### SRC TR

- 09

- 219

HPV4 CHEMICAL DATA ADEQUA CY REVIEW AND ROBUST SUMMARY ADEQUACY

REQUIREMENTS

Quaternary ammonium compounds, benzyl(hydrogenated tallow alkyl)dimethyl, chlorides

Prepared by:

Emily Cohen, Christopher Janson, 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 FA411.4.409.009

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

July 16, 2009

### TOXICITY TO AQUATIC VERTEBRATES

Summary of endpoint: Adequate

Title: Daily Static

- Renewal Acute 96

- Hour Toxicity Test of Alkyl Dimethyl B enzyl Ammonium

Chloride (ADBAC) to Bluegill Sunfish

The study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: ADBAC, CAS No. 68424

- 85

- 1

Remarks: 30% ai

METHOD

Method/guideline followed: OPPTS 850.1075

GLP: Not stated

Year study performed: Not stated

Test type : Static

- renewal

Species/strain/supplier: Bluegill sunfish ( Lepomis macrochirus )

Analytical monitoring: Not stated

Exposure period: 96 hours

Number of animals/sex/dose: Not stated

Statistica l methods used: Not stated

Method remarks: Not stated

RESULTS

Nominal concentrations: Not stated

Measured concentrations: Not stated

Element value: LC50 = 0.515 mg/L; NOAEC = 0.456 mg/L (a.i.)

Remarks: Highly toxic

CONCLUSIONSCategorized as h ighly toxic to fish

RELIABILITY CODE

2

REFERENCES

MRID 419472

- 01. Pate, H.O. and D.O. McIntyre. 1991. Daily Static

- Renewal Acute 96

- Hour Toxicity

Test of Alkyl Dimethyl Benzyl Ammonium Chloride (ADBAC) to Bluegill Sunfish. Study No.

SC890050. Unpu blished data. Conducted by Batelle Columbus Division for ADBAC Quat Joint

Venture/Chemical Specialties Manufacturers Association.

(EPA

- HQ

- OPP

- 2006

- 0339

- 0021[1].pdf)Title: Static

- Renewal Acute Toxicity of ADBAC to Fathead Minnow ( Pimephales promela s )

The study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: ADBAC, CAS No. 68424

- 85

- 1

Remarks: 80% ai

METHOD

Method/guideline followed: Not stated

GLP: Not stated

Year study performed: Not stated

Test type: Static

- renewal

Species/strain/supplier: Fathead minnow ( Pimephales promelas )

Analytical monitoring: Not stated

Exposure period: 96 hours

Number of animals/sex/dose: Not stated

Statistical methods used: Not stated

Method remarks: Not state d

RESULTS

Nominal concentrations: Not stated

Measured concentrations: Not stated

Element value: LC50 = 0.28 mg/L; NOAEC = Not Determined (a.i.)

Remarks: Highly toxic

CONCLUSIONS

Categorized as highly toxic to fish

RELIABILITY CODE

2REFERENCES

MRID 437401

- 03. Sword, M.C. and L. Stuerman. 1993. Static

- Renewal Acute Toxicity of ADBAC to

Fathead Minnow ( Pimephales promelas ). Study No. 41237. Unpublished data. Conducted by ABC

Laboratories Inc. for ADBAC Quat Joint Venture/Chemical Specialti es Manufacturers Association.

(EPA

- HQ

- OPP

- 2006

- 0339

- 0021[1].pdf)Title: Static

- Renewal Acute Toxicity of ADBAC to Fathead Minnow ( Pimephales promelas ) in Dilution

Water Amended with 20 mg/L Humic Acid

The study is: Adequate Not Adequate

This study was considered inadequate due to deviations from the guideline, including the addition of 20

mg/L humic acid to dilution water.

TEST SUBSTANCE

Identity: ADBAC, CAS No. 68424

- 85

- 1

Remarks: 80% ai

METHOD

Method/guide line followed: Not stated

GLP: Not stated

Year study performed: Not stated

Test type: Static

- renewal

Species/strain/supplier: Fathead minnow ( Pimephales promelas )

Analytical monitoring: Not stated

Exposure period: 96 hours

Number of animals/sex/do se: Not stated

Statistical methods used: Not stated

Method remarks: Deviations from guideline include additions of 20 mg/L humic acid to dilution water

RESULTS

Nominal concentrations: Not stated

Measured concentrations: Not stated

Element value: LC50 = 1.4 mg/L; NOAEC = 0.99 mg/L (a.i.)

Remarks: Moderately toxic

CONCLUSIONS

Categorized as moderately toxic to fish

RELIABILITY CODE3

REFERENCES

MRID 437401

- 01 . Sword, M.C. and L. Stuerman. 1993. Static

- Renewal Acute Toxicity of ADBAC to

Fathead Minnow ( Pimephales promelas ) in Dilution Water Amended with 20 mg/L Humic Acid. Study

No. 41235. Unpublished data. Conducted by ABC Laboratories, Inc. for ADBAC Quat Joint

Venture/Chemical Specialties Manufacturers Association.

(EPA

- HQ

- OPP

- 2006

- 0339

- 0021[1].pdf)Title: Static

- Renewal Acute Toxicity of ADBAC to Fathead Minnow ( Pimephales promelas ) in Dilution

Water Amended with 10 mg/L Humic Acid

The study is: Adequate Not Adequate

This study was conside red inadequate due to deviations from the gui deline, including the addition of 1 0

mg/L humic acid to dilution water.

TEST SUBSTANCE

Identity: ADBAC, CAS No. 68424

- 85

- 1

Remarks: 80% ai

METHOD

Method/guideline followed: Not stated

GLP: Not stated

Y ear study performed: Not stated

Test type: Static

- renewal

Species/strain/supplier: Fathead minnow ( Pimephales promelas )

Analytical monitoring: Not stated

Exposure period: 96 hours

Number of animals/sex/dose: Not stated

Statistical methods used: Not stated

Method remarks: Guideline deviations include 10 mg/L humic acid added to dilution water

RESULTS

Nominal concentrations: Not stated

Measured concentrations: Not stated

Element value: LC50 = 0.77 mg/L; NOAEC = 0.53 mg/L (a.i.)

Remarks: H ighly toxic

CONCLUSIONS

Categorized as highly toxic to fish

RELIABILITY CODE3

REFERENCES

MRID 437401

- 02. Sword, M.C. and L. Stuerman. 1993. Static

- Renewal Acute Toxicity of ADBAC to

Fathead Minnow ( Pimephales promelas ) in Dilution Water Amended with 10 mg/L Humic Acid. Study

No. 41236. Unpublished data. Conducted by ABC Laboratories, Inc. for ADBAC Quat Joint

Venture/Chemical Specialties Manufacturers Association.

(EPA

- HQ

- OPP

- 2006

- 0339

- 0021[1].pdf)Title: Environmental Assessment of an Alky l Dimethyl Benzyl Ammonium Chloride (ADBAC) Based

Molluscicide Using Laboratory Tests

The study is: Adequate Not Adequate

This study was considered inadequate due to missing critical study information.

TEST SUBSTANCE

Identity: ADBAC, CAS No. 68424

- 85

- 1

Remarks: 50% ai

METHOD

Method/guideline followed: Not stated

GLP: Not stated

Year study performed: Not stated

Test type: Flow

- through

Species/strain/supplier: Fathead minnow ( Pimephales promelas )

Analytic al monitoring: Not stated

Exposure period: 96 hours

Number of animals/sex/dose: Not stated

Statistical methods used: Not stated

Method remarks: Not stated

RESULTS

Nominal concentrations: Not stated

Measured concentrations: Not stated

Element v alue: LC50 = 0.36 mg/L (a.i.)

Remarks: Highly toxic

CONCLUSIONS

Categorized as highly toxic to fish

RELIABILITY CODE3

– Missing data

REFERENCES

Dobbs, M.G., D.S. Cherry, J.C. Scott, and J.C. Petrille. 1995. Environmental Assessment of an Alkyl

Dimethyl Benzyl Ammonium Chloride (ADBAC) Based Molluscicide Using Laboratory Tests.

Published data. Proceedings of the Fifth International Zebra Mussel and Other Aquatic Nuisance

Organisms Conference, Toronto, Canada.

(EPA

- HQ

- OPP

- 2006

- 0339

- 0021[1])T itle: Daily Static

- Renewal Acute 96

- Hour Toxicity Test of Alkyl Dimethyl Benzyl Ammonium

Chloride (ADBAC) to Rainbow Trout

The study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: ADBAC, CAS No. 68424

- 85

- 1

Re marks: 30% ai

METHOD

Method/guideline followed: Not stated

GLP: Not stated

Year study performed: Not stated

Test type: Static

- renewal

Species/strain/supplier: Rainbow trout ( Oncorhynchus mykiss )

Analytical monitoring: Not stated

Exposure perio d: 96 hours

Number of animals/sex/dose: Not stated

Statistical methods used: Not stated

Method remarks: Not stated

RESULTS

Nominal concentrations: Not stated

Measured concentrations: Not stated

Element value: LC50 = 0.923 mg/L; NOAEC = 0.619 mg /L (a.i.)

Remarks: Highly toxic

CONCLUSIONS

Categorized as highly toxic to fish

RELIABILITY CODE

2REFERENCES

MRID 419472

- 02. Pate, H.O. and D.O. McIntyre. 1991. Daily Static

- Renewal Acute 96

- Hour Toxicity

Test of Alkyl Dimethyl Benzyl Ammonium Chloride (ADBAC) to Rainbow Trout. Study No.

SC890051. Unpublished data. Conducted by Batelle Columbus Division for ADBAC Quat Joint

Venture/Chemical Specialties Manufacturers Association.

(EPA

- HQ

- OPP

- 2006

- 0339

- 0021[1])Title: Environmental Asse ssment of an Alkyl Dimethyl Benzyl Ammonium Chloride (ADBAC) Based

Molluscicide Using Laboratory Tests

The study is: Adequate Not Adequate

This study was considered inadequate due to missing critical study information

.

TEST SUBSTANCE

Identity: ADBAC, CAS No. 68424

- 85

- 1

Remarks: 50% ai

METHOD

Method/guideline followed: Not stated

GLP: Not stated

Year study performed: Not stated

Test type: Flow

- through

Species/strain/supplier: Rainbow trout ( Oncorhynchus my kiss )

Analytical monitoring: Not stated

Exposure period: 96 hours

Number of animals/sex/dose: Not stated

Statistical methods used: Not stated

Method remarks: Not stated

RESULTS

Nominal concentrations: Not stated

Measured concentrations: Not stated

Element value: LC50 = 1.01 mg/L (a.i.)

Remarks: Highly/moderately toxic

CONCLUSIONS

Categorized as highly/moderately toxic to fish

RELIABILITY CODE3

– Data missing

REFERENCES

Dobbs, M.G., D.S. Cherry, J.C. Scott, and J.C. Petrille. 1995. Environmental Assessment of an Alkyl

Dimethyl Benzyl Ammonium Chloride (ADBAC) Based Molluscicide Using Laboratory Tests.

Published data. Proceedings of the Fifth International Zebra Mussel and Other Aquatic Nuisance

Organisms Conference, Toronto, Cana da.

