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Review of 149-44-0 

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David Brooks to: Catherine, Roman 08/13/2009 08:28 AM 

Cc: Victoria Vanroden, Mark Townsend 

Hi Catherine, 

Sorry for the delay in submittal of our assessments. We have reviewed the studies submitted and our 

assessments are attached . Please note that although the robust summary attachment indicates that the 

acute health studies were not adequate, these studies were considered adequate on a weight-of-evidence 

basis due to low expected acute toxicity as well as having adequate 28-day repeated-dose, 

developmental and reproductive studies . - 

David Brooks 

Risk Assessment Division (7403M) 

High Production Volume Chemical Branch 

EPA East Room 6308N 

(202) 564-8603 

-

rr 

149-44-0 robust summary ec 6 2 09 .doc Methanesulihn acid hydroxy- monosodium salt (CAS No 149,44-0} HC Q6-19-QH.doe 
SRC TR-09-174 

SCREENING-LEVEL HAZARD CHARACTERIZATION 

OF HIGH PRODUCTION VOLUME CHEMICALS 

UNSPONSORED CHEMICAL NAME 

Methanesulfinic acid, hydroxy-, monosodium salt 

Prepared by: 

Emily Cohen, William L. Richards and Joan D. Garey 

Environmental Science Center 

Syracuse Research Corporation 

7502 Round Pond Road 

North Syracuse, NY 13212 

Contract No . EP-W-04-019 

Task FA411.4 .409.001 

Submitted to: 

OPPT/RAD/HPVCB (7403M) 

U.S . Environmental Protection Agency 

1200 Pennsylvania Avenue 

Washington, D.C. 20460 

Kathleen C. Kennedy, Project Officer 

David Brooks, Work Assignment Manager 

June 19, 2009 
SCREENING-LEVEL HAZARD CHARACTERIZATION 

OF HIGH PRODUCTION VOLUME CHEMICALS 

UNSPONSORED CHEMICAL 

Methanesulfmic acid, hydroxy-, monosodium salt (CAS No. 149-44-0) 

[9th CI Name: Methanesulfmic acid, hydroxy-, monosodium salt] 

June 2009 

Prepared by 

Risk Assessment Division 

Economics, Exposure and Technology Division 

High Production Volume Chemicals Branch 

Office of Pollution Prevention and Toxics 

Environmental Protection Agency 

1200 Pennsylvania Avenue, NW 

Washington, DC 20460-0001 
SCREENING-LEVEL HAZARD CHARACTERIZATION 

Methanesulfmic acid, hydroxy-, monosodium salt (CAS No. 149-44-0) 

Introduction 

In the HPV Challenge Program, companies have sponsored more than 2200 HPV chemicals, with approximately 

1400 chemicals sponsored directly through the HPV Challenge Program and over 860 chemicals sponsored 

indirectly through international efforts. Other chemicals, however, remained unsponsored in the voluntary 

program' . Basic hazard data for these unsponsored chemicals are being obtained through regulatory efforts such as 

TSCA Section 4 Test Rules and TSCA Section 8(a)/8(d) Rules. Methanesulfinic acid, hydroxy-, monosodium salt 

(CAS No. 149-44-0) is an unsponsored chemical and was subject to Federal Register/Vol. 71, No. 51 ; Thursday, 

March 16, 2006/page 13078. This information can be found at www.regulations .aov, under Document ID EPA-HQOPPT-2005-0033 . 

This screening-level hazard characterization is based primarily on the data available in a Data Adequacy Report on 

this chemical prepared by Oak Ridge National Laboratories (ORNQ on behalf of EPA; this report was originally 

prepared in May 2005 and updated in June 2007. In order to determine whether any new hazard information was 

developed since the time of the Data Adequacy Report, a search of the following databases was made from 2007 to 

June 2009: the NLM databases (Chem1D to locate available data sources including MedlinelPubMei, Toxline, 

HSDB, ATSDR, EPA SRS, etc.), STN/CAS online databases (Registry file for locators, ChemAbs for toxicology 

data, RTECS, Merck, etc:) and Science Direct. The structure of the unsponsored chemical is included in the 

appendix. Summary tables of SIDS endpoint data are included in the document. The screening-level hazard 

characterization for environmental and human health toxicity is based largely on SIDS endpoints and is described 

according to established EPA or OECD effect level definitions and hazard assessment practices. 

Hazard Characterization 

The evaluation of available toxicity data for fish, aquatic invertebrates and aquatic plants indicates that the potential 

hazard of methanesulfmic acid, hydroxyl-, monosodium salt to aquatic organisms is low. 

The acute oral toxicity of methanesulfinic acid, hydroxyl-, monosodium salt in rats and mice is low. In a combined 

28-day oral repeated-dose/developmental/reproductive toxicity screening test, methanesulfinic acid, hydroxyl-, 

monosodium salt produced low systemic, reproductive and developmental toxicity in rats. Methanesuifinic acid, 

hydroxyl-, monosodium salt induced chromosomal aberrations in this study. 

The gene mutation endpoint was identified as a data gap under the HPV Challenge Program. 

1. Phvsical-Chemical Properties and Environmental Fate 

The physical-chemical, environmental properties and chemical structure of methanesulfinic acid, hydroxy-, 

monosodium salt are summarized in Appendix I a. 

Physical-Chemical Properties Characterization 

Environmental Fate Characterization 

1 USEPA Regulatory Actions for Unsponsored Chemicals: http ://www epa jzov/hpv/pubs/Qeneral/reQactions .htm. 
2. Environmental Effects - Aquatic Toxicity 

Acute Toxicity to Fish 

Golden orfe (Leuciscus idus) were exposed to methanesulfinic acid, hydroxy-, monosodium salt at nominal 

concentrations of 0 (control) 5000 or 10,000 mg/L under static conditions for 96 hours. Measured concentrations 

were not provided (Nat 1987; SOCMA 2007). 

96-h LCso > 10,000 mg/L 

Acute Toxicity to Aquatic Invertebrates 

Daphnia magna were exposed to methanesulfinic acid, hydroxy-, monosodium salt at nominal concentrations of 0 (control), 10, 20, 40, 80 or 160 mg/L under static conditions for 48 hours. Measured concentrations were not 

provided. Immobilization was observed at concentrations > 20 mg/L at 48 hours (Jaeger 1999; SOCMA 2007). 

48-h ECso = 20 - 40 mg/L 

Toxicity to Aquatic Plants 

(1) Green algae (Scendesmus subspicatus) were exposed to methanesulfmic acid, hydroxyl-, monosodium salt at 

nominal concentrations of 0.1, 1, 10, 100 or 500 mg/L under static conditions for 72 hours. Measured 

concentrations were not provided (Jaeger 1999; SOCMA 2007). 

72-h EC50 (biomass) = 5 mg/L 

72-h EC50 (growth) = 450 mg/L 

(2) Green algae (Scendesmus subspicatus) were exposed to methanesulfinic acid, hydroxyl-, monosodium salt at nominal concentrations of 0, 10, 20, 40, 80, 160, 320, 640 or 1000 mg/L under static conditions for 72 hours. 

Measured concentrations were not provided (Paulus 2001 ; SOCMA 2007;) . 72-h EC50 (biomass) = 220 mg/L 

72-h EC50 (growth) = 370mg/L 

Conclusion: The evaluation of available toxicity data for fish, aquatic invertebrates and aquatic plants indicates that the potential hazard of methanesulfinic acid, hydroxyl-, monosodium salt to aquatic organisms is low. 

3. Human Health Effects 

Acute Oral Toxicity 

(1) Rats (stain and sex/dose not specified) were administered methanesulfinic acid, hydroxyl-, monosodium salt via 

an unspecified oral route at 2000 mg/kg. No other information was provided (Author unknown 1980). 

LDso > 2000 mg/kg-bw 

(2) Mice (strain and sex/dose not specified) were administered methanesulfinic acid, hydroxyl-, monosodium salt 

via an unspecified oral route at 4000 mg/kg-bw. No other information was provided (Author unknown 1980). 

LDso = 4000 mg/kg-bw 

Repeated Dose/Reproductive/Developmental Toxicity 

In a combined repeated-dose/reproductive/developmental toxicity screening test, Wistar rats (10/sex/group) were 

administered methanesulfinic acid, hydroxyl-, monosodium salt via oral gavage at 0, 100, 300 or 1000 mg/kg/day in 

water. Males were exposed for 30 days (during 2 weeks prior to mating, during mating, and to termination). 

Females were exposed for 42 - 53 days (during 2 weeks prior to mating, during mating and gestation, up to day 4 of 

lactation) . Parental effects at 1000 mg/kg-bw/day included hunched posture, piloerection, slight salivation, 

decreased body weight, decreased food consumption, hematological effects (increased white blood cell counts in 

males and females and increased relative numbers of reticulocytes in males) and changes in clinical chemistry 

(increased alanine aminotransferase in males and females and increased activity of aspartate aminotransferse in one 
male). Increases in organ/body weight ratios for the liver, spleen, testes and epididymides of males and liver, 

spleen, kidneys and adrenals of females were observed. There were no corresponding histological changes. 

Increased organ/body weight ratio of the brain was observed in males and absolute brain weight was slightly 

decreased in females. No treatment related effects were observed at the mid- or low-dose levels. No treatmentrelated reproductive or developmental effects were observed (Beekhuijzen 2008). 

LOAEL (systemic toxicity) = 1000 mg/kg-bw/day (based on effects on clinical signs of toxicity, body weight, 

food consumption, hematology, clinical chemistry and organ weight) 

NOAEL (systemic toxicity) = 300 mg/kg-bw/day 

NOAEL (reproductive and developmental toxicity) = 1000 mg/kg-bw/day (based on no effects at the highest 

dose tested) 

Genetic Toxicity - Gene Mutation 

No adequate data were provided. Testing is recommended. 

Genetic Toxicity - Chromosomal Aberrations 

In vivo 

In a micronucleus test in bone marrow cells, NMRI mice (5/sex/dose) were administered methanesulfinic acid, 

hydroxyl-, monosodium salt via the intraperitoneal route in physiological saline at 2000 (two groups), 1000 (one 

group), and 500 (one group) mg/kg-bw. Bone marrow was sampled 24 or 48 hours (highest dose only) after dosing . Negative and positive control substances produced appropriate responses. Methanesulfinic acid, hydroxy-, 

monosodium salt induced an increase in the mean number of micronucleated polychromatic erythrocytes but showed 

no decrease in the ratio of polychromatic to normochromatic erythrocytes (Buskens 2008). 

Methanesulfinic acid, hydroxyl-, monosodium salt induced chromosomal aberrations in this study. 

Conclusion: The acute oral toxicity of methanesulfinic acid, hydroxyl-, monosodium salt in rats and mice is low. In a combined 28-day oral repeated-dose/developmental/reproductive toxicity screening test, methanesulfinic acid, 

hydroxyl-, monosodium salt produced low systemic, reproductive and developmental toxicity in rats . 

Methanesulfinic acid, hydroxyl-, monosodium salt induced chromosomal aberrations in this assay. The gene 

mutation endpoint remains as a data gap. 

4. Hazard Characterization 

5. Data Gaps 
APPENDIX 1 

Summary Table of the Screening Information Data Set 

as Submitted under the U.S . HPV Challenge Program 

Endpoints SPONSORED CHEMICAL 

Methanesulf'mic acid, hydroxy-, monosodium salt 

(149-44-0) 

Structure 

Summary of Physical-Chemical Properties and Environmental Fate Data 

Melting Point (°C) 

Boiling Point (°C) 

Vapor Pressure 

(hPa at 25°C) 

Log Kow 

Water Solubility 

(mg/L at 25°C) 

Direct Photodegradation 

Indirect (OH-) Photodegradation 

Half-life (t1,2) 

Stability in Water (Hydrolysis) (ti/2) 

Fugacity 

(Level HI Model) 

Biodegradation at 28 days (%) 

Air (%) 

Water (%) 

Soil (%) 

Sediment (%) 

Summary of Environmental Effects - Aquatic Toxicity Data 

Fish 

96-h LCso (mg/L) > 10,000 

Aquatic Invertebrates 20-40 48-h EC5o (mg/L) 

Aquatic Plants 

72-h ECso (mg/L) 

(growth) 370 

(biomass) 
Summary Table of the Screening Information Data Set 

as Submitted under the U.S. HPV Challenge Program 

Endpoints SPONSORED CHEMICAL 

Methanesulfmic acid, hydroxy-, monosodium salt 

(149-44-0) 

Summary of Human Health Data 

Acute Oral Toxicity 

LDso (mg/kg-bw) > 2000 

Repeated-Dose/Reproductive/Developmental Toxicity 

NOAEL/LOAEL 

Oral (mg/kg-bw/day) 

Systemic Toxicity NOAEL = 300 

LOAEL = 1000 

Reproductive and Developmental Toxicity NOAEL = 1000 (highest dose tested) 

~Genetic Toxicity - Gene Mutation No adequate data. Testing is recommended. 

In vitro 

Genetic Toxicity - Chromosomal Aberrations Positive 

En vivo 

6. Additional Comments 

Acute oral toxicity studies in rats and mice were provided. These studies were considered adequate on a weight-ofevidence basis due to low expected toxicity (LDso > 2000 mg/kg-bw). 

An in vivo sex linked recessive lethal test in Drosophila was provided by the sponsor. This study was not included in the hazard characterization because it received a Klimisch code of 3 . 

One TSCATS document was found in the 8(e) initial screen database (8EHQ-0508_17139A, Document No. : 880080000182). This document contained results of a micronucleus test in mice. The primary document was not 

included with the submission, but was included in a separate submission by NOTOX. This study was included in 

the hazard characterization document. 

7. References 

Author unknown. 1980. Title not given. Yakkyoku Pharmcy. 31 :959, as cited in RTECS. REFERENCE 

ORDERED. This study was considered acceptable on a weight of evidence basis. 

Beekhuijzen, M.E .W. 2008 . A combined 28-day repeated dose toxicity study with the reproduction/developmental 

toxicity screening test of methane sulfmic acid, hydroxy-, monosodium salt in rats by oral gavage. NOTOX B.V. 

Report No. 485218 (485615), dated 01-15-2008 (first draft of an unpublished study report); Robust Summaries and Dossier for CAS No. 149-44-0, PCA Services, Inc. 

Buskens, C.A.F . 2008. Study Title: Micronucleus test in bone marrow cells of the mouse with methane sulfinic acid, hydroxy-, monosodium salt C.A.F . Buskens, Study completion date : 22 May 2008. NOTOX B.V., NOTOX Project 485472; The robust summary for this study was also provided in: Robust Summaries and Dossier for CAS No. 149-44-0, PCA Services, Inc. 

