1,4-dihydroxy-2,2,6,6-tetramethyl piperidinium-2-hydroxy-1,2,3-propanetricarboxylate

Endpoint summary

Description of key information

The test item was tested for acute oral toxicity in a GLP study according to OECD 401 on oral gavage in rats. The LD50 was determined to be 1758 mg/kg bw rats.
The test item was tested for acute inhalation toxicity in a GLP study according to OECD 403 in rats. The LC50 was determined to be greater than 5080 mg/m³ in rats.
The test item was tested for acute dermal toxicity in a GLP and guideline study according to OECD 402 on dermal administration in rats. The LD50 was determined to be greater than 2000 mg/kg bw in male and female rabbits.

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:

1999-01-26 to 1999-03-01

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP compliant Guideline study.

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Version / remarks:

February 24, 1987

Deviations:

no

GLP compliance:

yes

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Sprague Dawley, Inc. (facility not specified), USA
- Age at study initiation: approximately seven to ten weeks
- Weight at study initiation: ranging from 238 to 300 g for males and 214 to 232 g for females
- Fasting period before study: yes
- Housing: individually in suspended stainless steel cages
- Diet: PMI Certified Rodent Chow #5002 (Purina Mills, Inc.), ad libitum
- Water: municipal tap water treated by reverse osmosis, ad libitum
- Acclimation period: minimum of five days

ENVIRONMENTAL CONDITIONS
- Temperature: 21°C to 24°C
- Humidity: 35-51%
- Air changes: 10 to 15 air changes per hour
- Photoperiod: 12-hour light / 12-hour dark cycle

IN-LIFE DATES: From: 1999-01-26 To: 1999-03-01

Route of administration:

oral: gavage

Vehicle:

other: deionized water

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 500 mg / mL (fixed)

MAXIMUM DOSE VOLUME APPLIED:
- males: 6.41 mL / kg bw
- females: 4.272 mL / kg bw

Doses:

males: 1068, 2136, 2670 or 3204 mg / kg bw
females: 534, 1068, 1602 or 2136 mg / kg bw

No. of animals per sex per dose:

5 males and 5 females

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Clinical abnormalities were recorded approximately 1, 2, 4, 8 and 24 hours post-dose and daily thereafter (days 2-14).
A general health/mortality check was performed twice daily (in the morning and in the afternoon).
Individual body weights were obtained prior to fasting (day -1), prior to dosing on day 0 and for all surviving animals on days 7 and 14.
Animals found dead after day 0 were also weighed.
Individual food consumption was recorded for all surviving animals weekly.
- Necropsy of survivors performed: yes

Statistics:

The LD50 and 95% confidence intervals were calculated separately for males and the combined sexes using a computer adaption of the method of Litchfield and Wilcoxon.

Sex:

male

Dose descriptor:

LD50

Effect level:

2 494.9 mg/kg bw

Based on:

test mat.

95% CL:

1 828.9 - 3 384.8

Sex:

female

Dose descriptor:

LD50

Effect level:

> 1 068 - < 1 602 mg/kg bw

Based on:

test mat.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

1 757.8 mg/kg bw

Based on:

test mat.

95% CL:

872.6 - 3 541.1

Mortality:

All mortality occurred by study day 1:
males (mg / kg bw : fatalities / totals): 1068 : 0/5, 2136 : 1/5, 2670 : 4/5 or 3204 : 5/5
females (mg / kg bw : fatalities / totals): 534 : 0/5, 1068 : 0/5, 1602 : 5/5 or 2136 : 5/5

Clinical signs:

other: The most notable clinical abnormalities observed during the study included decreased activity, convulsions, wobbly gait, breathing abnormalities, prostration, decreased defecation, soft stools, piloerection, apparent hypothennia, skin blue in color, hunch

Gross pathology:

The most notable gross internal findings were observed in the animals that died and included abnormal content in the digestive tract, stained mucosa in the stomach and dark red lungs. Gross intemal findings observed at necropsy on study day 14 included three incidences of gray raised area(s) on the lungs in the 2136 mg/kg bw male rats.

Other findings:

Food consumption was consistent throughout the test period.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

1 757.8 mg/kg bw

Quality of whole database:

GLP and guideline compliant study.

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2003-02-25 to 2003-05-27

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

GLP compliant Guideline study. Deficiencies when compared to latest Guideline: - Clinical monitoring: now twice per day - Body weight: now mandatory for day 0 and at least on days 1, 3 and 7 (and weekly thereafter), and at the time of death or euthanasia if exceeding day 1.

