2,2',2''-nitrilotriethanol citrate

Endpoint summary

Description of key information

For the substance (CAS no. 29340-81-6), the LD50 for acute oral toxicity was determined to be > 5000 mg/kg bw.

Results for dermal acute toxicity were based on the LD50 of the two dissociation product (TEA, CAS no. 102 -71 -6) of the substance, which was determined to be > 2000 mg/kg bw.

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:

13 Feb 2017 to 8 Mar 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Version / remarks:

December 2001

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

Version / remarks:

May 2008

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Version / remarks:

December 2002

Qualifier:

according to guideline

Guideline:

other: JMAFF Guidelines

Version / remarks:

Appendix to Director General Notification, No. 12-Nousan-8147. Agricultural Production Bureau, Ministry of Agriculture, Forestry and Fisheries of Japan, November 2000, including the most recent revisions

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

yes

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source of test material: Zschimmer and Schwarz GmbH + Co. KG.
- Batch No.of test material: SEALS 2016-198-001
- Purity/Composition: UVCB

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Test item storage: At room temperature
- Stable under storage conditions until: 03 January 2018 (expiry date)
- Appearance: Clear colourless liquid (determined by Charles River Den Bosch

OTHER
Purity/Composition correction factor: Yes, according to the purity: 1.28

Species:

rat

Strain:

Wistar

Remarks:

Crl: WI(Han)

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River France, L’Arbresle, France
- Females nulliparous and non-pregnant: yes
- Age at study initiation: approximately 8-9 weeks
- Weight at study initiation: 153 to 181 g
- Fasting period before study: Animals were deprived of food overnight (for a maximum of 20 hours) prior to dosing and until 3-4 hours after administration of the test item. Water was available.
- Housing: On arrival and following assignment to the study, animals were group housed (up to 5 animals of the same sex and same dosing group together) in polycarbonate cages (Makrolon MIV type; height 18 cm.) containing sterilized sawdust as bedding material (Lignocel S 8-15, JRS - J.Rettenmaier & Söhne GmbH + CO. KG, Rosenberg, Germany) equipped with water bottles. These housing conditions were maintained unless deemed inappropriate by the Study Director and/or Clinical Veterinarian. Animals were separated during designated procedures/activities. Each cage was clearly labelled. For psychological/environmental enrichment, animals were provided with paper (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom), except when interrupted by study procedures/activities.
- Diet: Pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany) was provided ad libitum throughout the study, except during designated procedures.
- Water: Municipal tap-water, ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 to 21
- Humidity (%): 43 to 49
- Air changes (per hr): Ten or greater with 100% fresh air (no air recirculation)
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

DOSE VOLUME APPLIED
The dose volume for each animal was based on the body weight measurement prior to dosing. Dose volume (mL/kg body weight) was calculated as follows: Dose level (g/kg) * (100 / purity (%)).

DOSAGE PREPARATION
The dosing formulations were stirred continuously during dose administration.

CLASS METHOD
The dose levels were based on the OECD test guidelines and were selected from the series 5 (lowest dose level), 50, 300 and 2000 (highest dose level) mg/kg body weight. The starting dose level should be the one that is likely to produce mortality in at least some of the animals and was selected based on available toxicity data of the test item.

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

6 (in two groups of 3)

Control animals:

no

Details on study design:

EXPERIMENTAL DESIGN
The toxicity of the test item was assessed by stepwise treatment of groups of 3 females. The absence or presence of mortality of animals dosed at one step determined the next step, based on the test procedure defined in the guidelines. The onset, duration and severity of the signs of toxicity were taken into account for determination of the time interval between the dose groups. The first group was treated at a dose level of 2000 mg/kg.

MORTALITY/MORIBUNDITY CHECKS
Throughout the study, animals were observed for general health/mortality and moribundity twice daily, once in the morning and once in the afternoon. Animals were not removed from cage during observation, unless necessary for identification or confirmation of possible findings.

CLINICAL OBSERVATIONS
- Postdose Observations: Postdose observations were performed at periodic intervals on the day of dosing (at least three times) and once daily thereafter. The observation period was 14 days.
- Body Weights: Animals were weighed individually on Day 1 (predose), 8 and 15.

TERMINAL PROCEDURES
All moribund animals and animals surviving to the end of the observation period were sacrificed by oxygen/carbon dioxide procedure. All animals assigned to the study were subjected to necropsy and descriptions of all internal macroscopic abnormalities were recorded.

