**UNITED STATES ENVIRONMENTAL PROTECTION AGENCY WASHINGTON, D.C. 20460** 

**OF FICE OF CHEMICAL SAFETY**  **AND POLLUTION PREVENTION July 29**  **, 2014 MEMORANDUM** 
**SUBJECT:** Petition IN
- 10631; C.I . Pigment Red 112 ; Human Health Risk Assessment and Ecological Effects Assessment to Support Proposed Exemption from the Requirement of a Tolerance When Used as In ert Ingredients in Pesticide Formulations
. **All PC Code:** 804201 **DP Barcode:** 009297298 **Regulatory Action:** Inert Tolerance Exemption; 40 CFR 180. 920 **Petition No:**  IN
- 10631 **All CAS No :**  6535
- 46
- 2

**FROM:** Deirdr e Sunderland, Industrial Hygienist

In ert Ingredient Assessment Branch (IIAB) Registration Division (7505P)

**TO:** PV Shah , Chief

In ert Ingredient Assessment Branch (IIAB) Registration Division (7505P)2 Table of Contents Executive Summary ...........................................................................................3 1.0 Background................................................................................................5 2. 0 In ert Ingredient Profile....................................................................................5 2.1 Summary

of Uses.......................................................................................5 2.2 Phy sical and Chemical Properties.....................................................................5 2.3 Metabolism and Pharmacokinetics.......................................
. ............................6 3.0 Hazard Assessment.......................................................................................7 3.1 Acute Toxicity... ........................................................................................7 3.2 Subchronic Toxicity.....................................................................................8 3.3 Reproductive and Developmental Toxicity.........................................................9 3.4 Neurotoxicity ............................................................................................9 3.5 Mutagenicity
............................................................... ..............................9 3.6 Carcinogenic Potential................................................................................11 3.7 Endpoint Selection and Levels of Concern.........................................................11 3.8 FQPA Safet y Factor Considerations................................................................11 4.0 Exposure Assessment.................................................................. ..................12 4.1 Dietary

Exposure.......................................................................................12 4.2 Cancer Exposure........................................................................................12 4.3 Residential Exposure..................................................................................12 4.4 Occupational Exposure......................................................
. .........................12 5.0 Aggregate Risk...........................................................................................12 5.1 Acute Dietary

Risk....................................................................................13 5.2 Short
- term/Intermediate Aggregate Risk...........................................................13 6.0 Cumulative Risk................................................
.................................. .........13 7.0 Environmental Fate.......................................
. ...............................................13 8.0 Ecological Effects.................................................................................
. ......14 8.1 Invertebrate Toxicity
........................................................................... . .......14 8.2 Algal Toxicity
........................ . ............................................................ . ......143

**E XECUTIVE SUMMARY**  On September 5, 2013 , Exponent , on behalf of Clariant Corporation , submitted a petition to the Environmental Protection Agency , herein referred to as the agency or EPA, to amend 40 CFR 180. 920 by establishing an exemption from the requirement of a tolerance for the use of the inert ingredient C.I
. Pigment Red 112 ( CAS Reg. No. 6535
- 46
- 2) as a seed treatment pigment in p esticide formulations applied pre - harvest not to exceed 10% weight/weight ( w/w )
. These seeds will not be used for food, feed, or oil processing. C.I . Pigment Red 112, also known as 3
- h y dro x y- N
- ( 2
- m e th y

lphe n y l )
- 4
- [ 2
- (2,4, 5
- tr ichlo r oph e n y l ) diaz e n y l ]
- n a phthal e n e - 2
- carbo x am i de, is a sy nthetic az o compound p ri m ari l y used in industrial a ppl ic at i ons, such a s a c ol o ring a g e nt f o r p a int s, pl a st i c s and inks. C.I
. Pigment Red 112 has no approved pesticidal uses and the Agency is unaware of an y current residential. Based on the physical and chemical properties of C.I. Pigment Red 112 it is unlikely that C.I . Pigment Red 112 will be absorbed in the body and therefore, it is unl i k e l y that C
. I
. P i gm e nt R e d 112 b ec omes s ystemi ca l l y bioav a i l a

ble a ft e r exposure. In addition, azo reduction is likely to occur at low levels following oral ingestion limiting the potential for absorption and the subsequent formation of metabolites. It is expected that C.I . Pigment Red 112 will pass though the body and be excreted in the feces. The solubility

of C.I . Pigment Red 112 in octanol is 3.31 mg/L resulting in a calculated Log Kow of 2.5 which would indicated that the potential for bioaccumulation would be low. Acute studies revealed low oral and dermal toxicity (Toxicity Category IV). When tested on rabbits, the chemical was shown to be non
- irritating to the skin and eyes. Skin sensitization studies in guinea pigs showed that C.I. Pigment Red 112 was not skin sensitizer. In a 28
- day

oral toxicity

study in rats n o treatment - related changes were noted in neurological evaluations, body weights, food consumption, hematology or clinical chemistry

anal ys es, or organ weights at the limit dose of 1000 mg/kg/day
. Althou gh the r e w e r e no r ep r odu c t i ve o r d e v e lo p ment a l to x ici t y stud i e s av a i l a

ble for C
. I
. P i gment R e d 112, the 2 8
- d ay oral study in rats includ e d e ndpoint s specific to r ep rodu c t i ve tox i c i t y i n c lud i ng o r g an w e i ghts, gross p atho l o g y and his t op ath olo g y . N o adverse effect s were seen at doses up to 10 0 0 m g /k g /d a y . Inaddition , DEREK m o d e l i ng was conducted and did not ind i cate an y str u c tur a l a le rts f

or r ep rodu c t i ve to x ici t y or end o c rine
- r e l ated to x ici t y. No n e ur op atholo g i ca l c h an g e s o r e f fe c ts w er e rep o rt e d in any of the studies. The agency does not believe C.I. Pigment Red 112 will be neurotoxic For the purpose of this risk assessment three mutagenicity studies were reviewed: a reverse gene mutation study , an in vitro cell mutagenicity study, and a Chinese hamster cell in vitro study