Qualifier:

according to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Version / remarks:

adopted May 12, 1981

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1300 (Acute inhalation toxicity)

Version / remarks:

adopted August 1998

Deviations:

no

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Ace Animals Inc., Boyertown, PA, USA
- Age at study initiation: 11-12 weeks
- Weight at study initiation: males 321-370 g and females 204-235 g
- Housing: singly in suspended stainless steel caging with mesh floors
- Diet: Purina Rodent Chow #5012, ad libitum
- Water: tap water ad libitum
- Acclimation period: 29 days

ENVIRONMENTAL CONDITIONS
- Temperature: 16-21°C
- Humidity: 38-56%
- Photoperiod: 12 h light / 12 h dark cycle

IN-LIFE DATES: From: 2003-02-26 To:2003-03-12

Route of administration:

inhalation: dust

Type of inhalation exposure:

nose only

Vehicle:

air

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: Mini-Nose Only inhalation Chamber, ADG Developments LTD
- Exposure chamber volume: approximately 6.7 liters (internal)
- Method of holding animals in test chamber: individually in polycarbonate holding tubes
- Source and rate of air: 28.3 liters per minute (Lpm) of compressed generator air (JUN-AIR)
- Method of conditioning air: Compressed generator air was supplied to the dust generator. An additional 3.3 Lpm of compressed mixing air, supplied using dry filtered air (WELCO), was introduced into the chamber.
- System of generating particulates/aerosols: Test item compressed to 200 lbs/sq by a Carver Lab Press Model C was packed into a DF 183 Wright Dust Container. The container was fitted witth a DF 191 Stainless Steel Cutting Blade, driven by a Dayton Model 4Z538A adiustabte speed motor.
- Method of particle size determination: eight-stage Andersen cascade impactor. Aerodynamic mass median diameter was determined graphically using two-cycle logarithmic probit axes
- Treatment of exhaust air: fed into base unit
- Temperature and humidity in air chamber: 19-21°C and 38-45 %

TEST ATMOSPHERE
- Brief description of analytical method used: gravimetric samples, collected using 25 mm glass fibre filters in a filter holder connected to a vacuum pump. The mass collected was divided by the total volume of air sampled to determine the chamber concentration. Sample airflow were measured using an Omega Flowmeter Model #FMA 5610.
- Samples taken from breathing zone: yes

TEST ATMOSPHERE
- MMAD (Mass median aerodynamic diameter) / GSD (Geometric st. dev.): 3.8 µm / 2.03

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

4 h

Concentrations:

Gravimetric chamber concentration: 5.08 mg / L.

No. of animals per sex per dose:

5 males and 5 females

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Individually prior to test substance exposure (initial) and again on days 7 and 14 (termination)
- Necropsy of survivors performed: yes

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 5.08 mg/L air (analytical)

Based on:

test mat.

Exp. duration:

4 h

Mortality:

All animals survived exposure to the test atmosphere.

Clinical signs:

other: Not noted

Body weight:

All animals gained body weight over the 14-day observation period.

Gross pathology:

No gross abnormalities were noted.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating conc.

Value:

5 080 mg/m³ air

Quality of whole database:

GLP and guideline compliant study.

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:

1999-02-01 to 1999-05-28

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP compliant Guideline study.

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Version / remarks:

adopted February 24, 1987

Deviations:

no

Remarks:

(two limit tests)

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Species:

rabbit

Strain:

New Zealand White

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Myrtle's Rabbitry, Thompson Station, TN., USA
- Weight at study initiation: ranging from 2442 to 2915 g for males and 2426 to 2820 g for females
- Fasting period before study: no
- Housing: individually in suspended stainless steel cages
- Diet: PMI Certified Rodent Chow® #5322 (Purina Mills, Inc.), ad libitum
- Water: municipal tap water, ad libitum
- Acclimation period: minimum of five days

ENVIRONMENTAL CONDITIONS
- Temperature: 21°C to 24°C
- Humidity: 33-49 %
- Air changes: 10 to 15 air changes per hour
- Photoperiod: 12-hour light / 12-hour dark cycle

IN-LIFE DATES: From: 1999-02-01 To: 1999-03-17

Type of coverage:

occlusive

Vehicle:

water

Remarks:

deionized

Details on dermal exposure:

TEST SITE
- Area of exposure: dorsal trunk area, including the scapula to the wing of the ilium and halfway down the flank on each side of the animal.
- % coverage: approximately 10% of the body surface area
- Type of wrap if used: gauze dressing, a plastic wrap (occlusive binding) and an elastic wrap, secured around the trunk with adhesive tape.

REMOVAL OF TEST SUBSTANCE
- Washing: residual test item was removed using gauze moistened with deionized water followed by dry gauze.
- Time after start of exposure: approximately 24-hours

TEST MATERIAL
- Amount(s) applied:
Limit test I: dose level 2000 mg / kg bw; active dose level 1873 mg / kg bw
Limit test II: dose level 2136 mg / kg bw; active dose level 2000 mg / kg bw

- For solids, paste formed: yes

VEHICLE
- Amount(s) applied: 1 mL of deionized water / gram test item

Duration of exposure:

approximately 24-hours

Doses:

Limit test I: dose level 2000 mg / kg bw (active dose level 1873 mg / kg bw)
Limit test II: dose level 2136 mg / kg bw (active dose level 2000 mg / kg bw)

No. of animals per sex per dose:

5 males and 5 females

Control animals:

not required

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
observations: following patch removal on study day 1 and daily thereafter (days 2-14)
weighing: prior to dosing on day 0 and on days 7 and 14

- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, food consumption

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred.

Clinical signs:

other: Feces small in size, soft/mucoid stools, fecal stain and dark material around the facial area were observed during the study. Dermal irritation was noted at the site of test article application.

