2-(2-butoxyethoxy)ethyl methacrylate

Endpoint summary

Description of key information

Subacute study; oral (gavage); rat (Hsd: Sprague Dawley SD), m/f (OECD guideline 422, Klimisch score: 1,GLP), with Butyldiglycol

methacrylate: NOAEL = 1000 mg/kg bw/d (HRTF, 2014).
The primary metabolite of BDGMA, the alcohol Butyldiglycol, did not show any effects of systemic toxicity towards male and female rats at doses up to 2000 mg/kg/day when administered dermally over a 13 week period.

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

Remarks:

combined repeated dose and reproduction / developmental screening

Type of information:

experimental study

Adequacy of study:

key study

Study period:

27 August to 22 October 2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

GLP, coherence between data, results and conclusions

Qualifier:

according to guideline

Guideline:

OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)

Version / remarks:

adopted on 22 March 1996

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Specific details on test material used for the study:

Identity: Butyldiglycol methacrylate
Alternative name: VISIOMER®BDGMA, BDGMA

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
A total of 90 Sprague Dawley [Crl:CD(SD)BR] rats (45 males and 45 virgin females), 6 to 7 weeks old and weighing 190 - 217 g for males and 175 - 189 g for females, were received from Charles River Italia S.p.A., Italy.
After arrival the weight range for each sex was determined and the animals were temporarily identified within the cage by means of a coloured mark on the tail. A health check was then performed by a veterinarian.
An acclimatisation period of 13 days was allowed before the start of treatment, during which time the health status of the animals was assessed by thorough observations.


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22°C +/- 2°C
- Humidity (%): 55% +/- 15%
- Air changes (per hr): 15 to 20 air changes per hour
- Photoperiod (hrs dark / hrs light): artificial light for 12 hours each day

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

The test item was administered orally by gavage at a dose volume of 5 mL/kg body weight. Control animals received the vehicle alone at the
same dose volume.

The required amount of Butyl diglycol methacrylate was suspended in the vehicle (corn oil) and brought to the final volume appropriate for each
concentration (concentrations of 20, 60 and 200 mg/mL) and kept under magnetic stirrer at room temperature prior to use and until the time of dosing of the last animal.
The formulations were prepared daily and the concentrations were calculated and expressed in terms of test item as supplied.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Prior to commencement of treatment, analysis was performed to confirm that the proposed formulation procedure was acceptable (concentration
and homogeneity). Results of the analyses were within the limits of acceptance.
The stability was found to be 24 hours at room temperature in the concentration range of 20 to 200 mg/mL.
Samples of the formulations prepared on Week 1 and last Week were also analysed to check the concentration and homogeneity. Results of the analyses were within the limits of acceptance.
Chemical analysis was carried out by the Analytical Chemistry Department at RTC according to a validated method (RTC Study No. 96370), in the range from 5 to 200 mg/mL.

Duration of treatment / exposure:

Males
Animals were dosed once a day, 7 days a week, for a minimum of 2 consecutive weeks prior to pairing and thereafter through the day before necropsy
(Day 29 - 30 of treatment).

Dose volumes were adjusted once per week for each animal according to the last recorded body weight.

Females
Animals were dosed once a day, 7 days a week, for a minimum of 2 consecutive weeks prior to pairing and thereafter during pairing, post coitum and
post partum periods until Day 3 post partum (the day before sacrifice).

Dose volumes were adjusted once per week for each animal according to the last recorded body weight.
During the gestation period, dose volumes were calculated according to individual body weight on Days 0, 7, 14 and 20 post coitum and on Day 1
post partum. Thereafter individual dose volumes remained constant.

Frequency of treatment:

once daily

Dose / conc.:

100 mg/kg bw/day (nominal)

Remarks:

Basis: nominal in corn oil

Dose / conc.:

300 mg/kg bw/day (nominal)

Remarks:

Basis: nominal in corn oil

Dose / conc.:

1 000 mg/kg bw/day (nominal)

Remarks:

Basis: nominal in corn oil

No. of animals per sex per dose:

The test item was administered orally, by gavage. The oral route was selected as it is a possible route of exposure of the test item in man.
3 groups comprised 10 males and 10 females rats received the test item at the dose levels of 100, 300 and 1000 mg/kg/day.
Each group comprised 10 male and 10 female rats.