. There was no evidence of cy

totoxicity or mutagenicity in an y of the reviewed studies. Although no carcinogenicity studies are available for C.I . Pigment Red 112 , C.I. Pigment Red 112 is not expected to be absorbed in the body based on the phy sical/chemical properties of the substance . Many azo dy es contain benzidine in their structure and this human bladder carcinogen may be released upon azo reduction of the dy e. In the case of C.I. Pigment Red 112 , azo reduction is likely to occur at low levels following oral ingestion limiting the potential for the formation of genotoxic and carcinogeneti c metabolites. Similarly , all4

mutagenicity

studies on C.I . Pigment Red 112 were negative which makes it unlikely that C.I . Pigment Red 112 would be carcinogenic. No endpoint of concern was identified for any of the acute studies conducted. In addition, n o endpoint of concern was determined in the 28
- day study up to the limit dose of 1000mg/kg/day (highest dose tested). Since no endpoint of concern was identified in acute and subchronic studies and because C.I . Pigment Red 112 is not expected to be absorbe d by the body , a quantitative risk assessment for C.I. Pigment Red 112 was not performed. As part of its qualitative assessment, the Agency did not use safety factors for assessing risk, and no additional safety factor is needed for assessing risk to infants and children. Furthermore, C.I . Pigment Red 112 will be used at a maximum of 10% w/w in seed treatments only. It is expected to be tightly bound to soil so the likelihood of consumers being exposed to C.I . Pigment Red 112 as an inert ingredient through foliar ingestion or water consumption is nominal. Seeds that have been chemically treated may NOT be used for food, feed, or oil processing
. The proposed use of C.I. Pigment Red 112 as a seed treatment/dye under 40 CFR 180 .920 is not expected to result in residential exposure to this chemical. In addition, there are no toxicological effects of concern in available studies and therefore, it is not necessary to conduct a quantitative assessments of residential (non
- occupation al) exposures and risks. Similarly , a quantitative occupational risk assessment is not necessary
. Based on ecological and fate data the agency concluded that C.I . Pigment Red 112 does not pose an ecological risk to terrestrial or aquatic species. C.I. Pigment Red 112 is a solid material with a low solubility in water and is not expected to volatilize. As a result of the low water solubility coupled with the projected partitioning of C.I . Pigment Red 112 , it is expected to be bound to soil and therefore, not expected to leach into ground water, be found in runoff or be taken up b y the plant. Although C.I. Pigment Red 112 may

persist in soils, due to the small amount of pigment used compared to the weight of the seeds used per acre, deposition in soil is not expected to significantly impact soil organisms. Bioaccumulation is also expected to be low. Based on the evaluated toxicity studies coupled with the expected exposure from the use of these chemicals as inert ingredients in pesticide products, EPA concludes that there is a reasonable certainty that no harm will result to the general population, including infants and children, from aggregate exposure to residues of C.I . Pigment Red 112 when used as an i nert ingredient in pesticide formulations for seed treatment at a maximum of 10% w/w under 40 CFR 180.920
.5

**1.0 B ACKGROUND**  On September 5, 2013, Exponent, on behalf of Clariant Corporation, submitted a petition to the EPA, to amend 40 CFR 180.920 by establishing an exemption from the requirement of a tolerance for the use of the inert ingredient C.I. Pigment Red 112, CAS Reg. No. 6535
- 46
- 2 for use as a seed treatment pigment in pesticide formulations , not to exceed 10% w/w, applied pre
- harvest. By regulation, seeds treated with a pesticide, with the exception of those used “solely as at - planting or hopper box treatments” or “products which are gaseous in form or are used as fumig ants”, must also be treated with a “colorant” to indicate that the seeds are, in fact, treated with a pesticide (40 CFR 153.155). According to the Federal Seed Act section 201.31a (d), seeds that have been chemicall y treated may not be used for food, feed , or oil processing. C.I
. Pigment Red 112, also known as 3
- h y d ro x y- N
- ( 2
- m e th y

lphe n y l )
- 4
- [ 2
- (2,4, 5
- tr ichlo r oph e n y l ) diaz e n y l ]
- n a phthal e n e - 2
- carbo x am i de, has no currently a pproved food or nonfood pesticidal uses. This chemical was th e subject of an IUC LID Chemical Safet y report prepared by C lariant In c. in 2010. To access the Office of Chemical Safet y and Pollution Prevention ( OCSPP ) test guidelines referenced in this document electronically , please go to http://www.epa.gov/ocspp and select “Test Methods and Guidelines.” **2.0 INERT INGREDIENT PROFILE**  **2.1 Summary of Uses**  C.I . Pigment Red 112 is p ri m ari l y used in industrial a ppl ic at i ons , such a s a c ol o ring a g e nt f o r p a int s, pl a st i c s and inks. C.I . Pigment Red 112 has no approved pesticidal uses. According to the U.S. National Institutes of Health ’s Household P roduct Database C.I. Pigment Red 112 is not found in any household consumer goods. It has been used in other countries in cosmetics, children’s finger and face paints, and permanent tattoo ink . ( Environment Canada, 2013 ) There are , however, no Federal Drug Administration (FDA) approved cosmetic uses for C.I . Pigment Red 112 in the US. **2.2 Physical**  **and Chemical Properties**  Some of the phy sical and chemical characteristics of C.I . Pigment Red 112 along with the chemical structure and nomenclature, are found in the table below.6

**Table 1. Physical and Chemical Properties of**  **C.I. Pigment Red 112**  **Parameter Value**  **Reference**

Structure ChemID Plus , 201 3

CAS Reg. No. 6535
- 46
- 2 ChemIDPlus, 201 3 Molecular Formula C 24 H 16 C 13 N 3 O 2 ChemIDPlus, 201 3 Molecular Weight 48 4 .76 Clariant , 2010 Other Names 3
- h y d ro x y- N
- ( 2
- m e th y

lphe n y l )
- 4
- [ 2
- (2,4, 5
- tr ichlo r oph e n y

l ) diaz e n y l ]
- n a phthal e n e - 2
- carbo x am i de ; CI12370 ChemIDPlus, 201 3 Physical State solid Clariant , 2010 Melting Point None, decomposition at ≥ 270°C Clariant , 2010 Particle Size distribution Medium:4.51 μm Clariant , 2010 Density 1.48 g/ml Clariant , 2010 Vapor Pressure 8.51 x10 - 11 mm Hg (calculated) Clariant , 2010 Log K ow @ 25°C 2.5 (calculated) Clariant , 2005, (MRID 49336101) Water Solubility 9.8μg/L Clariant , 2005, (MRID 49336101)
**2.3 Metabolism**  **and Pharmacokinetics** C.I . Pigment Red 112 has very low water solubility

( 9.8 μ g/ L) and therefore, it is unlikely that C.I
. Pigment Red 112 will be absorbed b y the bod y and become s ys temicall y bioavailable
. In addition, azo reduction is unlikely

to occur following oral ingestion limiting the potential for absorption and the subsequent formation of metabolites. It is expected that C.I . Pigment Red 112 will pass though the body and be excreted in the fec es. This is supported b y the 28
- day oral study which found reddish staining in the feces and gastrointestinal tract of rats treated with C.I . Pigment Red 112
. The solubility of C.I. Pigment Red 112 in octanol is 3.31 mg/Lresulting in a calculated Lo g Kow of 2.5 which would indicated that the potential for bioaccumulation would be low. Based on the physical and chemical properties of C.I . Pigment Red 112 it is unlikely that C.I . Pigment Red 112 will be absorbed in the body and therefore, it is unl i k e l y that C
. I
. P i gm e nt R e d 112 b ec omes s ystemi ca l l y bioav a i l a ble a ft e r exposure.7