(EPA

- HQ

- OPP

- 2006

- 0339

- 0021[1])Title: A 96

- Hour Static Renewal Acute Toxicity Test with Alkyl Dimethyl Benzyl Ammonium Chloride

(ADBAC) in the Sheepshead Minno ( Cyprinodon variegatus )

The study is: Adequate N ot Adequate

TEST SUBSTANCE

Identity: ADBAC, CAS No. 68424

- 85

- 1

Remarks: 80% ai

METHOD

Method/guideline followed: OPPTS 850.10 7 5

GLP: Not stated

Year study performed: Not stated

Test type: Static

- renewal

Species/strain/supplier: Sheepshead min now ( Cyprinodon variegatus )

Analytical monitoring: Not stated

Exposure period: 96 hours

Number of animals/sex/dose: Not stated

Statistical methods used: Not stated

Method remarks: Not stated

RESULTS

Nominal concentrations: Not stated

Measured concentrations: Not stated

Element value: LC50 = 0.86 mg/L (a.i.); NOAEC = 0.68 mg/L (a.i.)

Remarks: Highly toxic

CONCLUSIONS

Categorized as highly toxic to fish

RELIABILITY CODE

2REFERENCES

MRID 424795

- 02 . Sved, D.W., J.P. Swigert, and G.J. S mith. 1992. A 96

- Hour Static Renewal Acute

Toxicity Test with Alkyl Dimethyl Benzyl Ammonium Chloride (ADBAC) in the Sheepshead Minno

( Cyprinodon variegatus ). Project No. 350A

- 102. Unpublished data. Conducted by Wildlife

International Ltd. for ADBAC Q uat Joint Venture/Chemical Specialties Manufacturers Association.

(EPA

- HQ

- OPP

- 2006

- 0339

- 0021[1])Title : Environmental Assessment of an Alkyl Dimethyl Benzyl Ammonium Chloride (ADBAC) Based

Molluscicide Using Laboratory Tests

The study is: Adequate Not Adequate

This study was considered inadequate due to missing critical study information as well as testing of a

non

- standard species.

TEST SUBSTANCE

Identity: ADBAC, CAS No. 68424

- 85

- 1

Remarks: 50% ai

METHOD

M ethod/guideline followed: Not stated

GLP: Not stated

Year study performed: Not stated

Test type: Flow

- through

Species/strain/supplier: Inland silverside (Menidia beryllina )

Analytical monitoring: Not stated

Exposure period: 96 hours

Number of a nimals/sex/dose: Not stated

Statistical methods used: Not stated

Method remarks: Not stated

RESULTS

Nominal concentrations: Not stated

Measured concentrations: Not stated

Element value: LC50 = 0.31 mg/L (a.i.)

Remarks: Highly toxic

CONCLUSION S

Categorized as highly toxic to fish

RELIABILITY CODE3

- Data missing; Non

- guideline species

REFERENCES

Dobbs, M.G., D.S. Cherry, J.C. Scott, and J.C. Petrille. 1995. Environmental Assessment of an Alkyl

Dimethyl Benzyl Ammonium Chloride (ADBAC) B ased Molluscicide Using Laboratory Tests.

Published data. Proceedings of the Fifth International Zebra Mussel and Other Aquatic Nuisance

Organisms Conference, Toronto, Canada.

(EPA

- HQ

- OPP

- 2006

- 0339

- 0021[1])

### TOXICITY TO AQUATIC INVERTEBRATES

Summa ry of endpoint: Adequate

Title: Daily Static

- Renewal Acute 48

- Hour Toxicity Test of Alkyl Dimethyl Ammonium Chloride

(ADBAC) to Daphnia magna

.

The study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: ADBAC , CAS No. 68424

- 85

- 1

Remarks: 30% ai

METHOD

Method/guideline followed: OPPTS 850.1010

GLP: Not stated

Year study performed: Not stated

Test type: Static

- renewal

Species/strain/supplier: Water flea ( Daphnia magna )

Analytical monitoring: Not stated

Exposure period: 48 hours

Number of animals/sex/dose: Not stated

Statistical methods used: Not stated

Method remarks: Not stated

RESULTS

Nominal concentrations: Not stated

Measured concentrations: Not stated

Element value: EC50 = 0.0059 mg/L; NOAEC = Not Determined (a.i.)

Remarks: Very highly toxic

CONCLUSIONSCategorized as very highly toxic to aquatic invertebrates.

RELIABILITY CODE

2

REFERENCES

MRID 419472

- 03 . Pate, H.O. and D.O. McIntyre. 1991. Daily Static

- Renewal Acute 48

- Ho ur Toxicity

Test of Alkyl Dimethyl Ammonium Chloride (ADBAC) to Daphnia magna . Study No. SC 89005 2

. Unpublished data. Conducted by Batelle Columbus Division for ADBAC Quat Joint Venture/Chemical

Specialties Manufacturers Association.

(EPA

- HQ

- OPP

- 2006

- 0 339

- 0021[1])Title: Environmental Assessment of an Alkyl Dimethyl Benzyl Ammonium Chloride (ADBAC) Based

Molluscicide Using Laboratory Tests

The study is: Adequate Not Adequate

This study was considered inadequate due to missing critical study information.

TEST SUBSTANCE

Identity: ADBAC, CAS No. 68424

- 85

- 1

Remarks: 50% ai

METHOD

Method/guideline followed: Not stated

GLP: Not stated

Year study performed: Not stated

Test type: Flow

- through

Species/strain/s upplier: Water flea ( Daphnia magna )

Analytical monitoring: Not stated

Exposure period: 48 hours

Number of animals/sex/dose: Not stated

Statistical methods used: Not stated

Method remarks: Not stated

RESULTS

Nominal concentrations: Not stated

Mea sured concentrations: Not stated

Element value: LC50 = 0.02 mg/L (a.i.)

Remarks: Very highly toxic

CONCLUSIONS

Categorized as very highly toxic to aquatic invertebrates.

RELIABILITY CODE3

- Data Missing

REFERENCES

Dobbs, M.G., D.S. Cherry, J.C. Scott, and J.C. Petrille. 1995. Environmental Assessment of an Alkyl

Dimethyl Benzyl Ammonium Chloride (ADBAC) Based Molluscicide Using Laboratory Tests.

Published data. Proceedings of the Fifth International Zebra Mussel and Other Aquatic Nuisance

Or ganisms Conference, Toronto, Canada.

(EPA

- HQ

- OPP

- 2006

- 0339

- 0021[1])Title: A 96

- Hour Static

- Renewal Acute Toxicity Test with Alkyl Dimethyl Benzyl Ammonium Chloride

(ADBAC) in the Saltwater Mysid (Mysidopsis bahia)

The study is: Ade quate Not Adequate

TEST SUBSTANCE

Identity: ADBAC, CAS No. 68424

- 85

- 1

Remarks: 80% ai

METHOD

Method/guideline followed: OPPTS 850.10 3 5

GLP: Not stated

Year study performed: Not stated

Test type: Static

- renewal

Species/strain /supplier: Mysid shrimp ( Mysidopsis bahia )

Analytical monitoring: Not stated

Exposure period: 96 hours

Number of animals/sex/dose: Not stated

Statistical methods used: Not stated

Method remarks: Not stated

RESULTS

Nominal concentrations: Not stat ed

Measured concentrations: Not stated

Element value: LC50 = 0.092 mg/L (a.i.); NOAEC = 0.047 mg/L (a.i.)

Remarks: Very highly toxic

CONCLUSIONS

Categorized as very highly toxic to aquatic invertebrates.

RELIABILITY CODE

2REFERENCES

MRID 4247 95

- 01 . Sved, D.W., J.P. Swigert, and G.J. Smith. 1992. A 96

- Hour Static

- Renewal Acute

Toxicity Test with Alkyl Dimethyl Benzyl Ammonium Chloride (ADBAC) in the Saltwater Mysid

(Mysidopsis bahia). Project No. 350A

- 101A. Unpublished data. Conducted by Wildlife International

Ltd. for ADBAC Quat Joint Venture/Chemical Specialties Manufacturers Association.

(EPA

- HQ

- OPP

- 2006

- 0339

- 0021[1])Title: Environmental Assessment of an Alkyl Dimethyl Benzyl Ammonium Chloride (ADBAC) Based

Molluscicide Using Labo ratory Tests

The study is: Adequate Not Adequate

This study was considered inadequate due to missing critical study information.

TEST SUBSTANCE

Identity: ADBAC, CAS No. 68424

- 85

- 1

Remarks: 50% ai

METHOD

Method/ guideline followed: Not stated

GLP: Not stated

Year study performed: Not stated

Test type: Flow

- through

Species/strain/supplier: Mysid shrimp ( Mysidopsis bahia )

Analytical monitoring: Not stated

Exposure period: 96 hours

Number of animals/sex/dose: Not stated

Statistical methods used: Not stated

Method remarks: Guideline deviations included testing conducted at higher temperature.

RESULTS

Nominal concentrations: Not stated

Measured concentrations: Not stated

Element value: LC50 = 0.08 mg/L (a.i.)

Remarks: Very highly toxic

CONCLUSIONS

Categorized as very highly toxic to aquatic invertebrates.

RELIABILITY CODE3

- Data missing

REFERENCES

Dobbs, M.G., D.S. Cherry, J.C. Scott, and J.C. Petrille. 1995. Environmental Assessment of an Alkyl

Dimethyl Benzyl Ammonium Chloride (ADBAC) Based Molluscicide Using Laboratory Tests.

Published data. Proceedings of the Fifth International Zebra Mussel and Other Aquatic Nuisance

Organisms Conference, Toronto, Canada.

(EPA

- HQ

- OPP

- 2006

- 0339

- 0021[1 ])Title: A 48

- Hour Static Acute Toxicity Test with Alkyl Dimethyl Benzyl Ammonium Chloride

(ADBAC) in Embryo Larvae of the Eastern Oyster ( Crassostrea virginica )

The study is: Adequate Not Adequate

This study w as considered inadequate due to the presence of high mortality in control groups.

TEST SUBSTANCE

Identity: ADBAC, CAS No. 68424

- 85

- 1

Remarks: 80% ai

METHOD

Method/guideline followed: Not stated

GLP: Not stated

Year study performed: Not stated

Tes t type: Static

Species/strain/supplier: Eastern oyster ( Crassostrea virginica ) (embryo larvae)

Analytical monitoring: Not stated

Exposure period: 48 hours

Number of animals/sex/dose: Not stated

Statistical methods used: Not stated

Method remarks: None stated

RESULTS

Nominal concentrations: Not stated

Measured concentrations: Not stated

Element value: LC50 = 0.055 mg/L (a.i.)

Remarks: Very highly toxic

CONCLUSIONS

Categorized as very highly toxic to aquatic invertebrates.

RELIABILITY CO DE3

- High control mortality

REFERENCES

MRID 424795

- 03. Sved, D.W., J.P. Swigert, and G.J. Smith. 1992. A 48

- Hour Static Acute Toxicity

Test with Alkyl Dimethyl Benzyl Ammonium Chloride (ADBAC) in Embryo Larvae of the Eastern

Oyster ( Crassostrea virg inica ). Unpublished data. Conducted by Wildlife International Ltd. for ADBAC

Quat Joint Venture/Chemical Specialties Manufacturers Assocation.

(EPA

- HQ

- OPP

- 2006

- 0339

- 0021[1])

### TOXICITY TO AQUATIC PLANTS

Summary of endpoint: Adequate

Title: Toxic Ef fects of Linear Alkylbenzene Sulfonate, Quaternary Alkylammonium Chloride and their

Complexes on Dunaliella sp. and Chlorella pyrenoidosa

The study is: Adequate Not Adequate

This study was considered inadequate due to testing of a non

- standard species.

TEST SUBSTANCE

Identity: Alkyldimethylbenzylammoniumchloride (R = C12

- 14); abbreviated as BM , CAS No. 68424

-

85

- 1

Remarks: Purity > 97.5%

METHOD

Method/guideline followed: Not stated

Test type: Not stated

GLP: N ot stated

Year: circa 1995

Species/strain/supplier: Dunaliella sp.

