965; Gannon and Beeton, 1969).
Acute 96 hour bioassay tests were conducted involving the exposure of
Pontoporeia affinis to elutriate water-Lake Superior water mixtures in systems
containing sedimented particulates from the test sites as substrates. The
survival of the animals at the conclusion of the tests was recorded.
Fish Behavior Bioassay
Cough response of several species of fish has been used as an effective
indicator of acutely toxic concentrations of individual pollutants (Drummond,
et al., 1973; 1974) and complex effluents (Walden, et_ al_., 1970). Recent
studies of complex effluent screening suggests application of the procedure
using small volumes of test water in 24-48 hr fish exposures (Carlson and
Drummond, 1978). Due to the sensitivity of the method and its potential low
cost, it was tested for its applicability to screening harbor sediment quality.
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In these tests, bluegill sunfish (Lepomis macrochirus) were exposed to Lake
Superior water and to mixtures of Lake Superior water and interstitial water
obtained from harbor sediments for a period of 96 hr. Cough frequency and
change in breathing patterns were measured as end points for these tests.
SYSTEMS EMPLOYED AND CHEMICAL CHARACTERIZATION
The various tests using biological probes to monitor sediment quality
employed both sediment and liquid phase exposure systems. Chemical and physical
analyses of these systems were made for numerous parameters which would char-
acterize the test media and allow chemical correlations to the acute toxicity
bioassay results. The bioassay tests employing fish, benthos and plankton as
probes were studied during the summer of 1977 and 1978= The chemical characteri-
zations of the test sediments and aqueous phases were more extensive during the
sunrner of 1977 than during the summer of 1978.
The exposure systems consisted of sediment, water overlying the sediment,
interstitial water extracted from the sediment under anaerobic conditions,
elutriate water prepared from Lake Superior water mixed with sediment which
was either exposed to air or kept under nitrogen prior to elutriate formation
and Lake Superior water used to extract water solubles or colloids (generated
pore water). During 1978, particulate phase elutriate water was also used
as an animal exposure system.
The sediments were generally analyzed for the "Jensen Criteria" parameters
and additional metal content indicative of the chemical form of the metals
and their biological availability. The sediment metal characterizations were
made according to the amounts in the interstitial water, exchangeable phase,
easily reducible phase, organic and sulfide phase, moderately reducible phase
and residual phase metals (Engler, et al_., 1974; Chen, et al_., 1976). Analyses
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of sediments for specific organic compounds included determinations of some
polychlorinated biphenyls (PCB) and pesticides, some polynuclear aromatic
hydrocarbons (PAH) and pentachlorophenol. The sediments (cores) were also
analyzed for particle size, pH and Eh.
The water phases were generally analyzed for heavy metals, iron, oxygen
content, pH, chemical oxygen demand (COD), NH3, H^S, chloride, suspended solids
and some trace organics. The waters used in certain of the bioassay studies
were analyzed for most of these parameters.
UPTAKE OF CHEMICALS BY CHIRONOMIDS AND HEXAGENIA
Samples of Chironomids were obtained from the sediment sites within the
Duluth-Superior Harbor. Hexagenia limbata exposed to test sediments for 96 hr
in the acute toxicity studies were collected. These biological samples, were
analyzed for some specific organic compounds (PCB, PAH, pesticides) and some
heavy metals. Determination of the accumulation of these chemicals in the
benthic organisms was undertaken to better understand the relationships between
sediment concentrations, bioavailability and bioaccumulation potential. Some
recent studies have shown that the available form of organic chemicals includes
some of the chemical adsorbed to fine particulate matter (Nimmo, ert al_., 1971;
Courtney and Denton, 1976; Zitko, 1974; Nathans and Bechtel, 1976). A number
of metals (primarily methylated forms) accumulate in organisms where they may
lead to adverse effects on the animal or render it unfit for human consumption
(Wood, 1974).
BIOACCUMULATION DETERMINATIONS USING HIGH PRESSURE LIQUID CHROMATOGRAPHY
A measure of the relative tendency of an organic chemical to bioaccumulate
in the lipids of animals is given by its n-octanol/water partition coefficient
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(P). For example, values of log P have been used to predict the bioconcentra-
tion factors of organic chemicals in fish (Neely, et_ al_., 1974). Recent studies
have shown that log P values for a large number of organic chemicals can be
correlated to their retention times on a reverse phase column employing a high
pressure liquid chromatograph (HPLC) with methanol-water mixtures as the mobile
phase (Veith and Austin, 1976; Veith and Morris, 1978). Since it would be
very time consuming and expensive to attempt to identify and quantitate the
complex mixture of organic chemicals in sediments, screening of organic ex-
tracts by HPLC may prove to be a rapid and inexpensive method to evaluate their
potential bioaccumulation hazard.
Organic extracts from harbor sediment sites and a Lake Superior site
were injected into a HPLC system and the eluting chemicals were monitored by
ultraviolet absorption. Similar HPLC separations of extracts from harbor site
Chironomids and Hexagenia limbata were carried out. The extracts were quali-
tatively evaluated for the presence of chemicals with high log P values and a
summary of method feasibility has been presented.
CHEMICAL CORRELATIONS TO BIOASSAYS USING BIOLOGICAL PROBES
The results of the bioassays using animal toxicity and fish behavior as
end points have been analyzed through regression analyses for correlations
with chemical characteristics of the sediments and water systems employed.
Chemical parameters in the various water systems were investigated for toxic
levels when possible. Speculations on the feasibility of using chemical para-
meters pertaining to the sediments and water systems to predict adverse ecolog-
ical effects of dredging have been made.
Sediments from test sites were ranked based on chemical analysis of a
large number of parameters pertaining to the sediments or the interstitial
water extracted from the sediments. The numerical rankings gave an indication
of the relative amounts of the chemical species contained in the sediments or
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-9-
interstitial water. The rankings were tested for correlations to the animal
survival results in order to determine whether or not sediment and/or inter-
stitial water chemistry were predictors of toxic effects in the bioassay
systems.
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-10-
SECTION 5
STUDY AREA
GENERAL HARBOR DESCRIPTION
The Superior-Duluth harbor includes over 11,500 acres of which 3,100 acres
is dredged channel, 22-27 feet deep; 650 acres is natural river channel, 6-15
feet deep and; 7,750 acres is breakwater bays and sloughs, less than 6 feet
deep (DeVore 1978). The area is characteristic of most estuaries being his-
torically shallow and possessing critical habitat for shore birds, waterfowl
and fish in addition to industrial development and shipping.
The estuary is located at the southwest tip of Lake Superior and is
separated from the main lake by the largest natural freshwater bay-mouth sand
bars in the world; Wisconsin and Minnesota Points (Figure 1). A natural
opening through the bars to the lake was maintained in the early years by the
currents of the Nemadji and St. Louis Rivers at a location which is now the
Wisconsin entry (Figure 1).
The harbor was geologically formed by the combination of declining water
levels and a subsequent rise of the north side of the lake basin. Declining
water levels resulted in construction of a series of three parallel sets of
sand spits (beaches). The last two sets of these bars divide the harbor from
the lake (Wisconsin and Minnesota Points) and the outer harbor from the inner
harbor (Connors and Rices Points). A third and older set form the base of the
Grassy Point and Arrowhead bridges (Figure 1). Subsequent to the water level
decline, rebound of the northshore from the weight of glacial ice has resulted
in flooding of the St. Louis and Nemadji River mouths and the low lying areas
between the sand spits. The effect of this natural change was formuation of
these bays which make up the harbor. Superior and Allouez Bays were formed
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FIGURE: 1
SLTERIOR-DULUTH HARBOR AREA.
ZONE III
:one I
ZONE (IV
;one r
O \ZONE V
it-
zone VI
ZONE
SUPERIOR
ZONE VIII
-Wisconsin
Entry
ti*
W'
-------
by inundation of the low land between the two most recently formed sets of _ j
sand bars, St. Louis Bay which makes up the inner harbor was formed by
flooding of the St. Louis River mouth and low lands between the inner two sets
of sand bars.
The partial separation of Allouez and Superior Bays (Section 6, Figure 2)
undoubtly results from sediments deposited by the Nemadji River. The Nemadji
flows through highly erodable unconsolidated red-clay sediments which were de-
posited during high water stages of the lake. The red turbid water of the
Nemadji deposits much of its sediment load in the harbor, the natural entry
and the lake proper.
The red water and sediment deposited in the harbor made navigation diffi-
cult during the early settlement years. To solve this problem piers were con-
structed in the natural entry to narrow and deepen the channel and dredging was
initiated in-the 1860's. Since that time, over 55 million cubic yards of sedi-
ment has been dredged to form the extensive network of channels, slips, docks
and a second entry; the Minnesota entry located on the northwest end of the
harbor. In addition to being responsible for development of deep water areas,
deposition of spoil is responsible for development of all the major islands in
the lower harbor (Barkers, Hog and Herders) and the formation of some shoal
water areas. Over the years a large percentage of the spoil has been deposited
in the open lake (Corps of Engineers 1974).
Establishment of navigation channels was stimulated by mining of Mesabe
Range ore deposits, the harvest of native white pine forest stands and wheat
farming in the plains states. The improvements in navigation channels re-
sulted in rapid industrial development along the shoreline and the growth
of the Superior, Wisconsin-Duluth, Minnesota metropolitan area, the largest
on Lake Superior. Iron ore or iron concentrate (taconite) has remained
the primary commodity shipped from the port since the early years. Montana
coal and grain represent the second and third most important cargos
shipped in recent years. Salt, limestone, petroleum products,
-------
cement, fish and steel are also shipped through the port. Total shipments
during the period from 1967-1972 ranged between 37-43 million tons. Con-
struction of a major coal shipment facility in 1974 has resulted in a
substantial increase in shipments of coal.
HISTORY OF POLLUTION
Water quality, which is often considered a modern problem, declined
in the Superior-Duluth estuary during the early period of development.
Newspaper accounts describing pollution of the St, Louis River indicate
sludge mats and methane gas odors occurred in the upper harbor to Cloquet
as early as 1914 and resulted from Kraft mill effluents and saw mill wastes.
Water quality monitoring was begun by the city of Duluth in 1946 and showed
mid-summer dissolved oxygen below the minimum requirements for most fish
species during the first year of record. Reduced oxygen undoubtly resulted
from the decay of wood fiber mats, municipal sewage (which was not treated
by either city prior to 1950) and ship discharges of sewage.
Harbor zones were identified in the 1972-1973 impact assessment program
according to forces and sources of contamination which influence them
(National Biocentrics Inc. 1973). Zone I near the Duluth, Minnesota entry
(Figure 1) is influenced by Lake Superior storm wave activity, vessel move-
ment and storm runoff from the Duluth sewage system (National Biocentrics
Inc. 1973). The western edge of this zone and zone II, the Duluth turning
basin, are also influenced by grain dust which introduces the potential
pesticide contamination and by other industrial developments. Zone III
represents a shallow backwater area in the inner harbor previously used
for dredged spoil disposal which also received discharges from the Duluth
municipal treatment facility.
The St. Louis River represents the dominant force in the inner harbor
area represented by Zone IV. In addition to paper mill effluents and sedi-
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-13
merits carried by the river, grain elevators, coal docks, ore docks, and a
Minnesota Power and Light plant are located in the inner harbor and re-
present potential sources of contaminants. Fly ash deposits have been
plowed into the harbor by the power station and represent one of the most
obvious potential hazard areas in this zone.
Zones V, VI and VII include a deep water channel area (Zone V) in-
fluenced primarily by ship movement; a shallow area (Zone VI) which his-
torically has received storm and direct sanitary sewage discharges from the
Superior, Wisconsin treatment plant and; a shallow zone (Zone VII) bordered
by a relatively undeveloped shoreline. The area near and including the
Superior entry (Zone VIII) is influenced by Lake Superior seiches, major
sediment discharges from the Nemadji River and ship movement into ore and
ore concentrate loading facilities located in the area.
PHYSICAL NATURE OF THE SEDIMENTS
No quantitive information on physical or chemical quality of harbor
sediments was available prior to 1970 except for engineering data obtained
from sediment cores taken prior to channel modification work^ and at con-
struction sites. However, during the period between 1970-1977 extensive
sampling was conducted to define the physical-chemical nature of harbor
and western Lake Superior sediments. Harbor data were collected by the
U.S. Environmental Protection Agency (15 samples, 1970; 6 samples 1973;
33 samples 1975; 6 samples 1976) National Biocentrics Incorporated (16
samples; 1972-1973) the University of Wisconsin-Superior (36 samples,
1973) the University of Wisconsin-Madison (6 samples and the U.S. Army
Corps of Engineers (60 samples, 1977). An additional 13 sites were analyzed
by the University of Wisconsin-Madison.
1 Corps of Engineers, Boring Logs for the Superior-Duluth harbor 1958-1961.
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-14-
Sediment borings taken during 1958-1961 by the U.S. Corps of Engineers
from the inner harbor and Allouez Bay (areas not previously dredged) show
the sediments below the surface to be inorganic silts and clays which are
underlaid by silty sands in some areas. These borings and surface samples
(power dredge, National Biocentrics Inc., 1973) show the upper layers in
the inner harbor and Superior entry area to be composed of poorly sorted
mixtures of clay and silt. Sieve analysis of these sediments showed the
inner harbor and Superior entry area (which are influenced by the St.
Louis and Nemadji Rivers) are comprised of over 451 silt-clay sized par-
ticles whereas, sediment from other areas of the harbor seldom contained
over 20% silt and clay sized particles.
Comparisons of harbor and lake sediments made by Van Tassel and Moore
(1976) showed that harbor sediments in general contained a higher percentage
of clay sized particles and were not as well sorted as lake sediments.
The lake sediment studies indicate that currents remove finer particles
from inshore areas, and transport them to deep water zones. The general
pattern results in removal of fine sediments from the Wisconsin inshore
area and deposition in deeper offshore Minnesota waters.
CHEMICAL NATURE OF THE SEDIMENTS
Chemical analysis performed during 1970-1973 were conducted to define
the quality of harbor sediments with respect to EPA guidelines (EPA
criteria as given in Appendix D) for volatile solids, chemical oxygen demand,
total Kjeldahl Nitrogen, total phosphorus, oil and grease, mercury, lead,
and zinc. In addition to the above, arsenic, cadmium and copper were measured
during most investigations. The chemical analysis demonstrated that some samples
in every zone exceeded the guidelines for most parameters and that the chemical
nature of the sediments was extremely variable within and among harbor
-------
zones (National Biocentrics Inc., 1973).
Correlations analysis showed that heavy metal and oil and grease
concentrations were directly related to the percentages of clay and silt
sized particles in the sediments, supporting the hypothesis that fine
particles provide more binding sites and combine with metals and organic
compounds in the sediments (National Biocentrics Inc., 1973). As part of
subsequent study by UW-Madison, Helmke et_ al_. (1977) measured the con-
centrations of 22 metals in harbor and lake sediments and found then to be
3 to 4 fold higher in the clay sized (<2 u) fractions. This analysis of
undisturbed red clay soil particles of the region demonstrated that the
natural ranges of zinc and copper exceeded the 1973 EPA guidelines in the
absence of pollution. Native soils contain between 140-230 mg/kg zinc
and 65-88 mg/kg copper (Helmke et_ al_. 1977).
None of the samples collected during the autumn of 1973 by UW-Madison
(a total of 6) or subsequently by the US EPA during 1975, 1976 contained
the concentration of mercury identified through the 1970 EPA or 1972-1973
National Biocentrics and UW-Superior analysis (National Biocentrics Inc.,
1973). Although mercury concentration in bulk lake sediments did not exceed
0.24 mg/kg mercury,clay sized fractions contained as much as 4.22 mg/kg
mercury. Comparisons were made between concentrations of 22 metals in
standard red clay soils and clay sized sediments from harbor and lake sedi-
ments, For both harbor and lake sediments, ratios calculated by dividing
the concentration of copper, zinc, chromium, mercury and arsenic in sedi-
ments by the concentrations in native soils exceeded one whereas, ratios
calculated for metals not utilized by man were equal to one. The larger
ratios show that harbor and lake sediments have been altered by man.
More recent measurements of contaminants in sediments have been
directed at defining concentrations of toxic organic compounds particularly,
polychlorinated biphenyls (PCBs). A limited survey conducted by EPA during
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-16-
1976 showed pesticide concentrations were generally low but concentrations
of PCBs were high in two of eight samples (exceeded 30 mg/kg; EPA,1976).
A more extensive survey conducted by the US Corps of Engineers during
1977 (Whiting, 1977) included 30 sampling sites and showed PCB concentra-
tions did not exceed 0,5 mg/kg in harbor sediments and generally did not
reach 0.1 mg/kg (dry weight).
AQUATIC LIFE
The potential for promoting human health problems or damage to the
aquatic life community in the estuary and western end of Lake Superior,
represent primary concerns in dredging and dredge spoil disposal. Some
measurements on the aquatic life community have been performed to define
the potential for the above problems and to determine the pollution status
of harbor sediments. Identification of macroinvertebrates and measurement
of species diversity suggests the bottom community is dominated by pollution
tolerant forms in most zones. Surveys conducted by UW-Superior (National
Biocentrics Inc., 1973) and the US EPA (EPA 1975) indicate that the presence
of intolerant forms and high species diversity are limited to zones directly
influenced by lake seiches. Abundance and distributions of macroinverte-
brates have been shown to be positively correlated with the percentage of
clay and silt sized particles in harbor sediments, which chemical analysis
demonstrates contain the highest concentration of toxic metals (National
Biocentrics Inc., 1973; Helmke et al_., 1977). The question of whether zinc,
or mercury in sediments are available to aquatic life in the harbor and
western end of the lake has been studied by Helmke et al_. (1976). Employing
both chemical techniques which measure exchangeable and soluble zinc in
sediments and comparisons between concentrations of zinc in sediments and
in aquatic life sampled at various sites, Helmke et_ al_. (1976) found zinc
in sediments is not available to macroinvertebrates and does not appear
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-17-
to bioaccumulate in the aquatic food chain. Their limited analysis indi-
cated that mercury in sediments may be available to macroinvertebrates and
does bioaccumulate in the aquatic food chain. Analysis of mercury levels
in sediments, macroinvertebrates (Pontoporeia affinis), sculpins and some
other fish (white fish, lake trout and burbot) suggested the highest con-
centrations occur in animals at the top of the food chain. Upon exposure
to available soluble forms of mercury and zinc, Magnuson et_ al_. (1976)
found behavioral changes occurred in Pontoporeia affinis resulting in
increased susceptability of the animals to predators (sculpins).
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-18-
SECTION 6
PROJECT FIELD SAMPLING
SAMPLING SITES
Samples were collected from six site areas in the Superior-Duluth
Harbor during this research project. The sites were selected to provide
a diversity of bottom sediment types and to allow comparison to previous
sediment data. Reference samples were collected from the Pokegama Bay
during 1977 and from Lake Superior in 1978. These reference samples were
used as the controls in all bioassays (Section 8). Sampling locations
are shown in Figure 2 and include:
1. Allouez Bay - Highly organic and clay type sediments are found in
this region. Samples were collected from the northern edge of
the eastern end of the dredged channel at a water depth of ap-
proximately 25 feet.
2. Superior Ore Dock Area - A clay-sand bottom dominates this area.
Collection of samples at this site occurred approximately 50
yards northeast of the Northern Pacific ore dock at a depth of
about 25 feet.
3. Lakehead Transshipment Terminal Area - The sediments were fairly
high in volatile solids indicating substantial organic material.
Sampling was done about 30 yards north of the end of the Lakehead
Slip in approximately 24 feet of water.
4. Superior Municipal Sewage Treatment Plant - Organic sediments of
domestic sewage origin were abundant. Samples were collected east
of the treatment plant at the eastern edge of the shio channel in
about 28 feet of water.
4a. Superior Municipal Sewage Treatment Plant - Sampling was done
east of the treatment plant at the western edge of the ship channel
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-19-
at a depth of approximately 20 feet.
5. Cargill Inc. Elevator B - Sediments were a mixture of sand and
organic material. Sampling of this site was located approximately
150 yards north of the end of the Cargill Elevator B slip in
about 24 feet of water.
5a. Farmers Union Elevator - Sediment in this area contained organic
matter of industrial origin. Samples were collected west of the
end of the Farmers Union slip at a water depth of approximately
5 feet.
6. Minnesota Power and Light Company Generating Station - Fine sedi-
ments with substantial amounts of fly ash material dominated this
region. This sampling location was approximately 700 yards east -
northeast of the power plant in about 19 feet of water.
P. Pokegama Bay - Organic sediment minimally affected by human
development occurred in this area. Sampling of this reference
site was done at the southern end of the west branch of Pokegama
Bay. Samples were collected in about 4 feet of water.
L.S. Lake Superior - The sediment at this location contained substantial
amounts of eroded silt and clay. Collection of samples occurred
approximately 6 miles north of the Superior Entry to the Duluth-
Superior Harbor at a water depth of about 95 feet. The Lake
Suoerior site replaced Pokegama Bay as the reference site in 1978
because chemical analysis of Pokegama Bay sediments showed high
concentrations of chemical oxygen demand and mercury.
SEDIMENT, WATER AND CHIR0N0MIDS SAMPLING
Samples of sediment and water from the Duluth-Superior harbor area
and the Lake Superior site were collected using the University of Wisconsin-
Superior RV GULL. Each sampling site was located according to shoreline
structure references and water depth. During 1977, seven cores were ob-
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-20-
tained at each location employing a Benthos type 217 gravity corercontaining
cellulose acetate butyrate liners 4 foot (1.2 m) in length and 2.5 inches
(6.3 cm) inside diameter. Upon obtaining a core, the ends of the core
liner were stoppered and it was stored upright in an insulated box type
holder cooled with ice. The core liners contained about 8 to 18 inches
(20.3 to 45.7 cm) of harbor sediment with the remainder of the liner filled
with overlying harbor water. The cores were utilized in chemical analyses
and bioassay tests (usually the day following sampling).
During 1977, portions of the overlying water from several of the cores
were siphoned into glass or polyethylene containers for later use in chemical
analysis and enzyme activity tests. Preservatives were added to sub-samples
of the overlying water and subsequently analyzed for ammonia, total sulfide
and total phenols. The sub-samples were cooled in an ice chest.
Sediment samples were also obtained at each site employing a Ponar
dredge. Portions of these sediment samples were transferred to glass
bioassay chambers for tests using Daphnia magna placed in water above the
sediments and using the burrowing fauna Hexagenia limbata. These bioassay
tests commenced the day following this sampling.
Portions of the sediment were rinsed through a 35 mesh size stainless
steel sieve and the Chironomids retained by the sieve were placed in a
glass vial containing harbor water. A second method consisted of hand
mixing sediment with harbor water in a metal wash basin and collecting the
animals appearing on the water surface. The Chi ronomids were cooled in an
ice chest and transported to the laboratory for weighing and freezing
prior to chemical analysis.
During 1978, a similar sampling program was carried out for harbor
sites except that water overlying the cores was not analyzed and a fewer
number of cores were obtained at each site. Dredge sediment was used from
only two of the harbor sites and the Lake Superior site for Hexagenia
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-21-
1imbata toxicity tests.
The reference site used in bioassay tests during 1977 was the Pokegama
Bay slough. Core samples were obtained by inserting the cellulose butyrate
acetate core liner into the sediments at a water depth of 4 feet (1.2 m).
During 1978, the Lake Superior site was used as the reference site in bioassay
tests.
The sediment and water samples were collected on the dates listed in
Table 1.
HEXAGENIA LIMBATA
Two locations were used during the study for collecting Hexagenia
1imbata. Animals used in toxicity tests conducted during 1977 were col-
lected from a bay on Rainy Lake in northern Minnesota (International Falls,
Minnesota). The second source of Hexagenia 1imbata was Ox Creek in northern
Wisconsin. The Ox Creek Hexagenia were used in 1978 bioassay and bio-
accumulation studies.
The collection procedure consisted of obtaining the upper layer of
sediment with a shovel, transferring the sediment to screen boxes and
rinsing the sediment with water from the collecting location. The animals
were removed from the screen box as they were exposed using plastic spoons
and placed in water in a cooler. During transport to the laboratory, the
water was aerated and kept at about 15 C. Sediment was collected from the sampling
location for use in storing the animals prior to their use in the tests.
PQNTQPOREIA AFFINIS
The Pontoporeia were collected from Lake Superior at a 75 foot (29.9 m)
water depth at a location north of the Superior entry to the Superior
Harbor. The location was approximately two miles from the entry.
The upper layer of sediment was obtained with a Ponar dredge and
placed in wash basins. Water was added and mixed with the sediment to
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-22-
TABLE 1. Sediment and Water Sample Collection Dates
1977 Sampling
Site Date Collected Reference Site Date Collected
1 June 12 Pokegama June 12
2 June 5 Pokegama June 5
3 July 10 Pokegama July 10
3R October 14 Lake Superior October 12
4 July 17 Pokegama July 17
4R October 21 . Lake Superior October 12
5a July 24 Pokegama . July 24
6 June 26 Pokegama June 26
6R August 14 Pokegama August 14
L.S. August 7 Pokegama August 7
Pokegama June 19 ----
1978 Sampling
1 June 2 Lake Superior June 2
2 June 23 Lake Superior June 17
2 July 14 Lake Superior July 6
3 June 17 Lake Superior June 17
4 June 8 Lake Superior June 2
4a July 14 Lake Superior July 6
5 June 23 Lake Superior June 17
5 July 7 Lake Superior July 6
5 July 21 Lake Superior July 20
6 July 7 Lake Superior July 6
6 July 21 Lake Superior July 20
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-23-
fortn a slurry. The slurry was poured through a screen box Into a second
basin containing lake water. The animals were floated away from the screen,
collected with a plastic spoon and placed in a four liter plastic bottle
containing lake water. The container was placed in a cooler and transported
to the laboratory for storage,
LEPQMIS MACROCHIRUS
The bluegills (Lepomis macrochirus) used in the fish cough re-
sponse bioassays were collected from Bass Lake, a small lake located in
Bayfield County near Delta, Wisconsin.
The fish were obtained by seining the inshore areas of the lake.
Only those bluegills that were between 7 and 11 cm in length were retained.
The captured fish were placed in 30 gallon metal containers (lined with
polyethylene) that had previously been filled with lake water. During
transportation to the laboratory the water was aerated and kept in the
dark to minimize temperature changes.
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SECTION 7
CHEMICAL CHARACTERIZATION OF SAMPLES AND
WATER-SEDIMENT SYSTEM PREPARATION
SEDIMENT-WATER SYSTEMS UTILIZED
The sediment-water systems chosen for chemical characterization primarily
consisted of the same systems subsequently used in behavioral response and
acute toxicity bioassay tests. The systems analyzed were sediment, water
overlying the sediment, sediment interstitial water, elutriate water and
"generated pore water". The specific chemical parameters investigated
were chosen to provide a comprehensive survey (within budget and time limita-
tions) of those chemical species indicative of the overall quality of the
systems. •
TREATMENT OF CORE SAMPLES
Background
The procedures utilized in obtaining the various sediment-water systems
from the core samples were designed to maintain sample integrity and to provide
sufficient amounts for chemical and biological tests.
A number of methods for extracting interstitial water from sediments have
been employed by investigators (Presley et al_., 1967; Duchart et_ al_., 1973;
Robbins and Gustinus, 1976). The exposure of sediments to air, temperature
fluctuations and mechanical perturbations can alter the chemical nature of
the interstitial water (Bray e_t a_k , 1973; Troop et al_.» 1974; Bischoff et
al., 1970). For collecting large volumes of interstitial water, the method
used in this investigation involved high speed centrifugation of the sedi-
ment at 4°C followed by collection of the liberated water under a nitrogen
atmosphere. After the initial separation of the water from the sediment,
the water was recentrifuged, decanted and utilized in chemical and biological
tests. The water was not filtered and consequently it contained fine
suspended solids. The interstitial water, containing this fine suspended
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-25-
material, is considered representative of material released to the overlying water
in the event of dredging operations.
Elutriate water was prepared from sediment mixed with Lake Superior water ac-
cording to published procedures (EPA/Corps of Engineers, 1977). However the elu-
triate water was not filtered to avoid possible loss of toxic gases to the air and
possible loss of metals or organics on the filter material. The liquid phase was
obtained by using only high speed centrifugation as the means of separating sus-
pended solids from the aqueous phase. Two elutriate water systems were prepared
for each sediment site (summer of 1977). One system used sediment exposed to air
while the second system employed sediment which had been kept under nitrogen prior
to elutriate water preparation.
An attempt was made to arrive at an estimate of the amounts of chemical species
which could potentially be flushed out of the sediments under vigorous mixing con-
ditions. Sediment (unexposed to air) was repeatly extracted with Lake Superior
water over a period of time to obtain "generated pore water". The chemical char-
acteristics of the generated pore water gives a measure of the potential availability
of chemical species to the water phase.
Procedure
During the summer of 1977, the sediment core samples were extruded and used to
prepare the various sediment-water systems usually on the day following sampling.
The cores had been stored upright in the plastic liners under lowered temperature
conditions (<12 C) in the dark.
The cores were treated by the scheme shown in Figure 3. Nearly all of the
overlying water was siphoned from the top of the sediments and discarded (overly-
ing water had been collected at the time of sampling). Measurements of pH and Eh
were made 2 cm below the surface. The stopper was removed from the bottom of the
core and replaced with one which tightly fit inside the liner. The upper portion of
the liner was placed in a glove bag and the stopper and core were forced toward the
top of the liner. The residual overlying water and upper 1 cm of the core was dis-
carded.
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extruded portion of sediment
remove overlying water
extruded under I
into centri :uge
bottles
extrude sec
into glass
tainer
iment
con-
Organics (Tables 11,12,13,14
PCB, PAH, Pesticide and
Pentachlorophenol Analysis
residue
through~
screen
Sieve
10
un«er
tut
centrifuge
Nk
pH, Eh of
sediment
(Table 10)
,Metal Analysis
Sediment Core Samples
P04, Suspended Solids, CI,
Metals
(Table 15)
(Table 2)
Mercury, COD,
b^S, Chloride,
NH^, TKN, Total P
Total Solids,
Total Volatile
solids, oil &
grease, moisture
total phenols,
particle size
Sediment
for VPI
Enzyme
Work
Form Anoxic
Elutriate
Water
Form Oxic
Elutriate
Water
Cool
(Section 8)
96 hr toxicity
tests, cough
response tests
(Tables 18,19)
NH
Metals
Cool
iCool
Interstitial
water
(Section 8)
96 hr toxicity
tests, cough
response tests
Form
Generated
Pore Water
(Table 16)
pH, COD, H2Ss
CI, NH3, TKN,
PCBs, Suspended
Solids, Metal?