Control animals:

yes

Details on study design:

Dose levels of 100, 300 and 1000 mg/kg/day were selected by the Sponsor based on information from a previous non GLP compliant study (2 week preliminary study) (RTC Study no.: 96020EXT).

Observations and examinations performed and frequency:

Parenteral examination
Mortality

Throughout the study, all animals were checked early in each working day in the morning and in the afternoon. At weekends and Public Holidays a
similar procedure was followed except that the final check was carried out at approximately mid-day.

Clinical signs

Once before commencement of treatment and at least once daily during the study, each animal was observed and any clinical signs were recorded.
Observations were performed at the same time interval each day, the interval was selected taking into consideration the presence of post-dose
reactions.

Clinical observations (Functional Observation Battery Tests)

Once before commencement of treatment and once a week thereafter, each animal was given a detailed clinical examination.
Each animal was removed from the home cage and observed in an open arena.
The tests included observation of changes in gait and posture, reactivity to handling, presence of clonic or tonic movements, stereotypies or bizarre
behaviour and effects on the autonomic nervous system (e.g. lachrymation, piloerection, pupil size, unusual respiratory pattern).
All observations were recorded for individual animals.

All observed parameters are reported in a group incidence table.

Grip strength and sensory reactivity to stimuli

Once during the study, towards the end of treatment, 5 males and 5 females were randomly selected from each group for evaluation of sensory reactivity to stimuli of different modalities (e.g. auditory, visual and proprioceptive stimuli); an assessment of grip strength was also performed. Measurements were performed using a computer generated random order. For males the tests were performed on day before necropsy of the study and for females on Day 3 post partum.

Motor activity assessment (MA)

Once during the study, towards the end of treatment, 5 males and 5 females were randomly selected from each group and the motor activity was measured (for approximately 5 minutes) by an automated activity recording device. Measurements were performed using a computer generated random order. For males the tests were performed the day before necropsy and for females on Day 3 post partum.

Body weight

Males were weighed weekly from allocation to termination.

Females were weighed weekly from allocation to positive identification of mating and on Days 0, 7, 14 and 20 post coitum. Dams were also weighed on Days 1 and 4 post partum.

Food consumption

The weight of food consumed by each cage of males and females was recorded weekly during the pre-mating period starting from allocation.
Individual food consumption for the females was measured on gestation Days 7, 14 and 20 post coitum starting from Day 0 post coitum and on Day 4 post partum starting from Day 1 post partum.

Litter examination
A parturition check was performed from Day 20 to Day 25 post coitum. Females which did not give birth after 25 days of post coitum period were sacrificed shortly after. Gestation length was calculated as the time between the day of successful mating (Day 0 post coitum) and the day of commencement of birth (i.e. first detected presence of offspring in the cage). The day that offspring were first detected in the cage was considered Day 0 post partum.

As soon as possible, after parturition was considered complete (Days 0 or 1 post partum), all pups (live and dead) were counted, sexed and live pups were identified. Live pups were individually weighed on Days 1 and 4 post partum.
Pups killed or dying during the lactation period were weighed before the despatch to necropsy.
Observation was performed once daily for all litters.

Pups
All pups found dead in the cage were examined for external and internal abnormalities.
All live pups sacrificed at termination were examined for external abnormalities and sex confirmation by gonadal inspection. All pups with
abnormalities were retained in an appropriate fixative.

Sacrifice and pathology:

One parental animal killed for humane reasons (no. 90920061) and those that had completed the scheduled test period were killed by exsanguination
under isofluorane anaesthesia.

Parental males:
The males were killed after the mating of all females, after 32 days of treatment period.

Parental females:
The females with live pups were killed on Day 4 post partum.
The females which did not give birth were sacrificed 26/27 days after positive identification of mating.