Element Basis: Not stated

Exposure Period: 24 hours

Analytical Monitoring: Not stated

Statistical Methods: Not stated

Method remarks:

The test chemical was dissolved with wat er to make a final concentration between 2 and 0.1 mM.

Dunaliella sp. isolated from an Antarctic high salt lake by Watanuki et al. was used to test the biological

effects of surfactacts by determining glycerol content of the cells. The precultures of tes t algae were

incubtated at 20 +/

- 1 C under continuous lightening at 140 μ E/m 2 /s and sterile aeration through 0.2

- μ m

filter for approximately 7 days. Modified Johnson’s medium was used in this study. The algal culture

was taken from a preculture in the e xponential growth phase. The initial cell density was about 6 x 10 6

cells/ml, as counted by Microcellcounter F

- 800. The 200

- ml algal culture and the stepwise graded

concentration of BM were added to 300

- ml culture bottles with two replicates of each conc entration.

These cultures were incubated without shaking and aeration of 24 h at 20 +/

- 1 C under continuouslightening at 140 μ E/m 2 /s using a controlled environmental incubator shaker. The number of cells in the

control was almost unchanged from the ini tial density during the 24

- h incubation. The coefficient of the

control cells was 15.1%

The glycerol contents at each concentration level of the test chemicals were measured by H

- NMR after 24

h. The dose

- response curves were drawn by plotting the glycer ol contents, which were shown with the

percent of untreated control without surfactant in the ordinate and the concentration of surfactant in the

abscissa. The 24

- h EC50 and 95% confidence level were calculated by Litchfield and Wilcoxon’s

method.

RESULT S

Nominal concentrations: Not stated

Measured concentrations: Not stated

Element value: 24

- hour EC 50 = 1.8 mg/L (effect on glycerol content)

Remarks: None

CONCLUSIONS

RELIABILITY CODE

3

REFERENCE

Utsunomiya, A., T. Watanuki, K. Matsushita, and I. Tomita , “ Toxic Effects of Linear Alkylbenzene

Sulfonate, Quaternary Alkylammonium Chloride and Their Complexes on Dunaliella sp. and Chlorella

pyrenoidosa ”. Environ.Toxicol.Chem. 16(6):1247

- 1254 ; 1997Title: Toxic Effects of Linear Alkylbenzene S ulfonate, Quaternary Alkylammonium Chloride and their

Complexes on Dunaliella sp. and Chlorella pyrenoidosa

The study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: Alkyldimethylbenzylammoniumchloride (R = C12

- 14); abbreviated as BM , CAS No. 68424

-

85

- 1

Remarks: Purity > 97.5%

METHOD

Method/guideline followed: Not stated

Test type: Not stated

GLP: Not stated

Year: circa 1995

Species/strain/supplier: Chlorella pyrenoidosa

Element Basis: area under the curve

Exposure Period: 96 hours

Analytical Monitoring: Not stated

Statistical Methods: Not stated

Method remarks: Chlorella pyrenoidosa was supplied by the Institute of Molecular and Cellular

Bioscience. The precultures of test alga were incubate d at 25 +/

- 1 C under continuous lightening at 140

μ E/m 2 /s and sterile aeration through a 0.2

- mm filter for approximately 7 days. A modified Bristol

medium was used.

The concentration ranges in which inhibitory effects were likely to occur were determine d from range

finding tests. The 400

- mL culture medium and the algal inoculums taken from a preculture in the

exponential growth phase were added to 500

- mL sterile culture bottles. The initial algal density was

approximately 1 x 10 4 cells/mL. The stepwi se graded concentrations of BM, with more than four

concentrations with two replicates each , were added to each test solution. The test solutions were

incubated at 24+/

- 2 C under continuous lightening at 140 μ E/m 2 /s with slow continuous shaking (100

rpm ) for 96 h using controlled environment incubator shaker. A 100

- to 10

- mL portion of each test

solution was centrifuged at 1,900 g for 15 min at 5 C. Chlorophyll

- a concentration, an indicator of cell

growth, was measured every 24 h from the start of this test to 96 h by Parsons and Strickland’s method.

The growth curves for each concentration of test substance and for the control were obtained by plotting

the chlorophyll

- a concentration in the ordinate and the incubation time in the abscissa. Following OECDtest guidelines, the areas under the growth curves were calculated every 24 h until 96 h, and the

percentage of growth inhibition was calculated from the ratio of the difference between the area under the

growth curve at each substance concentration. The 96

- h EC50 and 95% confidence level were calculated

by Litchfield and Wilcoxon’s method.

RESULTS

Nominal concentrations: Not stated

Measured concentrations: Not stated

Element value: EC50 = 0.67 mg/L

Remarks: None

CONCLUSIONS

RELIABILITY COD E

2

REFERENCE

Utsunomiya, A., T. Watanuki, K. Matsushita, and I. Tomita

Toxic Effects of Linear Alkylbenzene Sulfonate, Quaternary Alkylammonium Chloride and Their

Complexes on Dunaliella sp. and Chlorella pyrenoidosa

Environ.Toxicol.Chem. 16(6):1247

- 12 54

1997Title: Interpretation of Ultrastructral Changes in Cladophora glomerata Resulting from Hyamine

Toxicity

The study is: Adequate Not Adequate

This is not a guideline study and provides no LD50.

TEST SUBSTA NCE

Identity: Hyamine 3500, synonym for quaternary ammonium compounds, benzyl

- C12

- 16

- alkyldimethyl, chlorides , CAS No. 68424

- 85

- 1

Remarks: 80% concentrate in ethanol

METHOD

Method/guideline followed: Not stated

Test type: Not stated

GLP: Not stat ed

Year: circa 1992

Species/strain/supplier: Cladophora glomerata

Element Basis: Not stated

Exposure Period: 4 days

Analytical Monitoring: Not stated

Statistical Methods: Not stated

Method remarks: The test filamentous species, Cladophora glom erata (L.) Kutz. (UTTC #13) was

obtained from the University of Toronto Culture Collection (UTCC). The alga was originally isolated

from the Thames River, Ontario. The algal culture was maintained in Bold’s Basal Medium (BBM)

(Hutchinson and Stokes 1975 ) supplemented with a vitamin mix (B 12 , biotin and thiamine), and 5 mg/L Si

as NaSiO 3 x 9H 2 O

Chlorella fusca Shihira et Krauss, a unicellular alga, obtained from the laboratory at the Canada Centre

for Inland Waters, Burlington, Ontario, was the other spe cies used in the tests.

Hyamine 3500, was purchased from the Rohm and Haas Company. This algicide is an 80% concentrate

in ethanol, containing the active ingredient n

- alkyl (80% tetradecyl, 40% dodecyl and 10% hexadecyl)

dimethyl benzyl ammonium chloride . The molecular weight of the chemical is 359.6. Stable and

nonvolatile, it has been recommended by the company (Rohm and Haas 1978) as a disinfectant in

swimming pools at low concentrations.

Algal assays were conducted in duplicate in 250

- mL Erlenmeyer flasks containing 100 mL BBMsolution. The toxicity range of Hyamine for Cladophora was tested at the beginning of the study. In the

final assays, the Hyamine concentrations were set at 0.2, 0.3, 0.4, 0.5, 0.8 and 1.0 mg/L. Zero Hyamine

concentration w as used as a control. About 0.5 g (wet weight) of Cladophora filament was used as the

inoculums. The cultures were incubated for 4 days at 20 C in an environmental chamber where the light

and dark cycle was set at 12:12 and the light intensity was 60 μ Ei n/m 2 /sec.

On day 4, a small amount of plant material was removed from each concentration for morphological

examination using a light microscope. One set of samples was put aside for electron microscopic

preparation; the other set was incubated for anot her 3 weeks for further morphological evaluations. These

tests were repeated at the end of the experiment.

For electron microscope studies, Cladophora filaments were fixed in 4% (cacodylate

- buffered)

glutaraldehyde for 6 hr and then postfixed in 2% (ca codylate

- buffered) osium tetroxide for 1 hr. The

dehydration ethanol series was 30%, 50%, 70%, 90%, 95% and 100%.

For infiltration, the Spurr’s medium (Hayat 1989) to ethanol ratios were 1:3, 1:2, and 2:3 v/v. The

Cladophora filaments were then embedded in 100% Spurr’s medium in an aluminum dish. The samples

were polymerized at 70 C overnight.

A dissecting microscope was used to locate the apical cells on the plastic. After they were marked, small

columns of specimens were cut out with a jeweller’s s aw. Thin sections were prepared with a glass knife

on a microtome. These sections were then stained with uranyl acetate and lead citrate. Observations

were made using a Siemens electron microscope.

To assess the effect of Hyamine on Chlorella cells, a conventional method based on growth

measurements was used (Miller et al. 1978). A Chlorella pre

- culture was prepared. The Hyamine

concentrations used were the same as those employed in the Cladophora assays. On day 7, when the pre

culture reached its lo garithmic growth phase, the Chlorella cells were ready for inoculation. The initial

cell concentration in the test solution was 40 x10 3 cells/L. After inoculation, the culture flasks were

mounted on a platform shaker and incubated for 14 days in a growth chamber. Cells numbers were

enumerated at 4 day intervals and at the end of the experiment when the algal biomass reached its growth

plateau.

RESULTS

Nominal concentrations: 0.2, 0.3, 0.4, 0.5, 0.8 and 1.0 mg/L

Remarks:

Hyamine conc (mg/L) Chlorel la fusca avg. yield (10 5 cells/mL) after 14

days

control 14.5

0.15 13.6

0.20 15.3

0.30 13.9

0.40 no data

0.50 14.6

0.60 10.8

0.80 no growth1.00 no growth

CONCLUSIONS

RELIABILITY CODE

3

REFERENCE

Wong, S.L., and J.F. Wainwright . “ Interpreta tion of Ultrastructural Changes in Cladophora glomerata

Resulting from Hyamine Toxicity ”. Bull.Environ.Contam.Toxicol. 52(3):325

- 332 ; 1994Title: Environmental Assessment of an Alkyl Dimethyl Benzyl Ammonium Chloride (ADBAC) Based

Molluscicide Using L aboratory Tests.

The study is: Adequate Not Adequate

This study was considered inadequate due to deviations from accepting testing guidelines.

TEST SUBSTANCE

Identity: ADBAC, CAS No. 68424

- 85

- 1

Remarks: 50% (2.5 mg/L) plus Bentonite Clay at concentrations of 0:0, 2.5:25, 2.5:37.5, 2.5:50, and

2.5:75 mg/L

METHOD

Method/guideline followed: Not stated

Test type: Static

- renewal

GLP: Not stated

Year: Not stated

Species/strain/supplier: Green algae ( Selenastru m capricornutum )

Element Basis: Not stated

Exposure Period: 96 hours

Analytical Monitoring: Not stated

Statistical Methods: Not stated

Method remarks: None

RESULTS

Nominal concentrations: Not stated

Measured concentrations: Not stated

Eleme nt value: LOAEC = 2.5:25 for growth; NOAEC = 2.5:37.5 for growth

Remarks:

Supplemental study

Open literature

Assess detoxification of a.i.CONCLUSIONS

RELIABILITY CODE

3

– deviations from guideline include photoperiod and lack of detailed information

REFERENCE

Dobbs, M.G., D.S. Cherry, J.C. Scott, and J.C. Petrille. 1995. Environmental Assessment of an Alkyl

Dimethyl Benzyl Ammonium Chloride (ADBAC) Based Molluscicide Using Laboratory Tests.