Jaeger, W. 1999 . Acute toxicity of Daphnia according to OECD 202 Part 1 for sodium hydroxymethanesulfinate. Analytik GmbH study dates 1999-05-12 (English translation of an unpublished German language study); Robust Summaries and Dossier for CAS No. 149-44-0, PCA Services, Inc. 
Nat, R. 1987. Report on the study of the acute toxicity: Ronalgit CBlankit C, BASF Aktiengesellschaft 

Department of Toxicology, Report No. 10F033/875145, dated 1987-09-17; Robust Summaries and Dossier for CAS 

No. 149-44-0, PCA Services, Inc. 

Paulus, J. 2001 . Determination of the 72 hour EC50 of "Brueggolit E02" in the presence of Scenedesmus 

subspicatus. LAUS-Libertaere Arbeitsgemeinschaft Environmental & Hazard Substance Consultation GmbH 

Report No. 00110605J01, dated 2001-02-21 . (English translation of an unpublished German language study) ; 

Robust Summaries and Dossier for CAS No. 149-44-0, PCA Services, Inc. 

SOCMA. 2007. Synthetic Organic Chemical Manufacturers Association. Submission dated May 14, 2007. 

Submission is attached and can also be found at regulations.gov, Docket ID is EPA-HQ-OPPT-2005-0033-0238 . 
APPENDIX 2 

CHECKLIST FOR OTHER SOURCES OF DATA ON HPV CHEMICALS 

CHEMICAL NAME: Methanesulfinic acid, hydroxy-, monosodium salt CAS #: .149-44-0 

Checker (Name) : Emily Cohen Date: March 25, 2009 

CAS# CAS# 

NOT Found 

Found 

® USEPA - TSCATS http ://www.syrres.com/esc/tscats htm 

NOTES: One TSCATS document was found in the 8(e) initial screen 

database: 8ehcL0508-17139A, DCN #: 880080000182 

USEPA - OPP (Office of Pesticide Programs) - working on a unified 

searchable list 

1) http://www.ena.gov/oppsrrdl/registration review/schedule htm 

2) http://www.epa.gov/pesticides/reregistration/status htm 

NOTES: 

® USEPA - Inert Chemicals 

Complete List by CAS: http://www.epa. ov/opprd001/inerts/lists html 

Definition of List # http ://www.e a. ov/opprd001/inerts/lists html 

Reassessment Status : http://www epa ov/o prd001/inerts/decisiondoc_a2k html 

0 List 1- EPA's Office of Pesticide Programs determined Inert Ingredients of 

Toxicological Concern. 

0 List 2 - Potentially Toxic Other Ingredients/High Priority for Testing inerts . 

® List 3 - Inerts of unknown toxicity. 

0 List 4A - Minimal risk inert ingredients. 

0 List 4B - Other ingredients for which EPA has sufficient information to 

reasonably conclude that the current use pattern in pesticide products will not 

adversely affect public health or the environment. 

NOTES: List 3 - Inerts of unknown toxicity. 

0 USEPA - IRIS (Integrated Risk Information System) 

http://www.ena .gov/ncea/iris/search keyword htm 

NOTES: 

0 USEPA - TRI (Toxics Release Inventory Chemicals) 

http://www.epa.gov/enviro/html/tris/tris query html - Chemical Name Search has 

CAS# Option 
http ://www.atsdr.cdc_gov/toxfaq.html - ATSDR Tox FAQs, which are very brief 

summaries of the Tox Profiles 

NOTES: 

USEPA - Ambient Water Quality Criteria (Both Human Health and 

Aquatic Life) 

http ://www epa gov/waterscience/criteria/wqcriteria .html 

There is no search engine ; There are three different lists (Priority Pollutant, NonPriority Pollutant, and Orizanoleptic ; scan each list carefully - they are not in strict 

alphabetical order. 

NOTES: 

0 USEPA - Drinking Water Standards; Alphabetical List, No CAS#s 

http //www epa g.ov/safewater/contaminants/index .html 

NOTES: 

® USEPA - ECOTOX http ://www.epa.gov\ecotox 

NOTES: One aquatic study found: MacPhee, C., and R. Ruelle, Lethal Effects of 

1888 Chemicals upon Four Species of Fish from Western North America, 

Bu1l.No.3, Forest, Wildl. and Range Exp.Stn ., Univ.of Idaho, Moscow, ID :112 

p., 1969. Reference number: 15148 

Terrestrial report : 1 study found: Asada, K., and Z. Kasai, Inhibition of the 

Photosynthetic Carbon Dioxide Fixation of Green Plants by AHydroxysulfonates, and Its Effects on the Assimilation Products, Plant Cel P 

3 :125-136, 1962. Reference number: 40903 

0 NTP (National Toxicology Program) 

ht ://n - 

apps niehs nihgo-v_/ntp tox/index cfin?fuseaction=ntpsearch .searchhome 

NOTES: 

0 ATSDR (Agency for Toxic Substances & Disease Registry) 

httL'//www atsdr cdc o

~ 

v/toxpro2 .htm1#bookmark05 

NOTES: 

0 IPCS - CICADs (Concise International Chemical Assessment Documents) 

http'//www incbem.org/pages/cicads.html 

NOTES: 
1 SRC TR-09-033 

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9 ORPHAN CHEMICAL DATA ADEQUACY REVIEW AND ROBUST SUMMARY 

10 ADEQUACY REQUIREMENTS 

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12 Methanesulfinic acid, hydroxyl-, monosodium salt 

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14 Prepared by: 

15 

16 Emily Cohen 

17 

18 Environmental Science Center 

19 Syracuse Research Corporation 

20 7502 Round Pond Road 

21 North Syracuse, NY 13212 

22 

23 

24 Contract No. EP-W-04-019 

25 Task FA411 .4.409.004 

26 

27 

28 Submitted to : 

29 

30 OPPT/RAD/HPVCB (7403M) 

31 U.S . Environmental Protection Agency 

32 1200 Pennsylvania Avenue 

33 Washington, D.C. 20460 

34 

35 Kathleen C. Kennedy, Project Officer 

36 David Brooks, Work Assignment Manager 

37 

38 June 2, 2009 

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41 Orphan Chemical Data Adequacy Review and Robust Summary Adequacy Requirements 

42 Chemical Name: 

43 Methanesulfinic acid, hydroxyl-, monosodium salt 

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60 Submitted to: 

61 High Production Volume Chemicals Branch 

62 Risk Assessment Division 

63 Office of Pollution Prevention and Toxics 

64 Environmental Protection Agency 

65 WAM: David Brooks 

' 66 1200 Pennsylvania Avenue, NW 

67 Washington, DC 20460-0001 

68 

69 Prepared By: 

70 SRC 

71 Environmental Science Center 

72 2451 Crystal Drive, Suite 804 

73 Arlington, VA 22202 

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ACUTE TOXICITY TO FISH 

Summary of endpoint: Adequate 

Title: Acute Toxicity to Aquatic Vertebrates 

The study is : ® Adequate 

TEST SUBSTANCE 

0 Not Adequate 

Identity: Methanesulfinic acid, hydroxyl-, monosodium salt (CAS No . 149-44-0) 

Remarks: Not stated 

METHOD 

Method/guideline followed: DIN 38 412 

Test Type: Not stated 

GLP (Y/N) : Not stated 

Analytical monitoring: Not stated 

Year study performed: 2007 

Species: Golden Orfe, Leuciscus idus 

Exposure period: 48 hours 

Limit test : Not stated 

Dose/Concentration tested : 5000 and 10,000 mg/L 

Method remarks: 

Total Hardness: 2.5 MMOL/L 

Acid Capacity: 0.8 MMOL/L 

Ratio CA/MG Ions: 4:1 

Ratio NA/If Ions: 10:1 

Volume of Water: 10 liters 

No. of animals per test concentration: 10 

Loading (G Fish/L Test Water): 2.4 

Test Vessels : All-glass Aquarium (30 cm x 22 cm x 24 cm) 

Adaptation to test water and test temperature: 3 days 

Withdrawal of food: 1 day 
122 

123 Water chemistry in the test: 

124 pH value: - 7.7 

125 Test Temperature and Units: 20 degrees Celsius 

126 Photo (light/Dark) : 16/8 

127 

128 RESULTS 

129 

130 Detailed results : 

131 Maximum concentration causing no mortality: 10000 MG/L 

132 Minimum concentration causing 100% mortality : > 10000 MG/L 

133 

134 Nominal Conc./Number of fish/Dead Fish After 

135 (MG/L)/1 H / 4 H / 24H / 48H / 72H / 96H 

136 5000/ 10/0/0/0/0/0/0 

137 10000/10/0/0/0/0/0/0 

138 0/10/0/0/0/0/0/0 

139 

140 No symptoms detectable 

141 

142 The chemical was not acutely toxic to Leuciscus idus at concentrations up to 10,000 mg/L 

143 (Soma, 2007) 

144 

145 * See report : "SOCMA submission-May 14, 2007, pp.19-26 ." 

146 

147 CONCLUSIONS 

148 

149 NOEC = 10000 mg/L 

150 

151 RELIABILITY CODE 

152 

153 1- Valid Without Restrictions 

154 German national test guide study. 

155 Critical study. 

156 Key 

157 

158 REFERENCES 

159 

160 SOCMA. 2007. Synthetic Organic Chemical Manufacturers Association. Submission dated May 

161 14, 2007. Submission is attached and can also be found at regulations. gov, Docket ID is EPA162 HQ-OPPT-2005-0033-0238 . 

163 

164 

165 

166 

167 
168 Title: Acute Toxicity to Aquatic Vertebrates 

169 

170 The study is : ® Adequate F-1 Not Adequate 

171 

172 TEST SUBSTANCE 

173 

174 Identity: Methanesulfinic acid, hydroxyl-, monosodium salt (CAS No. 149-44-0) 

175 

176 Remarks: The test material was a commercial formulation of methanesulfinic acid, hydroxyl-, 

177 monosodium salt (Rongalit CBlankit C). The purity of the formulation was not listed . 

178 

179 METHOD 

180 

181 Method/guideline followed: Other: DIN38 412 

182 

183 Test Type: Static 

184 

185 GLP (Y/N): No data 

186 

187 Analytical monitoring: No 

188 

189 Year study performed: 1987 

190 

191 Species : Leuciscus idus 

192 

193 Exposure period: 96 hours 

194 

195 Limit test: Not stated 

196 

197 Dose/Concentration tested: 5,000 or 10,000 mg/1 

198 

199 Method remarks: 

200 Water media type: freshwater 

201 Test concentration : Nominal 

202 

203 Test water was reconstituted freshwater, prepared from fully demineralized tap water according 

204 to the guideline. The maximum conductivity was 10 micro Mho After resalting, the hardness, 

205 acid capacity, ratio of Ca/Mg ions, ratio of Na/K ions and pH were 2.5 mmol/1, 0.8 mmol/1, 4:1, 

206 10 :1 and approximately 7.7 . 

207 

208 Fish were acclimated to test water for 3 days prior to exposure. The photoperiod was 16 hours 

209 light and 8 hours darkness. Food (type not specified) was withdrawn 1 day prior to testing . No 

210 other details were provided. 

211 

212 Test material (5,000 or 10,000 mg/1) was added to the test water (10 liters/vessel) without 

213 pretreatment . The temperature, pH and hardness of the water were 20 degrees C, approximately 
214 7.7, and 2.5 mmol/1, respectively . Fish (10/concentration, 2.4 g fish/liter test water) were added 

215 to the aquaria (30 cm x 22 cm x 24 cm all-glass aquaria). The medium was continuously aerated 

216 with oil-free air throughout the test. 

217 

218 RESULTS 

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220 Detailed results : 

221 None of the animals died and no symptoms of toxicity were reported. 

222 

223 CONCLUSIONS 

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225 96 hour LC50 > 10,000 mg/L 

226 

227 RELIABILITY CODE 

228 

229 1- Valid Without Restrictions 

230 Critical study for SIDS endpoint 

231 

232 REFERENCES 

233 

234 Nat, R. 1987. Report on the Study of the Acute Toxicity: Ronalgit CBlankit C, BASF 

235 Aktiengesellschaft Department of Toxicology, Report No . l OF033/875145, dated 1987-09-17 

236 

237 Robust Summaries and Dossier for CAS No. 149-44-0, PCA Services, Inc. 

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-260 ACUTE TOXICITY TO DAPHNIA 

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262 Summary of endpoint: Adequate 

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264 Title: Acute Toxicity to Aquatic Invertebrates 

265 

266 The study is: ® Adequate ~ Not Adequate 

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268 TEST SUBSTANCE 

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270 Identity: Methanesulfinic acid, hydroxyl-, monosodium salt (CAS No. 149-44-0) 

271 

272 Remarks: Sodiumhydroxymethanesulfinate, CAS No. : 149-44-0, Preparation, solid white 

273 powder 

274 

275 Note: CAS No. given in report is incorrectly listed as 497-19-8 which is the CAS No. for sodium 

276 carbonate. 

277 

278 METHOD 

279 

280 Method/guideline followed: OECD 202 

281 

282 Test Type: Static 

283 

284 GLP (Y/N): Not stated 

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286 Analytical monitoring: Not stated 

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288 Year study performed: 2007 

289 

290 Species : Daphnia magna 

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292 Exposure period: 48 hours 

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294 Limit test: Not stated 

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296 Dose/Concentration tested: Not stated 

297 

298 Method remarks: 

299 Water solubility: ca. 600 g/L 

300 Solvent : water 

301 

302 The objective of the study is to determine the acute toxicity of sodium hydroxymethanesulfinate 

303 with respect to daphnia. 

304 

305 The acute toxicity is understood for this test method to be the median effective concentration 
306 (EC50) for the influence on reduction of swimming ability, where 50% of the daphnia in a test 

307 group are rendered unable to swim within a certain time period of continuous exposure to the test 

308 substance. 

309 

310 Test Method: Acute toxicity for Daphnia, OECD 202, Part 1 

311 SOP-CP-009, acute toxicity for daphnia, (Daphnia magna) 

312 

313 Duration: 48 hours 

314 Type test: Static (no water exchange) 

315 

316 Test organism 

317 Type: Daphnia magna, Clone 5 

318 Crustacea, Phyllopoda, Cladocera, Daphnidae 

319 Source: Office of the Environment of the Bundles Republic Germany 

320 

321 Storage and Handling 

322 The food was a pure culture of the green algae Scendesmus suspicatus (3x daily via a tube 

323 pump). The animals were transferred 3x per week to a new medium via a sieve. Approx. every 4-

324 6 weeks the old animals were replaced with young animals on the basis of ongoing reproduction. 

325 

326 Nutrient Medium 

327 The nourishment medium was made up as follows with deionized water 

328 

329 Synthetic seawater salt (Wimex prepared seawater salt) 0.333 g/L 

330 CaC12 x 2 H20 (C: 117.6 g/L) 2 .3 mL/L 

331 NaHC03 (C : 25 .2 g/L) 2.2 mL/L 

332 Se02 (C: 1 .4 g/L) 0.1 mL/L 

333 

334 The Medium was aerated ahead of time to the saturation point of oxygen and adjusted to a pH 

335 value of 7.8 ± 0.2 . The synthetic seawater salt contained 7 macronutrient materials, 60 trace 

336 elements, 8 vitamins and 3 amino acids. Through the addition of CaC12 and NaHC03 the total 

337 hardness (2.5 mmol/L), the Ca/Mg-ratio (4:1), and the alkalinity (0.8 mmol/L) was adjusted 

338 according to the medium recommended by the OECD Guideline 202 . 

