Reaction mass of 2-Piperazin-1-ylethanol and Piperazine-1,4-diethanol and Piperazine

Endpoint summary

Description of key information

No studies are available for hydroxyethylpiperazine mixture. Studies concerning acute toxicity are available for the components of the mixture. These were used as read across in order to assess the acute toxicity of hydroxyethylpiperazine mixture.
Oral: LD50 = 4244 mg/kg bw (CAS 103-76-4, similar to OECD 401, BASF 1968)
LD 50 = 2610 mg/kg bw (CAS 110-85-0, similar to OECD 401, BASF 1980)
LD50 > 3200 mg/kg bw (CAS 122-96-3, similar to OECD 401, BASF 1968)
Inhalation: LC0 = 1.61 mg/L (CAS 110-85-0, IHT Test, no mortality, BASF 1968)
Dermal: LD50 = 8300 mg/kg bw (CAS 110-85-0, similar to OECD 402, Huntsman 1981)

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Acceptable report which meets basic scientific principles. Hydroxyethylpiperazine is the major constituent (ca. 40%) of the test substance.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

yes

Remarks:

(weight variations in animals exceed 20% of the mean weight, observation period of 7 days, no details on animal husbandry)

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

not specified

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: young adult laboratory rats
- Weight at study initiation: 100 - 224 g

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

(doubly distilled)

Details on oral exposure:

MAXIMUM DOSE VOLUME APPLIED: 10-20 mL/kg bw
The doses were administered as aqueous solutions of 8% (800 cm³/kg), 20% (1600 cm³/kg) and 30% (3200, 4000 and 5000 cm³/kg) test substance.

Doses:

800, 1600, 3200, 4000 and 5000 cm³/kg (849, 1698, 3396, 4244, 5305 mg/kg bw - conversion in mg/kg is based on the density: d= 1.06 g/cm³)

No. of animals per sex per dose:

5

Control animals:

not specified

Details on study design:

- Duration of observation period following administration: 7 days
- Frequency of observations: daily
- Frequency of weighing: before the start of the study
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs

Sex:

male/female

Dose descriptor:

LD50

Effect level:

ca. 4 244 mg/kg bw

Based on:

test mat.

Remarks on result:

other: Corresponds to 4000 cm³/kg; the mg/kg was calculated on the density d: d= 1.061 g/cm³. 1 male/3 females died within the first 24 h post exposure in this dose group.

Mortality:

5305 mg/kg bw: All females died within the first 24 h and 3 males died within 48 h.
4244 mg/kg bw: 1 male and 3 females died within the first 24 h.
3396 mg/kg bw: 1 male and 2 females died within the first 24 h.
1698 mg/kg bw: 1 animal died within 7 days.

Clinical signs:

5305 and 4244 mg/kg bw: high stepping gait, motor excitation, intermittent respiration, abdominal position and paresis (hindlimb), immediately after application; piloerrection, nose and eyes with reddish crusts, intermittent respiration, apathy and squatting posture until including day 3. No clinical signs and findings in the surviving animals from the fifth post observation day onward.

3396 and 1698 mg/kg bw: irregular respiration and squatting posture from hour 2 until hour 4 after application. Irregular respiration and piloerection until day 4. No clinical signs and findings in the surviving animals from the fifth post observation day onward.

Body weight:

no data

Gross pathology:

Animals that died:
5305 mg/kg bw: smeared fur (snout and anogenital region), dilatation of the stomach and intestine (partly filled with blood and liquid content).
4244 mg/kg bw: smeared fur (snout); dilatation of the stomach and intestine (partly filled with blood and liquid content).
3396 mg/kg bw: smeared fur (snout and anogenital region), diarrhea.
Animals examined at termination of the study: Organs without particular findings.