Published data. Proceedings of the Fifth International Zebra Mussel and Other Aquatic Nuisance

Organisms Conference, Toronto, Canada.

(EPA

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### CHRONIC TOXICITY TO AQUATIC ORGANISMS

Title: Daily Static

- Renewal Early Life Stage Toxicity Test of Alky l Dimethyl Benzyl Ammonium

Chloride (ADBAC) to Fathead Minnows. Project No. SC890057

The study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: ADBAC, CAS No. 68424

- 85

- 1

Remarks: 30% ai

METHOD

Method/guidelin e followed: OPPTS 850.1400

GLP: Not stated

Year study performed: Not stated

Test type: Static

- renewal

Species/strain/supplier: Fathead minnow ( Pimephales promelas )

Analytical monitoring: Not stated

Exposure period: 34 days

Number of animals/sex /dose: Not stated

Statistical methods used: Not stated

Method remarks: Early life stage test

RESULTS

Nominal concentrations: Not stated

Measured concentrations: Not stated

Element value: LOAEC = 0.0759 mg/L a.i.; NOAEC = 0.0322 mg/L a.i.; MATC = 0.0494 a.i.

Remarks: Not stated

CONCLUSIONS

None stated.RELIABILITY CODE

2

REFERENCES

MRID 423021

- 02 . McIntyre, D.O. and H.O. Pate. 1992. Daily Static

- Renewal Early Life Stage Toxicity

Test of Alkyl Dimethyl Benzyl Ammonium Chloride (ADBAC) t o Fathead Minnows. Project No.

SC890057. Unpublished data. Conducted by Battelle Columbus Operations for ADBAC Quat Joint

Venture/Chemical Specialties Manufacturers Assocation.

(EPA

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- 2006

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- 0021[1])Title: Daily Static

- Renewal Chronic 21

- day Toxicity Test of Alkyl Dimethyl Benzyl Ammonium

Chloride (ADBAC) to Daphnia magna . Study No. SC890056

The study is: Adequate Not Adequate

This study was considered inadequate due to high mortality in the control group.

TEST SUBSTANCE

Identity: ADBAC, CAS No. 68424

- 85

- 1

Remarks: 3 0% ai

METHOD

Method/guideline followed: OPPTS 850.1300

GLP: Not stated

Year study performed: Not stated

Test type: Static

- Renewal

Species/strain/supplier: Water flea ( Daphnia magna )

Analytical monitoring: Not stated

Exposure period: Not stated

Number of animals/sex/dose: Not stated

Statistical methods used: Not stated

Method remarks: Numerous deviations from guideline include MATC not established

RESULTS

Nominal conce ntrations: Not stated

Measured concentrations: Not stated

Element value: LOAEC = Not determined; NOAEC 0.00415 mg/L a.i.; MATC = Not Determined

Remarks: Not stated

CONCLUSIONS

None stated.

RELIABILITY CODE3

- High control mortality

REFERENCES

MRID 423021

- 01. McIntyre, D.O. and H.O. Pate. 1992. Daily Static

- Renewal Chronic 21

- day Toxicity

Test of Alkyl Dimethyl Benzyl Ammonium Chloride (ADBAC) to Daphnia magna . Study No.

SC890056. Unpublished data. Conducted by Batalle Columbus Division fo r ADBAC Quat Joint

Venture/Chemical Specialties Manufacturers Assocation.

(EPA

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### ACUTE TOXICITY STUDIES

Summary of endpoint: Adequate

Title : An Acute Oral Toxicity Study in Rats with BQ 451

- 8: Lab Project Number: 3192.32

The study is: Adequate Not Adequate

TEST SUBSTANCE

Identity : ADBAC , CAS No. 68424

- 85

- 1

Remarks : Purity 82.26%

METHOD

Method/guideline followed: 870.1100

Type: Acute oral

GLP : Not stated

Year : Not stated

Species/Strain: Rat

Sex: Both

No. of animals per sex per dose: Not stated

Vehicle : Not stated

Route of administration: Oral

Method remarks:

Females: 0, 100, 250, 500 or 510 mg/kg

Males: 0, 500, 510, 525, 600 or 750 mg/kg

Rats (5/sex/dose)

RESULTS

L D50 value:

304.5 mg/kg (both sexes combined)

510.9 mg/kg (males)

280.8 mg/kg (females)

Number of deaths at each dose level: Two 200 mg/kg females, four 500 mg/kg females, three males,and four females in the 510 mg/kg group and all males in the 525, 600, and 750 mg/kg groups were dead

by Day 6.

Remarks: Two 200 mg/kg females, four 500 mg/kg females, three males, and four females in the 510

mg/kg group and all males in the 525, 600, and 750 mg/kg groups were dead by Day 6. Breathing

abnormalities, decrea sed activity, wobbly gait, salivation, apparent hypothermia, soft/mucoid stools,

diarrhea, decreased/no defecation, fecal/urine stain, piloerection, rough haircoat, hunched postures,

dehydration, ocular discharge, decreased food consumption, and/or dark ma terial around the facial area

were noted from most of the rats with recovery by Day 13 or earlier. Two surviving 510 mg/kg males

lost weight during the first week of study, but gained weight during the second week. All other surviving

rats gained weight during the study. The decedents had abnormal content and/or reddened mucosa in the

digestive tract, dark red thymus, and/or fluid contents in the thoracic cavity, abnormal content in the small

intestines and/or thickened stomach mucosa.

CONCLUSIONS

Tox ic Category II

RELIABILITY CODE

2

REFERENCES

45109204 (MRID) Bonnette, K. (1998) An Acute Oral Toxicity Study in Rats with BQ 451

- 8: Lab

Project Number: 3192.32. Unpublished study prepared by Springborn Laboratories. 90 p.

(EPA

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- 2006

- 0339

- 0 019[1]; p. 3)Title : Acute Dermal Toxicity Study in Rats: Albemarle BQ451

- 8 Biocide: Lab Project Number:

7466:P322

The study is: Adequate Not Adequate

TEST SUBSTANCE

Identity : ADBAC , CAS No. 68424

- 85

- 1

Remarks : Purity 82.26%

METHOD

Method/guideline followed: 870.1200

Type: Acute dermal

GLP : Not stated

Year : Not stated

Species/Strain: Rat

Sex: Both

No. of animals per sex per dose: Not stated

Vehicle : Not stated

Route of administration: Dermal

M ethod remarks:

0, 500, 1000 or 2000 mg/kg

Rats (5/sex/dose)

RESULTS

LD50 value:

930 mg/kg (combined)

1100 mg/kg (males)

704 mg/kg (females)

Number of deaths at each dose level: Five 2000 mg/kg females were found dead on Day 1. One 500

mg/kg female, one male and four females in the 1000 mg/kg group, and five 2000 mg/kg were found dead

by Day 4.

Remarks: Five 2000 mg/kg females were found dead on Day 1. One 500 mg/kg female, one male and

four females in the 1000 mg/kg group, and five 2000 mg/kg were found dead by Day 4. Irregular

respirations, hunched postures, hypoactivity, coldness to the touch, reduced fecal volume, tremors, ataxia,and/or dyspnea were noted for the decendents prior to death. Hypoactivity, hunched postures, reduced

fecal volume, and/or irregular respiration were noted for some of the surviving rats with recovery by Day

2. Erythema and edema were noted at dose site from most of the rats. Eschar was also noted at the dose

site from surviving rats. All surviving rats had normal bo dy weight gains. No gross abnormalities were

noted from the surviving rats and one female decedent in the 1000 mg/kg group. The other decedents had

slightly/moderately red lungs, discolored livers, and/or red/black/green/yellow intestines.

CONCLUSIONS

Toxic Category II

RELIABILITY CODE

2

REFERENCES

45109202 (MRID) Moore, G. (1999) Acute Dermal Toxicity Study in Rats: Albemarle BQ451

- 8 Biocide:

Lab Project Number: 7466:P322. Unpublished study prepared by Product Safety Labs. 25p.

(EPA

- HQ

- OPP

- 2006

- 0339

- 0019[1])Title : Acute Inhalation Toxicity Study in Rats: Albemarle BQ451

- 8 Biocide: Lab Project Number: 7467:

P330

The study is: Adequate Not Adequate

TEST SUBSTANCE

Identity : ADBAC , CAS No. 68424

- 85

- 1

Rem arks : Purity 82.26%

METHOD

Method/guideline followed: 870.1300

Type: Acute inhalation

GLP : Not stated

Year : Not stated

Species/Strain: Rat

Sex: Not stated

No. of animals per sex per dose: Not stated

Vehicle : Not stated

Route of administration: I nhalation

Method remarks:

0.054 or 0.51 mg/L

Rats (5/sex/dose)

RESULTS

LC 50 value: 0.054 < LC50 < 0.51 mg/L

Number of deaths at each dose level: 0.054 mg/L group: All animals survived; 0.51 mg/L group: All

animals died.

Remarks: 0.054 mg/L group: All animals survived. Animal displayed ocular and nasal discharge,

irregular respiration, dyspnea, hunched posture, and hypoactivity. Irregular respiration and/or

hypoactivity persisted in several rats upon removal from the exposure chamber and some anima ls also

developed facial staining or reduced fecal volume. All rats recovered from the above symptoms by Day 2

and appeared active and healthy for the remainder of the study, gaining bodyweight over the entire 14

- day

observation period. No gross abnormal ities were noted in animals when necropsied.0.51 mg/L group: All animals died. During exposure, animals exhibited ocular and nasal discharge,

irregular respiration, dyspnea, gasping, hunched posture, and hypoactivity prior to death. Gross necropsy

of o ne animal revealed discoloration and edema of the lungs. Gross abnormalities were not noted in the

other animals.

CONCLUSIONS

Toxicity Category II

RELIABILITY CODE

2

REFERENCES

44885201 (MRID) Wnorowski, G. (1999) Acute Inhalation Toxicity Study i n Rats: Albemarle BQ451

- 8

Biocide: Lab Project Number: 7467: P330. Unpublished study prepared by Product Safety Labs. 30 p.

(EPA

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### SKIN IRRITATION STUDY

Title: Primary Skin Irritation Study in Rabbits: Albemarle BQ451

- 8 Bioci de: Lab Project Number: 7468:

P326

TEST SUBSTANCE

Identity: ADBAC , CAS No. 68424

- 85

- 1

Remarks: Purity 82.26%

METHOD

Method/guideline followed: 870.2500

GLP (Y/N): Not stated

Species/strain: Rabbit

Sex (males/females/both): Female

Number of animal s/sex/dose: Not stated

Volume administered: 0.5 mL BQ451

- 8, 40 hours

Test condition: Not stated

RESULTS

Corrosive . The rabbit had well

- defined erythema one hour after patch removal. The erythema intensified

to severe by 24 hours and persisted through 72 hours. Moderate edema was noted in the rabbit one hour

after patch removal, but reduced to slight by 24 hours through 72 hours . The application site showed

superficial eschar by 24 through 48 hours and signs of dermal corrosion by 72 hours. The prima ry dermal

irritation index was 5.8.

CONCLUSIONS

Toxic Category I

RELIABILITY CODE

2

REFERENCES

45109201 (MRID) Moore, G. (1999) Primary Skin Irritation Study in Rabbits: Albemarle BQ451

- 8

Biocide: Lab Project Number: 7468: P326. Unpublished stud y prepared by Product Safety Labs. 15 p.