339 

340 Test Conditions 

341 

342 Animals: 20 animals per concentration (4 x 5 in each 20 mL, 50 mL glass beakers) 

343 Aeration: none 

344 

345 Oxygen Content 

346 

347 Table 1 Dissolved Oxygen Content (%) 

348 

349 Concentration (mg/L) 

350 Time (hr)/Control/10/20/40/80/160 

351 0/98/98/98/98/98/98 
352 48/96/96/96/96/97/97 

353 

354 Table 2: pH Value 

355 

356 Concentration (mg/L) 

357 Time (hr)/Control/10/20/40/80/160 

358 0/7.75/7.75/7.76/7.77/7.82/7.92 

359 48/7 .81/7.85/7.86/7.86/7.87/7.88 

360 

361 Table 3 : Immobilization and Number of Animals Hanging on Water Surface 

362 

363 Concentration (mg/L) 

364 Control/ 10/20/40/80/160 

365 24 hour immobilization (%)/0/0/5/0/0/5 

366 48 hour immobilization (%)/0/0/15/65/75/90 

367 Animals on Water/Surface (%)/0/0/0/0/0/0 

368 

369 RESULTS 

370 

371 Detailed results : 

372 Conclusion: The test substance "Sodiumhydroxymethanesulfinate" shows the following effect on 

373 the mobility of the animals in a test room after 48 hours: 

374 

375 EC50 (48 hours: 50 mg/L) 

376 

377 A 48 hour LC50 of 50 mg/L was reported by SOMA (2007), but the data presented indicate that 

378 the LC50 was between 20 to 40 mg/L. 

379 

380 * See report : SOCMA submission-May 14, 2007, pp. 27-34. 

381 

382 CONCLUSIONS 

383 

384 24 hour EC50 > 150 mg/L 

385 48 hour EC50 = 50 mg/L 

386 

387 RELIABILITY CODE 

388 

389 1- Valid Without Restrictions 

390 Guideline study (OECD TG 202 Part 1) . 

391 Critical study, Key 

392 

393 REFERENCES 

394 

395 SOCMA. 2007. Synthetic Organic Chemical Manufacturers Association. Submission dated May 

396 14, 2007. Submission is attached and can also be found at regulations.gov. Docket ID is EPA397 HQ-OPPT-2005-0033-0238 . 
398 Title: Acute Toxicity to Aquatic Invertebrates 

399 

400 The study is: ® Adequate 0 Not Adequate 

401 

402 TEST SUBSTANCE 

403 

404 Identity: Methanesulfinic acid, hydroxyl-, monosodium salt (CAS No. 149-44-0) 

405 

406 Remarks: as prescribed by 1 .1-1 .4, The test material was incorrectly identified as CAS No. 497-

407 19-8 in the study report . The chemical is a commercial product containing sodium 

408 hydroxymethanesulfinate and 1-3% CAS No. 497-19-8 (sodium carbonate). The exact purity is 

409 unknown. 

410 

411 METHOD 

412 

413 Method/guideline followed: OECD 202 

414 

415 Test Type: Static 

416 

417 GLP (Y/N) : Yes 

418 

419 Analytical monitoring: No 

420 

421 Year study performed: 1999 

422 

423 Species : Daphnia magna 

424 

425 Exposure period: 48 hours 

426 

427 Limit test: Not stated 

428 

429 Dose/Concentration tested : Concentrations tested were 10 mg/L, 20 mg/L, 40 mg/L, 80 mg/L, 

430 and 160 mg/L. 

431 

432 Method remarks : 

433 Water media type: Freshwater 

434 Test Concentrations : Nominal 

435 

436 The data came from an English translation of a German language study. 

437 

438 The test was valid as = 10% of the controls were immobilized or floating on the water surface, 

439 the concentration of the dissolved oxygen in the solutions was > 60% of the air saturation 

44o program for the entire time of exposure and the pH values did not fall more than one unit. 

441 

442 The storage of the animals was at a constant temperature of 23 +/- 1 C. The photoperiod was 14 

443 hours. The food ws a pure culture of the green algae Scenedesmus suspicatus (3x daily via a 
444 tube pump). The animals were transferred 3x per week to a new medium via a sieve. Approx. 

445 every 4-6 weeks the old animals were replaced with young animals on the basis of ongoing 

446 reproduction. 

447 

448 The medium was aerated ahead of time to the saturation point of oxygen and adjusted to a pH 

449 value of 7.8 +/- 0.2 . The symthetic seawater salt contained 7 macronutrient materials, 60 trace 

450 elements, 8 vitamins and 3 amino acids. Through the addition of CaCl2 and NaHC03 the total 

451 hardness (2.5 mmol/L), the Ca/Mg-ratio (4:1), and the alkalinity (0.8 mmol/L) was adjusted 

452 according to the medium recommended by the OECD Guideline 202. 

453 

454 Twenty animals were tested per concentration (4x5 in each 20 mL, 50 mL glass beakers), at 20 

455 +/- 1 C. Concentrations tested were 10 mg/L, 20 mg/L, 40 mg/L, 80 mg/L, and 160 mg/L. The 

456 photoperiod was 14 hours. The pH value, oxygen content and temperature were determined at 

457 the beginning and at the end of the 48-hr test period. The medium was not aerated. The 

458 measurements at Time 0 were conducted in an extra run without animals in order to avoid a 

459 negative influence through the exchange of electrodes. 

460 

461 The following parameters were observed at 0, 24 and 48 hours: immobilized animals, 

462 immobilized animals that float on the water's surface, animals not immobilized, but that hang on 

463 the surface of the water, and animals not immobilized, but that are weak and gravitate to the 

464 bottom of the test container . 

465 

466 RESULTS 

467 

468 Detailed results: 

469 Dissolved Oxygen Content ranged from 96%-98% for each concentration at 0 and 48 hours. pH 

470 values ranged from 7.75 to 7.92 . 

471 

472 Immobilization and Number of Animals Hanging on Water Surface 

473 Concentration (mg/L) 

474 Time/Control/10/20/40/80/160 

475 24 hour (%)/0/0/5/0/0/5 

476 48 hour (%)/0/0/15/65/75/90 

477 Animals on water surface (%)/0/0/0/0/0/0 

478 

479 CONCLUSIONS 

480 

481 48 hour EC50 = 20-40 mg/L 

482 

483 RELIABILITY CODE 

484 

485 1- Valid Without Restrictions 

486 Flag: Critical study for SIDS endpoint 

487 

488 

489 
49o REFERENCES 

491 

492 Jaeger, W. 1999. Acute Toxicity of Daphnia According to OECD 202 Part 1 for Sodium 

493 hydroxymethanesulfinate. Analytik GmbH study dates 1999-05-12 (English translation of an 

494 unpublished German language study 

495 

496 Robust Summaries and Dossier for CAS No. 149-44-0, PCA Services, Inc. 

497 

498 

499 

500 

501 

502 

503 

504 

505 

506 

507 

508 

509 

510 

511 

512 

513 

514 

515 

516 

517 

518 

519 

520 

521 

522 

523 

524 

525 

526 

527 

528 

529 

530 

531 

532 

533 

534 

535 
536 

537 

538 TOXICITY TO AQUATIC PLANTS 

539 

540 Summary of endpoint: Adequate 

541 

542 Title: Toxicity to Aquatic Plants e.g. Algae 

543 

544 The study is : ® Adequate ~ Not Adequate 

545 

546 TEST SUBSTANCE 

547 

548 Identity: Methanesulfinic acid, hydroxyl-, monosodium salt (CAS No. 149-44-0) 

549 

550 Remarks: Sodiumhydroxymethanesulfinate 

551 

552 METHOD 

553 

554 Method/guideline followed: OECD 201 . 

555 

556 Test Type (endpoint) : Not stated 

557 

558 GLP (Y/N): Yes 

559 

560 Analytical monitoring: Not stated 

561 

562 Year study performed: 2007 

563 

564 Species: Scenedesmus subspicatus (algae) 

565 

566 Exposure period: 72 hours 

567 

568 Limit test: Not stated 

569 

570 Dose/Concentration tested: 8 concentrations (10000-10 mg/1) were tested 

571 

572 Remark: This study has been carried out with the following protocol guidelines 

573 

574 - OECD Guidelines for the Testing of Chemicals "Alga, Growth Inhibition Test" succeeded by 

575 OECD Nr. 201 on 07 .June 1984. 

576 - EN28692 "Determination of acute Toxicity of Water, Wastewater and Water soluble chemicals 

577 versus Scenedesmus subspicatus Chodat." 

578 - SOP 118 003 01 Revised Edition 3 of 01 .10.1999 of LAUS GmbH "Determination of Toxicity 

579 toward Scenedesmus subspicatus Chodat." 

580 

581 Every 3 days before the start of the preliminary and main tests a batch of algae was taken out of 
582 the stem culture (Ch.B. 210600), the algae were apportioned in a culture flask with 50 ml 

583 foreculture medium (Ch.B. 11122000). This was incubated for 72 hours with stirring and 

584 lighting. After adjusting the cell number at about l 0E5/ml by means of a photometric 

585 measurement and the addition of fore culture medium the culture was ready for the test. 

586 

587 Preliminary preparations 

588 

589 On the day before the start of the test a sufficient number of test vessels was readied. Two hours 

590 before the test start the MicroGreeny® was prewarmed. Next the stem solution of the dilution 

591 series (concentration in the realm of 1000 to 0.1 mg/1) was prepared and pipetted into the test 

592 vessel (4.00 ml each), such that a mixture of the added foreculture and nutrient concentrate was 

593 achieved (every 0.50 ml, produced 1 .00 ml) 

594 

595 Test 

596 

597 Date of the test run 08-11 .02.2001 

598 Test Vessel : Screwtop tubelets d-6 mm 

599 Test duration : 72 hours 

600 Test Temperature: 21±2°C 

601 Control : Demineralized water 

602 Control : Demineralized water 

603 

604 The test vessels were incubated open under stirring (magnet stirrers) at 80001ux lighting. Every 

605 24 hours the cell number was determined by means of photometric measurement at 440 nn) . 

606 Before and after the test the pH values of the control were measured. The measurement values of 

607 the optical thickness were recorded in the formular, and after the conclusion of the test were 

608 evaluated with the program Origin (TM) . 

609 

610 Main Test 

611 Preparation Analog preliminary test/8 concentrations (10000-10 mg/1) were tested 

612 

613 Test 

614 Date of test conduct: 13-16.02 .2001 

615 Test Vessels: Screwtop tubeletts d-6mm 

616 Test duration : 72 hours 

617 Test Temperature : 21±2°C 

618 Control : demineralized water 

619 

620 The conduct of the test was along the same lines of the preliminary test. 

621 

622 RESULTS 

623 

624 Detailed results: 

625 Preliminary test 

626 Tabular Presentation of the Extinctions 

627 
628 Concentration in (mg/1)/E(Oh)/E(24h)/E(48h)/E(72h) 

629 0/14/66/186/330 

630 0 /14/68/190/333 

631 0/12/66/192/333 

632 1000/16/24/18/21 

633 1000/16/18/17/21 

634 1000/16/21/19/20 

635 100/14/63/135/228 

636 100/16/63/134/220 

637 100/12/55/128/209 

638 10 /14/66/191/342 

639 10 /17/66/186/334 

640 10 /16/68/185/331 

641 1/16/79/207/248 

642 1 /19/82/208/347 

643 1/16/72/192/341 

644 0.1/13/65/195/337 

645 0.1/15 /65/191/328 

646 0.1 /13 /63/187/326 

647 

648 The measured extinctions and the inherent coloration of the mixture were corrected for the 

649 calculation of the biomass integrals and the growth rate . 

650 

651 Main Test 

652 

653 Tabular presentation of Extinctions 

654 

655 Concentration in (mg/1)/E(Oh)/E(24h)/E(48h)/E(72h) 

656 0/14/49/142/299 

657 0/13/48/139/289 

658 0/12/44/148/292 

659 10/14/45/145/289 

660 10 /12/46/147/264 

661 10 /13/45/151/279 

662 20/14/47/141/271 

663 20 /14/44/137/274 

664 20/13/45/138/284 

665 40/12/43/141/267 

666 40 /14/47/145/278 

667 40 /13/46/140/273 

668 80/14/42/138/264 

669 80 /13/42/137/251 

670 80/14/44/138/260 

671 160 /14/39/101/133 

672 160/12/36/108/133 

673 160 /11/37/98/120 
674 320 /13/29/58/85 

675 320 /15 /23/59/86 

676 320 /13/31/67/83 

677 640 /13/19/25/31 

678 640/12/22/28/33 

679 640 /14/22/26/32 

680 1000/13/22/23/26 

681 1000 /13/20/21/25 

682 1000 /13/70/75/87 

683 

684 The measured extinctions and the inherent coloration of the mixture were corrected for the 

685 calculation of the biomass integrals and the growth rate . 

686 

687 Results obtained 

688 

689 Parameter Value 95% Confidence Range 

690 NOEL 72 h 10 mg/1 Not determined 

691 72 h ErC50 370 mg/1 170-680 mg/1 

692 72 h EbC50 220 mg/1 90-450 mg/1 

693 72 h Erc 100 > 1000 mg/1 Not determined 

694 72 h EbC 100 > 1000 mg/1 Not determined 

695 

696 Growth rate 

697 

698 The nominal growth factor of 16 hours within 72 hours that is expected for the control run 

699 corresponds to a growth rate of 0.9 . The control for the preliminary test had a growth rate of 

700 1 .07, and the one for the main test had a growth rate of 1 .04. 

701 

702 pH Variation 

703 

704 The pH value of the control run did not change more than 1 .5 units. In the preliminary test the 

705 variation was 0.5 ; likewise in the main test it was 0.5 units. 

706 

707 Two studies using Scenedesmus subspicatus were submitted by SOCMA (2007). The first study 

708 identified an ErC50 of 370 mg/L and an EbC50 of 220 mg/L. The second study reported an 

709 ErC50 of 450 mg/L and an EbC50 of 5 mg/L. 

710 

711 * See report: SOCMA submission-May 14, 2007, pp. 43-60, 79-90. 