Mortality:

Dose (mg/kg)	Conc.(%)	dead within 1h	dead within 1 day	dead within 2 days	dead within 7 days
5305	30	0/10	6/10	8/10	8/10
4244	30	1/10	4/10	4/10	4/10
3396	30	0/10	3/10	3/10	3/10
1698	20	0/10	0/10	0/10	1/10
849	8	0/10	0/10	0/10	0/10

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

4 244 mg/kg bw

Quality of whole database:

Studies for all constituents of the test substance are available which show all similar results.

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Acceptable study report which meets basic scientific principles. Hydroxyethylpiperazine is the major constituent (ca. 40%) of the test substance.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Version / remarks:

(adopted on 1981, 12th may; inhalation hazard test)

Deviations:

yes

Remarks:

(animals were observed for only 7 days, only 3 animals for each exposure time were used, no details only animal husbandry, exposure period of 8 hours, concentration of test substance in air mixture was not determined)

GLP compliance:

no

Test type:

other: inhalation hazard test (IHT)

Species:

rat

Strain:

not specified

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 172 g (mean)

Route of administration:

inhalation: vapour

Type of inhalation exposure:

not specified

Vehicle:

air

Details on inhalation exposure:

The test demonstrates the toxicity of an atmosphere saturated with vapours of the volatile components of the test substance at the temperature chosen for vapour generation (20°C). 3 rats per sex were exposed to the vapours, generated by bubbling 200 L/h air through a substance column of about 5 cm above a fritted glassdisc in a glass cylinder for 8h. The documentation of clinical signs was performed over a period of 7 days. In order to verify the results, the test was repeated once with new groups of animals.

Analytical verification of test atmosphere concentrations:

no

Duration of exposure:

8 h

Concentrations:

In the study report and the raw data no substance loss but an increase in substance weight was recorded.

No. of animals per sex per dose:

3

Control animals:

not specified

Details on study design:

- Duration of observation period following administration: 7 days
- Frequency of observations: daily
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight

Sex:

male/female

Dose descriptor:

other: Inhalation Risk Test

Based on:

test mat.

Exp. duration:

8 h

Remarks on result:

other: No mortality occurred.

Mortality:

No mortality occurred.

Clinical signs:

other: No clinical signs observed.

Body weight:

The treated animals gained weight (mean weight at study end: 199 g).

Gross pathology:

In one animal, chronic bronchitis and bronchiectasis in the right superior lobe of the lung were observed. Other animals were negative.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Quality of whole database:

Studies for all constituents of the test substance are available which show all similar results.

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

The study conduct was guideline compliant, however with some deviations. Only 4 animals were used per dose (2 males, 2 females) instead of 5/sex; the skin was abraded, which might have an impact on the permeability of the skin; no purity data were given. Piperazine is one constituent (ca. 15%) of the test substance.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

yes

Remarks:

purity not reported, skin is abraded, number of test animals is different

Principles of method if other than guideline:

OECD guideline says: at least 5 animals of the same sex per dose group. In this study: 4 animals (2 males, 2 females) are used per dose group.
OECD guideline says: care should be taken to avoid abrading the skin, which could alter its permeability. In this study: the skin of the rabbits were abraded by making 4 epidermal incisions with a clean needle through the stratum corneum.

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

no

Species:

rabbit

Strain:

New Zealand White

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Perfection Breeders, Inc., Douglassville, Pennsylvania
- Age at study initiation: adult
- Weight at study initiation: 2.010-2.250 kg
- Housing: individually in cages sized in accordance with the "Guide for the Care of Laboratory Animals" of the Institute of Laboratory Resources, National Research Council.
- Diet: Wayne Rabbit Ration, ad libitum
- Water: fresh tap water, fit for human consumption, ad libitum
- Acclimation period: 5 days

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

Type of coverage:

occlusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure: trunk (clipped free of fur)
- % coverage: min. 10%
- Type of wrap if used: rubber dam and an ace bandage


REMOVAL OF TEST SUBSTANCE
- Washing (if done): washing was done
- Time after start of exposure: 24 h


TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 8 g/kg bw in the preliminary test; 7, 9 and 10 g/kg bw in the main test.

