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Administrative data

Description of key information

Mortality, physical observations, body weight, and food consumption data, as well as gross necropsy observations did not reveal any adverse effects considered to be attributable to the administration of EAD01 at any of the dose levels. 

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records
Reference
Endpoint:
short-term repeated dose toxicity: oral
Type of information:
migrated information: read-across based on grouping of substances (category approach)
Adequacy of study:
key study
Study period:
2004
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: according to EC Directive 92/69/EEC and Regulation EC/440/2008 guideline methods under GLP conditions. This data valid as documented CCRF for this category (Acetylenic geminalic diols)
Qualifier:
according to
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity in Rodents)
Deviations:
no
GLP compliance:
yes
Species:
rat
Strain:
other: HsdBrlHan:WIST
Sex:
male/female
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on analytical verification of doses or concentrations:
The test article was weighed into pre-labelled beakers and
the requested volume of vehicle was measured using
suitable apparatus. The formulation was made up to volume
in a beaker or formulation bottle up to a calibrated line. The
formulation was placed on a Silverson to ensure
homogeneity, then divided into the requested number of
aliquots. A correction of 1.099 was used.
Duration of treatment / exposure:
Oral gavage. Daily for 30 days. Dose volume 5 mL/kg.
Frequency of treatment:
Daily
Remarks:
Doses / Concentrations:
5 mL/kg
Basis:
actual ingested
No. of animals per sex per dose:
5
Control animals:
yes
Observations and examinations performed and frequency:
Clinical Observations Twice daily (am/pm) for routine health checks and daily for
signs of ill health or overt toxicity.
Physical Examinations Weekly intervals
Body Weights Before treatment on first day of dosing, at weekly intervals
and before necropsy.
Food Consumption Weekly intervals. Calculated as g/animal/week.
Haematology All main study animals at the end of Week 4 from the
lateral caudal vein after an overnight period without food.
Clinical Chemistry# All main study animals at the end of Week 4 from the
lateral caudal vein after an overnight period without food.
In addition, blood samples were obtained from females only
in Groups 1 and 2 at necropsy from the abdominal aorta.
Functional
Observational Battery
All main study animals before initiation of treatment and
once weekly thereafter.
Observational measurements were evaluated before and
upon removal from the home cage. The animal was also
placed into a circular arena for two minutes.
In Week 4, an assessment of sensory reactivity to stimuli,
grip strength and motor activity was performed.
Sacrifice and pathology:
Necropsy All main study animals after an overnight period without
food.
Organ weights Specified organs were dissected free from fat and other
contiguous tissue and weighed before fixation.
Tissue Preservation Specified tissues were preserved in the appropriate
fixative/s
Histology Gross lesions and specified tissues from all control and high
dose main study animals.
Liver, thyroid and stomach from low and intermediate dose
animals were processed to slide stage.
Kidney from low and intermediate dose male animals were
processed to slide stage
Dose descriptor:
NOAEL
Effect level:
ca. 150 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: overall effects
Critical effects observed:
not specified
Conclusions:
EnviroGem AD01 Surfactant was orally administered at dose levels of 0, 15, 150 and
1000 mg/kg/day for at least 28 days and was well tolerated. There were no significant
clinical observations, and only minor increases in bodyweight and food consumption
at 1000 mg/kg/day. Increases in total cholesterol, globulin and total protein were of
unknown toxicological significance. Microscopic changes in the liver, thyroid, male
kidney and stomach were due to effects of the test article, but only those in the liver
and thyroid were of toxicological significance. Microscopic findings in the liver,
thyroid, male kidney and stomach of animals dosed at 15 mg/kg/day were similar to
controls.

The liver weight changes and the severity of the microscopic changes in the liver,
thyroid and kidney can be associated with the increased metabolism of EnviroGem
AD01 Surfactant, and therefore, 150 mg/kg/day was considered to be The
No-Adverse-Effect Level (NOAEL).
Executive summary:

The No-Observed-Adverse-Effect-Level (NOAEL) for EnviroGem AD01 Surfactant when administered to rats by oral gavage for 28-days is 150 mg/kg/day. Toxicologically significant effects were seen in the liver and thyroid of rats dosed at 1000 mg/kg/day.


The purpose of this study was to determine the toxicity of the test article, EnviroGem AD01 Surfactant, following oral (gavage) administration to the rat for at least 28 days.

Male and female HsdBrIHan:WIST rats were assigned to four groups (five animals/sex/group). Each group received dose preparations at dose levels of 0, 15, 150 or 1000 mg/kg of body weight at a dose volume of 5 mL/kg.

