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Administrative data

Description of key information

Oral LD50 values for Diarylide Yellow Pigment 12 are > 2000 mg/kg bw, the limit for classification. No lethality was observed after acute inhalation exposure to the maximum technically feasible test concentration of 4.4 mg/L Pigment Yellow 13 (strucutral analogue) and 0.23 mg/L Pigment Yellow 17 (strucutral analogue).

No lethality was observed after single dermal application of 1710 mg/kg bw Pigment Yellow 13 (strucutral analogue).

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records
Reference
Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 4 JULY 1983 to 1 AUG 1983
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
Guideline study (OECD TG 401)
Qualifier:
according to
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Deviations:
no
GLP compliance:
yes
Test type:
standard acute method
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Strain specification: Hoe: WISKf (SPF71)
- Source: Hoechst AG, Kastengrund, SPF-breeding
- Age at study initiation: 8 - 10 weeks
- Weight at study initiation: male mean: 187,2 g; female mean: 207,3 g
- Fasting period before study: 16 h
- Housing: 5 animals per Macrolon cage
- Diet: Altromin 1324 (Altromin-GmbH, Lage/Lippe, Germany), ad libitum
- Water: tap water, ad libitum
- Acclimation period: at least 5 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 2
- Humidity (%): 55 +/- 10
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: gavage
Vehicle:
other: sesame oil
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 12,5% w/v

MAXIMUM DOSE VOLUME APPLIED: 40 ml/kg bw
Calculated dose volume was divided into two equal fractions and applied in a one hour interval.

DOSAGE PREPARATION: test substance was suspended in vehicle
Doses:
male: 5000 mg/kg bw
female: 5000, 4000, 3150 mg/kg bw
No. of animals per sex per dose:
5
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of weighing: once per week
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, macroscopic examination
Statistics:
Probit analysis: according to method of Lindner and Weber
95% confidence interval: according to method of Fieller and Sidak
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
2 228 mg/kg bw
Remarks on result:
other: The test item contained only 47% of the submission substance, i.e. LD 50 of 4740 mg test item / kg bw corresponds to 2228 mg submission substance/kg bw.
Key result
Sex:
male
Dose descriptor:
LD50
Effect level:
> 2 350 mg/kg bw
Remarks on result:
other: No males died within the observation period. The test item contained only 47% of the submission substance, i.e. LD 50 of > 5000 mg test item / kg bw corresponds to 2350 mg submission substance/kg bw.
Mortality:
males: none of the tested animals died within the observation period
females:
- 5000 mg/kg bw: 3/5
- 4000 mg/kg bw: 1/5
- 3150 mg/kg bw: 0/5
The animals died within the first three days after application.
Clinical signs:
males:
On the day of application the animals showed a reduced spontaneous activity, one male showed spread legs and three males a crouched posture. All animals had yellow-coloured faeces on day 1 and 2 and yellow coloured skin on days 1-3. From day four on until day 14 all of the animals were free of any clinical symptoms.