Duration of exposure:

24 h

Doses:

Preliminary test: 8000 mg/kg bw
Main test for LD50 determination: 7000, 9000 and 10000 mg/kg bw

No. of animals per sex per dose:

4 animals per dose (2 male, 2 female)

Control animals:

not required

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: 30 min, 2 and 4 hours after 24 h period of exposure and twice daily thereafter for 14 days.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, dermal signs

Statistics:

According to the method of Litchfield and Wilcoxon (1949) JPET 96: 99-114.

Preliminary study:

Ten animals were treated with 8000 mg/kg bw; 3/10 animals died.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

8 300 mg/kg bw

95% CL:

7 300 - 9 400

Mortality:

Main test:
7000 mg/kg bw: 1/4 animal died.
9000 mg/kg bw 2/4 animals died.
10000 mg/kg bw: 4/4 animals died.

Clinical signs:

Cyanosis, diarrhea, salivation, mild convulsions, bleeding from mouth and nose, ataxia, loss of righting, abnormal stance, catatonia, ptosis, and decreased activity were reported. Necropsy of dying animals on study revealed black skin discoloration at application site, with hemorrhage below skin layers and skin necrosis.

Body weight:

Normal weight gain was observed in all animals.

Gross pathology:

Terminal necropsy revealed a discoloration of the tissue at application site, mottled kidneys and patchy lungs ( no visible lesions except signs of skin destruction and hemorrhage at application site).

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Based upon the observations made in the Acute Dermal Toxicity Test in Rabbits, LD50 was 8300 mg/kg bw (95% confidence limits: 7300 - 9400 mg/kg bw).

Executive summary:

In an acute dermal toxicity study performed according to OECD 402, but with deviations, an LD50 value of 8300 mg/kg bw was found.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

8 300 mg/kg bw

Quality of whole database:

The study was done pre-GLP but it is well documented and similar to OECD 402. No data are available for the remaining constituents of the test substance.

Additional information

No studies are available for hydroxyethylpiperazine mixture. Studies concerning acute toxicity are available for the components of the mixture (piperazine CAS 110-85-0, 2-piperazin-1-ylethanol CAS 103-76-4 and piperazine-1,4-diethanol CAS 122-96-3). These were used as read across in order to assess the acute toxicity of hydroxyethylpiperazine mixture.

 

Oral: In an acute oral toxicity study with piperazine (CAS 110-85-0, similar to OECD 401, BASF, 1980), groups of fasted Sprague-Dawley rats (5/sex) were given a single oral dose of the test substance in 0.5% CMC in water at 3830, 2610, 1780, 1210 and 1000 mg/kg bw and observed for 14 days. In the highest dose group all animals died. At 2610 mg/kg bw 2/5 males and 3/5 females and in the lowest dose group 1/5 males died whereas in the other dose groups no mortality occurred. Dyspnoea, apathy, abnormal position, staggering, tremor, scrubby coat, lacrimation, yellow colored urine and a bad general condition where the clinical signs seen in animals treated with the test substance. At autopsy, deceased animals showed an acute right heart dilatation and acute congested vessels, the stomach was filled atonic with liquid content, the glandular stomach was diffuse reddened, the intestine also showed an atonic with bloody diarrheic content and the lung showed an acute distension of low grade. These results lead to an LD50 of ca. 2610 mg/kg bw.

In a further study with 2-piperazin-1-ylethanol (CAS 103-76-4, similar to OECD 401, BASF, 1968) similar results were found. 5 animals per sex and dose (849, 1698, 3396, 4244, 5305 mg/kg bw) were tested by single oral application of the test substance. In this study a LD50 of ca. 4244 mg/kg bw was concluded.

Single oral application of piperazine-1,4-diethanol (CAS 122-96-3, similar to OECD 401, BASF, 1968) showed no dead animals up to 3200 mg/kg bw. Therefore, the LD50 for this substance is greater than 3200 mg/kg bw.