The clinical history of the males and females of the parental generation was studied and a detailed post mortem examination was conducted (including
examination of the external surface and orifices).
Changes were noted, the requisite organs weighed (excluding animals sacrificed for humane reasons or found dead) and the required tissue samples
preserved in fixative and processed for histopathological examination.

Females:
All females were examined also for the following:

a) external and internal abnormalities;
b) number of visible implantation sites (pregnant animals);
c) number of corpora lutea (pregnant animals).

Organ weights

From all animals completing the scheduled test period, the organs were dissected free of fat and weighed.
The ratios of organ weight to body weight were calculated for each animal.

Tissues fixed and preserved

Samples of all the tissues were fixed and preserved in 10% neutral buffered formalin (except eyes, optic nerves and Harderian glands, testes and
epididymides which were fixed in modified Davidson's fluid and preserved in 70% ethyl alcohol).

Histopathological examination

The tissues required for histopathological examination are listed in Annex 1. After dehydration and embedding in paraffin wax, sections of the tissues
were cut at 5 micrometer thickness and stained with haematoxylin and eosin.
In addition, the testes and epididymides were cut at 2-3 micrometer thickness and stained with Periodic Acid Schiff (PAS). The morphological evaluation of
the seminiferous epithelium (staging of spermatogenic cycle) was performed.

The examination was restricted as detailed below:

a) Tissues specified in sAnnex 1 from 5 males and 5 females randomly selected (animals evaluated for clinical pathology) in the control and high dose
groups killed at term.
b) Tissues specified in Annex 1 from all animals killed or dying during the treatment period.
c) All abnormalities in all groups.

Statistics:

Standard deviations were calculated as appropriate. For continuous variables the significance of the differences amongst group means was assessed by Dunnett’s test or a modified t test, depending on the homogeneity of data.
Statistical analysis of histopathological findings was carried out by means of the non-parametric Kolmogorov-Smirnov test if n was more than 5. The
non-parametric Kruskal-Wallis analysis of variance was used for the other parameters. Intergroup differences between the control and treated
groups were assessed by the non-parametric version of the Williams test. The criterion for statistical significance was p<0.05. The mean values,
standard deviations and statistical analysis were calculated from actual values in the computer without rounding off.

Clinical signs:

no effects observed

Description (incidence and severity):

Statistically significant increase of glucose (50%) and potassium (14%) and decrease of cholesterol (27%), protein (9%), albumin (11%), calcium (3%)
and sodium (1%) were recorded in males dosed with 1000 mg/kg/day. No changes were recorded in treated females. Due to the slight severity of the
above changes, the recorded findings were not considered adverse.

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Description (incidence and severity):

Means of body weight and body weight gain were comparable between control and treated groups both in males and females throughout the study.
The statistically significant decrease in body weight and/or body weight gain, detected in low dose group of males before pairing and during mating, as
well as the statistically significant increase in body weight gain of low dose group of females before pairing were considered of no toxicological relevance
since the changes were minimal and not dose-related.
In addition, decreases in body weight and body weight gain, noted during the post partum phase in control and treated females, were considered as a
normal consequence of the parturition occurred in the females.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

Food consumption was unaffected by treatment in both sexes during the study. The statistically decrease detected in the high dose group on Day 7
post coitum was minimal and not considered related to treatment.

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Behaviour (functional findings):

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Neuropathological findings:

no effects observed

Description (incidence and severity):

Functional Observation BatteryTests: Observation of animals at removal from the cage and in an open arena (neurotoxicity assessment) did not reveal changes attributable to the test item.
Motor activity and sensory reaction to stimuli: No relevant differences were noted in all parameters investigated between control and treated groups.

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Details on results:

Parenteral animals
Mortality

No mortality occurred throughout the study. A total of 3 females were found not pregnant at necropsy: 2 in the mid-dose group and 1 in the high
dose group. The number of females with live pups on Day 4 post partum was: 10 each in the control and low dose groups (0 and 100 mg/kg bw/day), 8 in the mid-dose group (300 mg/kg bw/day) and 9 in the high dose group (1000 mg/kg bw/day).