712 

713 CONCLUSIONS 

714 

715 Parameter Value 95% Confidence Range 

716 NOEL 72 h 10 mg/1 Not determined 

717 72 h ErC50 370 mg/1 170-680 mg/1 

718 72 h EbC50 220 mg/1 90-450 mg/1 

719 72 h Erc 100 > 1000 mg/1 Not determined 
'72o 72 h EbC l 00 > 1000 mg/1 Not determined 

721 

722 RELIABILITY CODE 

723 

724 1- Valid Without Restrictions 

725 Guideline Study (OECD 201). 

726 Critical study. Key 

727 

728 REFERENCES 

729 

730 SOCMA. 2007. Synthetic Organic Chemical Manufacturers Association. Submission dated May 

731 14, 2007. Submission is attached and can also be found at regulations. gov. Docket ID is EPA732 HQ-OPPT-2005-0033-0238 . 

733 

734 

735 

736 

737 

738 

739 

740 

741 

742 

743 

744 

745 

746 

747 

748 

749 

750 

751 

752 

753 

754 

755 

756 

757 

758 

759 

760 

761 

762 

763 

764 

765 
766 

767 

768 Title: Toxicity to Aquatic Plants e.g . Algae 

769 

770 The study is: ® Adequate ~ Not Adequate 

771 

772 TEST SUBSTANCE 

773 

774 Identity: Methanesulfinic acid, hydroxyl-, monosodium salt (CAS No. 149-44-0) 

775 

776 Remarks: as prescribed by 1 .1-1 .4, Brueggolite E02 (a commercial sodium 

777 hydroxymethanesulfinate), Batch 02112000 from Brueggemann. The purity of the formulation 

778 was not listed. 

779 

780 METHOD 

781 

782 Method/guideline followed: Not stated 

783 

784 Test Type (endpoint): Static (growth rate) 

785 

786 GLP (Y/N) : Yes 

787 

788 Analytical monitoring: No 

789 

790 Year study performed: 2001 

791 

792 Species : Scenedesmus subspicatus 

793 

794 Exposure period : 72 hours 

795 

796 Limit test: Not stated 

797 

798 Dose/Concentration tested : Each concentration (0, 10, 20, 40, 80, 160, 320, 640 and 1000 

799 mg/L) was tested in triplicate . 

800 

801 Remark: 

802 Water media Type: freshwater 

803 Test Concentrations : Nominal 

804 

805 The data came from an English translation of a German language study. 

806 

807 Two stem solutions of the concentrations 1250 mg/L and 800 mg/L were prepared for the main 

808 study. The dose solutions were then prepared out of these through dilution. The dilution series 

809 was prepared through dilution of the stem solution with demineralized water. Concentrations 

810 tested (10-1000 mg/L) were based on the results of a preliminary study. 

811 
812 Scenedesmus subspicatus were procured on agar for the LAUS GmbH from the Institut fuer 

813 Pflanzewissenschaften/depository of algae culture of the University of Goettingen as the strain 

814 culture. The last procurement took place in June 200000. The strain culture was stored in a 

815 refrigerator . 

816 

817 Every 3 days before the start of the preliminary and main tests a batch of algae was taken out of 

818 the stem culture (Ch.B.210600), the algae were apportioned in a culture flask with 50 ml 

819 foreculture medium (Ch.B.11122000). This was incubated for 72 hours with stirring and lighting. 

820 After adjusting the cell number at about 100,000/ml by means of a photometric measurement and 

821 the addition of foreculture medium the culture was ready for the test . 

822 

823 On the day before the start of the test a sufficient number of test vessels were readied. The stem 

824 solution of the dilution series (concentration in the realm of 10 to 1000 mg/L) was prepared and 

825 pipetted into the test vessel (4.0 ml each), such that a mixture of the added foreculture and 

826 nutrient concentrate was achieved (every 0.50 ml, produced 1 .00 ml). Each concentration (0, 10, 

827 20, 40, 80, 160, 320, 640 and 1000 mg/L) was tested in triplicate . 

828 

829 The test vessels were incubated open under stirring (magnet stirrers) at 8000 lux lighting at 21 

830 +/- 2 C. Every 24 hours the cell number was determined by means of a photometric 

831 measurement at 440 nn. Before and after the test the pH values of the control were measured. 

832 The measurement values of the optical density were recorded and evaluated with a computer 

833 program. The measured extinctions and the inherent coloration of the mixture were corrected for 

834 the calculation of the biomass integrals and the growth rate . The growth rates were compared 

835 against the dilution of the sample. The following equation was used for the calculation of the 

836 growth rate . 

837 

838 u = ln(Ext440 nm (72h))- ln(Ext440nm(Oh))/3 

839 

840 The 72h-ErC50 concentration was calculated where the growth rate was inhibited by 50%. The 

841 value is obtained graphically. For this the inhibition values were obtained versus the logarithm of 

842 the concentration. The ErC50 is defined as the intersection of the regression straight line with the 

843 abscissa parallel at 50%. 

844 

845 The area A under the growth curve was calculated for every test culture with the help of the 

846 following equation. 

847 

848 A = (N1-NO/2)*tl + [(N1 + N2 - 2*NOY2](t2-tl)+, . .+(Nn-1 + Nn-2*NO/2)(Tn-tn-1) 

849 

850 Where: 

851 tl : The time of the first measurement 

852 tn: The time of the n-ten measurement 

853 N0: the nominal beginning cell number 

854 N1 : The measured cell number at Time tl 

855 Nn: The measured cell numbe at Time tn 

856 

857 The 72h-EbC50 the concentration was calculated where the growth rate was inhibited by 50%. 
858 The value is obtained graphically. For this the inhibition values were obtained versus the 

859 logarithm of the concentration . The ErC50 is defined as the intersection of the regression straight 

860 line with the abscissa parallel at 50%. 

861 

862 RESULTS 

863 

864 Detailed results: 

865 The results listed above for EC50 and EC 100 are for growth rate. The EC50 and EC 100 values 

866 for biomass are 220 mg/L and > 1000 mg/L, respectively. 

867 

868 The pH variation was 0.5 units. The nominal growth factor that is expected for a control run is 

869 0.9 . The control for the preliminary test had a growth rate of 1.07, and the one for the main test 

87o had a growth rate of 1 .04. 

871 

872 CONCLUSIONS 

873 

874 72 hour NOEC = 10 mg/L 

875 72 hour EC50 = 370 mg/L (growth rate) 

876 72 hour EC 100 > 1000 mg/L (growth rate) 

877 72 hour EC50 = 220 mg/L (biomass) 

878 72 hour EC 100 > 1000 mg/L (biomass) 

879 

88o RELIABILITY CODE 

881 

882 1- Valid Without Restrictions 

883 Flag: Critical study for SIDS endpoint 

884 

885 REFERENCES 

886 

887 Paulus, J. 2001 . Determination of the 72 hour EC50 of "Brueggolit E02" in the presence of 

888 Scenedesmus subspicatus. LAUS-Libertaere Arbeitsgemeinschaft Environmental & Hazard 

889 Substance Consultation GmbH Report No. 00110605J01, dated 2001-02-21 . English translation 

890 of an unpublished German language study.) 

891 

892 Robust Summaries and Dossier for CAS No . 149-44-0, PCA Services, Inc. 

893 

894 

895 

896 

897 

898 

899 

900 

901 

902 

903 
~904 

905 

906 Title: Toxicity to Aquatic Plants e.g. Algae 

907 

908 The study is: ® Adequate 0 Not Adequate 

909 

91o TEST SUBSTANCE 

911 

02 Identity: Methanesulfinic acid, hydroxyl-, monosodium salt (CAS No. 149-44-0) 

913 

914 Remarks: Analytical purity: 100.3% calculated as dihydrate. Batch Number 07041205 from 

915 Bruggermann., The chemical is a commercial product containing sodium 

916 hydroxymethanesulfinate and 1-3% CAS No. 497-19-8 (sodium carbonate) . The exact purity is 

917 unknown. 

918 

919 METHOD 

920 

921 Method/guideline followed: OECD 201 

922 

923 Test Type (endpoint): Static 

924 

925 GLP (Y/N) : Yes 

926 

927 Analytical monitoring: No 

928 

929 Year study performed: 1999 

930 

931 Species: Scenedesmus subspicatus 

932 

933 Exposure period: 72 hours 

934 

935 Limit test: No data 

936 

937 Dose/Concentration tested: 0, 0.1, 1, 10, 100, 500 mg/L 

938 

939 Remark: 

940 Water Media Type: Freshwater 

941 Test Concentrations : Nominal 

942 

943 The data came from an English translation of a German language study. The EC50 value 

944 reported above is for cell number. The EC50 value for growth rate is 450 mg/L 

945 

946 The test organism was Scenedesmus subspicatus (SAG 86.81). The stem culture was inoculated 

947 with a 0.8% agar-agar consolidated, sterile nutrient medium in an inclined agar tubule every two 

948 months. Between inoculations the culture was kept in a cooling cabinet at about 8 C. Before the 

949 inoculation the inclined agar tube was placed 1-2 days in light. 
950 

951 For the preparation of the foreculture, a transverse agar tublet was charged with the stem culture 

952 and placed for 1-2 days in the light, and then inoculated in about 100 mL sterile medium. 

953 Incubation then followed in a shaking machine (100 - 150 rpm) for about 72 hours at 23 +/- 1 C, 

954 and 8.000 lx (without a dark phase) . 

955 

956 Exponentially growing cultures of Scenedesmus subspicatus (10,000 cells/mL) in defined 

957 medium were exposed in triplicate to varying concentrations of test substance (0, 0.1, 1, 10, 100, 

958 500 mg/L) over a time period of 72 hours in a circular shaking machine with standard lighting 

959 attached. The pH for each concentration was determined at 0 and 72 hours. The cell count was 

960 determined by measuring the extinction at 520 nm in each run every 24 hours. The mean value 

961 of the cell number was plotted for every test concentration versus the time, in order to obtain the 

962 growth curves . The inhibition of the cell growth rate and the specific growth rate was 

963 determined in comparison to a control. 

964 

965 RESULTS 

966 

967 Detailed results: 

968 The 72 hour EC50 values for cell number and growth rate were 5 mg/L and 450 mg/L, 

969 respectively . 

970 

971 The test was valid, as algae increased by a factor of 100 in 72 hours (greater than the 16 times 

972 specified by the OECD Guideline) . 

973 

974 CONCLUSIONS 

975 

976 72 hour EC50 = 5 mg/L (cell number) 

977 72 hour EC50 = 450 mg/L (growth rate) 

978 

979 RELIABILITY CODE 

980 

981 1 - Valid Without Restrictions 

982 Flag: Supporting study for SIDS endpoint. 

983 

984 REFERENCES 

985 

986 Jaeger, W. 1999. Algae Inhibition Test According to OECD 201 with Sodium Hydroxy Methane 

987 Sulfinate. Unpublished Report dated 1999-05-12. English translation of a German language 

988 study. 

989 

990 Robust Summaries and Dossier for CAS No. 149-44-0, PCA Services, Inc. 

991 

992 

993 

994 

995 
-996 

997 

998 ACUTE TOXICITY STUDY 

999 

1000 Summary of endpoint: Inadequate 

1001 

1002 Title: Acute Oral Toxicity 

1003 

1004 The study is : F~ Adequate ® Not Adequate 

1005 

1006 This study was missing information such as test substance purity, GLP status, strain of animal 

1007 used, number of animals/sex/dose tested, route of administration, vehicle used, doses 

1008 administered, and method/guideline followed. Based on a weight of evidence, this study was 

1009 given a klimisch code of 2 - valid with restrictions in HPVIS. However, because this study was 

1010 missing critical study details, the study is considered inadequate . 

1011 

1012 TEST SUBSTANCE 

1013 

1014 Identity: Methanesulfinic acid, hydroxyl-, monosodium salt (CAS No. 149-44-0) 

1015 

1016 Remarks: Not stated 

1017 

1018 METHOD 

1019 

1020 Method/guideline followed: Not stated 

1021 

1022 Test Type: Oral 

1023 

1024 GLP (Y/N) : Not stated 

1025 

1026 Year study performed: Not stated 

1027 

1028 Species/strain : Rat 

1029 

1030 Sex: Not stated 

1031 

1032 Number of animals : Not stated 

1033 

1034 Route of administration : Not stated 

1035 

1036 Vehicle: Not stated 

1037 

1038 Doses: Not stated 

1039 

1040 Method remarks: Not stated 

1041 
1042 RESULTS 

1043 

1044 Detailed results: Not stated 

1045 

1046 CONCLUSIONS 

1047 

1048 LD50 > 2 glkg 

1049 

1050 RELIABILITY CODE 

1051 

1052 3 - Not valid 

1053 

1054 REFERENCES 

1055 

1056 Author unknown. 1980. Title not given. Yakkyoku Pharrncy. 31 :959, cited in RTECs. 

1057 Reference ordered. 

1058 

1059 

1060 

1061 

1062 

1063 

1064 

1065 

1066 

1067 

1068 

1069 

1070 

1071 

1072 

1073 

1074 

1075 

1076 

1077 

1078 

1079 

1080 

1081 

1082 

1083 

1084 

1085 

1086 

1087 
1088 

1089 Title: Acute Oral Toxicity 

1090 

1091 The study is : 0 Adequate ® Not Adequate 

1092 

1093 This study was missing information such as test substance purity, GLP status, strain of animal 

1094 used, number of animals/sex/dose tested, route of administration, vehicle used, doses 

1095 administered, and method/guideline followed. Based on a weight of evidence, this study was 

1096 given a klimisch code of 2 - valid with restrictions in HPVIS. However, because this study was 

1097 missing critical study details, the study is considered inadequate . 

1098 

1099 TEST SUBSTANCE 

1100 

1101 Identity: Methanesulfinic acid, hydroxyl-, monosodium salt (CAS No. 149-44-0) 

1102 

1103 Remarks: Not stated 

1104 

1105 METHOD 

1106 

1107 Method/guideline followed: Not stated 

1108 

1109 Test Type: Oral 

1110 

1111 GLP (Y/N) : Not stated 

1112 

1113 Year study performed: Not stated 

1114 

1115 Species/strain : Mouse 

1116 

1117 Sex: Not stated 

1118 

1119 Number of animals: Not stated 

1120 

1121 Route of administration: Not stated 

1122 

1123 Vehicle: Not stated 

1124 

1125 Doses: Not stated 

1126 

1127 Method remarks: Not stated 

1128 

1129 RESULTS 

1130 

1131 Detailed results : Not stated 

1132 

1133 
1134 CONCLUSIONS 

1135 

1136 LD50 = 4 g/kg 

1137 

1138 RELIABILITY CODE 

1139 

1140 3 - Not valid 

1141 

1142 REFERENCES 

1143 

1144 Author unknown. 1980. Title not given. Yakkyoku Pharmcy. 31 :959, as cited in RTECS. 

1145 REFERENCE ORDERED 

1146 

1147 

1148 

1149 

1150 

1151 

1152 

1153 

1154 

1155 

1156 

1157 

1158 

1159 

1160 

1161 

1162 

1163 

1164 

1165 

1166 

1167 

1168 

1169 

1170 

1171 

1172 

1173 

1174 

1175 

1176 

1177 

1178 

1179 
1180 

1181 REPEATED-DOSE TOXICITY 

1182 

1183 Summary of endpoint: Adequate 

1184 

1185 Title: Repeated Dose/Reproductive/Developmental Toxicity 

1186 

1187 The study is : ® Adequate 0 Not Adequate 

1188 

1189 TEST SUBSTANCE 

1190 

1191 Identity: Methanesulfinic acid, hydroxyl-, monosodium salt (CAS No. 149-44-0) 

1192 

1193 Remarks: as prescribed by l .1-1 .4, Analytical purity : 100.3% calculated as dihydrate. Batch 

1194 Number 07041205 from Bruggermann. 

1195 

1196 METHOD 

1197 

1198 Method/guideline followed: OECD 422 

1199 

1200 Study duration: Males were exposed for 30 days (during 2 weeks prior to mating, during 

1201 mating, and to termination) . Females were exposed for 42-53 days (during 2 weeks prior to 

1202 mating, during mating and gestation, up to Day 4 of lactation) . 