**3.0 H**  **AZARD ASSESSMENT 3.1**  **Acute Toxicity** Acute Oral Toxicity (OSCPP 870.1100) Wistar rats (5/sex/dose) were administered 5000 mg/kg of C.I. Pigment Red 112 (97% C.I. Pigment Red 112 ) via g avage and observed for 14 day s. No mort alities occurred at 500 0 mg/kg
. ( MRID 49148201 ) **LD 50**  **> 5000 mg/kg**  According to the 2010 IUCLID report on C.I . Pigment Red 112 , various other acute oral (gavage) rat studies were conduct ed with LD 50 s ranging from >10,000 to > 15,000 mg/kg. These studies were not available to the Agency

and were reported in the IUCLID document as “reliable with restrictions” and therefore, they

are presented here as supporting studies only
. Acute Dermal Toxicity (OSCPP 870.1200) Albino Wistar rats (5 males ) were adm inistered a single dose of 5000 mg/kg of C.I . Pigment Red 112 applied directl y to the skin for 24 hours and were observed for 14 days after removal of excess test substance. No mortalit y or signs of toxicity were observed. (MRID 4914 8202 ) **LD 50**  **> 5000 mg/kg** Acute Inhalation Toxicity No acute inhalation toxicity studies are available for C.I . Pigment Red 112
. Primary Dermal Irritation (OSCPP 870.2500) A test substance comprised 97% C.I. Pigment Red 112 was tested for acute skin irritation when a single dermal dose of 500 mg applied to a 2.5 x 2.5 cm surgical gauze patch and applied to an clipped area of the skin of three albino New Zealand White rabbits held in place with semi
- occlusive for four hours (O SCPP 870.2500)
. Skin examinations were performed at 30 minutes, 24, 48, and 72 hours after removal of dressing. The skin areas treated were slightly stained light pink until 24 hours after removal of the patch; however, the evaluation of skin redness was s till possible despite the slight staining. At 30 minutes post exposure one animal display ed very slight edema and ery

thema. No reactions were observed in an y of the animals at 24 hours after the removal of the test substance. (MRID 49148203 ) **Based on the results of this study C.I. Pigment Red 112**  **is not considered a skin irritant**  **.** Primary Ey e Irritation (OSCPP 870.2400) A test for acute ocular irritation was conducted with C.I . Pigment Red 112 by appl yi ng a single dose of 100 mg (as a paste in 0.05 ml poly eth yl ene gl ycol ), into the conjunctival sac of three Albino New Zealand White rabbits (OSCPP 870.2400). One eye remained untreated and served as a control. Treated eyes were rinsed 24 hours after applicat ion. Ocular examinations were performed at 1, 24, 48, and 72 hours after instillation. No iritis or corneal opacity was observed throughout the observation period in any of the animals. Conjunctival redness was observed at 1 hour in all animals (score 1) , and in one animal 24 hours after instillation (score 1). Chemosis (score 1) was observed in one animal at 1 hour after instillation. All8

effects cleared b y 48 hours post instillation. **Based on the results of this study**  **C.I. Pigment Red 112 is considered**  **nonirritating.** (MRID 49148204) **Table 2**  **. Summa**  **r y**  **of Acute Toxicity Data**  **Endpoint DMS**  **Toxicity Category Reference**  Acute Oral LD 50 (mg/kg) >5000 IV MRID 49148201 Acute Dermal LD 50 (mg/kg) >5000 IV MRID 49148202 Acute Inhalation LD 50 (mg/L) No Data N/A N/A Skin Irritation (4 hour exposure) Not a skin irritant IV MRID 49148203 Eye Irritation Not an eye irritant IV MRID 49148204 Dermal Sensitization Skin sensitization testing of C.I . Pigment Red 112 (containing 3% Naphtol AS
- D) was performed on 20 female guinea pigs according to the Buehler method (OCSPP 870.2600). Due to the red color of the test substance a determination of ery thema could not be made using a test substance of 100% C.I. Pigment Red 112; therefore, the study port ion for ery

thma was performed with a 20% pigment composition in sesame oil with a control group (10 animals) receiving sesame oil only
. After 24 hours, no irritating effects (ery thema or edema) were observed following the application of concentrations of 100% or 20% C.I. Pigment Red 112. **Based on the results of this study, the mixture was not**  **considered to be a dermal sensitizer**
. ( MRID 49148206 ) Two L ocal Ly

mph Node Assay s were conducted with C.I . Pigment Red 112; one study tested 0.5, 10, and 20% C.I. Pigment Red 112 with <0.01% Naphtol AS
- D (MRID 49148205) and the other tested 5, 10, and 20% C.I. Pigment Red 112 with a suggested 4.1% Naphtol AS
- D (MRID 49148214). The second test substance containing what the petitioner claimed was 4.1% Naphtol AS
- D was positive for skin sen si tization whereas the test with 0.01% Naphtol AS
- D was not , indicating that skin sen si tization is possible when Naphtol AS
- D is present at sufficiently high concentrations. The percent of Napthol AS
- D in the second study could not be confirmed with the study data available. However, the two negative skin sensitization studies did have documented proof of the Napthol AS
- D content at up to 3%. The manufacturing process for C.I . Pigment Red 112 that will be used as a seed treatment inert ingredient is specifically

manufactured to contain less than 1% Naphtol AS
- D which , based on the study results , would not result in skin sensitization. Therefore, the C.I. Pigment Red 112 manufactured for use as a see d treatment inert ingredient is not considered a skin irritant. **3.2 Subchronic Toxicity**  In a 28
- day

oral toxicity

study (OCSPP 870.3050) C
. I
. Pigment Red 112 (99.15%) was administered in propylene gl ycol to 5 Crl:WI (Han) rats/sex/dose by gavage at dose levels of 0, 100, 300, or 1000 mg/kg/day

dail y for 28 days. Animals were observed daily for clinical signs; body

weight and food consumption were recorded weekl y; and selected neurological evaluations ( i.e., hearing ability , pupillary reflex, stat ic righting reflex, grip strength, and motor activity testing (recording period of 12 hours overnight using a computerized monitoring s ys tem) ) were performed at the end of Week 1, 2, 3, and 4.9