(Tables 4,5,6,
7,8)
Metal Analysis
(Tables 22,23
24,25,26,27,28
COD, PCBs
Metals
Water for
VP I Enzyme
Work
i
ro
cn
a
Figure 3. General Treatment of 1977 Superior-Duluth Harbor Core Samples
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Sediment from the core was transferred to 250 ml stainless steel centri-
fuge bottles using a glazed ceramic spoon. Each cup was filled within about
three-fourths of its capacity (about 180 g of sediment). One core was used
to fill two or three centrifuge bottles. The bottles were capped, removed from
the glove bag ana eentrifuged for 12 minutes at 10,000 rpm and 4°C (IEC model
B-20A refrigerated centrifuge). The centrifuging provided a force of 10,000 x
g at the lower end of the bottle decreasing to 3,500 x g at the upper end.
The cups were returned.to the glove bag where the caps were removed and
the interstitial water decanted into teflon bottles. Since a substantial
amount of particulate matter was present in the water, it was placed in 250 ml
polycarbonate centrifuge bottles and capped. The particulate-water suspension
was recentrifuged at 14,000 rpm (25,000 x g at the lower end of the bottle
progressing to 8000 x g at the upper end) for 15 minutes. The resulting in-
terstitial water was decanted into teflon bottles (exposed to air) and stored
in a refrigerator. After removal of interstitial water, portions of the sedi-
ment from several cores were transferred under nitrogen to a plastic bottle for
subsequent metal analysis.
Other portions of the sediment were removed from cores and placed in two
glass bottles. One bottle was used for general chemical sediment analyses
while the second was sent under an N2 atmosphere to Virginia Polytechnic Institute
and State University, Blacksburg, Va (along with a subsample of the intersitial
water) for use in enzyme activity tests. Another sample (about 1000 g) was placed
on aluminum foil, air dried, and used in organic chemical analysis. Other portions
of the sediment were saved for preparation of elutriate water.
SEDIMENT ANALYSIS
The sediment samples obtained during the summer of 1977 were subjected to
extensive inorganic and organic analysis. This analysis included a number
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-28-
of general physical and chemical tests, specific metal determinations, PCB,
pentachlorophenol and polycyclic aromatic hydrocarbon measurements. A much
more limited sediment analysis was carried out on sediments sampled during
the summer of 1978.
General Physical and Chemical Sediment Tests
Sediment samples obtained from several cores for a site were
prepared for general analysis using EPA procedures (Fuller, 1969). The
sediment was passed through a 10 mesh (approximately 2 mm pore size) sieve
and collected in a large beaker. It was either blended or stirred (in the
case of sediments with high water content) and divided into subsamples.
The subsamples were analyzed for chemical oxygen demand (COD), ammonia,
Kjeldahl nitrogen, moisture content, volatile solids, total phosphorus, total
phenols, oil and grease and total sulfide using EPA methods (Fuller, 1969).
These procedures are summarized in Appendix A.
Particle size analyses were performed by the pi pet-sedimentation
method described by Royse (1970). Total and organic mercury were determined
by the procedure of Olson et_ a]_. (1975). This method involved sediment
digestion in a concentrated mixture of HNO^ and H^SO^, oxidation of organic
matter with KMnO^, reduction of mercury with stannous sulfate followed by
atomic absorption measurements.
The sediment samples obtained during the summer of 1978 were analyzed
only for pH, Eh and particle size. The results of these general sediment
analyses are given in Tables 2 and 3.
Preparation of Sediment Extracts for Metal Analysis
The characterization of the sediments for concentrations of numerous
metals was carried out using a procedure which generally followed the
selective extractive methods of Engler et_ al_. (1974) and Chen et_ a]_. (1976).
The selective extraction of metals results in their classification as to
availability to various chemical extractants.
-------
TABLE 2
GENERAL SEDIMENT ANALYSIS-1977
SITE
Parameter* 1 2 3 4 5a 6 6R** LS Pokegama
t
pH
6.3
6.1
6.2
6.6
6.3
6.5
6.9
7.1
6.
£1^1* (mv)
90
85
60
80
45
55
75
110
80
COD
99,000
104,000
35,000
31,000
kO CO
—< en
» •
o o
o o
o o
136,000
138,000
57,000
62,000
133,000
168,000
34,000
168,000
Ammonia
46
79
5.3
500
720
1900
1400
500
620
140
139
580
690
1840
1230
107
167
TKN
660
675
270
205
3000
4300
7200
7400
1700
2400
1200
1200
4400
4000
5600
4900
129
220
Total S
26
43
2.4
3.1
1.8
1.6
240
220
89
82
87
67
300
210
11 .2
6.2
23
21
Total P
130
75
75
83
1600
820
1400
1050
110
1700
1400
1500
2000
78
Oil and Grease
1700
1700
2200
4300
9600
12300
3700
3500
7200
2900
4500
3500
5600
4800
1120
880
Total Phenols
2.0
0.2
2.2
2,9
1.7
1.5
1.8
1.6
2.0
1.2
0.8
2.5
3.2
1 J
Organic N
610
600
265
200
2500
3600
5300
6000
1200
1750
1060
1110
3800
3300
3700
3600
22
53
Total Mercury
0.66
0.08
0.64
0.77
0.13
0.23
0.23
0.04
0.!
* Values are in mg/kg of dry sediment except for pH, Eh.
**R denotes a repeat sampling of site 6.
Obtained about 2 cm below surface.
-------
TABLE 2. (continued)
GENERAL SEDIMENT ANALYSIS-1977
Parameter
1
2
3
S ITI
4
:
5a
6
6R
LS
Pokeg.
Total Solids {%)
35.7
61.6
45.2
36.6
63.6
51.6
43,5
45.0
45.0
Total Volatile
Solids {%)
11.2
5.1
Q 4
11.9
6.4
9.7
11.1
4.7
12.5
% Sand
4.2
37.2
16.0
5.1
58.6
30.5
20.5
9.0
4.7
(>62.5y)
5.5
15,6
4.3
57.4
36.5
17.0
10.5
4.1
% Silt
36.4
42.9
39.1
45.0
23.4
45.7
46.4
55.2
36.8
(3.9-62,5p)
35.8
37.8
40.7
21.0
31.6
cc q
CO A
o6 A
"3Q Q
JO . O
% Coarse Clay
19,0
6.8
17.8
21.4
7.9
13.3
17.2
12,2
39.8
(0.98-3.9y)
18.5
16.2
18.3
7,2
21.5
I A -»
1 mm ¦ l
13.5
21.7
% Fine Clay
40.3
9.7
27.1
28,5
10.2
10.5
15.9
23.7
18.5
(<0.98y)
40.2
30.4
36.6
13.0
10.3
14.4
23.6
35.4
-------
TABLE 3
GENERAL SEDIMENT ANALYSIS-1978
SITE
Parameter 1 2 2R 3 4 4a 5 6 L.S.
pH*
6.5
--
6.3
6.4
6.5
6.5
--
6.3
6.5
Eh* (mv)
101
--
104
yg
36
38
__
-49
113
Total Solids {%)
/I
%Jf (— * nr
52.1
—
40.0
38.7
~
61.5
50.4
--
Total PCBs**
0.31
0.61
--
1.6
1 .0
--
0.85
0.77
0.33
% Sand
13.5
2.6
3.5
2.6
62.2
4.4
1.12
±2.0
±0.5
±2.9
±2.5
+4 # "j
±3.8
±0.2
% Silt
40.7
40.7
32.6
23.7
22.1
53.7
72.7
±3.6
±6.5
±11.4
•1 A
± 1 U • 0
±1.0
±2.7
% Course Clay
14.8
±1.2
17.2
±2^4
--
±3.5
24.8
±0.2
—
c c
o. b
±1.0
11.7
7.4
±0.2
% Fine Clay
31 .0
39.5
42.4
49.9
10.0
30.7
L. I « G
±2.9
±4.7
±10.7
±13.0
±4.3
±0.9
* Obtained about 2 cm below surface.
**Expressed as mg/kg of dry sediment.
-------
-32-
Sediment subsamples were obtained under a nitrogen atmosphere from the
upper layers of sediment in the centrifuge bottles from which the interstitial
water had been removed. Total metal content analyses of sediment from two of
the sites were carried out on uneentrifuged sediment still containing inter-
stitial water. The classification of metals according to their extractability
and procedures employed are summarized below.
Exchangeable Phase Metals --
The exchangeable metals are obtained by extraction with ammonium
acetate. This treatment removes metals which are largely adsorbed to
mineral and organic surfaces. The metals in this phase are believed to
be in rapid equilibrium with metals in the interstitial water phase. They
are available to replenish metals which are removed biologically and chemi-
cally from the interstitial water.
150 ml of 1.0 M deaerated ammonium acetate solution (deaerated by
bubbling through the solution) was added (under a atmosphere), to
about 22 to 30 g dry weight of the centrifuged sediment from which the
interstitial water had been extracted. The sediment was put into seal able
centrifuge cups. Since the sediment was not completely dry, part of the
sediment was used to determine the percent dry weight of the sample. The
sealed plastic centrifuge bottle was shaken for 90 minutes with a mechanical
shaker. After shaking, the exchangeable phase was separated by centrifuga-
tion followed by filtration through 0.45 micron filters. Duplicate samples
were usually run through the entire metal extraction procedure.
Easily Reducible Phase Metals --
The metals in the easily reducible phase are obtained by using hydro-
xy 1 amine hydrochloride and a dilute solution of nitric acid as the extractants.
This treatment removes metals which are mainly associated with hydrous
-------
-33
oxides of manganese. Four to five gram (dry weight) subsamples were taken
from the residue to the previous extraction (exchangeable phase) and washed
with deionized water. A portion was used to determine the dry weight of
the sample. The washed residue was extracted with 200 ml of 0.1 M hydro-
xylamine hydrochloride in 0.01 M HN03 solution by shaking the mixture for
45 minutes with a mechanical shaker. The extract was separated by centri-
fuging and filtering the supernatant. The aqueous phase was acidified to
0.2% HN03 for metals analysis.
Organic and Sulfide Phase Metals --
Treatment of residue from the easily reducible phase with concentrated
hydrogen peroxide and acidic ammonium acetate releases metals which are
associated with metal sulfides ana the organic fraction of the sediment.
Also manganese from MnO^ not extracted as part of the easily reducible phase
is obtained. The organically complexed metals vary in stability and bio-
availability but may be subject to release from the sediments by microbial
degradation.
The residue from the easily reducible phase was washed with deionized
water. A subsample of 1 to 2 g (dry weight) was weighed into a plastic
centrifuge cup and treated with 3 ml of 0.02 M HNO-j and 5 ml of acidified
30% H202 (acidifed to about pH 2). A portion of the initial washed residue
was used in determining the dry weight of the sample. The mixture was
heated in a water bath at 85°C for 5 hours. After 2 hours of heating,
another 2 to 3 mis of acidified 30% H^O^ was added. After the mixture was
allowed to cool to room temperature, 25 ml of 1.0 M ammonium acetate in
6% HN0- was added and the sample shaken in a mechanical shaker for 30 minutes.
Residual solids were separated by centrifugation and filteration.
Moderately Reducible Phase Metals --
Extraction of the residue from the previous step with hydroxy!amine
-------
-34-
hydrochloride in acetic acid liberates reduced iron and metals associated
with hydrous iron oxides. These metals could be released from the sediments
to the overlying water under reducing conditions.
The entire residue from the previous step was washed with deionized
water and treated with 40 ml of 0.04 M hydroxy1 amine hydrochloride in 25%
acetic acid. The resulting mixture was shaken well , heated at 100°C for
3 hours in a water bath. After 3 hours, the mixture was cooled to room
temperature and the extract separated as in previous steps.
Residual Phase Metals —
The moderately reducible phase residue contains weathered minerals
which is generally the location of the largest amounts of metals in the
sediment. The metals are in the inner layer positions of clay minerals
or within the mineral crystal lattice. These metals are stable and in a
biologically unavailable form.
The residue from the previous extraction was washed with distilled
water (which was discarded) and a 0.5 to 0.8 g (dry weight) portion was
weighed into a 50 ml teflon beaker. (A portion was used to determine the
dry weight in the sample.) The sample was digested with a mixture of 10 ml
concentrated HF, 5 ml concentrated HNO^, and 1 ml concentrated HCIO^ in a
heated sand bath at about 170°C (surface temperature of sand), and evaporated
to near dryness. The residue was dissolved in 5 ml concentrated HC1 (heat-
ing was usually necessary). If some undissolved residue remained, about
20 ml deionized water was added and the solution was filtered through a 0.45
micron filter. The filtrate was stored in a plastic bottle. (If after
the original digestion, and upon addition of the HC1, the residue completely
dissolved, then deionized water was added and the sample was brought to a
volume of 50 ml. At this point the sample was ready to be analyzed for metals.)
The undissolved residue was scraped and washed off the filter into a
platinum evaporating dish. The platinum evaporating dish, containing the
-------
-35-'
residue and wash water, was dried overnight at 100°C, cooled, and weighed.
Enough sodium carbonate was added to completely cover the dry residue (1-
5 ratio) and the dish and its contents were placed in a muffle furnace at
about 900°C. As the Na2C03 melted, the dish was swirled to insure complete
mixing of residue and Na^CC^. The platinum dish was removed from the
furnace and allowed to cool to room temperature. When cool, 1 ml of deioniz-
ed water was added and the mixture heated slowly on a hot plate at low set-
ting. Concentrated HC1 was carefully added until the sample was acidic.
The mixture was stirred and heated on the hot plate until all the solid
dissolved. At this time, the filtrate from the original digestion was
added and the combined solution evaporated to near dryness. Finally,
deionized water was added to bring the solution to a 50 ml volume.
Metal Analysis Procedures
Atomic absorption spectroscopy was used in the metal analysis. Flame
absorption methods (employing direct aspiration of the aqueous phase extracts)
were used for those samples containing relatively high metal concentrations
(generally 0.1 to 1.0 ppm or above in the extracts). For those extracts with
metal concentrations below those detectable by direct aspiration into the flame,
a graphite furnace was used. The instruments utilized were the Perkin Elmer
Model 306 atomic absorption system and the HGA 2100 graphite furnace with back-
ground correction. Pyrolyzed graphite tubes were used to increase detection
limits (Manning and Ediger, 1976).
Metal Analysis Results
The concentrations of metals found in the various phases of the sediment
are given in Tables 4 through 8. These values are computed on a dry weight basis.
The concentrations are corrected for any weight losses occurring during the ex-
traction steps and for reagent blanks.
-------
TABLE 4
Metal Concentrations in Exchangeable Phase of Sediment Samples (mg/kg)-1977
As Cd Cr Co Cu Fe Pb Mn Ni Se Zn
Site
1
<0.08
<0.40
<4,1
<1
6
<0.40
170
<1 .6
170
<1.6
<0.16
<0.40
Si te
1
<0.08
<0.39
<3.9
<1
5
<0.39
170
<1.5
110
<1.5
<0.15
<0,39
Site
2
<0.04
<0.17
<1.7
<0
7
<0.17
3.6
<0.7
105
<0.7
<0.07
<0.17
Site
2
<0.06
<0.27
<2.7
<1
1
<0.27
—
<1.1
171
<1.1
<0.11
<0.27
Site
3
<0.05
<0.23
<2.3
<0
9
<0.23
32.2
<0.9
61.9
<0.9
<0.09
<0.23
Site
3
<0.05
<0.26
<2.6
<1
0
<0.26
29.2
<1.0
48.2
<1.0
<0.10
<0.26
Si te
4
<0.05
<0.26
<2.6
<1
0
<0.26
*1.0
<1.0
90.7
<1.0
0.10
<0.26
Site
4
<0.06
<0.31
<3.1
<1
2
<0.31
1.2
<1.2
133
<1.2
<0.12
<0.31
Site
5a
<0.05
<0.22
<2.2
<0
q
<0.22
20.7
<0.9
A-jf rt
a f Vil
<0.9
<0.09
<0.22
Site
5a
<0.05
<0.23
<2.3
<0
9
<0.23
11.7
<0.9
31.5
<0.9
<0.09
<0.23
Si te
6
<0.08
<0.41
<4.1
<1
C
0
<0.41
7.8
<1.6
96.7
<1.6
<0.16
<0.41
Site
6
<0.08
<0.40
<4.0
<1
6
<0.40
9.1
<1 .6
93.7
<1.6
<0.16
<0.40
Site
6R
<0.04
<0.19
<1.9
<0
7
<0.19
4.0
<0.7
100
<0.7
<0.07
<0.19
Site
6R
<0.04
<0.21
<2.1
<0
8
<0.21
4.5
<0.8
124
<0.8
<0.08
<0.21
Site
LS
<0.07
<0.33
<3.3
<1
3
<0.33
17.8
<1.3
65.8
<1.3
<0.13
<0.33
Site
LS
<0.06
<0.29
<2.9
<1
2!
<0.29
11.1
<1.2
59.5
<1.2
<0.12
<0.29
i
to
OFi
8
-------
TABLE
5
Metal
Concentrations in Easily Reducible Phase of Sediment Samples
(mg/kg)~l977
As
Cd
Cr
Co
Cu
Fe
Pb
Mn
Ni
Co
JC
Zn
Site
1
<0
45
<2.2
<22
<8.9
<2.2
840
<8.9
97
<8.9
<0.89
<2.2
Site
1
<0
4c
%m)T
<2.2
<22
<8.9
<2.2
660
<8.9
76
<8.9
<0.89
<2.2
Si te
2
<0
36
<1.8
<18
<7.2
<1.8
420
<7.2
94
<7.2
<0.72
<1.8
Site
2
<0
26
<1.3
<13
<5.2
<1.3
270
<5.2
99
<5.2
<0.52
<1 .3
Site
3
<0
41
<2.1
<21
<8.2
<2.1
1400
<8.2
110
<8.2
<0.82
3.3
Site
3
<0
48
<2.4
<24
<9.6
<2.4
1400
<9,6
77
<9.6
<0.96
3.9
Site
4
A
-------
TARI F fi
I /%»>*# Imw Im
Metal Concentrations in Organic Phase of Sediment Samples (mg/kg)-1977
As
Cd
Cr
Co
Cu
Fe
Pb
Mn
Ni
Se
Zn
Si te
1
*
<0.79
<7,9
3.1
18.1
880
22.0
165
5.5
26.7
Site
1
2.4
<0.67
<6.7
3.1
17 J
860
21 5
139
<2.7
<0.27
23.8
Si te
2
1.8
<0.98
<9.8
3,9
14.4
690
3 9
93.6
<3 9
<0.39
25 2
Site
2
2.2
<1.0
<10.0
<4.0
19.5
990
8.0
84.4
<4,0
0.80
1 Q T
lo. 1
Site
3
4,4
1.1
<7.9
4.7
OQ Q
0 ¦/ • ¦/
4600
53.8
85.9
4,9
0.47
87.8
Qi j-n
| \^\m»
3
4.2
6.3
<8.8
7.0
49.9
5300
68.3
103
69.3
0.53
155
Site
4
3.8
1.4
<8.0
4.8
47.8
6700
83.4
155
17.2
0.32
152
Si te
4
5.6
1.1
<8.0
6.4
42.8
9500
84.9
155
8.1
0.64
133
Site
5a
2.5
<0.95
<9.5
<3.8
15.2
1900
41.8
69.7
8.2
<0.38
31.3
Site
5a
2.3
0.78
<7.8
5.1
29.6
3400
63.9
89.1
2.5
<0.31
75.0
Site
6
3.2
1 .1
<7.1
6.6
"j g g
900
41.0
84.7
404
0.28
88.1
Si te
6
5.2
0.75
<7.5
7.1
23.3
1000
50.8
102
209
0.30
88.5
Site
6R
5.1
1.6
<9.1
4.9
30.1
6600
77.4
134
22.1
0.36
72.0
Site
6R
9.8
1.7
<8.7
4.7
33.5
6400
91.4
133
41»9
Q gj
178
Site
LS
1.8
<0.82
<8.2
5.4
21.2
3900
6.5
73.2
127
<0.33
53.8
Site
LS
2.2
<0.69
<6.9
4.6
20.7
4000
5 5
73.9
15 9
0.28
19.6
-------
TABLE 7
Metal Concentrations in Moderately Reducible Phase Sediment Samples (mg/kg)-1977
As Cd Cr Co Cu Fe Pb Mn Ni Se Zn
Site
1
<0.20
<1 .0
<10.1
5.4
10.8
2500
<4.1
62.9
8.7
<0.41
22.7
Site
1
0.34
<0.87
<8.7
4.0
8.8
1400
<3.5
60.8
8.7
<0.35
20.0
Site
2
<0.18
<0.91
<9.1
5.4
9.2
1700
<3.6
61 J
6.4
<0.36
11.4
Si te
2
0.32
<0.82
<8.2
6.0
14.6
1600
<3.3
57.2
8.8
<0.33
14.9
Site
3
0.20
<1 .02
<10.2
<4.1
9.2
2900
<4.1
48.8
4 Q
i *
<0.41
19.1
Site
3
0.68
<1.1
<11.2
8.3
11.9
4000
<4.5
63.0
10.3
<0.45
29.9
Site
4
0.38
<0.95
<9.5
<3.8
10.6
4700
<3.8
60.9
c; q
J , j
<0.38
32.7
Site
4
0.38
<0.95
<9.5
<3.8
10.6
4900
<3.8
62.8
5.1
<0.38
30.4
Site
5a
1.02
<1.3
<12.7
5.1
8.8
4700
<5.1
61.1
14.8
<0.51
34.6
Site
5a
0.63
<1.0
<10.4
<4.2
7.9
4600
<4.2
62.5
10.4
<0.42
40.6
Site
6
0.37
<0.93
<9.3
4.3
6.0
3000
<3.7
53.8
59.3
<0.37
31.7
Site
6
0.78
<0.98
<9.8
3.9
6.4
2300
<3.9
60.8
70.6
<0.39
33.6
Site
6R
<0.24
<1.2
<11 .9
6.5
8.2
8400
<4.8
117
12.9
<0.48
51.7
Si te
6R
0.48
O
< 1 .Z
<11 .9
4.8
12.7
7600
<4.8
90.9
7 4
/ « ™
<0.48
41.4
Si te
LS
0.63
-------
TABLE 8
Metal Concentration in Residual Phase of Sediment Samples (mg/kg)-1977
As Cd Cr Co Cu Fe Pb Mn Ni Se Zn
Site
-j*
2.6
<4 4
^ T • 1
<43.9
17.6
!7.1
8,800
<17.6
149
23.7
<1.8
35.1
Site
^ *
4.9
<4.9
<48.8
00 C
CC , D
26.7
• 9,400
<19.5
186
26.4
<2.0
47.9
Site
2*
5.3
<4,4
<43.9
20.2
14.9
18,100
<17.6
167
<17.6
<1.8
34.3 ,
Site
3.3
<4.1
<41.4
24.8
18.3
17,600
<16.6
166
. 1 c c.
< ID .0
<1.7
34.8
Site
3
24.8
6.2
176
70.3
57.5
46,100
51.7
393
59.9
32.1
57,9
Site
3
25.4
8.5
182
61.8
70.5
41,300
48.5
376
52.1
29.1
55.7
Site
4
24.1
8.6
<43.1
44.0
30.1
38,900
<17.2
285
43.1
13.8
42.3
Site
4
18.7
4.3
<42.5
45.9
16.6
42,000
<17.0
272
145
11.1
40.8
Site
5a
22.4
<6.6
<65.9
<26.4
23.7
31,100
<26.4
316
<26.4
5.3
42.2
Site
5a
10.9
9.1
<45.6
24.6
42.9
39,300
<18.2
252
<18.2
21.9
58.4
Site
6*
7.3
<3.7
132
42,5
72.9
11,500
36.7
198
4R h
Tv a w
13.2
51.3
Site
6*
12.2
9.3
137
76.2
34.4
11,300
50.3
208
63.3
2.2
67„6
Site
6R
15.4
<4.0
<40.5
25.9
28.3
47,100
20.2
373
<16.2
2.4
80.2
Site
6R
13,1
<4.4
<43.6
43.6
57.6
47,200
<17.5
355
<17.5
8.7
r* f\ n
59,4
Site
LS
28.4
<4.9
<48.9
36.2
35.3
42,000
<19.6
438
34.3
19.6
69.5
Site
LS
19.4
5.1
<51.1
<42.9
30.6
39,100
<20.4
434
T-Jt
<20.4
10.2
68.4
^Samples were not carried through the Na^CO^ fusion step.
1
O
r
-------
-41-
The sediment samples analyzed by this selective extractive method were
obtained from the upper layers of centrifuged samples. This material is en-
riched in smaller particles which would settle in water at the slowest rate
after suspension due to a disturbance of the sediment. Consequently, it should
contribute a more important environmental effect (compared to the total sedi-
ment) when subjected to dredging. Therefore the upper layer of the residue
(obtained after removal of the interstitial water) was utilized in the selec-
tive extractive procedure.
The enrichment of metals in the upper layer of this residue is illustrated
by analysis of total metal content (by digestion in Parr bombs as described
below) of centrifuged and uncentrifuged sediment for two sites. These re-
sults for sites 4 and LS are presented in Table 9. It is observed that the
upper layer of the centrifuged sample has total metal concentrations about two
times higher than those found in the bulk uncentrifuged sample.
Analysis for Total Metals in Sediments
Separate sediment samples from each of the sites were digested in Parr
bombs with aqua regia and HF for total metal content determinations (Bernas,
1968). These sediment samples were not centrifuged but were sieved through a
#10 mesh stainless steel sieve to remove coarse particles and debris from the
sediments. These analyses were conducted to allow comparisons to the EPA
pollution status classifications (Appendix D) currently in use for Great Lakes
harbors (EPA, 1977). These results are listed in Table 10.
Pesticide and PCB Analysis
Sediment samples were analyzed for selected chlorinated pesticides and
polychlorinated biphenyls (PCBs). These chemical compounds affect sediment
quality since they are potentially available to biota in the ecosystem. Many
-------
TABLE 9
Total Metal Concentrations in Sediment Samples (mg/kg)-1977
As Cd C r Co Cu Fe Pb Mn Ni Se Zn
S *i t €¦ 4
37
<10
<31
43
39
46,900
92
670
65
<4.1
310
11 Site 4
36
<11
<32
34
40
46,800
96
700
49
<4.3
320
ySite LS
29
<10
<31
44
17
35,600
<31
560
67
<4.2
82
^Site LS
36
<13
<38
41
20
35,600
<38
590
<46
<5.1
76
cSite 4
38
<24
<70.
<75
J*
70,800
212
1040
<85
<9,4
CCA
DDu
cSite 4
30
<25
<76
<81
a a
OD
75,800
1270
1010
207
<10.1
520
cSite LS
C *5
•}
<16
<47
94
69
68,800
<47
1090
72
<6.3
178
cSite LS
83
<15
<46
80
67
67,500
<46
1100
71
<6.1
178
^Uncentrifuged Samples
cCentrifuged Samples
-------
TABLE 10
Total Metal
Concentrations
in Sediment Samples
(mg/kg) -
Uncentrifuged Samples
-1977
As
Cd
Cr
Co
Cu
Fe
Pb
Mn
Ni
Se
Zn
Site
1
—
<12
41
50
24
52,600
60
980
180
151
Site
1
-~
<10
67
33
40
48,100
52
900
155
<4,2
1 DO
Site
3
64
<13
<38
it*|h
13
38,500
90
360
<46
<5.1
202
Site
3
66
<12
<35
A A
<38
12
37,600
82
490
61
4.7
190
Site
4
28
<15
<46
80
31
49,500
. 77
710
105
<6.2
275
Site
4
31
<16
<47
66
31
50,000
109
720
56
<6.3
278
Site
5a
38
<12
- <35
38
<12
25,900
59
380
42
<4»7
111
Site
5a
19
<12
<36
39
<12
26,700
61
390
<44
<4.9
114
Si te
6
28
<13
<38
66
13
40,800
590
148
<5.1
214
Site
6
25
<12
<37
51
12
41,700
86
560
142
<4.9
199
Si te
6R
97
<10
<30
43
20
42,500
91
550
85
<4.0
221
Site
6R
27
<11
<33
<36
18
42,400
78
540
lob
<4,5
214
Si te
LS
9
<11
<34
<36
18
36,200
57
520
95
<4.5
88
Si te
LS
39
<11
<33
35
22
39,500
55
550
57
<4.4
88
i
-ps»
GO
9
-------
-44-
chlorinated hydrocarbon type organic compounds concentrate In the fatty tissue
of aquatic animals according to their solubility characteristics.
Sediment Extraction Procedure ~
The extraction of pesticides and PCBs from sediment samples generally
followed published procedures (Breidenbach e£ al_., 1964; Bellar and Lichten-
berg, 1975). Each sediment was air dried (about 3 days) and a 40 g sub-
sample was mixed with 4 ml distilled water and transferred to a pre-ex-
tracted cellulose soxhlet extraction thimble. The sediment was extracted
with a 9:1 mixture of hexane-acetonefor 8 hours in a soxhlet extraction
system. The extract was passed through a column of ^SO^ and concentrated
to about 5 ml using a Kuderna-Danish evaporator system equipped with a
three ball Snyder reflux column.
Extract Clean-up Procedure ~
For removal of lipids, waxes and high molecular weight colored com-
pounds, the extract was passed through a column containing a lower layer of
3% deactivated florisil and an upper layer of ^SO^. Elution was done
with a 70:30 mixture of petroleum ether and methylene chloride. The eluent
was concentrated to 5 ml using the Kuderna-Danish apparatus. Recovery
efficiency was determined to be 86% for PCBs and about 100% for DDT.
The concentrated eluent was subjected to further clean-up for removal
of high molecular weight interferences and elemental sulfur using gel
permeation chromatography (Kuehl and Leonard, 1978). The extract was in-
jected into the system and pumped (FMI metering pump) through two 2.5 cm
diameter glass columns containing a total of about 60 g of SX-2 Bio-Beads
(BIO-RAD Laboratories). Methylene chloride was used as the mobile phase
solvent. The U.V. absorbance of the eluent was monitored at 254 nm. A
standard mixture of corn oil and PCBs was used to determine the point of
-------
-45-
separation between the high and low molecular weight eluents. Elemental
sulfur eluted at a retention volume greater than found for PCBs (and other
compounds under analysis) and this late eluting fraction was discarded.
The eluent fraction containing the low molecular components was concentrated
to 10 ml using a gentle stream of pre-purified nitrogen blown over the sur-
face.
