PEER REVIEW DRAFT, DO NOT CITE OR QUOTE
&EPA
United States
Environmental Protection Agency
Office of Chemical Safety and
Pollution Prevention
Risk Evaluation for
Methylene Chloride
Systematic Review Supplemental File:
Data Quality Evaluation of Environmental Hazard Studies
CASRN: 75-09-2
H
H

CI
October 2019, DRAFT

-------
1
4
7
9
11
13
16
19
22
24
Table of Contents
Data Type
Acute (0-96 hour); Aquatic; Inver-
tebrates
Acute (0-96 hour); Aquatic; Fish
Acute (0-96 hour); Aquatic; Fish
Other; Aquatic; Invertebrates
Acute (0-96 hour); Aquatic; Inver-
tebrates
Acute (0-96 hour); Aquatic; Fish
Acute (0-96 hour); Aquatic; other
Fish; Static Test
Acute (0-96 hour); Aquatic; other
Fish; Flow-Through Test
Acute (0-96 hour); Aquatic; Inver-
tebrates
Acute (0-96 hour); Aquatic; other
Amphibians
Reference
Leblanc, G. A.. 1980. Acute toxicity of priority pollutants to water flea (Daph-
nia magna). Bulletin of Environmental Contamination and Toxicology 24:684-
691
Buccafusco, R. J.,Ells, S. J.,Leblanc, G. A.. 1981. Acute toxicity of priority
pollutants to bluegill (Lepomis macrochirus). Bulletin of Environmental Con-
tamination and Toxicology 26:446-452
Heitmuller, P. T.,Hollister, T. A.,Parrish, P. R.. 1981. Acute toxicity of 54
industrial chemicals to sheepshead minnows (Cyprinodon variegatus). Bulletin
of Environmental Contamination and Toxicology 27:596-604
Samoiloff, M.R., Schulz, S., Jordan, Y., Denich, K., Arnott, E.. 1980. A rapid
simple long-term toxicity assay for aquatic contaminants using the nematode
Panagrellus redivivus. Can. J. Fish. Aquat. Sci. 37:1167-1174
Samoiloff, M.R., Schulz, S., Jordan, Y., Denich, K., Arnott, E.. 1980. A rapid
simple long-term toxicity assay for aquatic contaminants using the nematode
Panagrellus redivivus. Can. J. Fish. Aquat. Sci. 37:1167-1174
Geiger, D. L.,Poirier, S. H.,Brooke, L. T.,Call, D. J. eds. 1986. Acute toxicities
of organic chemicals to fathead minnows (Pimephales promelas): volume III.
Alexander, H. C.,McCarty, W. M.,Bartlett, E. A.. 1978. Toxicity of per-
chloroethylene, trichloroethylene, 1,1,1-trichloroethane, and methylene chloride
to fathead minnows. Bulletin of Environmental Contamination and Toxicology
20:344-352
Alexander, H. C.,McCarty, W. M.,Bartlett, E. A.. 1978. Toxicity of per-
chloroethylene, trichloroethylene, 1,1,1-trichloroethane, and methylene chloride
to fathead minnows. Bulletin of Environmental Contamination and Toxicology
20:344-352
Kuhn, R.,Pattard, M.,Pernak, K. D.,Winter, A.. 1989. Results of the harmful
effects of selected water pollutants (anilines, phenols, aliphatic compounds) to
Daphnia magna. Water Research 23:495-499
Black, J. A.,Birge, W. J.,McDonnell, W. E.,Westerman, A. G.,Ramey, B.
A.,Bruser, D. M.. 1982. The aquatic toxicity of organic compounds to embryo-
larval stages of fish and amphibians. 133
l.

-------
27
30
33
36
38
41
44
47
49
52
55
58
Acute (0-96 hour); Aquatic; Fish
Acute (0-96 hour); Aquatic; Inver-
tebrates
Acute (0-96 hour); Aquatic; Plants
Acute (0-96 hour); Aquatic; other
Amphibians
Acute (0-96 hour); Aquatic; Inver-
tebrates
Acute (0-96 hour); Aquatic; other
fathead minnow cell-line; total pro-
tein content
Acute (0-96 hour); Aquatic; other
Algae; MDA
Acute (0-96 hour); Aquatic; other
Algae; Chlorophyl a
Acute (0-96 hour); Aquatic; other
Algae; Protein Content
Acute (0-96 hour); Aquatic; other
Algae; CAT and SOD
Acute (0-96 hour); Aquatic; other
Algae; Growth
Acute (0-96 hour); Aquatic; other
soil fungi
Black, J. A.,Birge, W. J.,McDonnell, W. E.,Westerman, A. G.,Ramey, B.
A.,Bruser, D. M.. 1982. The aquatic toxicity of organic compounds to embryo-
larval stages of fish and amphibians. 133
Sanchez-Fortun, S.,Sanz, F.,Santa-Maria, A.,Ros, J. M.,De Vicente, M.
L.,Encinas, M. T.,Vinagre, E.,Barahona, M. V.. 1997. Acute sensitivity of
three age classes of Artemia salina larvae to seven chlorinated solvents. Bul-
letin of Environmental Contamination and Toxicology 59:445-451
Brack, W.,Rottler, H.. 1994. Toxicity testing of highly volatile chemicals with
green algae: A new assay. 1:223-228
Marquis, 0.,Millery, A.,Guittonneau, S.,Miaud, C.. 2006. Solvent toxicity to
amphibian embryos and larvae. Chemosphere 63:889-892
Abernethy, S.,Bobra, A. M.,Shiu, W. Y.,Wells, P. G.,Mackay, D.. 1986. ACUTE
LETHAL TOXICITY OF HYDROCARBONS AND CHLORINATED HY-
DROCARBONS TO TWO PLANKTONIC CRUSTACEANS THE KEY ROLE
OF ORGANISM-WATER PARTITIONING. Aquatic Toxicology
Dierickx, P. J.. 1993. Comparison between fish lethality data and the in vitro
cytotoxicity of lipophilic solvents to cultured fish cells in a two-compartment
model. Chemosphere 27:1511-1518
Wu, S.,Zhang, H.,Yu, X.,Qiu, L.. 2014. Toxicological Responses of Chlorella
vulgaris to Dichloromethane and Dichloroethane. Environmental Engineering
Science 31
Wu, S.,Zhang, H.,Yu, X.,Qiu, L.. 2014. Toxicological Responses of Chlorella
vulgaris to Dichloromethane and Dichloroethane. Environmental Engineering
Science 31
Wu, S.,Zhang, H.,Yu, X.,Qiu, L.. 2014. Toxicological Responses of Chlorella
vulgaris to Dichloromethane and Dichloroethane. Environmental Engineering
Science 31
Wu, S.,Zhang, H.,Yu, X.,Qiu, L.. 2014. Toxicological Responses of Chlorella
vulgaris to Dichloromethane and Dichloroethane. Environmental Engineering
Science 31
Wu, S.,Zhang, H.,Yu, X.,Qiu, L.. 2014. Toxicological Responses of Chlorella
vulgaris to Dichloromethane and Dichloroethane. Environmental Engineering
Science 31
Steiman, R.,Seiglemurandi, F.,Guiraud, P.,Benoitguyod, J. L.. 1995. TESTING
OF CHLORINATED SOLVENTS ON MICROFUNGI. Environmental Toxicol-
ogy and Water Quality 10:283-285

-------
60
63
65
68
71
74
77
79
82
85
87
90
Other; Aquatic; Fish
Other; Aquatic; other mesocosm
BCF
Chronic (>21 days); Aquatic; In-
vertebrates
Acute (0-96 hour); Aquatic; Inver-
tebrates
Acute (0-96 hour); Aquatic; other
Amphibians
Other; Aquatic; other Algae;
Chlorophyll a
Aquatic; Invertebrates
Acute (0-96 hour); Aquatic; Plants
Acute (0-96 hour); Aquatic; other
Insect-larvae 4-hour LC50
Acute (0-96 hour); Aquatic; other
Mixture 63 percent DCM; fathead
minnow
Acute (0-96 hour); Aquatic; Fish;
rainbow trout
Dill, D. C., Murphy, P. G.,Mayes, M. A.. 1987. TOXICITY OF
METHYLENE-CHLORIDE TO LIFE STAGES OF THE FATHEAD MIN-
NOW, PIMEPHALES-PROMELAS RAFINESQUE. Bulletin of Environmental
Contamination and Toxicology 39:869-876
Thiebaud, H.,Merlin, G.,Capovilla, M. P.,Blake, G.. 1994. Fate of a volatile
chlorinated solvent in indoor aquatic microcosms: sublethal and static exposure
to [14C]dichloromethane. Groupe pour l'Etude du Devenir de X" nobiotiques
dans PEnvironnement (GEDEXE). Ecotoxicology and Environmental Safety
28:71-81
Rayburn, J. R.,Fisher, W. S.. 1999. Developmental toxicity of copper chloride,
methylene chloride, and 6-aminonicotinamide to embryos of the grass shrimp
Palaemonetes pugio. Environmental Toxicology and Chemistry 18:950-957
Rayburn, J. R.,Fisher, W. S.. 1999. Developmental toxicity of copper chloride,
methylene chloride, and 6-aminonicotinamide to embryos of the grass shrimp
Palaemonetes pugio. Environmental Toxicology and Chemistry 18:950-957
Birge, W. J.,Black, J. A.,Kuehne, R. A.. 1980. Eifects of Organic Compounds on
Amphibian Reproduction.
Ando, T.,Otsuka, S.,Nishiyama, M.,Senoo, K.,Watanabe, M. M.,Matsumoto, S..
2003. Toxic Eifects of Dichloromethane and Trichloroethylene on the Growth of
Planktonic Green Algae, Chlorella vulgaris NIES227, Selenastrum capricor-
nutum NIES35, and Volvulina steinii NIES545. 18:43-46
Wilson, J. E. H.. 1998. Developmental Arrest in Grass Shrimp Embryos Ex-
posed to Selected Toxicants.
Tsai, K. P.,Chen, C. Y.. 2007. An Algal Toxicity Database of Organic Toxi-
cants Derived by a Closed-System Technique. Environmental Toxicology and
Chemistry 26:1931-1939
Kramer, V. C.,Schnell, D. J.,Nickerson, K. W.. 1983. Relative Toxicity of
Organic Solvents to Aedes aegypti Larvae. 42:285-287
Minnesota Mining & Mfg Co. 1979. 96-HOUR LC50 AQUATIC TEST ON
FATHEAD MINNOWS WITH COVER LETTER.
E I Dupont Denemours & Co Inc. 1987. FLOW-THROUGH ACUTE 96-HOUR
LC50 OF METHYLENE CHLORIDE TO RAINBOW TROUT (SANITIZED).
Acute (0-96 hour); Aquatic; Inver-
tebrates
E I Dupont Denemours & Co Inc. 1987. DAPHNIA MAGNA STATIC ACUTE
48-HOUR EC50 OF METHYLENE CHLORIDE (SANITIZED).

-------
Study Citation: Leblanc, G. A.. 1980. Acute toxicity of priority pollutants to water flea (Daphnia magna). Bulletin of Environmental Contamination
and Toxicology 24:684-691
Data Type:	Acute (0-96 hour); Aquatic; Invertebrates
Hero ID:	7508
Domain
Metric
Rating^
MWF*
Score
Comments it
Domain 1: Test Substance






Metric 1:
Test Substance Identity
High
X
2
2
Test substance was identified as dichloromethane
Metric 2:
Test Substance Source
Medium
X
1
2
Indicated from commercial sources, but source never
identified.
Metric 3:
Test Substance Purity
Low
X
1
3
Indicated > = 80 percent purity for all chemicals
tested; but not specific for dichlomethane.
Domain 2: Test Design






Metric 4:
Negative Controls
High
X
2
2
A diluent water negative control was used.
Metric 5:
Negative Control Response
High
X
1
1
Less than 10 percent morality in control populations.
Metric 6:
Randomized Allocation
Medium
X
1
2
Random allocation indicated, but method used for
randomization is unclear.
Domain 3: Exposure Characterization





Metric 7:
Experimental System/Test Media Prepara-
Low
X
2
6
Indicated 15 daphnids in one test vessel covered with

tion




plastic wrap to reduce risk of loss of substance via





volatilization, but did not reduce headspace so some
volatilization expected.
Metric 8:
Consistency of Exposure Administration
High
X
1
1
Distilled water was used to prepare stock solutions
of water soluble test substances.
Metric 9:
Measurement of Test Substance Concentra-
Low
X
1
3
Nominal concentrations, attempt to reduce loss of

tion




substance via volatilization but uncertain whether





sufficient measures to reduce loss.
Metric 10:
Exposure Duration and Frequency
High
X
2
2
Test duration of 48-hours was adequate for deter-
mining effects on Daphnia magna.
Metric 11:
Number of Exposure Groups/Spacing of Ex-
Medium
X
1
2
Indicated 5-8 nominal concentrations, but number

posure Levels




of concentrations specific for methylene chloride and





spacing of test concentrations is unclear.
Metric 12:
Testing at or Below Solubility Limit
High
X
1
1
Test results are below the water solubility of methy-
lene chloride.
Domain 4: Test Organism
Continued on next page . . .
1

-------
. . continued from previous page
Study Citation: Leblanc, G. A.. 1980. Acute toxicity of priority pollutants to water flea (Daphnia magna). Bulletin of Environmental Contamination
and Toxicology 24:684-691
Data Type:	Acute (0-96 hour); Aquatic; Invertebrates
Hero ID:	7508
Domain
Metric
Rating"!" MWF* Score
Comments'^
Metric 13: Test Organism Characteristics	High x 2 2
Metric 14: Acclimitization and Pretreatment Conditions Medium x 1	2
Metric 15: Number of Organisms and Replicates per High x 1	1
Group
Metric 16: Adequacy of Test Conditions	Medium x 1	2
Daphnia magna are well-known aquatic inverte-
brates and suitable for freshwater toxicity tests.
Did not indicate pretreatment conditions, but re-
ferred to use of procedures from "methods for acute
toxicity tests with fish, macroinvertebrates and am-
phibians (U.S. EPA 1975).
There were 15 daphnia per test concentration and 3
replicates tested.
Referred to procedures from "Methods for acute tox-
icity tests with fish, macroinvertebrates and amphib-
ians (U.S. EPA 1975)."
Domain 5: Outcome Assessment
Metric 17: Outcome Assessment Methodology
Metric 18: Consistency of Outcome Assessment
High
High
x 2
x 1
Daphnia mortality was recorded at 24 hours and 48
hours, and water quality characteristics (pH, dis-
solved oxygen, and temperature) were measured at
test initiation and termination.
Domain 6: Confounding / Variable Control
Metric 19: Confounding Variables in Test Design and High x 2
Procedures
Metric 20: Outcomes Unrelated to Exposure
High
x 1
Water quality parameters appeared to be within ac-
ceptable limits for daphnia magna as characterized
by OECD Test Guideline 202.
Domain 7: Data Presentation and Analysis
Metric 21: Statistical Methods
Medium x 1
Metric 22: Reporting of Data
Metric 23: Explanation of Unexpected Outcomes
High
High
x 2
x 1
Author appeared to use the moving average angle
method with 95 percent confidence limits, based on
nominal concentrations, to calculate the LC50 for
methylene chloride, but the data to reproduce these
result were not provided.
Control mortality was less than 10 percent.
Overall Quality Determination"'"	High	1.5
Continued on next page . . .
2

-------
. . . continued from previous page
Study Citation: Leblanc, G. A.. 1980. Acute toxicity of priority pollutants to water flea (Daphnia magna). Bulletin of Environmental Contamination
and Toxicology 24:684-691
Data Type:	Acute (0-96 hour); Aquatic; Invertebrates
Hero ID:	7508
Domain
Metric
Rating"!" MWF* Score
Comments'^
Extracted
Yes
* MWF = Metric Weighting Factor
"!" High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value.
^ The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High.
Overall rating =
(Metric Score; X MWF;) / J] . MWF,
if any metric is Unacceptable
(round to the nearest tenth) otherwise
where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is
crossed out and an arrow points to the new rating.
Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments.
3

-------
Study Citation: Buccafusco, R. J.,Ells, S. J.,Leblanc, G. A.. 1981. Acute toxicity of priority pollutants to bluegill (Lepomis macrochirus). Bulletin of
Environmental Contamination and Toxicology 26:446-452
Data Type:	Acute (0-96 hour); Aquatic; Fish
Hero ID:	18064
Domain
Metric
Rating1
MWF* Score
Comments' t
Domain 1: Test Substance
Metric 1:
Metric 2:
Test Substance Identity
Test Substance Source
Metric 3: Test Substance Purity
High	X 2	2 Test substance identified as dichloromethane.
Low	X 1	3 The study indicates that all chemicals tested were
purchased from commercial chemical suppliers, but
does not specify what company.
Medium	X 1	2 Study reports a minimum purity of 80 percent for
all chemicals tested, but does not specify what the
purity is for DCM.
Domain 2: Test Design
Metric 4:
Metric 5:
Negative Controls
Negative Control Response
Metric 6: Randomized Allocation
High
Low
High
x 2
x 1
x 1
Control mortality is mentioned, but no further de-
tails.
Domain 3: Exposure Characterization
Metric 7:
Experimental System/Test Media Prepara- Unacceptable x 2
tion
Metric 8: Consistency of Exposure Administration	High	x 1
Metric 9: Measurement of Test Substance Concentra- Unacceptable x 1
tion
Metric 10: Exposure Duration and Frequency	High
Metric 11: Number of Exposure Groups/Spacing of Ex- Low
posure Levels
Metric 12: Testing at or Below Solubility Limit
Low
x 2
x 1
x 1
Did not cap DCM as with other volatile chemicals
tested.
Nominal concentrations were used and were not
measured. DCM is volatile.
Study used EPA's "Methods for acute toxicity
tests with fish, macroinvertebrates, and amphib-
ians" which requires that static tests have 10 organ-
isms/treatment divided into >= two test chambers.
Test concentrations and spacing were not specified.
Test substance concentration was not reported, and
it was reported that for some concentrations there
was undissolved chemical.
Domain 4: Test Organism
Continued on next page
4

-------
. . . continued from previous page
Study Citation: Buccafusco, R. J.,Ells, S. J.,Leblanc, G. A.. 1981. Acute toxicity of priority pollutants to bluegill (Lepomis macrochirus). Bulletin of
Environmental Contamination and Toxicology 26:446-452
Data Type:	Acute (0-96 hour); Aquatic; Fish
Hero ID:	18064
Domain
Metric
Rating1
MWF* Score
Comments' t
Metric 13: Test Organism Characteristics
Medium
Metric 14: Acclimitization and Pretreatment Conditions	Medium
Metric 15: Number of Organisms and Replicates per	Medium
Group
Metric 16: Adequacy of Test Conditions	Medium
x 2
x 1
x 1
x 1
Test animals utilized were young of the year bluegill
(L. macrochirus) obtained from commercial fish sup-
pliers within the continental United States.
Fish were observed for 48 hours and not used if had
>3 percent mortality, but acclimation time (i.e. 12
days in the lab) was not reported.
There are minor uncertainties around number of or-
ganisms used.
There are minor uncertainties around housing con-
ditions (ex. headspace in jar, DO cones).
Domain 5: Outcome Assessment
Metric 17: Outcome Assessment Methodology
Metric 18: Consistency of Outcome Assessment
High
High
x 2
x 1
Domain 6: Confounding / Variable Control
Metric 19: Confounding Variables in Test Design and Low
Procedures
Metric 20: Outcomes Unrelated to Exposure	Low
X 2	6	The study did not provide enough information to
allow a comparison of environmental conditions
X 1	3	The study did not provide enough information about
health outcomes of each study group.
Domain 7: Data Presentation and Analysis
Metric 21: Statistical Methods
Metric 22: Reporting of Data
Metric 23: Explanation of Unexpected Outcomes
Medium
Low
High
x 1
x 2
x 1
The method used to calculate LC50s for DCM is un-
clear: Harris method (Harris, 1959) or the log probit
method, a modification of the Litchfield and Wilcox
(1949) method.
Exposure-related behavioral effects not reported,
only mortality, and effects at each test concentration
(e.g. percent mortality at lowest through highest
concentration tested) not provided, including con-
trols.
Overall Quality Determination"'"
Unacceptable
4.0
Met
ric mean score
2.1.
Continued on next page
5

-------
. continued from previous page
Study Citation: Buccafusco, R. J.,Ells, S. J.,Leblanc, G. A.. 1981. Acute toxicity of priority pollutants to bluegill (Lepomis macrochirus). Bulletin of
Environmental Contamination and Toxicology 26:446-452
Data Type:	Acute (0-96 hour); Aquatic; Fish
Hero ID:	18064
Domain
Metric
Rating1
MWF* Score
Comments' t
Extracted

No



** Consistent with our Application of Systematic Review in TSCARisk Evaluations document, if a metric for a data source receives a score of Unacceptable (score = 4),
EPA will determine the study to be unacceptable. In this case, two of the metrics were rated as unacceptable. As such, the study is considered unacceptable and the score
is presented solely to increase transparency.
*	MWF = Metric Weighting Factor
r High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value.
*	The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High.
Overall rating =
(Metric Score; x MWF;) / J] . MWFj
if any metric is Unacceptable
(round to the nearest tenth) otherwise
where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed
out and an arrow points to the new rating,
ft Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments.
6

-------
Study Citation: Heitmuller, P. T.,Hollister, T. A.,Parrish, P. R.. 1981. Acute toxicity of 54 industrial chemicals to sheepshead minnows (Cyprinodon
variegatus). Bulletin of Environmental Contamination and Toxicology 27:596-604
Data Type:	Acute (0-96 hour); Aquatic; Fish
Hero ID:	18110
Domain
Metric
Rating1
MWF*
Score
Comments' t
Domain 1: Test Substance






Metric 1:
Test Substance Identity
High
X
2
2

Metric 2:
Test Substance Source
Medium
X
1
2
Unspecified commercial source.
Metric 3:
Test Substance Purity
Medium
X
1
2
>=80 percent purity, but not specified for DCM.
Domain 2: Test Design