(EPA

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### SKIN SENSITIZATION STUDY

Title: Dermal Sensitization Study in Guinea Pigs (Buehler Method): Albermarle BQ451

- 8 Biocide: Lab

Project Number: 7469: P328

TEST SUBSTANCE

Identity: ADBAC , CAS No. 68424

- 85

- 1

Remarks: Purity 82.26%

METHOD

Method/guideline followed: 870.2600

GLP (Y/N): Not stated

Species/strain: Guinea pigs

Sex (male/female/both): Both

Number of animals/sex/dose: 20 test animals, 10 controls; 24 males, 6 female s

Dose/concentration: Induction: 0.4 mL of 2% w/w test material in distilled water with occlusive 25 mm

Hilltop Chamber for 6 hours once/week, 3 weeks.

Challenge Period: 27 days

Challenge dose: 0.4 mL of 0.5% w/w test material in distilled water under occ lusion for 6 hours.

Control: 0.4 mL of 0.5% w/w test material in distilled water at challenge only

Test condition: Not stated

RESULTS

Faint usually confluent to moderate erythema was noted on all animals after the first induction. Very

faint usually n onconfluent to moderate erythema was noted on 19/20 animals and severe erythema was

noted on 1/20 animals after the second induction. Faint usually confluent to severe erythema was noted

on all animals after the third induction. Eschar was noted on 2/20 animals 48 hours after the third

induction. Desquamation was noted on one animal 24 hours after the third induction with resolution by

48 hours. After three weekly inductions, dermal irritation suggestive of sensitization was not observed on

the test ani mals after challenge. The study report included DNCB positive control study which was

carried out within six months of the current study. The results were appropriate.

CONCLUSIONS

Not a dermal sensitizer.

RELIABILITY CODE

2REFERENCES

45109203 (MRI D) Moore, G. (1999) Dermal Sensitization Study in Guinea Pigs (Buehler Method):

Albermarle BQ451

- 8 Biocide: Lab Project Number: 7469: P328. Unpublished study prepared by Product

Safety Labs. 24 p.

(EPA

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### GENETIC TOXICITY ST UDY

Summary of endpoint: Adequate

Title: Mutagenicity Test on Alkyl Dimethyl Benzyl Ammonium Chloride: In the CHO/HGPRT Forward

Mutation Assay: HLA Study No. 10238

- 0

- 435

The study is: Adequate Not Adequate

TEST S UBSTANCE

Identity: ADBAC , CAS No. 68424

- 85

- 1

Remarks : 80% purity

METHOD

Method/guideline followed: 870.5300

Test Type: HGPRT Assay in CHO cells for forward mutation

Test system: Mammalian cell, in vitro

GLP (Y/N): Not stated

Year study performed: Not stated

Species/Strain/cell

- type/cell line: Chinese Hamster ovary cells

Metabolic activation: Not stated

Concentrations tested: 0, 1.0, 2.0, 5.0, 10.0, 20.0, 50.0, 100.0, 200.0, 500.0 or 5000.0 μg/mL, with and

without metabolic activation

Remarks : None

RESULTS

Overall results: Negative

Cytotoxic concentration: Not stated

Remarks: Mutant frequencies of all cultures treated with the test material were within the acceptable

range for background mutant frequencies (0 to 15 x 10

- 6 ). In all of the assays, no culture had a mutant

frequency that was statistically elevated over background levels and greater than 15 x 10

- 6 . Three toxic

cultures in one nonactivation trial did achieve statistical significance. The significant mutant frequencies

were wi thin the acceptable background range and were not confirmed at equitoxic dose levels I the

independent nonactivation trial. The significant cultures, in that one trial were apparently due to normalassay variation. ADBAC was considered negative for induci ng forward mutations at the HGPRT locus in

CHO cells under the S9 metabolic activation and nonactivation conditions of this assay.

CONCLUSIONS

A D B AC showed negative evidence of mutagenicity in Chinese Hamster ovary cells.

RELIABILITY CODE

2

REFERENCE

41012701 (MRID) Young, R. (1989) Mutagenicity Test on Alkyl Dimethyl Benzyl Ammonium

Chloride: In the CHO/HGPRT Forward Mutation Assay: HLA Study No. 10238

- 0

- 435. Unpublished

study prepared by Hazleton Laboratories America, Inc. 67 p.

(EPA

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### CHROMOSOMAL ABERRATIONS STUDY

Summary of endpoint: Adequate

Title: Assessment of the Mutagenic Activity of Hya

- mine 3500 in the Mouse Micronucleus Test: Lab.

No. 10753

The study is: Adequate No t Adequate

TEST SUBSTANCE

Identity: ADBAC , CAS No. 68424

- 85

- 1

Remarks: 80% purity

METHOD

Method/guideline followed: 870.5385

Test Type: In vivo bone marrow chromosome aberration assay

Test system: In vivo

GLP (Y/N): Not stated

Year study perfor med: Not stated

Species/Strain/cell

- type/cell line: mouse bone marrow cells

Metabolic activation: Not stated

Concentrations tested: Single dose of 400 mg/kg, sacrificed at 24, 48 or 72 hours later

Remarks:

Mice (5/sex/group)

Solvent control: deioniz ed water

Positive control: Cyclosphosphamide (CP, 30 mg/kg)

RESULTS

Overall results: Negative

Cytotoxic concentration: Not stated

Remarks: One mouse of the 72

- hour group was found dead on the third day after treatment. The

PCE/NCE ratio was reduced i n all test groups (as well as the positive controls), but no alteration in MN

from solvent control value was found in any timed test group. By contrast, the CP

- group evidenced a

significant increase in MN

- PCE.CONCLUSIONS

A D B AC showed negative evidence o f mutagenicity in an in vivo bone marrow chromosome aberration

assay

.

RELIABILITY CODE

2

REFERENCE

40311101 (MRID) Kallerson, T. (1985) Assessment of the Mutagenic Activity of Hya

- mine 3500 in the

Mouse Micronucleus Test: Lab. No. 10753. Un

- published study prepared by Scantox Labs Ltd. 17p

43037701 (MRID) Schoenig, G. (1993) Response to EPA Data Evaluation Review for Study Entitled:

“Chromosome Aberrations in vivo

— Mouse Micronucleus Test”: Lab Project Number: 10753.

Unpublished study prepared by SC ANTOX Biologisk Laboratorium A/S. 7 p.

(EPA

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- 0019[1])Title: Alkyl Dimethyl Benzyl Ammonium Chloride (ADBAC) in the Rat Primary Hepatocyte

Unscheduled DNA Synthesis Assay: An addendum: Lab Project Number: 10238

- 0

- 447

TEST SUBSTANCE

Iden tity: ADBAC , CAS No. 68424

- 85

- 1

Remarks: 80% purity

METHOD

Method/guideline followed: 870.5550

Test Type: Unscheduled DNA synthesis assay

Test system: Mammalian cells

GLP (Y/N): Not stated

Year study performed: Not stated

Species/Strain/cell

- typ e/cell line: Rat hepatocyte cells

Metabolic activation: Not stated

Concentrations tested: Tritiated thymidine (38Ci/mmol) for 18.8 hours together with 0.538, 1.08, 2.15,

4.31, 5.38, 6.46, 7.00, 7.54, 8.07 or 8.61 μg/mL ADBAC

Remarks:

Solvent control: deionized water

Positive control: AAF, 0.10 μg/mL

RESULTS

Overall results: Negative

Cytotoxic concentration: Not stated

Remarks: The test article was lethal at 8.61 ug/mL, and progressively cytotoxic at doses equal to or

above 4.31 ug/mL. Treatments producing moderate levels of cytotoxicity (62.7% relative survival and

greater) as well as non

- toxic doses were analyzed for UDS. None of these analyzable treatments induced

silver grain labeling different from solvent control. In contrast, the mutagen 2

- AAF induced large

increases in nuclear labeling in the absence of significant toxicity.

CONCLUSIONS

ADBAC showed negative evidence of mutagenicity in an unscheduled DNA synthesis assay.

RELIABILITY CODE2

REFERENCE

42290802 (MRID) McKeon, M. (1989) Alkyl Dimethyl Benzyl Ammonium Chloride (ADBAC) in the

Rat Primary Hepatocyte Unscheduled DNA Synthesis Assay: An addendum: Lab Project Number:

10238

- 0

- 447. Unpublished study prepared by Hazleton Laboratories America, Inc. 11 p.

42290801 (MRID) McKeon, M. (1992) Genotoxicity Test on Alkyl Dimethyl Benzyl Ammonium

Chloride (ADBAC) in the Assay for Unscheduled DNA Synthesis in Rat Liver Primary Cell Cultures:

Lab Project Number: 14778

- 0

- 447. Unpublished study prepared by Hazleton Washington, Inc. 51 p.

(EPA

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### REPEATED

- DOSE TOXICITY STUDY

Summary of endpoint: Adequate

Title: 21

- Day Dermal Toxicity

– Guinea Pigs, OPPTS 870.3200

The study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: formulated product

Remarks: 6% ai; ADBAC , CAS No. 68424

- 85

- 1

, contained within HS

- sanitizing carpet shampoo at a 1:5 dilution

METHOD

Method/guideline followed: OPPTS 870.3200

GLP: Not stated

Year study was performed: Not stated

Test type: 21

- day d ermal toxicity test

Species/strain: Guinea pig (strain not specified)

Sex: Both

Route of administration: Dermal

Doses: 1000 mg/kg/day of ADBAC

Exposure period: 24 hours

Frequency of treatment: 5 days per week for 3 weeks

Control group and treatmen t: Not stated

Post

- observation period: Not stated

Statistical methods: Not stated

Method remarks: In a 21

- day dermal toxicity study, a dose of 1000 mg/kg/day (limit dose) of ADBAC,

contained within HS

- sanitizing carpet shampoo at a 1:5 dilution, was t ested in male and female guinea

pigs, 5 days per week, for 3 weeks. Each exposure was for 24 hours to groups of 5 guinea pigs/sex/dose.

RESULTS

NOAEL and LOAEL:LOAEL (systemic) = 1000 mg/kg/day based on decreases in body weight in female pigs

NOAEL (systemic) < 1000 mg/kg/day (not achieved)

LOAEL (dermal) = 1000 mg/kg/day based on denuding of the epidermal layer

NOAEL (dermal) not achieved

Actual dose received by dose level by sex: Not stated

Remarks: Over the course of the study, body weight los ses of between 6

- 11% were observed in male and

female guinea pigs. There were no reported mortalities or clinical signs of toxicity. Slight reduction in

food consumption (2

- 5%) was observed over the course of this study. There were no significant

treatm ent

- related effects on hematology or clinical chemistry. Histological examination of the skin

showed that the non

- vascularized epidermal layer was denuded, while the dermal or corium layer and

basement membrane appeared normal. There were no treatment

- re lated effects reported on the liver and

kidney.

CONCLUSIONS

The Systemic LOAEL = 1000 mg/kg/day, based on decreases in body weight in female guinea pigs. The

Systemic NOAEL < 1000 mg/kg/day (not achieved). The Dermal LOAEL = 1000 mg/kg/day, based on

de nuding of the epidermal layer. The Dermal NOAEL was not achieved.

RELIABILITY CODE

2

- The study contained several experimental deficiencies but had enough information on dermal effects to

be used for the dermal endpoint determination

REFERENCES

407007 00 (MRID)

(EPA

- HQ

- OPP

- 2006

- 0339

- 0019[1])Title: Acute Toxicology (EP): HS

- Sanitizing Carpet Shampoo: Project ID: B6

- 27

The study is: Adequate Not Adequate

This study was considered inadequate due to testing at a non

- standard duration (< 28

- days).