Gross pathology:

No significant gross internal findings were observed at necropsy on study day 14.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

2 000 mg/kg bw

Quality of whole database:

GLP and guideline compliant study.

Additional information

Oral

A study was conducted according to OECD TG 401 acute oral toxicity adopted 24 -Feb-1978. The single-dose oral toxicity of the test item was evaluated in Sprague-Dawley rats. Groups of five male and five female rats received a single oral administration of the test article. The graded dosage levels were 1068, 2136, 2670 and 3204 mg / kg bw for males and 534, 1068, 1602 or 2136 mg / kg bw for females. A gross necropsy examination was performed on all study animals at the time of death or scheduled euthanasia (day 14). All mortality occurred by study day 1. The most notable clinical abnormalities observed during the study included decreased activity, convulsions, wobbly gait, breathing abnormalities, prostration, decreased defecation, soft stools, piloerection, apparent hypothermia, skin blue in colour, hunched posture, urine/fecal stain, partially closed eyelids, salivation, dilated pupil(s), ocular discharge and dark material around the facial area. A slight body weight loss was noted for two 534 mg/kg female rats and one 1068 mg/kg female rat during the study day 7-14 body weight interval. Food consumption was consistent throughout the test period. The most notable gross internal findings were observed in the animals that died and included abnormal content in the digestive tract, stained mucosa in the stomach and dark red lungs. Gross internal findings observed at necropsy on study day 14 included three incidences of gray raised area(s) on the lungs in the 2136 mg/kg male rats. The acute oral LD50 of the test item in the male rat was determined to be 2495 mg/kg (2336 mg/kg active). In the female rat, the oral LD50 was estimated to be greater than 1068 mg/kg (1000 mg/kg active) but less than 1602 mg/kg (1500 mg/kg active). In the sexes combined, the oral LD50 was determined to be 1758 mg/kg (1646 mg/kg active).

 

Inhalation

An acute inhalation toxicity study was conducted according to OECD TG 403 adopted 1981-05-12 with rats to determine the potential for the test item to produce toxicity from a single exposure via the inhalation (nose-only exposure) route. Ten healthy Sprague-Dawley rats (5/sex) were exposed to the test atmosphere for 4 hours. The gravimetric chamber concentration was 5.08 mg/L. Based on graphic analysis of the particle size distribution as measured with an Andersen Cascade Impactor, the mass median aerodynamic diameter was estimated to be 3.8 µm. The animals were observed for mortality, signs of gross toxicity, and behavioural changes at least once daily for 14 days following exposure. Body weights were recorded prior to exposure and again on Days 7 and 14 (termination) or after death. Necropsies were performed on all animals. All animals survived exposure to the test atmosphere and gained body weight over the 14-day observation period. All animals appeared active and healthy throughout the study. No gross abnormalities were noted in the animals when necropsied at the conclusion of the 14-day observation period. Under the conditions of this study, the single exposure acute inhalation LC50 of the test substance is greater than 5.08 mg/L in male and female rats.

 

Dermal

An acute single-dose dermal toxicity study according to OECD TG 402 adopted 1987-02-24 was evaluated on New Zealand White rabbits. Two limit tests were performed in which one group of five male and five female rabbits received a single dermal administration of the test item at a dose of 2000 mg/kg (1873 mg/kg active) body weight or 2136 mg/kg (2000 mg/kg active) body weight. Following dosing, the limit test rabbits were observed daily and weighed weekly. A gross necropsy examination was performed on all limit test animals at the time of scheduled euthanasia (day 14). No mortality occurred during the limit tests. Clinical abnormalities observed during the study included feces small in size, soft/mucoid stools, fecal stain and dark material around the facial area. Dermal irritation was noted at the site of test article application. Body weight loss was noted for one male and one female in the 2000 mg/kg group and one male in the 2136 mg/kg group during the study day 0-7 body weight interval. Body weight gain was noted for all other animals during the test period. Food consumption was consistent throughout the test period. No significant gross internal findings were observed at necropsy on study day 14. Under the conditions of this test, the acute dermal LD50 of the test item was estimated to be greater than 2136 mg/kg (2000 mg/kg active) in the rabbit.

.

Justification for selection of acute toxicity – oral endpoint
Only one GLP and guideline Study available.

Justification for selection of acute toxicity – inhalation endpoint
Only one GLP and guideline study available.

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

Justification for classification or non-classification

The test item was tested for acute toxicity on oral administration, dermal administration, and inhalation exposure. The oral acute LD50 was determined to be 1757.8 mg/kg bw. The inhalation LC50 was found to be 5.08 mg/m³ air. The acute dermal LD50 was determined to be greater than 2000 mg/kg bw. Based on the results of the acute oral toxicity study a classification as acute toxic oral cat 4, H302 (Harmful if swallowed ) and Xn, R22 (Harmful if swallowed) respectively and no classification and labelling for acute dermal and inhalation toxicity is necessary according to Directive 67/548/EEC (DSD) and Regulation (EC) No 1272/2008 (CLP, GHS) criteria.