Metric 4:
Negative Controls
High
X
2
2

Metric 5:
Negative Control Response
Medium
X
1
2
Indicated test not acceptable if control mortality ex-






ceeded 10 percent but no specific details for DCM
Metric 6:
Randomized Allocation
Low
X
1
3
Randomized allocation not indicated.
Domain 3: Exposure Characterization





Metric 7:
Experimental System/Test Media Prepara-
Unacceptable
X
2
8
Static system, did not take measures to control

tion




volatilization of methylene chloride.
Metric 8:
Consistency of Exposure Administration
High
X
1
1
Exposures consistent across study groups.
Metric 9:
Measurement of Test Substance Concentra-
Unacceptable
X
1
4
No analytical monitoring; Nominal concentrations

tion




used and methylene chloride is volatile.
Metric 10:
Exposure Duration and Frequency
High
X
2
2

Metric 11:
Number of Exposure Groups/Spacing of Ex-
Low
X
1
3
Test concentrations determined after range-finding

posure Levels




test were not specified.
Metric 12:
Testing at or Below Solubility Limit
Low
X
1
3
Not specified so uncertain.
Domain 4: Test Organism






Metric 13:
Test Organism Characteristics
High
X
2
2

Metric 14:
Acclimitization and Pretreatment Conditions
Medium
X
1
2
Total holding period not indicated.
Metric 15:
Number of Organisms and Replicates per
High
X
1
1


Group





Metric 16:
Adequacy of Test Conditions
High
X
1
1
housing prior to testing not clear but flowing seawa-
ter and feeding conditions indicated.
Continued on next page . . .
7

-------
. continued from previous page
Study Citation: Heitmuller, P. T.,Hollister, T. A.,Parrish, P. R.. 1981. Acute toxicity of 54 industrial chemicals to sheepshead minnows (Cyprinodon
variegatus). Bulletin of Environmental Contamination and Toxicology 27:596-604
Data Type:	Acute (0-96 hour); Aquatic; Fish
Hero ID:	18110
Domain Metric
Rating1
MWF*
Score
Comments' t
Domain 5: Outcome Assessment





Metric 17: Outcome Assessment Methodology
High
X
2
2

Metric 18: Consistency of Outcome Assessment
High
X
1
1

Domain 6: Confounding / Variable Control





Metric 19: Confounding Variables in Test Design and
Medium
X
2
4
Uncertain due to minimal details provided for test
Procedures




setup and test methods for this multichemical test-




ing study.
Metric 20: Outcomes Unrelated to Exposure
Medium
X
1
2
Minimal details provided - no health outcomes not





related to exposures indicated, but assumed that





multichemical test will report significant health is-





sues/mortality.
Domain 7: Data Presentation and Analysis





Metric 21: Statistical Methods
Medium
X
1
2
Moving average angle analysis, probit analysis, or





binomial probability used, but not clear which for





DCM.
Metric 22: Reporting of Data
Medium
X
2
4
Exposure-related behavioral effects not reported,





only mortality, and effects at each test concentra-





tion (e.g. percent mortality at lowest through high-





est concentration tested) not provided.
Metric 23: Explanation of Unexpected Outcomes
High
X
1
1

Overall Quality Determination"'"
Unacceptable


4.0
Metric mean score**: 1.8.
Extracted
No
** Consistent with our Application of Systematic Review in TSCARisk Evaluations document, if a metric for a data source receives a score of Unacceptable (score = 4),
EPA will determine the study to be unacceptable. In this case, two of the metrics were rated as unacceptable. As such, the study is considered unacceptable and the score
is presented solely to increase transparency.
*	MWF = Metric Weighting Factor
t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value.
*	The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High.
Overall rating =
(Metric Score; x MWF;) / ^ MWFj
if any metric is Unacceptable
(round to the nearest tenth) otherwise
where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed
out and an arrow points to the new rating.
^ Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments.
8

-------
Study Citation: Samoiloff, M.R., Schulz, S., Jordan, Y., Denich, K., Arnott, E.. 1980. A rapid simple long-term toxicity assay for aquatic contaminants
using the nematode Panagrellus redivivus. Can. J. Fish. Aquat. Sci. 37:1167-1174
Data Type:	Other; Aquatic; Invertebrates
Hero ID:	29147
Domain
Metric
Rating1
MWF* Score
Comments' t
Domain 1: Test Substance
Metric 1:
Metric 2:
Test Substance Identity
Test Substance Source
Metric 3: Test Substance Purity
High
Medium
Low
x 2
x 1
x 1
Two potential source companies indicated, but not
specific for methylene chloride.
Source company indicated, suggests at least a tech-
nical grade; but no purity given.
Domain 2: Test Design
Metric 4:
Metric 5:
Metric 6:
Negative Controls
Negative Control Response
Randomized Allocation
High
Low
Low
x 2
x 1
x 1
Negative control responses not provided.
Randomized allocation not indicated.
Domain 3: Exposure Characterization
Metric 7: Experimental System/Test Media Prepara-
tion
Metric 8: Consistency of Exposure Administration
Metric 9: Measurement of Test Substance Concentra-
tion
Metric 10: Exposure Duration and Frequency
Metric 11: Number of Exposure Groups/Spacing of Ex-
posure Levels
Metric 12: Testing at or Below Solubility Limit
Low
High
N/A
High
High
High
x 2
x 1
x 2
x 1
x 1
6
1
N/A
DCM's volatility was not addressed.
Testing conducted in a medium that would interfere
with analytical instruments. Effects were observed
at low concentrations suggesting DCM was main-
tained in the medium.
Domain 4: Test Organism
Metric 13: Test Organism Characteristics
Unacceptable x 2
Metric 14: Acclimitization and Pretreatment Conditions High
Metric 15: Number of Organisms and Replicates per High
Group
x 1
x 1
The nematode is not a well-known laboratory
species, and is not representative of aquatic inver-
tebrates.
Continued on next page . . .
9

-------
. .. continued from previous page
Study Citation: Samoiloff, M.R., Schulz, S., Jordan, Y., Denich, K., Arnott, E.. 1980. A rapid simple long-term toxicity assay for aquatic contaminants
using the nematode Panagrellus redivivus. Can. J. Fish. Aquat. Sci. 37:1167-1174
Data Type:	Other; Aquatic; Invertebrates
Hero ID:	29147
Domain Metric
Rating1
MWF*
Score
Comments' t
Metric 16: Adequacy of Test Conditions
Medium
X
1
2
No details on biomass loading, temperature, etc were





provided.
Domain 5: Outcome Assessment





Metric 17: Outcome Assessment Methodology
High
X
2
2

Metric 18: Consistency of Outcome Assessment
High
X
1
1

Domain 6: Confounding / Variable Control





Metric 19: Confounding Variables in Test Design and
Low
X
2
6
Study did not provide information on individual en-
Procedures




vironmental conditions for each chemical tested to




allow a comparison.
Metric 20: Outcomes Unrelated to Exposure
Low
X
1
3
Not indicated.
Domain 7: Data Presentation and Analysis





Metric 21: Statistical Methods
High
X
1
1

Metric 22: Reporting of Data
Medium
X
2
4
Control results not provided.
Metric 23: Explanation of Unexpected Outcomes
High
X
1
1

Overall Quality Determination"'"
Unacceptable


4.0
Metric mean score**: 1.9.
Extracted
No




** Consistent with our Application of Systematic Review in TSCARisk Evaluations document, if a metric for a data source receives a score of Unacceptable (score = 4),
EPA will determine the study to be unacceptable. In this case, one of the metrics were rated as unacceptable. As such, the study is considered unacceptable and the score
is presented solely to increase transparency.
*	MWF = Metric Weighting Factor
t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value.
*	The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High.
Overall rating =
^ (Metric Score; x MWF;) / ^ MWFj
if any metric is Unacceptable
(round to the nearest tenth) otherwise
where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed
out and an arrow points to the new rating.
^ Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments.
10

-------
Study Citation: Samoiloff, M.R., Schulz, S., Jordan, Y., Denich, K., Arnott, E.. 1980. A rapid simple long-term toxicity assay for aquatic contaminants
using the nematode Panagrellus redivivus. Can. J. Fish. Aquat. Sci. 37:1167-1174
Data Type:	Acute (0-96 hour); Aquatic; Invertebrates
Hero ID:	29147
Domain	Metric	Rating^ MWF* Score	Comments^
Domain 1: Test Substance
Metric 1: Test Substance Identity	High	x 2 2
Metric 2: Test Substance Source	Medium	X 1	2 Two source companies indicated, but not specific for
methylene chloride.
Metric 3: Test Substance Purity	Low	X 1	3 Source company indicated, suggests at least a tech-
nical grade; but no purity given.
Domain 2: Test Design
Metric 4: Negative Controls	High	x 2 2
Metric 5: Negative Control Response	Low	X 1	3 Results reported relative to controls in the devel-
opmental test, but negative control results not pro-
vided.
Metric 6: Randomized Allocation	Low	X 1	3 Randomized allocation not indicated.
Domain 3: Exposure Characterization
Metric 7: Experimental System/Test Media Prepara-
tion
Metric 8: Consistency of Exposure Administration
Metric 9: Measurement of Test Substance Concentra-
tion
Metric 10: Exposure Duration and Frequency
Metric 11: Number of Exposure Groups/Spacing of Ex-
posure Levels
Metric 12: Testing at or Below Solubility Limit
Low
High
N/A
High
High
High
x 2
x 1
x 2
x 1
x 1
6
1
N/A
DCM's volatility was not addressed.
Testing conducted in a medium that would interfere
with analytical instruments. Effects were observed
at low concentrations suggesting DCM was main-
tained in the medium.
Domain 4: Test Organism
Metric 13: Test Organism Characteristics
Unacceptable x 2
Metric 14: Acclimitization and Pretreatment Conditions High
x 1
The nematode is not a well-known laboratory
species, and is also not a freshwater species, lives
in beer mats; therefore, not relevant for aquatic tox-
icity.
Continued on next page . ..
11

-------
. continued from previous page
Study Citation: Samoiloff, M.R., Schulz, S., Jordan, Y., Denich, K., Arnott, E.. 1980. A rapid simple long-term toxicity assay for aquatic contaminants
using the nematode Panagrellus redivivus. Can. J. Fish. Aquat. Sci. 37:1167-1174
Data Type:	Acute (0-96 hour); Aquatic; Invertebrates
Hero ID:	29147
Domain
Metric
Rating1
MWF* Score
Comments' t
Metric 15: Number of Organisms and Replicates per High	x 1	1
Group
Metric 16: Adequacy of Test Conditions	Medium	x 1	2
No details on biomass loading, temperature, etc were
provided.
Domain 5: Outcome Assessment
Metric 17: Outcome Assessment Methodology
Metric 18: Consistency of Outcome Assessment
High
High
x 2
x 1
Domain 6: Confounding / Variable Control
Metric 19:
Confounding Variables in Test Design and
Low
X
2
6
Study did not provide information on individual en-

Procedures




vironmental conditions for each chemical tested to





allow a comparison.
Metric 20:
Outcomes Unrelated to Exposure
Low
X
1
3
Control health outcomes not indicated.
Domain 7: Data Presentation and Analysis





Metric 21:
Statistical Methods
High
X
1
1

Metric 22:
Reporting of Data
Medium
X
2
4
Control results not provided.
Metric 23:
Explanation of Unexpected Outcomes
High
X
1
1

Overall Quality Determination"'"
Unacceptable
4.0 Metric mean score**: 1.9.
Extracted
No


** Consistent with our Application of Systematic Review in TSCARisk Evaluations document, if a metric for a data source receives a score of Unacceptable (score = 4),
EPA will determine the study to be unacceptable. In this case, one of the metrics were rated as unacceptable. As such, the study is considered unacceptable and the score
is presented solely to increase transparency.
*	MWF = Metric Weighting Factor
t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value.
*	The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High.
Overall rating =
(Metric Score; x MWF;) / ^ MWFj
if any metric is Unacceptable
(round to the nearest tenth) otherwise
where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed
out and an arrow points to the new rating.
^ Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments.
12

-------
Study Citation: Geiger, D.
L.,Poirier, S. H.,Brooke, L. T.,Call, D. J. eds.
1986. Acute toxicities of organic chemicals to fathead minnows (Pimephales
promelas):
volume III.




Data Type: Acute (0-96 hour); Aquatic; Fish




Hero ID: 32170





Domain
Metric
Rating^
MWF*
Score
Comments^ f
Domain 1: Test Substance





Metric 1:
Test Substance Identity
High
x 2
2
Test substance was identified as dichloromethane.
Metric 2:
Test Substance Source
High
x 1
1
Test substance source was Aldrich Chemical Com-





pany.
Metric 3:
Test Substance Purity
High
x 1
1
Purity was 99+ percent for dichloromethane.
Domain 2: Test Design





Metric 4:
Negative Controls
High
x 2
2
Lake Superior water or municipal city water used for





controls.
Metric 5:
Negative Control Response
Low
x 1
3
Control behavioral effects and mortality data were





reported.
Metric 6:
Randomized Allocation
Medium
x 1
2
Random distribution of test fish indicated but ran-





domization method not described.
Domain 3: Exposure Characterization




Metric 7:
Experimental System/Test Media Prepara-
Low
x 2
6
Authors noted high volatility of methylene chloride

tion



but did not cap test vessels.
Metric 8: Consistency of Exposure Administration
Metric 9: Measurement of Test Substance Concentra- High
tion
Metric 10: Exposure Duration and Frequency	High
Metric 11: Number of Exposure Groups/Spacing of Ex- High
posure Levels
Metric 12: Testing at or Below Solubility Limit
Medium X 1	2 Flow-through exposures were made with various sys-
tems described by the authors, but exposure system
for dichloromethane was not specified.
X 1	1	Gas-liquid chromatography was used to measure
dichloromethane daily in test vessels.
X 2	2 The tests were flow-through exposures for 96-hours,
which is adequate for detection of effects and mor-
tality in fish.
X 1	1	Five exposure concentrations and the control, rang-
ing from 0 to 1120 mg/L nominally, were adequately
spaced to develop a concentration-response curve.
X 1	1	Testing was below the water solubility limit for
dichloromethane.
High
Domain 4: Test Organism
Continued on next page . . .
13

-------
. . . continued from previous page
Study Citation:
Geiger, D. L.,Poirier, S. H.,Brooke, L. T.,Call, D. J. eds. 1986. Acute toxicities of organic chemicals to fathead minnows (Pimephales

promelas): volume III.
Data Type:
Acute (0-96 hour); Aquatic; Fish
Hero ID:
32170
Domain
Metric Rating"!" MWF* Score Comments"*1"!"
The test organism is a well-known species,
Pimephale promelas (fathed minnow), cultured in
U.S. EPA lab or at U. of Wisconsin.
Acclimatization and pretreatment conditions were
similar to test conditions, minus feeding 24 hours
prior to and during testing.
For the single cell, electronic, or mini exposure sys-
tems, 5, 10, or 20 fish per treatment and controls
were used, respectively.
Loading rate was less than 0.5 g/L/day per test
vessel and environmental conditions were monitored
and similar to pre-test conditions.
Metric 13: Test Organism Characteristics	High	x 2
Metric 14: Acclimitization and Pretreatment Conditions	High	x 1
Metric 15: Number of Organisms and Replicates per	High	x 1
Group
Metric 16: Adequacy of Test Conditions	High	x 1
Domain 5: Outcome Assessment
Metric 17: Outcome Assessment Methodology	High	X 2	2 For dichloromethane, dead fish were noted and re-
moved daily, and fish behavior was noted using de-
tailed criteria. The authors used the corrected aver-
age of tank test concentrations in analysis.
Metric 18: Consistency of Outcome Assessment	High x 1	1
Domain 6: Confounding / Variable Control
Metric 19: Confounding Variables in Test Design and High	X 2	2 Environmental conditions were consistent across test
Procedures	groups for dichloromethane. There were no behav-
ioral effects or mortalities reported in the controls.
Metric 20: Outcomes Unrelated to Exposure	High	X 1	1 There were no behavioral effects or morality re-
ported in controls.
Domain 7: Data Presentation and Analysis





Metric 21:
Statistical Methods
High
X
1
1
The authors used Trimmed Spearman-Karber






method and corrected average of tank concentra-






tions to assess EC50s and LC50s with 95 percent






confidence intervals.
Metric 22:
Reporting of Data
High
X
2
2

Metric 23:
Explanation of Unexpected Outcomes
High
X
1
1

Overall Quality Determination"!"

High


1.3

Continued on next page . . .
14

-------
. . . continued from previous page
Study Citation: Geiger, D. L.,Poirier, S. H.,Brooke, L. T.,Call, D. J. eds. 1986. Acute toxicities of organic chemicals to fathead minnows (Pimephales
promelas): volume III.
Data Type:	Acute (0-96 hour); Aquatic; Fish
Hero ID:	32170
Domain
Metric
Rating"!" MWF* Score
Comments'^
Extracted
Yes
* MWF = Metric Weighting Factor
"!" High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value.
^ The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High.
Overall rating =
(Metric Score; X MWF;) / ]T\ MWF,
if any metric is Unacceptable
(round to the nearest tenth) otherwise
where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is
crossed out and an arrow points to the new rating,
tt Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments.
15

-------
Study Citation: Alexander, H. C.,McCarty, W. M.,Bartlett, E. A.. 1978. Toxicity of perchloroethylene, trichloroethylene, 1,1,1-trichloroethane, and
methylene chloride to fathead minnows. Bulletin of Environmental Contamination and Toxicology 20:344-352
Data Type:	Acute (0-96 hour); Aquatic; other Fish; Static Test
Hero ID:	58126
Domain
Metric
Rating"!" MWF* Score
Comments^ f
Domain 1: Test Substance






Metric 1:
Test Substance Identity
High
X
2
2
Test substance identified as methylene chlorid
Metric 2:
Test Substance Source
Medium
X
1
2
Substance source not identified, however, the au-






thors work for chemical company that produces






methylene chloride.
Metric 3:
Test Substance Purity
Low
X
1
3
Test substance purity was not indicated.
Domain 2: Test Design






Metric 4:
Negative Controls
High
X
2
2
A negative control with dechlorinated lake water was
Metric 5:
Negative Control Response
Low
X
1
3
A negative control response was not reported.
Metric 6:
Randomized Allocation
Low
X
1
3
Randomization was not indicated.
Domain 3: Exposure Characterization





Metric 7:
Experimental System/Test Media Prepara-
Low
X
2
6
Test chambers were covered with plastic wrap, but

tion




no indication that headspace minimized to reduce





volatilization of the test substance.
Metric 8:
Consistency of Exposure Administration
High
X
1
1

Metric 9:
Measurement of Test Substance Concentra-
Low
X
1
3
Nominal concentrations were used in the static test,

tion




and some loss of methylene chloride is expected from





vaporization into the headspace of the plastic cov-






ered vessels.
Metric 10:
Exposure Duration and Frequency
High
X
2
2
This was a 96-hour test adequate for determining






the median lethal concentration (LC50) in fish.
Metric 11:
Number of Exposure Groups/Spacing of Ex-
Low
X
1
3
Test concentrations and spacing of exposure concen-

posure Levels




trations was not provided.
Metric 12:
Testing at or Below Solubility Limit
Medium
X
1
2
Test concentrations were not provided, but test re-






sults were below the water solubility limit for methy-






lene chloride.
Domain 4: Test Organism






Metric 13:
Test Organism Characteristics
Low
X
2
6
Test organisms (fathead minnow) were obtained






from a bait company, not cultured for laboratory






testing.
Continued on next page
16

-------
. . . continued from previous page
Study Citation: Alexander, H. C.,McCarty, W. M.,Bartlett, E. A.. 1978. Toxicity of perchloroethylene, trichloroethylene, 1,1,1-trichloroethane, and
methylene chloride to fathead minnows. Bulletin of Environmental Contamination and Toxicology 20:344-352
Data Type:	Acute (0-96 hour); Aquatic; other Fish; Static Test
Hero ID:	58126
Domain
Metric
Rating^
MWF*
Score
Comments^
Metric 14:
Acclimitization and Pretreatment Conditions
Low
X
1
3
Test organisms from a bait company which leaves






uncertainty regarding disease, age, and origin of the






fish; however, control and test outcomes do not ap-






pear to be impacted.
Metric 15:
Number of Organisms and Replicates per
Low
X
1
3
Number of organisms and replicates per exposure

Group




concentration were not indicated.
Metric 16:
Adequacy of Test Conditions
Low
X
1
3
Details on test conditions for the static test were not






provided.
Domain 5: Outcome Assessment





Metric 17:
Outcome Assessment Methodology
High
X
2
2
Dead or affected fish were counted and dead fish were






removed from test vessels on a daily basis.
Metric 18:
Consistency of Outcome Assessment
High
X
1
1

Domain 6: Confounding / Variable Control





Metric 19:
Confounding Variables in Test Design and
Low
X
2
6
No details regarding the environmental conditions

Procedures




for each exposure concentration or the controls were





provided.
Metric 20:
Outcomes Unrelated to Exposure
Low
X
1
3
No information on outcomes unrelated to exposures






was provided.
Domain 7: Data Presentation and Analysis





Metric 21:
Statistical Methods
High
X
1
1
Probit analyis was used to obtain LC10, LC50 and






LC90 with 95 percent confidence limits.
Metric 22:
Reporting of Data
Medium
X
2
4
Results for Flow-through tests were compared to






static, but control responses not provided.
Metric 23:
Explanation of Unexpected Outcomes
Low
X
1
3
No details on unexpected outcomes were provided.
Overall Quality Determination"'"
Medium
2.2
Extracted
Yes


Continued on next page . . .
17

-------
. continued from previous page
Study Citation: Alexander, H. C.,McCarty, W. M.,Bartlett, E. A.. 1978. Toxicity of perchloroethylene, trichloroethylene, 1,1,1-trichloroethane, and
methylene chloride to fathead minnows. Bulletin of Environmental Contamination and Toxicology 20:344-352
Data Type:	Acute (0-96 hour); Aquatic; other Fish; Static Test
Hero ID:	58126
Domain
Metric
Rating"'" MWF* Score
Comments^

*	MWF = Metric Weighting Factor
t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value.
*	The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High.
Overall rating =
^ (Metric Score; x MWF;) / ^ MWFj
if any metric is Unacceptable
(round to the nearest tenth) otherwise
where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is
crossed out and an arrow points to the new rating.
^ Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments.
18