females:
- 3150 mg/kg bw: All of the tested animals were without any clinical symptoms for up to two hours after application. Between two and six hours all animals showed quiet behaviour. The skin of all animals was yellow-coloured starting two hours after application and lasting until day three. After one day there occured yellow-coloured faeces. From day four on until day 14 all of the animals were free of any clinical symptoms.
- 4000 mg/kg bw: All of the tested animals were without any clinical symptoms for up to two hours after application. Between two and six hours all animals showed quiet behaviour. The skin of all animals was yellow-coloured starting two hours after application and lasting until day three. After one day there occured yellow-coloured faeces. One out of the five tested animals showed still quiet behaviour and couched posture with spread legs after one day and also displayed ruffeled fur, convulsions, constricted palpebral fissures and a lowered respiratory rate. One animal died. From day four on until day 14 the remaining four animals were free of any clinical symptoms.
- 5000 mg/kg bw: All of the tested animals were without any clinical symptoms for up to one hour after application. Three animals died. From day eight on until day 14 the remaining two animals were free of any clinical symptoms.
Body weight:
no reduction of the body weight development
Gross pathology:
male:
- 5000 mg/kg bw: no macroscopic anomalies found
female:
- 3150 mg/kg bw: no macroscopic anomalies found
- 4000 mg/kg bw: 4 animals had no macroscopic anomalies
The one animal which died during the test was eroded.
- 5000 mg/kg bw: 2 animals sacrificed at the end of the postexposure period had no macroscopic anomalies
3 animals died during the observation period:
- stomach was tightly filled with air and test substance
- intestinal tract was filled with test substance
- pancreas was rosy-coloured
- in two out of this three animals the lung and the adrenal gland were dark-coloured
- one out of the three animals displayed bright foci on the liver
Interpretation of results:
GHS criteria not met
Remarks:
Regulation (EC) no 1272/2008
Conclusions:
The testing for acute oral toxicity according OECD TG 401 yielded a median lethal dose (LD50) of 4740 mg test item/kg bw (corresponding to 2228 mg P.Y. 12/kg bw) in female Wistar-rats. The testing in male rats revealed no obvious toxicity in the tested dose, thus leading to a LD50 > 5000 mg/kg bw (corresponding to > 2350 mg Permanent-Gelb DHG/kg bw).
The test was performed with Permanent-Gelb DHG-80 which contains relevant amounts of Permanent-Gelb DHG, which is identical to the submission substance. Therefore the test results are considered adequate to fulfill the endpoint requirements.
Executive summary:

Acute oral toxicity of the test item was tested in male and female Wistar rats according to OECD TG 401. Administered dose levels were 3150, 4000 and 5000 mg/kg bw in female and 5000 mg/kg bw in male rats. Three females in the highest dose level and one female in the middle dose level died within 3 days after application. An oral LD50 of 4740 mg test item/kg bw (corresponding to 2228 mg P.Y. 12/kg bw) has been calculated. No lethality was observed in male rats.

Therefore, the test item has not to be classified for acute toxicity according to Regulation (EC) No 1272/2008.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
2 228 mg/kg bw
Quality of whole database:
reliable without restriction

Acute toxicity: via inhalation route

Link to relevant study records

Referenceopen allclose all

Endpoint:
acute toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
1978
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
Deviations:
yes
Remarks:
: 10 hrs light cycle, some exposure data are missing (e.g. MMAD (not calculated), equilibration period)
GLP compliance:
no
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
other: Tif: RAIf (SPF)
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: in house breeding, SPF breeding colony
- Weight at study initiation: 170 - 190 g
- Housing: in Macrolon cages (type 4) in groups of 10, males and females separated
- Diet: NAFAG (Gossau SG, Switzerland), ad libitum
- Water: ad libitum
- Acclimation period: at least 4 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22+/-1
- Humidity (%): 55+/-5
- Photoperiod: 10 hrs light cycle day
Route of administration:
inhalation: aerosol
Type of inhalation exposure:
nose only
Vehicle:
other: unchanged (no vehicle)
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: inhalation cylinder
- Method of holding animals in test chamber: seperate PVC tubes
- System of generating aerosols: Injecting the solid material with the help of "Grafix Exactometer injector" into an air stream which was discharged into the exposure chamber through a nozzle under a pressure of 2 atm. at a rate of 20 L/min.
- Method of particle size determination: gravimetrically
- Temperature, humidity, oxygen: 24 °C, 38%, 20 Vol.%

TEST ATMOSPHERE
- Brief description of analytical method used: Concentration was determined gravimetrically by sampling the test atmosphere through a selectron filter of 50 mm diameter and with a pore size of 0.2 µm (Schleicher and Schuell, Feldbach, Switzerland) at an air flow rate of 10 L/min. The size distribution of the particles was measured with a Cascade Impactor with selectron filters of 25 mm diameter and with pore size of 0.2 µm (Schleicher and Schuell, Feldbach, Switzerland) at an air flow rate of 17.5 L/min.
- Samples taken from breathing zone: yes