Clinical signs and clinical observations (Functional Observation Battery Tests)

No relevant clinical signs were observed throughout the study in all treated animals of both sexes.
Observation of animals at removal from the cage and in an open arena (neurotoxicity assessment) did not reveal changes attributable to the test
item.

Motor activity and sensory reaction to stimuli

No relevant differences were noted in all parameters investigated between control and treated groups

Body weight and body weight gain
Means of body weight and body weight gain were comparable between control and treated groups both in males and females throughout the study.
The statistically significant decrease in body weight and/or body weight gain, detected in low dose group of males before pairing and during
mating, as well as the statistically significant increase in body weight gain of low dose group of females before pairing were considered of no
toxicological relevance since the changes were minimal and not dose-related.
In addition, decreases in body weight and body weight gain, noted during the post partum phase in control and treated females, were considered as a normal consequence of the parturition occurred in the females.

Food consumption

Food consumption was unaffected by treatment in both sexes during the study. The statistically decrease detected in the high dose group on Day 7
post coitum was minimal and not considered related to treatment.

Haematology

Reticulocytes were decreased in animals dosed with 1000 mg/kg/day (approximately 33%). Due to the absence of changes in the red cells
parameters, the above reticulopenia was considered toxicologically irrelevant. In addition, the statistically significant decrease of mean
corpuscular haemoglobin concentration, recorded in males dosed with 100 mg/kg/day and 1000 mg/kg/day (2%), was considered of no
toxicological importance due to its minimal severity. No changes were recorded in coagulation parameters.

Clinical chemistry

Statistically significant increase of glucose (50%) and potassium (14%) and decrease of cholesterol (27%), protein (9%), albumin (11%), calcium (3%)
and sodium (1%) were recorded in males dosed with 1000 mg/kg/day. No changes were recorded in treated females. Due to the slight severity of
the above changes, the recorded findings were not considered adverse.


Oestrus cycle, reproductive parameters, pairing combination and mating performance

Oestrous cycle and reproductive parameters (pre-coital intervals, copulatory and fertility indices) did not show relevant differences between treated and control groups.

Implantation, pre-birth loss data and gestation length of females

Corpora lutea, implantations, pre-implantation loss and total litter size were similar between treated and control groups. The slight increase in
pre-birth loss percentage detected in mid- and high dose groups when compared to controls was not statistically significant and not dose-related. Gestation periods were similar in treated groups and controls.

Terminal body weight and organ weights

Terminal body weight was unaffected by treatment in both sexes. Some statistically significant differences were noted in the absolute and/or
relative organ weight of treated animals when compared to controls, such as:
– Higher absolute heart weight in low dose females (+11%).
– Lower absolute spleen weight in high dose females (-19%).
– Higher absolute and relative thymus weight in mid-dose females (+35% and +37% respectively).
– Higher relative kidneys weight in low dose males (+ 9%) and high dose females (13%).
All the above differences were of low magnitude and occurred without dosedependency. Therefore, they were considered unrelated to treatment.

Macroscopic observations
Detailed macroscopic observations have been reported for male and female animals from all groups. No remarkable changes were noted at post
mortem examination in treated animals when compared to controls.

Microscopic observations

Histopathological evaluation was performed on five randomly selected control and high dose males and females, as well as on all abnormalities
detected during post mortem observation. In addition, a detailed qualitative examination of the testes was performed in five randomly selected
control and high dose group males. No treatment-related findings were observed in high dose males and females. Seminiferous tubules were
evaluated with respect to their stage in the spermatogenic cycle and to the integrity of the various cell types within the different stages; regular
layering in the germinal epithelium was noted. The lesions reported in control and treated animals were considered to be an expression of
spontaneous and/or incidental pathology, commonly seen in this species and age under our experimental conditions.

Results of Offspring
Litter data at birth, on Day 1 and on Day 4 post partum of females and sex ratio of pups

No differences in total and live litter size, litter weight, mean pup weight and sex ratio were noted between groups at birth and on Day 4 post partum.