-------
Study Citation: Alexander, H. C.,McCarty, W. M.,Bartlett, E. A.. 1978. Toxicity of perchloroethylene, trichloroethylene, 1,1,1-trichloroethane, and
methylene chloride to fathead minnows. Bulletin of Environmental Contamination and Toxicology 20:344-352
Data Type:	Acute (0-96 hour); Aquatic; other Fish; Flow-Through Test
Hero ID:	58126
Domain	Metric	Rating^ MWF* Score	Comments^
Domain 1: Test Substance
Metric 1:
Metric 2:
Test Substance Identity
Test Substance Source
Metric 3: Test Substance Purity
High	X 2	2 Test substance identified as methylene chloride.
Medium x 1	2 Substance source not identified, however authors
work for chemical company that produces methylene
chloride.
X 1	3	Test substance purity was not indicated.
Low
Domain 2: Test Design
Metric 4:
Negative Controls
High
X
2
2
Controls were used (lake water).
Metric 5:
Negative Control Response
Low
X
1
3
A negative control response was not reported
Metric 6:
Randomized Allocation
Low
X
1
3
Randomization not indicated.
Domain 3: Exposure Characterization
Metric 7: Experimental System/Test Media Prepara- Low
tion
Metric 8: Consistency of Exposure Administration	High
Metric 9: Measurement of Test Substance Concentra- Medium
tion
Metric 10: Exposure Duration and Frequency
High
x 2
x 1
x 1
x 2
x 1
Metric 11: Number of Exposure Groups/Spacing of Ex- Low
posure Levels
Metric 12: Testing at or Below Solubility Limit	Medium x 1
Covered with plastic wrap, but no indication that
headspace minimized.
Gas chromatography used to analyze test concentra-
tions daily for the flow-through test, but few details
about methodology provided.
This 96-hour test was adequate duration for deter-
mining the median lethal concentration in fish.
Test concentrations and spacing of exposure concen-
trations was not provided.
Test concentrations were not provided, but test re-
sults were below the water solubility limit for methy-
lene chloride.
Domain 4: Test Organism
Metric 13: Test Organism Characteristics	Low	X 2	6 Test organisms were obtained from a bait company,
not cultured for laboratory testing.
Continued on next page . . .
19

-------
. . . continued from previous page
Study Citation: Alexander, H. C.,McCarty, W. M.,Bartlett, E. A.. 1978. Toxicity of perchloroethylene, trichloroethylene, 1,1,1-trichloroethane, and
methylene chloride to fathead minnows. Bulletin of Environmental Contamination and Toxicology 20:344-352
Data Type:	Acute (0-96 hour); Aquatic; other Fish; Flow-Through Test
Hero ID:	58126
Domain
Metric
Rating^
MWF*
Score
Comments^
Metric 14:
Acclimitization and Pretreatment Conditions
Low
X
1
3
Test organisms were obtained from a bait company,






which leaves uncertainty regarding origin (purity






and consistency of stock), age, and disease of the






fish; however, control results appeared normal so






this does not appear to impact test results.
Metric 15:
Number of Organisms and Replicates per
Low
X
1
3
The number of organisms and replicates per test con-

Group




centration was not reported.
Metric 16:
Adequacy of Test Conditions
Low
X
1
3
Although water quality parameters (pH, D.O, tem-






purature, etc.) were provided prior to test initiation,






details on test conditions (biomass loading, aquaria






size, water quality during and after testing) were not






provided.
Domain 5: Outcome Assessment





Metric 17:
Outcome Assessment Methodology
High
X
2
2
Dead or affected fish were counted and dead fish were






removed from test vessels each day in order to assess






the median lethal concentration.
Metric 18:
Consistency of Outcome Assessment
High
X
1
1

Domain 6: Confounding / Variable Control





Metric 19:
Confounding Variables in Test Design and
Low
X
2
6
No details regarding the environmental conditions

Procedures




during testing were provided.
Metric 20:
Outcomes Unrelated to Exposure
Low
X
1
3
No details on outcomes unrelated to exposures were






provided.
Domain 7: Data Presentation and Analysis





Metric 21:
Statistical Methods
High
X
1
1
Probit analysis was used to obtain LC10, LC50, and






LC90 with 95 percent confidence limits.
Metric 22:
Reporting of Data
High
X
2
2

Metric 23:
Explanation of Unexpected Outcomes
Low
X
1
3
No details on unexpected outcomes provided.
Overall Quality Determination"'"
Medium
2.1
Extracted
Yes


Continued on next page . . .
20

-------
. continued from previous page
Study Citation: Alexander, H. C.,McCarty, W. M.,Bartlett, E. A.. 1978. Toxicity of perchloroethylene, trichloroethylene, 1,1,1-trichloroethane, and
methylene chloride to fathead minnows. Bulletin of Environmental Contamination and Toxicology 20:344-352
Data Type:	Acute (0-96 hour); Aquatic; other Fish; Flow-Through Test
Hero ID:	58126
Domain
Metric
Rating"'" MWF* Score
Comments^

*	MWF = Metric Weighting Factor
t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value.
*	The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High.
Overall rating =
^ (Metric Score; x MWF;) / ^ MWFj
if any metric is Unacceptable
(round to the nearest tenth) otherwise
where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is
crossed out and an arrow points to the new rating.
^ Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments.
21

-------
Study Citation: Kuhn, R.,Pattard, M.,Pernak, K. D.,Winter, A.. 1989. Results of the harmful effects of selected water pollutants (anilines, phenols,
aliphatic compounds) to Daphnia magna. Water Research 23:495-499
Data Type:	Acute (0-96 hour); Aquatic; Invertebrates
Hero ID:	85242
Domain	Metric	Rating^ MWF* Score	Comments^
Domain 1: Test Substance
Metric 1:
Test Substance Identity
High
X
2
2

Metric 2:
Test Substance Source
Low
X
1
3
Multi-chemical test, no source indicated.
Metric 3:
Test Substance Purity
Low
X
1
3
No purity or grade information provided
Domain 2: Test Design
Metric 4:
Metric 5:
Metric 6:
Negative Controls
Negative Control Response
Randomized Allocation
High
Medium
Low
x 2
x 1
x 1
Control response not reported, but only used con-
trols with <10 percent inhibition.
Randomization not indicated.
Domain 3: Exposure Characterization
Metric 7: Experimental System/Test Media Prepara-
tion
Metric 8: Consistency of Exposure Administration
Metric 9: Measurement of Test Substance Concentra-
tion
Metric 10: Exposure Duration and Frequency
Metric 11: Number of Exposure Groups/Spacing of Ex-
posure Levels
Metric 12: Testing at or Below Solubility Limit
Medium x 2
High
Low
High
Medium
Low
x 1
x 1
x 2
x 1
x 1
Closed vessel with ground-glass stoppers, but did
not reduce headspace to further reduce volatiliza-
tion.
Nominal concentrations for a volatile substance and
no analytical monitoring. Headspace in stoppered
vessels will still have volatilization.
Specific concentrations not provided for methylene
chloride, but range tested to provide 3-4 EC values
from ECO to EC100 and EC50.
Specific concentrations not provided for methylene
chloride to determine this.
Domain 4: Test Organism
Metric 13:
Test Organism Characteristics
High
X
2
2
Metric 14:
Acclimitization and Pretreatment Conditions
High
X
1
1
Metric 15:
Number of Organisms and Replicates per
High
X
1
1

Group




Metric 16:
Adequacy of Test Conditions
High
X
1
1
Continued on next page . . .
22

-------
. .. continued from previous page
Study Citation: Kuhn, R.,Pattard, M.,Pernak, K. D.,Winter, A.. 1989. Results of the harmful effects of selected water pollutants (anilines, phenols,
aliphatic compounds) to Daphnia magna. Water Research 23:495-499
Data Type:	Acute (0-96 hour); Aquatic; Invertebrates
Hero ID:	85242
Domain Metric
Rating^
MWF*
Score
Comments^

Domain 5: Outcome Assessment





Metric 17: Outcome Assessment Methodology
High
X
2
2

Metric 18: Consistency of Outcome Assessment
High
X
1
1

Domain 6: Confounding / Variable Control





Metric 19: Confounding Variables in Test Design and
Medium
X
2
4
Study did not provide environmental information for
Procedures




comparison, but authors indicated that DO and pH




were evaluated at the end and factored into the re-





sults.
Metric 20: Outcomes Unrelated to Exposure
Low
X
1
3
Health outcomes for controls not reported, but au-





thors indicated that controsl with >10 percent inhi-





bition were not used.
Domain 7: Data Presentation and Analysis





Metric 21: Statistical Methods
High
X
1
1

Metric 22: Reporting of Data
Medium
X
2
4
Endpoint values and only use of controls with <10





percent inhibition were reported, but no other de-





tails provided.
Metric 23: Explanation of Unexpected Outcomes
High
X
1
1

Overall Quality Determination"'"
Modium --> Low
1.7
Study did not provide substance purity, source,
or test concentration, and headspace in the vessel
allowed volatilization of methylene chloride.
Extracted
Yes



*	MWF = Metric Weighting Factor
t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value.
*	The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High.
Overall rating =
(Metric Score; x MWF;) / ^ MWFj
if any metric is Unacceptable
(round to the nearest tenth) otherwise
where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is
crossed out and an arrow points to the new rating.
Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments.
23

-------
Study Citation: Black, J. A.,Birge, W. J.,McDonnell, W. E.,Westerman, A. G.,Ramey, B. A.,Bruser, D. M.. 1982. The aquatic toxicity of organic
compounds to embryo-larval stages of fish and amphibians. 133
Data Type:	Acute (0-96 hour); Aquatic; other Amphibians
Hero ID:	93660
Domain	Metric	Rating^ MWF* Score	Comments^
Domain 1: Test Substance
Metric 1:
Test Substance Identity
High
X
2
2
Test substance was identified as methylene chloride
Metric 2:
Test Substance Source
Low
X
1
3
Substance source was not indicated.
Metric 3:
Test Substance Purity
High
X
1
1
All test substances were reagent grade.
Domain 2: Test Design
Metric 4:
Negative Controls
High
X
2
2
Fish and amphibian control eggs were used.
Metric 5:
Negative Control Response
High
X
1
1
Control survival ranged from 84-99 percent.
Metric 6:
Randomized Allocation
Low
X
1
3
There was no mention of randomized allocation of
test organisms.
Domain 3: Exposure Characterization
Metric 7: Experimental System/Test Media Prepara- High
tion
Metric 8: Consistency of Exposure Administration	High
Metric 9: Measurement of Test Substance Concentra- High
tion
Metric 10: Exposure Duration and Frequency	High
Metric 11: Number of Exposure Groups/Spacing of Ex- High
posure Levels
Metric 12: Testing at or Below Solubility Limit	High
x 2
x 1
x 1
x 2
x 1
x 1
Flow-through testing with closed vessel devoid of air
space was used to minimize volatilization.
Gas-liquid chromatography was used to measure test
concentrations daily.
Amphibian embryo-larvae were exposed up to 4 days
post-hatch, sufficient to determine effects in embryos
and larvae.
There were 6 exposure concentrations with appro-
priate spacing used fore each amphibian tested.
All exposure concentrations were below the water
solubility of methylene chloride.
Domain 4: Test Organism
Metric 13: Test Organism Characteristics
High
x 2
Metric 14: Acclimitization and Pretreatment Conditions High x 1
Amphibians used were appropriate for this study,
with the exception of the African Clawed frog, which
is not endemic to the U.S.
Freshly fertilized eggs from amphibians were either
obtained from laboratory suppliers or from amphib-
ians cultured in the lab.
Continued on next page . . .
24

-------
. continued from previous page
Study Citation: Black, J. A.,Birge, W. J.,McDonnell, W. E.,Westerman, A. G.,Ramey, B. A.,Bruser, D. M.. 1982. The aquatic toxicity of organic
compounds to embryo-larval stages of fish and amphibians. 133
Data Type:	Acute (0-96 hour); Aquatic; other Amphibians
Hero ID:	93660
Domain
Metric
Rating"!" MWF* Score
Comments'^
Metric 15: Number of Organisms and Replicates per High x 1	1
Group
Metric 16: Adequacy of Test Conditions	High x 1	1
Single replicates of 50 to 125 eggs were used per test
concentration.
A loading rat of up to 125 eggs per test concen-
tration was used, which did not appear to impact
test results. Environmental conditions were within
acceptable ranges, and control mortality was accept-
able.
Domain 5: Outcome Assessment
Metric 17: Outcome Assessment Methodology
Metric 18: Consistency of Outcome Assessment
High	X 2	2 Test vessels observed daily to assess development
and remove dead test organisms.
Medium X 1	2 LC50, LCIO, LCls were assessed adjusted for control
mortality, but detailed control mortality data were
not provided.
Domain 6: Confounding / Variable Control
Metric 19: Confounding Variables in Test Design and High x 2
Procedures
Metric 20: Outcomes Unrelated to Exposure
Medium x 1
Environmental conditions appeared consistent
across test concentrations and control mortality
ranged from 1-16 percent.
Teratogenesis was reportedly infrequent in controls
( percent teratogenicity not reported) and control
mortality ranged from 1 to 16 percent, which is ac-
ceptable.
Domain 7: Data Presentation and Analysis
Metric 21: Statistical Methods
Metric 22: Reporting of Data
Metric 23: Explanation of Unexpected Outcomes
Medium x 1
Medium x 2
High x 1
Survivability was reported as percent of total organ-
isms at each exposure concentration after corrected
for control mortality, but detailed control data were
not reported. LC50s, LClOs, and LCls were calcu-
lated using log-probit analysis.
All dose-response and survivability data were re-
ported after correction for control mortality, but
control data was not reported.
Overall Quality Determination"'"
High
1.3
Continued on next page . . .
25

-------
. continued from previous page
Study Citation: Black, J. A.,Birge, W. J.,McDonnell, W. E.,Westerman, A. G.,Ramey, B. A.,Bruser, D. M.. 1982. The aquatic toxicity of organic
compounds to embryo-larval stages of fish and amphibians. 133
Data Type:	Acute (0-96 hour); Aquatic; other Amphibians
Hero ID:	93660
Domain
Metric
Rating"!" MWF* Score
Comments'^
Extracted
Yes
* MWF = Metric Weighting Factor
"!" High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value.
^ The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High.
Overall rating =
(Metric Score; X MWF;) / J] . MWF,
if any metric is Unacceptable
(round to the nearest tenth) otherwise
where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is
crossed out and an arrow points to the new rating.
Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments.
26

-------
Study Citation: Black, J. A.,Birge, W. J.,McDonnell, W. E.,Westerman, A. G.,Ramey, B. A.,Bruser, D. M.. 1982. The aquatic toxicity of organic
compounds to embryo-larval stages of fish and amphibians. 133
Data Type:	Acute (0-96 hour); Aquatic; Fish
Hero ID:	93660
Domain	Metric	Rating^ MWF* Score	Comments^
Domain 1: Test Substance
Metric 1:
Test Substance Identity
High
X
2
2
Test substance was identified as methylene chloride
Metric 2:
Test Substance Source
Low
X
1
3
Substance source was not indicated.
Metric 3:
Test Substance Purity
High
X
1
1
All test substances were reagent grade.
Domain 2: Test Design
Metric 4:
Negative Controls
High
X
2
2
Fish and amphibian control eggs were used.
Metric 5:
Negative Control Response
High
X
1
1
Control survival ranged from 84-99 percent.
Metric 6:
Randomized Allocation
Low
X
1
3
There was no mention of randomized allocation of
test organisms.
Domain 3: Exposure Characterization
Metric 7: Experimental System/Test Media Prepara- High
tion
Metric 8: Consistency of Exposure Administration	High
Metric 9: Measurement of Test Substance Concentra- High
tion
Metric 10: Exposure Duration and Frequency	High
Metric 11: Number of Exposure Groups/Spacing of Ex- High
posure Levels
Metric 12: Testing at or Below Solubility Limit	High
x 2
x 1
x 1
x 2
x 1
x 1
Flow-through testing with closed vessel devoid of air
space was used to minimize volatilization.
Gas-liquid chromatography was used to measure test
concentrations daily.
Fish embryo-larvae were exposed up to 4 days post-
hatch , sufficient to determine effects in embryos and
larvae.
There were 6 exposure concentrations with appro-
priate spacing used for each fish tested.
All exposure concentrations were below the water
solubility of methylene chloride.
Domain 4: Test Organism
Metric 13:
Test Organism Characteristics
High
x 2
Rainbow trout and fathead minnow are well known
species. The trout were obtained from a hatchery
and freshly fertilized fathead minnow eggs were ob-
tained from the EPA Newtown Fish Toxicology Lab-
oratory.
Continued on next page
27

-------
. continued from previous page
Study Citation: Black, J. A.,Birge, W. J.,McDonnell, W. E.,Westerman, A. G.,Ramey, B. A.,Bruser, D. M.. 1982. The aquatic toxicity of organic
compounds to embryo-larval stages of fish and amphibians. 133
Data Type:	Acute (0-96 hour); Aquatic; Fish
Hero ID:	93660
Domain
Metric
Rating"!" MWF* Score
Comments'^
Metric 14: Acclimitization and Pretreatment Conditions	High	x	1	1
Metric 15: Number of Organisms and Replicates per	High	x	1	1
Group
Metric 16: Adequacy of Test Conditions	High	x	1	1
Freshly fertilized eggs from fish were either obtained
from laboratory suppliers or from fish cultured in
the lab.
Single replicates of 50 to 125 eggs were used per test
concentration.
A loading rat of up to 125 eggs per test concen-
tration was used, which did not appear to impact
test results. Environmental conditions were within
acceptable ranges, and control mortality was accept-
able.
Domain 5: Outcome Assessment
Metric 17: Outcome Assessment Methodology
Metric 18: Consistency of Outcome Assessment
High	X 2	2 Test vessels observed daily to assess development
and remove dead test organisms.
Medium X 1	2 LC50, LCIO, LCls were assessed adjusted for control
mortality, but detailed control mortality data were
not provided.
Domain 6: Confounding / Variable Control
Metric 19: Confounding Variables in Test Design and High x 2
Procedures
Metric 20: Outcomes Unrelated to Exposure
Medium x 1
Environmental conditions appeared consistent
across test concentrations and control mortality
ranged from 1-16 percent.
Teratogenesis was reportedly infrequent in controls
( percent teratogenicity not reported) and control
mortality ranged from 1 to 16 percent, which is ac-
ceptable.
Domain 7: Data Presentation and Analysis
Metric 21: Statistical Methods
Metric 22: Reporting of Data
Metric 23: Explanation of Unexpected Outcomes
Medium x 1
Medium x 2
High x 1
Survivability was reported as percent of total organ-
isms at each exposure concentration after corrected
for control mortality, but detailed control data were
not reported. LC50s, LClOs, and LCls were calcu-
lated using log-probit analysis.
All dose-response and survivability data were re-
ported after correction for control mortality, but
control data was not reported.
Continued on next page . . .
28

-------
. . . continued from previous page
Study Citation: Black, J. A.,Birge, W. J.,McDonnell, W. E.,Westerman, A. G.,Ramey, B. A.,Bruser, D. M.. 1982. The aquatic toxicity of organic
compounds to embryo-larval stages of fish and amphibians. 133
Data Type:	Acute (0-96 hour); Aquatic; Fish
Hero ID:	93660
Domain	Metric	Rating^ MWF* Score	Comments^
Overall Quality Determination"'"	High	1.3
Extracted	Yes
* MWF = Metric Weighting Factor
t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value.
t The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High.
if any metric is Unacceptable
Overall rating =
(Metric Score; X MWF;) / ]T\ MWF,
(round to the nearest tenth) otherwise
where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is
crossed out and an arrow points to the new rating.
It Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments.
29

-------
Study Citation: Sanchez-Fortun, S.,Sanz, F.,Santa-Maria, A.,Ros, J. M.,De Vicente, M. L.,Encinas, M. T.,Vinagre, E.,Barahona, M. V.. 1997. Acute
sensitivity of three age classes of Artemia salina larvae to seven chlorinated solvents. Bulletin of Environmental Contamination and
Toxicology 59:445-451
Data Type:	Acute (0-96 hour); Aquatic; Invertebrates
Hero ID:	200570
Domain	Metric	Rating"!" MWF* Score	Comments"*1"!"
Domain 1: Test Substance
Metric 1:
Test Substance Identity
High
X
2
2
Test substance identified as methylene chloride.
Metric 2:
Test Substance Source
High
X
1
1
Test substance obtained from Sigma Chemical Company.
Metric 3:
Test Substance Purity
High
X
1
1
The test substance wa identifieda as analytical grade.
Test Design


X



Metric 4:
Negative Controls
High
2
2
A <1% ethanol control and synthetic seawater control were used.
Metric 5:
Negative Control Response
Low
X
1
3
The control responses were not reported.
Metric 6:
Randomized Allocation
Low
X
1
3
Randomized allocation was not indicated.
Domain 3: Exposure Characterization
Metric 7: Experimental System/Test Media Prepara-
tion
Metric 8: Consistency of Exposure Administration
Metric 9: Measurement of Test Substance Concentra-
tion
Unacceptable x 2
High x i
Unacceptable x 1
Nominal concentrations used without steps to reduce
volatilization of methylene chloride.
Nominal concentrations were reported and methylene
chloride is volatile.
Metric 10: Exposure Duration and Frequency	Medium x 2 4
Metric 11: Number of Exposure Groups/Spacing of Ex- Low	x 1	3
posure Levels
A 24-hour exposure period is not adequate to capture effects in
D. magna based on OCSPP TG 850.1010.
Study does not provide exposure concentrations, but
six replicates were used and LC50/EC50s indicate
exposure concentrations were sufficiently spaced.
Continued on next page . . .
30

-------
. . . continued from previous page
Study Citation: Sanchez-Fortun, S.,Sanz, F.,Santa-Maria, A.,Ros, J. M.,De Vicente, M. L.,Encinas, M. T.,Vinagre, E.,Barahona, M. V.. 1997. Acute
sensitivity of three age classes of Artemia salina larvae to seven chlorinated solvents. Bulletin of Environmental Contamination and
Toxicology 59:445-451
Data Type:	Acute (0-96 hour); Aquatic; Invertebrates
Hero ID:	200570
Domain
Metric
Rating^
MWF*
Score
Comments'r
Metric 12:
Testing at or Below Solubility Limit
High
X
1
1