Statistics:

No statistical analysis was performed (The method used is not intended to allow the calculation of a precise LD50 value).

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat. (total fraction)

Mortality:

No mortality occurred.

Clinical signs:

other: Piloerection was noted for four out of six animals on Day 1 only.

Gross pathology:

No abnormalities were found at macroscopic post mortem examination of the animals.

The oral LD50 value of the test substance in Wistar rats was established to exceed 2000 mg/kg body weight. According to the OECD 423 test guideline, the LD50 cut-off value was considered to exceed 5000 mg/kg body weight.

Interpretation of results:

GHS criteria not met

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

5 000 mg/kg bw

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Data waiving:

other justification

Justification for data waiving:

other:

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

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

Adequacy of study:

key study

Justification for type of information:

See read-across record in section 13.

Reason / purpose for cross-reference:

read-across source

Key result

Sex:

not specified

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: Result read-across source CAS No. 102-71-6

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

2 000 mg/kg bw

Quality of whole database:

A reliable acute dermal toxicity studies is available for triethanolamine, the dissociation product of the substance (CAS 29340-81-9).

Additional information

Acute toxicity: oral

For the test substance (CAS 29340-81-6) an acute oral toxicity study is available. The test substance was administered by oral gavage to two consecutive groups of three female Wistar rats at a dose of 2000 mg/kg body weight. Animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed after terminal sacrifice (Day 15). No mortality occurred. Piloerection was noted for four out of six animals on Day 1 only. The mean body weight gain shown by the animals over the study period was considered to be normal. No abnormalities were found at macroscopic post mortem examination of the animals. The oral LD50 value of the test substance in Wistar rats was established to exceed 2000 mg/kg body weight. According to the OECD 423 test guideline, the LD50 cut-off value was considered to exceed 5000 mg/kg body weight.

Acute toxicity: dermal

No substance-specific data on the acute dermal toxicity of the substance (CAS 29340-81-6) are available. However, according to Article 13 of legislation EC1907/2006, in case no appropriate animal studies are available for assessment information should be generated whenever possible by means other than vertebrate animal tests, i. e. applying alternative methods such as in vitro tests, QSARs, grouping and read-across. CAS 29340-81-6 is a salt of triethanolamine (TEA, CAS 102 -71 -6) and citric acid (CAS 77 -92 -9) and is expected to dissociate into the respective triethanolammonium cation and dihydrogen citrate anion (which may subsequently undergo (partial) dissociation to monohydrogen citrate and citrate anions) upon uptake by the body. Therefore it is considered to be acceptable to derive lacking information on toxicological properties of CAS 29340-81-6 by read-across from its starting materials. Citric acid is an important intermediate of the Krebs cycle (also known as citric acid cycle), and therefore occurs naturally as a metabolite in virtually all living organisms. It is also used as a natural food preservative and a food additive. The average daily intake (ADI) is not limited according to the evaluation of Joint FAO/WHO Expert Committee on Food Additives (1973), which concluded that the substance is non-hazardous to men. Therefore the toxicological behavior of CAS 29340-81-6 is expected to be governed primarily by the toxicity of TEA.

 

For TEA, one acute dermal toxicity study is available, performed similar to OECD Guideline 402. Three rabbits were exposed to 2000 mg/kg bw TEA on the intact or abraded skin and subsequently observed for 14 days. Two substances were tested: TEA derived from NH3 (91.8% TEA) or derived from DEA (88.1% TEA), both containing about 6.5% DEA. Mild erythema was observed after 24 hours following exposure to TEA (derived from NH3) on the intact and abraded skin, while moderate erythema was observed after 24 hours following exposure to TEA (derived from DEA) on the intact or abraded skin. Within the 14 -day observation period all effects disappeared. No mortality was observed and therefore the LD50 was determined to be >2000 mg/kg bw (EPA, 1989).

Justification for selection of acute toxicity – oral endpoint 

An acute toxicity study is available for the substance (CAS 29340-81-6). The LD50 was determined to be >5000 mg/kg bw.

Justification for selection of acute toxicity – dermal endpoint 

A reliable acute dermal toxicity studies is available for the dissociation product of CAS 29340-81-6, triethanolamine. The LD50 was determined to be >2000 mg/kg bw.

Justification for classification or non-classification

Based on the results of the available studies, classification for acute toxicity is not warranted in accordance with EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation No. (EC) 1272/2008.