Separation of PCBs and Pesticides —
The separation of PCBs from chlorinated pesticides in the gel permeation
cleaned-up extract was accomplished using column chromatography with silica
gel as the stationary phase (Snyder and Reinert, 1971). Silica gel was
heated at 210°C overnight and covered with pentane. A portion of the
silica gel saturated with pentane (5 ml) was transferred to a 10 ml graduated
pi pet filled with-pentane. After rinsing the silica gel column with pentane,
2 ml of the gel permeation cleaned-up extract was placed on the column and
eluted with 70 ml of pentane. This eluent contained the majority of the
PCB components. Benzene (40 ml) was passed through the column to obtain
chlorinated pesticides and the remaining PCB compounds. Spiked samples
carried through this procedure showed 84% of the PCBs were contained in the
pentane fraction and 100% of the DDT was present in the benzene fraction.
Gas Chromatographic Analysis --
The pentane and benzene fractions from the silica gel separation pro-
cedure were concentrated by applying a stream of pre-purified nitrogen
over the liquid surface.
General gas chromatographic analysis procedures followed EPA methods
(Thompson, 1977). The samples were analyzed using a Tracor MT-220 gas
63
chromatograph equipped with Ni electron capture detectors. Two columns
were used in the identification and quantitation process. One column con-
tained 4% SE-301/6% 0V-210 on Chrom W (HP) 80/100 mesh while the second
-------
-46-
column was packed with 1.5% OV-17/1.95% 0V-210 on Chrom W (HP) 80/100
mesh (Analabs Co.).
The gas chromatograph elution patterns were analyzed for the presence of
lindane, aldrin, chlordane, pp'-DDD, pp'-DDE, op'-DDT, pp'-DDT, dieldrin, en-
drin, heptachlor, heptachlor epoxide, hexachlorobenzene, methoxychlor, oxychlor-
dane, and PCB Aroclor mixtures 1019, 1221, 1232, 1242, 1248, 1254 and 1260. The
details of the elution pattern analysis for PCBs are given in Appendix B. The
results of these chemical analyses are given in Tables 3 (for 1978) and 11 (for 1977).
The concentrations of lindane, heptachlor, aldrin, heptachlor epoxide,
dieldrin and the DDT complex were less than 1.0 pg/kg in all the sediment sam-
ples. The total PCB values ranged from about 0.3 to 2.1 mg/kg. For the 1977
summer sediment samples, the chromatographic elution patterns showed best fits
with Aroclor 1242 and Aroclor 1254 mixtures. Low levels of p,p'-DDE were found
in two of the sediment samples.
Chlorinated Phenol Analysis
Chlorinated phenols are present in certain industrial effluents such as
paper processing wastes. Pentachlorophenol has a variety of uses among which
include its function as a wood preservative. In order to determine if sediments
in the Duluth-Superior harbor were contaminated with pentachlorophenol, an
analytical survey was conducted.
Analysis Procedure —
About 100 g of dried sediment was placed in a glass bottle and 500 ml
of 0.1 M NaOH solution added. The suspension was shaken for 0.5 hr and
centrifuged for 15 minutes at 10,000 rpm. The liquid was decanted into a
separatory funnel and extracted with 100 ml of 85:15 hexane-methylene
chloride. The organic"layer was discarded. The aqueous layer was acidified
to pH = 2 with 6M HC1 and extracted with two 50 ml portions of 85:15 hexane-
methylene chloride. After a final extraction with 50 ml of hexane, the
-------
TABLE 11
*
PCBs, Pesticides and Pentachlorophenol in Sediments - 1977
Silica Gel Samples
Total Total a &
Site** PCBs PCBs* 1254ff 1242ff p,p' DOE Pentachlorophenol
1
1.2
1.4
0.42
0.36
<2
1.4
2
0.54
Not Separated
<2
1.0
3
1.4
1.6
0.72
0.24
<2
2.7
4-S
1.9
2.1
0.73
0.39
<2
15
4-F
0.9
0.89
0.31
0.19
9.0
5a-S
0.72
0.48
0.1,5
0.07
<2
36
5a-F
0.93
1.0
0.23
0.34
<2
6
1.7
1.4
0.4
0.37
<2
4.3
6R
0.98
1.1
0.31
0.33
3.1
10
LS-S
0.66
0.49
0.35
0.12
<2
1.2
LS-F
0.38
0.12
No Pattern
Discernable
*PCB values given in mg/kg dry weight of sediment; values are not corrected
for spike recovery adjustments (65% for total procedure). DDE and pen-
tachlorophenol concentrations are ng/g dry weight of sediment.
**S indicates sample from summer collection period.
F indicates sample from special fall collection period.
"'"These values were determined from the sample extracts after florisil and gel
permeation chromatography clean-up steps by calculating the concentrations
for each PCB component and summing the values for all the components to
give the total concentration.
tfotal values were determined from the combination of the PCB values in the
pentane and benzene fractions after silica gel separation. Total concentra-
tion calculations were determined with the same method used in #3 above.
The 1254 and 1242 values were determined with the aid of a computer program
to be the best fit with the possible PCB standards and the sediment extracts.
(Appendix B)
-------
-48-
combined organic layers were filtered through anhydrous Na2S0^ and reduced
to about 10 ml in a Kuderna-Danish evaporator system.
A portion of the sediment extract was treated with diazoethane as an
ethylating reagent. The diazoethane was generated from N-ethyl-N'-nitro-N-
nitrosoguanidine in an aqueous NaOH solution-hexane mixture. The extract
was treated with the hexane saturated with diazoethane. Upon completion
of the reaction, a solution of 80:20 methanol-H^O was added to remove by-
products of the reaction. The hexane layer was separated and concentrated
with a stream of N2 (Rogers and Keith, 1976).
The concentrated hexane extract was analyzed by gas chromatography
using the same equipment as described for pesticide and PCB analysis. The
results of this analysis is included in Table 11.
Pentachlorophenol was detected in most of the 1977 sediment samples.
However the concentrations were low being in the range of 1 to 15 yg/kg.
PAH Analysis
Polycyclic aromatic hydrocarbons (PAH) can enter ecosystems from fossil fuels
and the combusion of fossil fuels. A number of compounds under the general PAH
classification are carcinogenic (Panel on Polycyclic Organic Matter, 1972). The
sediments were examined to assess possible contamination from PAH' s.
A1though no information exists on the occurrence of specific PAH compounds
in sediments from the Duluth-Superior harbor or Lake Superior, several PAH's
have been reported in Great Lakes water or sediment (Strosher and Hodgson, 1975).
The compounds, 2-methylanthracene, 9-methylanthracene, benz[a]anthracene, benzoCa]-
pyrene, and 3-methylcholanthrene, which contain from three to five aromatic rings,
were used in technique development, including the determination of extraction ef-
ficiency and analytical sensitivity. Five additional compounds (phenanthrene,
1 -methylphenanthrene, perylene, benzo[£,Jh,i_]perylene and dibenz[a_,h]anthracene)
reported from water or sediment samples (Strosher and Hodgson, 1975; Acheson ert
al., 1976) were also included as targets in the initial sediment screening
-------
-49-
procedure. In the quantitation stage of the analysis, an investigation was also
made of the occurrence of fluoranthene, pyrene and anthracene in the sediments.
Analysis Procedure--
Portions of the extracts (hexane-acetone extractions florisil and gel per-
meation clean-up, and silica gel fractionation) used in the PCB analysis were
investigated for PAH compounds. For preliminary screening, the extracts were
chromatographed on a SE-30 column using a Varian Aerograph HY-FI Model 600D gas
chromatograph with a flame ionization detector. The instrument was operated
isothermally at 185-190°C for elution of 2-methylanthracene, 9-methylanthracene,
phenanthrene, and 1-methylphenanthrene and isothermally at 265-270°C for elution
of benz[a]anthracene, benzo[a]pyrene, 3-methylcholanthrene, perylene, benzo[g_,h_,i_]-
perylene and dibenz[a_,h]anthracene. Red clay soil was spiked with five PAH com-
pounds and tested for extraction recoveries. The results showed recoveries of
52.4% for 3-methylcholanthrene, 72.5% for benzoCa]pyrene, 66.0% for benzCaJanthracene,
76.8% for 9-methylanthracene and 90.2% for 2-methylanthracene.
None of the PAH compounds under investigation were observed by preliminary
screening in the sediment extracts from sites 1, 2 and 3 (Superior Harbor area)
or Lake Superior. Elution peaks were noted in the extracts (benzene fraction
from silica gel separation) from sites 4 (Superior Harbor), 5, 6 (Duluth Harbor
area) and Pokegama Bay.
Further work on identification, and quantitation of compounds was performed
at the Environmental Research Laboratory, Duluth, The sediment extracts from
sites 4, 6, Pokegama Bay and Lake Superior were analyzed by combined gas chromato-
graph-mass spectrometry methods (GC/MS). The procedure involved separation of
sample components using a Finnigan 9610 GC equipped with a flame ionization
detector. Compound separation took place on a column containing 3% 0V-101
on Gas-Chrom Q in an oven programmed from 100°C (1 minute) increasing by 4°C/min
to a final holding temperature of 225°C. A more detailed description is provided
in Appendix B.
-------
-50-
Analytical Results--
Several PAH compounds were identified by mass spectrometry in the extracts
from sites 4, 6 and Pokegama Bay. GC/MS retention times, molecular weights,
formulae, and approximate concentrations for the identified compounds are
given in Tables 12, 13 and 14. When more than one compound is listed at
a given molecular weight, the order of listing is in decreasing order of identity
probability. The determination of identity possibility and the order of probability
was determined from a computerized library search of approximately 25,000 compounds
(Heller and Milne, 1978). Gas chromatograms for the sediment extracts from the
three sites and Lake Superior are given in Appendix B.
PAH's identified from the GC/MS analysis of the sediment extract from site
6 were basically the same as those from site 4, with the exception of the com-
pound that produced the second or later eluting signal at MW 252 from site 4
being absent in site 6. Retention times between compounds of the same MW's
at the two sites are in good agreement.
phenanthrene was identified by GC/MS from the Pokegama Bay sediment
extract. The mass spectral library search listed 2,4,5,7-tetramethylphenan-
threne, 3,4,5,6-tetramethylphenanthrene, and 1-methyl-7-(l-methyl ethyl)-
phenanthrene as spectra matches with the unknown at MW 234. The compound at
MW 234 produced the greatest ion intensity (870) of the PAH compounds. An
unknown compound of MW 168 or 169 with a long retention time (47:34 min.) was
also present in Pokegama Bay sediment. The library search listed 1,11-bi phenyl -
4-amine, 1,1'-biphenyl-3-amine, and 1,1'-biphenyl, 3-methyl as the three
possibilities based on similarities of mass spectra. Of these three, 1,1'-
biphenyl, 3 methyl was least likely from
-------
Table 12. PAH Compounds from Site 4 Sediment
Approximate
Retention time Concentration*
(min,) MW Formula Compound (ppm, dry wt.)
11:30 178 C-.4H-1Q phenanthrene 0.72**
anthracene
13:26 192 C^H^ methyl phenanthrene 0.32
15:52 202 C]gH10 fl uoranthene 1.24
16:32 202 ClgH1() pyrene 0.81
18:24 234 ^8H18 C^-phenanthrene ---
21:16 228 C,QH10 benzfalanthracene 0.97
18 12 triphenylene
chrysene —
26:00 252 ^20H12 benzo[k]fluoranthene —
benzo[e)pyrene —
perylene 0.38
27:24 252 C2QH12 benzo[a]pyrene 0.81
benz [e Jacephenanthrylene
benzo"["k]f1 uoranthene —
~Concentrations are not corrected for recoveries through analytical process.
**Where more than one compound is listed at a given MW, concentrations are
calculated based on the compound indicated.
-------
-52-
Table I3. PAH Compounds from Site 6 Sediment
Retention time
(fflin.)
11:28
178
Formula
C14H10
Compound
phenanthrene
anthracene
Approximate
Concentration*
(ppm5 dry wt.)
0,72**
13:48
15:50
16:34
18:20
21 :16
192
202
202
234
228
C15H12
C16H10
C16H10
C18H18
C18H12
methylphenanthrene
fluoranthene
pyrene
C^-phenanthrene
benz[a]anthracene
chrysene
triphenylene
0.27
1.27
0.85
1.06
26:02
252
C20H12
benzo[k]f1uoranthene
benz[ejacephenanthry1ene
benzoOJf 1 uoranthene
*,**Same as in Table 1 2.
Table 14. PAH Compounds from Pokegama Bay Sediment
Approximate
Retention time Concentration*
(min.) MW Formula Compound (ppm, dry wt.)
11:24 178 ci4Hio phenanthrene** <0.06
18:26 234 ^18^18 C^-phenanthrene —
21:18 228 C]8H12 benz[a]anthracene* <0.03
47:34 169 ^12^11^ unknown
~Concentrations are not corrected for recoveries through analytical
process.
**Due to weak ion intensity signals, library searches were not used;
tentative identifications are made from molecular weights and reten-
tion times.
-------
examination of the spectra, as there was no evidence of methyl group loss.
The long retention time of this unknown would indicate a high boiling point,
possibly higher than the biphenyl-amines
Weak ion intensity signals at MWs 178 and 228 at retention times of
11:24 and 21:18 indicated the presence of phenanthrene (anthracene) and
benz[a]anthracene (chrysene, triphenylene), respectively. No methyl -
phenanthrene, fluoranthene, pyrene, or heavier PAH's were identified from
Pokegama Bay as they were from sites 4 and 6.
Discussion of Sediment Results
General Sediment Parameters —
The general sediment parameters given in Tables 2 and 3 indicate high
levels of certain constituents. COD values were above the 80,000 mg/kg
level for sites 1, 3, 4, 6 and Pokegama Bay. A value above 80,000 is in
the heavily polluted category according to current Great Lakes harbor
criteria (see Appendix D). Ammonia and TKN values were high for sites
3, 4, 5a, 6 and LS. The TKN values classify these sediments as heavily
polluted for this parameter. These same sites exhibited elevated levels
of oil and grease.
The sediments all showed positive redox potentials except for one
sample (site 6-1978). These Eh values were measured on core samples at a
distance of about 2 cm below the sediment surface. Mercury levels were
below the 1.0 mg/kg standard for all the sediment samples and thus they
would be classified as non-polluted with respect to this parameter.
A general consideration of all of these sediment parameters would
indicate that site 2 was the least polluted with sites 1 and LS showing
low levels of some values (see Table 45, Section 10).
The relationship of these general sediment parameters to potential ad-
verse ecological effects due to their disturbance and disposal is not well
understood. For example, a COD value will include a measurement of all
material which can be oxidized by a fairly strong chemical oxidizing agent
-------
-54
under rigorous conditions. Consequently ammonia and iron (II) would be
measured in addition to oxidizable organic matter. The rate and degree of
oxygen demand by sediments can vary greatly for the same COD values depend-
ing on the species contributing to the COD's.
The use of oil and grease values in determining sediment quality have
been questioned (DiSalvo, et^ al_., 1977). The oil and grease value can con-
tain numerous natural and petroleum derived hydrocarbons in addition to
fats, oil, waxes, and sulfur. Sediments with high oil and grease values
showed low toxicity toward mussels, crabs, clams and snails in their tests.
Metals in the Sediments --
Total metal concentrations in Table 10 indicate elevated values for
certain metals in the Duluth-Superior harbor sediments. The results show
high levels of lead, iron, nickel and arsenic and moderate to high levels
of manganese and zinc in the harbor sediments when compared to classifica-
tions of sediment metal levels (Appendix D). These total metal concentra-
tions were obtained by completely dissolving the sediment samples in Parr
bombs with aqua regia and HF. Thus they are expected to be higher than those
obtained by using a concentrated HNO^, 30% HgOg digestion procedure (Fuller,
1969). This latter procedure gives metal concentrations in the sediments
compatible with values listed in EPA guidelines but does not include metals
within the silicate lattices in the particles. Consequently our total metal
values cannot be directly compared to EPA guideline values.
The classification of the sediment metals according to their release
characteristics to various chemical extractants provides information on
the nature of their bioavailability (Tables 3 through 8). Within the detec-
tion limits of the analyses, only iron and manganese were extracted from
the sediments in the exchangeable and easily reducible phases. However,
many metals were associated with the organic and sulfide phase of the sedi-
ment. As shown in Table 6, detectable amounts of arsenic, cobalt, copper,
-------
-55-
nickel and zinc were found in the organic and sulfide phases as were selenium
and cadmium in some of the samples. The metals in this phase are potentially
available to biota in the ecosystem (Engler et a]_., 1974), The amounts of
nickel and zinc in some of the samples were greater in the organic and sul-
fide phases than in any of the other phases.
The metal concentrations in the moderately reducible phase of the
sediments were generally lower than in the organic and sulfide phases.
However, cobalt and nickel showed similar concentrations to those found
in the organic and sulfide phase. Detectable amounts of arsenic, copper
and zinc were found in the moderately reducible phase and large amounts of
iron were present.
The residual phase of the sediments contained detectable levels of
all investigated metals. Compared to the other phases, arsenic, cobalt,
iron, nickel and selenium were found in the highest amounts in the residual
phase. The amounts of copper and zinc were similar to that found in the
organic and sulfide phase. The amount of lead in the residual phase was
less than determined in the organic and sulfide phase. Manganese was dis-
tributed throughout the separate phases with the largest amounts detected in
the residual and organic and sulfide phases of the sediments.
Excluding the metals in the residual phase of the sediments and summing
the concentrations of metals in the other phases indicates that the non-
residual (and potentially bioavaiTable) metal levels were generally not
high. All of these values would fall below the heavily polluted category
with the exception of nickel for four sites (particularly site 6 which
may have been contaminated) and lead for three sites.
Trace Organics in the Sediments --
The concentrations of PCBs in the sediment samples were not high.
Tables 3 and 11 indicate a total PCB range of 0.3 to 2.1 mg./kg. The samples
obtained at the same site during two different years show good agreement.
-------
-56-
The values determined in this study are higher by about a factor of ten
compared to results from a recent Corps of Engineers harbor survey (Whiting,
1977 and are below the 10 mg/kg upper limit for unpolluted sediments (Appendix D).
The sediment data indicate a general absence of chlorinated hydrocarbon
pesticide compounds in the sediments. The concentrations of pentachlorophenol
were low (1 to 36 ng/g).
The concentrations of the PAH compounds identified in sediments from
two sites (4 and 6) were about 1 mg/kg or less. There are no sediment quality
standards for these compounds. At these low levels, it is very unlikely that
any adverse ecological effects would result from their presence in the sedi-
ments. Of the compounds tentatively identified, benz[a_]anthracene is carcin-
ogenic and benzoCa3pyrene is highly carcinogenic.
OVERLYING WATER
Samples of water overlying the sediments were obtained by siphoning a por-
tion of the water contained in the core liners. These samples were analyzed for
a number of physical and chemical parameters indicative of overall water quality.
Specific procedures used in the analysis are given in Appendix A and the results
are presented in Table 15. In addition to these data, measurements of dissolved
oxygen, temperature and specific conductance were made in the field during sampling.
The results of these field measurements are given in Appendix C.
The water samples were not filtered and consequently the results do include
contributions from suspended particulate matter. It is likely that the metals
are largely associated with fine particles.
There are generally no large differences in parameters between sites. Site
2 overlying water is seen to be particularly low in nitrogen values. The Duluth
harbor water exhibits higher specific conductance values than found for Superior
harbor water (Appendix C).
-------
TABLE 15
CHEMISTRY OF WATER
OVERLYING
SEDIMENT -
1977
C T T C
X f fcfWI
Parameter*
1
2
3
4
5a
6
6R
LS
pH
7.5
6.7
6.3
6.8
7.2
—
7.0
7.5
Total PCBs
0.04
—
0.2
0.04
0.03
0.2
b^S
0.17
0.20
0.20
<0.17
<0.19
0.19
0.20
—
<0.17
<0.18
<0.17
<0.17
NH~
j
0.90
0.05
3.7
3.4
4.3
4.2
2.2
3.8
1.6
TKN
7.0
0.05
5.9
7.8
10.5
12
9.7
16
14.5
COD
36.5
« o
V w « (m.
32.2
60.1
60.1
64.2
50.1
46.7
54.9
72.5
85 .'9
58.5
47.5
14.3
15.4
P04
*T
0.085
—
0.057
0.320
0.290
0.088
0.088
0.180
0.670
0.660
0.04(
0.04*
Suspended Solids
92
42
45
211
221
25
20
27
44
22
22
124
127
46
72
Chloride
9.3
10.2
10.1
7.8
8.2
17.7
14.7
10.1
10.7
14.2
14.2
12.8
13.4
6.4
6.1
Organic N
6.1
<0.05
2.5
4.1
6.2
7.8
7.5
12.2
12.9
Arsenic
2.4
2.1
6.0
3.0
1 .9
r\ r\
Lm * Cm
3.3
1.7
Cadmium
<0.05
0.67
0.10
<0.05
<0.05
<0.05
<0.05
<0.05
Chromium
<0.2
<0.2
2.5
1.4
0.4
0.4
<0.2
<0.2
-------
Parameter*
1
2
3
TABLE 15 (continued)
4 5a
6
6R
LS
Cobalt
<0.5
<0.5
<0.5
j-y mm
<0.5
<0.5
<0.5
<0.5
<0.5
Copper
5.6
2,1
17
53
60
97
25
17
Iron
450
310
570
290
230
430
670
230
Lead
Q 1
0 * 1
3.4
2.4
1.7
1.3
2.2
1.0
<0.1
Manganese
31
24
110
48
CO
Do
170
51
4
Nickel
3.6
<2
<2
2.3
<2
2.3
<2
<2
Selenium
2,1
2,1
1.5
1.5
1.8
2.1
1,3
1.5
Zinc
3.2
6.5
20
5.0
5.2
6.5
2.5
1.4
Inorganic Mercury
<0.05
0.08
<0.05
<0.05
<0.05
0.07
<0.05
<0.05
Organic Mercury
-------
-59-
INTERSTITIAL WATER
The interstitial water was obtained by high speed centrifugation under
a nitrogen atmosphere and stored in teflon bottles for chemical analysis and
bioassay tests. This procedure was described at the beginning of SECTION 7.
Analysis procedures are given in Appendix A and the results in Tables 16 and
17.
Portions of the interstitial water were used in 1977 bioassay tests
(SECTION 8). The water was not filtered in order to preserve its character-
istics as nearly as possible to the water which might be released to the
water column upon dredging or be present in the sediment for exposure to
burrowing aquatic animals. The chemical analyses were performed on the un-
filtered water to characterize the systems used in the bioassay tests. It
is likely that many of the chemical species (such as metals) are associated
with very fine (possibly colloidal) particles in the water (Leckie and James,
1974). In addition, significant portions may be complexed or adsorbed to
organic or inorganic species. Complexation of metals can greatly reduce their
toxicity characteristics as compared to the dissolved aquo forms (Andrew, et
al_. 1977).
The variation in metal content between filtered and unfiltered samples
is shown in Table 17. The arsenic, cadmium, chromium, copper, iron, lead
and zinc concentrations in the filtered samples were generally much less
than found for the unfiltered samples. Only the manganese concentrations
did not change much upon filtering the samples (0.45 micron pore).
For most of the metals investigated, there were no large differences
between interstitial waters of the harbor sediments ana the Lake Superior
sediment (LS). However, unfiltered interstitial water from the Lake Superior
site sediment was generally lower in iron, manganese and zinc than unfiltered
interstitial water from the harbor sediments.
-------
TABLE 16
CHEMISTRY OF
INTERSTITIAL
WATERS -
1977
S
I T E
Parameter*
1
2
3
4
5a
6
6R
LS
Pokegam*
pH
7.1
7.2
6.0
—
?!
6.7
7 1
COD
50.6
52.9
285
99.4
175
171
115
119
77.7
78.1
*146
145
208
224
70.6
71.7
93,6
89.9
H^S
<0.32
<0.20
<0.17
<0.17
0.19
44
0.20
<0.17
<0.17
CI
61.4
62.5
8.9
7.9
21,4
21.9
*1 /") /*
1 L . 0
13.0
13.3
14.6
24,0
00 Q
Co « Q
18.5
18.4
4.18
4.11
12.1
11.9
NH3
2.6
11.7
21
15.5
11
3.8
20
16
5
TKN
3.7
13.3
21
30
22
7.6
46
16.5
17
Total PCBs
0.2
0.5
1.4
3.5
0.4
0.7
___
—
3.1
Organic N
11
1.63
0
14.5
11
3.8
26
0.5
12
Suspended Solids
35
45
40
51
83
66
45
118
41
36
27
Arsenic
5.0
4.1
3.8
3.8
2.8
6.0
5.5
6.1
10
Cadmi urn
<0.05
0.32
2.0
0.05
<0.05
0.30
0.10
2.0
l g
Chromium
0.5
0.5
7.1
4.4
1.1
7.1
3.7
1,5
12
Cobalt
1.3
3.9
<0.5
0.7
<0,5
1.2
<0.5
<0.5
0.7
Copper
11
11
153
24
12
36
15
C(\
OU
10
Iron
13100
570
8100
8000
1700
8200
21400
1400
8100
Lead
0.9
0.9
23
4
3
13
12
6
4
Manganese
6000
6000
960
2450
1400
2300
3200
540
1000
Nickel
<2
<2
9.5
2.3
<2
2.3
<2
7.8
3.6
-------
TABLE 16 (continued)
Parameter 1 2 3 4 5a 6 6R LS Pokegama
Selenium 1 4 12 2 2 1 4 4
Zinc 47 10 68 10 7 14 10 10 10
Total Mercury 0.05 1.7 1.1 0.86 0.32 0.42 0.23 0.44 0.15
*Values listed in mg/1 except for PCBs and metals which are given in yg/1.
-------
TABLE 17
METAL CONTENT OF INTERSTITIAL WATERS* - 1978
SITE
Metal 1 2 3 4 4a 6 LS
U
F
u
F
U
F
U
F
U
F
U
F
U
p
Arsenic
3.3
±0»5
<1.0
7.1
±1.3
<1.0
3.2
±1.5
<1.0
1.3
±1.0
<1 .0
—
—
5.3
db 2 S
3.8
±2.9
6.4
±1.9
4.7
±0.4
Cadmium
0.77
+0 # 5
<0.05
0.47
±0.12
0.09
±0.03
0.63
if).06
<0.05
0.4
±0.07
0.10
±0.10
1.3
±0.2
0.16
±0.04
0.55
±0.37
0.41
±0.05
1.5
±0.1
---
Chromium
2.2
±0.3
<1.0
2.5
±0.1
<1.0
6.7
±0.6
1.7
±0.3
5.1
±1 ,3
<1.0
4.4
±0.7
1 .5
±0.2
6.0
±5.9
1.5
±0.5
<1.0
<1 .0
Cobalt
<1.0
—
<1.0
1.0
±1.0
1.5
±0.2
<1.0
<1.0
<1 .0
<1.0
<1.0
<1.0
<1.0
<1.0
<1 .0
Copper
18.7
±3.4
2.0
±1.4
±12
4.8
±0.5
27.2
+4 7
2,5
±0.5
17.8
±7«7
1.5
±0.4
69
±16
6.7
±0.2
92
±21
56
±29
26.3
±8.9
18.9
±6.5
Iron
5.0
±0.8
0.49
±0.08
17.9
j g
0.35
±0.18
24.7
+2.8
18.2
±3.7
23.5
±4.2
12.7
±4.8
15.2
±5.2
1 .2
10.2
7.4
±1.8
3.2
±0,7
0.32
±0.15
Lead
14 ?
1 » • C>
±3.6
1.5
±1.4
8.2
±5.1
—
22.7
±3.8
1.0
±0.2
10.3
±4 a 4
0.3
±0.2
3 5
il "4*
0,7
±0.7
5.5
"I"12
3.2
±1.0
5.9
±0.8
3.4
Manganese
1.30
±0.03
1.06
±0.02
3.9
±0.5
3.0
±0.6
1,81
±0.03
1.76
±0.16
1.89
±0.41
1.65
±0.37
1.61
±0,02
1J6
±0.04
1.98
±1.02
1.87
±1 .0
1.61
±0.06
1.51
±0.06
Nickel
1.1
±1.1
<1.0
<1.0
<1.0
2.7
±1.1
<1.0
2,4
±0.9
1.4
±1 \ t
—
<1.0
1.3
+*| 4
1.2
i o 7
3.7
±0 „ 5
3.5
+ "| Q
Zinc
42
±44
—
16
±12
13
±12
34
±2
5,4
±2.2
27
±15
4.3
±0.5
31
±1
8.8
±4.9
20
i 6
23
±8
5.2
±0.3
^Concentrations are in yg/1 except for iron and manganese which are listed in mg/1. The uncertainties in the
values are standard deviations based on triplicate analyses at each site.
-------
-63-
Compared to the overlying water, the unfi1tered Interstitial water was
generally greatly enriched in iron (by factors of 15 to 100), manganese (by
factors of 10 to over 100) and mercury (by factors of 5 to 20) even though
the concentrations of suspended solids were about the same. In addition, the
interstitial water generally contained higher amounts of arsenic and about
two to ten times as much chromium and zinc. Even in the filtered interstitial
water samples, iron and manganese concentrations were generally 2 to 100
times greater than in the overlying water.
A comparison of the sites generally does not show large variations in
parameters for the 1977 sampling. However site 3 sediment interstitial water
contained the highest amounts of copper (153 yg/1), lead (23 yg/1), nickel
(9.5 yg/1) and zinc (68 yg/1). Variations between sites 6 and 6R were large
for COD, nitrogen, and some of the metals. This variation may be due, in part,
to the higher level of suspended solids in the intersitial water for sample 6R.
ELUTRIATE WATER
The procedures for evaluating dredged material for potential disposal
effects include an investigation of solid, liquid and particulate phases re-
sulting from the mixing of one volume of sediment with four volumes of water
(EPA/Corps of Engineers, 1977). The water phase obtained after the mixing
process has been called elutriate water (Keeley and Enger, 1974). The elutriate
test has been evaluated for its appropriateness in predicting the pollution
characteristics of dredge material (Lee, et al_., 1975; Cheam, e_t al_., 1976).
In this study, liquid and particulate phase systems were used in certain
of the acute toxicity bioassay tests (SECTION 8). The liquid phase obtained
by mixing test sediments with Lake Superior water was characterized by analysis
of an array of physical and chemical parameters.
The procedure used in preparation of the liquid and particulate phase
systems generally followed the EPA/Corps of Engineers guidelines (1977).
