Alcohols, C16-18, ethoxylated, phosphates

Repeated dose toxicity: oral

Administrative data

Endpoint:

short-term repeated dose toxicity: oral

Remarks:

combined repeated dose and reproduction / developmental screening

Type of information:

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

Adequacy of study:

key study

Study period:

30 May 2008 - 06 Feb 2009

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

GLP - Guideline study, tested with CAS 68130-47-2. According to the ECHA guidance document “Practical guide 6: How to report read-across and categories (March 2010)”, the reliability was changed from RL1 to RL2 to reflect the fact that this study was conducted on a read-across substance.”

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2009

Report date:

2009

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

68130-47-2

EC Number:

614-291-2

Cas Number:

68130-47-2

IUPAC Name:

68130-47-2

Details on test material:

- Name of test material (as cited in study report): (C8-10) poly(oxy-1,2-ethanediyl), alpha-hydro-omega-hydroxy-, mono-C8-10-alkyl ethers, phosphates
- Substance type: UVCB
- Physical state: clear, yellowish, viscous liquid
- Analytical purity: 100%
- Composition of test material, percentage of components: Linear saturated C8-10 alcohol ethoxylate phosphate with 5-10 moles of ethoxylation
- Lot/batch No.: 1160482
- Stability under test conditions: stable
- Storage condition of test material: room temperature

Test animals

Species:

rat

Strain:

other: Crl:CD(SD)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories, Inc., Portage, MI, USA
- Age at study initiation: 72 d (males), 66 d (females)
- Weight at study initiation: 306-344 g (males), 208-239 g (females)
- Housing: individual housing in stainless steel, wire-bottomed cages.
- Diet: Certified Rodent Diet 5002 (PMI Nutrition International, St.Louis, MO, USA), ad libitum
- Water: filtered local water (chlorine addition), ad libitum
- Acclimation period: 7 d

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-25°C
- Humidity (%): 30-70%
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS:
Formulations were prepared at least once daily. Formulations were used within four hours of preparation and were stirred for at least 30 or 60 minutes before dosage administration.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

A weight/weight verification method was employed to verify the test-substance content of the prepared formulations.

Duration of treatment / exposure:

Males: 14 days before cohabitation period through the day before sacrifice (45 dosages)
Females: 14 days before cohabitation period through the day before scheduled sacrifice (Day of lactation (DL) 4 for dams that delivered a litter, Day of gestation (DG) 24 for rats that did not deliver a litter, study Day (DS) 52 for the rat with no confirmed day of mating). Surviving female rats were given a total of 38 to 52 dosages.

Frequency of treatment:

Once daily

Doses / concentrations

Remarks:

Doses / Concentrations:
25, 50, 200, 800 mg/kg bw
Basis:
actual ingested

No. of animals per sex per dose:

10

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: In a range-finding study rats were dosed with 100, 300, and 1000 mg/kg bw. At day 6 the high dose of 1000 mg/kg bw was lowered to 750 mg/kg bw, due to excessive toxicity. Reductions of the body weight were observed within days 4-5. In the gestation period body weights were only slightly reduced in the 300 and 750/1000 mg/kg bw dose groups. No effects were observed after Caesarean section on Day 21 of gestation up to the highest dose of 750/1000 mg/kg bw. The female animal of the 1000 mg/kg bw that was sacrificed on study Day 4 had received 3 doses. The animal lost a total of 28 g of body weight and consumed an average of 8 g of food per day. Clinical signs including excess salivation, rales, cold/warm to touch, hunched posture, and scant feces were noted. Necropsy revealed two black ulcerations present in the pyloric region of the stomach, dark red lobes of the liver, red lobes of the lungs, and intestines were distended with gas. Thus, doses of 25, 50, 200, and 800 mg/kg bw were chosen for the main study. The low dose was expected to be a NOAEL for maternal and embryo-fetal toxicity, and the 800 mg/kg bw dose was expected to produce minimal maternal toxicity and little or no developmental toxicity.

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily
- Cage side observations: clinical signs, abortions, premature deliveries, deaths

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: before the first dosage and weekly thereafter

BODY WEIGHT: Yes
- Time schedule for examinations: daily

FOOD CONSUMPTION:
- Males: weekly, except for the cohabitation period; females: weekly until the cohabitation period, and on gestation day (DG) 0, 7, 10, 12, 15, 18, 20, DG 24, DG 25, and on days of lactation (DL) 1 and 4. For the rat with no confirmed mating on study days (DS) 28, 35, 38, 40, 43, 46, 48, and 52.