Domain 4: Test Organism






Metric 13:
Test Organism Characteristics
High
X
2
2
The test organism was Artemia salina, a saltwater invertebrate .
Metric 14:
Acclimitization and Pretreatment Conditions
High
X
1
1

Metric 15:
Number of Organisms and Replicates per
High
X
1
1
10 animals and four replicates per methylene chlo-

Group




ride concentration tested.
Metric 16:
Adequacy of Test Conditions
High
X
1
1

Domain 5: Outcome Assessment





Metric 17:
Outcome Assessment Methodology
High
X
2
2
The median lethal concentration was assessed.
Metric 18:
Consistency of Outcome Assessment
High
X
1
1

Domain 6: Confounding / Variable Control





Metric 19:
Confounding Variables in Test Design and
High
X
2
2


Procedures





Metric 20:
Outcomes Unrelated to Exposure
Medium
X
1
2
Health outcomes unrelated to exposure (i.e. con-





trols) not reported, and minimal details were provided.
Domain 7: Data Presentation and Analysis





Metric 21:
Statistical Methods
High
X
1
1
The LC50 values, with 95 percent confidence limits,






were calculated using to Litchfield and Wilcoxon






method. Results were subjected to a two-way analysis of






variance with replication within the sub-groups, and






contrasted with the Newman-Keuls Test.
Metric 22:
Reporting of Data
Medium
X
2
4
Control results not provided, but unlikely to impact





results.
Metric 23:
Explanation of Unexpected Outcomes
High
X
1
1

Continued on next page . . .
31

-------
. . . continued from previous page
Study Citation:
Data Type:
Hero ID:
Sanchez-Fortun, S.,Sanz, F.,Santa-Maria, A.,Ros, J. M.,De Vicente, M. L.,Encinas, M. T.,Vinagre, E.,Barahona, M. V.. 1997. Acute
sensitivity of three age classes of Artemia salina larvae to seven chlorinated solvents. Bulletin of Environmental Contamination and
Toxicology 59:445-451
Acute (0-96 hour); Aquatic; Invertebrates
200570
Domain
Metric
Rating"!" MWF* Score Comments"^
Overall Quality Determination"'"

Unacceptable 4.0 Metric mean scoreEE: 1.7
Extracted

No

* MWF = Metric Weighting Factor
t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value.
The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High.
Overall rating =
(Metric Score; X MWF;) / J] . MWF,
if any metric is Unacceptable
(round to the nearest tenth) otherwise
where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is
crossed out and an arrow points to the new rating.
Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments.
32

-------
Study Citation:	Brack, W.,Rottler, H.. 1994. Toxicity testing of highly volatile chemicals with green algae: A new assay. 1:223-228
Data Type:	Acute (0-96 hour); Aquatic; Plants
Hero ID:	661061
Domain	Metric	Rating^ MWF* Score	Comments"^
Domain 1: Test Substance
Metric 1:
Test Substance Identity
High
X
2
2
Test substance was identified by name.
Metric 2:
Test Substance Source
High
X
1
1
Promochem was the source company for methylene
choride.
Metric 3:
Test Substance Purity
High
X
1
1
Purity was reported as nanograde.
Domain 2: Test Design
Metric 4:
Negative Controls
High
X
2
2
Indicated negative controls were used.
Metric 5:
Negative Control Response
High
X
1
1
Exponential growth was reported in control algae
Metric 6:
Randomized Allocation
Low
X
1
3
Randomization was not reported.
Domain 3: Exposure Characterization
Metric 7: Experimental System/Test Media Prepara- High x 2
tion
Metric 8: Consistency of Exposure Administration	High x 1
Metric 9: Measurement of Test Substance Concentra- High x 1
tion
Metric 10: Exposure Duration and Frequency	High x 2
Metric 11: Number of Exposure Groups/Spacing of Ex- High x 1
posure Levels
Metric 12: Testing at or Below Solubility Limit	High x 1
Headspace was removed in sealed test vials to mini-
mize volatilization.
Measurments by Gas chromatography/ECD after
liquid-liquid microextraction were taken at test ini-
tiation and end.
Algae were treated for 72 hours under static condi-
tions.
Seven treatment groups plus controls (positive and
negative) were tested. Test concentrations were not
reported, but are shown in the figures, which show
adequate spacing of the test concentrations to de-
termine concentration response curves for methylene
chloride.
Test concentrations were below the water solubility
limit for methylene chloride.
Domain 4: Test Organism
Metric 13: Test Organism Characteristics
Medium x 2
Metric 14: Acclimitization and Pretreatment Conditions High x 1
Chlamydomonas reinhardtii is a mobile algal species
with two flagella, not commonly used for toxicity
testing.
Continued on next page . . .
33

-------
. continued from previous page
Study Citation: Brack, W.,Rottler, H.. 1994. Toxicity testing of highly volatile chemicals with green algae: A new assay. 1:223-228
Data Type:	Acute (0-96 hour); Aquatic; Plants
Hero ID:	661061
Domain
Metric
Ratingt MWF* Score
Commentstt
Metric 15: Number of Organisms and Replicates per High x 1
Group
Metric 16: Adequacy of Test Conditions
High
x 1
Two replicates per test concentration and 3 negative
controls were reported with an initial inoculum of 5
x 103 cells/mL each. OECD test guideline recom-
mends 3 replicates unless a NOEC is not required,
which was the case here.
Culture conditions appear adequate and nega-
tive control results showed uninhibited exponential
growth.
Domain 5: Outcome Assessment





Metric 17:
Outcome Assessment Methodology
High
X
2
2
Biomass assessed using fluorometric measurement of






total chlorophyll for controls and treatment groups






to determined EClOs and EC50s.
Metric 18:
Consistency of Outcome Assessment
High
X
1
1
No inconsistencies were reported, and both positive






and negative controls performed as expected.
Domain 6: Confounding / Variable Control





Metric 19:
Confounding Variables in Test Design and
High
X
2
2
Both positive and negative controls performed as ex-

Procedures




pected, differences in environmental conditions were





not reported.
Metric 20:
Outcomes Unrelated to Exposure
High
X
1
1
Positive and negative controls performed as ex-






pected and no outcomes unrelated to exposures were






reported.
Domain 7: Data Presentation and Analysis





Metric 21:
Statistical Methods
High
X
1
1
Probit analysis was used to assess significant differ-






ences in biomass.
Metric 22:
Reporting of Data
Medium
X
2
4
Figure 3 shows the results of the tests at each cone






for each chemical but it's difficult to determine the






exact concentrations from the figure, so some minor






uncertainties remain.
Metric 23:
Explanation of Unexpected Outcomes
Medium
X
1
2
SDs were provided, but it was unclear whether or






not there were any unexpected outcomes.
Overall Quality Determination"'"
High


1.2

Extracted

Yes





Continued on next page . . .
34

-------
. continued from previous page
Study Citation: Brack, W.,Rottler, H.. 1994. Toxicity testing of highly volatile chemicals with green algae: A new assay. 1:223-228
Data Type:	Acute (0-96 hour); Aquatic; Plants
Hero ID:	661061
Domain	Metric	Rating^ MWF* Score	Comments'^
*	MWF = Metric Weighting Factor
t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value.
*	The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High.
if any metric is Unacceptable
Overall rating =
J]. (Metric Score; x MWF;) / J] . MWFj
(round to the nearest tenth) otherwise
where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is
crossed out and an arrow points to the new rating,
ft Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments.
35

-------
Study Citation:	Marquis, 0.,Millery, A.,Guittonneau, S.,Miaud, C.. 2006. Solvent toxicity to amphibian embryos and larvae. Chemosphere 63:889-892
Data Type:	Acute (0-96 hour); Aquatic; other Amphibians
Hero ID:	1042080
Domain	Metric	Rating^ MWF* Score	Comments"^
Domain 1: Test Substance
Metric 1:
Metric 2:
Metric 3:
Test Substance Identity
Test Substance Source
Test Substance Purity
High
Low
Low
x 2
x 1
x 1
Source not identified
Purity not identified
Domain 2: Test Design
Metric 4:
Metric 5:
Metric 6:
Negative Controls
Negative Control Response
Randomized Allocation
High
High
High
x 2
x 1
x 1
Domain 3: Exposure Characterization
Metric 7: Experimental System/Test Media Prepara-
tion
Metric 8: Consistency of Exposure Administration
Metric 9: Measurement of Test Substance Concentra-
tion
Metric 10: Exposure Duration and Frequency
Metric 11: Number of Exposure Groups/Spacing of Ex-
posure Levels
Metric 12: Testing at or Below Solubility Limit
Low
High
Low
High
x 2
Low	x 1
Unacceptable x 1
x 2
x 1
x 1
Volatility of the test material was not taken into ac-
count. The test concentrations were not measured
and endpoints reported in terms of nominal concen-
tration.
Exposure concentrations were not measured
Not all exposure groups were specified
Domain 4: Test Organism
Metric 13: Test Organism Characteristics
Medium
Metric 14: Acclimitization and Pretreatment Conditions	Low
Metric 15: Number of Organisms and Replicates per	High
Group
Metric 16: Adequacy of Test Conditions	High
x 2
x 1
x 1
x 1
Embryos were collected in the wild from agricultural
landscape, but no additional details about potential
exposure to pesticides was discussed. Lack of mor-
tality in controls indicates this wasn't an issue.
Acclimatization procedure was not discussed.
20
Continued on next page . . .
36

-------
. .. continued from previous page
Study Citation: Marquis, 0.,Millery, A.,Guittonneau, S.,Miaud, C.. 2006.
Data Type: Acute (0-96 hour); Aquatic; other Amphibians
Hero ID: 1042080
Solvent toxicity to amphibi
an embryos and larvae. Chemosphere 63:889-892
Domain Metric
Haling1
MWF*
Score
Commentstt
Domain 5: Outcome Assessment
Metric 17: Outcome Assessment Methodology
Metric 18: Consistency of Outcome Assessment
High
High
x 2
x 1
2
1

Domain 6: Confounding / Variable Control
Metric 19: Confounding Variables in Test Design and
Procedures
Metric 20: Outcomes Unrelated to Exposure
High
High
x 2
x 1
2
1

Domain 7: Data Presentation and Analysis
Metric 21: Statistical Methods
Metric 22: Reporting of Data
Metric 23: Explanation of Unexpected Outcomes
Low
Medium
High
x 1
x 2
x 1
3
4
1
Significance test methodology was not explained or
reported
Data summary was presented, whole dataset was not
presented
Overall Quality Determination"'"
Unacceptable

4.0
Metric mean score**: 1.7.
Extracted
No




** Consistent with our Application of Systematic Review in TSCARisk Evaluations document, if a metric for a data source receives a score of Unacceptable (score = 4),
EPA will determine the study to be unacceptable. In this case, one of the metrics were rated as unacceptable. As such, the study is considered unacceptable and the score
is presented solely to increase transparency.
*	MWF = Metric Weighting Factor
t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value.
*	The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High.
Overall rating =
^ (Metric Score; x MWF;) / ^ MWFj
if any metric is Unacceptable
(round to the nearest tenth) otherwise
where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed
out and an arrow points to the new rating,
ft Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments.
37

-------
Study Citation: Abernethy, S.,Bobra, A. M.,Shiu, W. Y.,Wells, P. G.,Mackay, D.. 1986. ACUTE LETHAL TOXICITY OF HYDROCARBONS
AND CHLORINATED HYDROCARBONS TO TWO PLANKTONIC CRUSTACEANS THE KEY ROLE OF ORGANISM-WATER
PARTITIONING. Aquatic Toxicology
Data Type:	Acute (0-96 hour); Aquatic; Invertebrates
Hero ID:	1486051
Domain	Metric	Rating"!" MWF* Score	Comments"*1"!"
Domain 1: Test Substance
Metric 1:
Test
Substance Identity
High
X
2
2
The test substance was identified as
dichloromethane.
Metric 2:
Test
Substance Source
Low
X
1
3
The study authors did not identify the test sub-
stance source.
Metric 3:
Test
Substance Purity
High
X
1
1
Test substance purity was reported as a minimum of
97 percent.
Domain 2: Test Design
Metric 4: Negative Controls
Metric 5: Negative Control Response
Metric 6: Randomized Allocation
Medium x 2
High x 1
Low	x 1
A negative control was used, but the study authors
did not specify the type of control used for the
methylene chloride test (i.e. dilution water, solvent
control, or filtered water not specified).
The negative control response was less than or equal
to 10 percent.
Randomization was not reported.
Domain 3: Exposure Characterization
Metric 7: Experimental System/Test Media Prepara- High
tion
Metric 8: Consistency of Exposure Administration	Low
Metric 9: Measurement of Test Substance Concentra- Medium x 1
tion
Metric 10: Exposure Duration and Frequency
High
Metric 11: Number of Exposure Groups/Spacing of Ex- Medium x 1
posure Levels
Metric 12: Testing at or Below Solubility Limit
High
X 2	2	The authors used teflon-lined screw caps with no air
space to minimize test substance loss.
X 1	3	Exposure concentrations were not provided, but all
test substances were diluted into a minimum of 5
exposure concentrations plus a control for each test.
2	Only nominal concentration were used, but measures
were taken to limit test chemical loss in the test
chambers.
X 2	2 These were static 48-hour tests which is sufficient for
determining acute effects in aquatic invertebrates.
2	There were 5 exposure concentrations plus controls
for each test substance, but concentrations used and
spacing of exposure levels was not reported.
X 1	1	Median 48-hour mortality was observed at concen-
trations below the water solubility limit.
Continued on next page . . .
38

-------
. continued from previous page
Study Citation: Abernethy, S.,Bobra, A. M.,Shiu, W. Y.,Wells, P. G.,Mackay, D.. 1986. ACUTE LETHAL TOXICITY OF HYDROCARBONS
AND CHLORINATED HYDROCARBONS TO TWO PLANKTONIC CRUSTACEANS THE KEY ROLE OF ORGANISM-WATER
PARTITIONING. Aquatic Toxicology
Data Type:	Acute (0-96 hour); Aquatic; Invertebrates
Hero ID:	1486051
Domain
Metric
Rating"!" MWF* Score
Comments'r

Domain 4: Test Organism
Metric
13:
Test Organism Characteristics
High
X
2
2
Daphnia magna, a well-known laboratory test organ-
ism, were used.
Metric
14:
Acclimitization and Pretreatment Conditions
Low
X
1
3
The study did not report whether organisms were
acclimatized.
Metric
15:
Number of Organisms and Replicates per
Group
Low
X
1
3
The number of organisms and replicates not indi-
cated.
Metric
16:
Adequacy of Test Conditions
Low
X
1
3
The temperatures (21-25"C) at which the daphnia
were kept were higher than the recommended range
of temperatures from OECD Test Guideline 202 (18-
22" C) and test temps varied greater than 1C. Load-
ing rate was unclear, but does not appear to have
affected results.
Domain 5: Outcome Assessment
Metric 17: Outcome Assessment Methodology	High	X 2	2 Median lethal concentration (LC50s) were reported.
Metric 18: Consistency of Outcome Assessment	Medium x 1	2 Incomplete details of test protocol reported, how-
ever, study indicated that results were corrected for
control mortality (which is less than 10 percent).
Domain 6: Confounding / Variable Control
Metric 19: Confounding Variables in Test Design and Medium x 2	4 Study did not provide enough information to allow
Procedures	a comParison f environmental conditions or other
non-treatment-related factors across study groups;
however, this is not likely to have significant impact
on study results as controls had <10 percent mor-
tality.
Metric 20: Outcomes Unrelated to Exposure	Medium x 1	2 Data on attrition for each study group was not re-
ported, but controls had < 10 percent mortality, so
unlikely to have significant impact on results.
Domain 7: Data Presentation and Analysis
Continued on next page . . .
39

-------
. continued from previous page
Study Citation: Abernethy, S.,Bobra, A. M.,Shiu, W. Y.,Wells, P. G.,Mackay, D.. 1986. ACUTE LETHAL TOXICITY OF HYDROCARBONS
AND CHLORINATED HYDROCARBONS TO TWO PLANKTONIC CRUSTACEANS THE KEY ROLE OF ORGANISM-WATER
PARTITIONING. Aquatic Toxicology
Data Type:	Acute (0-96 hour); Aquatic; Invertebrates
Hero ID:	1486051
Domain
Metric
Rating^
MWF*
Score
Comments'r
Metric 21:
Statistical Methods
Medium
X 1
2
The percent mortality at each concentration was cor-





rected for control mortality(always < 10 percent).





Nominal medial lethal concentrations (LC50 val-





ues), slope factors, and 95 percent confidence limits





were calculated from graphs by using Litchfield and





Wilcoxon (1948) methodology.
Metric 22:
Reporting of Data
Low
x 2
6
Data for exposure-related findings were not shown





for each study group, but results were described in





the test and or data were only reported for some





outcomes.
Metric 23:
Explanation of Unexpected Outcomes
High
x 1
1
No unexpected outcomes were reported.
Overall Quality Determination"'"

Medium

1.8

Extracted

Yes




* MWF = Metric Weighting Factor
t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value.
$ The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High.
Overall rating =
(Metric Score; X MWF;) / ]T\ MWF,
if any metric is Unacceptable
(round to the nearest tenth) otherwise
where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is
crossed out and an arrow points to the new rating.
U Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments.
40

-------
Study Citation: Dierickx, P. J.. 1993. Comparison between fish lethality data and the in vitro cytotoxicity of lipophilic solvents to cultured fish cells
in a two-compartment model. Chemosphere 27:1511-1518
Data Type:	Acute (0-96 hour); Aquatic; other fathead minnow cell-line; total protein content
Hero ID:	2803221
Domain
Metric
Rating1
MWF* Score
Comments' t
Domain 1: Test Substance
Metric 1:
Metric 2:
Metric 3:
Test Substance Identity
Test Substance Source
Test Substance Purity
High
Low
Low
X 2	2	Test substance was identified as methylene chloride.
X 1	3 DCM source not reported.
X 1	3 Purity was not indicated.
Domain 2: Test Design
Metric 4:
Metric 5:
Metric 6:
Negative Controls
Negative Control Response
Randomized Allocation
Medium	X 2	4 A control was used, but not much detail is given.
Low	X 1	3 Negative control response was not reported, in vitro
test.
Low	X 1	3 Not reported whether cells were allocated randomly-
in vitro test.
Domain 3: Exposure Characterization
Metric 7: Experimental System/Test Media Prepara- Low
tion
Metric 8: Consistency of Exposure Administration	High
Metric 9: Measurement of Test Substance Concentra- Low
tion
Metric 10: Exposure Duration and Frequency	Medium
Metric 11: Number of Exposure Groups/Spacing of Ex- Low
posure Levels
Metric 12: Testing at or Below Solubility Limit
x 2
x 1
x 1
x 2
x 1
Unacceptable x 1
DCM was solubilized in paraffin for this 24 hr in-
vitro test, but not clear that this reduced loss of
test substance from volatilization.
Exposure concentrations were not measured.
This was a 24-hour static test in fish cells, and ef-
fects were observed, but there is uncertainty whether
exposure duration was adequate.
Number of exposure groups and spacing were not
reported, but an EC50 was calculated.
Figure 1 in the paper indicates that the minimum
concentration tested and the EC50 are higher than
the water solubility for DCM, and there were no an-
alytical measurements of the test concentrations.
Domain 4: Test Organism
Metric 13: Test Organism Characteristics
Medium
x 2
Cultured fathead minnow cells from posterior of
anus, unclear whether acceptable for this type of in
vitro test.
Continued on next page . . .
41

-------
. . . continued from previous page
Study Citation: Dierickx, P. J.. 1993. Comparison between fish lethality data and the in vitro cytotoxicity of lipophilic solvents to cultured fish cells
in a two-compartment model. Chemosphere 27:1511-1518
Data Type:	Acute (0-96 hour); Aquatic; other fathead minnow cell-line; total protein content
Hero ID:	2803221
Domain
Metric
Rating1
MWF*
Score
Comments' t
Metric 14:
Acclimitization and Pretreatment Conditions
High
X
1
1

Metric 15:
Number of Organisms and Replicates per
Low
X
1
3
The number of replicates was not indicated, but






there were 6 x 105 cells/0.6 mL test well. It is un-






clear whether the cell concentration was adequate






for this test.
Metric 16:
Adequacy of Test Conditions
Low
X
1
3
Control response was not reported, and it is unclear






whether test conditions were adequate for this test.
Domain 5: Outcome Assessment





Metric 17:
Outcome Assessment Methodology
High
X
2
2
Total protein inhibition (EC50) was used to deter-






mine cytotoxicity.
Metric 18:
Consistency of Outcome Assessment
High
X
1
1

Domain 6: Confounding / Variable Control





Metric 19:
Confounding Variables in Test Design and
Low
X
2
6
The study did not provide enough information to

Procedures




allow a comparison of the environmental conditions





for each study group.
Metric 20:
Outcomes Unrelated to Exposure
Low
X
1
3
Cell attrition was not reported for DCM for each






test concentration.
Domain 7: Data Presentation and Analysis





Metric 21:
Statistical Methods
Medium
X
1
2
The statistical method used to derive the EC50 was






not reported in detail.
Metric 22:
Reporting of Data
Low
X
2
6
Data for exposure related findings were not reported






for each study group.
Metric 23:
Explanation of Unexpected Outcomes
Low
X
1
3
Authors did not find a linear correlation between






the published LC50 levels and the EC50 values from






this test and the reasoning for the discrepancy was






unclear.
Overall Quality Determination"'"
Unacceptable


4.0
Metric mean score**: 2.4.
Extracted

Yes





Continued on next page . . .
42

-------
. continued from previous page
Study Citation: Dierickx, P. J.. 1993. Comparison between fish lethality data and the in vitro cytotoxicity of lipophilic solvents to cultured fish cells
in a two-compartment model. Chemosphere 27:1511-1518
Data Type:	Acute (0-96 hour); Aquatic; other fathead minnow cell-line; total protein content
Hero ID:	2803221
Domain
Metric
Rating1
MWF* Score
Comments11