-------
-64-
One variation in proceudre involved elimination of the filtering step for ob-
taining the liquid phase. It was replaced by using high speed centrifugation
to remove about 99$ or more of the suspended material from the particulate
phase. Filtering was omitted since it would be lengthy and risk causing changes
in the liquid phase through such processes as loss of volatiles, oxidation of
components and loss of organics and metals on the filter and retained solids.
A second variation involved using two sediment samples from cores from
each of the 1977 sites in preparing the liquid phase. One sediment sample
was exposed to air and the liquid phase resulting from mixing the sediment
with Lake Superior water is called oxic elutriate water. A second sediment
sample from each site was not exposed to air but the sediment-water suspen-
sion was produced under a nitrogen atmosphere. The liquid phase resulting
from the mixing of the unexposed sediment with Lake Superior water is called
anoxic elutriate water. The Lake Superior water was well-oxygenated in both
cases. Consequently, the terms oxic and anoxic are used here only to designate
whether or not the sediment had been exposed to air.
The two sediment samples were used to investigate possible chemical
releases and potential toxic effects resulting at the dredging site (releases
from sediment not exposed to air) and at the disposal site (sediment exposed
to air).
Preparation of Liquid Phase (Elutriate Water) and Particulate Phase
Sediment from a given sampling site was obtained from the upper portions
of several cores. The sediment was added to a 3 1 beaker containing about
400 ml of Lake Superior water (obtained from the Environmental Research
Laboratory-Duluth, Minnesota) with mixing until the total volume reached
1400 ml. The suspension was transferred to a 12 1 round bottom flask and
enough Lake Superior water was added to produce 5 1 of suspension (1 1 of
sediment - 4 1 of water). The flask was placed on a shaker bath and shaken
for 0.5 hr at 100 excursions per minute. The suspension was allowed to settle
-------
-65-
for 1 hour. (For the 1978 bioassay tests, a portion of the overlying suspen-
sion (particulate phase) was removed). The overlying suspension was trans-
ferred to 250 ml centrifuge bottles (stainless steel for portions to be
analyzed for organics and polycarbonate for portions to be analyzed for
metals) and centrifuged for 15 minutes at 10,000 rpm. The supernatant liquid
(liquid phase elutriate water) was decanted into teflon bottles for use
in subsequent chemical and biological tests. For producing the liquid
phase anoxic elutriate water, the mixing of sediment with Lake Superior
water was carried out in a glove bag under a nitrogen atmosphere.
The elutriate water used in the 1978 bioassay tests (SECTION 8) was
obtained by recentrifuging the supernatant from the first centrifuging at
14,000 rpm for 10 minutes.
Liquid Phase (Elutriate Water) and Particulate Phase Tests
Portions of the liquid phases were used in bioassay tests run in
1977 and 1978. Portions of the particulate phases were used in 1978 bioassay
tests (SECTION 8). The liquid phase (formed by use of sediments from the
various sites) was analyzed for a number of physical and chemical parameters.
General water analysis procedures are given in Appendix A and the analytical
results are listed in Tables 18 through 21.
Survey of Chemical Properties of Elutriate Water
A general survey of Tables 18 and 19 shows that the exposure of
sediments to air prior to use in elutriate water preparation did not greatly
alter the chemical water parameters from values found for sediment unexposed
to air. The elutriate waters produced from air exposed sediments had
higher ammonia values (by about a factor of two) for most of the sites.
However the other chemical parameters do not show any definite trends caused
by exposure to air.
Table 20 indicates that the dissolved oxygen concentrations in the
liquid phase remained above 3 mg/1 in all cases and above 5 mg/1 in most
-------
TABLE 18
CHEMISTRY OF ELUTRIATE WATER (ANOXIC SEDIMENT CONDITIONS) - 1977
SITE
Parameter* 1 2 3 4 5a 6 6R LS Pokegama
pH
6.9
6.9
—
6.0
6.9
5.8
6.4
7.1
—-
H2S
<0.17
0.20
0.20
0.22
<0.17
<0.17
0.20
<0.17
<0.17
Ammonia
5.5
1.8
10.5
13
7
8
12
11
3.0
TKN
5.8
6.8
24.5
19
16
11
26.5
11
5.3
Organic N
0.3
5
14
6
9
3
14
<2
2.3
COD
28.9
241
90.8
101
52.7
111
84.0
41.8
92.1
Total PCBs
0.3
0.2
1.4
2.5
0.9
0.9
—
—
2.6
Arsenic
5.6
7.3
—
6.0
3.4
6.5
6.2
4.3
Cadmium
0.20
0.15
—
0.85
0.05
0.10
0.05
0.20
—
Chromium
8.0
1.4
—
15,0
1.8
3.3
4.1
5.9
---
Cobalt
2.7
0.8
—
<0.5
<0.5
0.5
0.8
0.7
—
Copper
35
9
—
37
17
32
18
21
—
Iron
4400
800
—
1900
2100
2100
3600
2900
Lead
4.1
1.7
—
n
6.7
3.3
6.0
1.7
---
Manganese
285
910
—
345
215
350
350
350
—
Nickel
6,4
<2.0
—
<2.0
<2.0
2.3
<2.0
23
Selenium
1
2
—
4
2
1
1
2
Zinc
21
5.9
—
22
17
18
57
34
Mercury
<0.10
0.25
0.16
0.21
<0.10
0.22
<0.10
<0.10
<0.10
*Va1ues listed in mg/1 except for PCBs and metals which are given in yg/1.
-------
TABLE 19
CHEMISTRY OF ELUTRIATE WATER (OXIC SEDIMENT CONDITIONS) ¦
- 1977
SITE
Parameter*
1
2
3
4
5a
6
6R
LS
Pokegam*
pH
7.0
7.2
—
6.0
6.9
5.6
6.5
7.4
w* W «
H2S
<0.17
0.17
0.20
0.18
0.19
0,44
<0.17
<0.17
—
Ammonia
10
6.7
16
22
11
16.5
20
8
3.5
TKN
10
7.2
24.5
24.5
22
17
38
11.5
7.5
Organic N
<2
<2
8.5
9 C
JLt *
11
<2
18
3.5
4.0
COD
42.7
222
105
82.4
54.3
94.4
116
54.0
88.7
Total PCBs
0.5
0.2
1.2
2.5
0.3
0.9
—
—
1.6
Arsenic
16.5
6.5
—
9.5
5.6
8.4
8.0
7.7
—
Cadmium
—
<0.05
—
0.30
0.05
0.25
0.35
0.20
—
Chromium
9.7
0.3
—
2.5
3.7
7.1
9.0
3.4
—
Cobalt
2.8
<0.5
—
A f
<0.5
a r
<0.o
1.3
0.5
1.6
- —
Copper
10
3
36
11
15
31
21
—
Iron
4700
400
2800
1600
830
5200
2900
2300
—
Lead
2.9
0.4
12
4.0
16
13
1.2
—
Manganese
308
960
—
515
390
525
490
225
—
Nickel
9.3
<2.0
—
2.3
<2.0
2.3
g j
5.0
Selenium
—
2
—
Ij.
2
2
2
1
—
Zi nc
28
1.5
—
22
13
26
58
38
—
Mercury
<0,10
<0.10
0.14
<0,10
<0.10
0.41
0 = 12
<0.10
<0.10
~Values listed in mg/1 except for PCBs and metals which are given as pg/1.
-------
-68-
TABLE 20
PHYSICAL AND CHEMICAL DATA* FOR LIQUID PHASE ELUTRIATE WATER - 1978
Suspended Solids
Specific (Liquid (Particulate
Date
Site
pH
D.O.
COD
JH3-
0.48
Conductance
Phase)
Phase)
June 5
1
6.6
5.4
165
104
105
14,100
June 26
2
6.4
7.6
23
1.3
153
24
5,600
July 15
2
6.5
5.6
31
1.3
110
63
11 ,800
June 19
3
6.6
3.4
3.2
120
101
7,900
June 12
4
6.7
5.6
107
2.7
105
73
9,300
July 15
4a
6.4
5.2
44
1.2
97
119
15,600
June 26
5
6,3
4.9
38
0.5
71
79
10,700
July 10
5
6.3
6.3
43
1.1
71
74
3,200
July 10
6
6.6
4.2
36
0.3
102
141
15,400
June 5
LS
6.6
7.5
109
0.5
100
28
1,900
June 12
LS
6.7
7.8
3
0.5
100
28
4,000
June 19
LS
7.0
8.4
—
0.15
100
60
3,300
June 26
LS
6.2
4.8
14
0.5
68
69
7,600
July 10
LS
6.4
5,3
13
0.2
89
49
5,400
July 17
LS
6.6
7.3
18
2.3
133
25
2,200
July 24
LS
—
23
___
69
6,700
*Units are mg of 02/1 for DO and COD, mg/1 for IMH3, ymhos/cm for specific
conductance and mg/1 for suspended solids. pH, D.O., COD and NH 3 were measured
on liquid phase (centrifuged) elutriate.
-------
TABLE 21
METAL CONCENTRATIONS* IN LIQUID PHASE ELUTRIATE WATER
1978
Date Site As Cd
U F U F
Cr
U F
Co
U F
Cu
U F
Fe
U F
Pb
U F
Mn
U F
Ni
U F
Zn
U F
June 2
1
9.6
2.6
0.64
0.78
7.2
2.2
2,7
<1
0
24
11.4
5.0
1 .15
3.1
1 .0
0.36
o
ro
OO
8.1 3.9
24
9.3
June 26
2
5.0
1.5
0.05
0.06
1 A
1 § «
<1.0
<1.0
<1
0
6.5
3,6
0.60
0.32
1.2
0.4
0.75
0.82
<1.0 <1.0
5.2
6.1
July 15
2
3.7
2.9
0
57
0.40
27
<1.0
1.1
<1
0
42
9.3
2.4
0,32
3.7
0.4
0.50
0.38
3.1 2.3
13
16
June 17
3
8.0
2.1
0
20
0.05
2.6
<1.0
1.1
<1
0
1?
2.6
3.1
0.12
3.7
0.1
0.24
0.18
3.9 <1.0
17
4.2
June 8
4
10.5
3.2
0
20
<0.05
4.5
<1.0
1.1
<1
0
17
2.3
3.7
0.41
4.5
0.2
0.26
0.21
2.0 <1.0
15
4.2
July 15
4.9
1.6
0
23
0.31
5,2
1.2
1 ,3
<1
0
45
7.8
5,5
0.30
10
0.5
0.28
0.13
o
CM
CO
l-v.
28
13
June 26 '
5
9.5
1.0
0
18
0.05
4,6
<1.0
1.2
<1
0
21
1.6
2.8
0.14
d q
nr *
0.3
0.09
0.06
3.0 <1,0
22
5.9
July 6
5
4,9
2i
0
08
0,45
4.2
<1.0
<1.0
<1
0
20
4.1
2.9
0.69
7.2
0.7
0.16
0.10
3.2 <1.0
25
16
July 24
5
3.9
3.3
0
08
0.21
3.3
<1.0
<1.0
<1
0
20
3.2
2.7
0.06
7.0
0.2
0.14
0.07
2.8 <1.0
24
5,6
July 6
6
15
4.9
0
85
- 0.08
9.0
1.1
1 .0
<1
0
35
7.3
5.8
0.74
19
1.3
0.24
0.16
6.2 <1.0
~T75 ;
July 24
6
<1.0
1,4
0
08
0.65
2.6
—
<1 .0
<1
0
19
6.8
2.6
0.74
3.3
0.4
0.32
0.24
3,9 2.0
21
15
June 5
7.0
4.7
0
68
0.30
1.7
<1.0
1,0
<1
0
13
4.9
0.86
0.56
1,6
0.7
0.91
0.77
<1,0 <1.0
8
4.6
June 12
LS
6.2
1.6
0
65
0.26
1.8
<1.0
<1 .0
<1
0
28
7.5
1.5
0.54
1.3
0.3
0.58
0.58
1.1 <1.0
8
4.2
June 19
LS
9.8
4.9
0
97
0.26
2.4
<1.0
1 .0
<1
0
22
9.2
1.5
0,49
2.6
1,2
0,96
0.82
2.0 4.0
9
5.1
July 10
LS
11
6.2
1
2
0.46
2.5
<1.0
1.1
<1
0
17
7.2
2.4
0.74
2.9
0.6
0.50
0.36
3,5 1.1
13
9.5
_ mm 1 r-y,
July 18
LS
9.8
9.2
0
36
0.33
<1.0
1.2
1.0
<1
0
19
11 8
0.74
0.20
3.6
0.1
2.53
2.04
2.0 1.1
7
10
July 24
LS
5.2
2.9
0
88
0.13
1.6
<1 .0
1 .6
<1
0
26
0.8
2.0
0,48
3.3
<0.1
0.75
0.56
4.4 1.8
11
1.6
*Units for metal concentrations are pg/1 except for Mn and Fe which are mg/1. The U and F headings represent
unfiltered and filtered (0.45 micron pore) liquid phase elutriate water.
-------
-70-
cases. The particulate phase contained about two to fifteen grams per liter
of suspended material.
Table 21 indicates the effect of filtering the liquid phase on metal con-
centrations. Metals concentrations were generally decreased upon filtering.
This could result from either adsorption on the filter paper (cellulose acetate)
or removal of particulates along with their adsorbed metals. Considering iron
and manganese, there was a much greater per cent decrease in the concentrations
of iron than manganese. This greater effect of filtration on iron concentrations
could indicate the presence of particulate iron in the form of oxides or hydrous
oxides in the liquid phase elutriate water which are largely removed from solution
upon filtration.
Comparison of the chemical parameters of the Lake Superior site to those
for the harbor sites does not generally show large differences. Except for
one instance, the LS liquid phase elutriates had lower COD and NH3 values. The
metal concentrations were similar in the elutriate waters (filtered and unfiltered)
for the LS samples as compared to harbor site samples. The reason for one
particularly high manganese concentration in the LS elutriate water from July
18, 1978 sampling is unknown.
GENERATED PORE WATER
Certain fractions of sediment chemical consituents can potentially be re-
leased to the water. This release may be a diffusion process influenced by
diagenesis of the minerals in the sediments or it could occur upon disturbing
the sediments and mixing them with the overlying water. The released chemical
species may be dissolved in the water or associated with fine particulate material
which can remain suspended for long periods.
In conduction with assessing potential harmful effects of dredged sediments,
some experiments were carried out to provide information on the rate and amount
of release of chemical species from sediments when mixed with "pure" water.
-------
-71-
After removal of interstitial water from a given sediment sample, the same volume
of Lake Superior water was mixed with the sediment, allowed to remain in contact
for a certain period (described below), and removed. This sediment extraction
process with Lake Superior water was repeated several times without directly ex-
posing the sediment to air. The Lake Superior water extracts were analyzed for
COD, D.O., pH, certain metals and, in some cases, PCBs. From the concentrations
of these chemical species in the Lake Superior water extracts and the weights of
sediment used, the amounts of chemical species released per kg of sediment were
computed. The Lake Superior water extracts are called generated pore water.
Generated Pore Water Production Procedure
Sediment was added to ten weighed stainless steel centrifuge bottles under
a nitrogen atmosphere and reweighed. The bottles were centrifuged at 10,000
rpm for 15 minutes using a refrigerated centrifuge (4°C). The centrifuge
bottles were placed in a glove bag under a nitrogen atmosphere, the caps were
removed and the interstitial water was decanted into teflon bottles and its
volume recorded. The caps were replaced and the bottles and sediment weighed.
The interstitial water was recentrifuged in polycarbonate centrifuge bottles
at 14,000 rpm for 30 minutes and the supernatant was collected for chemical
analyses.
The sediment was removed from the stainless steel centrifuge bottles and
placed in a blender under nitrogen. A small portion was used to determine the
percentage of water in the sediment. A volume of Lake Superior water (equal to
the volume of removed interstitial water) was added. The sediment and water
were blended at low speed for five minutes and the resulting slurry was placed
back into the ten centrifuge bottles and capped. The bottles were refrigerated
until the water was to be removed by centrifugation and Lake Superior water
again added using the same procedure. The centrifuge bottles and sediment
were weighed each time after the removal of the water. After the initial
-------
-72-
removal of interstitial water and addition of Lake Superior water to the
sediment, the process was repeated after 12 hrs, 1.5 days, 3.5 days, 7.5
days, 10.5 days, 15.5 days and 21.5 days.
During one experiment, two stainless steel centrifuge bottles filled
with Lake Superior water were carried through the same water removal and
water addition process. The water was analyzed for metals to test for
leaching from the stainless steel containers.
The procedure described above was completed for sediment from site 4R.
In addition, some data was obtained using site 6R sediment but a breakdown
in the centrifuge prohibited obtaining data after 10.5 days. An initial
experiment was carried out using site 6 sediment but weights were not re-
corded and only chemical specie concentration- values were obtained for
the generated pore water.
Some data was also obtained using polycarbonate bottles in place of
stainless steel. The effect of blending the sample on concentrations of
chemical species in the water was investigated.
Generated Pore Water Results
The results of the initial study of the concentrations of chemical
species in the initial interstitial water and generated pore water for
site 6 sediment are given in Table 22. Also included is data on the values
of these chemical species in Lake Superior water and in Lake Superior water
kept in the stainless steel centrifuge bottles (blanks). The values of the
chemical parameters in the Lake Superior water and blanks are all low com-
pared to the water obtained by using the site 6 sediment. For the generated
pore water, the values of the chemical species tend to increase over those
found for the interstitial water (initial values) except for manganese.
Significant amounts of the chemical species continue to be extracted even
after seven extractions (21.5 day values). These water samples were not
filtered and thus the chemical values may include contributions from species
-------
TABLE 22.
INTERSTITIAL AND GENERATED
PORE WATER FOR
SITE 6*
.
Time
As
Cd
Cr
Cu
Fe
Site 6 Pore Water
Pb Hn
Ni
Zn
Inorganic Total
Hg Hg
Co
COD
Initial
5.1
0.05
4.6
18.0
9.6
8.7
2400
<2.0
14
0.17
—
<0.5
146.2
12 hr.
15.8
21.5
1.10
1.45
15.2
18.8
105
77
7.1
7.1
65
1400
1100
14.1
17.1
56
0.24
0.22
0,8
145.0
635.9
1% day
29.4
0.95
19.0
97
10.4
112
1890
10.8
79.2
0.54
0.51
0.6
893.2
3% day
10.7
1 .40
18.9
130
10.6
94
1280
19.3
73,6
0.3
0.2
0.8
834.3
lh day
10.7
1.05
5.4
28.5
3.8
9.8
1940
6.9
26
0.13
0.38
<0.5
283.9
10% day
21.2
18.95
8.9
54
3.9
17.1
1840
31.7
80
3.08
3.09
<0.5
323.6
15% day
8.9
0.40
7.0
36.4
3.5
12.2
1670
10.1
62
0.05
0.42
<0.5
196.1
21*2 day
7.2
0.20
7.3
44
2.4
Lake
6.0
Superior
1890
Blanks**
6.9
25.6
0.26
0.23
<0.5
167.2
12 hr
<0.5
<0.05
1.1
4.9
0.018
11.9
2.1
2.2
<0.5
1% day
0,9
<0.05
<0.2
9.5
0.038
0.2
5.3
<2.0
2.3
<0.5
lh day
0.6
<0.05
<0.2
32.7
0.012
0.2
17.8
2.3
3.9
<0.5
10% day
0.7
<0.05
!<
6.6
0.011
0.2
4.6
<2.0
6.3
0.14
0.55
<0.5
Lake Superior
Water 1.1 <0.05 <0.2 0.9 0.013 0.4 1.9 <2.0 2.4 <0.5
* All values are in yg/1 except for Fe and COD which are in mg/1. Se was less than one yg/1 in all samples
**Blanks were in stainless steel cups.
-------
-74-
associated with fine suspended particulate material not removed during
centrifuging.
Experiments with sites 4R and 6R sediments included measurements of
the weights of sediments used and this allowed calculation of the amount
of chemical releases per kg of dry sediment. Table 23 includes data on
sediment weights, volume of water removed from the sediments, pH and dis-
solved oxygen values. The pH values are seen to remain in the 6.2 to 6.9
range for the extracted water. The dissolved oxygen levels in the water
drop as extractions proceed but eventually increase for the latter extrac-
tions. The pH of Lake Superior water used in producing the generated pore
water was about 7.5 and the dissolved oxygen levels were about 7 to 9 mg/1.
The concentrations of chemical parameters for the interstitial and
generated pore water for sites 4R and 6R are given in Tables 24 and 25.
Most of the chemical parameters were higher in the generated pore water
samples than found for the initial interstitial waters (exceptions are
iron and manganese). The values for COD, iron, chromium and lead tend to
decrease during the latter extractions. The data for site 4R covers seven
sediment extractions while that for site 6R includes five extractions.
Tables 26 and 27 list the total number of mg or ug of the chemical
parameters removed from the sediment during the water extraction process.
Considering the values for the metals, iron and manganese gave the largest
extracted amounts corresponding to the much higher abundances of these
metals in the sediments. The amount of release per kg of sediment follows
the trend of concentrations in the water extractants. Comparing the two
sites, the amounts of extracted chemical parameters are similar.
Discussion of Results
The application of the generated pore water results to the environmental
effects of sediment dredging is difficult. However the data may give a
-------
TABLE 23
GENERATED PORE WATER DATA
-75-
S i t e 4R
Time Wet Weight Dry Weight Volume 1^0 pH D.O.
Initial
1978.53
g
1163.38
g
817
ml
6.40
3.3 ppm
12 hr.
2078.99
g
1222.45
g
518
ml
6.39
1.35 ppm
1H day
1768.75
g
1040.03
g
450
ml
6.30
0.65 ppm
3h day
1515.24
g
890.96
g
390
ml
6.25
0.35 ppm
7h day
1173.63
g
690.09
g
455
ml
6.20
1.20 ppm
10^ day
1012.10
g
595.12
g
430
ml
6.85
2.35 ppm
15*5 day
926.87
g
544.99
g
500
ml
6.69
2.60 ppm
21^ day
822.30
g
483.51
g
440
ml
6.60
5.10 ppm
S i •
t e
6R
Time
Wet Weight
Dry Weight
Volume h^O
pH
D .0.
Initial 2631.44 g 1539.39 g 765ml 6.70 3.10 ppm
12 hr. 1959.77 g 1146.46 g 590 ml 6.70 2.10 ppm
1% day 1921.86 g 1124.28 g 565 ml 6.71 0.60 ppm
3h day 1493.69 g 873.81 g 670 ml 6.70 0.40 ppm
7^ day 1378.40 g 806.36 g 720ml 6.58 1.90 ppm
103s day 1312.23 g 767.65 g 705 ml 6.62 5.10 ppm
-------
TABLE 24
CHEMICAL PARAMETERS* IN INTERSTITIAL AND GENERATED PORE WATER FOR SITE 4R
COD Fe Mn Inorganic Total
Time (mg/1)(mg/1)(mg/1) Ni Pb Cu Cr Zn Cd Hg Hg
Initial
138
12
2.3
<2
7
23
6
13
<0.5
<0.05
0.2
11
12 hr.
797
10
1.1
12
150
96
7
91
1.7
1.5
1 .9
28
1.5 day
667
7
1.0
14
75
123
21
56
2.4
2.7
3.3
14
3.5 day
>1000
11
1.4
19
179
166
17
91
2.0
1.2
1 .4
54
7.5 day
423
5
1.0
20
37
94
5
80
1.8
0.2
0.2
17
10,5 day
318
5
1.3
20
35
98
10
94
1.2
10
15.5 day
130
3
2.1
23
8
56
4
66
1.9
0.3
0.9
14
21.5 day
117
2
2.2
25
2
126
7
130
0.8
0.2
0.2
6
*Values are in pg/1 unless designated otherwise.
-------
-77-
TABLE 25
CHEMICAL PARAMETERS* IN INTERSTITIAL AND GENERATED PORE WATER FOR SITE 6R
COD Fe Mn Inorganic Total
Time (mg/1) (mg/1)(mg/1) Ni Pb Co Cu Cr Zn Cd Hg Hg
Initial
435
28.0
3
58
3.0
6.8
<0.5
17
6.7
56
<0.05
0.22
0.35
12 hr.
1508
12.7
1
94
33.7
119
0.9
173
40.7
156
3.0
1.42
1.82
1.5 day
1316
12.9
1
75
18.9
137
0.8
160
46.0
62
1 .8
1.06
1.13
3.5 day
608
2.3
0
87
17.5
2.0
0.6
83
7.1
96
0.95
1.59
1.43
7.5 day
232
5.8
1
82
30.0
24
<0.5
37
8.5
88
1.05
7.5
8.89
10.5 day
163
5.3
1
66
25.0
24
0.6
71
11.0
64
1 .2
0.27
0.32
Concentrations are in yg/1 unless designated otherwise.
-------
TABLE 26
CHEMICAL RELEASES FROM SITE 4R SEDIMENT
COD
Fe
Mn
Ni
Pb
Cu
Time
mg
released mg*/kg
mg
released
mg/kg
mg
released
mg/kg
ug
released
va./,k9.
pg
released
ug/kg
ug
released
ug/kg
Initial
113 97
9.8
8.4
1.9
1.6
<
<1.4
5.7
4.9
19
16
12 hr.
413 334
5.2
4.2
0.6
0.5
6,2
5.1
82
67
50
41
1.5 day
300 289
3.1
3.0
0.4
0.4
6.3
6.1
34
32
55
53
3.5 day
>390 >438
4.3
4.8
0.5
0.6
7.4
8.3
70
78
65
73
7.5 day
193 279
2.3
3.3
0.5
0.7
9.1
13.2
17
25
43
62
10.5 day
137 230
2.2
3.6
0.6
0.9
8.6
14.4
15
26
42
71
15.5 day
65 119
1.5
2.7
1.0
1.9
11.5
21.1
4
7
28
51
21.5 day
52 107
0.9
1.8
1.0
2.0
11 .0
22.8
1
2
55
115
Total 1900
Total
32
Total
8.6
Total
91
Total
242
Total
480
Cr
Zn
Cd
Inorganic Hg
"J" 4- HK 1
Total
Hg
PCB
Time
ug
released yg/kq
ug
released
ug/kg
ug
released
ug/kg
yg
released
ug
released
ug/kg
u9
released
ug/kg
Initial
5 4
11
9
<0.4
<0.4
<0.04
<0.04
0.14
0.12
9
8
12 hr.
4 3
47
39
0.9
0.7
0.78
0,64
0.98
0.81
14
12
1.5 day
9 9
25
24
1.1
1.0
1.22
1.17
1.49
1.43
7
6
3.5 day
7 7
35
40
0.8
0.9
0*47
0 ^3
0.55
0.61
21
24
7.5 day
2 3
36
53
0.8
1.2
0.09
0.13
0.09
0.13
8
11
10.5 day
4 7
40
68
0.5
0.9
—
—
—
—
4
7
15.5 day
2 4
33
61
0.9
1.7
0.16
0.28
0.45
0.83
7
13
21.5 day
3 6
57
61
0.3
0*7
0.08
0.16
0.08
0.16
3
5
Total 43
Total
350
Total
7.1
Total
3
Total
4
Total
86
^Release per kg of dry sediment.
-------
CHEMICAL
RELEASES
FROM SITE
: 6R SEDIMENT
COD
Fa
r c
Mn
Ni
Pb
Co
Time
mg
released mg/kg*
mg
released
mg/kg
mg
released
mg/kg
yg
released
1 yg/kg
y9
released
yg/kg
y9
released
yg/kg
Initial
333
215
21.4
13.9
2.74
1.78
2.3
1.5
5.2
3.4
< . ¦3
<0.25
12 hr.
889
776
7.5
6.5
1.15
1.00
1Q Q
| J , J
17.3
70.2
61
0.53
0.5
1.5 day
743
661
7.3
c k
• Vr
0.99
0.88
10.7
9.5
77.1
69
0.45
0.4
3.5 day
407
466
1.5
1.8
0.58
0.67
11.7
13.4
1.3
1.5
0.40
0.5
7.5 day
167
207
4.2
5.2
1.31
1.63
21 .6
26.8
17.2
21.4
<0.36
<0.5
10.5 day
115
150
3.8
4.9
1 .17
1.52
17.6
23.0
16.8
21.9
0.42
0.5
Total
2480
Total
38.8
Total
7.48
Total
91 .5
T" o ta 1
178
Total
2
Cu
Cr
Zn
Cd
Inorganic
Hg
Total
Hg
Time
yg
released yg/kg
yg
released
yg/kg
yg
released
yg/kg
released yg/kg
yg
released
yg/kg
yg
released
yg/kg
Initial
13
8,2
5.1
3.3
43
28
<0.04
<0.03
0.17
0.11
0..27
0.18
12 hr.
102
89
24.0
2!
92
61
1.8
1.50
0.84
0.73
1.07
0.93
1.5 day
90
80
26,0
23
35
31
1.0
0 o90
0.60
A CO
9 Kt SJf
0.64
0.57
3.5 day
55
63
4.8
5.5
64
74
0.64
0.73
1.07
1 * 22
0.96
1 .10
7.5 day
27
33
6.1
7.6
63
79
0.76
0.94
5.37
6.7
6.4
7.94
10.5 day
50
OD
7.8
10.1
45
59
0.85
1.10
0.19
0,24
0.23
0.30
Total
338
Total
70.5
Total
332
Ta+ a 1
I Ola 1
5.2
Tnf a 1
I vi w 1
9 5
Total
11 .0
^Release per kg of dry sediment
I <-» J
UD
1
-------
-80-
rough measure of the amount of metals, COD material and PCBs which could
be readily flushed from the sediments upon disposal in a clean water system.
Since the sediments and added Lake Superior water were intimately mixed
by blending, the amounts of chemicals in the water system are probably
upper limits to the amount that would be found in pore water moving through
the sediments. The results for sites 6 and 4R showed that many of the
metal concentrations remained high in the water extractant even after seven
extractions. Consequently, significantly greater amounts of these metals
(particularly manganese, nickel, copper, zinc and mercury) could have
been removed from the sediment with more extractions.
Some additional work was performed using polycarbonate centrifuge
bottles to hold the sediment in place of stainless steel. The change
in centrifuge bottle material had little effect on the COD, arsenic,
cadmium, chromium, cobalt, copper, lead, mangenese, nickel and zinc con-
centrations in the water extractant. Some enrichment of iron in the
water was indicated using stainless steel but this enrichment was not large
compared to the magnitude of the iron concentrations.
The times chosen for allowing the Lake Superior water to remain in
contact with the sediments were arbitrarily picked. Some limited measure-
ments indicate that the concentrations of the chemical parameters in the
pore waters were attained within a few hours of mixing Lake Superior
water with the sediment. One experiment on the effect of blending showed
that this process greatly increased the concentrations of chemical para-
meters (COD and metals).