Clinical signs of pups

Apparently no food intake (milk), small appearance and pallor were the clinical signs noted in pups of the control and treated groups. A
malformation (acaudia) was detected in one foetus (female no. 96380039) of the low dose group. These clinical signs noted in pups were
considered unrelated to treatment.

Necropsy findings in decedent pups and in pups sacrificed on Day 4 post partum

Necropsy findings in decedent pups and in pups sacrificed on Day 4 post partum did not reveal any treatment-related effect.


Conclusions

On the basis of the results obtained in this study, the NOAEL (No Observed Adverse Effect Level) for general toxicity was considered to be 1000 mg/kg/day for males and females.

Key result

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: general toxicity

Critical effects observed:

not specified

Conclusions:

On the basis of the results obtained in this study with Butyl diglycol methacrylate, the NOAEL (No Observed Adverse Effect Level) the NOAEL for general toxicity was found to be 1000 mg/kg/day for males and females.

Executive summary:

In a Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test according to OECD Guideline 422 (22 March 1996) Butyl diglycol methacrylate was administered to 10 Hsd: Sprague Dawley SD rats/sex/dose orally by gavage at dose levels of 0 (control), 100, 300 and 1000 mg/kg bw/d. The treatment schedule included 2 weeks before pairing for males and during pairing with females until the day before necropsy, for a total of 29/30 days. Females were treated for 2 weeks prior to pairing, during pairing and throughout the gestation and lactation periods until Day 3 post partum. The following investigations were performed in all groups: body weight, clinical signs (including neurotoxicity assessment, motor activity and sensory reaction to stimuli), food consumption, oestrous cycle, mating performance, clinical

pathology investigations (haematology and clinical chemistry), litter data, macroscopic observations, organ weights and histopathological examination. Clinical signs and macroscopic observations of pups were also performed. The histopathological examination was performed only on control and high dose groups (five animals/sex/group selected randomly). It included identification of the stages of the spermatogenic cycle in five males.

No animals died during the study. A total of 3 females were found not pregnant at necropsy: 2 in the mid-dose group and 1 in the high dose group. The number of females with live pups on Day 4 post partum was: 10 each in the control ad low dose groups, 8 in the mid-dose and 9 in the high dose groups.

No relevant clinical signs were seen throughout the whole study in treated animals of both sexes.

Clinical observations for neurotoxicity assessment (removal of animals from the home cage and open arena) did not reveal changes attributable to the test item.

Motor activity and sensory reaction to stimuli revealed in no relevant differences in all parameters investigated between control and treated groups of both sexes.

No differences of toxicological significance in body weight and body weight gain were recorded in animals of both sexes compared to the control group, throughout the study.

Food consumption was unaffected by treatment in both sexes during the study.

Haematology: No changes of toxicological relevance were seen.

Clinical chemistry: No adverse findings were detected at any dose.

All females mated both in the control and treated groups. Oestrous cycle, precoital intervals, copulatory index and fertility index did not show intergroup differences.

Corpora lutea, implantations, pre-implantation and pre-birth loss percentages, total litter size and gestation length did not reveal any treatment-related effect. No differences in total and live litter size, litter weight, mean pup weight and sex ratio were noted between groups at birth and on Day 4 post partum.

Clinical signs noted in pups throughout the study were considered unrelated to treatment.

Necropsy findings in decedent pups and in pups sacrificed on Day 4 post partum did not reveal any treatment-related effect.

Terminal body weight was unaffected by treatment in both sexes. Changes in organ weights were of slight severity and therefore considered of no toxicological significance.

Macroscopic observations: No remarkable changes were noted at post mortem examination in treated animals when compared to controls.

Microscopic observations: No treatment-related findings were observed in high dose males and females.

Conclusions:

This study is acceptable and satisfies the guideline requirement for a Screening study according to OECD 422 in rats.

On the basis of the results obtained in this study, the NOAEL (No Observed Adverse Effect Level) for general toxicity was considered to be 1000 mg/kg/day for males and females.