** Consistent with our Application of Systematic Review in TSCARisk Evaluations document, if a metric for a data source receives a score of Unacceptable (score = 4),
EPA will determine the study to be unacceptable. In this case, one of the metrics were rated as unacceptable. As such, the study is considered unacceptable and the score
is presented solely to increase transparency.
*	MWF = Metric Weighting Factor
t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value.
*	The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High.
4	if any metric is Unacceptable
Overall rating =
J]. (Metric Score; x MWF;) / J] . MWFj
(round to the nearest tenth) otherwise
where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed
out and an arrow points to the new rating,
ft Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments.
43

-------
Study Citation: Wu, S.,Zhang, H.,Yu, X.,Qiu, L.. 2014. Toxicological Responses of Chlorella vulgaris to Dichloromethane and Dichloroethane. Envi-
ronmental Engineering Science 31
Data Type:	Acute (0-96 hour); Aquatic; other Algae; MDA
Hero ID:	3493045
Domain
Metric
Rating1
MWF* Score
Comments' t
Domain 1: Test Substance
Metric 1:
Metric 2:
Metric 3:
Test Substance Identity
Test Substance Source
Test Substance Purity
High
Low
Low
X 2	2 Test substance was identified as dichloromethane.
X 1	3	Test substance source was not identified.
X 1	3 Purity not reported.
Domain 2: Test Design
Metric 4:
Metric 5:
Negative Controls
Negative Control Response
Metric 6: Randomized Allocation
High
High
Low
X 2	2 Use of controls was indicated.
X 1	1	Results reported as relative to controls and control
responses were shown in the figures for each end-
point measured.
X 1	3 Randomization not described.
Domain 3: Exposure Characterization
Metric 7: Experimental System/Test Media Prepara- Unacceptable x 2
tion
Metric 8: Consistency of Exposure Administration	High	x 1
Metric 9: Measurement of Test Substance Concentra- Unacceptable x 1
tion
Metric 10: Exposure Duration and Frequency	High
Metric 11: Number of Exposure Groups/Spacing of Ex-	High
posure Levels
Metric 12: Testing at or Below Solubility Limit	High
x 2
x 1
x 1
Test concentrations were not measured and no effort
to prevent loss through volatilization.
Volatility of the test substance was not taken into
account. Test concentrations were not measured to
account for potential loss through volatilization.
96-hour exposures prior to MDA assay.
There were 6 concentrations tested plus a control,
and spacing was adequate for dose-response analysis.
Test concentrations were below the water solubility
limit for methylene chloride.
Domain 4: Test Organism
Metric 13: Test Organism Characteristics	High
Metric 14: Acclimitization and Pretreatment Conditions Medium
X 2	2 Chlorella vulgaris is a well-known green algae
species.
X 1	2	Pretreatment conditions were not described, but this
was not expected to have an impact on the results,
as evidenced by control performance.
Continued on next page . ..
44

-------
. continued from previous page
Study Citation: Wu, S.,Zhang, H.,Yu, X.,Qiu, L.. 2014. Toxicological Responses of Chlorella vulgaris to Dichloromethane and Dichloroethane. Envi-
ronmental Engineering Science 31
Data Type:	Acute (0-96 hour); Aquatic; other Algae; MDA
Hero ID:	3493045
Domain
Metric
Rating1
MWF*
Score
Comments' t
Metric 15:
Number of Organisms and Replicates per
High
X
1
1
Initial concentration of 8 x 105 cells/mL with 3 repli-

Group




cates per test concentration.
Metric 16:
Adequacy of Test Conditions
High
X
1
1

Domain 5: Outcome Assessment





Metric 17:
Outcome Assessment Methodology
N/A


N/A
Referred to Zhang and Kirkham (1994) methodology






for analyzing MDA, relative to protein content.
Metric 18:
Consistency of Outcome Assessment
High
X
1
1
Results were reported relative to controls and pro-






tein content.
Domain 6: Confounding / Variable Control





Metric 19:
Confounding Variables in Test Design and
High
X
2
2


Procedures





Metric 20:
Outcomes Unrelated to Exposure
Medium
X
1
2
Authors did not report outcomes that affected con-






trols, but did not indicate that control MDA en-






zymes were within an acceptable range; however, the






authors report results relative to control values.
Domain 7: Data Presentation and Analysis





Metric 21:
Statistical Methods
High
X
1
1
Results were reported as the mean +/- standard er-






ror and used ANOVA to test for significance, with






less than 0.05 or 0.01 probability.
Metric 22:
Reporting of Data
High
X
2
2
Regression equation, correlation coefficient and sig-






nificance reported, along with NOEC, and figures






representing treatments relative to controls were re-






ported.
Metric 23:
Explanation of Unexpected Outcomes
High
X
1
1

Overall Quality Determination"'"
Unacceptable


4.0
Metric mean score**: 1.6.
Extracted

No





Continued on next page . . .
45

-------
. continued from previous page
Study Citation: Wu, S.,Zhang, H.,Yu, X.,Qiu, L.. 2014. Toxicological Responses of Chlorella vulgaris to Dichloromethane and Dichloroethane. Envi-
ronmental Engineering Science 31
Data Type:	Acute (0-96 hour); Aquatic; other Algae; MDA
Hero ID:	3493045
Domain
Metric
Rating1
MWF* Score
Comments11

** Consistent with our Application of Systematic Review in TSCARisk Evaluations document, if a metric for a data source receives a score of Unacceptable (score = 4),
EPA will determine the study to be unacceptable. In this case, two of the metrics were rated as unacceptable. As such, the study is considered unacceptable and the score
is presented solely to increase transparency.
*	MWF = Metric Weighting Factor
t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value.
*	The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High.
4	if any metric is Unacceptable
Overall rating =
J]. (Metric Score; x MWF;) / J] . MWFj
(round to the nearest tenth) otherwise
where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed
out and an arrow points to the new rating,
ft Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments.
46

-------
Study Citation: Wu, S.,Zhang, H.,Yu, X.,Qiu, L.. 2014. Toxicological Responses of Chlorella vulgaris to Dichloromethane and Dichloroethane. Envi-
ronmental Engineering Science 31
Data Type:	Acute (0-96 hour); Aquatic; other Algae; Chlorophyl a
Hero ID:	3493045
Domain
Metric
Rating1
MWF* Score
Comments' t
Domain 1: Test Substance
Metric 1:
Metric 2:
Metric 3:
Test Substance Identity
Test Substance Source
Test Substance Purity
High
Low
Low
X 2	2 Test material was identified as methylene chloride
X 1	3	Test substance source was not identified.
X 1	3 Purity was not reported.
Domain 2: Test Design
Metric 4:
Metric 5:
Metric 6:
Negative Controls
Negative Control Response
Randomized Allocation
High
High
Low
X 2	2 Use of controls indicated.
X 1	1	Percent inhibition values were reported relative to
controls and control responses indicated in figures.
X 1	3 Randomization not described.
Domain 3: Exposure Characterization
Metric 7: Experimental System/Test Media Prepara-
tion
Metric 8: Consistency of Exposure Administration
Metric 9: Measurement of Test Substance Concentra-
tion
Metric 10: Exposure Duration and Frequency
Metric 11: Number of Exposure Groups/Spacing of Ex-
posure Levels
Metric 12: Testing at or Below Solubility Limit
Unacceptable x 2
High	x 1
Unacceptable x 1
High
High
High
x 2
x 1
x 1
Test concentrations were not measured and no effort
to prevent loss through volatilization.
No analytical measurement of volatile test sub-
stance.
96-hour exposures prior to Chlorophyl a assay.
There were 6 concentrations tested plus a control,
and spacing was adequate for dose-response analysis.
Test concentrations were below the water solubility
limit for methylene chloride.
Domain 4: Test Organism
Metric 13: Test Organism Characteristics	High
Metric 14: Acclimitization and Pretreatment Conditions Medium
Metric 15: Number of Organisms and Replicates per High
Group
Metric 16: Adequacy of Test Conditions	High
x 2
x 1
x 1
x 1
Chlorella vulgaris is a well-known species of algae.
Pretreatment conditions were not described, but this
was not expected to have an impact on the results,
as evidenced by control performance.
Initial concentration was 8 x 105 cells/mL with 3
replicates per test concentration.
Continued on next page . . .
47

-------
. continued from previous page
Study Citation: Wu, S.,Zhang, H.,Yu, X.,Qiu, L.. 2014. Toxicological Responses of Chlorella vulgaris to Dichloromethane and Dichloroethane. Envi-
ronmental Engineering Science 31
Data Type:	Acute (0-96 hour); Aquatic; other Algae; Chlorophyl a
Hero ID:	3493045
Domain	Metric	Rating^	MWF* Score	Comments^
Domain 5: Outcome Assessment
Metric 17: Outcome Assessment Methodology
Metric 18: Consistency of Outcome Assessment
N/A
High
N/A Referred to Inskeep and Bloom (1985) method for
chlorophyll a analysis.
X 1	1	Results were reported relative to controls.
Domain 6: Confounding / Variable Control
Metric 19: Confounding Variables in Test Design and High	x 2 2
Procedures
Metric 20: Outcomes Unrelated to Exposure	Medium	x 1	2
Authors did not report outcomes that affected con-
trol outcomes, but did not indicate that control re-
sults were within acceptable ranges for the test as-
says. The authors do report results relative to con-
trol values.
Domain 7: Data Presentation and Analysis
Metric 21: Statistical Methods	High	X 1	1 Results were reported as the mean +/- standard er-
ror and used ANOVA to test for significance, with
less than 0.05 or 0.01 probability.
Metric 22: Reporting of Data	High	X 2	2 Regression equation, correlation coefficient and sig-
nificance reported, along with EC50.
Metric 23: Explanation of Unexpected Outcomes	High	x 1	1
Overall Quality Determination"'"
Unacceptable
4.0 Metric mean score**: 1.6.
Extracted
Yes
** Consistent with our Application of Systematic Review in TSCARisk Evaluations document, if a metric for a data source receives a score of Unacceptable (score = 4),
EPA will determine the study to be unacceptable. In this case, two of the metrics were rated as unacceptable. As such, the study is considered unacceptable and the score
is presented solely to increase transparency.
*	MWF = Metric Weighting Factor
t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value.
*	The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High.
Overall rating =
(Metric Score; x MWF;) / J] . MWFj
if any metric is Unacceptable
(round to the nearest tenth) otherwise
where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed
out and an arrow points to the new rating,
ft Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments.
48

-------
Study Citation: Wu, S.,Zhang, H.,Yu, X.,Qiu, L.. 2014. Toxicological Responses of Chlorella vulgaris to Dichloromethane and Dichloroethane. Envi-
ronmental Engineering Science 31
Data Type:	Acute (0-96 hour); Aquatic; other Algae; Protein Content
Hero ID:	3493045
Domain
Metric
Rating1
MWF* Score
Comments' t
Domain 1: Test Substance
Metric 1:
Metric 2:
Metric 3:
Test Substance Identity
Test Substance Source
Test Substance Purity
High
Low
Low
X 2	2 Test substance was identified as dichloromethane.
X 1	3	Test substance source was not identified.
X 1	3 Purity was not reported.
Domain 2: Test Design
Metric 4:
Metric 5:
Negative Controls
Negative Control Response
Metric 6: Randomized Allocation
High
High
Low
X 2	2 Use of controls was indicated.
X 1	1	Results reported as relative to controls and control
responses were shown in the figures for each end-
point measured.
X 1	3 Randomization not described.
Domain 3: Exposure Characterization
Metric 7: Experimental System/Test Media Prepara-
tion
Consistency of Exposure Administration
Measurement of Test Substance Concentra-
tion
Exposure Duration and Frequency
Number of Exposure Groups/Spacing of Ex-
posure Levels
Testing at or Below Solubility Limit
Metric 8:
Metric 9:
Metric 10:
Metric 11:
Metric 12:
Unacceptable x 2
High	x 1
Unacceptable x 1
High
High
High
x 2
x 1
x 1
No effort to prevent loss through volatilization.
Test concentrations were not measured and no effort
to prevent loss through volatilization.
96-hour exposures prior to protein content assay.
There were 6 concentrations tested plus a control,
and spacing was adequate for dose-response analysis.
Test concentrations were below the water solubility
limit for methylene chloride.
Domain 4: Test Organism
Metric 13: Test Organism Characteristics	High	x 2
Metric 14: Acclimitization and Pretreatment Conditions Medium	x 1
Metric 15: Number of Organisms and Replicates per High	x 1
Group
2	Chlorella vulgaris is a well-studied green algae
species.
2	Pretreatment conditions were not described, but this
was not expected to have an impact on the results,
as evidenced by control performance.
1	Initial concentratios were 8 x 105 cells/mL with 3
replicates per test concentration.
Continued on next page . ..
49

-------
. continued from previous page
Study Citation: Wu, S.,Zhang, H.,Yu, X.,Qiu, L.. 2014. Toxicological Responses of Chlorella vulgaris to Dichloromethane and Dichloroethane. Envi-
ronmental Engineering Science 31
Data Type: Acute (0-96 hour); Aquatic; other Algae; Protein Content
Hero ID: 3493045
Domain Metric
Rating1
MWF*
Score
Comments' t
Metric 16: Adequacy of Test Conditions
High
X 1
1

Domain 5: Outcome Assessment
Metric 17: Outcome Assessment Methodology
Metric 18: Consistency of Outcome Assessment
High
High
x 2
x 1
2
1
Protein content was determined using a BCA protein
assay.
Results were reported relative to controls.
Domain 6: Confounding / Variable Control
Metric 19: Confounding Variables in Test Design and
Procedures
Metric 20: Outcomes Unrelated to Exposure
High
Medium
x 2
x 1
2
2
Authors did not report outcomes that affected con-
trols, and did indicate whether control protein con-
tent was within an acceptable range; however, the
authors reported assay results relative to control val-
ues.
Domain 7: Data Presentation and Analysis
Metric 21: Statistical Methods
Metric 22: Reporting of Data
Metric 23: Explanation of Unexpected Outcomes
High
High
High
x 1
x 2
x 1
1
2
1
Regression equation, correlation coefficient and sig-
nificance reported, along with NOEC, and figures
representing treatments relative to controls were re-
ported.
Overall Quality Determination"'"
Unacceptable

4.0
Metric mean score**: 1.5.
Extracted
No



Continued on next page . . .
50

-------
. continued from previous page
Study Citation: Wu, S.,Zhang, H.,Yu, X.,Qiu, L.. 2014. Toxicological Responses of Chlorella vulgaris to Dichloromethane and Dichloroethane. Envi-
ronmental Engineering Science 31
Data Type:	Acute (0-96 hour); Aquatic; other Algae; Protein Content
Hero ID:	3493045
Domain
Metric
Rating1
MWF* Score
Comments11

** Consistent with our Application of Systematic Review in TSCARisk Evaluations document, if a metric for a data source receives a score of Unacceptable (score = 4),
EPA will determine the study to be unacceptable. In this case, two of the metrics were rated as unacceptable. As such, the study is considered unacceptable and the score
is presented solely to increase transparency.
*	MWF = Metric Weighting Factor
t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value.
*	The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High.
4	if any metric is Unacceptable
Overall rating =
J]. (Metric Score; x MWF;) / J] . MWFj
(round to the nearest tenth) otherwise
where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed
out and an arrow points to the new rating,
ft Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments.
51

-------
Study Citation: Wu, S.,Zhang, H.,Yu, X.,Qiu, L.. 2014. Toxicological Responses of Chlorella vulgaris to Dichloromethane and Dichloroethane. Envi-
ronmental Engineering Science 31
Data Type:	Acute (0-96 hour); Aquatic; other Algae; CAT and SOD
Hero ID:	3493045
Domain
Metric
Rating1
MWF* Score
Comments' t
Domain 1: Test Substance
Metric 1:
Metric 2:
Metric 3:
Test Substance Identity
Test Substance Source
Test Substance Purity
High
Low
Low
X 2	2 Test substance was identified as dichloromethane.
X 1	3	Test substance source was not identified.
X 1	3 Purity was not reported
Domain 2: Test Design
Metric 4:
Metric 5:
Negative Controls
Negative Control Response
Metric 6: Randomized Allocation
High
High
Low
X 2	2 Controls were used for the enzyme tests.
X 1	1	Results reported as relative to controls and control
responses were shown in the figures for each end-
point measured.
X 1	3 Randomization not described.
Domain 3: Exposure Characterization
Metric 7: Experimental System/Test Media Prepara- Unacceptable x 2
tion
Metric 8: Consistency of Exposure Administration	High	x 1
Metric 9: Measurement of Test Substance Concentra- Unacceptable x 1
tion
Metric 10: Exposure Duration and Frequency	High
Metric 11: Number of Exposure Groups/Spacing of Ex-	High
posure Levels
Metric 12: Testing at or Below Solubility Limit	High
x 2
x 1
x 1
Volatility of the test substance was not taken into
account.
Volatility of the test substance was not taken into
account. Test concentrations were not measured to
account for potential loss through volatilization.
96-hour exposures prior to the enzyme assays.
There were 6 concentrations tested plus a control,
and spacing was adequate for dose-response analysis.
Testing was below the water solubility limit.
Domain 4: Test Organism
Metric 13: Test Organism Characteristics	High
Metric 14: Acclimitization and Pretreatment Conditions Medium
Metric 15: Number of Organisms and Replicates per High
Group
X 2	2 Chlorella vulgaris is a well-studied algae species.
X 1	2	Pretreatment conditions were not described, but this
was not expected to have an impact on the results,
as evidenced by control performance.
X 1	1 Initial concentration of 8 x 105 cells/mL with 3 repli-
cates per test concentration.
Continued on next page . . .
52

-------
. continued from previous page
Study Citation: Wu, S.,Zhang, H.,Yu, X.,Qiu, L.. 2014. Toxicological Responses of Chlorella vulgaris to Dichloromethane and Dichloroethane. Envi-
ronmental Engineering Science 31
Data Type:	Acute (0-96 hour); Aquatic; other Algae; CAT and SOD
Hero ID:	3493045
Domain Metric
Rating1
MWF*
Score
Comments' t
Metric 16: Adequacy of Test Conditions
High
X
1
1

Domain 5: Outcome Assessment





Metric 17: Outcome Assessment Methodology
High
X
2
2
Catalase activity was assessed as the amount of





H202 degraded per minute at 25C. and SOD was





the amount of enzyme inhibiting 50 percent of a wa-





ter soluble tetrazolium (indicator of superoxide rad-





icals).
Metric 18: Consistency of Outcome Assessment
High
X
1
1
Results were reported relative to controls.
Domain 6: Confounding / Variable Control





Metric 19: Confounding Variables in Test Design and
High
X
2
2

Procedures





Metric 20: Outcomes Unrelated to Exposure
Medium
X
1
2
Authors did not report outcomes that affected con-





trols, and did not indicate that control enzymes were





within an acceptable range; however, the authors re-





port results relative to control values.
Domain 7: Data Presentation and Analysis





Metric 21: Statistical Methods
High
X
1
1
Results were reported as the mean +/- standard er-





ror and used ANOVA to test for significance, with





less than 0.05 or 0.01 probability.
Metric 22: Reporting of Data
High
X
2
2
Regression equation, correlation coefficient and sig-





nificance reported, along with NOEC, and figures





representing treatments relative to controls were re-





ported.
Metric 23: Explanation of Unexpected Outcomes
High
X
1
1

Overall Quality Determination"'"
Unacceptable


4.0
Metric mean score**: 1.5.
Extracted
No




Continued on next page . . .
53

-------
. continued from previous page
Study Citation: Wu, S.,Zhang, H.,Yu, X.,Qiu, L.. 2014. Toxicological Responses of Chlorella vulgaris to Dichloromethane and Dichloroethane. Envi-
ronmental Engineering Science 31
Data Type:	Acute (0-96 hour); Aquatic; other Algae; CAT and SOD
Hero ID:	3493045
Domain
Metric
Rating1
MWF* Score
Comments11

** Consistent with our Application of Systematic Review in TSCARisk Evaluations document, if a metric for a data source receives a score of Unacceptable (score = 4),
EPA will determine the study to be unacceptable. In this case, two of the metrics were rated as unacceptable. As such, the study is considered unacceptable and the score
is presented solely to increase transparency.
*	MWF = Metric Weighting Factor
t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value.
*	The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High.
4	if any metric is Unacceptable
Overall rating =
J]. (Metric Score; x MWF;) / J] . MWFj
(round to the nearest tenth) otherwise
where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed
out and an arrow points to the new rating,
ft Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments.
54

-------
Study Citation: Wu, S.,Zhang, H.,Yu, X.,Qiu, L.. 2014. Toxicological Responses of Chlorella vulgaris to Dichloromethane and Dichloroethane. Envi-
ronmental Engineering Science 31
Data Type:	Acute (0-96 hour); Aquatic; other Algae; Growth
Hero ID:	3493045
Domain	Metric	Rating^ MWF* Score	Comments^
Domain 1: Test Substance
Metric 1:
Metric 2:
Metric 3:
Test Substance Identity
Test Substance Source
Test Substance Purity
High
Low
Low
X 2	2 Test substance was identified as dichloromethane.
X 1	3	Test substance source was not identified
X 1	3 Purity was not reported
Domain 2: Test Design
Metric 4:
Metric 5:
Negative Controls
Negative Control Response
Metric 6: Randomized Allocation
High
High
Low
X 2	2 A negative control was reported.
X 1	1	Results were reported as relative to controls and con-
trol responses were shown in the figures for each end-
point measured.
X 1	3 Randomization was not reported.
Domain 3: Exposure Characterization
Metric 7: Experimental System/Test Media Prepara- Unacceptable x 2
tion
Metric 8: Consistency of Exposure Administration	High	x 1
Metric 9: Measurement of Test Substance Concentra- Unacceptable x 1
tion
Metric 10: Exposure Duration and Frequency	High
Metric 11: Number of Exposure Groups/Spacing of Ex- High
posure Levels
Metric 12: Testing at or Below Solubility Limit	High
x 2
x 1
x 1
Volatility of the test substance was not taken into
account. Capping of test vessels and no headspace
needed to reduce volatilization of DCM.
Test concentrations were not measured to account
for potential loss through volatilization.
Algae were exposed to methylene chloride for 96-
hours to assess growth inhibition.
There were 6 concentrations tested plus a control,
and spacing was adequate for dose-response analysis.
Test concentrations were below the water solubility
limit for methylene chloride.
Domain 4: Test Organism
Metric 13: Test Organism Characteristics	High
Metric 14: Acclimitization and Pretreatment Conditions Medium
X 2	2 .Chlorella vulgaris is a well-known green algae
species.
X 1	2	Pretreatment conditions were not described, but this
was not expected to have an impact on the results,
as evidenced by control performance.
Continued on next page . ..
55