Gross and microscopic examination (e.g., hematology and clinica l chemistry , tissue and organ examination, and organ weights) was conducted at study termination after 4 weeks of treatment. Treatment
- related effects were limited to red staining of feces in all treated males and females and staining of bod y surfaces, b ut this pigmentation was not apparent in the slides of tissues examined microscopicall y and was not considered adverse. No treatment
- related changes were noted i n neurological evaluations, body weights, food consumption, hematology or clinical chemistry

an al ys es, or organ weights. Although a LOAEL was not established, the limit dose was tested. (MRID 49148210) **The NOAEL was 1000 mg/kg/day, and the LOAEL was not established. 3.3**  **Reproductive and Developmental**  **Toxicity** Althou gh the r e w e r e no r ep r odu c t i ve o r d e v e lo p ment a l to x ici t y stud i e s av a i l a ble for C
. I
. P i gment R e d 112, the 2 8
- d ay oral study in rats includ e d e ndpoint s specific to r ep rodu c t i ve tox i c i t y i n c lud i ng o r g an w e i ghts, gross p atho l o g y and his t op ath olo g y . The stu d y showed that the r e w e re no tr eatm e nt - r e lat e d e f f e c ts o n r e lative and a

bsolute o rg an w e i ght s f

o r the test e s or epid i d y

m i d e s . G ross p atho l o g y of th e r ep r odu c t i ve o r g ans did not r ev ea l an y tr eatm e nt - r e lat e d e f f ec ts. F urthe r

m o r e , his t op atho l o g y o f epid i d y m i d e s, p r ostate g land, s e m i n a l v e sicl e s, test e s, ov ari e s, v a g ina, c e rvi x , and ute r us demonstrated no effect at doses up to 10 0 0 m g /k g /d a y . Inaddition to this study DEREK m o d e l i ng was conducted and did not ind i cate an y str

u c tur a l a le rts f

or r ep rodu c t i ve to x ici t y or end o c rine
- r e l ated to x ici t y. **3.4 Neurotoxicity**  No adverse effects on selected neurological evaluations (i.e., hearing ability, pupillary reflex, static righting reflex, grip strength, and motor activity testing ) were observed in the 28
- day toxicity study in rats. **3.5 Mutagenicity** **/Cytotoxicity** Reverse Gene Mutation (MRID 49148207) In a reverse gene mutation assay (OCSPP 870.5100) in bacteria, S. typhimurium strains TA98, TA100, TA1535, and TA1537 were exposed to C.I. Pigment Red 112 dissolved in Dimethyl sulfoxide (DMSO) in two independent assays using triplicate plating. Experiment 1, which was both a mutagenesis and a range - finding assay , used the standard plate assay and tested concentrations of 0, 4, 20, 100, 500, 2500, and 5000 μg/plate, with and without S9
- mix derived from livers of induced rats. In addition, it tested the same concentrations with metabolic activation in a preincubation assay that used S9
- mix derived from livers of non
- induced hamsters. The confirmatory mutagenicity assay (experiment 2) tested mutagenesis and cy totoxicity under exactly

the same three sets of conditions as in experiment 1. For the tests with metabolic activation, the S9 fraction for the standard plate assay was obtained from Aroclor
- 1254
- induced male Sprague
- Dawley rat liver and that for the preincubation test was obtained from non
induced, male, S yrian golden hamster liver. C.I . Pigment Red 112 was tested at concentrations up to the limit concentration of the assay in both experiments. There was no evidence that the assays were affected b y cy totoxicity . Precipitate was observed on all test plates at concentrations of 500 μg/plate and above. While the precipitate did not interfere with the counting of the numbers of revertant colonies, it did make it impossible to evaluate10

bacteri al lawns at the maximum concentration tested. The number of revertants per plate was not increased over the concurrent solvent control value at an y test material concentration, with or without S9
- mix, in any

tester strain. The solvent and positive controls induced the appropriate responses in the corresponding strains. **There was no evidence of induced mutant colonies over background.** In vitro Cell Mutagenicity (MRID 49148209) In an in vitro mammalian cell gene mutation assay (OSCPP 870.5300) , Chinese hamster V79 cells cultured in vitro were exposed to C
. I
. Pigment Red 112 dissolved in DMSO and culture medium for 4 hours at concentrations of 0, 3.1, 6.3, 12.5, 25.0, 50.0, and 400 μg/mL in the absence or the presence of metabolic activation in the first experiment or for 24 hours at concentrations of 0, 3.1, 6.3, 12.5, 25.0, 50.0, and 400 μg/mL in the absence of metabolic activation in the second experiment. All doses except for 50.0 μg/mL were continued to determine mutant frequencies. The S9 f raction was obtained from phenobarbital and β
- naphthoflavone
- induced male Wistar Hanlbm rat liver. C.I . Pigment Red 112 was tested up to the limit of solubility in DMSO and aqueous media. In both the presence and absence of metabolic activation, the test article was tested up to 16 times the lowest concentration at which precipitation was consistently observed with the naked ey e in the culture medium. There was no convincing evidence of any cy totoxicity . No significant increase in mutant frequency over that of the solvent controls was observed for any test article concentration. The positive controls did induce the appropriate responses. **There was no evidence of induced mutant colonies over background.**  Chinese Hamster Cell in vitro (MRID 49148208) In an in vitro test on the induction of micronuclei in cultured V79 Chinese hamster cells, V79 cell cultures were treated with Pigment Red 112 dissolved in DMSO, for 4 or 24 hr without activation, and for 4 hr with activation. The preparation interval was 24 h in all tests. In order to assess the toxicity of the test article to the cultured V7 cells, a cell Proliferative Index (PI) was calculated for all treated and control V79 cultures. Only those cultures whi ch showed a PI > 1.3 were evaluated. Con centrations of the test article selected for micronucleus anal ys is were 7.8, 15.6 and 31.3 P μg/mL(Test 1, without activation), 3.9, 7.8 and 15.6 P μg/mL(Test 2, without activation), 2.0, 3.9 and 7.8 μg/mL(Test 1, w ith activation), and 2.0, 3.9 and 7.8 μg/mL(Test 2, with activation). With activation, the maximum concentrations tested for micronucleus induction were well below the limit of solubility (7.8 μg/mL was the highest concentration used in either Test with a ctivation, while 31.3 μg/mLwas the concentration at which precipitation was first observed). S9 came from male Wistar rat liver induced with phenobarbital and β
- naphthoflavone. **The test article was found to cause no significant increases in micronucleus induction in cultured**  **V79 cells without or with activation at the concentrations that were tested** . However, the highest concentration tested with activation was well below a precipitating concentration, and no data were presented to indicate that higher co ncentrations were cy totoxic. P =Precipitating Concentration Supporting Data Although the full stud y reports were not available, the IUCLID Chemical Safety Report for C.I. Pigment Red 112 illustrates two additional Bacterial Reverse Mutation Assays which support the studies11