1203 

1204 Test type : Combined 28-day repeated dose toxicity study with the reproductive/developmental 

1205 toxicity screening test by oral gavage 

1206 

1207 GLP (Y/N) : Yes 

1208 

1209 Year study performed: 2008 

1210 

1211 Species/strain: Wistar Han Rat 

1212 

1213 Sex: Both M/F 

1214 

1215 Number of animals per dose groups: Four groups of 10/sex 

1216 

1217 Route of administration : Oral gavage 

1218 

1219 Doses tested and frequency: 100, 300, 1000 mg/kg/day 

1220 

1221 Vehicle: Water 

1222 

1223 Post-observation period: Not stated 

1224 

1225 Statistical methods used: Normally distributed data were analyzed using the Dunnett test, based 
1226 on a pooled variance estimate . The Steel test was applied if data were not normally distributed. 

1227 The Fisher Exact test was applied to frequency data. All tests were two-sided, with p < 0.05 

1228 accepted as the lowest level of significance . No statistical analyses were performed on 

1229 histopathological data. 

1230 

1231 Remarks : 

1232 Type: other: combined 28-day repeated dose toxicity study with the reproductive/developmental 

1233 toxicity screening test by oral gavage (OECD 422 and EPA, 40 CFR 799 .9365) 

1234 

1235 Males were exposed for 30 days (during 2 weeks prior to mating, during mating, and to 

1236 termination) . Females were exposed for 42 - 53 days (during 2 weeks prior to mating, during 

1237 mating and gestation, up to Day 4 of lactation) . 

1238 

1239 Control group: yes 

1240 

1241 A limit dose of 1000 mg/kg/day was selected as the high dose level based on data obtained in a 

1242 preliminary range-finding study. The authors reported a no-observable-effect-level (NOAEL) of 

1243 300 mg/kg for general toxicity. 

1244 

1245 Test condition: Animals: Male and virgin female Cr1:WI(Han) (outbred, SPF-Quality) rats were 

1246 obtained from Charles River, Deutschland, Sulzfeld, Germany. They were acclimated at least 5 

1247 days prior to start of treatment. They were housed in rooms maintained at 19.0 - 23 .5 degrees C, 

1248 with a relative humidity of 26 - 95% and a 12-hour light/12 hour dark cycle. there were 15 air 

1249 changes per hour. 

1250 

1251 Prior to mating, animals were housed in groups of 5 animals/sex/cage in Macrolon cages (MIV 

1252 type, height 18 cm) . Females were caged with males on a one-to-one basis during mating. After 

1253 mating, males were housed in their home cage, with a maximum of 5 animals/sex/cage . Females 

1254 were individually housed after mating and were kept with their offspring until termination. 

1255 

1256 Cages were generally supplied with sterilized sawdust as bedding material and paper as cage1257 enrichment. During overnight activity monitoring, animals were housed individually with 

1258 sterilized sawdust bedding material without paper cage-enrichment. 

1259 

1260 Animals were allowed free access to tap water and pelleted rodent diet. Analyses of bedding, 

1261 paper, diet and water did not reveal any findings that were considered to affect study integrity. 

1262 

1263 Test material : Formulations (w/w) were prepared daily, within 4 hours of dosing, and were 

1264 homogenized to a visually acceptable level. No adjustment was made for specific gravity, 

1265 vehicle and/or formulation. The pH of the solutions was determined weekly. The pH of a 10% 

1266 solution was approximately 10%. 

1267 

1268 On September 13, 2007, duplicate samples from each group were analyzed for accuracy. 

1269 Duplicate samples of dosing solutions taken from the bottom, middle and top for the low and 

1270 high dose groups were also analyzed for homogeneity and stability at 0 hours. Duplicate samples 

1271 of these solutions (taken from the middle) were also analyzed for stability at 5 hours. The 
,1272 analytical method used was based on the results of a separate project for the development and 

1273 validation of the analytical method. 

1274 

1275 The pHs of the formulations were between 8.31 - 8.68 (control), 10.12 - 10.17 (low dose), 10.10 

1276 - 10.17 (mid dose) and 9.94 - 10.02 (high dose) . The accuracy of the formulations was 102 -

1277 109% (low dose), 98-103% (mid dose) and 87-105% (high dose). Based on the results it was 

1278 concluded that the formulations were prepared correctly. The low and high dose formulations 

1279 were homogeneous (< = 10% coefficient of variation) . Formulations at the entire range were 

1280 stable for at least 5 hours when stored at room temperature. 

1281 

1282 Study conduct: Animals were approximately 10 weeks old at start of treatment. A health 

1283 examination was performed prior to treatment to ensure that the animals were in a good state of 

1284 health . A total of 40 animals/sex were randomized into four groups of 10/sex according to body 

1285 weight, with all animals within +/- 20% of the sex mean. They were identified by earmark and 

1286 tattoo . 

1287 

1288 Groups of animals were given 0 (water vehicle), 100, 300 or 1000 mg/kg bw test material (at 5 

1289 mL/ kg bw) by oral gavage, once daily,7 days/week up to the day of scheduled termination. A 

1290 limit dose of 1000 mg/kg/day was selected as the high dose level based on data obtained in a 

1291 preliminary range-finding study. Actual dose volumes were calculated according to the latest 

1292 body weight. 

1293 

1294 Following 14 days of exposure, males and females were cohoused (1 :1). Sibling mating was 

1295 avoided. Detection of mating was confirmed by presence of sperm in the vaginal lavage or by the 

1296 appearance of an intravaginal copulatory plug. This day was designated as Day 0 post-coitum. 

1297 Once mating had occurred, the males and females were separated. A maximum of 14 days was 

1298 allowed for mating. Females were allowed to litter normally. Day 1 of lactation was defined as 

1299 the day when a littler was found completed. Females that were littering were left undisturbed. On 

1300 Day 1 of lactation, all pups were individually identified by a tattoo . Examination of maternal 

1301 care revealed no deficiencies. 

1302 

1303 Parental animals were observed for mortality and viability at least twice daily. At least once 

1304 daily, detailed clinical exams were conducted for all animals. This was also performed outside 

1305 the home cage once before treatment and at weekly intervals. These observations were not 

1306 performed when the animals were mating or housed individually . the time of onset, degree and 

1307 duration was recorded. Cage debris of pregnant females was examined for evidence of abortion 

1308 or premature birth. No signs of abortion, premature birth or difficult or prolonged parturition 

1309 were recorded. 

1310 

1311 Functional observations (hearing ability, pupillary reflex, static righting reflex and grip strength 

1312 and a motor activity test) were recorded for the first five mated males/group and the first five 

1313 females/group with live offspring. During the motor activity test, males were caged individually 

1314 and females were caged with their offspring. The males were tested during week 4 of treatment 

1315 and the assigned females were tested during lactation (all before blood sampling). 

1316 

1317 Males and females were weighed on the first day of exposure and weekly thereafter. Mated 
1318 females were weighed on Days 0, 4, 7, 11, 14, 17 and 20 of gestation, and during lactation on 

1319 Days 1 and 4. Food consumption was recorded weekly (with the exception that it was not 

1320 recorded during breeding). Food consumption of mated females was measured on Days 0, 4, 7, 

1321 11, 14, 17 and 20 post-coiturn and after delivery on Days 1 and 4 post-partum. Water 

1322 consumption was not quantified, but was monitored subjectively. Identities of animals paired, 

1323 mating date, confirmation of pregnancy, and delivery day were recorded. 

1324 

1325 After an overnight fast, blood was collected from the retro-orbital sinuses of the first five mated 

1326 males/group and the first five females/group with live offspring, under iso-flurane anesthesia. 

1327 Standard hematological (white blood cells, differential leukocyte count, red blood cells, 

1328 reticulocytes, red blood cell distribution width, hemoglobin, hematocrit, mean corpuscular 

1329 volume, meancorpuscular hemoglobin, mean corpuscular hemoglobin concentration, platelets, 

1330 prothrombin time, and activated partial thromboplastin time) and clinical biochemistry 

1331 parameters(alanine aminotransferase, aspartate animotransferase, alkaline phosphatase, total 

1332 protein, albumin, total bilirubin, urea, creatinine, glucose, cholesterol, sodium, potassium, 

1333 chloride, calcium, and inorganic phosphate) were preformed after appropriate blood processing. 

1334 

1335 On lactation Day 5, parental females were anesthetized with iso-flurane and exsanguinated. 

1336 Animals were subjected to a gross examination, organ weights (adrenal glands, brain, heart, 

1337 kidneys, liver, spleen, thymus) of the first 5 females with live offspring per group were collected 

1338 and terminal body weights were recorded. Tissues (adrenal glands, aorta, brain, cecum, cervix, 

1339 clitoral gland, colon, duodenum, eyes with optic nerve and Harderian gland, mammary gland 

1340 area, femur, heart, ileum, kidneys, larynx, exorbital lacrimal gland, liver, lung, mandibular and 

1341 mesenteric lymph nodes, nasopharynx, esophagus, ovaries, pancreas, Peyer's patches, pituitary 

1342 gland, preputial gland, rectum, mandibular and sublingual salivary glands, sciatic nerve, skeletal 

1343 muscle, skin, spinal cord, spleen, sternum with bone marrow, stomach, thymus, thyroid with 

1344 parathyroid, tongue trachea, urinary bladder) and gross lesions of these animals were preserved 

1345 for possible future histopathological examination. The cervix, clitoral gland, ovaries, preputial 

1346 gland, uterus, vagina and all gross lesions from remaining animals also were preserved. Slides of 

1347 preserved organs and tissues of selected control and high dose animals and gross lesions of all 

1348 animals were prepared and examined by a pathologist. The numbers of former implantation sites 

1349 and corpora lutea were counted and recorded. 

1350 

1351 On post-mating Days 24-26, females that had not delivered were anesthetized and exsanguinated. 

1352 No organs were collected for weighing. Animals were subjected to a gross examination and 

1353 tissues described above for parental females (and any gross lesions) were preserved. The 

1354 reproductive organs of all females that failed to conceive were examained by a pathologist. 

1355 Additionally, nongravid uteri were stained using the Salweski technique to identify former 

1356 implantation sites. 

1357 

1358 After completion of the mating period, parental males were anesthetized with iso-flurane and 

1359 exsanguinated and grossly examined. Organ weights of the first 5 mated males per group were 

1360 collected and terminal body weights were recorded. Weights of epididymides and testes of all 

1361 males were also recorded. The same tissues preserved for selected females (see above) were 

1362 preserved for the first 5 mated males per group (with the exception of female reproductive 

1363 organs and the addition of the epididymides, prostate gland, seminal vesicles, testes, and 
1364 coagulating gland). The coagulation gland, epididymides, preputial gland, prostate gland, 

1365 seminal vesicles, testes and all gross lesions from remaining animals also were preserved. Slides 

1366 of preserved organs and tissues of selected control and high dose animals and gross lesions of all 

1367 animals were prepared and examined by a pathologist. Of the selected 5 males/group of the 

1368 control and high dose group, additional slides of the testes were prepared to examine staging of 

1369 spermatogenesis. The reproductive organs of all males that failed to sire were examined by a 

1370 pathologist. 

1371 

1372 Statistical analyses : Normally distributed data were analyzed using the Dunnett test, based on a 

1373 pooled variance estimate. The Steel test was applied if data were not normally distributed . The 

1374 Fisher Exact test was applied to frequency data. All tests were two-sided, with p < 0.05 accepted 

1375 as the lowest level of significance . No statistical analyses were performed on histopathological 

1376 data. 

1377 

1378 Methods pertaining to reproductive toxicity and offspring are presented in Sections 5.8 .1 and 

1379 5 .8 .2 . 

1380 

1381 RESULTS 

1382 

1383 NOAEL = 300 mg/kg bw/day 

1384 

1385 LOAEL =1000 mg/kg bw/day 

1386 

1387 

1388 

1389 

1390 

1391 

1392 

1393 

1394 

1395 

1396 

1397 

1398 

1399 

1400 

1401 

1402 

1403 

1404 

1405 

1406 

1407 

1408 

1409 

Remarks: 

Parental animals: 

1000 mg/kg-bw/day: None of the animals died and functional observations were not affected. 

One female showed hunched posture and piloerection from Day 20 post-coitum until Day 3 of 

lactation. Piloerection was noted in another female on Day 4 of lactation. Slight salivation noted 

incidentally in both sexes was attributed to the test of the formulation. Lower mean body weights 

were recorded for males and females at 1000 mg/kg during the complete treatment prior (not 

always statistically significant) . Body weight gains of males were also lower than control 

throughout the treatment period . Food consumption before or after allowance for body weight 

was reduced in males and females during the whole observation period (not always statistically 

significant) . A slightly higher white blood cell count was noted in males (not statistically 

significant) and females. Relative numbers of reticulocytes were increased in males (3 .6 +/-

0.8% in treated vs. 2.3 +/- 0.1 % in control). Increased mean corpuscular volume and decreased 

mean corpuscular hemoglobin concentration in females were not considered to be related to 

treatment as they were either within the range of normal or showed no dose-response 

relationship . Incresed alanine aminotransferase was found in males (59.2 +/- 20.4 U/L in treated 

vs 38 .0 +/- 3 .6 U/L in control) and two females. One male had increased activity of aspartate 

aminotransferase. Increased total bilirubin and decreased sodium in males were not considered 

to be related to treatment due to the lack of a dose-response relationship . One male showed 

exophthalmus of the eye, which was considered to be caused by blood sampling. Increases in 

organ/body weight ratios for the liver (2.57 +/- 0.14% in treated vs. 2.33 +/- 0.12% in control), 

spleen (0.225 +/- 0.032 in treated vs 0.162 +/- 0.033 in control), testes (1 .16 +/- 0.15% in treated 

vs 0.94 +/- 0.16% in control) and epididymides (0.388 +/- 0.071% in treated vs . 0.301 +/- 0/050 
1410 in control) of males and liver (3.64 +/- 0.22% in treated vs. 3 .04 +/- 0.33% in control), spleen 

1411 (0.287 +/- 0.023% in treated vs. 0.225 +/- 0.0303% in control), kidneys (0.83 +/- 0.05% in 

1412 treated vs. 0.72 +/- 0.06% in control) and adrenals (0.043 +/- 0.004% in treated vs. 0.034 +/- 

1413 0.004% in control) of females were observed. There were no corresponding histological changes. 