-------
. continued from previous page
Study Citation: Wu, S.,Zhang, H.,Yu, X.,Qiu, L.. 2014. Toxicological Responses of Chlorella vulgaris to Dichloromethane and Dichloroethane. Envi-
ronmental Engineering Science 31
Data Type:	Acute (0-96 hour); Aquatic; other Algae; Growth
Hero ID:	3493045
Domain
Metric
Rating1
MWF*
Score
Comments' t
Metric 15:
Number of Organisms and Replicates per
High
X
1
1
Initial concentration of 8 x 105 cells/mL with 3 repli-

Group




cates per test concentration.
Metric 16:
Adequacy of Test Conditions
High
X
1
1

Domain 5: Outcome Assessment





Metric 17:
Outcome Assessment Methodology
High
X
2
2

Metric 18:
Consistency of Outcome Assessment
High
X
1
1
Results were reported relative to controls and pro-






tein content.
Domain 6: Confounding / Variable Control





Metric 19:
Confounding Variables in Test Design and
High
X
2
2


Procedures





Metric 20:
Outcomes Unrelated to Exposure
High
X
1
1
Authors did not report outcomes that affected con-






trols, but did not indicate that control growth rates






were within an acceptable range; however, the au-






thors reported growth rate for treatment popula-






tions relative to controls.
Domain 7: Data Presentation and Analysis





Metric 21:
Statistical Methods
High
X
1
1
Results were reported as the mean +/- standard er-






ror and used ANOVA to test for significance, with






less than 0.05 or 0.01 probability.
Metric 22:
Reporting of Data
High
X
2
2
Regression equation, correlation coefficient and sig-






nificance reported, along with EC50 and NOEC, and






figures representing treatments relative to controls






were reported.
Metric 23:
Explanation of Unexpected Outcomes
High
X
1
1

Overall Quality Determination"'"
Unacceptable


4.0
Metric mean score**: 1.5.
Extracted

No





Continued on next page . . .
56

-------
. continued from previous page
Study Citation: Wu, S.,Zhang, H.,Yu, X.,Qiu, L.. 2014. Toxicological Responses of Chlorella vulgaris to Dichloromethane and Dichloroethane. Envi-
ronmental Engineering Science 31
Data Type:	Acute (0-96 hour); Aquatic; other Algae; Growth
Hero ID:	3493045
Domain
Metric
Rating1
MWF* Score
Comments11

** Consistent with our Application of Systematic Review in TSCARisk Evaluations document, if a metric for a data source receives a score of Unacceptable (score = 4),
EPA will determine the study to be unacceptable. In this case, two of the metrics were rated as unacceptable. As such, the study is considered unacceptable and the score
is presented solely to increase transparency.
*	MWF = Metric Weighting Factor
t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value.
*	The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High.
4	if any metric is Unacceptable
Overall rating =
J]. (Metric Score; x MWF;) / J] . MWFj
(round to the nearest tenth) otherwise
where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed
out and an arrow points to the new rating,
ft Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments.
57

-------
Study Citation: Steiman, R.,Seiglemurandi, F.,Guiraud, P.,Benoitguyod, J. L.. 1995. TESTING OF CHLORINATED SOLVENTS ON MICROFUNGI.
Environmental Toxicology and Water Quality 10:283-285
Data Type:	Acute (0-96 hour); Aquatic; other soil fungi
Hero ID:	3559784
Domain
Metric
Rating1
MWF* Score
Comments11
Domain 1: Test Substance
Metric 1:
Metric 2:
Metric 3:
Test Substance Identity
Test Substance Source
Test Substance Purity
High
High
Low
X 2	2 Test substance was identified as dichloromethane
(DCM).
X 1	1	Test substance was from a chemical company.
X 1	3 Not indicated
Domain 2: Test Design
Metric 4:
Metric 5:
Metric 6:
Negative Controls
Negative Control Response
Randomized Allocation
Unacceptable x 2	8
N/A	N/A
Low	x 1	3
No control.
No control used.
No randomization indicated
Domain 3: Exposure Characterization
Metric 7: Experimental System/Test Media Prepara-
tion
Metric 8: Consistency of Exposure Administration
Metric 9: Measurement of Test Substance Concentra-
tion
Metric 10: Exposure Duration and Frequency
Metric 11: Number of Exposure Groups/Spacing of Ex-
posure Levels
Metric 12: Testing at or Below Solubility Limit
High
High
Medium
High
N/A
N/A
x 2
x 1
x 1
x 2
2
N/A
N/A
A closed system was used to reduce volatilization.
Not measured, however, due to differences in volatil-
ity, the atmosphericconcentrations were not the
same for the different solvents tested: 2.40 g L-l
for DCM .
Each solvent had one exposure concentration deter-
mined by the atmospheric pressure of the test vessel.
Each solvent had one exposure concentration deter-
mined by the atmospheric pressure of the test vessel.
Domain 4: Test Organism
Metric 13: Test Organism Characteristics	High
Metric 14: Acclimitization and Pretreatment Conditions High
Metric 15: Number of Organisms and Replicates per N/A
Group
x 2
x 1
2
1
N/A
The sediment fungal strains were cultivated in petri
dishes on synthetic solid medium of Galzy and
Slonimski (I 957)(glucose 5 g L - 1).
Each solvent had one exposure concentration deter-
mined by the atmospheric pressure of the test vessel;
testing on sediment fungal species.
Continued on next page . . .
58

-------
. continued from previous page
Study Citation: Steiman, R.,Seiglemurandi, F.,Guiraud, P.,Benoitguyod, J. L.. 1995. TESTING OF CHLORINATED SOLVENTS ON MICROFUNGI.
Environmental Toxicology and Water Quality 10:283-285
Data Type:	Acute (0-96 hour); Aquatic; other soil fungi
Hero ID:	3559784
Domain Metric
Rating1
MWF*
Score
Comments11
Metric 16: Adequacy of Test Conditions
High
X
1
1

Domain 5: Outcome Assessment





Metric 17: Outcome Assessment Methodology
High
X
2
2
Mortality of all test species increased with time.
Metric 18: Consistency of Outcome Assessment
High
X
1
1

Domain 6: Confounding / Variable Control





Metric 19: Confounding Variables in Test Design and
N/A


N/A
No further information was provided to determine
Procedures




if non-treatment related differences between test




groups.
Metric 20: Outcomes Unrelated to Exposure
N/A


N/A
No further information was provided to determine



outcomes unrelated to exposures.
Domain 7: Data Presentation and Analysis





Metric 21: Statistical Methods
N/A


N/A
Not provided.
Metric 22: Reporting of Data
Low
X
2
6
Results were reported but not raw data.
Metric 23: Explanation of Unexpected Outcomes
N/A


N/A
Not provided.
Overall Quality Determination"'"
Unacceptable


4.0
Metric mean score**: 1.7.
Extracted
Yes




** Consistent with our Application of Systematic Review in TSCARisk Evaluations document, if a metric for a data source receives a score of Unacceptable (score = 4), EPA
will determine the study to be unacceptable. In this case, one of the metrics were rated as unacceptable. As such, the study is considered unacceptable and the score is
presented solely to increase transparency.
*	MWF = Metric Weighting Factor
t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value.
*	The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High.
Overall rating =
^ (Metric Score; x MWF;) / ^ MWFj
if any metric is Unacceptable
(round to the nearest tenth) otherwise
where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed
out and an arrow points to the new rating.
^ Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments.
59

-------
Study Citation: Dill, D. C.,Murphy, P. G.,Mayes, M. A.. 1987. TOXICITY OF METHYLENE-CHLORIDE TO LIFE STAGES OF THE FATHEAD
MINNOW, PIMEPHALES-PROMELAS RAFINESQUE. Bulletin of Environmental Contamination and Toxicology 39:869-876
Data Type:	Other; Aquatic; Fish
Hero ID:	3587456
Domain	Metric	Ratingf MWF* Score	Comments"
Domain 1: Test Substance
Metric 1: Test Substance Identity	High X 2	2 Test substance was identified as methylene chloride.
Metric 2: Test Substance Source	High X 1	1 Test substance was obtained from Burdick and Jack-
son Laboratories, Muskegon, Michigin.
Metric 3: Test Substance Purity	High X 1	1 Test substance purity was > 99.9 percent.
Domain 2: Test Design
Metric 4:
Negative Controls
High
X
2
2
Negative controls were used in the test.
Metric 5:
Negative Control Response
High
X
1
1
There were no mortalities in the control group.
Metric 6:
Randomized Allocation
Low
X
1
3
Randomization procedure was not explained for
adult minnows
Domain 3: Exposure Characterization
Metric 7: Experimental System/Test Media Prepara- Low x 2
tion
Metric 8: Consistency of Exposure Administration	High x 1
Metric 9: Measurement of Test Substance Concentra- Low x 1
tion
Metric 10: Exposure Duration and Frequency	High x 2
Metric 11: Number of Exposure Groups/Spacing of Ex- High x 1
posure Levels
Metric 12: Testing at or Below Solubility Limit	High x 1
Test vessels were covered to prevent evaporation, but
headspace not addressed.
The test concentrations were reported in terms of
nominal test concentrations despite volatility and
head space in test vessels.
Flow-through exposures for 8-days (196 hours); a 96-
hour LC50was calculated.
There were 6 nominal text concentrations ranging
from 96 mg/L to 1020 mg/L.
Test concentrations were below the water solubility
limit for methylene chloride.
Domain 4: Test Organism
Metric 13: Test Organism Characteristics	High x 2	2
Metric 14: Acclimitization and Pretreatment Conditions High x 1	1
Fathead minnow were lab-reared.
Acclimatization and pretreatment conditions ap-
peared appropriate for the test, as evidenced by the
control group.
Continued on next page . . .
60

-------
. . . continued from previous page
Study Citation: Dill, D. C.,Murphy, P. G.,Mayes, M. A.. 1987. TOXICITY OF METHYLENE-CHLORIDE TO LIFE STAGES OF THE FATHEAD
MINNOW, PIMEPHALES-PROMELAS RAFINESQUE. Bulletin of Environmental Contamination and Toxicology 39:869-876
Data Type:	Other; Aquatic; Fish
Hero ID:	3587456
Domain
Metric
Rating^

MWF*
Score
Comments^1
Metric 15:
Number of Organisms and Replicates per
High
X
1
1
Two replicates of 10 fish were exposed to each treat-

Group




ment and a control vessel.
Metric 16:
Adequacy of Test Conditions
High
X
1
1
Test conditions were adequate and there wer no






mortalities in the control group.
Domain 5: Outcome Assessment





Metric 17:
Outcome Assessment Methodology
High
X
2
2
Water quality parameters, mortality and fish behav-






ior were observed daily to calculate the LC50.
Metric 18:
Consistency of Outcome Assessment
High
X
1
1
LC50s were assessed for all treatment groups.
Domain 6: Confounding / Variable Control





Metric 19:
Confounding Variables in Test Design and
High
X
2
2
No confounding variables in test design or proce-

Procedures




dures were reported.
Metric 20:
Outcomes Unrelated to Exposure
High
X
1
1
No outcomes unrelated to exposures were reported.
Domain 7: Data Presentation and Analysis





Metric 21:
Statistical Methods
High
X
1
1
Binomial or Probit analysis was used to calculate






the 96-hour LC50.
Metric 22:
Reporting of Data
High
X
2
2
Mortality data were presented in a table in the






results and discussion section of the paper.
Metric 23:
Explanation of Unexpected Outcomes
High
X
1
1
Unexpected outcomes were not reported.
Overall Quality Determination"'"
High
1.2
Extracted
Yes
Continued on next page . . .
61

-------
. continued from previous page
Study Citation: Dill, D. C.,Murphy, P. G.,Mayes, M. A.. 1987. TOXICITY OF METHYLENE-CHLORIDE TO LIFE STAGES OF THE FATHEAD
MINNOW, PIMEPHALES-PROMELAS RAFINESQUE. Bulletin of Environmental Contamination and Toxicology 39:869-876
Data Type:	Other; Aquatic; Fish
Hero ID:	3587456
Domain
Metric
Rating^
MWF*
Score
Comments^1

*	MWF = Metric Weighting Factor
t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value.
*	The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High.
if any metric is Unacceptable
Overall rating =
J]. (Metric Score; x MWF;) / ^ MWFj
(round to the nearest tenth) otherwise
where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is
crossed out and an arrow points to the new rating.
^ Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments.
62

-------
Study Citation: Thiebaud, H.,Merlin, G.,Capovilla, M. P.,Blake, G.. 1994. Fate of a volatile chlorinated solvent in indoor aquatic microcosms: sublethal
and static exposure to [14C]dichloromethane. Groupe pour l'Etude du Devenir de X" nobiotiques dans PEnvironnement (GEDEXE).
Ecotoxicology and Environmental Safety 28:71-81
Data Type:	Other; Aquatic; other mesocosm BCF
Hero ID:	3588425
Domain	Metric	Rating^ MWF* Score	Comments"^
Domain 1: Test Substance
Metric 1:
Test Substance Identity
High
X
2
2
Test substance was identified.
Metric 2:
Test Substance Source
Low
X
1
3
Source not reported.
Metric 3:
Test Substance Purity
Low
X
1
3
Purity of methylene chloride was not provided.
Domain 2: Test Design






Metric 4:
Negative Controls
High
X
2
2
Negative controls were used.
Metric 5:
Negative Control Response
N/A


N/A
This is a fate study conducted in a mesocosm, not a






toxicity test.
Metric 6:
Randomized Allocation
N/A


N/A
This is a fate study conducted in a mesocosm, not a




toxicity test.
Domain 3: Exposure Characterization





Metric 7:
Experimental System/Test Media Prepara-
High
X
2
2


tion





Metric 8:
Consistency of Exposure Administration
High
X
1
1

Metric 9:
Measurement of Test Substance Concentra-
High
X
1
1


tion





Metric 10:
Exposure Duration and Frequency
High
X
2
2

Metric 11:
Number of Exposure Groups/Spacing of Ex-
N/A


N/A


posure Levels





Metric 12:
Testing at or Below Solubility Limit
N/A


N/A
This is a fate study conducted in a mesocosm, not a






toxicity test.
Domain 4: Test Organism






Metric 13:
Test Organism Characteristics
High
X
2
2

Metric 14:
Acclimitization and Pretreatment Conditions
Low
X
1
3
Not indicated,
Metric 15:
Number of Organisms and Replicates per
High
X
1
1


Group





Metric 16:
Adequacy of Test Conditions
High
X
1
1

Continued on next page . . .
63

-------
. continued from previous page
Study Citation: Thiebaud, H.,Merlin, G.,Capovilla, M. P.,Blake, G.. 1994. Fate of a volatile chlorinated solvent in indoor aquatic microcosms: sublethal
and static exposure to [14C]dichloromethane. Groupe pour l'Etude du Devenir de X" nobiotiques dans PEnvironnement (GEDEXE).
Ecotoxicology and Environmental Safety 28:71-81
Data Type: Other; Aquatic; other mesocosm BCF
Hero ID: 3588425
Domain Metric
Rating^
MWF*
Score
Comments^

Domain 5: Outcome Assessment
Metric 17: Outcome Assessment Methodology
Metric 18: Consistency of Outcome Assessment
Unacceptable
N/A
x 2
8
N/A
NO adverse outcome- This study analyzed the bioac-
cumulation/concentration factors of DCM.
No adverse outcome- This study analyzed the bioac-
cumulation/concentration factors of DCM.
Domain 6: Confounding / Variable Control
Metric 19: Confounding Variables in Test Design and
Procedures
Metric 20: Outcomes Unrelated to Exposure
N/A
N/A

N/A
N/A
This study analyzed the bioaccumulation/
concentration factors of DCM.
This study analyzed the bioaccumulation/
concentration factors of DCM.
Domain 7: Data Presentation and Analysis
Metric 21: Statistical Methods
Metric 22: Reporting of Data
Metric 23: Explanation of Unexpected Outcomes
High
High
N/A
x 1
x 2
1
2
N/A
This study analyzed the bioaccumulation/
concentration factors of DCM, not a toxicity test.
Overall Quality Determination"'"
Unacceptable

4.0
Metric mean score**: 1.5.
Extracted
No
** Consistent with our Application of Systematic Review in TSCARisk Evaluations document, if a metric for a data source receives a score of Unacceptable (score = 4),
EPA will determine the study to be unacceptable. In this case, one of the metrics were rated as unacceptable. As such, the study is considered unacceptable and the score
is presented solely to increase transparency.
*	MWF = Metric Weighting Factor
1 High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value.
*	The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High.
Overall rating =
^ (Metric Score; x MWF;) / J] . MWF^
if any metric is Unacceptable
(round to the nearest tenth) otherwise
where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed
out and an arrow points to the new rating.
^ Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments.
64

-------
Study Citation: Rayburn, J. R.,Fisher, W. S.. 1999. Developmental toxicity of copper chloride, methylene chloride, and 6-aminonicotinamide to
embryos of the grass shrimp Palaemonetes pugio. Environmental Toxicology and Chemistry 18:950-957
Data Type:	Chronic (>21 days); Aquatic; Invertebrates
Hero ID:	3589368
Domain	Metric	Rating^ MWF* Score	Comments^
Domain 1: Test Substance
Metric 1: Test Substance Identity	High	X 2	2 Test substance identified as methylene chloride.
Metric 2: Test Substance Source	High	X 1	1 Test substance source was a chemical supplier,
Fisher Scientific.
Metric 3: Test Substance Purity	High	X 1	1 Test substance purity was 99.9 percent.
Domain 2: Test Design
Metric 4:	Negative Controls
Metric 5:	Negative Control Response
Metric 6:	Randomized Allocation
High
High
Low
X 2	2 A seawater control was used.
X 1	1	The average control mortality was reported as 4.2
percent, with one developmental delay and no ab-
normalities.
X 1	3	No randomization described in the study report.
Domain 3: Exposure Characterization
Metric 7: Experimental System/Test Media Prepara- Unacceptable x 2
tion
Metric 8: Consistency of Exposure Administration	High	x 1
Metric 9: Measurement of Test Substance Concentra- Unacceptable x 1
tion
Metric 10: Exposure Duration and Frequency
High
Metric 11: Number of Exposure Groups/Spacing of Ex- High
posure Levels
Metric 12: Testing at or Below Solubility Limit	High
x 2
x 1
x 1
The static test was conducted in glass tissue culture
tubes, but its unclear whether these were capped,
so loss of methylene chloride from volatilization was
possible.
Test concentrations were nominal for this volatile
substance and the test was static; its unclear
whether the test vessels were capped to reduce
volatilization.
The embryo-larvae test was 12 days (embryos were
exposed from 3 days to 15 days) in this static test.
Five concentrations plus the seawater control were
tested with 24 embryos per test concentration.
Test concentrations were all below the water solu-
bility of methylene chloride.
Domain 4: Test Organism
Continued on next page . . .
65

-------
. . . continued from previous page
Study Citation: Rayburn, J. R.,Fisher, W. S.. 1999. Developmental toxicity of copper chloride, methylene chloride, and 6-aminonicotinamide to
embryos of the grass shrimp Palaemonetes pugio. Environmental Toxicology and Chemistry 18:950-957
Data Type:	Chronic (>21 days); Aquatic; Invertebrates
Hero ID:	3589368
Domain
Metric
Rating1
MWF*
Score
Comments' t
Metric 13:
Test Organism Characteristics
Medium
X
2
4
Grass shrimp were collected from an estuary of un-






known water quality and not cultured for laboratory






testing. However, average control morality was 4.2






percent and 1 developmental delay, with no abnor-






malities, so these appear acceptable.
Metric 14:
Acclimitization and Pretreatment Conditions
High
X
1
1
The grass shrimp were acclimated to the aquaria for






at least 2 weeks.
Metric 15:
Number of Organisms and Replicates per
High
X
1
1
There were 3 replicates of 24 shrimp embryos per

Group




test concentration.
Metric 16:
Adequacy of Test Conditions
High
X
1
1

Domain 5: Outcome Assessment





Metric 17:
Outcome Assessment Methodology
High
X
2
2
Shrimp embryos were examined daily for heartbeat






and morphological abnormalties, hatching, and de-






velopmental delays.
Metric 18:
Consistency of Outcome Assessment
High
X
1
1

Domain 6: Confounding / Variable Control





Metric 19:
Confounding Variables in Test Design and
High
X
2
2
Test tubes were incubated at 27C and rotated at

Procedures




60 rpm, Details for water parameters were sparse,





but control response indicates it was within normal






ranges.
Metric 20:
Outcomes Unrelated to Exposure
High
X
1
1
Control responses indicate no outcomes unrelated to






exposures.
Domain 7: Data Presentation and Analysis





Metric 21:
Statistical Methods
High
X
1
1
LC50s with confidence intervals calculated with






Litchfield-Wilcox probit analysis.
Metric 22:
Reporting of Data
High
X
2
2

Metric 23:
Explanation of Unexpected Outcomes
High
X
1
1

Overall Quality Determination"'"
Unacceptable -


4.0
Metric mean score**: 1.4.
Extracted

No





Continued on next page . . .
66

-------
. continued from previous page
Study Citation: Rayburn, J. R.,Fisher, W. S.. 1999. Developmental toxicity of copper chloride, methylene chloride, and 6-aminonicotinamide to
embryos of the grass shrimp Palaemonetes pugio. Environmental Toxicology and Chemistry 18:950-957
Data Type:	Chronic (>21 days); Aquatic; Invertebrates
Hero ID:	3589368
Domain
Metric
Rating1
MWF* Score
Comments11

** Consistent with our Application of Systematic Review in TSCARisk Evaluations document, if a metric for a data source receives a score of Unacceptable (score = 4),
EPA will determine the study to be unacceptable. In this case, two of the metrics were rated as unacceptable. As such, the study is considered unacceptable and the score
is presented solely to increase transparency.
*	MWF = Metric Weighting Factor
t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value.
*	The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High.
if any metric is Unacceptable
Overall rating =
J]. (Metric Score; x MWF;) / J] . MWFj
(round to the nearest tenth) otherwise
where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed
out and an arrow points to the new rating,
ft Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments.
67

-------
Study Citation:
Rayburn, J. R.,Fisher, W. S.. 1999. Developmental toxicity of copper chloride, methylene chloride, and 6-aminonicotinamide to

embryos of the grass shrimp Palaemonetes pugio. Environmental Toxicology and Chemistry 18:950-957
Data Type:
Acute (0-96 hour); Aquatic; Invertebrates




Hero ID:
3589368




Domain
Metric
Rating^
MWF*
Score
Comments11
Domain 1: Test Substance





Metric 1: Test Substance Identity
High
x 2
2
Test substance identified as methylene chloride.