presented above. In both studies no evidence of mutagenicity or cy totoxicity was seen with and without metabolic activation in S. ty phimurium TA 1535, 1537. 98, or 100 or E.coli WP uvr A
. **3.6 Carcinogenic Potential**  No carcinogenicit y studies are avai lable for C.I. Pigment Red 112 . In the absence of quantitative data a qualitative structure activity relationship (SAR) database, DEREK Version 11, was used to determine if there were structural alerts suggestive of carcinogenicity . A lthough DEREK results indicate “plausible” alerts for carcinogenicit y , this is unlikely to be the case because C.I. Pigment Red 112 is not expected to be absorbed in the bod y based on the large molecular weight and low water solubility . Man y azo dyes contain benzidine in their structure and this human bladder carcinogen may be released upon azo reduction of the dye . (Levine, 1991) In the case of C.I . Pigment Red 112 , azo reduction i s likely to occur at low levels following oral ingestion limiting the potential for the formation of genotoxic and carcinogenetic metabolites. ( Environment Canada , 2013 ) Similarly , all mutagenicit y studies on C.I
. Pigment Red 112 were negative which makes it unlikely that C.I. Pigment Red 112 would be carcinogenic. **3.7 Endpoint Selection and Levels of Concern**  Available toxicity studies indicate that C.I . Pigment Red 112 has a very low acute and subchronic toxicity
. No endpoint of concern was identified for any of the acute studies conducted. In addition, no endpoint of concern was determined in the 28
- day study up to the limit dose of 1000mg/kg/day (highest dose tested). S ince no endpoint of concern was identified in acute and sub chronic studies and because C.I . Pigment Red 112 is not expected to be absorbed by the body based on th e phy sical/chemical properties, a quantitative ri sk assessment for C.I . Pigment Red 112 is n ot necessary. **3.8 FQ**  **PA Safety Factor C**  **onsiderations** FFDCA Section 408(b)( 2)(c) provides that EPA shall apply an additional tenfold (10X) margin of safety for infants and children in the case of threshold effects to account for prenatal and postnatal toxicity and the completeness of the database on toxicity and exposure unless E PA determines based on reliable data that a different margin of safety will be safe for infants and children. This additional margin of safety is commonly referred to as the FQPA safety factor (SF). In applying this provision, EPA either retains the defaul t value of 10X, or uses a different additional safety factor when reliable data available to EPA support the choice of a different factor. The available acute and repeat dose toxicity studies indicate that C.I . Pigment Red 112 has low toxicity with no ef fects seen at the limit dose of 1000 mg/kg/day. DEREK m o d e l i ng was conducted and did not ind i cate an y str u c tur a l a le rts f

or r ep rodu c t i ve to x ici t y or e nd o c rine
- r e l ated to x ici t y. There is no indication, based upon the available data, that C.I . Pigment Red 112 is neurotoxic or immunotoxic and therefore, would not result in increased susceptibility in infants or children. Although the toxicity database for C.I . Pigment Red 112 is limited to acute studies, a 28
- day repeat dose study and mutagenicity studies th e phy sical chemical properties suggest that it is unlikely that C.I . Pigment Red 112 will be absorbed by the body and become bioavail able and therefore , unlikely to cause adverse effects in infant and children. As part of its qualitative assessment, the Ag ency

did not use safety

factors for assessing risk, and no additional safety factor is needed for assessing risk to infants and children. Furthermore , C.I . Pigment Red 112 will be used at a maximum of 10% w/w in seed treatments onl y. It is expected to be t ightly bound to soil so the likelihood of consumers12

being exposed to C.I . Pigment Red 112 as an inert ingredient through foliar ingestion or water consumption is nominal
. T aking int o c onsid e r at i on a ll av a i l a ble info r mation , there is no concern, at this time, for increased sensitivity to infants and children to this chemical when used as inert ingredient in pesticides formulations. **4.0 EXPOSURE**  **4.1 Dietary Exposure**  Although dietary

exposure to C.I . Pigment Red 112 may occur from eating foods grown from seeds treated with pesticide formulations containing this inert ingredient and drinking water containing runoff from soils containing the treated seeds , the amount of residue that a consumer would be exposed to would be relativel y small because the C.I. Pigment Red 112 is expected to be tightl y bound to soil and therefore , it is unlikely

the pigment would be taken up by the plant and subsequently available for consumption through eating the plant. In addition, runoff is unlikely based on the low water solubility and soil binding of the pigment. T he amount of C.I. Pigment Red 112 used in formulation is not to exceed 10% w/w. According to the Federal Seed Act section 201.31a (d), seeds that have been chemicall y treated m ay

NOT be used for food, feed, or oil processing. There are no other known food uses for C.I. Pigment Red 112 . Furthermore , no endpoint of concern was identified and therefore, a quantitative dietary

exposure assessment for C.I. Pigment Red 112 was not con ducted. **4.2 Cancer E**  **xposure** Although there are no animal carcinogenicity studies available in the database, C.I . Pigment Red 112 tested negative in mutagenicity assay s and no evidence of specific target organ toxicity was observed in the repeat dose studies. Based on the available toxicity data, C.I . Pigment Red 112 not expected to be carcinogenic. Therefore, a cancer dietary exposure assessment is not necessary to assess cancer risk. **4.3 Residential (Non**  **- Occupational) Exposure**  The term “residential exposure” is used in this document to refer to non
- occupational, non
- dietary exposure (e.g., for lawn and garden pest control, indoor pest control, termiticides, and flea and tick control on pets). The proposed use of C.I. Pigment Red 112 as a seed treatment/dye under 40 CFR 180.920 is not expected to result in residential exposure to this chemical . In addition, there are no toxicological eff

ects of concern in available studies and therefore, it is not necessary to conduct a quantitati ve assessments of residential (non
- occupational) exposures and risks. **4.4 Occupational Exposure** Ty pically , seeds used in the propagation of agricultural crops are treated with one or more pesticide products. By

regulation, seeds treated with a pesticide, with the exception of those used “solel y as at planting or hopper box treatments” or “products wh ich are gaseous in form or are used as fumigants”, must also be treated with a “colorant” to indicate that the seeds are, in fact, treated with a pesticide (40 CFR 153.155). This is generall y done in a closed system with minimal worker exposure. According to the Federal Seed Act section 201.31a (d), seeds that have been chemically treated may NOT be used for food, feed, or oil processing. The Agency

has reviewed the available toxicological information for C.I. Pigment Red 112 and there are no adverse tox icological effects observed in the studies submitted at the limit dose of 1000 mg/kg/day (highest dose tested), therefore a quantitative occupational risk assessment is not necessary.13

