PEER REVIEW DRAFT, DO NOT CITE OR QUOTE &EPA United States Environmental Protection Agency Office of Chemical Safety and Pollution Prevention Risk Evaluation for Methylene Chloride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 ------- |