TEST SUBSTANCE

Identity: ADBAC, CAS No. 68424

- 85

- 1

Remarks: 81% ai

METHOD

Method/guideline followed: OPPTS 870.3200

GLP: Not stated

Year study was performed: Not stated

Test type: 21

- day dermal toxicity test

Species/strain: Guinea pig (strain not specified)

Sex: Both

Route of administration: Dermal

Doses: 0, 500 , and 1000 mg/kg /day

Exposure period: 24 hours

Frequency of treatment: 5 days per week for 3 weeks

Control group and treatment: Not stated

Pos t

- observation period: Not stated

Statistical methods: Not stated

Method remarks: A 1:5 dilution of HS

- sanitizing carpet shampoo (containing 6% didecyl ammonium

chloride and 4% alkyl dimethyl benzyl ammonium chloride) was applied to a 2 inch square of s haved

dorsal trunk of 5 male and 5 female guinea pigs at 500 mg/kg, five days a week, for 21 days. Each

exposure was for 24 hours.

RESULTS

NOAEL and LOAEL:

Systemic toxicity:NOAEL = 500 mg/kg/day

LOAEL = 1000 mg/kg/day, based on decreases in body wei ght in females

Dermal toxicity:

NOAEL = 500 mg/kg/day

LOAEL = 1000 mg/kg/day, based on denuding of the epidermal layer

Actual dose received by dose level by sex: Not stated

Remarks: Signs of skin irritation were noted during the second week of treatment and the report stated

that the response intensified during the third week of treatment. Body weight was decreased in treated

males and females by 7% and 11% vs. untreated animals at week 3 at 1000 mg/kg. Results of

hematology and clinical chemistry meas urements indicated a slight elevation of basophils and eosinophils

as well as a slight elevation of SGPT and SGOT but statistics were not performed on these data.

Histologically, the skin irritation was described as denuded non

- vascularized epidermal laye r at the

application site.

CONCLUSIONS

The 500 mg/kg/day value could be considered a provisional NOAEL as slight effects were still observed

on the skin and the report did not represent the data on this effect at this dose but it likely that a true

NOA EL would not be vary far below this level for dermal effects.

RELIABILITY CODE

3

- This study also contained several deficiencies including lack of several experimental details (no

information on dose solution, stability and homogeneity), and missing indi vidual animal data for body

weight, clinical observations, hematology, clinical chemistry, organ weights, gross pathology and

histopathology.

REFERENCES

41105801 (MRID) Rose, G. (1989) Acute Toxicology (EP): HS

- Sanitizing Carpet Shampoo: Project ID:

B 6

- 27. Unpublished study prepared by Envirocon. 36 p.

40565301 (MRID)

(EPA

- HQ

- OPP

- 2006

- 0339

- 0019[1])Title: Ninety

- day Subchronic Dermal Toxicity Study with Alkyl Dimethyl Benzyl Ammonium Chloride

(ADBAC) in Rats: Lab Project I.D.: 52

- 623

The stud y is: Adequate Not Adequate

TEST SUBSTANCE

Identity: ADBAC, CAS No. 68424

- 85

- 1

Remarks: 81.09% a.i.

METHOD

Method/guideline followed: 870.3250

GLP: Not stated

Year study was performed: Not stated

Test type: 9 0

- day dermal toxicity study

Species/strain: Sprague

- Dawley rats

Sex: Both

Route of administration: dermal (to clipped backs)

Doses: 0, 2, 6 or 20 mg/kg/day (as 0.1, 0.3 and 1.0% solutions at a dose volume of 2 ml/kg)

Exposure period: 6 to 8 hours

Fre quency of treatment: 5 days per week for 13 weeks.

Control group and treatment: a control was included (0 mg/kg/day)

Post

- observation period: Not stated

Statistical methods: Not stated

Method remarks: In a 90

- day dermal toxicity study, ADBAC (81.09% a.i.) was applied at dose levels of

0, 2, 6 or 20 mg/kg/day (as 0.1, 0.3 and 1.0% solutions at a dose volume of 2 ml/kg) to the clipped backs

of Sprague

- Dawley rats for 6

- 8 hours per day, 5 days per week, for 13 weeks. Although higher doses

were tested in a preliminary range

- finding study (6, 20, 60, 120 and 200 mg/kg/day), the high dose

selected for the main study (20 mg/kg/day) was chosen on the basis that higher concentrations produced

skin irritation that was considered greater than slight.

RESULTS

N OAEL and LOAEL:

NOAEL (systemic and dermal) = 20 mg/kg/dayActual dose received by dose level by sex: Not stated

Remarks: A significant and dose

- related decrease in reticulocyte count was observed in the 6

- and 20

- mg/kg/day females. Decreases in retic ulocyte count are normally associated with regenerative responses

to anemia. However, no evidence of anemia was seen in other hematological parameters. Furthermore,

the decreased levels in treated females were similar to the levels observed in control ma les. Thus, the

decreasing reticulocyte count was most likely not a biologically significant finding.

A significant increase in hyperkeratosis was observed in treated skin of high dose females, but this lesion

was also observed in increased incidence in m ale rats at all doses including controls.

CONCLUSIONS

The NOAEL for systemic and dermal effects was determined to be 20 mg/kg/day

RELIABILITY CODE

2

REFERENCES

41499601 (MRID) Gill, M.; Wagner, C. (1990) Ninety

- day Subchronic Dermal Toxicity Study wi th

Alkyl Dimethyl Benzyl Ammonium Chloride (ADBAC) in Rats: Lab Project I.D.: 52

- 623. Unpublished

study prepared by Union Carbide Bushy Run Research Center. 264 p.

(EPA

- HQ

- OPP

- 2006

- 0339

- 0019[1])Title: Ninety

- day Dietary Toxicity Study with Alkyl Dim ethyl Benzyl Ammonium Chloride (ADBAC)

in Rats: Project ID:51

- 503.

The study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: Alkyl dimethyl benzyl ammonium chloride ( ADBAC) , CAS No. 68424

- 85

- 1

Remarks: 79.7% a .i.

METHOD

Method/guideline followed: 870.3100

GLP: Not stated

Year study was performed: Not stated

Test type: Subchronic oral toxicity

Species/strain: Rats

Sex: Both

Route of administration: Dietary

Doses: 0, 100, 500, 1000 (0, 6.3, 31.2 and 62.0 mg/kg/day for males and 0, 7.9, 38.3 and 76.7 mg/kg/day

for females), 4000 or 8000 ppm

Exposure period: 13 weeks

Frequency of treatment: Daily

Control group and treatment: 0 ppm

Post

- observation period: Not stated

Statistical methods: Not stated

Method remarks: In a subchronic oral toxicity study in rats, male and female rats (15/sex/dose) were

administered ADBAC (79.7% a.i.) in the diet for 13 weeks at dose levels of 0, 100, 500, 1000, 4000 or

8000 ppm.

RESULTS

NOAEL and LOAEL:

NOA EL (systemic; males) = 500 ppm (31.2 mg/kg/day)

LOAEL (systemic; males) = 1000 ppm (62.0 mg/kg/day) based on decreased weight gainNOAEL (systemic; females) = 1000 ppm (76.7 mg/kg/day)

LOAEL (systemic; females) = 4000 ppm (>76.7 mg/kg/day) based on increas ed mortality, decreased

body weight gain and food consumption and increased lesions in females

Actual dose received by dose level by sex: 0, 6.3, 31.2 and 62.0 mg/kg/day for males and 0, 7.9, 38.3

and 76.7 mg/kg/day for females for 0, 100, 500 and 1000 pp m, respectively. Increased mortality at the

two highest dose levels did not allow for daily compound consumption to be calculated.

Remarks: Increased mortality was observed at the 4000 and 8000 ppm dose groups in both sexes.

Decreased body weight, weigh t gain, food consumption and increased incidence of microscopic lesions

[congestion and edema of the G.I. tract, hemorrhaging of the lungs and brain] were also observed in males

and females at 4000 ppm ]

.

CONCLUSIONS

The systemic NOAEL for males is 500 ppm and for females is 1000 ppm, based upon decreased body

weight and weight gain in males at 1000 ppm, and increased mortality, decreased body weight gain and

food consumption, and increased lesions in females at 4000 ppm.

RELIABILITY CODE

2

– This stud y is classified as acceptable.

REFERENCES

40746601 (MRID) Van Miller, J.; Weaver, E. (1998) Ninety

- day Dietary Toxicity Study with Alkyl

Dimethyl Benzyl Ammonium Chloride (ADBAC) in Rats: Project ID:51

- 503. Unpublished study

prepared by Bushy Run Re

- sea rch Center. 300p.

(EPA

- HQ

- OPP

- 2006

- 0339

- 0019[1])Title: Evaluation of ADBAC in a One

- Year Chronic Dietary Toxicity Study in Dogs: Lab Project

Number: 638

- 004

The study is: Adequate Not Adequate

TEST SUBSTANCE

Ide ntity: ADBAC, CAS No. 68424

- 85

- 1

Remarks: 81% purity

METHOD

Method/guideline followed: 870.4100

GLP: Not stated

Year study was performed: Not stated

Test type: Chronic toxicity

Species/strain: Beagle dogs

Sex: Both

Route of administration: di et

Doses: 0, 3.79, 13.1 or 33.8 mg/kg/day in males and 0, 3.67, 14.6 or 38.6 mg/kg/day in females (1, 120,

400 or 1200 ppm)

Exposure period: one year

Frequency of treatment: daily

Control group and treatment: 0 mg/kg/day

Post

- observation period: Not stated

Statistical methods: Not stated

Method remarks: In a chronic toxicity study in dogs, groups of 4 male and female beagle dogs per

group received either 0, 120, 400 or 1200 ppm (0, 3.79, 13.1 or 33.8 mg/kg/day in males and 0, 3.67, 14.6

or 38.6 m g/kg/day in females) alkyl dimethyl benzyl ammonium chloride [ADBAC, 80% a.i.] as a direct

dietary admix for one year.

RESULTS

NOAEL and LOAEL:NOAEL (m/f) = 3.79 / 3.67 mg/kg/day

LOAEL (m/f) = 13.1 / 14.6 mg/kg/day based on reduced body weight

Ac tual dose received by dose level by sex: Not stated

Remarks: Systemic toxicity was observed at 400 ppm and above in female dogs and at 1200 ppm in

males as reduced body weight gain (approximately 10% reduction) and 52 weeks of exposure. Food

consumption was decreased in the 1200 ppm males and females for the entire study period

(approximately 15% reduction in males and 5% reduction in females).

CONCLUSIONS

Based on the data in this study, the Systemic Toxicity NOAEL was 120 ppm (3.79 mg/kg/day in male s,

3.67 mg/kg/day in females) and the LOAEL was 400 ppm (13.1 mg/kg/day in males, 14.6 mg/kg/day in

females) based on reduced body weight.

RELIABILITY CODE

2

REFERENCES

43221101 (MRID) Goldenthal, E. (1994) Evaluation of ADBAC in a One

- Year Chronic Di etary Toxicity

Study in Dogs: Lab Project Number: 638

- 004. Unpublished study prepared by International Research and

Development Corp. 355 p.