A pilot study run before the start of the study confirmed that in three male and three female rats dosed at 1000 mg/kg for five days no severe toxicity was seen.

Assessment of toxicity, was based on mortality, clinical observations, clinical and anatomic pathology evaluations, clinical pathology and functional observational battery and locomotor activity.

There were no decedents during the study.

Total cholesterol was seen to increase in a dose-related manner in both males and females. Animals dosed at 1000 mg/kg/day recorded increases in total cholesterol compared to control of 71% for males (p<0.001) and 63% for females (p<0.01). Animals dosed at 150 mg/kg/day also showed increases in both sexes (males 29% and females 25%).

A dose-related increase in globulin was seen in males dosed at 150 mg/kg/day (8%) and 1000 mg/kg/day (25%, p<0.01). Group mean globulin was also increased in a dose-related manner in females. This was reflected in an increase in total protein values in these animals.

Group mean liver weights adjusted to overall mean body weight increased in males and females dosed at 150 mg/kg/day and a greater magnitude in animals dosed at 1000 mg/kg/day.

Macroscopically, there was large or mottled liver, and pale or mottled kidney in some treated animals.

Microscopically, in high dose animals, there was hepatocyte hypertrophy in the liver; increased follicular cell hypertrophy in the thyroid; increased hyaline droplets (males only) and focal nephropathy in the kidney; and minor squamous cell hyperplasia in the fore stomach, due to effects of the test article. Liver hepatocyte hypertrophy and increased hyaline droplets in the male kidney were also seen in intermediate dose animals.

Microscopic findings in the liver, thyroid, male kidney and stomach of low dose animals were similar to controls and therefore this dose was considered to be the overall no observable effect level (NOEL).

In conclusion, EnviroGem AD01 Surfactant was orally administered at dose levels of 0, 15, 150 and 1000 mg/kg/day for at least 28 days and was well tolerated. There were no significant clinical observations, and only minor increases in body weight and food consumption at 1000 mg/kg/day. Increases in total cholesterol, globulin and total protein were of unknown toxicological significance. Microscopic changes in the liver, thyroid, male kidney and stomach were due to effects of the test article, but only those in the liver and thyroid were of toxicological significance. Microscopic findings in the liver, thyroid, male kidney and stomach of animals dosed at 15 mg/kg/day were similar to controls.

The liver weight changes and the severity of the microscopic changes in the liver, thyroid and kidney can be associated with the increased metabolism of EnviroGem AD01 Surfactant, and therefore, 150 mg/kg/day was considered to be The No-Observed-Adverse-Effect Level (NOAEL).

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
150 mg/kg bw/day
Study duration:
subchronic
Species:
rat
Quality of whole database:
The key study is GLP compliant and is of high quality, Klimisch score = 1. It is done according to OECD guideline 407.

Additional information

Mortality, physical observations, body weight, and food consumption data, as well as gross necropsy observations did not reveal any adverse effects considered to be attributable to the administration of EAD01 at any of the dose
levels. NOAEL = 150 mg/kg bw. This result is supported by a 90 days test on beagles and a test 28 Day Oral (Diet) Administration Toxicity Study in the Rat on TMDD.

Surfynol compounds in the AGD category do not show significant toxicity when administered to animals over long (91 days) periods of time. The lowest NOAEL in this set of repeated dose toxicity studies is 125 mg/kg bw/day, from the OECD 422 study on S-104 in WISTAR/Han rats ranging from 42 to 48 days duration with a 14 day recovery period. A similar NOAEL is obtained from the repeated dose study of EAD01, at 150 mg/kg bw/day, from the 28-day study of EAD01 in Hsd:WISn rats.  As discussed above, it is likely that studies could be designed to generate NOAEL values which are higher than these values, as adaptive effects in the liver contributed to the selection of these current values. These studies also represent the studies with Klimisch validity scores of 1, which surpass those of older studies which predate establishment of OECD guidelines and GLP. However, the existence of 90-day studies with higher NOAEL values should be kept in mind by risk assessors. The NOAELs selected above will result in very conservative DNEL values, below those established by authoritative bodies in the United States.


Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:
This study contains data which meets the criteria set forth in Regulation EC No. 440/2008 for filling REACH endpoints. The data is generated in a reliable laboratory using established protocols. This data is valid as documented CCRF for this category (Acetylenic geminalic diols). It is supported by
three Klimisch 2 rated studies (90 days beagles and 28 respectively 90 days rats) done before GLP was established on TMDD as such.

Justification for classification or non-classification

According to the UN Globally Harmonized System of Classification and Labelling of Chemicals (GHS) Part 3 no classification required.