TEST ATMOSPHERE (if not tabulated)
- Particle size distribution: < 1 µm to > 7 µm
- MMAD (Mass median aerodynamic diameter) / GSD (Geometric st. dev.): not calculated
Analytical verification of test atmosphere concentrations:
yes
Remarks:
: gravimetric measurement
Duration of exposure:
4 h
Concentrations:
4250 (+/-128) mg/m³
No. of animals per sex per dose:
10
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Necropsy of survivors performed: yes
Key result
Sex:
male/female
Dose descriptor:
LC50
Effect level:
> 4 250 mg/m³ air (analytical)
Exp. duration:
4 h
Remarks on result:
other: no animals died within the 14 day observation period
Mortality:
- no death occurred
Clinical signs:
- within 3 to 4 hours after starting the exposure period rats showed dyspnoea, curved or ventral position and ruffled fur
- recovery within 4 days
Gross pathology:
- no macroscopically visible changes were found
Interpretation of results:
GHS criteria not met
Conclusions:
Exposure of male and female rats to 4250 mg/m³ test item for 4 hours did not result in the death of the animals during a 14 day observation period, resulting in a LC50 value of > 4250 mg/m³.
Executive summary:

Acute inhalation toxicity of the test item has been investigated in male and female Tif: RAIf (SPF) rats (10 per sex). They were exposed to 4250 mg test substance per cubic meter for 4 h (maximal applicable dose). All animals survived the 14 day observation period. No macroscopic visible changes were observed at necropsy at the end of the observation period, resulting in a LC50 value of > 4250 mg/m³ (4.25 mg/L) for the inhalation of aerosol.

Classification for acute inhalation toxicity is not necessary according to Regulation (EC) No 1272/2008 .

Endpoint:
acute toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 4 DEC 1989 to 18 DEC 1989
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.2 (Acute Toxicity (Inhalation))
Deviations:
no
GLP compliance:
yes
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Hoechst AG, Kastengrund, SPF breeding colony
- Age at study initiation: 8 to 10 weeks
- Weight at study initiation: males mean: 227 g; females mean: 195 g
- Housing: macrolon cages (type 4) groups of 5 respectively 2 (= control animal and animal killed 1 day post application)
- Diet: rat diet Altromin 1324 ( Altromin-GmbH, Lage/Lippe, Germany); ad libitum
- Water: Tap water; ad libitum
- Acclimation period: at least 5 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22+/- 2
- Humidity (%): 50 +/- 20
- Photoperiod (hrs dark / hrs light): 12/12


Route of administration:
inhalation: dust
Type of inhalation exposure:
nose only
Vehicle:
other: unchanged (no vehicle)
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: cylindrical plastic cage
- Exposure chamber volume: 60 L
- Method of holding animals in test chamber: each animal was in a plastic tube
- Source and rate of air: laminar air stream of 1000 L/hour at 4 bar from above
- System of generating particulates/aerosols: dustgenerator "Wright Dust Feed" of L. Adams Ltd., London, UK
- Method of particle size determination: "Anderson-Kaskadenimpaktor Mark III" of Anderson Samples Inc, Atlanta, USA
- Treatment of exhaust air: exhaust device at the basement of the exposure chamber in line with a diverse system of filters

TEST ATMOSPHERE ANALYSIS
- Brief description of analytical method used: gravimetric measurement: The test atmosphere was sucked through filters ("Experimentiertrommelgasfilter", Elster AG, Mainz-Kastel, Germany; as well as a fibre glass and a membrane filter (diameter of pores 0.65 µm) Satorius Membranfilter GmbH, Göttingen, Germany) by means of vacuum. The exhaust quantity was 3 L/min, which resulted in an air flow of 1.25 m/sec. The filters were positioned in an exsiccator 24 h before use. The filters were weighed before and after each measurement by means of electrical scales.