1414 an increase in the organ/body weight ratio of the brain in males (0.66 +/- 0.05% in treated vs. 

1415 0.60 +/- 0.01% in control) was considered to be due to the relatively low terminal body weight 

1416 (300 +/- 14 g in treated vs. 336 +/- 19 g in control). Absolute brain weight (but not relative) was 

1417 slightly decreased in females (1 .77 +/- 0.05 g in treated vs. 1 .87 +/- 0.05 in control). 

1418 Reproductive and breeding parameters were not affected by treatment with up to 1000 

1419 mg/kg/day. 

1420 

1421 300 mg/kg bw/day: None of the animals died and functional observations were not affected. 

1422 Females had slightly lower mean absolute body weights on Day 8 of premating (189 +/- 6.6 g in 

1423 treated vs. 197 +/- 7.2 g in control). As this had recovered on Day 1 of mating (198 +/- 11 .8 g in 

1424 treated vs. 203 +/- 8.2 g in control), it was not considered to be toxicologically relevant. 

1425 Increased basophils in males, increased mean corpuscular volume and mean corpuscular 

1426 hemoglobin in females, decreased total bilirubin in males, and decreased albumin in females 

1427 were not considered to be related to treatment as they were either within the range of normal or 

1428 showed no dose-response relationship . No other abnormal findings were noted. 

1429 

1430 100 mg/kg bw/day: None of the animals died and functional observations were not affected. 1431 Females had slightly lower mean absolute body weights on Day 8 of premating (189 +/- 6 .6 g in 

1432 treated vs 197 +/- 7.2 g in control). As this had recovered on Day 1 of mating (196 +/- 6.3 g in 

1433 treated vs. 203 +/- 8 .2 g in control), it was not considered to be toxicologically relevant. 

1434 Increased basophils in males and decreased total bilirubin in males were not considered to be 

1435 related to treatment as they were either within the range of normal or showed no dose-response 

1436 relationship. No other abnormal findings were noted. 

1437 

1438 Control: one female showed exophthalmus of the eye, which was considered to be cause by 

1439 blood sampling. 

1440 

1441 Results pertaining to reproductive toxicity and offspring are presented in sections 5.8 .1 and 5.8 .2 . 

1442 

1443 CONCLUSIONS 

1444 

1445 NOAEL = 300 mg/kg bw/day 

1446 

1447 LOAEL =1000 mg/kg bw/day 

1448 

1449 RELIABILITY CODE 

1450 . 

1451 1 - Valid without restrictions 

1452 Flag: Critical study for SIDS endpoint 

1453 

1454 

1455 
1456 REFERENCE 

1457 

145s Beekhuijzen, M.E.W. 2008 . A combined 28-day repeated dose toxicity study with the 

1459 reproduction/developmental toxicity screening test of Methane sulfinic acid, hydroxy-, 

1460 monosodium salt in rats by oral gavage. NOTOX B.V. Report No. 485218 (485615), dated 01-

1461 15-2008 (first draft of an unpublished study report) 

1462 

1463 Robust Summaries and Dossier for CAS No. 149-44-0, PCA Services, Inc. 
1464 GENETIC TOXICITY - GENE MUTATIONS 

1465 

1466 Summary of endpoint: Inadequate 

1467 

1468 Title: Genetic Toxicity 

1469 

1470 The study is : [:] Adequate ® Not Adequate 

1471 

1472 This was not a guideline study. It provided far too little detail to know whether the test chemical 

1473 induces gene mutations, although the sex-linked recessive lethal part of the experiment suggests 

1474 that it might do so. Serious deficiencies were the lack of many necessary details, of positive and 

1475 (in some cases) solvent or negative controls, and of statistical analysis. 

1476 

1477 TEST SUBSTANCE 

1478 

1479 Identity: Methanesulfinic acid, hydroxyl-, monosodium salt (CAS No. 149-44-0) 

1480 

1481 Remarks : No indication of the purity was provided. 

1482 

1483 METHOD 

1484 

1485 Method/guideline followed: Not stated 

1486 

1487 Year Study Performed: 1969 

1488 

1489 GLP: No Data 

1490 

1491 Test type: Sex-linked recessive lethal test in Drosophila 

1492 

1493 Species/strain/cell type or line: Drosophila 

1494 

1495 Sex: Male 

1496 

1497 Number of animals/sex/dose : Not stated 

1498 

1499 Route of administration : Intraperitoneal 

1500 

1501 Vehicle: Yes 

1502 

1503 Doses: Flies were injected intraperitoneally with 0.0004 ml of 0.06 M test chemical in a 0.5% 

1504 NaCl solution . This is equivalent to 2.8 gg of the test chemical per fly. 

1505 

1506 Method remarks: 

1507 Frequency of treatment: 1 

1508 Positive, Negative and solvent Control Substance(s): Positive Control Substance Remarks : None 

1509 
i51o Negative Control Substance Remarks: There was no concurrent solvent or negative control for 

1511 the sex-linked recessive lethal mutation test or the translocation test . (Because of the slight 

1512 information content of that part of the study, no Robust Study Summary is being prepared for the 

1513 translocation part of the experiment.) 

1514 

1515 Method/Guideline and Test Condition Remarks: There should have been a positive control. 

1516 While they did have concurrent controls for the experiments involving chromosomes 2 and 3, 

1517 there was no concurrent solvent or negative control for the sex-linked recessive lethal test . 

1518 Instead, for comparison they used a point estimate of a spontaneous mutation rate that 

1519 presumably came from the literature . It is unknown if their concurrent controls were solvent 

1520 controls or negative controls. According to the OECD TG 477, it is recommended that injections 

1521 be dissolved in 0.7% saline . Instead the present study used 0.5% saline. The guideline 

1522 recommends injecting males when they are 3 to 5 days old. Instead the present study injected 

1523 males when they were one day old. Details should have been provided regarding how they made 

1524 certain that their wild-type strain was lethal free. There was also no mention of testing to ensure 

1525 that mutations that were thought to be induced were not instead part of a cluster of spontaneous 

1526 mutations, which is a potentially serious complication (Woodruff, R.C. et al ., 1996. Clusters of 

1527 identical new mutation in the evolutionary landscape. Genetica 98 :149-160.) There was no 

1528 statistical analysis. This was a less than one-page report, and it did not contain enough detail to 

1529 support its claim that the test substance is weakly mutagenic in the fly for lethal mutations. 

1530 

1531 Method/Guideline Description: The sex-linked recessive lethal part of the test has some 

1532 similarity to OECD TG 477. Wild-type Oregon-R males of the Oak Ridge strain were made free 

1533 of pre-existing lethals in the X, second and third chromosomes by an unspecified procedure. 

1534 Newly-hatched males were collected within a two-hour hatching period and were aged one day 

1535 before intraperitoneal injection. The treated males were mated with Ins y^3p;Cy/Pm;D, 

1536 InCXF/Sb females one day after injection. The use of this test stock was said to permit 

1537 simultaneous testing for lethal mutations on any of the chromosomes designated X, 2, and 3, as 

1538 well as for translocations involving chromosomes 2 or 3 or Y. They also used a 3-day brood 

1539 technique over a 16-day test period to test for sex-linked recessive lethals. There was no 

1540 indication of using a brood technique for autosomal recessive lethals . They also gave 

1541 experimental results for lethals expressed as mosaics in the F3, but no control rate was presented 

1542 for comparison. 

1543 

1544 Vehicle used: Yes 

1545 Vehicle Name: Other 

1546 

1547 Species : Drosophila melanogaster 

1548 

1549 RESULTS 

1550 

1551 Genotoxic Effect: Equivocal 

1552 

1553 Remarks: If the flies used in the sex-linked lethal test were indeed lethal free at the beginning of 

1554 the experiment, then that part of the experiment showed that there was induction of sex-linked 

1555 recessive lethal mutations, especially in the first brood and probably continuing for a few more 
1556 broods. This conclusion relies upon a comparison with an apparently literature-based control 

1557 mutation rate said to exceed 0.04% rarely . In the first brood, they reported a mutation rate of 

1558 8/1095 gametes or 0.75% . The authors also claim to have demonstrated induction of lethal 

1559 mutations in chromosomes 2 and 3. In this instance, however, they presented the actual data for 

1560 both the experimental and control groups, and the reviewer could thus evaluate them statistically. 

1561 None of the comparisons approached statistical significance . The smallest P-value for a one1562 tailed Fishers Exact Test was 0.25 . Even if the results for both chromosomes 2 and 3 are 

1563 combined, P = 0.12. They claimed that this effect (which is shown here to be not statistically 

1564 significant) was twice as large as the effect that they demonstrated on the X chromosome. No 

1565 translocations were found in analysis of 1154 mature sperm. 

1566 

1567 Strain text description: Oregon-R Oak Ridge strain 

1568 

1569 Criteria for Postive Results : Not provided; should be a statistically significant increase over 

1570 concurrent solvent controls 

1571 

1572 CONCLUSION 

1573 

1574 The results for the sex-linked recessive lethal test are considered equivocal since there were 

1575 serious shortcomings in the method and so few details were provided. The results for induction 

1576 of recessive lethal mutations in chromosomes 2 and 3 and translocations are also considered 

1577 equivocal. While no evidence of induction of lethal mutations on those two chromosomes (or of 

1578 translocations) was found, it cannot be concluded that the test substance does not induce such 

1579 mutations because the samples were too small, there was no positive control, and it is unknown if 

1580 the dose was as high as it should have been. 

1581 

1582 RELIABILITY CODE 

1583 . 

1584 3-Not Valid 

1585 This was not a guideline study. It provided far too little detail to know whether the test chemical 

1586 induces gene mutations, although the sex-linked recessive lethal part of the experiment suggests 

1587 that it might do so. Serious deficiencies were the lack of many necessary details, of positive and 

1588 (in some cases) solvent or negative controls, and of statistical analysis. 

1589 Key study sponsor indicator : Not Key 

1590 

1591 REFERENCE 

1592 

1593 Paik, Y.K. and Y. Choi. 1969. The mutagenic effect of formaldehyde sodium sulfoxylate on 

1594 male germ cells of D. melanogaster. Drosophila Information Service. 44:69. 

1595 

1596 

1597 

1598 

1599 

1600 

1601 
1602 

1603 

1604 

1605 

1606 

1607 

1608 

1609 

1610 

1611 

1612 

1613 

1614 

1615 

1616 

1617 

1618 

1619 

1620 

1621 

1622 

1623 

1624 

1625 

1626 

1627 

1628 

1629 

1630 

1631 

1632 

1633 

1634 

1635 

1636 

1637 

1638 

1639 

1640 

1641 

1642 

1643 

1644 

1645 

1646 

1647 

GENETIC TOXICITY - CHROMOSOMAL ABERRATIONS 

Summary of endpoint: Adequate 

Title: Genetic Toxicity 

The study is : ® Adequate ~ Not Adequate 

TEST SUBSTANCE 

Identity: Methanesulfinic acid, hydroxyl-, monosodium salt (CAS No. 149-44-0) 

Remarks: Batch 07041205 of Methane Sulfinic Acid, Hydroxy-, Monosodium Salt was a white 

powder of a crystalline substance consisting of colourless crystals with a purity of 100.3%. 

METHOD 

Method/guideline followed: OECD 474 

Year Study Performed: 2008 

GLP: Yes 

Test type: Bone Marrow Chromosomal Aberration, Micronucleus assay 

Species/strain/cell type or line : NMRI Mouse 

Sex: Both M/F 

Number of animals/sex/dose : Five male and five female animals were used in each of the six 

treatment groups 

Route of administration : Intraperitoneal 

Vehicle: physiological saline 

Doses: Animals were dosed with Methane Sulfinic Acid, Hydroxy-, Monosodium Salt at 2000 

(two groups), 1000 (one group), and 500 (one group) mg/kg body weight. 

Method remarks: 

Frequency of treatment: 1 x 

Positive, Negative and Solvent Control Substance(s) : The negative and positive control groups 

were treated with vehicle and 50 mg/kg body weight of cyclophosphamide (CP), respectively . 

Bone marrow of the negative and positive control groups was harvested 24 and 48 hours after 

dosing, respectively. 
1648 

1649 Post-Exposure Period: 24 or 48 (highest dose only) hours 

1650 

1651 Number of Animals per Sex per Dose: Five male and five female animals were used in each of 

1652 the six treatment groups, including negative and positive controls. 

1653 

1654 Method/Guideline and Test condition Remarks: The study procedures described in this report 

1655 were based on the most recent OECD, EEC and EPA guidelines. 

1656 

1657 The test substance was dissolved in physiological saline . 

1658 Five male and five female animals were used in each of the six treatment groups, including 

1659 negative and positive controls. All groups received a single intraperitoneal injection. The 

1660 negative and positive control groups were treated with vehicle and 50 mg/kg body weight of 

1661 cyclophosphamide (CP), respectively. Animals were dosed with Methane Sulfinic Acid, 

1662 Hydroxy-, Monosodium Salt at 2000 (two groups), 1000 (one group), and 500 (one group) 

1663 mg/kg body weight. All animals showed no abnormalities after dosing. 

1664 

1665 Bone marrow of the groups treated with Methane Sulfinic Acid, Hydroxy-, Monosodium Salt 

1666 was sampled 24 or 48 (highest dose only) hours after dosing . Bone marrow of the negative and 

1667 positive control groups was harvested 24 and 48 hours after dosing, respectively. 

1668 

1669 RESULTS 

1670 

1671 Systemic Toxicity: No effects 

1672 

1673 Genotoxic Effect: Positive 

1674 

1675 Remarks: Methane Sulfinic Acid, Hydroxy-, Monosodium Salt induced a statistically significant 

1676 increase in the mean number of micronucleated polychromatic erythrocytes compared to the 

1677 corresponding solvent control group in male and female animals dosed with 2000, 1000 and 500 

1678 mg/kg body weight at the 24 hours sampling time. The number of micronucleated polychromatic 

1679 erythrocytes in the bone marrow of all animals treated Methane Sulfinic Acid, Hydroxy-, 

1680 Monosodium Salt and sampled after 24 hours was above the historical control data range. 

1681 

1682 Therefore the increases are biologically relevant and Methane Sulfinic Acid, Hydroxy1683 ,Monosodium Salt is considered clastogenic. 

1684 

1685 Methane Sulfinic Acid, Hydroxy-, Monosodium Salt did not induce a statistically significant 

1686 increase in the mean number of micronucleated polychromatic erythrocytes compared to the 

1687 corresponding solvent control group in male and female animals dosed with 2000 mg/kg body 

1688 weight at the 48 hours sampling time. 