NOTE: Any of data in this dataset are disseminated by the European Union on a right-to-know basis and this is not a publication in the same sense as a book or an article in a journal. The right of ownership in any part of this information is reserved by the data owner(s). The use of this information for any other, e.g. commercial purpose is strictly reserved to the data owners and those persons or legal entities having paid the respective access fee for the intended purpose.

 

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

Relaible witout restriction (Klimisch score: 1)

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - systemic effects

Link to relevant study records

Reference

Endpoint:

sub-chronic toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1993

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Remarks:

minor deviations to OECD guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 411 (Subchronic Dermal Toxicity: 90-Day Study)

Deviations:

yes

Remarks:

minor deviations to OECD guideline

GLP compliance:

not specified

Limit test:

no

Specific details on test material used for the study:

- Name of test material (as cited in study report): diethylene glycol butyl ether (DGBE)
- Supplier: Dow Chemical Company
- Physical state: liquid
- Stability under test conditions: yes

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: at room temperature under nitrogen

Species:

rat

Strain:

Sprague-Dawley

Details on species / strain selection:

CD rats

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: CharlesRiver Lab., Raleigh,NC (USA)
- Age at study initiation: 8 weeks

- Housing: individually in suspended stainless steel cages
- Diet: Certified Purina Rodent Chow mash diet ad libitum
- Water: from Elizabethtown Water Company, ad libitum
- Acclimation period: 2 weeks

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24 (67 - 76 °F)
- Humidity (%): 30 - 70 %

- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

occlusive

Vehicle:

water

Remarks:

distilled

Details on exposure:

TEST SITE
- Area of exposure: back (clipped skin)
- % coverage: 3 x 3 cm area
- Type of wrap if used: polyethylene patch, covered by an adhesive banage wrapped around the trunk


REMOVAL OF TEST SUBSTANCE
- Washing (if done): residual test material was gently wiped from the application site
- Time after start of exposure: 6 hrs

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 200, 600 and 2000 mg/kg/d
- Concentration (if solution): undiluted, 10 % solution and 30 % solution
- Constant volume or concentration used: yes

Analytical verification of doses or concentrations:

yes

Duration of treatment / exposure:

13 weeks

Frequency of treatment:

5 days / week , 6 hrs daily

Dose / conc.:

0 mg/kg bw/day

Remarks:

untreated control

Dose / conc.:

200 mg/kg bw/day (nominal)

Remarks:

equals 10% solution

Dose / conc.:

600 mg/kg bw/day (nominal)

Remarks:

equals 30% solution

Dose / conc.:

2 000 mg/kg bw/day (nominal)

Remarks:

undiluted equals 100% of the test sustance

No. of animals per sex per dose:

10

Control animals:

yes

Details on study design:

- Post-exposure recovery period in satellite groups: no

Positive control:

no

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes (observation twice daily for morbidity/rnortality and overt signs of toxic effects)
- Time schedule: twice daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule for examinations: each treatment day (5 days/week) prior to the application of the test article and after the wrappings were removed

DERMAL IRRITATION (if dermal study): Yes
- Time schedule for examinations: each treatment day (5 days/week) prior to the application of the test article and after the wrappings were removed

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION: Yes
- Time schedule for examinations: weekly


OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: pre-study and near the end of the study

HAEMATOLOGY: Yes
- Time schedule for collection of blood: pre-study, after 1 month, and at the end of the study
- Anaesthetic used for blood collection: Yes (light)
- Animals fasted: Yes
- How many animals: all toxicity study animals


CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: pre-study, after 1 month, and at the end of the study
- Animals fasted: Yes
- How many animals: all toxicity study animals


URINALYSIS: Yes
- Time schedule for collection of urine: pre-study, after 1 month, and at the end of the study
- How many animals: all toxicity study animals
- Metabolism cages used for collection of urine: No data
- Animals fasted: Yes / No / No data
- Parameters checked in table [No.?] were examined.

NEUROBEHAVIOURAL EXAMINATION: Yes / No / No data
- Time schedule for examinations:
- Dose groups that were examined:
- Battery of functions tested: sensory activity / grip strength / motor activity / other:

OTHER:

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes, incl. adrenal glands, kidneys, pituitary gland, prostate gland, testes, epididymides, seminal vesicles, liver, ovaries, spleen;
multiple sections of testes, epididymides and ovaries were prepared for examination

Other examinations:

During a 14-day period near the end of the study, daily vaginal smears were performed to determine whether normal estrous cycling was occurring.