(EPA

- HQ

- OPP

- 2006

- 0339

- 0019[1])

### CARCINOGENICITY STUDIES

Title: Chronic Dietary Oncogenicity Study with Alkyl Dimethyl Benzyl Ammonium Chloride (ADBAC)

in Mice: Lab Project Number: 53

- 515

The study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: Alkyl dimethyl benzyl ammonium chloride ( ADBAC) , CAS No. 68424

- 85

- 1

Remar ks: 81% purity

METHOD

Method/guideline followed: 870.4200

GLP: Not stated

Year study was performed: Not stated

Test type: Carcinogenicity

Species/strain: CD

- 1 mice

Sex: 60/sex/dose

Route of administration: diet

Doses: nominal doses of 14.9, 7 3.4 and 229.3 mg/kg/day in males; 17.8, 92.1 and 288.6 mg/kg/day in

females (0, 100, 500, or 1500 ppm)

Exposure period: 78 weeks

Frequency of treatment: daily

Control group and treatment: 0 mg/kg/day

Post

- observation period: Not stated

Statistical m ethods: Not stated

Method remarks: In a carcinogenicity study in mice, ADBAC (81% purity) was administered in the diet

to male and female CD

- 1 (60/sex/dose) at levels of 0, 100, 500 or 1500 ppm for 78 weeks (nominal doses

of 14.9, 73.4 and 229.3 mg/kg/d ay in males; 17.8, 92.1 and 288.6 mg/kg/day in females).

RESULTSNOAEL and LOAEL:

NOAEL (m/f) = 229.3 / 288.6 mg/kg/day

LOAEL (m/f) = 73.4 / 92.1 mg/kg/day , based on decreased body weight

Actual dose received by dose level by sex: Not stated

Remar ks: No significant differences in the incidence of mortality were observed in treated animals

versus controls. No clinical signs of toxicity were observed at any dose level tested. Significant

reductions in group mean body weight were observed at the hig h dose in male and female mice

throughout the treatment period with no significant reduction in food intake. There were no significant

treatment

- related effects on organ weights, macroscopic or microscopic pathology in treated mice at any

dose level.

C ONCLUSIONS

ADBAC was negative for carcinogenicity in this study. The Systemic LOAEL = 1500 ppm in male and

female mice (229.3 / 288.6 mg/kg/day), based on reduced body weight. The Systemic NOAEL = 500

ppm in male and female mice (73.4 / 92.1 mg/kg/day).

RELIABILITY CODE

2

REFERENCES

41765201 (MRID) Gill, M.; Hermansky, S.; Wagner, C. (1991) Chronic Dietary Oncogenicity Study

with Alkyl Dimethyl Benzyl Ammonium Chloride (ADBAC) in Mice: Lab Project Number: 53

- 515.

Unpublished study prepared by Bush y Run Research Center. 1083 p.

(EPA

- HQ

- OPP

- 2006

- 0339

- 0019[1])Title: Chronic Dietary Toxicity/Oncogenicity Study with Alkyl Dimethyl Benzyl Ammonium Chloride

(ADBAC) in Rats: Lab Project Number: 53

- 543

The study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: Alkyl dimethyl benzyl ammonium chloride ( ADBAC) , CAS No. 68424

- 85

- 1

Remarks: 81% purity

METHOD

Method/guideline followed: OPPTS 870.4300

GLP: Not stated

Year study was performed: Not stated

Test type: Chronic Toxicity / Carcinogenicity in Rats

Species/strain: Sprague

- Dawley rats

Sex: Both

Route of administration: Dietary

Doses: 0, 300, 1000 or 2000 ppm (nominal doses of 13, 44 and 88 mg/kg

- bw/day in males; 17, 57 and

116 mg/kg

- bw/day in females)

Exposure period: 104 weeks

Frequency of treatment: Daily

Control group and treatment: a control group was included (0 mg/kg

- bw/day)

Post

- observation period: Not stated

Statistical methods: Not stated

Method remarks: In a chronic toxic ity / carcinogenicity study, ADBAC (81% purity) was administered

in the diet to groups of male and female Sprauge

- Dawley rats (50/sex/dose) at dose levels of 0, 30, 1000

and 2000 ppm (nominal doses of 13, 44 and 88 mg/kg/day in males; 17, 57 and 116 mg/kg/ day in

females) for 104 weeks.

RESULTS

NOAEL and LOAEL:NOAEL (m/f) = 44 / 57 mg/kg

- bw/day

LOAEL (m/f) = 88 / 116 mg/kg

- bw/day based on decreased body weight and weight gain

Actual dose received by dose level by sex:

Remarks: Significant decreas es in group mean body weight were observed in male rats at the 2000 ppm

dose level during weeks 1

- 26 of the study and then sporadically thereafter. Body weights of high dose

female rats were also significantly decreased during weeks 1

- 60 of the study. Bo dy weight gain was

decreased 11% on average in high dose males and 14% in high dose females. There were no significant

treatment

- related effects on clinical chemistry, hematology, or urinalysis. No treatment

- related non

neoplastic gross or microscopic le sions were evident in any of the treated groups of rats. There was no

evidence of carcinogenicity of ADBAC in this study.

CONCLUSIONS

The systemic toxicity NOAEL = 1000 ppm, (44 mg/kg/day [M]; 57 mg/kg/day [F]), and the Systemic

toxicity LOAEL = 2000 p pm (88 mg/kg/day [M]; 116 mg/kg/day [F], based on decreased body weight

and weight gain.)

RELIABILITY CODE

2

REFERENCES

41947501 (MRID) Gill, M.; Hermansky, S.; Wagner, C. (1991) Chronic Dietary Toxicity/Oncogenicity

Study with Alkyl Dimethyl Benzyl A mmonium Chloride (ADBAC) in Rats: Lab Project Number: 53

- 543. Unpublished Study prepared by Bushy Run Research Center. 1671 p.

(EPA

- HQ

- OPP

- 2006

- 0339

- 0019[1])

### REPRODUCTIVE TOXICITY STUDIES

Summary of endpoint: Adequate

Title: Two

- generation Reprodu ction Study in Sprague

- Dawley (CD) Rats with Alkyl Dimethyl Benzyl

Ammonium Chloride (ADBAC) Administered in the Diet: Project Report 52

- 52

- 254: Project Nos. 87

-

37

- 97105; 87

- 37

- 97109.

The study is: Adequate Not Adeq uate

TEST SUBSTANCE

Identity: ADBAC, CAS No. 68424

- 85

- 1

Remarks: 81.09% pure

METHOD

Method/guideline followed: 870.3800

GLP: No

Year study was performed: Not stated

Species/strain: Sprague

- Dawley rats

Sex: Both

Route of administration: diet

Doses: 0, 300, 1000 or 2000 ppm. Mean compound consumption was 20.7, 68.2 and 134.7 mg/kg/day for

F0 males, and 25.5, 81.3 and 164.7 mg/kg/day for F0 females. For the F1 males, mean compound

consumption was 19.1, 62.5, and 125.4 mg/kg/day, and 24.8, 7 8.5 and 157.1 mg/kg/day for F1 females.

Exposure period: 10 weeks for F0 parents; 15 weeks for F1 parents

Frequency of treatment: daily

Control group and treatment: 0 mg/kg/day

Post

- observation period: Not stated

Statistical methods: Not stated

Met hod remarks: In a two

- generation reproductive toxicity study in rats, ADBAC (81.09%) was

administered in the diet to groups of male and female Sprague

- Dawley rats (28/sex/dose) at dose levels of

0, 300, 1000 or 2000 ppm over two generations. After 10 week s of dietary treatment, F0 parental animals

were mated. F1 parental animals were mated after 15 weeks of dietary treatment. Mean compoundconsumption was 20.7, 68.2 and 134.7 mg/kg/day for F0 males, and 25.5, 81.3 and 164.7 mg/kg/day for

F0 females. For the F1 males, mean compound consumption was 19.1, 62.5, and 125.4 mg/kg/day, and

24.8, 78.5 and 157.1 mg/kg/day for F1 females.

RESULTS

NOAEL and LOAEL:

NOAEL (Parental) = 146 mg/kg/day

LOAEL (Parental) > 146 mg/kg/day (highest dose tested)

NOAEL (d evelopmental/systemic) = 65.4/79.9 mg/kg/day (M/F)

LOAEL (developmental/systemic) = 130.1/160.9 mg/kg/day (M/F) based on reduced pup body weight

and weight gain during lactation

Remarks: There was no treatment

- related mortality in parental animals at any dose level, and there were

no reported signs of clinical toxicity in parental animals. Although some decrease in body weight was

observed in both generations at the top dose, the significant variability observed did not qualify this as a

treatment

- related effect.

In pups of both generations, mean body weights at the top dose were significantly reduced during

lactation and post

- weaning periods. There were no adverse effects noted on gestational length, mating,

fertility, or other gestational indices.

CON CLUSIONS

Based on the results of this study, the Parental NOAEL = 146 mg/kg/day, and the Parental LOAEL > 146

mg/kg/day (highest dose tested). The Developmental/Systemic NOAEL = 65.4/79.9 mg/kg/day (M/F),

and the Developmental/Systemic LOAEL = 130.1/160. 9 mg/kg/day (M/F), based on reduced pup body

weight and weight gain during lactation [doses for both the F0 and F1 pups combined].

RELIABILITY CODE

2

REFERENCES

41385001 (MRID) Neeper

- Bradley, T. (1990) Two

- generation Reproduction Study in Sprague

- Daw ley

(CD) Rats with Alkyl Dimethyl Benzyl Ammonium Chloride (ADBAC) Administered in the Diet: Project

Report 52

- 52

- 254: Project Nos. 87

- 37

- 9710 5; 87

- 37

- 97109 . Unpublished study prepared by Bushy Run

Research Center. 492 p.

(EPA

- HQ

- OPP

- 2006

- 0339

- 0019[1])

### DEVELOPMENTAL TOXICITY STUDY

Summary of endpoint: Adequate

Title: Developmental Toxicity Evaluation II of Alkyl Dimethyl Benzyl Ammonium Chloride (ADBAC)

Administered by Gavage to CD Rats: Lab Project Number: 91N0031

The study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: ADBAC, CAS No. 68424

- 85

- 1

Remarks: 81.09% ai

METHOD

Method/guideline followed: 870.3700

GLP: Not stated

Year study was performed: Not stated

Species/strain: Sprague

- Da wley rats

Sex: Female

Route of administration: gavage

Doses: 0, 10, 30 or 100 mg/kg

- bw/day

Exposure period: on gestation days 6 through 15

Frequency of treatment: Daily

Control group and treatment: a control was included (0 mg/kg

- bw/day)

Post

- obse rvation period: Not stated

Statistical methods: Not stated

Method remarks: Female Sprauge

- Dawley rats (25/dose) were administered ADBAC (81.09% a.i.) by

gavage at doses of 0, 10, 30 and 100 mg/kg on gestation days 6 through 15 inclusive for assessment o f

developmental toxicity.

RESULTS

NOAEL and LOAEL:NOAEL (maternal) = 10 mg/kg/day

LOAEL (maternal) = 30 mg/kg/day based on clinical signs and decreased body weight gain

NOAEL (developmental) = 100 mg/kg/day

LOAEL (developmental) > 100 mg/kg/day

Rem arks: There was no mortality observed at any dose level. At 100 mg/kg/day, one dam exhibited

dehydration, unkempt appearance, loose feces and perioral wetness. At 30 mg/kg/day, one dam was

noted with perioral wetness, gasping, loose feces and urine strai ns. Decreased body weight gain (12

- 13%) was observed in maternal animals at 30 and 100 mg/kg/day during gestation days 6

- 15. Food

consumption was not consistently affected by treatment. There were no treatment

- related increases in the

incidence of fetal external, visceral, or skeletal abnormalities at any dose level.

CONCLUSIONS

Based on the results of this study, the Maternal NOAEL is 10 mg/kg/day, and the Maternal LOAEL is 30

mg/kg/day, based on clinical signs and decreased body weight gain. The D evelopmental toxicity NOAEL

is 100 mg/kg/day, and the Developmental toxicity LOAEL > 100 mg/kg/day. There was no evidence for

developmental toxicity of ADBAC in this study.