**5.0 AGGREGATE RISK**  **5.1 Acute Aggregate Risk**  There was no hazard attributable to a single exposure seen the in the toxicity database for the C.I. Pigment Red 112 ; therefore an acute aggregate risk assessment is not required. **5.2 Short**  **- term/Intermediate Aggregate R** **isk** Short
- term aggregate exposure takes into ac count short
- term residential exposure plus chronic exposure to food and water (c onsidered to be a background exposure level). S ince there are no oral, dermal or inhalation toxicological endpoints of c oncern , it is not necessary to aggregate dermal and inha lation residential exposures with estimated dietary exposures. **6.0 CUMMULATIVE EXPOSURE**  Section 408(b)(2)(D)(v) of FFDCA requires that, when considering whether to establish, modify , or revoke a tolerance, the Agency consider “available information” concerning the cumulative effects of a particular pesticide's residues and “other substances that have a common mechanism of toxicity .” E PA has not found C.I . Pigment Red 112 to share a common mechanism of toxicity with an y other substances, and C.I. Pigment Red 112 does do not appear to produce a toxic metabolite produced b y other substances. For the purposes of this tolerance action, therefore, EPA has assumed that the C.I. Pigment Red 112 does not have a common mechanism of toxicity with oth er substances. For information regarding EPA's efforts to determine which chemicals have a common mechanism of toxicity and to evaluate the cumulative effects of such chemicals, see EPA's website at http://www.epa.gov/pesticides/cumulative
. **7 .0**  **ENVI RONMENTAL FATE**  In determining the environmental fate of C.I. Pigment Red 112 the Agency considered phy sical/chemical properties (Table 1) and modeled data from the EPISuite v4.11 the PBT Profiler ( Persistent, Bioaccumulative, and Toxic Profiles Estimated for Organic Chemicals )
. C.I . Pigment Red 112 is a solid material with a l ow solubility in water (9.8μ g/ L) and is not expected to volatilize (Vapor Pressure 8.51 x10 - 11 mm Hg ; Henry ’s Law Constant estimated 1.082E
- 12 atm
m 3 /mole ) . B ased on the results of the Level III Fugacity Model ( Table 3 ) C.I . Pigment Red 112 ’s highest affinity would be for sediment ( 52%) and soil (47%) . As a result of the low water solubility coupled with the projected partitioning of C.I. Pigment Red 112 , C.I . Pigment Red 112 is expected to be bound to soil and therefore, not expected to leach into ground water, be found in runoff or be taken up b y the plant
. Although C.I. Pigment Red 112 may persist in soil ( EPI Suite and PB T Profiler Estimate) , d ue to the small amount of pigment used compared to the weight of the seeds used per acre , deposition in soil is not expected to significantly impact soil organisms. Bioaccumulation is also expected to be low (Log Kow 2.5).14

**8.0 ECOLOGICAL EFFECTS**  **8 .**  **1 I**  **nv e**  **r t**  **eb r**  **a te**  **T o**  **x i**  **c i**  **t y**  **Daphnia magna Reproduction Test** A 21
- day Reproduction Test (OECD 211) was performed with Daphnia magna (ten/dose) under semi
static conditions. R el ate d to the total number of produced juveniles (dead + alive) t he percentage o f dead juveniles was <1 % , which is seen to be not biologicall y significant. Therefore, a nominal loading of 1 mg/L showed no biologically or statistically significant effects including mortality , immobilization, and reproduction. The No Observed Effect Loading (NOEL) and the Lowest Observed Effect Lo ading (LOEL) were assessed at 1 mg/L and > 1 mg/L , respectively. ( MRID 49148212 ) **Earthworm Reproduction Test -**  **Eisenia fetida** An 8
- week Earthworm Reproduction Test (OECD 222) was carried out under static conditions to determine the lethal and sublethal effects (i.e., mortality , biomas s and reproduc ti v e performance) o f C.I . Pigment Red 112 on the earthworm species Eisenia fetida (10/replicates(8)/dose) treated with a limit test item concentration of 1000 mg/kg dry soil. Adult earthworms were placed on art i ficia l s oil treated with the limit test item concentration as well as on artificial soil serving as contr o l and on control soil which was treated with the solvent. Application of the test item was carried out p r i or to i nt r oducing the earthw o rms into the test vessel s . After the first 4 weeks of the test, mortalit y and biomass o f the adult worms were determine d
. After a further 4 weeks the reproduction rate was determined by counting the hatched juvenile s . Under the conditions of this study C.I. Pigment Red 112 did not induce mortality or pathological symptoms of adult earthworms after exposure to the test item at a concentration of 1000 mg CI /kg soil dry weight for 28 day s. The biomass increase of the adult earthworms after 28
- days was not s tatistically si gnificantl y reduced compared to the pooled control at the limit test item concentration of 1000 mg/kg s o il dry weight. After 8
- weeks of exposure earthworm reproduction was not reduced, but instead increased at the limit test concentration of 1 000 mg CI /kg soil dry weight. Thus, the No Observed Effect Concentration (NOEC) with respect to mortality , body weight and reproduction of Eisenia fetida after 8 weeks of exposure in artificial so i l was determined to be at 1000 mg/kg soil dry weight (highest dose tested) and the Lo west Observed Effect Concentration ( LOEC ) was > 1000 mg/kg soil dry weight. ( MRID 49148213 ) **Table 3: Level III Fugacity Model Media**  **Mass Amount (%) Half**  **- Lif**  **e (days)** Air 0.026 0.41 Water 0.943 180 Soil 47 360 Sediment 52 162015

**8 .2**  **A l**  **g al**  **T o**  **x i**  **c i**  **t y**  **Alga Growth Inhibition Test -**  **Desmodesmus subspicatus** A 72
- hour Alga Growth Inhibition Test (OECD 201) was conducted under static conditions with unicellular freshwater green alga species Desmodesmus subspicatus to determine the effect of C.I. Pigment Red 112 on growth rate and yi eld. Six replicated were tested with the limit concentration (1mg/L) and the control. The NOEC was 1 mg/L. The LOEC and EC 50 were >1 mg/L . (MRID 49148211)
**9 .0**  **RISK CHARACTERIZATION** The Agency

is unaware of any

current residential or pesticidal uses for C.I. Pigment Red 112
. C.I. Pigment Red 112 is p ri m ari l y used in industrial a ppl ic at i ons, such a s a c ol o ring a g e nt f o r p a int s, pl a st i c s and inks. Under the proposed use pattern C.I . Pigment Red 112 will be uses in seed treatment at no more than 10% w/w to identify seeds that have been treated with a pesticide. These seeds will not be used for food, feed, or oil processing. Based on the physical and chemical properties of C.I. Pigment Red 112 it is unlikely that C.I . Pigment Red 112 will be absorbed in the body and therefore, it is unl i k e l y that C
. I
. P i gm e nt R e d 112 b ec omes s ystemi ca l l y bioav a i l a

ble a ft e r exposure . In addition, azo reduction is likely to occur at low levels following oral ingestion limiting the potential for absorption and the subsequent formation of metabolites. It is expected that C.I . Pigment Red 112 will pass though the body and be excreted in the f eces. The solubility

of C.I . Pigment Red 112 in octanol is 3.31 mg/L resulting in a calculated Log Kow of 2.5 which would indicated that the potential for bioaccumulation would be low. Acute studies revealed low oral and dermal toxicity (Toxicity Category IV). When tested on rabbits, the chemical was shown to be non
- irritating to the skin and eyes. Skin sensitization studies in guinea pigs showed that C.I. Pigment Red 112 was not skin sensitizer. In a 28
- day