Using the total release values per kg of sediment for the parameters
given in Tables 26 and 27 and the total concentrations in the sediments
from Tables 2, 9 and 11, the percentage of each parameter extracted by
Lake Superior water can be computed. These values are summarized in
Table 28. The results show that one per cent or less of the most of the
-------
-81-
TABLE 28
TOTAL AND WATER EXTRACTED CHEMICAL PARAMETERS IN SEDIMENTS*
Site 4R COD Fe Mn Ni Pb Cu Zn Total Hg PCB
Total in 137,000 46,900 685 57 94 40 315 0.77 2
Sediment
Extracted 1,900 32 8'6 °-091 °*242 °'480 °'350 °-004 °'085
% Extracted 1.4 0.07 1.3 0.16 0.26 1.2 0.11 0.5 4.3
Site 6R
Sediment 168,000 42,400 545 125 84 19 218 0.23
Extracted 2'480 38,8 7,5 °-092 °-178 0.338 0.332 0.011 —
% Extracted 1.5 0.09 1.4 0.07 0.21 1.8 0.15 5.8
*A11 values of the parameters are in mg/kg.
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chemical parameters in the sediments were removed by the water extractions.
Only the PCB value for sediment 4R (4.3% extracted) and the mercury value
for sediment 6R (5.8% extracted) exceeded the two per cent value. These
results indicate that these substances are not readily available to water
flushing the sediments unless perhaps substantial amounts of particulate
matter is suspended in the water.
-------
-83
SECTION 8
BIOASSAY TESTS
GENERAL BIOASSAY PROCEDURES
Light arid Temperature Control
All bioassays were conducted in an environmentally controlled area of
the University of Wisconsin-Superior wet laboratory. The area was lighted
by an equal number of GroLux Wide Spectrum and Dura-Test Vitilight fluorescent
tubes on a 16 hour photoperiod. Light intensity over the bioassay test area
averaged 85 ftc. Temperature during the bioassays was maintained between
18-20°C by the facilitie's air conditioning system. Tests were conducted
in water baths with temperatures of 17.5-18.5°C. Bath temperature was
maintained by control units designed by Mr. Walter Dawson, US EPA Environmental
Research Laboratory, Duluth.
Daphnia Culturinq Techniques
The parent Daphnia magna stock was obtained from a clone maintained
by the US EPA Environmental Research Laboratory-Duluth. Daphnia were cul-
tured in Lake Superior water in 4 liter glass containers held at 18°C in
the water baths following published procedures (Biesinger and Christensen,
1972), Approximately 5-10 daphnids were transferred using a 10 mm bore glass
tube to the initial 10 culture jars in which 3 liters of Lake Superior water
had been allowed to equilibrate to laboratory temperature and pressure for
12-24 hr. Cultures were fed a concentrated mixture of Cerophyll and en-
riched trout fry granules (Glencoe Mills) blended together with Lake Superior
water, at the rate of 1 ml/liter, twice weekly (Biesinger and Christensen,
1972). Lake Superior water used in both culturing and testing was obtained
from the EPA Environmental Research Laboratory-Duluth. Daphnia from mature
cultures were transferred to additional culture jars to increase the number
of animals for testing. Existing cultures were thinned and transferred to
-------
-84-
new containers containing clean Lake Superior water each week to prevent
overcrowding, male production, crashes and bacteria or algae buildup. On
the day prior to starting each bioassay, all young daphnids were removed
from the stock cultures. This removal insured that young produced during
the 24 hr. period proceeding the test could be identified and removed from
the stock cultures during the day the tests were initiated. The 20-30 stock
cultures generally produced adequate numbers of young daphnids to fill the
requirements for each test. However, in a few instances, some two day old
animals were mixed in with young of the day. Young daphnids were randomly
selected for distribution among the test chambers.
Acclimation and Handling of Test Organisms
Mayfly nymphs, bluegi11s and Pontoporeia affinis collected in the field
(SECTION 6) were acclimated to laboratory conditions from 4 days to six weeks
prior to being used in bioassays. All animals were acclimated in flow-
through aquaria systems. The holding systems were supplied with temperature
controlled dechlorinated city of Superior water. The city's water supply
is derived from a series of shallow horizontal wells extending out under
the bed of Lake Superior from Minnesota Point, and is generally similar in
chemical makeup to Lake Superior water (Appendix C).
Mayfly nymphs were acclimated in a 181 x 35 x 40 cm stainless steel
chamber located in the area where the bioassays were conducted. The bottom
of the chamber was covered with approximately 12 cm of sediment extracted
from the area where the nymphs were collected, prior to placing the nymphs
and water from the same area, into the chamber. Flow of laboratory water into
the chamber was adjusted so that water temperatures were not altered by more
than 1-2°C daily until a temperature of 18°C was attained. Once the acclima-
tion temperature of 18°C was reached, temperature control units were used to
control temperature and flow in the system. The control units responded to any
increase in temperature by opening a solenoid valve resulting in flow of
*
V
-------
cooler water until the temperature declined to 18°C and the solenoid closed.
Oxygen in the chamber was maintained near saturation by air stones placed
at both ends.
Mayfly nymphs were removed from the acclimation chamber on the day
prior to initiation of each bioassay. The removal process involved gently
washing the nymphs from the sediments by placing sediment from the acclimation
chamber on screens which were gently agitated in 18°C laboratory water.
The agitation served to wash away and float the nymphs free of the sediment.
The nymphs were floated off the screens and transferred into a 120 x 18 x
20 cm glass aquaria until the bioassays were started. Temperature, water flow
and oxygen in the glass chamber were maintained in the same manner as de-
scribed for the larger stainless steel acclimation chamber. During the
process of removal, nymphs in their last instar, which was distinguished
by the presence of large dark wing pads, were discarded. This eliminated
the possibility of animal loss through hatching during the bioassays.
Bluegills were acclimated using a system identical to that used for
acclimation of mayfly nymphs. However, the chamber was located in a lab-
oratory area where human activity, lighting and room temperature were sub-
ject to less control. During the acclimation period, lighting was provided
through the laboratory window. Bluegills were treated with a mixture of
Malachite green and formalin to control disease subsequent to arrival in
the laboratory. A concentrated solution of formalin and Malachite green
was added at the rate of 1 ml/liter following the recommendations of Schachte
(1974). Fish were maintained in the holding system for a minimum of one week
following treatment, prior to being used in the bioassays. During the ac-
climation period, bluegills were fed commercial trout pellets on alternate
days.
Bluegills were removed from the holding system three days prior to
-------
-86-
iinitiating the bioassay and placed in the laboratory area test chambers
used during the tests to promote acclimation to the test system.
Pontoporeia collected from Lake Superior (SECTION 6) were acclimated
in a fiberglass chamber measuring 150 x 27 x 35 cm. Pontoporeia were added
to the holding chamber on top of a 0.5 cm layer of Lake Superior sediment.
Temperature was increased from 12°C to 18°C over a two day period and was
maintained at 18°C in the manner described for Hexagenia. Oxygen was main-
tained near saturation by air stones and flow of laboratory water through
the system during the 1-2 week acclimation period. Pontoporeia used in the
tests were collected from the holding chamber by gently stirring the bottom
sediments and passing a fine mesh net through the turbid water.
OVERSEDIMENT BIOASSAY
Methods
Oversediment bioassays were conducted with sediment from all test
sites collected during 1977-1978. Hexagenia limbata, Daphnia magna and
Pontoporeia affinis were used in the tests. The basic tests were designed
to incorporate the mechanisms for transfer of toxic substances between benthic
and plankton communities. In these bioassays, Hexagenia served as a sediment
toxicity probe, sediment toxicant transport mechanism and a means of measur-
ing availability and bioaccumulation of toxicants. Daphnia served as an
indicator of toxicity of materials released from the sediments as a result
of chemical transfer and activity of Hexagenia. Pontoporeia bioassays were
conducted to measure acute toxicity of liquid ohase elutriate water. In
these tests, the particulates removed by centrifuging the suspended phase
were used to form a sediment base (substrate) for the animals, resulting
in the designation as oversediment bioassays.
Hexagenia-Daphnia Bioassay Methods--
Most Hexagenia and Daphnia bioassays were conducted in eight 40 x 20 x
20 cm glass chambers partitioned into two 20 x 20 x 20 cm sections by a
-------
-87-
removeable divider (Figure 4), One side of each chamber was filled in the
field with approximately 8 cm of sediment collected by ponar dredge from the
test site. The filled chambers were covered to protect the sediment from
light and to control temperature fluctuations during the return trip to the
laboratory where an equivalent amount of sediment from the control site was
added on the opposite side of the divider. Sediments from the Pokegama control
site were normally sampled on the same day or on the previous day with a
ponar dredge and held in the laboratory in polyethylene bags at 18°C. In
loading the chambers, only sediments which had not been in contact with the
dredge were employed. When Lake Superior sediments were used as the control
(1978 bioassays), both the control and test site sediments were collected
several days to a week prior to use in the tests. The sediments were stored
in polyethylene containers at 4°C in the dark until placed in the bioassay
tanks. The filled chambers were placed in a water bath at 18°C and covered
for approximately 12-15 hours prior to testing.
The first five tests were designed to determine whether Hexagenia bur-
rowing behavior could serve as an indicator of sediment quality. During
the tests, 4-8 cm of Lake Superior water was added to both sides of the
chambers and the divider was removed. Hexagenia were added, five over the
test sediment and five over the control sediment, one at a time and the
point of burrowing was recorded. After the initial ten insects were added
the divider was inserted, the overlying water was changed and additional
Hexagenia were added so that each side should have contained an equal number.
Because the insects did not inspect the bottom prior to burrowing, burrowed
withing a few seconds of reaching the bottom, and were injured on occasion
by placement of the divider, subsequent tests were conducted with the divider
in position and placing an equal number of Hexagenia on each side of the test
chamber. Most tests were conducted with 10 animals per chamber section.
However, numbers varied between 5 and 12 according to availability of test
-------
-------
FIGURE: 4
SEDIMENT BIQASSAY CHAMBER
Air lines
Chamber divider
i\ ¦% pi Ar%j^iin4iiii nit, *
V/
V/
Modified
beaker
Control Site Sediment
Harbor Site Sediment
-------
-89-
animals in the laboratory. During all tests, individuals which did not
actively swim and burrow within three minutes of being placed in the chamber
were removed and replaced.
After placing the insects in the chambers, two 250 ml beakers were
suspended in the water over the sediments on both sides of each chamber
(Figure 4). Each beaker contained two 25 mm holes covered by 50 y stain-
less screen attached on the inside with sil icone sealant. The modified
beakers allowed for water exchange but prohibited escape of young daphnids.
Five young of the day Daphnia were placed in each beaker at the start of
each test.
Oxygen concentration in the test chambers was maintained and controlled
so that half the replicates (4) were near saturation (6-9 ppm) and the other
half were kept between 1-5 ppm during the 1977 bioassays. Oxygen was main-
tained near saturation in all chambers during 1978. Oxygen levels were
adjusted by controlling air flow through glass pipettes placed in each
chamber (Figure 4). Oxygen concentration and temperature were measured four
times each day throughout the 16 hour photoperiod with a Yellow Springs
Instrument Model 54A system, to identify changes ana minimize variation.
Oxygen flow was controlled by standard aquarium air valves. Water turbidity
in the test chambers was increased by the activity of Hexagenia and was
measured each day. Turbidity measurements were made with a Nephelometer
(Ecologic Instrument Co.) in Formazin Turbidity Units (FTUs). Oxygen,
temperature and turbidity means and standard deviations during the tests
are presented in Appendix C.
Daphnia survivals were determined after 24, 48, 72 and 96 hours. Test
beakers were slowly lifted from the stainless steel wire frame used to
suspend them and most of the water was allowed to flow out the screened open-
ings without impinging the Daphnia. Daphnia in the 3-4 cm deep layer of water
remaining in the beakers were counted over a light box. Animals which did
-------
-90-
not move during counting or respond to being picked up by the glass transfer
tube were considered dead.
Deacl Hexaqenia located at the surface were recorded when turbidity levels
permitted observation. Total counts of surviving Hexagenia were made at
the end of the 96 hr. tests at the time of sediment removal from each side
of the partition. The animals and sediment were separated by the sieving
procedure described previously. Living Hexagenia removed from each chamber
section were rinsed, wet weighed, wrapped in aluminum foil, appropriately
labeled and frozen for subsequent selected organic compound and metal analysis.
At,the end of the 96 hour bioassay tests, samples of the overlying
water were removed and chemically analyzed for pH, total phenols, H^S, NH^, .
TKN, organic N, COD and a number of metals (Appendix C).
Pontoporeia affinis Bioassay Methods —
Bioassays with Pontoporeia were conducted during 1978 in 250 ml poly-
carbonate centrifuge bottles. During these tests, 200 ml of suspended par-
ticulate phase (SECTION 7) was placed in the bottle and centrifuged for
15 minutes at 10,000 rpm. After centrifuging, the appropriate volumes of
the liquid phase were removed and replaced with Lake Superior water to form
5, 50 and 1001 concentrations of liquid phase-Lake Superior water mixtures.
Sediment located on the sides of the tubes was scraped loose and settled to
the bottom to provide a sediment substrate for animals used in the test.
Tests were designed to compare survival in the three aqueous phase - substrate
systems from each test site with survival in identical systems prepared from
the Lake Superior control site sediments and in Lake Superior water (without
a sediment base). A total of 10 bioassays were performed with sediments
from 7 sites. Each test was performed with 3 to 4 replications. In performing
these tests, the test and control site centrifuge bottles were randomly
assigned space in the 18°C water bath and held until temperature equilibrium
occurred. Oxygen concentrations were brought to near saturation and main-
-------
tained by air tubes (glass pipettes). After allowing for appropriate tem-
perature - oxygen adjustments, 6 Pontoporeia were randomly selected from
the laboratory stock and pipetted into each chamber.
Water temperature and the number of animals observed on the water surface
were recorded once daily during the 96 hr tests. Survival was determined from
counts of animals sieved from sediment at the end of each test.
Results
Hexagenia Bioassays —
Survival of Hexagenia in sediments ranged from 97.5 to 33% (Table 29).
Survival was lowest in both the harbor test site 1 sediment and the Pokegama
Slough control site during the bioassay conducted from June 13-17, 1977
(Table 29). Because of limited number of test animals, the test was conducted
with 4 rather than 8 replicates (2 per oxygen level). The limited number of
replicates or a reduction in strength of the test animals apparently resulted
in abnormally low survival. Because of the low survival of control organisms,
data from the test were omitted from all statistical analysis.
Survival of Hexagenia did not fall below 11% in any other bioassay con-
ducted during 1977 with Rainy Lake Hexagenia. Analysis of variance (Steele
and Torrie, 1960) was used to determine whether Hexagenia survival was in-
fluenced by oxygen concentrations during the 1977 bioassays (Blocks) or the
sediment in the chambers (Treatments). The analysis suggested that neither
factor had a significant influence on survival. Because oxygen concentration
did not influence survival, results for replicates subject to varying oxygen
conditions were combined in comparing survival between sediments from
Pokegama Slough, Lake Superior and various harbor test sites (Table 30). Mean
survival was compared by Student's t statistical parameters. Comparison of
means for the 1977 bioassays suggested survival was significantly higher in
test site 3 sediment than in sediment from Pokegama Slough. The 1977 tests
also suggested that survival in site 4 sediment was lower than in Pokegama
-------
TABLE 29
SURVIVAL OF HEXAGENIA LIMBATA IN HARBOR SEDIMENTS AT TWO DISSOLVED OXYGEN LEVELS.
SURVIVAL %
Number
of
Reps.
Animals
per
Rep.
L
o w
°2
High
°2
Value
of F
Level
or
Sign
Date
Site
Control
Test
Control
Test
June 6 - June 10
2
4
9-19
85
5
92.5
92.2
78.0
2.92
NS
June 13 - June 17
1
2
9
61
0
33.0
62.0
72.0
1.83
NS
June 27 - July 1
6
4
9-11
80
5
85.0
82.5
87.5
0.15
NS
July 11 - July 15
3
4
5-11
85
0
93.0
85.0
92.0
0,83
NS
July 18 - July 22
4
3 or 4
5-10
90
0
77,0
90.0
87.0
0.60
NS
July 25 - July 29
5a
4
10-12
84
0
Q? K
ZrC~ «
85.2
87,5
0.33
NS
Aug. 8 - Aug. 12
LS
4
10
97
5
90,0
92.5
95.0
2.15
NS
Aug. 15 - Aug. 19
6R
4
10
85
0
92.5
85.0
oo „u
0.67
NS
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-93-
TABLE 30
SURVIVAL
OF HEXAGENIA LIMBATA
IN HARBOR SEDIMENTS 1977-1978
Date
Site
Reps./Test
No/Rep.
Survival %
Val ue
of t
Control*
Test
1977
June 6-June 10
2
8
9-19
88.9
85.3
0.8
June 27-July 1
6
8
9-11
81.5
86.2
0.7
July 11-July 15
3
8
5-11
85.1
92.8
2.4**
July 18-July 22
4
6-8
5-10
89.9
81.7 .
2.0
July 25-July 29
5a
8
10-12
84.5
90.0
1.0
Aug 8-Aug 12
LS
8
10
95.0
92.5
1.0
Aug 15-Aug 19
6R
8
10
85.0
88.8
0.7
1978
June 12-June 16
4
8
10
87.5
70.0
4.6+
June 19-June 23
3
8
10
71.6
80.7
1.6
* Control represents Pokegama Bay sediments during 1977 and Lake Superior
sediments during 1978.
**Values declared significant with P<0.050
+ Values declared significant with P<0.010
-------
-94-
Slough sediments (P>0.1, Table 30). To establish the validity of the re-
sults, bioassays using sediments from test site 3 and 4 were conducted during
1978, During the 1978 tests, Hexagenia from Ox Creek and Lake Superior sediment
(control) were used. The results were similar to those in Lake Superior sedi-
ment. Survival in sediments from site 3 appeared to be higher than in Lake
Superior sediments.
Observation of Hexagenia indicated that the burrowing occurred shortly
after making contact with the sediment and that the insects did not inspect
sediments prior to burrowing. However, comparison of the behavior of animals
dwelling in sediments from site 4 with other sites (during 1977 tests) show
a greater tendency for Hexagenia to return to the surface after burrowing in
site 4 sediments. Within the 8 chambers, a total of 5 insects returned to the
surface from site 4 sediments whereas, not more than 1 insect returned to the
surface of the sediments from the other sites during the observation period.
Daphnia Bioassays--
The survival of Daphnia suspended over sediments varied from 50-100% with
the lowest survival occurring during the first bioassay. Analysis of variance
suggested that survival of Daphnia was not influenced by oxygen concentration
during the 1977 tests. However, survival was significantly lower during the
first test which was conducted during June 6-10, 1977 (Table 31).
For this first test, survival was low for both the control (Pokegama Slough)
and harbor test site 2 sediment. This low survival in the control suggested
that some condition related to technique rather than sediment quality influenced
the results. Consequently these results were omitted from further analysis.
Comparison of means for the 1977 bioassays by Duncan's multiple range
test (Steele and Torrie, 1960) suggested survival of Daphnia was lower in tests
using sediments from sites 1, 4, 6 and 6R than in water over Pokegama Slough
sediments (Table 31). However, in no individual test were the means from the
-------
-95-
TABLE 31
SURVIVAL OF DAPHNIA MAGNA OVER-HARBOR SEDIMENTS DURING 1977
SURVIVAL %
Number
of
Reps.
Animals
per
Rep.
Low
°2
High
°2
Date
Site
Control
Test
Control
Test
June 6 - June 10
2
8
5
62.5
52.5
55.0
50.0
June 13 - June 17
1
8
5
67.5
85.0
85.0
62.5*
June 27 - July 1
6
8
5
100.0
92.5
85.0
67.5*
July 11 - July 15
3
8
5
90.0
90.0
72.0
65.0
July 18 - July 22
4
8
5
98.0
65.0*
90.0
82.0
July 25 - July 29
5a
8
5
80.0
82.5
95.0
77.1
Aug. 8 - Aug. 12
LS
8
5
82.5
85.0
92.5
95.0
Aug. 15 - Aug. 19
6R
8
5
87.5
62.5*
92.5
72.5
*Means identified as significant (P <0.05) by Duncan's New Multiple Range Test.
-------
test site for high or reduced oxygen conditions both significantly different
from the control site. For site 1, survival was lowest over the test site
sediment at high dissolved oxygen (62.5%), low for Daphnia over Pokegama Slough
sediment at low dissolved oxygen (67.5%) ana highest (85.0%) over site 1
sediment and the control at low dissolved oxygen (Table 31).
To increase precisions observations at different oxygen concentrations
were combined (Table 32). Comparison of means for the 1977-1978 bioassays
(Table 32) by analysis of variance showed time of the test (Blocks) did not
influence survival (P>0.1) but sediment source had a significant effect
(P<0.05). Data from the August 8-12 bioassay was omitted from the analysis
because it compared survival for the two control site sediments (Pokegama
Slough and Lake Superior).
Comparison of results for the individual bioassays established that sur-
vival was significantly lower over harbor sediments from sites 4, 6R (1977
tests) and 3 (1978 test). Tests with sediments from each of these three site
areas were repeated to determine the reliability of the testing procedure.
Although mean survival was substantially different for the tested control
sites for all repeated tests, differences were not identified as statistically
significant for any site twice. Survival results for the 1977 bioassays com-
paring site 3 and 6 sediments with controls showed no statistically significant
differences. However, survival differences were significant for the second
tests using sediment from these sites. In the 1977 test, Daphnia survival
over site 4 sediment was declared significantly lower than survival over Pokegama
Slough sediments. However, mean survival of Daphnia was not significantly lower
over site 4 sediment than over Lake Superior sediment in the 1978 test.
Variation in the results from the repeated test could be related to
variation in the sediments within the site sampled during different time
periods or to the precision of the bioassay procedure. To evaluate pre-
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-97-
TABLE 32
SURVIVAL OF DAPHNIA MAGNA OVER SEDIMENTS 1977-1978
Survival %
Date Site Reps/Test
Value of
Control* Test t
1977
June 13-June 17 1 16 76.3 73.8 0.3
June 27-July 1 6 16 92.5 80.0 1.6
July 11-July 15 3 16 81,3 77.5 0.4
July 18-July 22 4 16 92.5 73.8 2.2
July 25-July 29 5a 16 87.5 81.3 0.7
Aug 8-Aug 12 LS 16 87.5 90.0 0.4
Aug 15-Aug 19 6R 16 90.0 67.5 2.6**
1978
June 12-June 16 4 16 96.2 91.3 1.8
June 19-June 23 3 16 97.5 81.3 4.1 +
* Control represents Pokegama Bay sediments during 1977 and Lake Superior
sediments during 1978.
**Values declared significant with P< 0.05.
+' Values declared significant with P <• 0.01.
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-98-
cision of the procedure, comparisons were made between survival of Daphnia
magna in the two test beakers suspended over each sediment in the test.
Because Daphnia in the two beakers were subject to identical test conditions,
it should follow that survival of animals in one beaker should be positively
correlated with survival in the other. To test this assumption a correlation
analysis was performed employing data from the 1977 bioassays. The calcualted
coefficient based on the entire data set suggested some positive relationship
exists. However the coefficient was not strong or significant (r = 0.16;
P>0.1). Because it was noted that survival was generally high (100-80%),
a second correlation was conducted employing only observations where survival
in one beaker of the pair was 60% or lower. This test was conducted on the
assumption that a strong relationship would indicate the ability of the bio-
assays to identify the presence of toxic effects of sediments where they
occurred. The coefficient suggested a weak negative relationship (r = -0.29
P>0.1). The poor correlation between replicates subject to identical test
conditions, suggests high variability and low precision of the bioassays in
identifying differences in sediment quality.
Chemical Characteristics of Post-Bioassay Water--
The chemical analysis of the water overlying the sediment at the conclusion
of the 96 hr oversediment is presented in Appendix C. The general chemical
characteristics of the overlying water samples do not vary widely for the
various sediments employed in the tests. This result is consistent with the
observation that the animal survival values did not vary widely between sites
and no one sediment site sample caused significant decreases in survival com-
pared to the control for both Daphnia and Hexagenia tests.
Pontoporeia Bioassays —
Average survival of Pontoporeia affinis varied between 27.8 in 100%
elutriate water from test site 4a sediments, to 95.8 in 100% elutriate water
from Lake Superior sediments (Table 33). Analysis of variance was performed
-------
-99-
TABLE 33
SURVIVAL OF PONTOPOREIA AFFINIS IN ELUTRIATE WATERS - 1978
SURVIVAL
No Animals Lake Superior Elutriate Harbor Elutriate
Harbor of per
Site Reps Rep LS 5% 50% 100% 5% 50% 100%
1 4 6 75* 83.3 95.8 83.3 91.7 79.2 87.5
2 3 6 77.8 94.4 50 88.9 66.7 94.4 72.2
3 4 6 87.5 91.7 95.8 95.8 95.8 91.7 95.8
4 4 6 87.5 79.2 91.7 95.8 83.3 91.7 87.5
4a 3 6 77.8 94.4 50 88.9 9.4.4 50 27.8*
5R 3 8 79.2 66.7 95.8 87.5 58.3 91.7 83.3
6 3 8 79.2 66.7 95.8 87.5 66.7 95.8 62.5
*Mean is significantly different (P < 0.05) by Duncan's New Multiple Range Test.
-------
-100-
to compare means from all test sites and the results indicated survival was
not influenced significantly by the source of the sediments from which water
was extracted or concentration of liquid phase elutriate water (P>0.1). In
the analysis, average survival in 5, 50 and 100% elutriate water from Lake
Superior sediments for all bioassays (which was 82.3, 82.1 and 89.7% re-
spectively) was compared with survival in 5, 50 and 100% elutriate water for
each of the harbor sediments tested. The results suggest the test was not
effective in measuring any general differences between toxicity of elutriate
from harbor or Lake Superior sediments. Duncan's new multiple range test was
applied to compare survival of Pontoporeia in Lake Superior water; 5, 50 and
100% elutriate water from lake sediments and; 5, 50 and 100% elutriate water
from 7 harbor sites, for individual bioassays. Although averages varied con-
siderably the test showed survival was influenced significantly with 100%
elutriate water from site 4a and in Lake Superior water during the first bio-
assay (Table 33). Failure of the test to identify more significant differences
may be due to chemical similarity of water from sediments or low precision of
tests in identifying differences. Variation in survival between replicates
was high suggesting precision of the test procedure was low.
ELUTRIATE, INTERSTITIAL AND PORE WATER BIOASSAYS
Methods
Daphnia Bioassay—
Daphnia magna bioassays were conducted with interstitial, elutriate and
generated pore water in 250 ml beakers containing 200 ml of test solution.
Six 96 hr bioassays were conducted during June and July 1977. The 1977 tests
compared survival of Daphnia magna in 5 and 50% interstitial water from the
test sites, survival in 5 and 50% interstitial water from the Pokegama Slough
control site and in Lake Superior water. These tests included 5 replications
arranged in a latin-square design (Steel and Torrie, 1960). During the August
8-12 test, 5 and 50% interstitial water and 50% elutriate water from Lake
-------
-101-
Superior sediments was substituted for interstitial water from a harbor test
site. A second bioassay (August 15-19) also included 50% elutriate water from
site 6 (6R) as a test medium in addition to 5 and 50% interstitial water from
site 6R, the Pokegama Slough control site and Lake Superior water. The August
bioassays included 6 replications arranged in a latin-square design. During
October, bioassays were repeated for sites 3 and 4 (designated as 3R and 4R).
In the October bioassays test media consisted of 5 and 50% interstitial water
and 5 and 501 generated pore water from harbor sediments, 5 and 50% interstitial
water from Lake Superior sediments and Lake Superior water. These tests in-
cluded 5 replications in a completely randomized design.
Tests were repeated for sites 3 and 4 during 1977 because Daphnia magna
survival was reduced by the use of oxygen supersaturated water during the
first test employing sediment for these sites and site 1. During the earlier
test Daphnia came to the surface and died in all the chambers, a condition
apparently resulting from supersaturation of the test water medium. A second
test with sediment from site 6 (6R) was performed to determine the reliability
of the procedure in identifying toxicity of sediments.
Daphnia bioassays were conducted with sediment elutriate water and par-
ticulate phase water in mixtures with Lake Superior water during 1978 to
determine the reliability of the 1977 test results and to identify possible
differences between effects of elutriate, interstitial, generated pore water
and particulate phase water on Daphnia survival. Nine bioassays were con-
ducted with elutriate and particulate phase water from six site areas. The
tests were conducted with Lake Superior water; 5, 50 and 100% elutriate water
from Lake Superior sediment; 5, 50 and 100% elutriate water from harbor site
sediments; and 10% particulate phase water from harbor and Lake Superior sedi-
ments. Each test condition was replicated 5 times.
Fish Bioassays--
Fish breathing response bioassays (Drummond and Carlson, 1977) were con-
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-102-
ducted with sediment interstitial water from Pokegama Slough, Lake Superior
and harbor test sites 1, 3, 4, 5a and 6R. Sixteen fish of comparable size
were used in measuring cough frequency and breathing frequency responses for
each bioassay.
The tests were conducted in a system comprised of three major components
(Figure 5). Four electrode chambers measuring 11 x 14 x 10 cm were constructed
to monitor response from individual fish. Each chamber included four separate
2.5 x 9-12 cm compartments. The linear dimensions of the compartments could
be adjusted by a moveable loose fitting partition, from which 4 stainless
steel wire electrodes were suspended. Electrodes on the moveable partition
and 4 identical ones suspended from the outflow end of the chamber were used
to detect action potential (Heath, 1972) resulting from muscular activity as-
sociated with breathing. During each test, four bluegill sunfish were placed
in each electrode chamber which was suspended in a 3.5 liter (23 x 19 x 14 cm)
glass chamber used to hold the test solution. During acclimation and the
bioassays, dechlorinated city water, interstitial water or elutriate water was
placed in the 3.5 liter chambers and circulated through the grid chambers by
an air lift system constructed of glass tubing. Water lifted from the 3.5
liter chamber into the electrode chamber passed by the moveable partition and
out through a notch in the top of the opposite end of the chambers. Action po-
tentials picked up by electrodes were amplified and recorded by a Gil son IMP-5H
physiograph. The 4 channel physiograph was connected to the electrode chamber
electrodes through shielded cable and a rotating switch. The switch rotated at
a rate of 1 revolution per hour and switched the physiograph amplifiers to a
different electrode chamber every 15 minutes. Switching the changes in electric
potential resulting from fish coughs or changes in opercular activity rates were
recorded for each fish by the physiograph's 4 channel chart recorder.