TEST ATMOSPHERE
- Particle size distribution: < 0.6 µm to > 10.3 µm
- MMAD (Mass median aerodynamic diameter) / GSD (Geometric st. dev.): 1.0 - 1.1/ 2.3 - 2.8
Analytical verification of test atmosphere concentrations:
yes
Remarks:
gravimetric measurement
Duration of exposure:
4 h
Concentrations:
230 mg/m³ air
No. of animals per sex per dose:
6/sex/treatment group
1/sex/control group
Control animals:
yes
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations: twice a day
- Frequency of weighing: weekly
- Necropsy of survivors performed: yes

- Control urine and blood samples were collected from control animals.
- On day 1, 2 and 7 one male and one female were sacrificed to obtain blood samples. Final sacrifice of the remaining 3 animals/sex was on day 14 of the observation period.
- Urin samples were collected starting on the day of exposure till day 7 of the observation period and the last night before sacrifice.
- To collect urine the rats were placed in metabolism cages overnight without witdrawal of food and water. For technical reasons urine was collected continously on day 6 and 7.
- Blood was collected via puncture of the retro-orbital plexus.
Key result
Sex:
male/female
Dose descriptor:
LC50
Effect level:
> 230 mg/m³ air
Exp. duration:
4 h
Remarks on result:
other: no animals died within the 14 day observation period
Mortality:
- no deaths occurred
Clinical signs:
- during the exposition: irregular breathing, narrowed palpebral fissures and female animals showed additional stilted gate
- no more clinical signs were observed in males 330 minutes p.a. and in females 450 minutes p.a.
Body weight:
- Body weight development was impaired in the first week, probably due to the frequent placement in metabolism cages.
- But all animals except for one female did excess their initial body weight at the end of the test period.
Gross pathology:
- no macroscopically visible changes were found
Interpretation of results:
GHS criteria not met
Conclusions:
Exposure of male and female Wistar rats to 230 mg/m³ test item for 4 hours did not result in the death of the animals during a 14 day observation period, resulting in a LC50 value of > 230 mg/m³.
Executive summary:

Acute inhalation toxicity of the test item has been investigated in male and female Wistar rats. They were exposed to 230 mg test substance per cubic meter for 4 h (maximal applicable dose). All animals survived the 14 day observation period. No macroscopic visible changes were observed at necropsy at the end of the observation period, resulting in a LC50 value of > 230 mg/m³ for the inhalation of dust.

Classification for acute inhalation toxicity is not necessary according to Regulation (EC) No 1272/2008 .

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LC50
4 250 mg/m³
Quality of whole database:
reliable

Acute toxicity: via dermal route

Link to relevant study records
Reference
Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1972
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
yes
Remarks:
: no details given on the duration of the exposure; no details given on the occlusion made on the application site; dose tested (1710 mg/kg bw) is lower than the recommended limit dose (2000 mg/kg bw); only 7 days postobservation period
GLP compliance:
no
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: in house breeding
- Age at study initiation: 8 weeks
- Weight at study initiation: males mean: 322 g; females mean: 210 g
- Housing: single caged
- Diet: pelleted diet (Oakes of Congleton), ad libitum
- Water: ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21+/-2
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:
not specified
Vehicle:
other: 50:50 mixture: polyethylene glycol/water
Details on dermal exposure:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 3000 mg/kg bw
- For solids, paste formed: yes (1 g of test item was mixed with 2.5 ml of vehicle)
Duration of exposure:
no data
Doses:
9.9 ml of paste per kg bw (equivalent to 3000 mg test item per kg bw)
No. of animals per sex per dose:
5
Control animals:
yes
Details on study design:
- Duration of observation period following administration: 7 days
- Necropsy of survivors performed: yes
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 3 000 mg/kg bw
Remarks on result:
other: corresponds to 1710 mg/kg bw pure Pigment Yellow 13; no animals died within the observation period
Mortality:
- no deaths occurred
Clinical signs:
- no clinical symptoms were recorded
Gross pathology:
- No macroscopically visible changes occurred at necropsy.
Interpretation of results:
GHS criteria not met
Conclusions:
The test item was not toxic to male and female Sprague-Dawley rats after dermal application, resulting in a dermal LD50 > 3000 mg/kg bw (corresponding to > 1710 mg PY13/kg bw).
The test was performed with the test item which contains relevant amounts of the submission substance. Therefore, the test results are considered adequate to fulfil the endpoint requirements.
Executive summary:

Acute dermal toxicity of the test item was tested in male and female Sprague-Dawley rats. The maximal applicable dose (3 g test item /kg bw suspended in polyethylene glycol:water 50:50 - final volume 9.9 ml; corresponding to 1710 mg PY13/kg bw) were applied to the skin. No clinical signs were observed and no animals died during the 7 day observation period, resulting in a LD50 > 3000 mg/kg bw (corresponding to > 1710 mg PY13/kg bw).

Classification for acute dermal toxicity is not necessary according to Regulation (EC) No 1272/2008.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
1 710 mg/kg bw
Quality of whole database:
reliable

Additional information

Reliable data from several guideline studies on acute toxicity after oral application are available for Pigment Yellow 12 and its structural analogues, Pigment Yellow 13, Pigment Yellow 14, Pigment Yellow 17, Pigment Yellow 55, Pigment Yellow 83, Pigment Yellow 126, Pigment Yellow 127, Pigment Yellow 170, Pigment Yellow 174 and Pigment Yellow 176 (please refer to read across justification document, IUCLID section 13). These data reveal a very low acute oral toxicity of the pigments of this category: LD50 values for all investigated test items in rats are above 2000 mg/kg bw, the upper limit for classification. Even in tests performed with much higher concentrations (up to 15000 mg/kg bw) no animals died during the post observation period.

Based on these data it is concluded that no classification for acute oral toxicity is necessary for Diarylide Pigment Yellow 12.

Acute toxicity after inhalation exposure was investigated with the strucutral analogues, Pigment Yellow 13 and Pigment Yellow 17. In the guideline study on acute toxicity after inhalation exposure performed with Pigment Yellow 17 rats were exposed to the maximum technically feasible dust concentration of 0.23 mg/L (MMAD about 1.1 μm). No lethal effects, no severe clinical symptoms indicating a life-threatening or moribund state, and no gross morphological abnormalities were observed. Similar results were obtained with Pigment Yellow 13 in two studies, which were equivalent to guideline studies. All rats exposed to aerosol concentrations of 4.2 mg/L or 4.4 mg/L for 4 h survived during the 14 - days observation period. As no lethal effects occurred at the maximum technically feasible concentrations it is concluded that the members of the Diarylide Yellow Pigments category have not to be classified for acute toxicity after inhalation exposure.

Acute toxicity after dermal application has been tested with Pigment Yellow 13 (strucutral analouge) in male and female rats which received 3000 mg/kg bw of the test item (corresponding to 1710 mg/kg bw pure Pigment Yellow 13). No lethality occurred during the 7 day observation period and no macroscopically visible changes were observed at necropsy. Taking into account that the members of this category a) show very low toxicity after oral application, b) do not induce skin irritation and c) are not expected to penetrate through the skin due to their very low water solubility and based on the data on acute toxicity after dermal application of Pigment Yellow 13 it is concluded that Diarylide Yellow Pigments have not to be classified for acute dermal toxicity.

No specific toxic effects were observed after acute oral, inhalative or dermal exposure indicative of a specific target organ toxicity. For additional information refer to read across justification, IUCLID Chapter 13.

Justification for classification or non-classification

Based on the reported study data Diarylide Yellow Pigment 12 has not to be classified for acute toxicity according to Regulation (EC) No 1272/2008 and Council Directive 67/548/EEC.

Furthermore, Diarylide Yellow Pigments have not to be classified for specific target organ toxicity – single exposure according to Regulation (EC) No 1272/2008, as no specific toxic effects were observed after acute exposure.