Registration Dossier

Administrative data

Description of key information

Acute Toxicity:


- oral: LD50 = 410 mg/kg bw (OECD 401);
- inhalation: LC50 > 4.31 mg/L air (OECD 403);
- dermal: LD50 = 2138.7 mg/kg bw (OECD 402, rabbit); LD50: > 400 and < 2000 mg/kg bw (rat); in both WoE studies performed according to OECD 402 an LD50 > 1000 mg/kg bw was estimated

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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
GLP compliance:
yes (incl. certificate)
Test type:
fixed dose procedure
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River laboratories, Inc., Wilmington Massachusetts
- Age at study initiation: 6-10 weeks
- Weight at study initiation: 180-334 g
- Fasting period before study: yes
- Housing: individually
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): no data
- Acclimation period: min 5 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3°C
- Humidity (%): 30-70%
- Photoperiod (hrs dark / hrs light): 12h light, 12h dark


IN-LIFE DATES: From: july 14, 1992 To: july 17, 1992 (dose-finding study);
IN-LIFE DATES: From: sept 10, 1992 To: sept 24, 1992 (difinitive LD50 study)
Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
no vehicle
Doses:
dose range finding study: 500, 2500, 5000 mg/kg
definitive LD50 study: 320, 630, 1000 mg/kg
No. of animals per sex per dose:
5 rats per sex per dose
Control animals:
other: not required according to OECD guideline 401
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: 1h, 4h and 24h after dosing + daily until day 14
- Necropsy of survivors performed: yes
- Other examinations performed: incidence of clinical signs, toxic signs, mortality; body weight, gross necropsy findings
Statistics:
using the method of Litchfield and Wilcoxon via the Innovative Programming Associates, LABCAT Module Version 4.24
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
377.1 mg/kg bw
95% CL:
203.3 - 699.3
Key result
Sex:
male
Dose descriptor:
LD50
Effect level:
442.7 mg/kg bw
95% CL:
322.8 - 607.1
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
410 mg/kg bw
95% CL:
288.1 - 584
Mortality:
320 mg/kg: male 1/5 dead (day6); female 2/5 dead (day1 and day3)
630 mg/kg: male 4/5 dead (2x day1, day3; day6); female 4/5 dead (2x 3h after dosing, 2x day6)
1000 mg/kg: male 5/5 dead (3x 3h after dosing, 2x on day 1); female 5/5 dead (3 x 3h, 2x day 1)
5/5 died between 6h and 24h after exposure (dosage: 2.5 ml/kg)
1/5 died between 6h and 24h after exposure (dosage: 1.25 ml/kg)
2/5 died on the 4th day after exposure (dosage: 1.25 ml/kg)
Clinical signs:
following clinical signs are observed: decreased activity, abnormal gait, abnormal stance, dyspnea, chromodacryorrhea, diarrhea, decreased
body tone, piloerection, prostration, tremors, flaccid body tone, salivation, discolored urine, body drop and poor grooming.
Body weight:
with the exception of a slight decrease in body weight of one male in the 320·mg/kg dose level, there was an apparent increase in body weight of all surviving animals.
Gross pathology:
Necropsy of the animals dying on study revealed fluid-filled, discolored and/or distended intestines, spleen, bladder and stomach. No visible lesions
were observed in any animal at terminal necropsy.
Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
410 mg/kg bw
Quality of whole database:
GLP and OECD Guideline study

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
27. Mar 1990 - 10. Apr 1990
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
Version / remarks:
(adopted 1981)
Deviations:
no
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
Version / remarks:
adopted 2009
Deviations:
yes
Remarks:
(limit concentration for vapours of 20 mg/L not tested)
GLP compliance:
no
Test type:
fixed concentration procedure
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Dr. K. Thomae GmbH, Biberach, Germany
- Age at study initiation: 8-9 weeks
- Weight at study initiation: male: 255 ± 7 g, female: 183 ± 7 g
- Housing: groups of five per cage
- Diet: KLIBA rat/mouse laboratory diet ad libitum
- Water: ad libitum



ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 24 °C
- Humidity (%): 30 - 70 %
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:
inhalation: vapour
Type of inhalation exposure:
nose/head only
Vehicle:
other: unchanged (no vehicle)
Analytical verification of test atmosphere concentrations:
yes
Remarks:
gas chromatography
Duration of exposure:
4 h
Concentrations:
4.31 mg/L
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: before the beginning of the test, after 7 days and at the end of the observation period.
- Frequency of observations: several times during exposure and at least once on each workday in the observation period.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight,organ weights
Statistics:
The statistical evaluation of the concentration/effect relationship was carried out on the basis of the binomial test in accordance with tables of the BASF Computer Center.
Key result
Sex:
male/female
Dose descriptor:
LC50
Effect level:
> 4.31 mg/L air
Exp. duration:
4 h
Remarks on result:
other: no mortality occured
Key result
Sex:
male/female
Dose descriptor:
LC0
Effect level:
4.31 mg/L air
Exp. duration:
4 h
Remarks on result:
other: no mortality occured
Mortality:
No mortality occured.
Clinical signs:
During exposure: animals showed immediately escape attempts, eyelid closure, accelerated respiration and after 15 min restlessness. After 2 h irregular respiration and closed eyelids were noted.

After exposure and during observation period: up to 24 accelerated respiration and ruffled fur. After 48 h males showed aggressiveness and therefor were single housed. After 3 days all animals were considered as normal.
Body weight:
The body weight gain of the male rats in the test group, compared with a historical. control collective, was not affected by the substance over the total observation period.
The body weight gain of the female rats in the test group, compared with a historical control collective, was slightly retarded in the second week of the observation period.
Gross pathology:
No pathologic findings noted.

Body weight:

 Mean body weight (in g)   male        female     
 day 0  day 7  day 14  day 0  day 7  day 14
Test group  255  280  319  183  195  207
 Historical control group  248  286  318  177  196  211

The animals exhibited distinct eye irritation at the day of exposure which repressed during the recovery period (corneal stipplings, fundus not visible). There is indication that the test substance causes local irritation to exposed tissues, including respiratory tract.

Although the limit concentration of 5 mg/L was not fully met, the test is judged sufficient to characterize the acute inhalation toxicity of the substance as virtually nontoxic.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
discriminating conc.
4 310 mg/m³
Quality of whole database:
The weight-of-evidence studies were conducted according to and equivalent or similar to an OECD Guideline.

Acute toxicity: via dermal route

Link to relevant study records
Reference
Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
GLP compliance:
yes
Test type:
standard acute method
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Iffa Credo, 69210 L'Arbresle, France
- Age at study initiation: 8 weeks
- Weight at study initiation: 281+/-4g (males) and 226+/-8g (females)
- Fasting period before study: no
- Housing: per grouop of 4 to 7 during acclimatization and individually during the study period
- Diet: ad libitum with certified pelleted diet "Rats - Mice sustenance ref. AO4 C " (U.A.R.; 91360 Villemoisson-sur-Orge, France)
- Water: ad libitum with tap water filtered by a 0.22µ filter membrane (Société Millipore, 78140 Vélizy, France)
- Acclimation period: at least 5 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22+/-3
- Humidity (%): 50+/-20
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12:12


IN-LIFE DATES: From: 15 Oct 1992 (D1) To: 29 Oct 1992 (necropsy)
Type of coverage:
semiocclusive
Vehicle:
water
Details on dermal exposure:
TEST SITE
- Area of exposure: 5x6 cm (females) and 5x7 (males)
- % coverage: approx. 10 %
- Type of wrap if used: hydrophilic gauze patch (Semes France, 54183 Heillecourt, France) maintained by an adhesive hypoallergenic aerated semi-occlusive dressing (Laboratoires de Pansements et d'Hygiène, 21300 Chenôve, France) attached to a restraining bandage (Laboratoires 3M Santé, 92245 Malakoff, France)