oral toxicity

study Crl:WI (H an) rats were gavaged with 0, 100, 300, or 1000 mg/kg/day for 28 day s. Animals were observed dail y for clinical signs; body weight and food consumption were recorded weekl y; and selected neurological evaluations (i.e., hearing ability, pupillary reflex, st atic righting reflex, grip strength, and motor activity testing) were performed at the end of Week 1, 2, 3, and 4. Gross and microscopic examination s were conducted at study termination. Treatment
- related effects were limited to red staining of feces in al l treated males and females and staining of body surfaces, but this pigmentation was not apparent in the slides of tissues examined microscopicall y and was not considered adverse. No treatment
- related changes were noted in neurological evaluations, body we ights, food consumption, hematology or clinical chemistry

anal ys es, or organ weights. The NOAEL was 1000 mg/kg/day

(highest dose tested), and a LOAEL was not established. Althou gh the r e w e r e no r ep r odu c t i ve o r d e v e lo p ment a l to x ici t y stud i e s av a i l a ble for C
. I
. P i gment R e d 112, the 2 8
- d ay oral study in rats includ e d e ndpoint s specific to r ep rodu c t i ve tox i c i t y i n c lud i ng o r g an w e i ghts, gross p atho l o g y and his t op ath olo g y . The stu d y showed that the r e w e re no tr eatm e nt - r e lat e d e f f e c ts o n r e lative and a

bsolute o rg an w e i ghts f

o r the test e s or epid i d y

m i d e s. G ross p atho l o g y of th e r ep r odu c t i ve o r g ans did not r ev ea l an y tr eatm e nt - r e lat e d e f f ec ts. F urthe r

m o r e , his t op atho l o g y o f epid i d y m i d e s, p r ostate g land, s e m i n a l v e sicl e s, test e s, ov ari e s, v a g ina, c e rvi x , and ute r us demonstrated16

no effect at doses up to 10 0 0 m g /k g /d a y . Inaddition to this study DEREK m o d e l i ng was conducted and did not ind i cate an y str

u c tur a l a le rts f

or r ep rodu c t i ve to x ici t y or end o c rine
- r e l ated to x ici t y. Three mutagenicity

studies were reviewed in this risk assessment: a reverse gene mutation study , an in vitro cell mutagenicity

study , and a Chinese hamster cell in vitro study

. In the reverse gene mutation assay S. typhimurium strains TA98, TA100, TA1535, and TA1537 were exposed to C.I . Pigment Red 112 dissolved in Dimethyl sulfoxide (DMSO) in two independent assay s using triplicate plating. Experiment 1 used the standard plate assay with and without S9 and a preincubation assay up using S9 up to 5000 μg/plate. The confirmatory mutagenicity

assay (experiment 2) tested mutagenesis and cy totoxicity under exactly

the same three sets of conditions as in experiment 1. C.I . Pigment Red 112 was tested at concentrations up to the limit concentration of the assay in both experiments. There was no evidence that the assay s were affected b y cy totoxicity and no evidence of induced mutant colonies over background. In an in vitro mammalian cell gene mutation assay Chinese hamster V79 cells cul tured in vitro were exposed to C
. I
. Pigment Red 112 dissolved in DMSO and culture medium for 4 hours at concentrations of 0, 3.1, 6.3, 12.5, 25.0, 50.0, and 400 μg/mL in the absence or the presence of metabolic activation in the first experiment or for 24 hours at concentrations of 0, 3.1, 6.3, 12.5, 25.0, 50.0, and 400 μg/mL in the absence of metabolic activation in the second experiment. There was no evidence of an y c yt otoxicity and n o significant increase in mutant frequency over that of the solvent controls. In an in vitro test on the induction of micronuclei in cultured V79 Chinese hamster cells, V79 cell cultures were treated with Pigment Red 112 dissolved in DMSO, for 4 or 24 hrs without activation, and for 4 hr s with activation. The preparation int erval was 24 hrs in all tests. In order to assess the toxicity a cell Proliferative Index (PI) was calculated. Onl y those cultures which showed a PI > 1.3 were evaluated. The test article was found to cause no significant increases in micronucleus induction in cultured V79 cells without or with activation at the concentrations that were tested. However, the highest concentration tested with activation was well below a precipitating concentration, and no data were presented to indicate that higher co ncentrations were cy totoxic. Although the full stud y reports were not available, the IUCLID Chemical Safety Report for C.I. Pigment Red 112 illustrates two additional Bacterial Reverse Mutation Assays which support the studies presented above. In both studies no evidence of mutagenicity or cy totoxicity was seen with and without metabolic activation in S. ty phimurium TA 1535, 1537. 98, or 100 or E.coli WP uvr A. No n e ur op atholo g i ca l c h an g e s o r e f fe c ts w er e rep o rt e d in any of the studies. The agency does not believe C.I. Pigment Red 112 would be neurotoxic
. No carcinogenicit y studies are available for C.I. Pigment Red 112 . Although DEREK modeling results indicate “plausible” alerts for carcinogenicit y, this is unlikely

to be the case because C.I. Pigment Red 112 is not expected to be absorbed in the body based on the phy sical/chemical properties of the chemical . Man y azo d yes contain benzidine in their structure and this human bladder carcinogen may be released upon azo reduction of the dy e. In the case of C.I . Pigment Red 112 , azo reduction is likely to occur at low levels following oral ingestion limiting the potential fo r the formation of genotoxic and carcinogenetic metabolites. Similarly , all mutagenicity studies on C.I . Pigment Red 112 were negative which makes it unlikely that C.I . Pigment Red 112 would be carcinogenic.17

Available toxicity

studies indicate that C.I. Pigment Red 112 has a very low acute and subchronic toxicity . No endpoint of concern was identified for any of the acute studies conducted. In addition, no endpoint of concern was determined in the 28
- day study up to the limit dose of 1000 mg/kg/day (highe st dose tested). Since no endpoint of concern was identified in acute and subchronic studies and because C.I . Pigment Red 112 is not expected to be absorbed by the body based on the phy sical/chemical properties, a quantitative risk assessment for C.I. Pigm ent Red 112 was not performed. As part of its qualitative assessment, the Agency did not use safety

factors for assessing risk, and no additional safety factor is needed for assessing risk to infants and children. Furthermore, C.I. Pigment Red 112 will be used at a maximum of 10% w/w in seed treatments only . It is expected to be tightl y bound to soil so the likelihood of consumers being exposed to C.I . Pigment Red 112 as an inert ingredient through foliar ingestion or water consumption is nominal. Although dietary

exposure to C.I . Pigment Red 112 may occur from eating foods grown from seeds treated with pesticide formulations containing this inert ingredient and drinking water containing runoff from soils containing the treated seeds, the amount of residue that a consumer would be exposed to would be relativel y sma ll because C.I. Pigment Red 112 is expected to be tightl y bound to soil and therefore, it is unlikely

the pigment would be taken up by the plant and subsequently available for consumption through e ating the plant. Seeds that have been chemically treated may