Statistics:

Bartlett's test / ANOVA (parametric procedures)
Kruskal-Wallis test (non-parametric procedures)
Furtherly used: Dunn's summed-rank test; Jonckheere's test for monotonic trends for nonparametric comparisons; Fisher exact test.

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

Clinical observations during the toxicity study revealed one mid-dose and one high-dose female with hematuria or red urinary staining on the haircoat, first seen at week 7 of the study. Urinalyses at the end of the study revealed
a slightly increased incidence of occult blood in the urine of females treated with 30 or 100% DGBE(Table I). However, microscopic examination of the urine revealed no urinary casts and no significant numbers of erythrocytes.
see Urinalysis

Dermal irritation:

effects observed, treatment-related

Description (incidence and severity):

Evaluation of the application sites revealed dermal irritation, which was concentration-dependent in incidence, severity, and time of onset and was slightly more severe in females than in males.
Weekly evaluations and dermal scoring performed immediately after removal of the wrappings generally indicated more irritation than when the evaluations were made daily just prior to dosing, which indicated that some recovery occurred overnight
from any acute irritation produced during the daily dosing. Application of undiluted DGBE produced dermal irritation in all animals; some were affected after as few as two doses.
Males generally exhibited very slight or slight erythema with some desquanation or atonia, while most of the females exhibited more severe dermal effects such as areas of necrosis, generally with eschar formation
and subsequent sloughing of the scab at one or more intervals during the study. The necrosis and scab formation were located in most cases at the margin of the wrappings, suggesting that there may have been some mechanical
abrasion due to the wrapping procedure.
Microscopic examination of skin sections from the application site revealed no DGBE-related histological changes, slight thickening of the epidermis being seen for both control and treated animals.

Mortality:

no mortality observed

Description (incidence):

There was no mortality in any of the groups throughout the toxicity or the reproductive phases of the study.

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

not specified

Water consumption and compound intake (if drinking water study):

not specified

Ophthalmological findings:

no effects observed

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Endocrine findings:

not examined

Urinalysis findings:

effects observed, treatment-related

Description (incidence and severity):

slighlty increased incidence of occult urinary blood in mid-and high dose females

Behaviour (functional findings):

not specified

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Details on results:

Female animals were significantly more sensitive to skin irritation

Key result

Dose descriptor:

LOAEL

Effect level:

200 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

dermal irritation

Key result

Dose descriptor:

NOAEL

Effect level:

2 000 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Systemic toxicity

Critical effects observed:

not specified

Conclusions:

No evidence for systemic toxicity in male and female rats; Skin irritation dependent on time, concentration and sex (pronounced effects in females).

Executive summary:

The alcohol Butyldigylycol (CAS 112-34-5) was tested for subchronic toxicity in a well conducted 90 d study in male and female rats (10 animals per sex and dose group). The substance was applied dermally at doses of 200, 600 and 2000 mg/kg/day (6 -hour exposure interval, 5 days/week for 13 weeks). Dermal irritation occurred which was dependent on time and concentration. Female animals were significantly more sensitive. There was a slightly increased incidence of occult blood in the urine of 2 femals (mid-and high dose) which showed no clear time or dose dependency and was not confirmed by microscopic examination of the urine. There was no evidence for systemic toxicity by clinical and histopathological evaluations for all dose groups. Therefore, the NOAEL of the study can be determined to 2000 mg/kg/d.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

2 000 mg/kg bw/day

Study duration:

subchronic

Species:

rat

Quality of whole database:

Study well documented and in accordance to scientifically accepted principles, minor deviations to OECD guideline

Repeated dose toxicity: dermal - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

There is one relevant and adequate study available for BDGMA on repeated dose toxicity (RL=1, guideline study according to OECD 422, GLP).