During each bioassay, bluegill cough frequency and the percentage of time
opercular activity took pi ace,were recorded for fish in dechlorinated city water,
-------
FIGURE: -5
FISH BREATHING RESPONSE BIOASSAY SYSTEM
4-Channel Physiograph
Switching Device
Wire leads
(shielded)
•Wire electrodes
Positive
Airlift Chamber
Incoming
Acclimation >
Water
Teflon
Stopcock
Drain
Hole
Electrode/Baffle
/
H
I
%i1l.i°* rf_ —* * * * *
Air line
Airlift Chamber System
-------
FIGURE: 2
SAMPLING SITES
X
DULUTH
/M
LS
lake superior
SUPERIOR
-------
-104-
10 and 25% interstitial water from harbor test site sediment and 101 interstitial
water from Pokegama Slough sediments. Measurements using dechlorinated water
were made after 2% to 3 days of flow-through acclimation to the water and bio-
assay system. The measurements for dechlorinated city water established base-
line rates under conditions where the air lift system circulated approximately
3 liters of dechlorinated water in the 3.5 liter tank through the electrode
chambers. Baseline measurements were recorded over 2 to 3 hr periods 3 times
during the day following acclimation. After baseline records were made, portions
of the water in the 3.5 liter chambers were removed and replaced with interstitial
water to make up 10 and 25% interstitial water mixtures. Tests were conducted
with one electrode chamber system (4 fish) in 10% interstitial water from
Pokegama Slough, 2 chambers (8 fish) in 10% interstitial water mixture from a
harbor test site sediment and 1 chamber (4 fish) in 25% interstitial water from
the harbor site. Responses were monitored during several (3 or 4) 2-3 hr periods
daily, for three days.
After three days of monitoring response to interstitial water mixtures,
the solutions were siphoned from the 3.5 liter chamber and replaced with a
steady flow of 18°C dechlorinated Lake Superior water. After 2 hrs the flow
of freshwater terminated and breathing response was measured for an additional
2-3 hrs.
Oxygen and temperature of the test solution were measured periodically
during periods when response data were not being recorded. At the end of the
test fish were weighed and measured (Table 34).
Fish coughs consisted of one or more pronounced opercular movements usually
followed by a smaller than normal strength movement and could be identified
as a distinct pattern on the physiograph record. Coughs were counted for 10
minutes of each 15 minute recording period for each fish. Experience with blue-
gills demonstrated opercular activity may be continuous or interrupted by pauses.
The percentage of time (during the 10 minute period analyzed) that opercular
-------
TABLE 34
FISH BREATHING RESPONSE BIOASSAY CONDITIONS DATA SUMMARY
Periods Recorded (15 min/Period)
Date
Site
Fish Length*
cm
Fish Weight*
g
Oxygen
ppm
Temp
Of
Background
Inter-
stitial
Water
Elutriate
W^ter
June 10-14
1
8.6-10.4
9.4-17.2
6.6-7.3
17.9-18.7
6
20
—
July 8-15
3
8.5-10.6
7.7-17.4
—
—
12
20
—
July 18-22
4
8.1-10.2
6.9-14.1
6.4-7.1
18.0-18.5
9
20
—
July 25-29
5a
8.6-10.7
7.1-17.6
9
20
August 8-12
LS
8.3-10.5
8.0-16.7
f* f *"j t*
6.6-/ .6
¦18.1-18.2
10
16
—-
August 15-21
6R
7.4-9.7
4.2-11.5
—
11
21
5
* Measurements made at the conclusion of the tests,
*
0
cn
1
-------
-106-
activity took place was determined by counting the number of mm of recorder
paper on which opercular movement was indicated and comparing to the total
length of the record for the time period.
Results
Daphnia Bioassays--
A latin square analysis was performed on appropriate 1977 data and sug-
gested survival was reduced in interstitial water from sites 5a and 6 (Table
35). Analysis by Duncan's new multiple range test showed survival was lower
in 5 and 50% interstitial water extracted from site 6 harbor sediments than
in Lake Superior water or interstitial water from the Pokegama Slough control
site (Table 35). In bioassays with interstitial water from site 5a, mean
survival in 5% interstitial water differed from survival in Lake Superior water
and interstitial water from the Pokegama Bay site. During the October bioassays,
survival was significantly lower in 5% generated pore water than in 50% gen-
erated pore water, Lake Superior water and interstitial water from Lake Superior
sediments. Survival in 50% interstitial water extracted from site 3R was also
significantly different from survival in Lake Superior water, interstitial water
from Lake Superior sediments or survival in 5% interstitial and 50% generated
pore water.
Analysis of data from the 1978 bioassays by Duncan's new multiple range
test showed mean survival was significantly lower in at least one concentration
of elutriate water from harbor sediment sites 1, 3, 4, 5 and 5R than in Lake
Superior water or Lake Superior elutriate water (Table 36). Survival was also
reduced significantly in particulate phase water from sites 4a and 6. Comparison
with the 1977 results suggests that sites 3, 4, 5 and 6 were identified as having
a significant negative effect on survival during both years. Survival in tests
for site 1 appeared to be lower during 1977 and was significantly lower during
1978 (Tables 35 and 36).
The bioassays failed to show that concentrations of interstitial, elutriate
-------
TABLE 35
SURVIVAL OF
DAPHNIA MAGNA IN
SEDIMENT
INTERSTITIAL
WATER
- 1977
SURV
I V A
L %
Test Elutriate (e)
or Generated
Pore Water (p)
Number
of
Reps,
Animals
per
Rep
Lake
Superior
Control
Interstitial
T - - -
Test
interstitia1
Value
of F
Level
Sign,
Date
C4 4*q
J 1 IC
¦ 5%
50%
5%
Cfl o/
%J UIQ
5% 50%
June 7 - June 11**
2
5
5
16
16
op
0
32
—
—
—
June 14 - June 18
1
3
3
93
93
93
OA
ou
80
—
0.90
NS
June 28 - July 2
6
5
5
68
68
92
16*
28*
--
1A QK
i h . yo
0.01
July 12 - July 16**
3
5
5
12
24
52
4
16
—
--
—
July 19 - July 23**
4
5
5
28
-j 2
44
0
15
--
July 26 - July 30
5a
5
5
93
100
97
52*
84
--
7.97
0.01
Aug. 8 - Aug. 12**
LS
6
5
43
67
60
50
67
77e
--
—
Aug. 15 - Aug. 19
6R
6
5
67
83
80
70
73
57e
—
--
Oct. 17 - Oct. 21
4R
5
£"
0
96
100
96
92
72*
80p* lOOp
—
Oct, 24 - Oct. 28
3R
5
5
100
92
100
100
nn
oU
56p* TOOp
—
--
*Means identified as significant (P < 0.05) by Duncan's New Multiple-Range Test.
**Bioassay results influenced by supersaturation of Lake Superior water with oxygen.
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-108-
TABLE 36
SURVIVAL OF DAPHNIA MAGNA IN ELUTRIATE WATERS - 1978
SURVIVAL %
Lake Superior Harbor Elutriate Particulates.
Elutriate
LS Harbor
5% 50% 100% 5% 50% 100% 10% 10%
1
5
5
96
100
96
96
100
84*
• 92
96
92
2
5
5
96
100
100
100
88
100
100
96
96
2R
5
5
96
100
100
96
100
100
100
96
92
3
5
5
100
100
96
92
88
68*
64*
92
96
4
4
5
96
96
100
96
92
60*
92
96
95
4a
5
5
96
100
100
96
100
88
96
96
80*
5
5
5
96
100
100
100
96
92
66*
96
84
5R
5
5
96
96
100
100
72*
100
88
92
92
6
5
5
96
96
100
100
96
96
100
92
84*
No. Animals
Harbor of per
Site Reps Rep LS
*Mean is significantly different (P < 0.05) by Duncan's New Multiple Range Test.
-------
-109-
or pore water from sediments had a significant influence on survival. Means
based on survival in 5% interstitial water and 5% pore water appeared to have
the same probability of being declared significant as means based on data
for 50% concentrations.
Fish Bioassays--
Cough frequency of bluegills in dechlorinated city water, and 10 and
25% interstitial water mixtures from Pokegama Slough, Lake Superior and five
harbor test sites (1, 3, 4, 5a and 6R) sediments, were generally similar (Table
37). Higher frequencies which occurred with all mixtures during the first
test and with Pokegama Slough 10%*interstitial water during the third test,
apparently resulted from variation between groups and individual fish used in
the test. In addition to variations between fish, cough frequencies varied
considerably in time, with the highest frequencies generally occurring shortly
after addition of the test mixtures to the system. To control variation be-
tween test fish, average cough frequency for each fish in harbor site inter-
stitial water mixtures (12/test) was determined and compared with the back-
ground frequency for the specific fish. Analysis of variance showed that varia-
tion between fish was significant (P<0.01) for several tests (Pokegama Slough,
Lake Superior, site 3). The test demonstrated that cough rates were signifi-
cantly higher than background with interstitial water mixtures from Pokegama
Bay, site 1 and site 4 (P<0.05). Cough frequencies of fish in interstitial
water mixtures from site 5a sediments were significantly lower than background
frequencies (F<0.01). These differences can be attributed to chemical dif-
ferences between test mixtures or observation time.
To control time related variation, average cough frequencies were deter-
mined for peak activity periods for fish in 10%.Pokegama Bay and 10 and 25%
harbor or Lake Superior sediment interstitial water. Generally, peak activity
occurred during the first 24 hours after introducing the test mixtures. The
fish apparently adjusted to the mixtures after 24 hrs resulting in reduced
-------
110-
cough frequency thereafter. Peak frequencies (Table 38) were generally higher
than averages for the 72 hr tests (Table 37). Comparison of peak cough fre-
quency of bluegills in 10% interstitial water from Pokegama Slough with those
for fish in 10 and 25% interstitial water from harbor and Lake Superior sedi-
ments showed cough frequency was higher for site 3, 5a and 6R (Table 38),
Bluegi11 opercular activity occurred almost continuously (91-100%) in
dechlorinated water and 10% interstitial water from Pokegama Slough during most
bioassays. However, bluegills in 10 and 25% solutions of interstitial water
from harbor sites 1, 3, 4, 5a and Lake Superior sediments showed a broken
pattern of opercular movement which included periods of inactivity. Comparisons
between activity of fish in city water with activity in interstitial water
mixtures by analysis of variance showed that the percentage of the time opercular
activity took place were significantly lower for all interstitial water mix-
tures except those from site 6R sediments (P<0.05).
COMPARISON OF RESULTS TO OTHER SEDIMENT BI0ASSAY STUDIES
Prater and Anderson (1976) conducted sediment bioassays using eight sedi-
ments from the Duluth and Superior harbors employing Hexagenia limbata, Arsellus
commam's, Daphnia magna and Pimephales promelus as biological test organisms.
The investigators used an aquarium containing sediment, overlying water and
test animals in conjunction with a continuous water recirculation system.
The results of 96 hr toxicity tests showed that Daphnia magna generally exhibited
the greatest mortality among test organisms in cases where toxic effects were
observed. This observation is consistent with the results presented in our study.
Shuba, Tatem and Carroll (1978) conducted bioassay experiments exposing
aquatic invertebrates to sediments, standard elutriate water and sediment
particulate phases. In these studies, both marine and freshwater organisms
were employed as biological probes. The investigators found few cases of
statisically significant sediment toxic effects compared to controls even though
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-111-
TABLE 37
INTERSTITIAL WATER FISH BREATHING RESPONSE
Average cough rates and percentage of the time fish underwent opercular movement
is identified for dechlorinated water(Background) and interstitial water from
Pokegama Slough (10%)Lake Superior (10+25%) and five harbor sediments (10+25%).
Average Cough Rate
(No/Min)
Opercular Activity Time (%)
Site
Background
Pokegama
Test
Site
Background
Pokegama Test Site
Early
Late
10%
10%
25%
Early
10%
10%
25%
1
2.3
—
2.9
2.7
2.6
99
98
89
80
3
0.8
0.6
0.7
0.7
1.0
91
99
87
53
4
1.1
2.1
3.0
1.0
1.1
100
98
83
68
5a
2.1
0.3
0.8
1.1
1.0
100
69
90
63
LS
0.4
0.8
0.4
0.3
0.8
99
58
56
91
6R
0.2
0.1
0.2
0.2
0.2
97
94
93
92
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-112-
TABLE 38
PEAK COUGH FREQUENCY OF BLUEGILL SUNFISH IN INTERSTITIAL WATER
Peak cough frequency is estimated as the average frequency during the first 22-26
hrs fish were subjected to interstitial water mixtures.
Pokegama
10%
Peak Cough frequencies (no/min)
Test Site
10%
25%
1
2.54
2.62
2.96
3
0.78
1.01*
1.73**
4
1.44
1.09
1.11
5a
1.40
2.10*
2.00
LS
0.28
0.20
0.48
6R
0.13
0.23**
0.15
* Identified as significantly different (P< 0.05).
**Identified as significantly different (P< 0.0T).
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-113-
many of the test sediments contained high concentrations of contaminants such
as metals, PCBs and petroleum hydrocarbons. Their observed mortality results
depended upon sediment quality, test organisms used and the length of time
the sediment was held in the laboratory. In several cases, repeat studies
using the same sediments showing widely different results. This variation
in results was also observed in our work as we did not observe significant
animal toxicity in repeat determinations using sediments sampled from the
same site.
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-114-
SECTION 9
BIOACCUMULATION POTENTIAL OF SEDIMENT ASSOCIATED CHEMICALS
CHEMICAL ANALYSIS OF HEXAGENIA AND CHIRONOMIDS
The potential for benthic organisms to accumulate organic chemicals and
metals in the sediment was investigated by chemical analysis of both bioassay
and naturally exposed animals. Hexaqenia limbata, collected after their use
in the 1978 96-hr bioassay tests (Section 8), and chironomids, obtained in 1977
and 1978 from harbor sampling sites, were utilized in the chemical analysis.
The animals were analyzed for PCBs, certain pesticides, selected PAH compounds
and ten metals.
General Methods
Chironomids collected from the harbor sites (procedure described in Sec-
tion 6) were dried with tissue paper, weighed, wrapped in aluminum foil and
frozen until analysis. Post-bioassay Hexaqenia limbata were washed in distilled
water, dried with tissue paper, weighed, wrapped in aluminum and frozen.
Prior to chemical analysis, Hexaqenia samples were thawed, weighed and dried
overnight at 60°C for subsequent dry weight determinations and metal tests.
For organic analysis, one to two gram samples (wet weight) of chironomids
and two to three gram samples of Hexaqenia were ground with 0.5 ml of concentrated
HC1 using a ceramic mortar and pestle. Anhydrous Na^SO^ was added to yield a
freely flowing mixture which was transferred to a hexane rinsed cellulose
extraction thimble. Each mixture was extracted for 8 hr with 125 ml of a 9:1
mixture (v/v) of hexane-acetone in a Soxhlet apparatus. The extracts were
concentrated to 5 ml in a Kuderna-Danish system employing a three ball Snyder
reflux column. High molecular weight components were removed by gel permeation
chromatography (see Section 7) using methylene chloride as the eluent. For
each sample, the lower molecular weight fraction (<600) was concentrated to
5 ml (Kuderna-Danish apparatus) and further concentrated to':2 ml using a gentle
-------
-115-
stream of ^ blown over the liquid surface. Aliquots (1.0 ml) of the concentrated
extracts were subjected to additional silica gel clean-up and fractionation (see
Section 7) to separate PCBs from pesticides and to isolate the PAH compounds
in the benzene fraction. The analysis of extracts by gas chromatography and
GC-MS (PAH compounds) was carried out by procedures identical to those described
for sediment extracts (Section 7).
Chironomids were prepared for metal analysis by placing about 0.05 g sub-
samples (wet weight) into Parr bombs and adding 2.5 ml of ultrapure HNO^ to
each. The mixtures were digested for one hour at 125°C. After evaporating the
mixtures to near dryness, each was diluted to 25 ml with deionizea water. The
solutions were analyzed by atomic absorption spectrophotometry (Section 7).
Resul ts
The Hexaqenia and chironomid samples contained some inert material in their
digestive tracts since the animals were not placed in clean water for several
days after their removal from the sediments (EPA/Corps of Engineers, 1977).
This inert material would not be incorporated in tissue and could render bio-
accumulation values for tissue either high or low depending upon the concentra-
tions of each chemical constituent associated with the inert material. However
the procedure of placing the animals in clean water for several days presents
some uncertainty in bioaccumulation values due to possible elimination of certain
chemical constituents during this time period.
In order to estimate the amounts of chemicals associated with inert material,
residues remaining after HNO^ digestion of the dried animal subsamples were
weighed. These residues represent inert material which varied from 4 to 13%
of the total dry weight of animal tissue samples for Hexaqenia and 4 to 18%
for chironomids. The actual weights of sediment in the animal samples prior
to digestion in HNO^ were estimated by adding the weight of that portion of the
sediment which dissolved in the digestion process to the residual weight. The
determination of the weights of sediment dissolved in the digestion process was
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-116-
based on values determined in the selective extraction procedure for metal
analysis (Section 7). It had been determined that the residual fraction of
the sediment comprised 80 to 90% of the total dry sediment weight. The measured
concentrations of chemical constituents in the animal samples were corrected
by subtraction of the amounts associated with the sediments (reported in Section
7) and dividing by the fraction of the total sample weight which represented
animal tissue.
Bioaccumulation of PCBs--
Table 39 shows that total PCB concentrations in chironomids from the harbor
sites ranged from 6 to 17 yg/g of dry tissue. The dry tissue weight was es-
timated to be 10% of the wet weight for all samples based on limited measure-
ments. Compared to PCB levels in the dried sediment samples, bioaccumulation
factors of 11 to 18 times were found in the animal tissues. PCB values in
Hexagenia limbata exposed to harbor sediments for 96 hours.were elevated over
the value found for the animals prior to their use in the sediment bioassays
(Ox Creek value). Bioaccumulation of PCBs was also found for Hexagenia in
Lake Superior sediments. The bioaccumulation factors were 4.8 for Hexagenia
in the two harbor sediments and 8.2 in the Lake Superior sediment (dry animal
tissue/dry sediment).
The amounts of PCBs associated with ingested sediment in the animals were
small compared to the total values measured. The corrections for PCBs associated
with ingested sediment were 5% or less for Hexagenia and even smaller for
chironomids. The effect of correcting for the weights of sediments in the animals
was to increase the concentrations of PCBs associated with the animal tissue
by 10 to 20%. Consequently, the PCB values measured for the animals containing
some ingested sediment were low by 10 to 20% compared to the corrected values
given in Table 39.
Bioaccumulation of Pesticides-
Chlorinated pesticides in the sediments were either absent or at very low
-------
•117=
TABLE 39. TOTAL PCBs IN SEDIMENT AND BIOLOGICAL SAMPLES 0978}*
Sediment
Chironomids
Hexaqenia limbata
Site
1
2
3
4
5
6
Ox
Creek
LS
LSH
t
Wet
0.16
0.32
0.64
0.39
0.52
0.39
0.15
Dry
0.31
0.61
1.6
1.0
0.85
0.77
0.33
Dry Magnification Dry Magnification
Tissue Factor Tissue Factor
6
10
17
17
13
9
18
16
11
17
15
12
7.7
4.8
1.7
2.7
4.0
4.8
4.8
8.2
* Concentrations in yg/g.
+ Site 4 Hexaqenia toxicity bioassay control sediment obtained June 14, 1978.
^ Site 3 Hexaqenia toxicity bioassay control sediment obtained June 22, 1978.
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levels, (Section 7) as was the case for the Hexagenia and chironomid samples.
The concentrations of DDT, dieldrin, endrin and heptachlor epoxide were below
detection limits (<10 ng/g of dry tissue).
Measurable concentrations of only p,p'-DDE were found in the chironomid
samples from the harbor sites. These values (in ng/g of dry tissue) were 200
(site 1), 60 (site 2), 140 (site 3), 120 (site 4), 250 (site 4a), and 230
(site 6). Table 11 shows that the p9p1-DDE concentrations in two harbor sedi-
ment samples (dry weight basis) were 9.0 and 3.1 ng/g. Comparing these sedi-
ment concentrations to the values in the animal tissues shows bioaccumulation
of p,p1-DDE in the tissues by factors of about 10 to 40.
Measurable concentrations of p,p'-DDE were found in Hexagenia exposed to
harbor and Lake Superior sediments (1977). These values (in ng/g of dry tissue)
were 470 (site 1), 230 (site 4), 170 (site 5a), 90 (site 6) and 170 (site LS).
Compared to dry weight sediment concentrations (ng/g) of 9 (site 4) and 3.1
(site 6), bioaccumulation factors of 25 and 29 are indicated. For the 1977
Hexagenia bioassay studies, the animals were obtained from Rainy Lake in Inter-
national Falls, Minnesota. The concentration of p,p!-DDE in a sample of these
Hexagenia was about 50 ng/g of dry tissue. The results indicate some bioac-
cumul ation occurred during exposure to harbor and Lake Superior sediments.
Bioaccumul ation of PAH Compounds —
Chemical analysis identified PAH compounds in sediments from sites 4 and
6 (Tables 12 and 13). The presence of PAH compounds in chironomids and Hex-
agenia exposed to sediments from these sites was compared to animals exposed to
Lake Superior sediment.
Extracts from chironomids found in site 6R sediment revealed weak MS ion
intensity signals for three PAH compounds. The ion signals occurred at molecular
weights of 178, 192 and 202. Because of low concentrations, the compounds
could not be accurately quantitated but tentative identifications based on the
computerized library search and concentration upper limits are given in Table
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40. No PAH compounds were found in extracts from chironomids exposed to site
4 sediment.
TABLE 40. PAH COMPOUNDS IN CHIRONOMIDS FOUND IN SITE 6R SEDIMENT.
Retention
Time
(minutes)
MW
Formula
Compound
Concentration
(uq/q)*
11.42
178
C14H10
phenanthrene^
<0.47
13.42
192
C15H12
methylphenanthrene
<0.83
16.08
202
C16H10
fluoranthrene, pyrene
<0.12
* Concentrations expressed on wet weight basis without correction for recoveries
through analytical process.
+ Identified from retention time only.
The organic extracts from Hexaqenia exposed to sediments from sites 4 and
6 showed no presence of PAH compounds. However Hexagenia exposed to Lake Superior
sediment had three PAH compounds identified in their tissue extract. An MS ion
signal for phenanthrene (anthracene) at a molecular weight of 178 and ion signals
for two co-eluting isomers of methylphenanthrene were indicated. The source
of these PAH compounds is uncertain since they were not detected in the Lake
Superior sediment. No PAH compounds were found in an extract of Hexaqenia
from Rainy Lake (International Falls, Minnesota). Animals from this location
were used in the 1977 Hexagenia bioassays. Table 41 summarizes the PAH results
for Hexagenia exposed to Lake Superior sediment.
If these upper limit wet weight concentrations for chironomids are converted
to dry weight concentrations based on the estimate that 90% of the total weight
of the animals is due to water, upper limit dry weight concentrations of 4.7,
8.3 and 1.2 yg/g are obtained for phenanthrene, methylphenanthrene, and fluoranthrene
(or pyrene), respectively. Comparison of these concentrations with the cor-
responding dry weight sediment concentrations of 0.72, 0.27, and 1.26 (0.85)
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(Table 13) yield upper limit accumulation factors of 6.5, 30.7, and 1.0
(1.4) for phenanthrene, methylphenanthrene, and fluoranthrene (pyrene), re-
spectively. In comparison, it has been found that fluoranthene accumulated
in oysters (Crassostrea virqinica) 694 and 10,000 times after 2 ana 8 days
of exposure, respectively (Lee e_t al_., 1978).
TABLE 41. PAH COMPOUNDS IN HEXASENIA EXPOSED TO LAKE SUPERIOR SEDIMENT.
Retention
Time Concentration
(minutes)
MW
Formula
Compound*
(w3/q) +
11.34
178
C14H10
phenanthrene
<1.38
13.34
192
C15H12
methylphenanthrene
<0.27
13.52
192
C15H12
methylphenenthrene
—
* Due to weak ion intensity signals, computerized library search was not used
but tentative identifications were made from molecular weights and retention
time.
Concentrations expressed on wet weight basis without correction for recoveries
through analytical process.
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Bioaccumulation of Metals —
The concentrations of metals in chironomids collected from sediment at the
various harbor sites are shown in Table 42. Based on the residues remaining
undissolved after treatment of the animal samples with concentrated HNO^ in
Parr bombs, it is estimated that large percentages of some of these metals were
associated with inert material such as the residual phase of sediment particles.
More than 25% of the measured concentrations of arsenic, lead, manganese and
nickel was estimated to be contained in inert material for at least three
different animal samples. In particular for lead, most of this metal was associated
with inert material for at lease 5 of the site samples. Consequently, much of
the total amount of these metals, as measured in the chironomids, would not be
available for bioaccumulation in predators.
TABLE 42. METALS IN CHIRONOMIDS FROM DULUTH-SUPERIOR HARBOR SEDIMENTS.*
Site+
As
Cd
Cr
Cu
Fe
Pb
Mn
Ni
Zn
Hg
1
0.6
3.5
4.2
51
1250
11.0
54
25
110
3.8
2
3.2
1.5
11.8
54
11800
3.2
233
40
200
0.9
2R
3.7
4.2
7.2
106
6250
5.6
119
36
58
0.4
3
1.0
1.2
6.5
38
3830
3.2
105
9
500
1.8
4
1.4
0.7
8.9
53
10580
b .3
159
29
152
0.7
5a
1.2
2.2
7.0
35
7120
5.0
146
13
46
4.2
6
2.9
2.0
5.6
35
4840
4.3
547
10
143
9.0
6R
2.8
0.4
10.9
31
11260
18.5
231
14
121
1.6
* Concentrations are yg of metal per gram of wet tissue.
"f"
Chironomids were collected from sites 2, 3, 6 and 6R during the spring of 1977.
The chironomids from sites 2R, 4 and 5a were collected during the fall of 1977.
The chironomids from site 1 were a combined sample from spring and fall collections.
The concentrations of the metals in the chironomids did not show positive cor-
relations with metal concentrations in sediments from the sites. However
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the sediments exhibited considerable heterogeneous composition at each site
and the chironomids were collected from numerous dredge samples at these sites.
This presents some uncertainties as to the sediment metal concentrations which
were representative of material in contact with the animals. In addition, the
levels of metals in the animals may show seasonal variations as indicated by
the values for sites 2 and 2R (a repeat sampling of site 2).
The degree of bioaccumulation of metals by the animals from sediments is
difficult to assess. However, it is likely that the observed concentrations of
mercury, chromium and cadmium were accumulated in the tissues of the chironomids
since the amounts associated with ingested sediment were small.
Related Studies
Our results indicated a bioaccumulation of PCBs and DDE by chironomids and
Hexagenia compared to levels in the sediments. Zitko (1974) found that fish
accumulated Aroclor 1254 in their tissues upon exposure to suspended silica
particles containing the PCB mixture. Courtney and Denton (1976) observed that
the hard-clam (Mercenaria mercenaria) accumulated Aroclor 1254 adsorbed on the
surface of alumina particles. In both of the above studies, it was found that
the lighter chlorinated PCB isomers in the 1254 mixture were preferentially
accumulated in the tissues.
Nathans and Bechtel (1977) found that some of the DDT adsorbed on artifi-
cial sediments was available for uptake by deposit feeding annelids. Peddicord
and McFarland (1978) conducted chemical uptake tests using blue mussels (M.ytilus
edulis), coast mussles (M.ytilus californianus)» soot-tailed sand shrimp (Crangon
nigromaculata) and dingeness crabs (Cancer magister) exposed to contaminated
sediment in marine systems. The investigators found that only Mytilus
californianus took up DDT or its metabolites in the form of DDE. None of
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these species accumulated PCBs. Studies on uptake of metals indicated slight
accumulation of As, Cu, Fe, Pb and Zn from suspensions of contaminated sedi-
ment by Mytil us edulis and As, Cd, Cu, Zn, Mn and Ni by Crangon nigromaculata.
Neff et al_. (1978) reported accumulation of Cd, Cr, Cu, Fe, Mn, Ni, Pb
and Zn by Ranqia cuneata, Palaemonetes pugio, Palaemonetes kadiakensis, Neanthes
arenaceodentata and Tubifex sp. from sediments during exposures up to six weeks.
Some of these species also showed accumulation of Hg and V from sediments. The
investigators found variability in metal uptake according to species, salinity
of the water, and possibly season. No correlations held between observed bioac-
cumulation and bulk metal content of the sediment. The authors have reviewed
other studies of accumulation of heavy metals from sediments.
Namminga and Wilhm (1977) investigated heavy metals in water, sediment
and chironomids from a creek in Oklahoma. They presented evidence that copper
and zinc were accumulated in the chironomids compared to water or sediment.
Concentrations of 1.91 and 57 ug/g of Cu and Zn respectively were found in
chironomids. These values represent bioconcentration factors compared to sedi-
ments of 1.1 for Cu and 3.6 for Zn. In contrast, mean concentrations of chromium
and lead were lower in the chironomids than found in the sediments.
SAMPLE SCREENING FOR ORGANIC CHEMICALS USING HPLC
A wide variety of natural and industrial orgainc chemcials may be associated
with sediments since the partition coefficients pertaining to their distribution
between sediment particles and water are often large. The assessment of the
amounts of industrially derived organic chemicals associated with sediments
is difficult because of their potentially large number and variety. A com-
prehensive survey results in high costs and the necessity of using a number
of analytical techniques. Chemicals of primary concern are those with high
lipid solubility (such as PCBs) since these organic compounds tent to bio-
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accumulate In aquatic animals. The magnitude of the lipid solubility of an
organic chemical is related to its bioconcentration in aquatic organisms
(Carlson et'al_. 1975),
Recent studies have demonstrated that the retention time of an organic
chemical on a reverse-phase column, as measured using high pressure liquid
chromatography (HPLC), correlates with its n-octanol/water partition coefficient
(Veith and Austin, 1976; Veith and Morris, 1978; Veith et al_. 1979). The lo-
garithm of the retention time was found to be linearly related to the logarithm
of the partition coefficient (P) for a large variety of compounds whose lipid
solubility properties varied by six orders of magnitude.
The objective of this phase of our study was to investigate the utiliza-
tion of reverse phase liquid chromatography in screening extracts from sedi-
ments and from aquatic animals exposed to the sediments for organic compounds
with high lipid solubility.