1689 

1690 The incidence of micronucleated polychromatic erythrocytes in the bone marrow of all negative 

1691 control animals was within the historical solvent control data range. Cyclophosphamide, the 

1692 positive control substance, induced a statistically significant increase in the number of 

1693 micronucleated polychromatic erythrocytes in both sexes. Hence, both criteria for an acceptable 
1694 assay were met. 

1695 

1696 The groups that were treated with Methane Sulfinic Acid, Hydroxy-, Monosodium Salt showed 

1697 no decrease in the ratio of polychromatic to normochromatic erythrocytes compared to the 

1698 vehicle controls, which reflects a lack of toxic effects of this compound on the erythropoiesis . 

1699 

1700 The groups that were treated with cyclophosphamide showed an expected decrease in the ratio of 

1701 polychromatic to normochromatic erythrocytes compared to the vehicle controls, demonstrating 

1702 toxic effects on erythropoiesis . 

1703 

1704 CONCLUSION 

1705 

1706 It is concluded that Methane Sulfinic Acid, Hydroxy-, Monosodium Salt is clastogenic in the 

1707 micronucleus test under the experimental conditions described in this report. 

1708 

17o9 RELIABILITY CODE 

1710 

1711 1 - Valid Without Restrictions 

1712 

1713 REFERENCE 

1714 

1715 Study Title: MICRONUCLEUS TEST IN BONE MARROW CELLS OF THE MOUSE WITH 

1716 METHANE SULFINIC ACID, HYDROXY-, MONOSODIUM SALT Author: Drs. C.A.F. 

1717 Buskens, Study completion date: 22 May 2008, NOTOX B.V., NOTOX Project 485472 

1718 

1719 The robust summary for this study was also provided in: Robust Summaries and Dossier for 

1720 CAS No. 149-44-0, PCA Services, Inc. 

1721 

1722 

1723 

1724 

1725 

1726 

1727 

1728 

1729 

1730 

1731 

1732 

1733 

1734 

1735 

1736 

1737 

1738 

1739 
1740 REPRODUCTIVE TOXICITY 

1741 

1742 Summary of endpoint: Adequate 

1743 

1744 Title: Repeated-dose/reproductive/developmental toxicity study 

1745 

1746 The study is: ® Adequate F] Not Adequate 

1747 

1748 TEST SUBSTANCE 

1749 

1750 Identity: Methanesulfinic acid, hydroxyl-, monosodium salt (CAS No. 149-44-0) 

1751 

1752 Remarks: as prescribed by 1 .1-1 .4, Analytical purity : 100.3% calculated as dihydrate. Batch 

1753 Number 07041205 from Bruggemann. 

1754 

1755 METHOD 

1756 

1757 Method/guideline followed: OECD 422 

1758 

1759 Study duration: Males were exposed for 30 days (during 2 weeks prior to mating, during 

1760 mating, and to termination) . Females were exposed for 42-53 days (during 2 weeks prior to 

1761 mating, during mating and gestation, up to Day 4 of lactation) . 

1762 

1763 Test type: combined 28-day repeated dose toxicity study with the reproductive/developmental 

1764 toxicity screening test by oral gavage 

1765 

1766 GLP (Y/N) : Yes 

1767 

1768 Year study performed: 2008 

1769 

1770 Species/strain: Wistar Rat 

1771 

1772 Sex: both M/F 

1773 

1774 Number of animals per dose groups: four groups of 10/sex 

1775 

1776 Route of administration : oral gavage 

1777 

1778 Doses tested and frequency: 100, 300, 1000 mg/kg/day 

1779 

1780 Vehicle : water 

1781 

1782 Post-observation period: Not stated 

1783 

1784 Statistical methods used: Normally distributed data were analyzed using the Dunnett test, based 

1785 on a pooled variance estimate . The Steel test was applied if data were not normally distributed. 
1786 The Fisher Exact test was applied to frequency data. All tests were two-sided, with p < 0.05 

1787 accepted as the lowest level of significance. No statistical analyses were performed on 

1788 histopathological data. 

1789 

1790 Remarks: 

1791 Frequency of Treatment : daily 

1792 

1793 Method/Guideline and Test Condition Remarks: Type: other: combined 28-day repeated dose 

1794 toxicity study with the reproductive/developmental toxicity screening test by oral gavage (OECD 

1795 422 and EPA, 40 CFR 799.9365) 

1796 

1797 Males were exposed for 30 days (during 2 weeks prior to mating, during mating, and to 

1798 termination). Females were exposed for 42-53 days (during 2 weeks prior to mating, during 

1799 mating and gestation, up to Day 4 of lactation). 

1800 

1801 Control group : yes 

1802 

1803 Test condition: Animals: Male and virgin female Cr1:W1(Han) (outbred, SPF-Quality) ratswere 

1804 obtained from Charles River, Deutschland, Suizfeld, Germany. They were acclimated at least 5 

1805 days prior to start of treatment. They were housed in rooms maintained at 19.0 - 23 .5 degrees C, 

1806 with a relative humidity of 26 - 95% and a 12-hour light/12 hour dark cycle. There were 

1807 15 air changes per hour. Prior to mating, animals were housed in groups of 5 animals/sex/cage in 

1808 Macrolon cages (MIV type, height 18 cm). Females were caged with males on a one-to-one basis 

1809 during mating. After mating, males were housed in their home cage, with a maximum of 5 

1810 animals/sex/cage. Females were individually housed after mating and were kept with their 

1811 offspring until termination. Cages were generally supplied with sterilized sawdust as bedding 

1812 material and paper as cage-enrichment. During overnight activity monitoring, animals were 

1813 housed individually with sterilized sawdust bedding material without paper cage-enrichment. 

1814 Animals were allowed free access to tap water and pelleted rodent diet (SM RIM-Z from 

1815 SSNIFF Spezialdiaten GmbH, Soest, Germany). Analyses of bedding, paper, diet and water did 

1816 not reveal any findings that were considered to affect study integrity. 

1817 

1818 Test material : Formulations (w/w) were prepared daily, within 4 hours of dosing, and were 

1819 homogenized to a visually acceptable level. No adjustment was made for specific gravity, 

1820 vehicle and/or formulation. The pH of the solutions was determined weekly. The pH of a 10% 

1821 solution was approximately 10% On September 13, 2007, duplicate samples from each group 

1822 were analyzed for accuracy. Duplicate samples of dosing solutions taken from the bottom, 

1823 middle and top for the low and high dose groups were also analyzed for homogeneity and 

1824 stability at 0 hours. Duplicate samples of these solutions (taken from the middle) were also 

1825 analyzed for stability at 5 hours . The analytical method used was based on the results of a 

1826 separate project for the development and validation of the analytical method (NOTOX Project 

1827 485209). 

1828 

1829 The pHs of the formulations were between 8 .31 - 8.68 (control), 10.12 - 10.17 (low dose), 10.10 

1830 - 10.17 (mid dose) and 9.94 - 10.02 (high dose). The accuracy of the formulations was 102%-

1831 109% (low dose), 98%-103% (mid dose) and 87% - 105% (high dose). Based on the results, it 
1832 was concluded that the formulations were prepared correctly. The low and high dose 

1833 formulations were homogeneous (<= 10% coefficient of variation) . Formulations at the entire 

1834 range were stable for at least 5 hours when stored at room temperature. Study conduct: Animals 

1835 were approximately 10 weeks old at start of treatment. A health examination was performed 

1836 prior to treatment to ensure that the animals were in a good state of health. A total of 40 

1837 animals/sex were randomized into four groups of 10/sex according to body weight, with all 

1838 animals within +/- 20% of the sex mean. They were identified by earmark and tattoo . 

1839 

1840 Groups of animals were given 0 (water vehicle), 100, 300 or 1000 mg/kg bw test material (at 5 

1841 ml/kg bw) by oral gavage, once daily, 7 days/week up to the day of scheduled termination. A 

1842 limit dose of 1000 mg/kg/day was selected as the high dose level based on data obtained in a 

1843 preliminary range-finding study. Actual dose volumes were calculated according to the latest 

1844 body weight. 

1845 

1846 Following 14 days of exposure, males and females were cohoused (13). Sibling mating was 

1847 avoided. Detection of mating was confirmed by presence of sperm in the vaginal lavage or by the 

1848 appearance of an intravaginal copulatory plug. This day was designated as Day 0 postcoitum. 

1849 Once mating had occurred, the males and females were separated. A maximum of 14 days was 

1850 allowed for mating. Females were allowed to litter normally. Day 1 of lactation was defined as 

1851 the day when a litter was found completed. Females that were littering were left undisturbed. On 

1852 Day 1 of lactation, 1 :1 pups were individually identified by a tattoo . Examination of maternal 

1853 care revealed no deficiencies . 

1854 

1855 Parental animals were observed for mortality and viability at least twice daily. At least once 

1856 daily, detailed clinical exams were conducted for all animals. This was also performed outside 

1857 the home cage once before treatment and at weekly intervals. These observations were not 

1858 performed when the animals were mating or housed individually. The time of onset, degree and 

1859 duration was recorded. Cage debris of pregnant females was examined for evidence of abortion 

1860 or premature birth. No signs of abortion, premature birth or difficult or prolonged parturition 

1861 were recorded. 

1862 

1863 Functional observations (hearing ability, pupillary reflex, static righting reflex and grip strength 

1864 and a motor activity test) were recorded for the first five mated males/group and the first five 

1865 females/group with live offspring. During the motor activity test, males were caged individually 

1866 and females were caged with their offspring. The males were tested during week 4 of treatment 

1867 and the assigned females were tested during lactation (all before blood sampling). 

1868 

1869 Males and females were weighed on the first day of exposure and weekly thereafter . Mated 

1870 females were weighed on Days 0, 4, 7, 11, 14, 17 and 20 of gestation, and during lactation on 

1871 Days 1 and 4. Food consumption was recorded weekly (with the exception that it was not 

1872 recorded during breeding). Food consumption of mated females was measured on Days 0, 4, 7, 

1873 11, 14, 17 and 20 post-coiturn and after delivery on Days 1 and 4 postpartum. Water 

1874 consumption was not quantified, but was monitored subjectively. Identities of animals paired, 

1875 mating date, confirmation of pregnancy, and delivery day were recorded. After an overnight fast, 

1876 blood was collected from the retro-orbital sinuses of the first five mated males/group and the first 

1877 five females/group with live offspring, under iso-flurane anesthesia. Standard hematological 
1878 (white blood cells, differential leukocyte count, red blood cells, reticulocytes, red blood cell 

1879 distribution width, hemoglobin, hematocrit, mean corpuscular volume, mean corpuscular 

1880 hemoglobin, mean corpuscular hemoglobin concentration, platelets, prothrombin time, and 

1881 activated partial thromboplastin time) and clinical biochemistry parameters (alanine 

1882 aminotransferase, aspartate aminotransferase, alkaline phosphatase, total protein, albumin, total 

1883 bilirubin, urea, creatinine, glucose, cholesterol, sodium, potassium, chloride, calcium, and 

1884 inorganic phosphate) were performed after appropriate blood processing. 

1885 

1886 The numbers of live and dead pups at the first litter check (Day 1 of lactation) and daily 

1887 thereafter were determined. Detailed clinical observations of pups were made at least once daily. 

1888 Live pups were weighed during lactation Days 1 and 4. Sex of all pups was determined on Days 

1889 1 and 4 of lactation (by measuring the ano-genital distance). On lactation Day 5 (or shortly 

1890 thereafter), parental females were anesthetized with iso-flurane and exsanguinated. Animals were 

1891 subjected to a gross examination, organ weights (adrenal glands, brain, heart, kidneys, liver, 

1892 spleen, thymus) of the first 5 females with live offspring per group were collected and terminal 

1893 body weights were recorded. Tissues (adrenal glands, aorta, brain, cecum, cervix, clitoral gland, 

1894 colon, duodenum, eyes with optic nerve and Harderian gland, mammary gland area, femur, heart, 

1895 ileum, jejunum, kidneys, larynx, exorbital lacrimal gland, liver, lung, mandibular and mesenteric 

1896 lymph nodes, nasopharynx, esophagus, ovaries, pancreas, Peyer's patches, pituitary gland, 

1897 preputial gland, rectum, mandibular and sublingual salivary glands, sciatic nerve, skeletal 

1898 muscle, skin, spinal cord, spleen, sternum with bone marrow, stomach, thymus, thyroid with 

1899 parathyroid, tongue, trachea, urinary bladder) and gross lesions of these animals were preserved 

1900 for possible future histopathological examination. The cervix, clitoral gland, ovaries, preputial 

1901 gland, uterus, vagina and all gross lesions from remaining animals also were preserved. Slides of 

1902 preserved organs and tissues of selected control and high dose animals and gross lesions of all 

1903 animals were prepared and examined by a pathologist. The numbers of former implantation sites 

1904 and corpora lutea were counted and recorded. 

1905 

1906 On post-mating Days 24-26, females that had not delivered were anesthetized and exsanguinated. 

1907 No organs were collected for weighing. Animals were subjected to a gross examination and 

1908 tissues described above for parental females (and any gross lesions) were preserved. The 

1909 reproductive organs of all females that failed to conceive were examined by a pathologist. 

191o Additionally, nongravid uteri were stained using the Salewski technique to identify former 

1911 implantation sites . After completion of the mating period, parental males were anesthetized with 

1912 iso-flurane and exsanguinated and grossly examined. Organ weights (adrenal glands, brain, 

1913 heart, kidneys, liver, spleen and thymus) of the first 5 mated males per group were collected and 

1914 terminal body weights were recorded. Weights of epididymides and testes of all males were also 

1915 recorded. The same tissues preserved for selected females (see above) were preserved for the 

1916 first 5 mated males per group (with the exception of female reproductive organs and the addition 

1917 of the epididymides, prostate gland, seminal vesicles, testes, and coagulating gland) . The 

1918 coagulation gland, epididymides, preputial gland, prostate gland, seminal vesicles, testes and all 

1919 gross lesions from remaining animals also were preserved. Slides of preserved organs and tissues 

1920 of selected control and high dose animals and gross lesions of all animals were prepared and 

1921 examined by a pathologist. Of the selected 5 males/group of the control and high dose group, 

1922 additional slides of the testes were prepared to examine staging of spermatogenesis. The 

1923 reproductive organs of all males that failed to sire were examined by a pathologist. Pups were 
1924 killed by decapitation on Day 5 of lactation (or shortly thereafter). 

1925 

1926 All offspring were sexed and externally examined. Stomachs were examined for the presence of 

1927 milk. Descriptions of all external abnormalities were recorded. If possible, defects or causes of 

1928 death were evaluated. Any abnormal pup, organ or tissue was preserved for possible future 

1929 examination. 