REMOVAL OF THE SUBSTANCE: no

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): a volume of 5 ml/kg
- Constant volume used: yes
Duration of exposure:
24 hours
Doses:
400 mg/kg (main study), 1000 and 2000 mg/kg (preliminary study)
No. of animals per sex per dose:
5 males and 5 females (main study), 1 males and 1 female (preliminary study)
Control animals:
not required
Details on study design:
- Duration of observation period following administration: 14 days (5 days for preliminary study)
- Frequency of observations and weighing: Animals were observed 15, 30 hours, 1, 2, 4 hours after treatement, then once daily during 14 days. Animals were weighted before application of DMAPA and then on days 5, 8 and 15.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, cutaneous examination, macroscopic examination of the main organ (digestive tract, heart, kidneys, liver, lings, pancreas, spleen, and any organ with obvious abnormalities.
Preliminary study:
At 2000 mg/kg, both animals (one male and one female) died on day 2. They showed no clinical signs before death.
At 1000 mg/kg, both animals showed sedation and tremors between D2 and D5 with severe cutaneous reactions (necrosis associated with oedema on D2).
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 400 - < 2 000 mg/kg bw
Mortality:
At 400 mg/kg, no mortality was observed during the observation period.
Clinical signs:
At 400 mg/kg, no clinical signs were observed. No irritation was observed.
Body weight:
The body weight gain of treated animals was normal.
Gross pathology:
The macroscopic examination of the main organs of the animais sacrificed at the end of the study revealed no apparent abnormalities.
Due to the absence of macroscopic lesions, no samples were taken for histological examinations.
Preliminary Study
Dose (mg/kg) Animals Time Clinical signs Time Cutaneous reactions
1000 Male 01 30 min None D2 Necrosis, oedema
2 hours None D3 to D5 Necrosis
4 hours None  
D2 to D5 Sedation, tremors    
Female 01 30 min None D2 Necrosis, oedema
2 hours None D3 to D5 Necrosis
4 hours None  
D2 to D5 Sedation, tremors    
2000 Female 02 30 min None    
2 hours None  
4 hours None  
D2 Death    
Female 02 30 min None    
2 hours None  
4 hours None  
D2 Death    

Main Study
Dose (mg/kg) Animals Time Clinical signs
400 all (5 males and 5 females) 15 min None
30 min None
1 hour None
2 hours None
4 hours None
D2 to D15 None

Both animals died on day 2 after administration of 2000 mg/kg (1 male and 1 female). They showed no clinical signs before death.

After administration of 1000 mg/kg, both rats showed sedation and tremors between D2 and D5 with severe cutaneous reactions (necrosis associated with oedema on D2).

At 400 mg/kg, animals showed no clinical signs nor irritation.

Under these experimental conditions, the dermal LD50 of DMAPA is between 400 mg/kg and 2000 mg/kg in rats.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
1 000 mg/kg bw
Quality of whole database:
Three study records were used for a weight-of-evidence approach, two of the studies were GLP and OECD Guideline compliant.

Additional information

There are valid in vivo data available for the assessment of the acute oral and dermal toxicity of the test item.

Oral:

In the key study in rats (validity 1) conducted according to the OECD Guideline 401 (Acute Oral Toxicity), the test substance was administered unchanged to 5 rats per sex per dose by gavage at 500, 2500 and 5000 mg/kg bw (range finding study) and 320, 630 and 1000 mg/kg bw (main study) dose levels (Huntsman, PH 402 -TX-016 -92, 1991). Animals were observed for clinical signs (1h, 4h and 24h after dosing, and daily thereafter), for mortality and body weights during the 14 -day observation period, and were necropsied at death or at the end of the observation period for gross pathology. One male (day 6) and 2 females (day 1 and day 3) died at 320 mg/kg bw, 4 males (2x day 1, day 3; day 6) and 4 females (2x 3h after dosing, 2x day 6) at 630 mg/kg bw, and 5 males (3x 3h after dosing, 2x on day 1) and 5 females (3 x 3h, 2x day 1) at 1000 mg/kg bw. The clinical signs were decreased activity, abnormal gait, abnormal stance, dyspnea, chromodacryorrhea, diarrhea, decreased body tone, piloerection, prostration, tremors, flaccid body tone, salivation, discolored urine, body drop and poor grooming. Body weights increased in all dose groups (excepting the 320 mg/kg bw, where a slight body weight decrease was observed). Fluid-filled, discolored and/or distended intestines, spleen, bladder and stomach were observed in the animals that died; no visible lesions were observed in the surviving animals at terminal necropsy. Based on the results of this study, following LD50s were determined: LD 50 (females): 377.1 (95%CL of 203.3 -699.3) mg/kg; LD 50 (males): 442.7 (95%CL of 322.8 -607.1) mg/kg. The LD50 was 410 mg/kg bw (288.1 - 584) for both sexes after acute oral administration of test substance to rats. In the supporting study (validity 2) conducted in large parts equivalent to methods described in OECD guideline 401, doses of 16, 63, 252, 1008, 1632, 2528, 4008 mg/kg bw of an aqueous solution were applied by gavage followed by a post dose observation period of 7 days. Main clinical signs observed were drowsiness and staggering. At necropsy, decomposition of abdominal cavity and haemorrhagic stomachs were observed. The LD50 for oral acute toxicity in rats was calculated as ca. 1600 mg/kg bw.

Inhalation:

In the WoE study, acute inhalation toxicity was analyzed based on OECD guideline 403, adopted 1981 (BASF AG, 13I0655/897052, 1991, reliability score: 2, not tested up to limit concentration of 20 mg/L for vapours as required according to OECD guideline 403, adopted 2009). After 4 h of exposure no mortality occurred and the LC50 was set to be over the applied dose of 4.31 mg/L air. Main clinical signs observed were immediately escape attempts, eyelid closure, accelerated respiration, irregular respiration and restlessness. No pathologic abnormalities were observed. According to the fact that no mortalities occurred in the key study based on OECD 403 (adopted 1981) a LC0 of 4.31 mg/L/4h was estimated. Additional data were available from an inhalation risk test (IRT) which meets generally accepted scientific principles (BASF AG, VI/396, 1958; reliability score 2). The inhalation of a saturated vapor-air mixture for 2 hours caused mortality. Clinical signs were dyspnoea and eyes closed, ruffled fur, apathy, crusted eyelids, corneal opacity. At necropsy, hyperemia of the renal medulla and decomposed organs. In a further IRT no mortality occurred after 8 hour inhalation of the test item (Smyth et al. 1962; reliability score: 2). Based on the vapor pressure of 5.9 hPa (20°C) and a molecular weight MW = 102.2 g/mol the calculated vapor saturation is 24.8 mg/L. In the inhalation risk test (IRT) performed according to an internal standardized testing protocol (BASF AG, VI/396, 1958), the following mortalities occurred: No animals died after 30 min exposure. After 2 h exposure, 1 of 6 animals died. In the 4 h exposure group, 2 of 6 animals and in the 8 h exposure group 4 of 6 animals died. Based on the IRT cited the test substance was probably tested at the range of vapor saturation. 50% of the animals died between 4 h and 8 h of exposure. Considering the 4 h value with 33% mortality and the calculated vapor saturation as concentration additional safety is given as already an aerosol might have been formed and the test item is labeled as corrosive to the skin – Cat. 1B (GHS). Thus, the LD 50 (IRT) was estimated to the calculated vapor saturation of 24.8 mg/L/4h and thus no classification has to be performed.

Dermal:

In the first WoE acute dermal toxicity study in rats, the test item was evaluated according to OECD guideline 402 and in compliance with GLP (Arkema/CIT 9598, 1993; reliability score: 1). After a preliminary test at doses of 1000 mg/kg bw and 2000 mg/kg bw, the test item was applied to the skin of groups of 10 male and 10 female Sprague Dawley rats at doses of 400 mg/kg bw in a semi- occlusive dressing for 24 hours. Following treatment, rats were observed daily and weighted weekly. A gross necropsy examination was performed at the time of scheduled euthanasia (day 14). At doses of 2000 mg/kg bw both animals (1 male and 1 female) died on day 2 after administration. Both showed no clinical signs before death. After administration of 1000 mg/kg bw, both rats showed sedation and tremors between day 2 and day 5 with severe cutaneous reactions (necrosis associated with oedema on day 2). At 400 mg/kg bw, animals showed no clinical signs nor irritation. The LD50 for both sexes was > 400 to < 2000 mg/kg bw. In the second WoE study conducted according to the OECD guideline 402, 5 rabbits per sex per dose level were administered 1000, 2000 or 3000 mg/kg bw unchanged test substance on the dorsal intact skin area of the animal using occlusive coverage (Huntsman, PH 422-TX-015-92, 1993, reliability score: 2, no data on test item purity). After an exposure duration of 24 hours the residual test substance was rinsed and the skin dried. Animals were observed for clinical signs, mortality and body weights during the 14-day observation period, and were necropsied at death or at the end of the observation period for gross pathology. No animals died in the low dose, 3 in the mid dose (1 male and 2 females) and all but one male in the high dose. The clinical signs were decreased activity, abnormal gait, abnormal stance, decreased muscle tone, dyspnea, diarrhea, ptosis, tremors, paralysis of hindquarters, atrophy of the hind limb, poor grooming, red urine and prostration. Apparent decrease in mean body weight was observed of both treated male and female animals. At necropsy, distended and/or fluid-filled stomach and intestines, discoloured kidneys, lungs and liver, necrosis of the skin at application site were observed for the animals that died during the observation period. Discoloured kidneys were observed in 1 male (2000 mg/kg) as well as necrosis of the skin at the application site in all animals for the animals killed at the end of the study. The LD50 was set at 2138.7 mg/kg bw for both sexes. In a previous non-guideline study in male rabbits with only limited data provided, the LD50 was approximately equal to 814.83 mg/kg bw (Huntsman 64 -85, 1964). Severe erythema and oedema and destruction of the skin followed by hard eschars formation was observed. Only a short summary report without information on purity of the test item and the used method. Therefore, the the documentation is insufficient for assessment (reliability score: 4). In both studies of Huntsman (1964 and 1991) occlusive consitions were used. Due to the corrosive property and hence damaged skin, enhanced penetration of the test substance and over prediction of the test system has to be considered. The WoE study of Arkema/CIT study (1993) performed according to OECD 402 stated a dermal LD50 > 400 - <200 mg/kg bw. Nevertheless, no mortality was observed at 1000 mg/kg bw in the preliminary test of the main study (in the 2000 mg/kg bw dose group 2 of 2 animals died and in the 400 mg/kg bw dose group all animales survived). In the second WoE study according to OECD 402 (Huntsman, 1993) a dermal LD50 of 2138.7 mg/kg bw was calculated with a 95% confidence interval from 1638 to 2805 mg/kg bw. In the 1000 mg/kg bw dose group, all of the 10 animals survived, in the 2000 mg/kg bw dose group 1 of 5 males died on day 5, and 2 of 5 females died on day 4 and day 7. In the 3000 mg/kg bw dose group, only 1 of 10 animals survived. Taken together in both WoE studies performed according to OECD 402 an LD50 > 1000 mg/kg bw was estimated and thus acute dermal Cat. 4 (GHS) is considered as appropriate.


Justification for classification or non-classification

Classification, Labeling, and Packaging Regulation (EC) No 1272/2008

The available experimental test data are reliable and suitable for classification purposes under Regulation (EC) No 1272/2008. The substance is already listed in Annex VI to Regulation (EC) No 1272/2008 and classified with Cat. 4 (H302:" Harmful if swallowed").  

Additionally, the available experimental test data for oral and dermal toxicity are reliable and suitable for the purpose of classification under Regulation (EC) No 1272/2008. As a result the substance is considered to be classified for acute oral toxicity (Cat 4, H302: "Harmful if swallowed"), and acute dermal toxicity (Cat 4, H312:" Harmful in contact with skin"), under Regulation (EC) No 1272/2008 as amended for the ninth time in Regulation (EU) No 2016/1179.