In a Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test according to OECD Guideline 422 (22 March 1996) Butyldiglycol methacrylate was administered to 10 Hsd: Sprague Dawley SD rats/sex/dose orally by gavage at dose levels of 0 (control), 100, 300 and 1000 mg/kg bw/d. The treatment schedule included 2 weeks before pairing for males and during pairing with females until the day before necropsy, for a total of 29/30 days. Females were treated for 2 weeks prior to pairing, during pairing and throughout the gestation and lactation periods until day 3 post partum. The following investigations were performed in all groups: body weight, clinical signs (including neurotoxicity assessment, motor activity and sensory reaction to stimuli), food consumption, oestrous cycle, mating performance, clinical pathology investigations (haematology and clinical chemistry), litter data, macroscopic observations, organ weights and histopathological examination. Clinical signs and macroscopic observations of pups were also performed. The histopathological examination was performed only on control and high dose groups (five animals/sex/group selected randomly). It included identification of the stages of the spermatogenic cycle in five males.

No animals died during the study. A total of 3 females were found not pregnant at necropsy: 2 in the mid-dose group and 1 in the high dose group. The number of females with live pups on Day 4 post partum was: 10 each in the control ad low dose groups, 8 in the mid-dose and 9 in the high dose groups.

No relevant clinical signs were seen throughout the whole study in treated animals of both sexes.

Clinical observations for neurotoxicity assessment (removal of animals from the home cage and open arena) did not reveal changes attributable to the test item.

Motor activity and sensory reaction to stimuli revealed in no relevant differences in all parameters investigated between

control and treated groups of both sexes.

No differences of toxicological significance in body weight and body weight gain were recorded in animals of both sexes compared to the control group, throughout the study.

Food consumption was unaffected by treatment in both sexes during the study.

Haematology: No changes of toxicological relevance were seen.

Clinical chemistry: No adverse findings were detected at any dose.

All females mated both in the control and treated groups. Oestrous cycle, precoital intervals, copulatory index and fertility index did not show intergroup differences.

Corpora lutea, implantations, pre-implantation and pre-birth loss percentages, total litter size and gestation length did not reveal any treatment-related effect. No differences in total and live litter size, litter weight, mean pup weight and

sex ratio were noted between groups at birth and on Day 4 post partum.

Clinical signs noted in pups throughout the study were considered unrelated to treatment.

Necropsy findings in decedent pups and in pups sacrificed on Day 4 post partum did not reveal any treatment-related effect.

Terminal body weight was unaffected by treatment in both sexes. Changes in organ weights were of slight severity and therefore considered of no toxicological significance.

Macroscopic observations: No remarkable changes were noted at post mortem examination in treated animals when compared to controls.

Microscopic observations: No treatment-related findings were observed in high dose males and females.

The alcohol Butyldigylycol (CAS 112-34-5) was tested for subchronic toxicity in a well conducted 90 d study in male and female rats (10 animals per sex and dose group). The substance was applied dermally at doses of 200, 600 and 2000 mg/kg. Dermal irritation occurred which was dependent on time and concentration. Female animals were significantly more sensitive. There was a slightly increased incidence of occult blood in the urine of 2 femals (mid-and high dose) which showed no clear time or dose dependency and was not confirmed by microscopic examination of the urine. There was no evidence for systemic toxicity by clinical and histopathological evaluations for all dose groups. Therefore, the NOAEL for systemic toxicity can be determined to 2000 mg/kg/d.

Conclusions

On the basis of the results obtained in the OECD TG 422 study on BDGMA, the NOAEL (No Observed Adverse Effect Level) for general toxicity was considered to be 1000 mg/kg/day for males and females.

Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:
OECD guideline 422 study, Klimisch score: 1, no deviations, GLP.

Justification for selection of repeated dose toxicity dermal - systemic effects endpoint:
Pivotal metabolite, relevant pathway of exposure

Justification for classification or non-classification

Based on the available data, BDGMA does not need to be classified for repeated dose toxicity according to the criteria given in regulation (EC) 1272/2008 or the former European directive on classification and labelling 67/548/EEC. Thus, no labelling is required.