1930 

1931 Statistical analyses : Normally distributed data were analyzed using the Dunnett test, based on a 

1932 pooled variance estimate. The Steel test was applied if data were not normally distributed. The 

1933 Fisher Exact test was applied to frequency data. All tests were two-sided, with p < 0.05 accepted 

1934 as the lowest level of significance . No statistical analyses were performed on histopathological 

1935 data. The following were calculated for each group: percentage mating, fertility index, 

1936 conception rate, gestation index, and duration of gestation. Data were expressed as a mean +/-

1937 standard deviation of the number observed for each litter, and relative to a second parameter and 

1938 calculated on a total group basis. The following were calculated for each group: percentage of 

1939 live males at first litter check, percentage of live females at first litter check, percentage of 

1940 postnatal loss from Days 0-4 postpartum, and viability index. Data were expressed as a mean +/-

1941 standard deviation of the number observed for each litter, and relative to a second parameter and 

1942 calculated on a total group basis. 

1943 

1944 RESULTS 

1945 

1946 NOAEL (Parental FO) = 300 mg/kg bw/day 

1947 NOAEL (Offspring Fl) =1000 mg/kg bw/day 

1948 

1949 Remarks: Summary Table: 

1950 Parameter/Control/100 mg/kg/300 mg/kg/1000 mg/kg 

1951 Percentage/100/100/100/100 

1952 Mating fertility index/80/100/90/100 

1953 Gestation index/100/100/100/100 

1954 Mean Precoital time/2 .5/2 .9/1 .4/1 .7 

1955 Duration of gestation (days)/21 .1 +/- 0.4/21 .1 +/- 0.3/21 .0 +/- 0/21 .1 +/- 0.3 

1956 Dead pups at first litter check/0/0/0/0 

1957 Postnatal Loss(%)/0/2.6/2 .9/1 .0 

1958 Liters affected/0/3/2/1 

1959 Viability index/ 100/97.4/97.1/99 

1960 

1961 Parental animals: 1000 mg/kg bw/day: None of the animals died and functional observations 

1962 were not affected. One female showed hunched posture and piloerection from Day 20 post1963 coitum until Day 3 of lactation. Piloerection was noted in another female on Day 4 of lactation. 

1964 Slight salivation noted incidentally in both sexes was attributed to the test of the formulation. 

1965 Lower mean body weights were recorded for males and females at 1000 mg/kg during the 

1966 complete treatment period (not always statistically significant) . Body weight gains of males were 

1967 also lower than control throughout the treatment period. Food consumption before or after 

1968 allowance for body weight was reduced in males and females during the whole observation 

1969 period (not always statistically significant) . A slightly higher white blood cell count was noted in 
1970 males (not statistically significant) and females. Relative numbers of reticulocytes were 

1971 increased in males (3 .6 +/- 0.8% in treated vs. 2.3 +/- 0.1 % in control). Increased mean 

1972 corpuscular volume and decreased mean corpuscular hemoglobin concentration in females were 

1973 not considered to be related to treatment as they were either within the range of normal or 

1974 showed no dose-response relationship . Increased alanine aminotransferase was found in males 

1975 (59.2 +/- 20.4 U/1 in treated vs. 38.0 +/- 3.6 U/1 in control) and two females. One male had 

1976 increased activity of aspartate aminotransferase. Increased total bilirubin and decreased sodium 

1977 in males were not considered to be related to treatment to the lack of a dose-response 

1978 relationship . One male showed exophthalmus of the eye, which was considered to be caused by 

1979 blood sampling. Increases in organ/body weight ratios for the liver (2.57 +/- 0.14% in treated vs . 

198o 2 .33 +/- 0.12 % in control), testes (1 .16 +/- 0.15% in treated vs. 0.94 +/- 0.16% in control) and 

1981 epididymides (0.388 +/- 0.071% in treated vs. 0.301 +/- 0.050 in control) of males and liver 

1982 (3 .64 +/- 0.22% in treated vs. 3 .04 +/- 0.33% in control) spleen (0.287 +/- 0.023% in treated vs. 

1983 0.225 +/- 0.0303% in control), kidneys (0.83 +/- 0.05% in treated vs. 0.73 +/- 0.06% in control) 

1984 and adrenals (0.043 +/- 0.004% in treated vs. 0.034 +/- 0.004% in control) of females were 

1985 observed. There were no corresponding histological changes. An increase in the organ/body 

1986 weight ratio of the brain in males (0.66 +/- 0.05% in treated vs. 0.60 +/- 0.01% in control) was 

1987 considered to be due to the relatively low terminal body weight (300 +/- 14 g in treated vs. 336 

1988 +/- 19 g in control). Absolute brain weight (but not relative) was slightly decreased in females 

1989 (1 .77 +/- 0.05 g in treated vs. 1 .87 +/- 0.05 in control). Reproducitve .and breeding parameters 

1990 were not affected by treatment with up to 1000 mg/kg/day. 

1991 

1992 300 mg/kg bw/day: None of the animals died and functional observations were not affected. 

1993 Females had slightly lower mean absolute body weights on Day 8 of premating (189 +/- 6.6 g in 

1994 treated vs. 197 +/- 7.2 g in control). As this had recovered on Day 1 of mating (198 +/- 11 .8 g in 

1995 treated vs. 203 +/- 8.2 g in control), it was not considered to be toxicologically relevant. 

1996 Increased basophils in males, increased mean corpuscular volume and mean corpuscular 

1997 hemoglobin in females, decreased total bilirubin in males, and decreased albumin in females 

1998 were not considered to be related to treatment as they were either within the range of normal or 

1999 showed no dose-response relationship . No other abnormal findings were noted. 

2000 100 mg/kg bwlday: None of the animals died and functional observations were not affected. 

2001 Females had slightly lower mean absolute body weights on Day 8 of premating (189 +/- 6 .6 g in 

2002 treated vs. 197 +/- 7.2 g in control) . As this had recovered on Day 1 of mating (196 +/- 6.3 g in 

2003 treated vs. 203 +/- 8 .2 g in control), it was not considered to be toxicologically relevant . 

2004 Increased basophils in males and decreased total bilirubin in males were not considered to be 

2005 related to treatment as they were either within the range of normal or showed no dose-response 

2006 relationship . No other abnormal findings were noted. 

2007 

2008 Control: One female showed exophthalmus of the eye, which was considered to be caused by 

2009 blood sampling. 

2010 

2011 Offspring: Postnatal loss and viability index were similar for the control and treated groups. 

2012 Development of pups was not affected by treatment with up to 1000 mg/kg bw/day. Mean body 

2013 weights were similar for the control and treated groups (means ranged from 5 .6 - 5 .8 g and 5 .3 -

2014 5.6 g in males and females on Day 1, and 8.3 - 8.8 and 8 .0 - 8 .6 g in males and females on 

2015 Day 4, with none of the low values in the listed ranges being in high dose animals). None of the 
2016 clinical or macroscopic findings (scabs at the neck, dark red discoloration of tail apex, blue spot 

2017 on neck, wound at the head) were considered to be related to treatment as they were considered 

2018 to be within normal biological variation. 

2019 

2020 CONCLUSIONS 

2021 

2022 A limit dose of 1000 mg/kg/day was selected as the high dose level based on data obtained in a 

2023 preliminary range-finding study. The authors reported no-observable-effect-levels (NOAELs) of 

2024 300 mg/kg for general toxicity and 1000 mg/kg for reproductive and neurological effects. 

2025 

2026 RELIABILITY CODE 

2027 

2028 1- Valid Without Restrictions 

2029 Flag: Critical study for SIDS endpoint 

2030 

2031 REFERENCE 

2032 

2033 Beekhuijzen, M.E.W. 2008. A combined 28-day repeated dose toxicity study with the 

2034 reproduction/developmental toxicity screening test of Methane sulfinic acid, hydroxy-, 

2035 monosodium salt in rats by oral gavage. NOTOX B.V. Report No. 485218 (485615), dated 01-

2036 15-2008 (first draft of an unpublished study report) 

2037 

2038 Robust Summaries and Dossier for CAS No. 149-44-0, PCA Services, Inc. 

2039 

2040 

2041 

2042 

2043 

2044 

2045 

2046 

2047 

2048 

2049 

2050 

2051 

2052 

2053 

2054 

2055 

2056 

2057 

2058 

2059 

2060 

2061 
Y 

2062 DEVELOPMENTAL TOXICITY 

2063 

2064 Summary of endpoint: Adequate 

2065 

2066 Title: Repeated-dose/reproductive/developmental toxicity study 

2067 

2068 The study is : ® Adequate F~ Not Adequate 

2069 

2070 

2071 TEST SUBSTANCE 

2072 

2073 Identity: Methanesulfinic acid, hydroxyl-, monosodium salt (CAS No. 149-44-0) 

2074 

2075 Remarks: Analytical purity : 100.3% calculated as dihydrate. Batch Number 07041205 

2076 from Bruggemann. 

2077 

2078 METHOD 

2079 

2080 Method/guideline followed : OECD 422 

2081 

2082 Study duration: Males were exposed for 30 days (during 2 weeks prior to mating, during 

2083 mating, and to termination) . Females were exposed for 42-53 days (during 2 weeks prior to 

2084 mating, during mating and gestation, up to Day 4 of lactation) . 

2085 

2086 Test type: combined 28-day repeated dose toxicity study with the reproductive/developmental 

2087 toxicity screening test by oral gavage 

2088 

2089 GLP (Y/N): Yes 

2090 

2091 Year study performed: 2008 

2092 

2093 Species/strain: Wistar Rat 

2094 

2095 Sex: Both M/F 

2096 

2097 Number of animals per dose groups: Four groups of 10/sex 

2098 

2099 Route of administration: Oral gavage 

2100 

2101 Doses tested and frequency: 100, 300, 1000 mg/kg/day 

2102 

2103 Vehicle: Water 

2104 

2105 Post-observation period: Not stated 

2106 

2107 Statistical methods used: Normally distributed data were analyzed using the Dunnett test, based 
2108 on a pooled variance estimate . The Steel test was applied if data were not normally distributed. 

2109 The Fisher Exact test was applied to frequency data. All tests were two-sided, with p < 0.05 

2110 accepted as the lowest level of significance . No statistical analyses were performed on 

2111 histopathological data. 

2112 

2113 Remarks: 

2114 Frequency of Treatment: daily 

2115 

2116 Method/Guideline and Test Condition Remarks: Type: other: combined 28-day repeated dose 

2117 toxicity study with the reproductive/developmental toxicity screening test by oral gavage (OECD 

2118 422 and EPA, 40 CFR 799.9365) 

2119 

2120 Exposure period: Males were exposed for 30 days (during 2 weeks prior to mating, during 

2121 mating, and to termination). Females were Dosed for 42 - 53 days (during 2 weeks prior to 

2122 mating, during mating and gestation, up to Day 4 of lactation) 

2123 

2124 Methods pertaining to repeated dose and reproductive toxicity are presented in section 5.8 .1 . 

2125 Pups were killed by decapitation on Day 5 of lactation (or shortly thereafter). All offspring were 

2126 sexed and externally examined. Stomachs were examined for the presence of milk. Descriptions 

2127 of all external abnormalities were recorded. If possible, defects or causes of death were 

2128 evaluated. Any abnormal pup, organ or tissue was preserved for possible future examination. 

2129 Statistical analyses : The following were calculated for each group: percentage of live males at 

2130 first litter check, percentage of live females at first litter check, percentage of postnatal loss from 

2131 Days 0-4 post partum, and viability index. Data were expressed as a mean +/- standard deviation 

2132 of the number observed for each litter, and relative to a second parameter and calculated on a 

2133 total group basis . 

2134 

2135 RESULTS 

2136 

2137 NOAEL (Female, maternal) = 300 mg/kg bw/day 

2138 NOAEL (Fetal) =1000 mg/kg bw/day 

2139 

2140 Remarks: Remark: A limit dose of 1000 mg/kg/day was selected as the high dose level based on 

2141 data obtained in a preliminary range-finding study. The authors reported no-observable-effect2142 levels (NOAELs) of 300 mg/kg for general toxicity and 1000 mg/kg for reproductive and 

2143 neurological effects . 

2144 

2145 Results pertaining to repeated dose and reproductive toxicity are presented in section 5.8 .1 . 

2146 

2147 

2148 

2149 

2150 

2151 

2152 

2153 

Offspring: Postnatal loss and viability index were similar for the control and treated groups . 

Development of pups was not affected by treatment with up to 1000 mg/kg bw/day. Mean body 

weights were similar for the control and treated groups (means ranged from 5 .6 - 5.8 g and 5.3 -

5.6 g in males and females on Day 1, and 8.3 - 8.8 and 8.0 - 8.6 g in males and females on Day 4, 

with none of the low values in the listed ranges being in high dose animals) . None of the clinical 

or macroscopic findings (scabs at the neck, dark red discoloration of tail apex, blue spot on neck, 

wound at the head) were considered to be related to treatment as they were considered to be 
p. 60 

. 

.. 2154 within normal biological variation. 

2155 

2156 Parameter/control/ 100 mg/kg / 300 mg/kg / 1000 mg/kg 

2157 Dead pups at first litter check/0/0/0/0 

2158 Postnatal Loss (%)/0/2.6/2 .9/1 .0 

2159 Litters Affected/0/3/2/1 

2160 Viability Index/100/97 .4/97.1/99.0 

2161 

2162 CONCLUSIONS 

2163 

2164 Result: not a developmental toxicant 

2165 

2166 RELIABILITY CODE 

2167 

2168 1 - Valid Without Restrictions 

2169 Flag: critical study for SIDS endpoint 

2170 

2171 REFERENCE 

2172 

2173 Beekhuijzen, M.E.W. 2008 . A combined 28-day repeated dose toxicity study with the 

2174 reproduction/developmental toxicity screening test of Methane sulfinic acid, hydroxy-, 

2175 monosodium salt in rats by oral gavage. NOTOX B.V. Report No. 485218 (485615), dated O1-

2176 15-2008 (first draft of an unpublished study report) 

2177 

2178 Robust Summaries and Dossier for CAS No . 149-44-0, PCA Services, Inc. 

2179 

2180 

2181 

2182 

2183 

2184 

2185 . 

2186 

2187 

2188 

2189 

2190 

2191 

2192 

2193 

2194 

2195 

2196 

2197 

2198 

2199 
2200 

2201 

2202 

2203 

Summary of Endpoints 

Endpoint ~ Study Information 

Provided 

Acceptable Data Gap 

Aquatic Effects 

Acute Toxicity to Fish Yes Yes No 

Acute Toxicity to Daphnia Yes Yes No 

Toxicity to A uatic Plants ~ Yes ~ Yes ~ No 

- 

Acute Toxicity Yes No Yes 

Repeated-Dose Toxicity Yes Yes No 

Genetic Mutation Yes No Yes 

Chromosomal Aberrations Yes Yes No 

Toxicity to Reproduction Yes Yes No 

Developmental Toxicity ~ Yes ~ Yes ~ No 

1180 ON SNIdIN00