 

In conclusion, all three constituents of the hydroxyethylpiperazine mixture show low or no acute oral toxicity. As the hydroxyethylpiperazine (CAS 103 -76 -4) is the major constituent (ca. 50%) of the test substance a LD50 of 4244 mg/kg bw is assumed for hydroxyethylpiperazine mixture.

 

Inhalation:                                                                         

There are two inhalation hazard tests with piperazine (CAS 110-85-0, IHT test similar as described in OECD 403, BASF, 1964 and 1980) available. Groups of 3 rats per sex were exposed sequentially to the vapours, generated by bubbling 200 L/h air through a substance column of about 5 cm above a fritted glass disc in a glass cylinder for 8 or 7 hours. No analytical determination of the atmosphere concentrations was performed. The nominal concentration was calculated 1.61 mg/L or 0.57 mg/L, respectively. No animals died during the exposure time and the 7 day observation time post exposure. In the first test with the higher concentration slight mucosal irritation was reported. Body weight gain was normal and no abnormalities were detected in the gross pathology.

Since no animal died during an eight or seven hour exposure, it is assumed that there is no likely inhalation hazard under normal condition of exposure.

In a further study with 2-piperazin-1-ylethanol (CAS 103-76-4, IHT, BASF, 1967) similar results were found. 6 animals per sex were exposed for 8 hours in an inhalation hazard test with the test substance. A concentration could not be calculated but no animals died during exposure and the 7 days exposure period. No clinical signs and normal body weight gain were observed.

In an inhalation hazard test with piperazine-1,4-diethanol (CAS 122-96-3, IHT, BASF, 1968) for 8 hours no dead animals were found until 7 days post exposure. No clinical signs, normal body weight change and no abnormalities in organs were detected.

 

In conclusion, since each constituent of hydroxyethylpiperazine mixture showed no mortality in an inhalation hazard test, it can be stated that the hydroxyethylpiperazine mixture has an unlikely inhalation hazard under normal condition of exposure.

 

Dermal:

In an acute dermal toxicity study (similar to OECD 402, Huntsmann, 1981), groups of adult rabbits (2/sex) were dermally exposed to undiluted piperazine (CAS 110-85-0) for 24 hours to at least 10% of body surface area at 7000, 9000 and 10000 mg/kg bw. Animals then were observed for 14 days. Before application of the substance the skin of the rabbits were abraded. At 10000 mg/kg bw all animals died, at 9000 mg/kg bw 2/4, and at 7000 mg/kg bw 1/4 animals died. Normal body weight gain was found in this study. The animals showed following clinical symptoms: cyanosis, diarrhea, salivation, mild convulsions, bleeding from mouth and nose, ataxia, loss of righting, abnormal stance, catatonia, ptosis, and decreased activity. Necropsy of dying animals on study revealed black skin discoloration at application site, with hemorrhage below skin layers and skin necrosis. From this result a LD50 of 8300 mg/kg bw is calculated.

As only a study on piperazine is available the result from this constituent is taken as a worst case assumption for the purpose of hydroxyethylpiperazine classification, regarding the dermal route.

Justification for selection of acute toxicity – oral endpoint
Study from the major constituent of the test substance.

Justification for selection of acute toxicity – inhalation endpoint
Study for the major constituent of the test substance.

Justification for selection of acute toxicity – dermal endpoint
Only one study available.

Justification for classification or non-classification

Dangerous Substance Directive (67/548/EEC)

The available studies are considered reliable and suitable for classification purposes under 67/548/EEC. As a result the substance is not considered to be classified for acute oral, dermal or inhalation toxicity under Directive 67/548/EEC, as amended for the 28th time in Directive 2001/59/EC.

 

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

The available experimental test data are reliable and suitable for classification purposes under CLP Regulation 1272/2008. As a result the substance is not considered to be classified for acute oral, dermal or inhalation toxicity under Regulation (EC) No. 1272/2008.