Metric 2: Test Substance Source
High
x 1
1
Test substance source was a chemical supplier,





Fisher Scientific.

Metric 3: Test Substance Purity
High
x 1
1
Test substance purity was 99.9 percent.
Domain 2: Test Design
Metric 4:
Metric 5:
Metric 6:
Negative Controls
Negative Control Response
Randomized Allocation
Higll	X 2	2 A seawater control was used.
High	X 1	1	The average control mortality was reported as 4.2
percent, with one developmental delay and no ab-
normalities.
Low	X 1	3 No randomization described in the study report.
Domain 3: Exposure Characterization
Metric 7: Experimental System/Test Media Prepara- Unacceptable x 2
tion
Metric 8:
Metric 9:
Metric 10:
Metric 11:
Metric 12:
Consistency of Exposure Administration	High x 1
Measurement of Test Substance Concentra- Unacceptable x 1
tion
Exposure Duration and Frequency	High	x 2
Number of Exposure Groups/Spacing of Ex-	High	x 1
posure Levels
Testing at or Below Solubility Limit	High	x 1
The static test was conducted in glass tissue culture
tubes, but it is unclear whether the tubes were capped
and no further steps to reduce loss of methylene chloride
from volatilization.
Test concentrations were nominal for this volatile
substance and the test was static. The test ves-
sels are tissue culture tubes, but it is unclear
whether tubes were capped and airspace was not minimized,
so loss of methylene chloride from volatilization was possible..
The embryo-larvae test was 96-hours
Five concentrations plus the seawater control were
tested with 24 embryos per test concentration.
Test concentrations were all below the water solu-
bility of methylene chloride.
Domain 4: Test Organism
Continued on next page
68

-------
. . . continued from previous page
Study Citation: Rayburn, J. R.,Fisher, W. S.. 1999. Developmental toxicity of copper chloride, methylene chloride, and 6-aminonicotinamide to
embryos of the grass shrimp Palaemonetes pugio. Environmental Toxicology and Chemistry 18:950-957
Data Type:	Acute (0-96 hour); Aquatic; Invertebrates
Hero ID:	3589368
Domain
Metric
Rating"!" MWF* Score
Comments^1
Metric 13: Test Organism Characteristics
Medium x 2
Metric 14: Acclimitization and Pretreatment Conditions	High	x 1	1
Metric 15: Number of Organisms and Replicates per	High	x 1	1
Group
Metric 16: Adequacy of Test Conditions	High	x 1	1
Grass shrimp were collected from an estuary of un-
known water quality and not cultured for laboratory
testing. However, average control morality was 4.2
percent and 1 developmental delay, with no abnor-
malities, so these appear acceptable.
The grass shrimp were acclimated to the aquaria for
at least 2 weeks.
There were 3 replicates of 24 shrimp embryos per
test concentration.
Domain 5: Outcome Assessment
Metric 17: Outcome Assessment Methodology
Metric 18: Consistency of Outcome Assessment
High
High
x 2
x 1
Shrimp embryos were examined daily for heartbeat
and morphological abnormalties.
Domain 6: Confounding / Variable Control
Metric 19: Confounding Variables in Test Design and High
Procedures
Metric 20: Outcomes Unrelated to Exposure
High
x 2
x 1
Test tubes were incubated at 27C and rotated at
60 rpm, Details for water parameters were sparse,
but control response indicates it was within normal
ranges.
Control responses indicate no outcomes unrelated to
exposures.
Domain 7: Data Presentation and Analysis
Metric 21: Statistical Methods
High
X
1
1
LC50s with confidence intervals calculated with





Litchfield-Wilcox probit analysis.
Metric 22: Reporting of Data
High
X
2
2

Metric 23: Explanation of Unexpected Outcomes
High
X
1
1

Overall Quality Determination"'"
Unacceptable
>
4.0
Metric mean score**: 1.3.
Continued on next page . . .
69

-------
. continued from previous page
Study Citation: Rayburn, J. R.,Fisher, W. S.. 1999. Developmental toxicity of copper chloride, methylene chloride, and 6-aminonicotinamide to
embryos of the grass shrimp Palaemonetes pugio. Environmental Toxicology and Chemistry 18:950-957
Data Type:	Acute (0-96 hour); Aquatic; Invertebrates
Hero ID:	3589368
Domain
Metric
Rating^
MWF* Score
Comments^1

Extracted

No



*	MWF = Metric Weighting Factor
t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value.
*	The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High.
Overall rating =
^ (Metric Score; x MWF;) / ^ MWFj
if any metric is Unacceptable
(round to the nearest tenth) otherwise
where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is
crossed out and an arrow points to the new rating.
^ Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments.
70

-------
Study Citation:	Birge, W. J.,Black, J. A.,Kuehne, R. A.. 1980. Effects of Organic Compounds on Amphibian Reproduction.
Data Type:	Acute (0-96 hour); Aquatic; other Amphibians
Hero ID:	3616521
Domain	Metric	Rating^ MWF* Score	Comments"^
Domain 1: Test Substance
Metric 1
Metric 2
Metric 3
Test Substance Identity
Test Substance Source
Test Substance Purity
High	X 2	2 Test substance identified as methylene chloride.
Low	X 1	3 Test substance source not indicated.
Medium X 1	2 All test substances were analytical or spectrophoto-
metric grade, but specific purity for methylene chlo-
ride was not indicated.
Domain 2: Test Design
Metric 4:
Negative Controls
High
X
2
2
Negative controls were 50 to 130 eggs.
Metric 5:
Negative Control Response
High
X
1
1
The control embryo/larvae survival ranged from 82
to 98 percent.
Metric 6:
Randomized Allocation
Low
X
1
3
Randomized allocation was not reported.
Domain 3: Exposure Characterization
Metric 7: Experimental System/Test Media Prepara- High
tion
Metric 8: Consistency of Exposure Administration	High
Metric 9: Measurement of Test Substance Concentra- High
tion
Metric 10: Exposure Duration and Frequency
Metric 12: Testing at or Below Solubility Limit
High
Metric 11: Number of Exposure Groups/Spacing of Ex- High
posure Levels
High
x 2
x 1
x 1
x 2
x 1
x 1
Flow-through test in a closed chamber without head
space to minimize volatilization, and daily analysis
of test substance concentrations during testing.
Gas chromatography was used to measure concen-
trations of methylene chloride in test vessels on a
daily basis.
Eggs and larvae were exposed up to 4 days post-
hatch using flow-through exposure concentrations.
Appropriate for embryo-larval EC50 and LC50 de-
termination.
Methylene chloride was tested at 5 concentrations
per species with appropriate spacing of exposure
concentrations.
Exposure concentrations tested were below methy-
lene chloride's water solubility limit.
Domain 4: Test Organism
Metric 13: Test Organism Characteristics	High	X 2	2 Amphibians obtained were appropriate for the
embryo-larval test.
Continued on next page . . .
71

-------
. continued from previous page
Study Citation: Birge, W. J.,Black, J. A.,Kuehne, R. A.. 1980. Effects of Organic Compounds on Amphibian Reproduction.
Data Type:	Acute (0-96 hour); Aquatic; other Amphibians
Hero ID:	3616521
Domain
Metric
Ratingt MWF* Score
Commentstt
Metric 14: Acclimitization and Pretreatment Conditions High x 1
Metric 15: Number of Organisms and Replicates per High x 1
Group
Metric 16: Adequacy of Test Conditions
Medium x 1
Fertilized eggs were either freshly obtained from the
local fish hatchery, or by ovulation induction and
fertilization of fresh eggs from frogs in the labora-
tory. All testing was initiated within 30 minutes of
fertilization.
Study indicated single replicates of 50 to 130 eggs
were used per methylene chloride exposure concen-
tration for each of three amphibian species.
There were up to 130 eggs per 500 mL test chamber.
Control survival was within normal limits, so this
appears acceptable.
Domain 5: Outcome Assessment
Metric 17: Outcome Assessment Methodology
Metric 18: Consistency of Outcome Assessment
High	X 2	2 Control-adjusted embryo-larval survival with daily
examination and removal of dead embryos/larvae.
High	X 1	1 LC50, LC10 and LCI values were adjusted for con-
trol mortality and teratogenicity.
Domain 6: Confounding / Variable Control
Metric 19: Confounding Variables in Test Design and High x 2 2
Procedures
Metric 20: Outcomes Unrelated to Exposure	Medium x 1	2
Control survival >80 percent and environmental
conditions were within acceptable ranges.
Control mortality ranged from 2-18 percent, how-
ever detailed control data was not provided.
Domain 7: Data Presentation and Analysis
Metric 21: Statistical Methods
Metric 22: Reporting of Data
Metric 23: Explanation of Unexpected Outcomes
Medium x 1
Medium x 2
High x 1
Finney's probit analysis was used to obtain LC50,
LC10 and LCI values with 95 percent confidence
limits; percent survival and mortality endpoints
were corrected for control mortality, but control data
was not reported.
Percent hatchability and survival of larvae reported
in table format for each test concentration, in ad-
dition to LC50, LC10 and LCI. Detailed control
mortality data was not reported.
Overall Quality Determination"'"
High
1.3
Continued on next page
72

-------
. continued from previous page
Study Citation: Birge, W. J.,Black, J. A.,Kuehne, R. A.. 1980. Effects of Organic Compounds on Amphibian Reproduction.
Data Type:	Acute (0-96 hour); Aquatic; other Amphibians
Hero ID:	3616521
Domain
Metric
Ratingt MWF* Score
Commentstt
Extracted
Yes
* MWF = Metric Weighting Factor
t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value.
The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High.
Overall rating =
(Metric Score; X MWF;) / J] . MWF,
if any metric is Unacceptable
(round to the nearest tenth) otherwise
where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is
crossed out and an arrow points to the new rating.
It Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments.
73

-------
Study Citation: Ando, T.,Otsuka, S.,Nishiyama, M.,Senoo, K.,Watanabe, M. M.,Matsumoto, S.. 2003. Toxic Effects of Dichioromethane and
Trichloroethylene on the Growth of Planktonic Green Algae, Chlorella vulgaris NIES227, Selenastrum capricornutum NIES35, and
Volvulina steinii NIES545. 18:43-46
Data Type:	Other; Aquatic; other Algae; Chlorophyll a
Hero ID:	3617103
Domain	Metric	Rating"!" MWF* Score	Comments"*1"!"
Domain 1: Test Substance
Metric 1:
Test Substance Identity
High
X
2
2
Test substance was identified as dichioromethane.
Metric 2:
Test Substance Source
Low
X
1
3
Test substance source not specified
Metric 3:
Test Substance Purity
Low
X
1
3
Purity was not specified
Domain 2: Test Design
Metric 4:
Negative Controls
High
X
2
2
Controls were included.
Metric 5:
Negative Control Response
High
X
1
1
Control responses were reported in figures
Metric 6:
Randomized Allocation
Low
X
1
3
Randomized allocation not indicated.
Domain 3: Exposure Characterization
Metric 7: Experimental System/Test Media Prepara- Low	x 2
tion
Metric 8: Consistency of Exposure Administration	High x 1
Metric 9: Measurement of Test Substance Concentra- Low	x 1
tion
Metric 10: Exposure Duration and Frequency	High x 2
Metric 11: Number of Exposure Groups/Spacing of Ex- High x 1
posure Levels
Metric 12: Testing at or Below Solubility Limit	High x 1
The vessels were covered, according to methods of
Arensberg et al., (1995) that authors referred to,
however headspace remained allowing for volatiliza-
tion of methylene chloride.
Test concentrations were reported in terms of nom-
inal concentrations and, according to methods of
Arensberg et al., (1995) loss of volatile solvent pos-
sible into test vessel headspace.
Treatment cultures were exposed for 10 days under
static conditions.
Including the controls, there were 7 exposure con-
centrations (nominal) that were spaced to detect a
concent rat ion-response relationship.
Testing was well below the water solubility limit of
methylene chloride.
Domain 4: Test Organism
Metric 13:
Test Organism Characteristics
High
x 2
Chlorella vulgaris and Selenestrum capricornutum
(now Raphidocelis subcapitata) are well studied
static algae species; Volvulina steiniis is a flagellar
algae.
Continued on next page
74

-------
. . . continued from previous page
Study Citation:
Data Type:
Hero ID:
Ando, T.,Otsuka, S.,Nishiyama, M.,Senoo, K.,Watanabe, M. M.,Matsumoto, S.. 2003. Toxic Effects of Dichloromethane and
Trichloroethylene on the Growth of Planktonic Green Algae, Chlorella vulgaris NIES227, Selenastrum capricornutum NIES35, and
Volvulina steinii NIES545. 18:43-46
Other; Aquatic; other Algae; Chlorophyll a
3617103
Domain
Metric
Rating"!" MWF* Score
Comments'r
Metric 14: Acclimitization and Pretreatment Conditions Low
Metric 15: Number of Organisms and Replicates per Medium x 1
Group
Metric 16: Adequacy of Test Conditions
High
X 1	3	Algae were incubated prior to addition of test mate-
rial, but details were not provided.
2 Five replicates at two different volumes (1 mL and 10
mL) for each algae species were tested, with initial
concentrations not specified.
X 1	1	Conditions were adequate as evidenced by control
vessel algal growth.
Domain 5: Outcome Assessment
Metric 17: Outcome Assessment Methodology
Metric 18: Consistency of Outcome Assessment
Low
High
x 2
x 1
Chlorophyll a light absorbance was reported for all
treatments and controls to measure growth, but is
not considered an accurate method for measuring
algal biomass.
Domain 6: Confounding / Variable Control
Metric 19: Confounding Variables in Test Design and High x 2 2
Procedures
Metric 20: Outcomes Unrelated to Exposure	High x 1	1
No inconsistencies in the test design and methodol-
ogy were reported.
Domain 7: Data Presentation and Analysis
Metric 21: Statistical Methods
Metric 22: Reporting of Data
Metric 23: Explanation of Unexpected Outcomes
Low
Low
High
x 1
x 2
x 1
Statistical analysis was performed to determine the
significance of differences between control and test
concentrations, but test methods were not pre-
sented.
Chlorophyll a absorbance as a measure of growth
was reported for all test concentrations, but an EC50
value was not calculated relative to the absorbance;
presented in figures only.
V. steinii test was repeated to confirm results.
Overall Quality Determination"'"	Medium	1.8
Extracted

Yes
Continued on next page . . .
75

-------
. continued from previous page
Study Citation: Ando, T.,Otsuka, S.,Nishiyama, M.,Senoo, K.,Watanabe, M. M.,Matsumoto, S.. 2003. Toxic Effects of Dichioromethane and
Trichioroethyiene on the Growth of Planktonic Green Algae, Chlorella vulgaris NIES227, Selenastrum capricornutum NIES35, and
Volvulina steinii NIES545. 18:43-46
Data Type:	Other; Aquatic; other Algae; Chlorophyll a
Hero ID:	3617103
Domain
Metric
Rating"'" MWF* Score
Comments1 r

* MWF = Metric Weighting Factor
t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value.
^ The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High.
if any metric is Unacceptable
Overall rating =
(Metric Score; X MWF;) / ]T\ MWF,
(round to the nearest tenth) otherwise
where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is
crossed out and an arrow points to the new rating,
tt Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments.
76

-------
Study Citation:	Wilson, J. E. H.. 1998. Developmental Arrest in Grass Shrimp Embryos Exposed to Selected Toxicants.
Data Type:	Aquatic; Invertebrates
Hero ID:	3617783
Domain	Metric	Rating^ MWF* Score	Comments"^
Domain 1: Test Substance
Metric 1:
Test Substance Identity
High
X
2
2
Test substance was identified as methylene chloride.
Metric 2:
Test Substance Source
Low
X
1
3
Test substance source was not indicated.
Metric 3:
Test Substance Purity
Low
X
1
3
Purity is not defined which meant the review was
unable to calculate the test concentrations in terms
of parts per million.
Domain 2: Test Design
Metric 4:
Negative Controls
High
X
2
2
Seawater controls were used.
Metric 5:
Negative Control Response
High
X
1
1
Results for negative controls reported
Metric 6:
Randomized Allocation
High
X
1
1
Stratified randomization method used
Domain 3: Exposure Characterization
Metric 7: Experimental System/Test Media Prepara- Low
tion
Metric 8: Consistency of Exposure Administration	High
Metric 9: Measurement of Test Substance Concentra- Low
tion
Metric 10: Exposure Duration and Frequency	High
Metric 11: Number of Exposure Groups/Spacing of Ex- High
posure Levels
Metric 12: Testing at or Below Solubility Limit	High
x 2
x 1
x 1
x 2
x 1
x 1
Flasks were covered parafilm to reduce evaporation,
and authors specified a static exposure regime, but
headspace in flasks allows volatilization of methylene
chloride.
Test was acute, but concentrations were not mea-
sured.
Exposure for 96-hours under static conditions.
Five exposure concentrations spaced exponentially.
Testing well below water solubility for methylene
chloride.
Domain 4: Test Organism
Metric 13: Test Organism Characteristics
High
Metric 14: Acclimitization and Pretreatment Conditions High
Metric 15: Number of Organisms and Replicates per Low
Group
X 2	2	Lab-cultured grass shrimp embryos well character-
ized.
X 1	1	Lab-cultured embryos for controls and treatment
groups appeared given same pre-treatment condi-
tions prior to testing.
X 1	3 30 embryos per flask, with two replications.
Continued on next page . . .
77

-------
. . . continued from previous page
Study Citation:
Wilson, J. E. H.. 1998. Developmental Arrest
in Grass Shrimp Embryos Exposed to Selected Toxicants.
Data Type:
Aquatic; Invertebrates

Hero ID:
3617783

Domain
Metric
Ratingt MWF* Score Commentstt

Metric 16: Adequacy of Test Conditions
High x 1 1
Domain 5: Outcome Assessment
Metric 17: Outcome Assessment Methodology
Metric 18: Consistency of Outcome Assessment
High
High
x 2
x 1
Embryos assessed for NOEC and LOEC for mortal-
ity, developmental delays/arrest and malformation
at 8 developmental stages through larval hatching.
Domain 6: Confounding / Variable Control





Metric 19:
Confounding Variables in Test Design and
High
X
2
2
None reported, and mortality/development abnor-

Procedures




malities were less than 10 percent in controls.
Metric 20:
Outcomes Unrelated to Exposure
High
X
1
1
Mortality and developmental abnormalities were less






than 10 percent for controls.
Domain 7: Data Presentation and Analysis





Metric 21:
Statistical Methods
Low
X
1
3
No statistical analysis were referenced, results were






reported in terms of NOAEC/LOAEC.
Metric 22:
Reporting of Data
Medium
X
2
4
Results for developmental delays, embryonic time






to develop, and mortality were reported for all test






chemicals.
Metric 23:
Explanation of Unexpected Outcomes
High
X
1
1

Overall Quality Determination"'"
High


1.5

Extracted

Yes





*	MWF = Metric Weighting Factor
t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value.
*	The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High.
Overall rating =
(Metric Score; x MWF;) / ^ MWFj
if any metric is Unacceptable
(round to the nearest tenth) otherwise
where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is
crossed out and an arrow points to the new rating.
Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments.
78

-------
Study Citation: Tsai, K. P.,Chen, C. Y.. 2007. An Algal Toxicity Database of Organic Toxicants Derived by a Closed-System Technique. Environmental
Toxicology and Chemistry 26:1931-1939
Data Type:	Acute (0-96 hour); Aquatic; Plants
Hero ID:	3617867
Domain	Metric	Rating^ MWF* Score	Comments^
Domain 1: Test Substance
Metric 1:
Test Substance Identity
High
X
2
2
Test substance was identified as methylene chloride.
Metric 2:
Test Substance Source
Low
X
1
3
A source was not provided.
Metric 3:
Test Substance Purity
High
X
1
1
The authors described the chemical purity as
"reagent grade"
Domain 2: Test Design
Metric 4: Negative Controls
Metric 5: Negative Control Response
Metric 6: Randomized Allocation
Medium X 2	4 Authors referred to a control, but additional details
were not reported.
Low	X 1	3 Negative Control response was not specifically re-
ported in the study, but was incorporated into the
calculation of the percent inhibition.
Low	X 1	3 Researchers did not report how organisms were al-
located to study groups
Domain 3: Exposure Characterization
Metric 7: Experimental System/Test Media Prepara-
tion
Metric 8: Consistency of Exposure Administration
Metric 9: Measurement of Test Substance Concentra-
tion
Metric 10: Exposure Duration and Frequency
Metric 12: Testing at or Below Solubility Limit
High
High
Medium
Metric 11: Number of Exposure Groups/Spacing of Ex- Low
posure Levels
Low
x 2
x 1
x 1
Medium x 2
x 1
x 1
Test systems were sealed with no headspace to elim-
inate volatilization.
Test concentrations were reported in terms of nom-
inal concentrations, but analytical confirmation of
the test concentrations was performed at the begin-
ning and end of the test and vessels were sealed with
no headspace.
The test duration was 48 hours, so results are not
comparable with 72 to 96 hours for algal testing.
The study report indicated that both a range finding
and definitive test were conducted but did not report
the test concentrations.
Test concentrations were not provided, but the LC50
is below the water solubility of DCM.
Domain 4: Test Organism
Continued on next page . . .
79

-------
. continued from previous page
Study Citation: Tsai, K. P.,Chen, C. Y.. 2007. An Algal Toxicity Database of Organic Toxicants Derived by a Closed-System Technique. Environmental
Toxicology and Chemistry 26:1931-1939
Data Type:	Acute (0-96 hour); Aquatic; Plants
Hero ID:	3617867
Domain	Metric	Rating^ MWF* Score	Comments^
Metric 13:
Test Organism Characteristics
High
X
2
2
Pseudokirchneriella subcapitata is a well-known al-






gal species.
Metric 14:
Acclimitization and Pretreatment Conditions
High
X
1
1

Metric 15:
Number of Organisms and Replicates per
High
X
1
1
15,000 algal cells/mL inoculum at test initiation,

Group




with 3 replicates.
Metric 16:
Adequacy of Test Conditions
High
X
1
1

Outcome Assessment





Metric 17:
Outcome Assessment Methodology
High
X
2
2
Outcome assessed at percent inhibition using an






electronic particle counter to evaluate cell density






at test initiation for controls and treatments.
Metric 18:
Consistency of Outcome Assessment
High
X
1
1
percent Inhibition was calculated relative to con-
trols.
Domain 6: Confounding / Variable Control
Metric 19: Confounding Variables in Test Design and High
Procedures
Metric 20: Outcomes Unrelated to Exposure	High
x 2
x 1
Authors conducted concentration checks where a
nominal test concentration was compared to a mea-
sured concentration from a vessel without algae.
The test was repeated if these concentrations dif-
fered greater than 6 percent,
percent inhibition was reported relative to control
concentrations.
Domain 7: Data Presentation and Analysis
Metric 21: Statistical Methods
Metric 22: Reporting of Data
Metric 23: Explanation of Unexpected Outcomes
High
Medium
High
X
X
X
1
2
1
1
4 Results did not include raw data.
1
Overall Quality Determination"'"
High


1.5
Extracted
Yes




Continued on next page . . .