Methods and Samples
The liquid chromatography system at the Environmental Research Laboratory,
Duluth, Minnesota was used. This system has been described elsewhere (Veith
and Morris, 1978; Veith et_ al_., 1979). In summary, it consisted of a Varian
4200 instrument employing two 5000-psi pumps, a high-pressure stopflow injector,
a fixed wavelength UV detector (254 nm) equipped with a 8-ul flow cell of 1 cm
(r)
path length and a Varian Micropakw C-10 analytical reverse phase column (250 mm
x 2 mm i.d.). The detector was interfaced with a Hewlett Packard 3354 mini-
computer for retention time and peak area determinations.
The column was maintained at 50°C during operation. Chemicals under in-
vestigation were contained in a 3:1 mixture (by volume) of acetone and cyclo-
hexane (standards used to calibrate column) or hexane (aquatic animal extracts
-------
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and sediment extracts). Gradient elution was employed by using a solvent
system initially consisting of 30% methanol in water and increasing linearly to
1001 methanol at the rate of 2%-min-1. Flow rate was 2.0 ml•min"1 at 3000 psi.
Chemicals move through the column according to their partitioning character-
istics and elute in the order of least hydrophobic to most hydrophobic. Con-
sequently those chemicals with the greatest bioaccumulation characteristics
(high lipid solubility) were eluted with the longer retention times. Chemicals
with log P values ranging from 0.6 to 6.7 were separated and eluted in about
30 minutes employing this gradient elution procedure.
The elution times of chemicals were correlated to their log P values by
injecting a mixture of benzene, bromobenzene, biphenyl, bibenzyl, p,p'-DDE and
2,4,5,2' »5'-pentachlorobiphenyl. The components of this mixture had log P values
of 2.13, 2.99, 4.09, 4.81, 5.69 and 6.11, respectively. Retention times were
computed relative to the retention time of phenol which was used as an internal
standard.
Portions (20 p£) of the extracts from sediments, Hexaaenia limbata and
Chironomids (Section 7) were passed through the liquid chromatograph. The
general operating conditions of the chromatograph were the same as described
above. The retention times, peak areas and log P values of eluting compounds
were tabulated by the minicomputer. If a retention time matched that of a
compound in the computer memory within ± 5%, the tentative identity of the com-
pound was printed along with its retention time.
Results
It was not possible to quantitate compounds by monitoring the UV absor-
bance of eluting components in the mixtures. The absorptivities of organic
compounds at 254 nm can vary by orders of magnitude. Since the eluting com-
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pounds were not positively identified they could not be quantitated. Also any
compounds with low absorptivities at 254 nm would not be detected. Consequently
the only comparisons made between the samples were the number of strong, medium
and weak, elution peaks at various log P values.
Table 43 summarizes the number of elution bands observed in various log P
ranges for sediment, chironomid and Hexagenia samoles. The elution bands are
qualitatively classified as to being strong (s), medium (m) or weak (w) in terms
of relative absorbance at 254 nm.
Comparison of the elution bands for sediment extracts revealed the largest
number occurring at log P values greater than 4. Site 2 shows the fewest elu-
tion bands with the other sites exhibiting similar numbers. The strongest ab-
sorption band in sediment extracts from four of the sites and the second strong-
est absorption band in extracts from the other two sites occurred at log P = 5.7
which corresponds to the log P value for DDE and also benz(a_)pyrene. However
the identity of the compound(s) giving rise to this band was not determined.
The chromatograms of the animal extracts show larger numbers of low log P
elution bands than sediments corresponding to relatively more polar compounds
in the animals. However fewer elution bands with log P>4 values were observed
than found for sediment extracts. The strongest absorption band for all the
chironomid extracts occurred at log P = 1.5 which may be a naturally occurring
compound in the animals. The second strongest absorption band was found at
log P =4.7 which corresponds to the value for bibenzyl. However many other
compounds could cause this band. For the Hexagenia extracts, site 2 showed
the least number of elution bands similar to the situation for sediments. The
strongest absorption bands occurred at log P = 1.4 (sites 5 and 6), log P = 2
(site 2) and log P = 3 (site LS).
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TABLE 43. SAMPLE ELUTION BANDS OBTAINED BY REVERSE PHASE CHROMATOGRAPHY.
Number of Eluted Bands*
Site Sample Type
0 <
log P
< 2
2 <
log P
< 4
4 <
log P
< 6
log
P
> 6
S
M
U
S
M
W
S
M
W
S
M
W
1 Sediment
0
0
4
0
0
1
1
3
5
0
5
2
Chironomids
1
2
3
1
2
2
2
1
1
0
0
0
2 Sediment
1
0
4
0
1
0
2
2
3
0
0
3
Hexagenia
2
1
4
1
1
1
0
0
0
0
0
2
3 Sediment
0
0.
1
0
0
2
3
6
0
0
3
6
Chironomids
1
2
5
0
1
5
3
1
0
0
0
3
4 Sediment
0
3
4
0
0
4
2
2
7
2
3
3
5A Sediment
0
1
3
0
0
2
2
6
0
1
2
4
Chironomids
2
1
4
1
1
0
5
0
0
0
1
1
Hexagenia
3
1
9
1
3
4
1
4
3
0
0
3
6R Sediment
0
2
3
0
1
2
5
4
2
1
2
3
Chironomids
1
0
3
2
1
2
5
2
1
0
0
0
Hexagenia
2
8
7
1
4
2
1
0
3
0
3
5
LS Hexagenia
1
2
5
2
3
3
2
4
1
0
0
3
* W, M and S refer to weak, medium and strong respectively. The relative in-
tensity ranges used in these classifications are W (0.1 to 1.0), M (1.0 to
3.5) and S (3.5 to 80).
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Discussion
The use of reverse phase liquid chromatography shows potential as a useful
method in screening environmental samples for chemicals with high lipid solu-
bility. However the large diversity of organic chemicals which may be associat-
ed with the sediment particles presents some difficulty in quantitating their
amounts. Eluent fractions from various log P ranges could be collected and
combined with gas chromatographic and/or mass spectroscopic techniques for
identification and quantitation. This procedure would greatly increase the
time and expense required in the screening process. Another possibility is to
use a fluorescence detector in series with the UV detector for additional moni-
toring capabilities such as screening for specific PAH type compounds.
Since many industrial organic compounds contain halogens, a determination
of such compounds (particularly with higher log P values) would be valuable.
This determination might be accomplished by collecting elution fractions in
various log P ranges and subjecting these fractions to halogen specific analysis.
For example, Glaze et_ al_. (1977) used a microcoulometric technique to measure
total halogen content of effluents at nanogram levels. The samples screened
in this manner could consist of sediment extracts or extracts from benthic
organisms after a certain sediment exposure period.
Another possible method which might be employed to arrive at an index of
the organic compounds with high lipid solubility in sediments would involve a com-
bined chromatographic and gravimetric procedure. Extracts from sediments or
animals exposed to sediments would be cleaned-up by gel permeation chromatography
and then chromatographed on a preparative reverse phase column using the method
described in this section but with larger injection amounts. After compounds
with log P values less than three or four were eluted, the solvent would be
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changed to pure methanol to remove the rest of the organic compounds (those
with high log P values) from the column. This methanol eluent would he col-
lected In tared crucibles and evaporated. The remaining residue would be
weighed as a measure of total bioaccuable material. The residue could be
subjected to further analysis aimed at compound identification and quantitation
if desired. Use of a 20 g sample containing 10 yg/g of high log P compounds
would yield 0.20 mg of residue. Consequently, careful microgravimetric methods-
would be required.
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SECTION 10
RELATIONSHIPS BETWEEN CHEMICAL CHARACTERISTICS
OF THE SEDIMENT SYSTEMS AND BIOASSAY TOXICITY
SITE LOCATION AND TOXICITY
The complex and variable nature of the sediments and water systems derived
from them (interstitial, elutriate and generated pore water) reduced the likely-
hood that chemical measurements adequately reflected the complicated chemical
conditions in the bioassay test chambers or that a single chemical constituent
was responsible for the observed toxicity. Furthermore, variation in survival
rates between replicates subject to identically prepared sediment-water systems
indicated precision of the bioassay tests were low. Recognizing these limitations,
emphasis was placed on development of a general index of toxicity and chemical
quality of harbor sediments in trying to identify relationships between sediment
quality and animal survival. Because it is possible that some chemical parameters
could serve as an indicator of toxicity, an effort was also made to identify
correlations between concentrations of specific chemical constituents in the
sediments and survival in the bioassay.
Based on the assumption that industrial development within the harbor re-
presents the primary source of toxic substances in the sediments, location of
the sampling sites was considered to be a potentially strong indicator of overall
sediment toxicity. Application of this simple indicator presented an alternative
method to using sediment chemical characteristics to predict toxic effects since
the chemical tests may not measure certain toxic substances or the variable
toxicity of identified constituents according to their chemical speciation.
A general index of toxic effects was developed for each site from the total
number of significantly lower mean survival measurements. The total was divided
by the number of conducted site bioassay tests to determine the relative per-
centage of low survival for each site. These percentages are presented for each
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site in Table 44, and show survival was generally lower in tests representing
bioassay systems derived from sediments from sites 3, 4, 5, and 6 (including
3R, 4a, 5a, 6R), which are located in the industrialized areas of the harbor.
The percentages show survival was higher in those areas of the harbor which are
undeveloped (Pokegama Slough) those influenced by the lake (site 1 and 2) and in
the lake proper.
The results suggest that the battery of bioassay tests provide a good gen-
eral index of harbor sediment quality. The fact that tests employing Daphnia,
constantly showed low survival for at least 3 of the 4 industrialized sites
indicates Daphnia is the most sensitive among the test organisms employed.
Because bioassays with Pontoporeia were the only tests in which survival in Lake
Superior water was low, this suggests that the Pontoporeia bioassay results are
not reliable.
GENERAL SITE CHEMISTRY AND TOXICITY
To determine whether sites could be generally characterized as to toxic
effects from the chemical analysis, sediments from the test sites were ranked
on the basis of the results of chemical analysis performed on the sediments
or on interstitial water extracted from the sediments. The 7 to 9 sites were
assigned ranks from one (representing the site with the lowest concentration
of a chemical parameter) to the number of sites considered (representing the
site with the highest concentration of the parameter) on the basis of the
following:
1. Sediment concentrations (except residual phase metals) of COD, NH^,
TKN, Total S, Total P, Oil and Grease, Total Hg, Pentachlorophenol,
As, Cd, Co, Cu, Fe, Pb, Mn, Ni, Se and Zn.
2. Interstitial water concentrations of COD, H^S, NH^, TKN, Total Hg,
Total PCBs, As, Cd, Cr, Co, Cu, Fe, Pb, Mn, Ni, Se and Zn (1977 data)
or As, Cd, Cr, Co, Cu, Fe, Pb, Mn, Ni, Se and Zn (1978 data).
-------
TABLE 44. BIOASSAYS FOR A SAMPLING LOCATION WITH THE NUMBER OF LOW SURVIVAL ESTIMATES DECLARED SIGNIFICANT.
Bioassay Test with Number of Significant Results
Oversediment* Interstitial** Elutriate and Pore Water*
Sampling
Site
Number
of Tests
Hexagenia
Daphnia
Daphnia
Lempomi s
Liquid Phase
Daphnia Pontoporeia
Particulate
Daphnia
Total
Percent
1
6
—
0
0
0
1
0
0
1
17
2,2R
6
0
0
0
0
0
0
0
0
3,3R
10
0
1
0
2
3
0
0
6
60
4 5 4a,4R
13
1
1
1
0
2
1
1
7
RA
5,5as5R
9
0
0
1
1
2
0
0
4
44
6,6R
10
0
1
2
1
0
0
1
5
50
LS
40
0
0
0
0
0
1
0
1
3
Pokegama
28
1
0
—
0
—
1
4
* Based on data from Tables 30 and 32.
**Based on data from Tables 35 and 38.
* Based on data from Tables 33 and 36.
i
CO
ro
i
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The general chemical rankings were arrived at by totaling ranks for each
parameter and dividing by the number of parameters considered. Regression analysis
showed that ranks assigned on the basis of sediment chemistry were not highly
reliable in predicting ranks based on intersitital water analysis (Slope = 0.3174;
P>0.1). Variation between ranks assigned on the basis of sediment and interstitial
water chemistry may result from differential solubility of each chemical in sedi-
ments and because concentrations of many parameters were similar for sediments
or interstitial water from the sites, reducing the reliability of the ranking.
"Rankings based on sediment chemical analysis indicated sediments from the
industrialized areas of the harbor (sites 3,4,5,6,6R) were more chemically per-
turbed than Lake Superior sediments or those from harbor sites 1 and 2 which
are located in less developed areas and are strongly influenced by the lake (Table
45). Ranks based on the 1977 sediment chemistry showed the strongest correlation
with relative bioassay low survival percentages (Table 44) for each site (r = 0.80;
P>0.1). In calculating the correlation, ranks for sites 6 and 6R were averaged.
Correlations between relative bioassay low survival percentage and site rankings
based on the 1977 and 1978 interstitial water analysis or the average of ranks
based on sediment chemistry and interstitial water analysis were not as strong
and were not significant (P>0.1).
The analysis suggests that chemical measurements represent a fair predictor
of general toxicity and that chemical analysis of sediment quality may be a
better indicator of toxicity than interstitial water chemistry.
SPECIFIC CHEMICALS AND TOXICITY
Daphnia was found to be the most sensitive among the animals employed in
the bioassay and extensive chemical analysis was performed on Daphnia bioassay
systems. For these reasons analysis of relationships between concentrations
of specific chemicals and toxicity was based upon the Daphnia bioassays. The
bioassay system for which the greatest amounts of related chemical data were
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TABLE 45. SITE RANKING ACCORDING TO SEDIMENT AND INTERSTITIAL WATER CHEMISTRY.
RANKING
1977 1978
Interstitial Interstitial 1977-1978
Site Sediment Water Average Water Average Percent*
1
3.05
3,25
3.20
3.22
3.2
17
2
2.32
4.76
3.54
3.78
3.7
0
3
5.32
5.76
5.54
5.00
5.3
60
4
6.89
4.82
5.86
3.78
4.8
54
4a
___
4.86
4.9
5a
4.21
3.23
3.72
—
3.7
44
6
4.84
5.88
5.36
3.89
4.6
50
LS
4.11
5.44
4.77
3.22
4.0
3
* Percent of low survival estimates declared significant (Table 44).
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obtained was the system employing Daphnia magna suspended above sediments within
bioassay chambers. Consequently, the results from this test system were used to
compute correlation coefficients between survival results and concentrations of
specific chemical parameters.
The survival values for Daphnia magna over harbor sediments (1977 tests)
were given in Table 31. These survival data for Daphnia at high D.O. and low
D.O. conditions in Lake Superior water over harbor sediments were used for test-
ing correlations with sediment and water chemistry.- Only the survival data per-
taining to experiments in which the control Daphnia survival was greater than
80% were used. Student-t values were computed for testing significant differences
between mean Daphnia survival above the sample and above the reference sediment
(8 replications for each sediment). Six t values were obtained under both high
D.O, and low D.O. conditions. The t values, for each set of D.O. conditions,
were correlated to sediment chemistry (Table 2, Tables 4 through 9 and Table 11
(Total PCBs) ), to interstitial water chemistry (Table 16), to elutriate water
from sediments exposed to oxygen (Table 18). Those chemical parameters which
showed significant correlations (r exceeds 0.81) are summarized in Table 46.
The correlations results should be considered in conjunction with the un-
certainties in the chemical parameters and the limited number of sediment sites
tested. Some chemical parameters are near the limits of detection and thus real
variation trends are less certain. Other parameters tend to show relatively
high fractional standard deviations upon replicate analysis. Even with these
considerations, it appears that manganese values may be a good indicator of po-
tential toxicity toward Daphnia in water above the sediments. Positive correlations
were found for Daphnia toxicity and manganese concentrations in a number of the
sample types (interstitial water, oxic elutriate water, sediment organic phase
and total sediment concentration). Larger amounts of manganese in the interstitial
water could also indicate the presence of larger amounts of other metals which
were associated with Mn(0H)3. However positive correlations of other interstitial
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TABLE 46. CHEMICAL PARAMETER-TOXICITY CORRELATIONS FOR DAPHNIA MAGNA IN WATER
OVERLYING SEDIMENTS
Oxygen
Chemical
Conditions
Correlation
Sample
Parameter
in Water*
Coefficient
Sediment
COD
low
0.83
Sediment
NH-
Total Sulfide
high
-0.89
Sediment
low
0.85
Sediment (organic phase)
Mn
low
0.88
Sediment (organic phase)
Mn
high
0.82
Sediment (residual phase)
As
high
-0.90
Sediment (residual phase)
Ni
low
0.82
Sediment (total metals)
Co
low
0.85
Sediment (total metals)
Cu
high
0.85
Sediment (total metals)
. Fe
low
0.81
Sediment (total metals)
Mn
low
0.88
Sediment (total metals)
Zn
low
0.81
Interstitial Water
Cd
low
-0.96
Interstitial Water
Co
high
0.90
Interstitial Water
Mn
high
0.85
Interstitial Water
Ni
low
-0.94
Interstitial Water
Se
high
-0.84
Interstitial Water
CI
high
0.86
Interstitial Water
Elutriate Water (oxic)
Total PCBs
high
-0.88
NH.
low
0.83
Elutriate Water (oxic)
As3
low
0.84
Elutriate Water (oxic)
Cd
low
0.82
Elutriate Water (oxic)
Cr
high
0.86
Elutriate Water (oxic)
Pb
low
0.82
Elutriate Water (oxic)
Mn
low
0.81
Elutriate Water (anoxic)
pH
low
-0.84
Elutriate Water (anoxic)
Total phenols
high
-0.81
Elutriate Water (anoxic)
Arsenic
low
0.87
Elutriate Water (anoxic)
Total PCBs
high
-0.95
*Low and high dissolved oxygen values were 1 to 5 mg/1 and 6 to 9 mg/1 respect-
ively.
4*
Oxic refers to sediment exposed to air prior to elutriate water preparation
while anoxic elutriate was prepared from sediment not exposed to air.
-------
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water metals with Daphnia toxicity were not found except for cobalt. Interestingly,
a number of metals show positive correlations according to their total concentrations
in the sediment.
Relationships were investigated between the concentrations of individual
metals in elutriate waters (1978 tests; Table 21) and the survival of Daphnia
magna in these waters (Table 35). The percent survival of the animals in the three
elutriate water-Lake Superior water mixtures (5%, 50% and 100% elutirate) were
compared to survival in corresponding elutriate water control (prepared using
Lake Superior sediment) by computing student t values. The t values for the 5%
mixtures for all sites (nine t values) were tested for correlations to concentra-
tions of individual metals in both filtered and unfiltered elutriate water.
Similar correlations were tested between metals and Daphnia survival for the
50 and 100% elutriate water mixtures.
Interstitial water from the sediments used in the 1978 tests was also analyzed
for certain metals (Table 17). These metal concentrations pertaining to either
filtered or unfiltered interstitial water were tested for correlations with the
t values in the same manner as described using metal concentrations in elutriate
water.
Significant correlations (r>±0.71) were found between cadmium concentration
in unfiltered elutriate water and t values for Daphnia survival in 5% elutriate
water-Lake Superior water mixtures (r = -0.72) and between copper concentrations
in filtered elutriate water and t values for Daphnia survival in 5% elutirate
water-Lake Superior water mixtures (r= 0.71). However, no significant correla-
tions were identified for tests with 50 and 100% elutriate water mixtures suggest-
ing the instances of correlation may be fortuitous.
More correlations were found between the t values for Daphnia survival and
interstitial water metal concentrations. In the case of iron in interstitial
waters, r values of 0.80, 0.63 and 0.49 (5%, 50%, 100% elutriate water respectively)
-------
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were found for unfiltered interstitial water while r values of 0.62, 0.88 and 0.78
were computed for filtered interstitial water. Since three of these values are
significant and all are positive, the concentration of iron in the interstitial
water may be an indicator of toxicity toward Daphnia in elutriate water-Lake
Superior water mixtures. Other significant correlation coefficients were found
for arsenic (501 mixture, r=0.78), chromium (50% mixture, r=0.74), nickel (50%
and 100% mixtures, r=0.97 and r=0.80 respectively) in unfiltered interstitial
water.
-------
-139-
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APPENDICES
A - Analytical Methods of Analysis ................. 146
B - Analysis of PCB Gas Chromatographic Elution Patterns . 154
Identification of PAHs ..... ......... 159
C - Chemical Characteristics of Duluth-Superior Harbor Water
(Table C-l), Lake Superior and City of Superior Water
(Table C-2), and Hexagenia Bioassay Water at High D.O. and
Low D.O. (Tables C-3, C-4, and C-5). 174
D - Great Lakes Harbor Sediment Evaluation Criteria (Jensen
Criteria). . 182
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APPENDIX A
ANALYTICAL METHODS OF ANALYSIS
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ANALYTICAL METHODS OF ANALYSIS
A. Sediment Analysis
The data for the sediment core analysis is given in Tables 2, 3,
9 and 10. The procedure used for analysis of each parameter is described
below.
1. Eh_. The Eh values for the sediments were measured by inserting a
platinum and a silver-silver chloride electrode (4M KC1) into the
sediment core to a depth of 2 cm. The millivolt display on the
Corning Model 5 pH Meter was read and corrected to a scale corres-
ponding to use of the standard hydrogen electrode as the test
electrode.
2. joH_. The pH of the sediments was obtained by insertion of a glass
and a silver-silver chloride electrode two centimeters into the
sediment and reading the value from a Corning Model 5 pH Meter.
3. Particle Size Analysis. Pretreatment of the sediment was done to
remove soluble salts and organic material. The pi pet-sedimentation
method was used to measure dispersed particle settling rates (Royse,
1970). The sample was washed through a #230 mesh sieve into a
cylinder using an aqueous solution of a dispersing agent (sodium
hexametaphosphate), Aliquots of the suspension were removed from
the cylinder at specified distances below the surface as a function
of time. The weights of particles in the aliquots were determined
by drying them at 105°C and related to the particle size distribu-
tion in the sample by using Wadell 's practical sedimentation formula
tables (Pezzetta, 1972).
4. Chemical Oxygen Demand. A weighed portion of sample was preserved
by adding distilled water and sulfuric acid. The sample was re-
fluxed with potassium diehromate in the presence of mercuric sul-
fate, silver sulfate and sulfuric acid for 2 hours. Unreacted
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dichromate was titrated with standardized ferrous ammonium sulfate.
The volume of titrant used for the sample was corrected for di-
chromate loss utilizing reagent blanks carried through the same
digestion and titration procedure (Fuller, 1969),
5. Ammonia Nitrogen. A preweighed sample, which was preserved with
sulfuric acid, was washed into a Kjeldahl flask and the pH was
adjusted to approximately 6.6, The sample was distilled and the
distillate was collected in 0,02 N sulfuric acid (Fuller, 1969),
The collected distillate was analyzed by adding sodium hydroxide
and determining the amount of ammonia produced with an ammonia
gas sensing electrode (APHA, 1975).
6. Kjeldahl Nitrogen. The sample was digested with sulfuric acid,
sodium sulfate and mercuric sulfate for 30 minutes following the
appearance of sulfur trioxide fumes. The solution was cooled, water
and a 501 sodium hydroxide-sodium thiosulfate solution were added
making the digestate alkaline. The resulting solution was distilled
into a 2% boric acid solution to collect the distillate. The
collected distillate was analyzed using the same procedure employed
for ammonia analysis (Fuller, 1969).
7. Total Solids. The weighed sediment samples were dried in crucibles
overnight at 103 to 105°C and reweighed. The change in weights
were used to calculate total solids.
8. Volatile Solids. Volatile solids were determined by placing the
oven-dried crucibles from the total solids procedure in a muffle
furnace at 600°C for one hour. The change in weight of residue
after ashing was used to calculate the volatile solids.
9. Total Phosphorus. A sediment sample was digested with a mixture of
sulfuric and nitric acids. After digestion the sample was cooled,
water was added and it was made slightly basic with sodium hydroxide.
-------
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The resulting orthophosphate was determined colorimetrically
using the stannous chloride method (APHA, 1975).
10. Oil and Grease. A preweighed sediment sample was acidified with
hydrochloric acid to an approximate pH of 2. The acidified sample
was then dried by grinding it with magnesium sulfate. The dried
sample was placed in an extraction apparatus and extracted for 4
hours with hexane. The hexane was distilled off and the flask was
cooled and dried in a dessicator and weighed. The difference in
the final and initial weights of the extraction flask were used
to calculate the amount of oil and grease in the original sample
(Fuller, 1969).
11 . Total Sulfide. A sediment sample, previously preserved with zinc
acetate, was placed in a distillation apparatus and flushed with
nitrogen. Hydrochloric acid was added to the sample to make it
acidic. The sample was distilled and the distillate was collected
in 0.2 N zinc acetate. Color development was achieved by adding
an amine solution and iron (III) chloride to the distillate. The
sulfide concentration was determined by comparing the color developed
in the sample distillate to that of standards and a blank that were
carried through the same procedure (Fuller, 1969).
12. Total Phenols. Sediment samples to be analyzed for total phenols
were preserved by adding copper sulfate and phosDhoric acid. The
preserved samples were diluted to 500 ml with deionized water and
distilled to remove the phenols from any nonvolatile impurities.
Ammonium chloride was added to the distill able phenols and the pH
was adjusted to a value of 10 with ammonium hydroxide. Aminoantipyrine
and potassium ferricyanide were added to develop a yellow color.
This solution was extracted with chloroform and the extract was
analyzed colorimetrically (APHA, 1975).
-------
-150-
13. Total Mercury. A one to two gram sample was digested with a mixture
of nitric acid, sulfuric acid, a 6% solution of potassium permanganate
and a 5% solution of potassium persulfate. The sample was cooled
and diluted to volume with deionized water. A sodium chloride-
hydroxylamine sulfate solution (reduces excess permanganate) and
aqueous stannous sulfate (reduces ionic mercury to elemental mercury)
were added and the sample was analyzed. The concentration of mercury
was determined by comparing the absorbance of the sample to that
of standards and blanks that were treated in the same manner. The
instrument that was used for this procedure was a Perkin-Elmer 306
atomic absorption spectrophotometer fitted with a cold vapor
mercury analysis apparatus (Olson et a]_., 1975).
14. Total Metals. The sediment sample was placed in the teflon cup of
a Parr acid digestion bomb. A mixture of aqua regia and hydrofluoric
acid was added to the sample and the digestion bomb was sealed and
heated to 120°C for 90 minutes. The digested sample was cooled and
water and boric acid were added to dissolve any metal fluorides
that had formed (Bernas, 1968). The digested samples were analyzed
using a Perkin-Elmer 306 atomic absorption spectrophotometer
equipped with a deuterium arc background corrector and the Perkin-
Elmer HGA 2100 graphite furnace. Samples were analyzed by either
flame or flameless techniques depending on the concentration of
metal in the sample.
B. Water Analysis
The results for chemical analysis of liquid phase samples were listed
in Tables 15 through 26. The methods used in these analyses are summarized
below.
1- Total Sulfide. An unfiltered water sample was preserved by the
addition of zinc acetate. The preserved sample was placed in a
-------
-151-
distillation apparatus, the system was flushed with nitrogen and
the sample was acidified with hydrochloric acid. The sample was
distilled with the distillate collected in 0.2 N zinc acetate
solution. The collected distillate was treated with ferric chloride
and amine-sulfuric acid reagent for color development. The con-
centration of sulfide was determined by comparing the absorbance of
the sample to that of standards that were treated in the same
manner (APHA, 1975).
2. Ammonia Nitrogen. An ammonia electrode (Orion) was used to analyze
the sample. Sodium hydroxide was added to the sample and the amount
of ammonia present was calculated by comparing the sample to various
standards (APHA, 1975).
3. Total Kjeldahl Nitrogen. An unfiltered water sample was treated
with sulfuric acid, potassium sulfate and mercuric sulfate as the
digesting reagent. The digested sample was analyzed using the same
procedure that was previously described for ammonia (APHA, 1975).
4. Organic Nitrogen. This value was calculated as the mathematical
difference between the total Kjeldahl nitrogen and ammonia value
for the same sample.
5. Chemical Oxygen Demand. A water sample was refluxed with a mixture
of potassium dichromate, sulfuric acid, mercuric sulfate and silver
sulfate for 2 hours. The remaining unreacted dichromate was titrated
with standard ferrous ammonium sulfate. The chemical oxygen demand
of the water was calculated by correcting the amount of titrant
used for the sample by the amount used for a blank carried through
the same procedure (APHA, 1975).
6. Orthophosphate. An unfiltered sample was reacted with ammonium
molybdate and stannous chloride. The absorbance of the resulting
blue color was compared to that of standards and blanks treated
-------
-152-
in the same manner (APHA, 1975).
Suspended Solids. The water sample was filtered through a preweighed
0.45 \i membrane filter. The filter was oven-dried at 105°C and
reweighed. The difference in the final and initial weights of the
filter were used to calculate suspended solid values.
Chloride. A titration of the filtered water sample with mercuric
nitrate was performed using diphenylcarbazone as the indicator.
The volume of titrant used for the sample was corrected for the
amount used in titrating a blank (APHA, 1975).
Metal Analysis. The acidified aqueous samples were analyzed
employing a Perkin-Elmer 306 atomic absorption spectrophotometer
equipped with the Perkin-Elmer HGA 2100 graphite furnace and deuterium
arc background correction. The samples were run on either the
flame or furnace attachments depending upon the concentration of
metal in the sample. Concentrations of metal were calculated by
comparing the absorbance of the sample to that standards or by
standard addition if interferences were encountered.
Inorganic Mercury. A digestion of the sample was performed, using
a mixture of nitric acid, sulfuric acid and potassium permanganate
as the digesting reagent. The resulting solution was treated with
a sodium chloride - hydroxylamine sulfate solution and aqueous
stannous sulfate. Liberated mercury was measured on an atomic
absorption spectrophotometer equipped with a cold vapor mercury
apparatus (01 son et_ aj_., 1975).
Total Mercury. The procedure is the same as that used for inorganic
mercury except that potassium persulfate was also added during the
digestion procedure ,(Olson et_ al_., 1975).
Organic Mercury. The organic mercury value was calculated as the
difference between the total and inorganic mercury concentrations.
-------
-153-
13. Dissolved Oxygen. The dissolved oxygen values were determined
using a Yellow Springs Instrument Model 54A Dissolved Oxygen Meter.
The instrument's probe was placed in the sample and moved slowly
until a steady reading was obtained. The instrument was calibrated
by the Winkler titration method (APHA, 1975).