NOT be used for food, feed, or oil processing. In addition, runoff is unlikely based on the low water solubility and soil binding of the pigment. There are no other known food uses for C.I. Pigme nt Red 112
. The proposed use of C.I. Pigment Red 112 as a seed treatment/dye under 40 CFR 180.920 is not expected to result in residential exposure to this chemical. In addition, there are no toxicological effects of concern in available studies and ther efore, it is not necessary to conduct a quantitative assessments of residential (non
- occupational) exposures and risks. Similarly , a quantitative occupational risk assessment is not necessary
. Based on ecological (i.e., earthworm , water flea , and algae) and fate data the a gency concluded that C.I
. Pigment Red 112 does not pose an ecological risk to terrestrial or aquatic species. C.I. Pigment Red 112 is a solid material with a low solubility in water and is not expected to volatilize. Based on the results of the Fugacity

Model C.I. Pigment Red 112 ’s highest affinity would be for sediment (52%) and soil (47%). As mentioned above, as a result of the low water solubility coupled with the projected partitioning of C.I. Pigment Red 112 , it is expected to be bound to soil and therefore, not expected to leach into ground water or be found in runoff. Although C.I. Pigment Red 112 may persist in soils, due to the small amount of pigment used compared to the weight of the seeds used per acre, deposition in soil is not expected to significantly impact soil organisms. Bioaccumulation is also expected to be low. Based on the evaluated toxicity studies coupled with the expected exposure from the use of these chemicals as inert ingredients in pesticide products, EPA concludes that there is a reasonable certainty that no harm will result to the general population, including infants and children, from aggregate exposur e to residues of C.I . Pigment Red 112 when used as an inert ingredient in pesticide formulations for seed tre atment at a maximum of 10% w/w under 40 CFR 180.92 0
.18 B ibliograph y Az , R. (2005) Determination of the Solubility of P.R. 112 in Water and Octanol. Unpublished study prepared b y Clariant
. Study I D Number: 0 019Ce
. November 1, 200 5 **. (MRID 49336101 )**  ChemIDP lus. (2013) U.S. National L ibrary of Medicine. National Institutes of Health, Department of Health & Human Services. Retrieved on January 2, 2014 from http://chem.sis.nlm.nih.gov/chemidplus Clariant Corporation . (2010) I U C L ID C h e m i ca l S a f

e t y R eport for 3
- h y

d ro x y - N
- (o
- tolyl)
- 4
- [ (2,4, 5
- tr ichlo r oph e n y

l ) azo ] n a phthal e n e - 2
- carbo x am i de; CAS RN 653 5
- 46
- 2. J u l y 1 4, 2010
. **(MRID 49242001**  **)** Dy nterova, A. (2007) (Inert Ingredient) Acute Dermal Toxicity (in Rats). Unpublished study prepared by Research Institute for Organic Sy ntheses. Project Number: 07122. June 15, 2007 **. (MRID 49148202)** Ehling, G. (2002) (Inert Ingredient) Guinea Pig Sensitization (Buehler Test). Unpublished study prepared b y Aventis Pharma Deutschland GmbH. Project Number: PT02
- 0042. June 27, 2002. **(MRID 4**  **9148206)** Environment Canada, Health Canada (2013) Draft Screening Assessment: Aromatic Azo and Benzidine
based Substance Grouping, Certain Monoazo Pigments. November 2013. Retrieved on February 12, 2014 from http://www.ec.gc.ca/ese
- ees/default.asp?lang=En &n=9C4DA306
- 1 Hall, C. (2009) In vitro Micronucleus Test in Chinese Hamster V79 Cells with Pigment Red 112. Unpublished study

prepared by Harlan Cy

totes t Cell Research Gmbh . Project Number: 1282200. September 30, 2009. **(MRID 49148208)** Krome, K. (2010) Pi gment Red 112 Earthworm (Eisenia fetida), Effects on Reproduction. Unpublished study prepared by

Dr. U. Noack
- Laboratorien. Study Number RBN13439. March 5, 2010. **(MRID 49148213)**  Levine, W. (1991) Metabolism of Azo Dy es: Implication for Detoxification and Activation. Drug Metabolism Reviews 23(3&4): 235
- 309. **(MRID 49242002)** Meurer, K. (2008) Local Ly mph Node Assay

(LLNA) in Mice with Pigment Red 112. Unpublished study prepared by

RCC Cy totest Cell Research Gmbh. Project Number: 1131902. January 25, 2008. **( MRID 49148205)**  Muller, W. (1996) (Inert Ingredient): Bacterial Reverse Mutation Test (Standard Plate Test) and Prival Modification (Preincubation Test). Unpublished study prepared by

Hoechst Aktiengesellschaft. Project Number: 96.0210. April 10, 1996. **(MRID 49148207)** Noack, M. (2010) Pigment Red 112 Daphnia magna Reproduction Test, Limit Test, Semi
- Static, 21 d. Unpublished study

prepared by

Dr. U. Noack
- Laboratorien. Study Number DRE13439. January 3, 2010. **(MRID 49148212)**19

Rupprich, N.; Weigand, W. (1 983) Permanent Red FRG Test of the Acute Oral Toxicity in the Male and Female Wistar
- Rats. Unpublished study prepared by Hoechst Aktiengesellschaft. Project Number: 83.0401 . July

27, 1983
. **(MRID 49148201)** Rupprich, N.; Weigand, W. (1983) (Inert In gredient) Test for Acute Dermal Irritation/Corrosion on the Rabbit. Unpublished study prepared by Hoechst Aktiengesellschaft. Project Number: 83.0375. July 21, 1983
. **(MRID 49148203**  **)** Rupprich, N.; Weigand, W. (1983) (Inert Ingredient) Test on Acute Ey e Irritation/Corrosion ( in Rabbits). Unpublished study prepared by Hoechst Aktiengesellschaft. Project Number: 83.0376. July 21, 1983
. **(MRID 49148204**  **)** Scheerbaum, D. (2010) Pigment Red 112 Alga, Growth Inhibition Test with Desmodesmus subspicatus, 72 h. Unpublished study

prepared by

Dr. U. Noack
- Laboratorien. Project Number: 090707CP. March 11, 2010. **(MRID 49148211)** Ullmann, L . (2002) (Inert Ingredient): Local Ly mph Node Assay

(LLNA) in Mice (Identification of Contact Allergens). Unpu blished study

prepared by

RCC Umweltchemie Ag. Study Number: 856583. January 8, 2002
. **(MRID 49148214**  **)** van Otterdijk, F. (2008) Repeated Dose 28
- Day Oral Toxicity

Study with Pigment Red 112 by Daily Gavage in the Rat. Unpublished study prepared by

Notox B. V. Project Number: 487395. June 2, 2008. **(MRID 49148210)** Wollny , H. (2008) Gene Mutation Assay in Chinese Hamster V79 Cells in vitro (V79/HPRT) with Pigment Red 112. Unpublished study prepared b y RCC Cy

totest Cell Research Gmbh. Project Number: 1154200. June 5, 2008. **(MRID 49148209)**