-------
. . . continued from previous page
Study Citation:
Data Type:
Hero ID:
Tsai, K. P.,Chen, C. Y.. 2007. An Algal Toxicity Database of Organic Toxicants Derived by
Toxicology and Chemistry 26:1931-1939
Acute (0-96 hour); Aquatic; Plants
3617867
a Closed-System Technique. Environmental
Domain
Metric Rating"!" MWF* Score
Comments'^

*	MWF = Metric Weighting Factor
t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value.
*	The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High.
Overall rating =
^ (Metric Score; x MWF;) / ^ MWFj
if any metric is Unacceptable
(round to the nearest tenth) otherwise
where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is
crossed out and an arrow points to the new rating.
^ Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments.
81

-------
Study Citation:	Kramer, V. C.,Schnell, D. J.,Nickerson, K. W.. 1983. Relative Toxicity of Organic Solvents to Aedes aegypti Larvae. 42:285-287
Data Type:	Acute (0-96 hour); Aquatic; other Insect-larvae 4-hour LC50
Hero ID:	3661235
Domain	Metric	Rating^ MWF* Score	Comments"^
Domain 1: Test Substance
Metric 1:
Metric 2:
Test Substance Identity
Test Substance Source
Metric 3: Test Substance Purity
High	X 2	2 Test chemical was identified as methylene chloride.
Medium	X 1	2 Chemicals were described as " reagent grade" but no
further information was provided.
Medium	X 1	2 Chemicals were described as " reagent grade" but no
further information was provided.
Domain 2: Test Design
Metric 4:
Metric 5:
Metric 6:
Negative Controls
Negative Control Response
Randomized Allocation
High
High
N/A
X 2	2 Controls were used in this test.
X 1	1	There was no mortality observed in the controls.
N/A A single replicate of 10-20 larvae were transferred
from stock maintained in glass beakers/jars to 50
mL beakers for testing.
Domain 3: Exposure Characterization
Metric 7: Experimental System/Test Media Prepara- Low
tion
Metric 8: Consistency of Exposure Administration	Low
x 2
x 1
Metric 9: Measurement of Test Substance Concentra- Unacceptable x 1
tion
Metric 10: Exposure Duration and Frequency	Low	x 2
Metric 11: Number of Exposure Groups/Spacing of Ex- Unacceptable x 1
posure Levels
Metric 12: Testing at or Below Solubility Limit	N/A
6	No specific deficiencies were identified, but scarce
details were provided regarding the test preparation.
3	Exposure was described as a bioassay, which was re-
peated three times for each test chemical, but the
study report lacked sufficient details describing the
bioassay procedure.
4	Test concentrations were not provided, only end-
points were provided.
6	4 Hour duration were sufficient for the needs of the
authors, as they were looking to measure acute toxic-
ity. But this limited duration has low utility outside
of the results of this study.
4	Details about the exposure concentrations were un-
clear.
N/A Details about the exposure concentrations were un-
clear.
Domain 4: Test Organism
Metric 13: Test Organism Characteristics	High	x 2	2 Larva of Aedis aegypti (mosquitos).
Continued on next page . ..
82

-------
. continued from previous page
Study Citation: Kramer, V. C.,Schnell, D. J.,Nickerson, K. W.. 1983. Relative Toxicity of Organic Solvents to Aedes aegypti Larvae. 42:285-287
Data Type:	Acute (0-96 hour); Aquatic; other Insect-larvae 4-hour LC50
Hero ID:	3661235
Domain
Metric
Haling1
MWF*
Score
Commentstt
Metric 14:
Acclimitization and Pretreatment Conditions
Medium
X
1
2
No Acclimatization period reported, but embryos






obtained from research laboratory and hatched.






There were no mortalities in controls indicating ad-






equate pretreatment conditions.
Metric 15:
Number of Organisms and Replicates per
Low
X
1
3
A single replicate of 10-20 individuals per chemical,

Group




number not specified for DCM.
Metric 16:
Adequacy of Test Conditions
Medium
X
1
2
Individuals larvae were raised on dog food in baby






food jars prior to testing. Although there is a lack of






guideline for mosquito larvae maintenance, the con-






trols had no mortaility, indicating that conditions






were adequate for the test period.
Domain 5: Outcome Assessment





Metric 17:
Outcome Assessment Methodology
High
X
2
2

Metric 18:
Consistency of Outcome Assessment
Low
X
1
3
Details about the test procedure were not provided
Domain 6: Confounding / Variable Control





Metric 19:
Confounding Variables in Test Design and
Low
X
2
6
The authors did not provide enough information to

Procedures




allow for a comparison of the environmental condi-





tions or other non treatment related factors.
Metric 20:
Outcomes Unrelated to Exposure
High
X
1
1

Domain 7: Data Presentation and Analysis





Metric 21:
Statistical Methods
Unacceptable
X
1
4
Data were not provided, description of the statistical






analyses were not provided.
Metric 22:
Reporting of Data
Low
X
2
6
Only endpoint values (LC50s) were provided.
Metric 23:
Explanation of Unexpected Outcomes
High
X
1
1

Overall Quality Determination"'"
Unacceptable


4.0
Metric mean score**: 2.2.
Extracted

No





Continued on next page . . .
83

-------
. continued from previous page
Study Citation: Kramer, V. C.,Schnell, D. J.,Nickerson, K. W.. 1983. Relative Toxicity of Organic Solvents to Aedes aegypti Larvae. 42:285-287
Data Type:	Acute (0-96 hour); Aquatic; other Insect-larvae 4-hour LC50
Hero ID:	3661235
3661235
Domain
Metric
Ratingt	MWF* Score
Comments^
** Consistent with our Application of Systematic Review in TSCARisk Evaluations document, if a metric for a data source receives a score of Unacceptable (score = 4),
EPA will determine the study to be unacceptable. In this case, three of the metrics were rated as unacceptable. As such, the study is considered unacceptable and the
score is presented solely to increase transparency.
*	MWF = Metric Weighting Factor
t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value.
*	The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High.
where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed
out and an arrow points to the new rating,
ft Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments.
Overall rating
J]. (Metric Score; x MWF;) / ^ MWFj
o.i
if any metric is Unacceptable
(round to the nearest tenth) otherwise
84

-------
Study Citation:	Minnesota Mining & Mfg Co. 1979. 96-HOUR LC50 AQUATIC TEST ON FATHEAD MINNOWS WITH COVER LETTER.
Data Type:	Acute (0-96 hour); Aquatic; other Mixture 63 percent DCM; fathead minnow
Hero ID:	4213679
Domain	Metric	Rating^ MWF* Score	Comments
Domain 1: Test Substance
Metric 1: Test Substance Identity	Unacceptable	x2 8 Mixture with 63 percent methylene chloride;, and test results could not be
^	hvt/.	,,,, attributed solely to methylene chloride.
Metric 2: Test Substance Source	N/A	N/A
Metric 3: Test Substance Purity	N/A	N/A
Domain 2: Test Design



Metric 4:
Negative Controls
N/A
N/A
Metric 5:
Negative Control Response
N/A
N/A
Metric 6:
Randomized Allocation
N/A
N/A
Domain 3: Exposure Characterization


Metric 7:
Experimental System/Test Media Prepara-
N/A
N/A

tion


Metric 8:
Consistency of Exposure Administration
N/A
N/A
Metric 9:
Measurement of Test Substance Concentra-
N/A
N/A

tion


Metric 10:
Exposure Duration and Frequency
N/A
N/A
Metric 11:
Number of Exposure Groups/Spacing of Ex-
N/A
N/A

posure Levels


Metric 12:
Testing at or Below Solubility Limit
N/A
N/A
Domain 4: Test Organism



Metric 13:
Test Organism Characteristics
N/A
N/A
Metric 14:
Acclimitization and Pretreatment Conditions
N/A
N/A
Metric 15:
Number of Organisms and Replicates per
N/A
N/A

Group


Metric 16:
Adequacy of Test Conditions
N/A
N/A
Domain 5: Outcome Assessment


Metric 17:
Outcome Assessment Methodology
N/A
N/A
Metric 18:
Consistency of Outcome Assessment
N/A
N/A
Continued on next page . . .
85

-------
. continued from previous page
Study Citation: Minnesota Mining & Mfg Co. 1979. 96-HOUR LC50 AQUATIC TEST ON FATHEAD MINNOWS WITH COVER LETTER.
Data Type:	Acute (0-96 hour); Aquatic; other Mixture 63 percent DCM; fathead minnow
Hero ID:	4213679
Domain Metric
Haling1
MWF* Score Comments

Domain 6: Confounding / Variable Control
Metric 19: Confounding Variables in Test Design and
Procedures
Metric 20: Outcomes Unrelated to Exposure
N/A
N/A
N/A
N/A
Domain 7: Data Presentation and Analysis
Metric 21: Statistical Methods
Metric 22: Reporting of Data
Metric 23: Explanation of Unexpected Outcomes
N/A
N/A
N/A
N/A
N/A
N/A
Overall Quality Determination"'"
Unacceptable
4.0 Metric mean score**: 4.0.
Extracted
No


** Consistent with our Application of Systematic Review in TSCARisk Evaluations document, if a metric for a data source receives a score of Unacceptable (score = 4),
EPA will determine the study to be unacceptable. In this case, one of the metrics were rated as unacceptable. As such, the study is considered unacceptable and the score
is presented solely to increase transparency.
*	MWF = Metric Weighting Factor
( High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value.
*	The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High.
Overall rating =
^ (Metric Score; x MWF;) / J] . MWFj
if any metric is Unacceptable
(round to the nearest tenth) otherwise
where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed
out and an arrow points to the new rating.
86

-------
Study Citation: E I Dupont Denemours & Co Inc. 1987. FLOW-THROUGH ACUTE 96-HOUR LC50 OF METHYLENE CHLORIDE TO RAINBOW
TROUT (SANITIZED).
Data Type:	Acute (0-96 hour); Aquatic; Fish
Hero ID:	4213816
Domain	Metric	Rating^ MWF* Score	Comments^
Domain 1: Test Substance
Metric 1:
Test Substance Identity
High
X
2
2
Test substance identified by name and CAS.
Metric 2:
Test Substance Source
Low
X
1
3
Source not identified.
Metric 3:
Test Substance Purity
Low
X
1
3
Purity not reported.
Domain 2: Test Design
Metric 4: Negative Controls	Medium X 2	4 Negative control was used. A small amount of test
substance was detected in the control but it does not
seemed to have had an effect on the fish.
Metric 5: Negative Control Response	High	X 1	1 Control response was reported and had 0 percent
mortality.
Metric 6: Randomized Allocation	High	X 1	1 Ten juvenile rainbow trout were randomly placed in
the test vessels.
Domain 3: Exposure Characterization
Metric 7: Experimental System/Test Media Prepara- High x 2
tion
Metric 8: Consistency of Exposure Administration
High
Metric 9: Measurement of Test Substance Concentra- High
tion
Metric 10: Exposure Duration and Frequency
High
Metric 11: Number of Exposure Groups/Spacing of Ex- High
posure Levels
Metric 12: Testing at or Below Solubility Limit
High
x 1
x 1
x 2
x 1
x 1
2	Experimental system was described and appropri-
ate. Although it was not mentioned whether the
system was enclosed, the test was flow through and
test substance was renewed every 20 minutes. This
is important for a volatile chemical like methylene
chloride.
1	Exposures were administered consistently across
study groups.
1	Measurements of the test substance in each test con-
centration and in the control were taken every 24
hours.
2	The duration, 96 hours, and exposure frequency
(flow-through with renewal every 20 min) are rec-
ommended durations and frequency according to
OECD TG 203.
1	Authors reported 6 exposure groups, which meet the
minimum recommended in OECD TG 203, and spac-
ing was appropriate.
1	Test concentrations were well below the solubility of
methylene chloride.
Continued on next page . . .
87

-------
. . . continued from previous page
E I Dupont Denemours & Co Inc. 1987. FLOW-THROUGH ACUTE 96-HOUR LC50 OF METHYLENE CHLORIDE TO RAINBOW
TROUT (SANITIZED).
Acute (0-96 hour); Aquatic; Fish
4213816
Study Citation:
Data Type:
Hero ID:
Domain
Metric
Rating"'" MWF* Score
Comments^

Domain 4: Test Organism
Metric 13: Test Organism Characteristics
High
Metric 14: Acclimitization and Pretreatment Conditions High
x 2
x 1
x 1
Metric 15: Number of Organisms and Replicates per High
Group
Metric 16: Adequacy of Test Conditions	Medium x 1
2 Rainbow trout are a recommended species in OECD
TG 203. Fish were from Haskell lab stock.
1	Fish were held for about 94 days before the test,
which meets the min number of days to hold the
fish recommended in OECD TG 203. N treatment of
the fish for diseases was required during the holding
period.
1	Ten fish per test concentration were used. OECD
TG 203 recommends at least 7.
2	The 3.8 g fish/L loading may be acceptable given
that this is a flow-through tests (OECD TG 203 rec-
ommends 1 g fish/L for static tests but could be
higher for flow-through). Only minor uncertainties
about housing conditions, not likely to have a sub-
stantial impact on the results, given the 0 percent
mortality observed in the controls.
Domain 5: Outcome Assessment
Metric 17: Outcome Assessment Methodology
Metric 18: Consistency of Outcome Assessment
High	X 2	2 This test was able to derive an LC50.
High	X 1	1	Outcomes were assessed consistently across study
groups.
Domain 6: Confounding / Variable Control
Metric 19: Confounding Variables in Test Design and High x 2
Procedures
Metric 20: Outcomes Unrelated to Exposure
Medium x 1
2	There were no reported differences among the study
groups in environmental conditions or other factors
that could influence the outcome assessment.
2	Data on outcomes unrelated to exposure were not
reported for each study group. Only substantial dif-
ferences between groups was noted.
Domain 7: Data Presentation and Analysis
Metric 21: Statistical Methods
Metric 22: Reporting of Data
Low	X 1	3 The statistical method was not described clearly.
High	X 2	2 The results of the test were reported for each study
group in a table, and any effects besides mortality
were described in the text.
Continued on next page
88

-------
. . . continued from previous page
Study Citation: E I Dupont Denemours & Co Inc. 1987. FLOW-THROUGH ACUTE 96-HOUR LC50 OF METHYLENE CHLORIDE TO RAINBOW
TROUT (SANITIZED).
Data Type:	Acute (0-96 hour); Aquatic; Fish
Hero ID:	4213816
Domain	Metric	Rating^ MWF* Score	Comments^
Metric 23: Explanation of Unexpected Outcomes	High	X 1	1 Authors noted the small concentrations of test chem-
ical in the control, and hypothesized about the
source, but this does not seem to have affected the
results.
Overall Quality Determination"'"
High
1.3
Extracted
Yes


*	MWF = Metric Weighting Factor
t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value.
*	The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High.
4	if any metric is Unacceptable
Overall rating =
J]. (Metric Score; x MWF;) / J] . MWFj
(round to the nearest tenth) otherwise
where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is
crossed out and an arrow points to the new rating.
^ Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments.
89

-------
Study Citation: E I Dupont Denemours & Co Inc. 1987. DAPHNIA MAGNA STATIC ACUTE 48-HOUR EC50 OF METHYLENE CHLORIDE
(SANITIZED).
Data Type:	Acute (0-96 hour); Aquatic; Invertebrates
Hero ID:	4213817
Domain	Metric	Rating^ MWF* Score	Comments^
Domain 1: Test Substance
Metric 1:
Test Substance Identity
High
X
2
2
Chemical identified by name and CAS.
Metric 2:
Test Substance Source
Low
X
1
3
Test substance source was not reported.
Metric 3:
Test Substance Purity
Low
X
1
3
Test substance purity was not reported.
Domain 2: Test Design






Metric 4:
Negative Controls
High
X
2
2
Study included a control using water.
Metric 5:
Negative Control Response
High
X
1
1
Control response was reported and no immobiliza-
tion was observed in the control.
Metric 6:
Randomized Allocation
Low
X
1
3
Researchers did not report how organisms were al-
located to study groups.
Domain 3: Exposure Characterization





Metric 7:
Experimental System/Test Media Prepara-
Low
X
2
6
Authors did not report covering vessels and methy-

tion




lene chloride is a very volatile chemical. However,





they did measure concentrations at day 0 and day 2.
Contaminants were also noted, leaving uncertainties
about what was causing the observed toxicity.
Metric 8:
Consistency of Exposure Administration
High
X
1
1
Details of exposure administration were reported
and exposures were administered consistently across
study groups.
Metric 9:
Measurement of Test Substance Concentra-
High
X
1
1
Test concentrations were measured at day 0 and day

tion




2, and an average of those measurements was used





at each concentration level.
Metric 10:
Exposure Duration and Frequency
High
X
2
2
The test was a static 48-hour tests which is recom-
mended by OECD TG 202 for acute daphnia tests.
Metric 11:
Number of Exposure Groups/Spacing of Ex-
posure Levels
High
X
1
1
At least 5 test concentrations were used, and authors
were able to derive an EC50 with the concentrations
tested.
Metric 12:
Testing at or Below Solubility Limit
Medium
X
1
2
The test concentrations are far below the solubil-
ity limit for methylene chloride. However, the di-
vergence between nominal and measured concentra-
tions increases with concentration level, creating mi-
nor uncertainties.
Continued on next page . . .
90

-------
. continued from previous page
Study Citation:
E I Dupont Denemours & Co Inc. 1987.
DAPHNIA MAGNA STATIC ACUTE 48-HOUR EC50 OF METHYLENE CHLORIDE

(SANITIZED).

Data Type:
Acute (0-96 hour); Aquatic; Invertebrates

Hero ID:
4213817

Domain
Metric
Rating"!" MWF* Score Comments"*1"!"
Domain 4: Test Organism
Metric 13: Test Organism Characteristics
Metric 14: Acclimitization and Pretreatment Conditions High
Metric 15: Number of Organisms and Replicates per High
Group
Metric 16: Adequacy of Test Conditions
High	X 2	2 Daphnia magna are a recommended species in
OECD TG 202, and the source was reported as
Haskell Laboratory-bred stock.
X 1	1	Authors reported a 24 hour acclimation period.
X 1	1	Authors mentioned using 4 replicates of 5 daphnia
per concentration which is recommended by OECD
TG 202.
High	X 1	1	Test conditions were appropriate for daphnia.
Domain 5: Outcome Assessment
Metric 17: Outcome Assessment Methodology	High	x 2	2 Authors derived an EC50.
Metric 18: Consistency of Outcome Assessment	High	x 1	1 Details of outcome assessment were reported and
consistent across study groups.
Domain 6: Confounding / Variable Control
Metric 19: Confounding Variables in Test Design and High	x 2	2 No reported differences among the study group in
Procedures	environmental conditions or other factors that
could influence the outcome of the assessment.
Metric 20: Outcomes Unrelated to Exposure	High	X 1	1 No outcomes unrelated to exposure were reported.
Domain 7: Data Presentation and Analysis
Metric 21: Statistical Methods
Metric 22: Reporting of Data
Metric 23: Explanation of Unexpected Outcomes
High
High
Medium
XX X
1
2
1
1
2
2
Authors reported conducting a probit analysis.
Authors reported response rates for every test con-
centration and replicate.
Authors included a table with the number of daph-
nia exhibiting immobilization, and said that it was
cumulative. However, after 48 hours, there were
fewer immobilized daphnia than at 24 hours for the
180 mg/L concentration. Authors didn't explain this
inconsistency.
Overall Quality Determination"'"
High


1.4

Extracted
Yes




Continued on next page . . .
91

-------
. .. continued from previous page
Study Citation:
Data Type:
Hero ID:
E I Dupont Denemours & Co Inc. 1987.
(SANITIZED).
Acute (0-96 hour); Aquatic; Invertebrates
4213817
DAPHNIA MAGNA STATIC ACUTE 48-HOUR EC50 OF METHYLENE CHLORIDE
Domain
Metric
Rating"!" MWF* Score Comments"*1"!"

*	MWF = Metric Weighting Factor
t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value.
*	The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High.
Overall rating =
^ (Metric Score; x MWF,) / ^ MWF^
if any metric is Unacceptable
(round to the nearest tenth) otherwise
where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is
crossed out and an arrow points to the new rating,
ft Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments.
92

-------