14, Specific Conductance. The measurements were made at 25°C using a
Freas type conductivity cell and an Industrial Instruments Model
RC16B Conductivity Bridge.
-------
-154-
APPENDIX B
1. ANALYSIS OF PCS GAS CHROMATOGRAPHIC ELUTION PATTERNS
2. IDENTIFICATION OF PAHs
-------
1. ANALYSIS OF PCB GAS CHROMATOGRAPHIC ELUTION.PATTERNS _ i$b~-
General Description
The more commonly used commercially available PCB mixtures are desig-
nated as Aroclor 1242, 1248, 1254, 1260 and 1262, A total of 23 different
polychlorinated biphenyl compounds (a number of which are common to more
than one Aroclor mixture) were considered in analyzing samples for these
mixtures. The PCB components present in environmental samples and their
relative abundances do not match any of the commercial mixtures exactly.
However the nature and concentrations of PCB components in an environ-
mental sample may be considered to be a single Aroclor mixture or a linear
combination of two or more mixtures whose individual components have been
altered by solubility differences or degradation by microorganisms.
The computation of PCB concentrations in the environmental samples
were carried out in two ways. The first method involved calculation of
the concentration of each of the possible 23 PCB components commonly
found in samoles (if present) and summing these values to give a total
PCB concentration. The second method consisted of determining the best
fit of the gas chromatographic elution patterns of the five Aroclor
mixtures to the elution pattern of the sample. The concentration of
PCBs in a sample was expressed as specific amounts of one or two Aroclor
mixtures which best matched the sample elution pattern.
Specific Procedure
Total PCBs--
1. The retention times of the peaks from the sample chromatograms were
measured and compared to the retention time of the 23 different PCB peaks
on the same chromatogram. Sample peaks matching the retention times of the
PCB peaks were tentatively identified as PCBs.
^Webb, R.G. and McCal1 , A.C. Quantitative PCB Standards for Electron
Capture Gas Chromatography, J. Chrom. Sci., 11:366-373, 1973.
-------
-156-
2. A computer program was used to calculate the concentration of PCBs
in each sample. Preliminary input information to the computer consisted of:
A. A PCB component response factor for each of the 23 PCB peaks ob-
tained during an earlier series of injections. A response factor
is a quantity representing the mm of elution peak height produced
by the weight of the chromatogramed standard or sample.
B. A conversion factor that adjusts the individual PCB response
factor to the chromatographic conditions of the particular series
of samples being analyzed. This conversion is based on the
relative response factors of the individual PCB components com-
pared to the response factor of a known weight of aldrin under
the same instrumental operating conditions.
3. The specific data that was entered into the computer for calculating
the total concentration of PCBs in a sample is listed below:
A. The injection volume of the standard aldrin solution and the
resulting peak height due to elution of the aldrin.
B. The peak heights of those elution peaks tentatively identified
as PCBs in a given sample.
C. The volume of each sample injected.
D. The initial weight of the sediment sample or initial volume of
the water sample and final volume of the sediment or water extract.
E. The factor by which the extract was diluted.
4. The concentration of each PCB component was comouted using the
following equation:
PCB Cone - (standard conc.) (tl1 standard injected) (peak height in sample)
(pl sample injected) (peak height in standard)
(volume of sample extract)(extract dilution factor)
X
(weight of sediment or volume of water)
-------
157-
5. The concentrations of the individual PCB components were summed
to obtain the total concentration of PCBs.
Aroclor Mixtures—
The computer program contained a procedure designed to analyze the
sample component retention times and relative peak heights by identifying
the Aroclor mixture or mixtures most closely resembling the components
found in the sample. This procedure is summarized as follows.
1. Seven preselected elution bands from each Aroclor standard were
used as references for determining the presence of the commercial
mixture in the sample. The presence and amount of each mixture
was tested in the order 1262, 1260, 1254, 1248 and 1242.
2. The ratio of the peak height of the first elution band (tested band)
to the peak height of the first reference band was computed for the
standard. If the first elution band (test band) and first reference
band were present in the sample, the ratio was also calculated for
the sample. If the ratios for sample and standard matched within
±20%, the concentration of Aroclor 1262 was calculated based on the
peak height of the tested band of the sample. The computation of
ratios of peak heights in the standard and sample based on the first
reference band was repeated for any of the other 21 possible PCB
elution bands which were present in the sample. For those sample
and standard elution band peak height ratios which agreed with ±20%,
concentrations of Aroclor 1262 were computed. All calculated Aroclor
1262 concentration values were averaged and this information was
printed along with the number of PCB tested band-reference band peak
height ratios in the sample that matched elution band-tested
band peak height ratios in the standard.
-------
-158-
3. The procedure described in step 2 was repeated using the other six
reference bands (if present in the sample) chosen in the Aroclor 1262
mixture. The results gave a maximum of six more averaged concentration
values for Aroclor 1262 in the sample. All concentration values of
Aroclor 1262 based on the seven reference elution bands were then
averaged using a weighted average according to the number of sample
and standard peak height ratios which matched within ±20% for each
reference band. This average was reported as the overall average
Aroclor 1262 concentration in the sample.
4. If a larger number of elution bands (generally six or more) from the
sample were used in the calculation of the average Aroclor 1262 con-
centration based on each of the reference elution bands then Aroclor
1262 was assumed to be present in the sample. The peak height contri-
butions due to the presence of the computed concentration of Aroclor
1262 in the sample were subtracted from the observed sample peak heights.
However if only a few elution bands were listed as used in the cal-
culation of the Aroclor 1262 average concentrations, then Aroclor 1262
was assumed to be absent in the sample and this subtraction was not
made.
5. Steps 2 through 4 were repeated for each of the other Aroclor mixtures
in the order 1260, 1254, 1248 and 1242 using the seven selected re-
ference bands for each mixture.
6. After computation of the concentrations of all the Aroclor mixtures
and subtraction of the peak heights due to the presence of the mix-
tures from the observed sample peak heights, the peak heights re-
maining for each of the 23 elution bands were listed.
For the samoles analyzed in this study, it was found that their PCB concen-
trations could be expressed in terms of one or two of the Aroclor mixtures.
-------
-159-
2. IDENTIFICATION OF PAH's
For several samples, benzene eluates from the silica gel fractionation
procedure were chromatographed along with selected PAH standards on temperature-
programmed runs, for comparisons of retention times between peaks in samples
and standards. GC separations were made in a Varian 1700 Series instrument
equipped with a flame-ionization detector, and a glass column (61 x 1/8" ID)
packed with 3% 0V-101 on Gas-Chrom Q. The carrier gas flow rate was 15 ml/min.
A starting column oven temperature of 80 C was held for 1 min, with a programmed
increase of 4 C/min to a final holding temperature of 225 C. Gas chromatograms
of sediment extracts from Lake Superior, site 4, site 6, Pokegama Bay are presented
in Figures B-l to B-4.
Identification of PAH's was performed at the U.S. Environmental Protection
Agency's Environmental Research Laboratory-Duluth on a Finnigan GC-MS system.
A Finnigan Model Model 9610 GC was connected to a Finnigan Model 4032A quadrapole
MS via a glass transfer line. A Finnigan INCOS 2300 data system was used on line
to acquire and process mass spectral data.
A glass GC column (6' x 1/8" ID) was packed with 3% 0V-1 (methyl silicone)
on 60/80 mesh Gas-Chrom Q. The following GC conditions were typically employed:
helium carrier gas flow rate, 20 ml/min; injector temperature, 250 C; separator
oven temperature, 280 C; transfer line temperature, 280 C; and column oven temp-
erature programmed from 100 C to 225 C at 4 C/min.
The mass spectrometer was scanned 50 to 500 at 2.05/decade in electron impact
mode (70 ev). The instrument was tuned to provide a 442/198 amu ratio of 0.7-0.9
for the spectrum of Ultramark 443 (decafluorotriphenylphosphine, PCR Research
Chemicals, Inc.). Calibration was accomplished with FC43 (perfluorinated tributyl
amine).
As an example, Figure B-5 presents a reconstructed ion and selected mass
chromatograms for site 4 sediment extract. Ion intensity, or the number of ion
strikes on the mass spectrometer detector for a compound at a given mass, is
-------
-160-
indicated by the top number of each grouping along the right margin of the
figure. Ion intensity is related to quantity of compound. The mass chromatogram
peaks for the upper lines of the figure are normalized to the compound producing
the greatest ion intensity. In Figure B-5, for example, they are all normalized
to the ion intensity value of 3064 at MW £02. In the left figure margin there
are pairs of numbers associated with the individual mass chromatograms. The lower
number of each pair refers to the MW of the compound producing the mass chromato-
gram, and the upper number refers to percentage ion intensity at a given MW com-
pared to the highest ion intensity of the mass chromatogram displayed.
Mass spectra of unknown compounds from sample extracts were compared to
mass spectra of standard compounds in the EPA/NIH Mass Spectral Data Base (Heller
and Milne, 1978) by a computerized library search. Each search identified three
compounds whose mass spectra best matched mass spectra of unknown compounds.
This listing combined with retention time data of several PAH standards provided
for a positive identification of several PAH's. Examples of library search results
are presented in Figures 6 through 13 for unknown compounds at particular masses
in the sediment extract from site 4.
-------
Figure B-l. FID chromatogram of Lake Superior sediment extract after clean-up by fTori si 1
column chromatography, gel permeation chromatography, and silica gel column chromatography
(benzene eluate).
-------
*srs
Figure B-2. FID chromatogram of Site 4 sediment extract after clean-up by florisil column
chromatography, gel permeation chromatography, and silica gel column chromatography (benzene
eluate). (1) phenanthrene/anthracene, (2) methylphenanthrenes (3) fluoranthene, (4) pyrenes
(5) benz(a)anthracene/triphenylene/chrysene, (6) & (7) elution region of benzo(a)pyrene and
perylene.
-------
6&7
Figure B-3, FID chromatogram of Site 6 sediment extract after clean-up by floris11
column chromatography, gel permeation chromatography, and silica gel column chromatography
(benzene eluate), (1) phenanthrene/anthracene, (2) methylphenanthrene, (3) fluoranthene,
(4) pyrene, (5) benz(a)anthracene/chrysene/triphenylene, (6) & (7) elution region of
benzo(a)pyrene and perylene.
-------
3
Figure B-4. FID chromatogram of Pokegama Bay sediment extract after clean-up by florisil
column chromatography, gel permeation chromatography, and silica gel column Chromatography
(benzene eluate). (1) phenanthrene/anthracene, (2) benz(a)anthracene/chrysene/triphenylene,
(3) unknown.
-------
Figure B-5: Reconstructed Ion and Selected Mass Chromatograms
EIC + MASS CHBDHATOGBAMS DATA: BEEL78224 81
07/31/78 14:16:00 CAti: R73178 #1
SAMPLE: 4 SID
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07/31/78 H:18:60 + 18:24
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-174-
APPENDIX C
CHEMICAL CHARACTERISTICS OF DULUTH-SUPERIOR HARBOR WATER (TABLE C-l),
LAKE SUPERIOR AND CITY OF SUPERIOR WATER (TABLE C-2), AND HEXAGENIA BIOASSAY WATER
AT HIGH D.O. AND LOW D.O. (TABLES C-3, C-4, AND C-5)
-------
-175
CHEMICAL CHARACTERISTICS OF DULUTH-SUPERIOR HARBOR WATER
Measurements of dissolved oxygen (Yellow Springs Model 54A Dissolved
Oxygen Meter), specific conductance and temperature (Yellow Springs Model
33 SCT Meter) were made as a function of depth in the overlying water
at a number of the Duluth-Superior harbor sampling sites in 1977. Similar
measurements were made at the Pokegama Bay reference site. The results
of these measurements are given in Table C-l.
-------
TABLE C-l. TEMPERATURE, DISSOLVED OXYGEN AND SPECIFIC CONDUCTANCE PROFILES OF WATER OVERLYING SITE SEDIMENTS*
SITE DESIGNATION AND WATER CHARACTERISTICS**
June
1
12,
1977
2
June 5,
1977
3
July 10,
1977
4
July 17
, 1977
5A
July 24
, 1977
June
6
26,
1977
Depth(m) Temp.
D.O.
S.C.
Temp.
D.O.
Temp.
D.O.
S.C.
Temp,
D.O.
Temp.
D.O.
Temp,
D.O.
< S.C,
1
15.0
9.4
126
17.0
9.4
18.0
8.5
140
21.2
5.0
23.1
5.9
23.7
5.7
229
2
15.0
9.3
132
15.5
9.4
18.0
8.5
145
21.2
4.7
23.0
5.4
22.0
5.5
222
3
15.0
9.1
_
1 VW
15.0
8.6
17.5
8.3
140
20.8
4.5
21.9
4,6
222
4
15.0
9.0
137
14.5
8.4
17.0
8.2
135
20.2
4.3
21.2
4.7
214
5
1 CO
1 w t L
8.9
138
14.2
8,2
17.0
8.3
130
19.0
3.9
19.5
4.0
210
6
15.2
8.8
138
13.8
8.6
16.8
8.3
120
18.8
4.1
19.0
3.8
208
7
15.3
g "J
139
13.2
9.0
15.9
8.2
115
18 5
1 V*# %
3.9
8
15.3
8.7
140
1 A f
1^.5
9.6
15.0
7.8
120
18.1
3.7
9
16.0
8.5
140
12.0
10.0
17.9
2 8
C— * VJ
11
13
15
* Location of sample sites are described in Section 6,
**Water temperature (Temp.) in degrees Celcius, dissolved oxygen (D.O,) in mg/1 and specific conductance (S.C.)
in micro mhos/cm.
(continued)
-------
Table C-l (continued)
SITE DESIGNATION AND WATER CHARACTERISTICS
LS 6R Pokegama Pokegama Pokegama
August 7, 1977 August 14, 1977 June 6, 1977 July 17, 1977 July 24, 1977
Depth(m) Temp. D.O. S.C. Temp. D.O. Temp. D.O. Temp. D.O. Temp, D.O.
1
18.2
9.7
82
19.0
7 7
2
18.0
83
18.0
7.7
3
17.5
9.9
83
18.0
7.7
4
17.5
—
83
17.5
7,6
5
17.2
10.4
83
17.0
7.1
6
—
—
—
17.0
6,0
7
16.4
10.0
82
8
—
—
—
9
1 C A
ID.4
10.4
81
11
12.9
10.2
78
13
11 .5
10.4
78
15
9.8
— — _
74
-------
-178-
TABLE C-2. COMPARISON OF LAKE SUPERIOR WATER WITH CITY OF SUPERIOR WATER
Dechlorinated City
Lake Superior Water* of Superior Water
Parameter** Mean Range No. Samples Mean+
pH
7.74
7.4-8.2
23
7.1
Total Hardness
45,300
44,000-53,000
54
49,100
A1kalinity
42,300
41,000-50,000
54
49,000
Chloride
1,217
1 ,170-1 ,340
18
4,100
Sodium
1,130
1 ,090-1 ,190
23
o
o
00
Calcium
13,695
13,000-14,700
23
14,600
Magnesium
3,123
2,940-3,590
23
3,300
Potassium
534
480-590
23
520
Iron
23
2-83
10
194
Manganese
0.2-11.5
23
7.8
Chromium
2-20
4
<0.1
Aluminum
—
1-26
5
6
Zinc
0.78
1-2.7
21
3.0
Nickel
<0.5
23
1.3
Lead
—
7-20
2
<0.1
Copper
1.51
0.3-3.2
23
1.3
Cobalt
<0.5
___
23
<0.5
Mercury
<0.01
—
5
—
Cadmium
<0.1
23
<0.0.
* Data from Biesinger and Christensen (1972).
**Values in ug/1 except for pH.
Average of three samples.
-------
TABLE C-3. TEMPERATURE, D.O. AND TURBIDITY CONDITIONS DURING QVERSEDIMENT BIOASSAYS*
Site
Temperature**(°C)
Dissolved 0xygen**(mg/l)
Turbidity"
f"T"l /~\ tr
-------
TABLE C-4. HEXAGENIA POST-BIOASSAY WATER (HIGH D.O.)
SITE
Parameter*
1
2
3
4
5a
6
6R
LS
Pokegema
pH
7.9
7.3
7.1
7.4
7.5
~ —
7,4
7,5
—
Total Phenols
0.015
0.032
0.033
0.027
0.027
0,005
0.042
0.013
0.023
h2s
<0.17
<0.17
0.20
0.20
<0.17
<0.17
<0.17
0.19
<0.17
Ammonia
0.74
2,9
7.5
J Q
2.5
1.7
7.9
5.7
2.6
TKN
4.6
2.9
10.5
16
12
7,2
16
11
7.4
COD
10.3
23.5
17.4
17.0
20.1
5.8
3.9
11,3
21.9
Organic N
3.9
0
3.0
8.1
9.5
5,5
8,1
5,3
4.8
Arsenic
4.1
1.1
3.3
2,6
3.6
3.9
3,9
2.9
3.0
Cadmium
<0.05
<0.05
<0.05
<0,05
<0.05
<0.05
<0,05
<0,05
<0.05
Chromium
1.5
1.1
0.7
0.9
2,2
1.3
1.3
3.6
0.9
Cobalt
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<*n ^
^ U # J
<0,5
A T
<0,5
Copper
13
10
13
10
14
i T
6
10
29
12
Iron
360
570
630
760
630
360
510
510
360
Lead
0.6
0.7
1.4
] 5
1.7
1,0
0,8
A Q
U tO
0.8
Manganese
230
350
115
180
90
130
130
15
27
Nickel
<2.0
<2.0
<2.0
<2,0
<2.0
<2,0
2.1
2.1
<2.0
Selenium
4
1.5
i
1
1
4
1.5
1
4
Zinc
2.2
4.6
<5 5
7 A
5.0
3.2
37
36
4.6
*Values in mg/1
except for
• metals which are given in yg/1.
-------
Parameter*
TABLE
1
C-5.
2
HEXAGENIA POST-BIOASSAY WATER (LOW D.0f)
SITE
3 4 5a 6
6R
LS
Pokegem
pH
7.5
7.1
7.3
6.9
7.3
7 2
7.5
—
Total Phenols
0.014
0.011
0.025
0.034
0,016
0,028
0.016
O.Q2(
h2s
<0.17
<0,17
—
<0.17
<0.17
<0.17
<0J7
0,19
<0.17
Ammonia
1.8
1.7
9.3
8.9
4,6
E E
7,3
5,3
4,3
TKN
3.9
1.7
16
13
13
10
13
12
9
COD
14.0
23.3
18.7
23.5
16.0
6.6
6.3
7.9
2.5
Organic N
2.1
0
6.7
4.1
8.4
4.5
5.7
6.7
4.7
Arsenic
2.6
4.5
3.1
3.6
3.6
5.6
4.6
2.4
2,1
Cadmium
<0.05
<0.05
<0.05
<0.05
<0.05
<0,05
<0.05
0.05
0,05
Chromium
0.5
0.8
0.5
0.6
0.7
4,0
0.8
2.3
1.5
Cobalt
<0.05
<0.05
<0.05
<0.05
<0.05
<0.05
<0,05
<0.05
<0.05
Copper
21
20
9
14
14
10
d
"T
32
17
Iron
210
640
700
760
630
1100
250
510
430
Lead
0.3
0.6
2.1
1.7
1.3
3.2
0.7
1.9
0.8
Manganese
550
610
320
295
110
355
275
19
210
Nickel
<2
<2
<2
<2
<2
< 2
2.1
<2
<2
Selenium
4
4
4
4
i
1
4
4
1
Zinc
1.6
3.2
3.5
10
5.0
5,3
5.0
5.9
4.0
~Values in mg/1
except for metals which are given
in ug/1.
-------
APPENDIX D
GREAT LAKES HARBOR SEDIMENT EVALUATION CRITERIA (EPA)
-------
-183-
GREAT LAKES HARBOR SEDIMENT EVALUATION CRITERIA (EPA)*
Guidelines for the evaluation of Great Lakes harbor sediments, based on
bulk sediment analysis, have been developed by Region V of the U.S. Environ-
mental Protection Agency. These guidelines, developed under the pressure of
the need to make immediate decisions regarding the disposal of dredged material,
have not been adequately related to the impact of the sediments on the lakes
and are considered interim guidelines until more scientifically sound guide-
lines are developed.
The guidelines are based on the following facts and assumptions:
1. Sediments that have been severely altered by the activities of man
are most likely to have adverse environmental impacts.
2. The variability of the sampling and analytical techniques is such
that the assessment of any sample must be based on all factors and
not on any single parameter with the exception of mercury and
polychlorinated biphenyls (PCB's).
3. Due to the documented bioaccumulation of mercury and PCB's, rigid
limitations are used which override all other considerations.
Sediments are classified as heavily polluted, moderately polluted, or
nonpolluted by evaluating each parameter measured against the scales shown
below. The overall classification of the sample is based on the most pre-
dominant classification of the individual parameters. Additional factors
such as elutriate test results, source of contamination, particle size distri-
bution, benthic macroinvertebrate populations, color and odor are also con-
sidered. These factors are interrelated in a complex manner and their inter-
pretation is necessarily somewhat subjective.
*This material is reproduced from the following report:
Guidelines for the Pollution Classification of Great Lakes Harbor Sediments,
U.S. Environmental Protection Agency, Region V, April, 1977,
-------
-184-
Following ranges used to classify sediments from Great Lakes harbors are
based on compilations of data from over 100 different harbors since 1967.
Volatile Solids
COD (mg/kg dry weight)
TKN (mg/kg dry weight)
Oil and Grease
(Hexane Solubles)
(mg/kg dry weight)
Lead (mg/kg dry weight)
Zinc (mg/kg dry weight)
N0NP0LLUTED
<5
<40,000
< 1,000
< 1,000
< 40
< 90
MODERATELY POLLUTED HEAVILY POLLUTED
5-8
40,000-80,000
1 ,000-2,000
1 ,000-2,000
40-60
90-200
>8
>80,000
> 2,000
> 2,000
> 60
> 200
The following supplementary ranges used to classify sediments from Great
Lakes harbors have been developed to the point where they are usable but are
still subject to modification by the addition of new data. These ranges are
based on 260 samples from 34 harbors sampled during 1974 and 1975.
N0NP0LLUTED MODERATELY POLLUTED HEAVILY POLLUTED
Ammonia (mg/kg dry weight)
<75
75-200
>200
Cyanide "
II 14
<0.10
0.10-0.25
>0.25
Phosphorus "
111 U
<420
420-650
>650
Iron
II 11
<17,000
17,000-25,000
>25,000
Nickel "
II II
<20
20-50
>50
Manganese "
II II
<300
300-500
>500
Arsenic "
II II
<3
3-8
>8
Cadmium "
II II
*
*
>6
Chromium "
11 II
<25
25-75
>75
Barium "
11 11
<20
20-60
>60
Copper "
II II
<25
25-50
>50
*Lower limits
not established
-------
-185-
The guidelines stated below for mercury and PCB's are based upon the
best available information and are subject to revision as new information
becomes available.
Methylation of mercury at levels ¦> 1 mg/kg has been documented (1,2).
Methyl mercury is directly available for bioaccumulation in the food chain.
Elevated PCB levels in large fish have been found in all of the Sreat
Lakes. The accumulation pathways are not well understood. However, bio-
accumulation of PCB's at levels >_ 10 mg/kg in fathead minnows has been
documented (3).
Because of the known bioaccumulation of these toxic compounds, a rigid
limitation is used. If the guideline values are exceeded, the sediments are
classified as polluted and unacceptable for open lake disposal no matter what
the other data indicate.
POLLUTED
Mercury >_ 1 mg/kg dry weight
Total PCB's >_ 10 mg/kg dry weight
The pollutional classification of sediments with total PCB concentrations
between 1.0 mg/kg and 10.0 mg/kg dry weight will be determined on a case-by-
case basis.
a. Elutriate test results.
The elutriate test was designed to simulate the dredging and disposal
process. In the test, sediment and dredging site water are mixed in the
ratio of 1:4 by volume. The mixture is shaken for 30 minutes, allowed to
settle for 1 hour, centrifuged, and filtered through a 0.45 P filter. The
filtered water (elutriate water) is then chemically analyzed.
-------
-186-
A sample of the dredging site water used in the elutriate test is
filtered through a 0.45 u filter and chemically analyzed.
A comparison of the elutriate water with the filtered dredging site
water for like constituents indicates whether a constituent was or was
not released in the test.
The value of elutriate test results are limited for overall pollutional
classification because they reflect only immediate release to the water
column under aerobic and near neutral pH conditions. However, elutriate
test results can be used to confirm releases of toxic materials and to in-
fluence decisions where bulk sediment results are marginal between two
classifications. If there is release or non-release, particularly of a
more toxic constituent, the elutriate test results can shift the classifica-
tion toward the more polluted or the less polluted range, respectively.
b. Source of sediment contamination.
In many cases the sources of sediment contamination are readily
apparent. Sediments reflect the inputs of paper mills, steel mills,
sewage discharges and heavy industry very faithfully. Many sediments may
have moderate or high concentrations of TKN, COD, and volatile solids yet
exhibit no evidence of man made pollution. This usually occurs when drain-
age from a swampy area reaches the channel or harbor, or when the project
itself is located in a low lying wetland area. Pollution in these projects
may be considered natural and some leeway may be given in the range values
for TKN, COD, and volatile solids provided that toxic materials are not
also present.
c. Field observations.
Experience has shown that field observations are a most reliable
-------
-187-
indicator of sediment condition. Important factors are color, texture,
odor, presence of detritus, and presence of oily material.
Color. A general guideline is the lighter the color the cleaner
the sediment. There are exceptions to this rule when natural deposits
have a darker color. These conditions are usually apparent to the sedi-
ment sampler during the survey.
Texture. A general rule is the finer the material the more polluted
it is. Sands and gravels usually have low concentrations of pollutants
while silts usually have higher concentrations. Silts are frequently
carried from polluted upstream areas, whereas, sand usually comes from
lateral drift along the shore of the lake. Once again, this general rule
can have exceptions and it must be applied with care.
Odor. This is the odor noted by the sampler when the sample is
collected. These odors can vary widely with temperature and observer and
must be used carefully. Lack of odor, a beach odor, or a fishy odor tends
to denote cleaner samples.
Detritus. Detritus may cause higher values for the organic parameters
COD, TKN, and volatile solids. It usually denotes pollution from natural
sources. Note: The determination of the "naturalness" of a sediment depends
upon the establishment of a natural organic source and a lack of man made
pollution sources with low values for metals and oil and grease. The pres-
ence of detritus is not decisive in itself.
Oily material. This almost always comes from industry or shipping
activities. Samples showing visible oil are usually highly contaminated.
If chemical results are marginal, a notation of oil is grounds for declaring
the sediment to be polluted.
-------
-188-
d. Benthos,
Classical biological evaluation of benthos is not applicable to harbor
or channel sediments because these areas very seldom support a well balanced
population. Very high concentrations of tolerant organisms indicate organic
contamination but do not necessarily preclude open lake disposal of the
sediments. A moderate concentration of oligochaetes or other tolerant
organisms frequently characterizes an acceptable sample. The worst case
exists when there is a complete lack or very limited number of organisms.
This may indicate a toxic condition.
In addition, biological results must be interpreted in light of the
habitat provided in the harbor or channel. Drifting sand can be a very
harsh habitat which may support only a few organisms. Silty materials on
the other hand, usually provides a good habitat for sludgeworms, leeches,
fingernail clams, and perhaps, amphipods. Material that is frequently
disturbed by ship's propellers provides a poor habitat.
-------
189-
REFERENCES
1. Jensen, S., and Jernelov, A., "Biological Methylation of Mercury In
Aquatic OrganismsNature, 223, August 16, 1969 pp 753-754.
2. Magnuson, J.J., Forbes, A., and Hall, R., "Final Report - An Assessment
of the Environmental Effects of Dredged Material Disposal in Lake
Superior - Volume 3: Biological Studies," Marine Studies Center,
University of Wisconsin, Madison, March, 1976.
3. Halter, M.T., and Johnson, H.E., "A Model System to Study the Release
of PCB from HydrosoiIs and Subsequent Accumulation by Fish,"
presented to American Society for Testing and Materials, Symposium
on Aquatic Toxicology and Hazard Evaluation," October 25-26, 1976,
Memphis, Tennessee.
-------
TECHNICAL REPORT DATA
(Please read Instructions on the reverse before completing)
1. REPORT NO. 2.
EPA-600/3-81-02S
3. RECIPIENT'S ACCESSION NO.
^ < !*S26 1
4. TITLE AND SUBTITLE
Development of Bioassay Procedures for Defining
Pollution of Harbor Sediments Part I
5, REPORT DATE
MARCH 1981 ISSUING DATE.
8, PERFORMING ORGANIZATION CODE
7. AUTHOR(S)
Donald A, Bahnick, William A. Swenson, Thomas P. Markee,
Daniel J. Call, Craig A, Anderson, and R. Ted Morris
8. PERFORMING ORGANIZATION REPORT NO.
9. PERFORMING ORGANIZATION NAME AND ADDRESS
Center for Lake Superior Environmental Studies
University of Wisconsin-Superior
Superior5 Wisconsin
10. PROGRAM ELEMENT NO.
11. CONTRACT/GRANT NO.
R804918-01
12. SPONSORING AGENCY NAME AND ADDRESS
U.S. Environmental Protection Agency
Environmental Research Laboratory-Duluth
6201 Congdon Boulevard
Duluth, Minnesota 55804
13, TYPE OF REPORT AND PERIOD COVERED
14. SPONSORING AGENCY CODE
EPA-600/03
15. SUPPLEMENTARY NOTES
16. ABSTRACT
This research investigates bioassay methods which may be useful in assessing the
degree of pollution of harbor sediments. Procedures studied include 96 hr toxicity
tests employing Hexagenia limbata, Daphnia magna and Pontoporeia affinis as biological
probes. monitoring cough frequencies of bluegill sunfish (Lepomis taacrochirus) in
interstitial water derived from sediments, chemical analyses of sediment-water systems,
and chemical analysis of chironomids and Hexagenia limbata exposed to the sediments.
Additional experiments involved investigation of the degree of removal of chemical
constituents from sediments due to extraction with Lake Superior water and the use of
reverse phase liquid chromatography in detecting the presence of chemical compounds
with high bioaccumulation potential in the sediments.
A general toxicity index was prepared from the chemical data which indicated that
animal survival in the 96 hr acute toxicity tests was generally lower using sediment
systems from the more industrialized areas of the harbor.
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EPA Form 2220-1 (Rev, 4-77) previous edition is obsolete
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