RELIABILITY CODE

2

REFERENCES

42351501 (MRID) Neeper

- Bradley, T. (1992) Devel opmental Toxicity Evaluation II of Alkyl Dimethyl

Benzyl Ammonium Chloride (ADBAC) Administered by Gavage to CD Rats: Lab Project Number:

91N0031. Unpublished study prepared by Union Carbide Chemicals and Plastics Co., Inc. Bushy Run

Research Center. 28 1 p.

(EPA

- HQ

- OPP

- 2006

- 0339

- 0019[1])Title: Developmental Toxicity Evaluation of Alkyl Dimethyl Benzyl Ammonium Chloride (ADBAC)

Administered by Gavage to New Zealand White Rabbits: Lab Project Number: 91N0032

The study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: ADBAC, CAS No. 68424

- 85

- 1

Remarks: 81.09%

METHOD

Method/guideline followed: Not stated

GLP: Not stated

Year study was performed: Not stated

Species/strain: New Zealand White rabbits

Se x: female

Route of administration: gavage

Doses: 0, 1, 3 or 9 mg/kg/day

Exposure period: on gestation days 6 through 18

Frequency of treatment: daily

Control group and treatment: 0 mg/kg/day

Post

- observation period: Not stated

Statistical methods: Not stated

Method remarks: In a developmental toxicity study in rabbits, ADBAC (81.09%) was administered at

doses of 0, 1, 3 and 9 mg/kg/day to pregnant New Zealand White rabbits (16/dose) by oral gavage on

gestation days 6 through 18.

RESULTS

NOAEL a nd LOAEL:

NOAEL (maternal) = 3 mg/kg/day

LOAEL (maternal) = 9 mg/kg/day based on clinical signs of toxicity

NOAEL (developmental) = 9 mg/kg/dayLOAEL (developmental) > 9 mg/kg/day

Remarks: There was no mortality or abortions at any dose level. Hypoac tivity and labored breathing

were observed at 9 mg/kg/day in 2 of 15 rabbits. There were no effects on maternal body weight, food

consumption, cesarean section observations or necropsy observations. In offspring, there was no

evidence of developmental to xicity at any dose level tested.

CONCLUSIONS

The maternal NOAEL is 3 mg/kg/day, and the Developmental NOAEL is 9 mg/kg/day. The Maternal

LOAEL is 9 mg/kg/day, based on clinical signs of toxicity, and the Developmental LOAEL is > 9

mg/kg/day. There wa s no evidence of developmental toxicity of ADBAC in this study.

RELIABILITY CODE

2

REFERENCES

42392801 (MRID) Neeper

- Bradley, T.; Kubena, M. (1992) Developmental Toxicity Evaluation of Alkyl

Dimethyl Benzyl Ammonium Chloride (ADBAC) Administered by Gav age to New Zealand White

Rabbits: Lab Project Number: 91N0032. Unpublished study prepared by Union Carbide. 179 p.

(EPA

- HQ

- OPP

- 2006

- 0339

- 0019[1])Title: Developmental Toxicity Dose Range

- Finding Study of ADBAC Administered by Gavage to New

Zealand White Rabbits: Lab Project Number: 54

- 603

The study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: ADBAC, CAS No. 68424

- 85

- 1

Remarks: 81.09% pure

METHOD

Method/guideline followed: 870.3700

GLP: Not stat ed

Year study was performed: Not stated

Species/strain: New Zealand White rabbits

Sex: female

Route of administration: gavage

Doses: 0, 1, 3, 10, 30 or 60 mg/kg/day

Exposure period: gestation days 6 through 18

Frequency of treatment: daily

Control group and treatment: 0 mg/kg/day

Post

- observation period: Not stated

Statistical methods: Not stated

Method remarks: In a dose range

- finding study for developmental toxicity in rabbits, ADBAC (81.09%)

was administered at doses of 0, 1, 3, 10, 30 or 6 0 mg/kg/day to pregnant New Zealand White rabbits

(5/dose) by oral gavage on gestation days 6 through 18.

RESULTS

NOAEL and LOAEL:

NOAEL (maternal) = 3 mg/kg/day

LOAEL (maternal) = 10 mg/kg/day

, based on clinical signs and reduced body weight gain and food

consumption

LOAEL (developmental) not established.Remarks: Mortality was observed at doses of 30 and 60 mg/kg/day (2 and 5 doses, respectively).

Audible respiration was observed at doses greater than or equal to 10 mg/kg/day. At doses greater tha n or

equal to 30 mg/kg/day, clinical signs included hypoactivity, perioral wetness, and labored breathing. At

60 mg/kg/day, clinical signs included paralysis, cold extremities, prostration, slow respiration,

emaciation, loose feces, and perioral encrustat ion. Decreased body weight gain and food consumption

were observed at doses greater than or equal to 10 mg/kg/day. Developmental toxicity was not observed

at any of the doses tested.

CONCLUSIONS

The maternal NOAEL for the range

- finding study is 3 mg /kg/day and the LOAEL is 10 mg/kg/day, based

on clinical signs and reduced body weight gain and food consumption. There was no evidence of

developmental toxicity of ADBAC in this study.

RELIABILITY CODE

2

REFERENCES

42734401 (MRID) Chun, J.; Neeper

- Br adley, T. (1993) Developmental Toxicity Dose Range

- Finding

Study of ADBAC Administered by Gavage to New Zealand White Rabbits: Lab Project Number: 54

- 603.

Unpublished study prepared by Union Carbide. 118 p.

(EPA

- HQ

- OPP

- 2006

- 0339

- 0019[1])Title: Deve lopmental Toxicity Dose Range

- Finding Study of Alkyl Dimethyl Benzyl Ammonium

Chloride (ADBAC) Administered by Gavage to CD Rats: Lab Project Number: 54

- 613

The study is: Adequate Not Adequate

TEST SUBSTANCE

Identit y: ADBAC, CAS No. 68424

- 85

- 1

Remarks: 81.09% pure

METHOD

Method/guideline followed: 870.3700

GLP: Not stated

Year study was performed: Not stated

Species/strain: CD rats

Sex: female

Route of administration: gavage

Doses: 0, 25, 50, 100, 200 or 400 mg/kg/day

Exposure period: gestation days 6 through 15

Frequency of treatment: daily

Control group and treatment: 0 mg/kg/day

Post

- observation period: Not stated

Statistical methods: Not stated

Method remarks: In a dose range

- finding study fo r developmental toxicity in rats, ADBAC (81.09%)

was administered at doses of 0, 25, 50, 100, 200 or 400 mg/kg/day to CD rats (5/dose) by oral gavage on

gestation days 6 through 15, inclusive.

RESULTS

NOAEL and LOAEL:

NOAEL (maternal) = 50 mg/kg/day

LOAEL (maternal) = 100 mg/kg/day , based on significantly increased incidence of perioral wetness

NOAEL (developmental) = 100 mg/kg/dayLOAEL (developmental) = 200 mg/kg/day

, based on no survival

Remarks: Doses ≥ 200 mg/kg/day resulted in 100% mortality; necropsy findings revealed a distended

and change in color of the stomach, and distended intestines filled with mucoid fluid. These dams also

exhibited clinical signs including loose feces, perioral wetness and perioral encrustation, ataxia,

hypoactivity, urogenital area wetness and audible respiration. Maternal toxicity observed at 100

mg/kg/day was manifest as significantly increased incidence of perioral wetness.

CONCLUSIONS

The maternal NOAEL and LOAEL are 50 and 100 mg/kg/day, respectively. The NOAEL for

developmental toxicity is 100 mg/kg based on no survival of dams at 200 and 400 mg/kg/day.

RELIABILITY CODE

2

REFERENCES

42645101 (MRID) Chun, J.; Fisher, L. (1993) Developmental Toxicity Dose Range

- Finding Study of

Alkyl Dimethyl Benzyl Amm onium Chloride (ADBAC) Administered by Gavage to CD Rats: Lab

Project Number: 54

- 613. Unpublished study prepared by Bushy Run Research Center (BRRC). 104 p.

(EPA

- HQ

- OPP

- 2006

- 0339

- 0019[1])

### PHARMACOKINETIC STUDIES

Title: Absorption, Distribution , Metabolism and Excretion Studies of Alkyl Dimethyl Benzyl

Ammonium Chloride (ADBAC) in the Rat

TEST SUBSTANCE

Identity: ADBAC, CAS No. 68424

- 85

- 1

Remarks: purity 30%

METHOD

Method/guideline followed: 870.7485

Test type: in vivo

Species/strain/c ell type or line: Sprague

- Dawley rats

Sex: Both

Age and body weight range of animals used: Not stated

Number of animals/sex/dose: not stated

Route of administration: Oral or intravenous

Vehicle: Not stated

Doses:

Group 1: single dose 10 mg/kg (oral or intravenous)

Group 2: 100 ppm ADBAC in diet for 14 days

Group 3: single oral dose of 50 mg/kg

Excretion routes, body fluids, and tissues monitored and/or sampled during study: urine, feces,

unspecified tissues

Statistical methods used: Not stat ed

Method remarks: Disposition of ADBAC was examined in male and female Sprague

- Dawley rats

following a 10 mg/kg single dose by the oral or intravenous route, following exposure to 100 ppm

ADBAC for 14 days in the diet, or after a single oral dose of 50 mg/kg. Ring

- labeled test material was

used.

RESULTS

Detailed results:

Following oral administration, from 5

- 8% of the administered dose was eliminated in urine and 90

- 98%in feces. No apparent differences in disposition were noted between sexes. Fol lowing intravenous

administration, males eliminated 31% of the dose in urine and 44% in feces, while females eliminated

21% in urine and 55% in feces following intravenous administration. After oral administration, from

0.03%

- 0.58% of the administered d ose was accounted for in tissues. After intravenous administration,

tissue residues accounted for 33

- 36% of the dose and were observed mainly in the carcass. The results of

this study indicate that a majority of an administered dose of ADBAC is eliminate d in feces and involves

biliary excretion.

Metabolites measured: Not stated

CONCLUSIONS

The results of this study indicate that a majority of an administered dose of ADBAC is eliminated in feces

and involves biliary excretion.

REFERENCE S

40990701 (M RID) Selim, S. (1 989) Absorption, Distribution, Metabolism and Excretion Studies of

Alkyl Dimethyl Benzyl Ammonium Chloride (ADBAC) in the Rat: BTC Study No. P01 359.

Unpublished study prepared by Biological Test Center. 247 p.

Supplements:

41087701 (MR ID) Lin, P.; Selim, S. (1989) Addendum to Report Entitled "Absorption,

Distribution, Metabolism and Excretion Studies of Alkyl Dimethyl Benzyl Ammonium Chloride

(ADBAC) in the Rat" MRID 40990701 : Study No. PO1 359. Unpublished study prepared by Biological

Test Center. 44 p.

and

44783401 (MRID) Mazur, P. (1 999) Absorption, Distribution, Metabolism and Excretion

(ADME) Studies of Alkyl Dimethyl Benzyl Ammonium Chloride (ADBAC) in the Rat and

Addendum: Lab Project Number: P01359. Unpublished study prepared by ADBAC Quat Joint

VentureIChemical Specialities Manufacturers Assoc. 4 p.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 Aquatic Plants Yes Yes No

Health Effects

Acute Toxicity Yes Yes No

Repeated

- Dose Toxicity Yes Yes No

Genetic Mutation Yes Yes No

Chromosomal Aberrations Yes Yes No

Toxicity to Reproduction Yes Yes No

Developmental Toxicity Yes Yes No