HAEMATOLOGY: Yes
- Time schedule for collection of blood:
- Anaesthetic used for blood collection: No, rats were sacrificed by carbon dioxide asphyxiation.
- Animals fasted: Yes
- How many animals: 4-5
- Parameters checked: erythrocyte count, haematocrit, mean cell volume, mean corpular volume, haemoglobin, leucocyte count, thrombocyte count, mean platelet volume, activated partial thromboplastin time, prothrombine time.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: No, rats were sacrificed by carbon dioxide asphyxiation.
- Animals fasted: Yes
- How many animals: 4-5
- Parameters checked: gamma glutamyl transferase, aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, sodium, potassium, glucose, urea, albumine, globuline, albumin/globulin ratio, protein, calcium, creatinine, cholesterol, chloride, bilirubin, triglycerides.


NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: DS 46 (males), DL 5 and DG 25 (females)
- Dose groups that were examined: 5/sex/group (all dose groups)
- Battery of functions tested: sensory activity / grip strength / motor activity / autonomic functions / abnormal clinical signs

Sacrifice and pathology:

GROSS PATHOLOGY: Yes, whole body

HISTOPATHOLOGY: Yes, small intestine, large intestine, brain, urinary bladder, heart, testes, liver, lung, trachea, lymphnodes, mesentery lymphnodes, spleen, epididymides, adrenal gland, peripheral nerve, kidney, ovary, prostate, vesicula seminalis, thyroid gland, oesophagus, thymus, uterus, bone marrow, mammary gland, spinal cord, sternum, stomach, vagina

Statistics:

Proportion data as well as descriptive and quantal data from the functional observational battery (FOB), was analyzed as contingency tables using the variance test for homogeneity of the binominal distribution.
Bartlett´s test of homogeneity of variances were used to estimate the probability that the dosage groups have different variances. Any non-significant result (p>0.001) indicate that an assumption of homogeneity of variance is anappropriate, and the data were compared using the Analysis of Variance. If p≤0.05, the groups given the test substance were compared with the control group using Dunnett´s Test. If Bartlett´s test was significant (p≤0.001), the Analysis of Variance Test was not appropriate (nonparametric). When 75% or fewer of the scores in all the groups were tied, the Kruskal-Wallis Test was used to analyze the data, and in the event of a significant result (p≤0.05), Dunn´s Test was used to compare the groups given the test substance with the control group. When more than 75% of the scores in any dosage group are tied, Fisher´s Exact Test was used to compare the proportion of ties in the dosage group.
Data from the motor activity test, with repeated measurements within a session, was analyzed using an Analysis of Variance with repeated measures. A significant effect (P≤0.05) in that test could have appeared as effect of Concentration (a difference between groups in the total across all measurements in a session) or as an interaction between Concentration and Block (a difference between groups at specific measurement periods). If the Concentration x Block interaction was significant, an Analysis of Variance Test was used to evaluate the data at each measuremnet period, and a significant result (p≤0.05) was followed by a comparison of the groups using Dunnett´s test. Variables with graded count scores, such as litter size, were analyzed using Kruskal-Wallis (if p≤0.05 followed by Dunn´s Test).

Results and discussion

Results of examinations

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

800 mg/kg bw/d: excess salivation (male and female), chromorhinorrhea, urine-stained abdominal fur, red and/or dried perioral substance and red or clear perinasal substance in males

Mortality:

mortality observed, treatment-related

Description (incidence):

800 mg/kg bw/d: excess salivation (male and female), chromorhinorrhea, urine-stained abdominal fur, red and/or dried perioral substance and red or clear perinasal substance in males

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

800 mg/kg bw/d: reduction of body weight gain

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

800 mg/kg bw/d: reduced food consumption

Food efficiency:

not examined

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

no effects observed

Ophthalmological findings:

no effects observed

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

not examined

Behaviour (functional findings):

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

800 mg/kg bw: damage on the lining of the stomach

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

800 mg/kg bw: findings in the non-glandular stomach related to test substance (8/8 males; 4/4 females)

Histopathological findings: neoplastic:

no effects observed

Details on results:

CLINICAL SIGNS AND MORTALITY
There were no substance-related deaths. One female rat of the 800 mg/kg bw dose group was sacrificed due to adverse clinical observations and two more female rats were found dead due to an intubation error.
In the 800 mg/kg bw group excess salivation, chromorhinorrhea, urine-stained abdominal fur, red and/or dried perioral substance and red or clear perinasal substance occurred in significantly increased numbers of male rats. All other clinical signs were considered unrelated to treatment.
Significantly increased numbers of females in the 800 mg/kg group had excess salivation.

BODY WEIGHT AND WEIGHT GAIN
800 mg/kg bw:
Males: reduction of body weight gain (DSs 1 to 14) and body weight (day 8). Body weight mean on day 45 was 94% of the control value.
Females: Precohabitation: no effects; gestation: reduced body weight gain in the 800 mg/kg bw group; lactation: no effects

FOOD CONSUMPTION:
800 mg/kg bw:
Males: reduction from day 1-14 (absolute and relative) and day 1-43 (relative)
Females:
Precohabitation: reduction from study day (DS) 1-8 (absolute and relative)
Gestation: reduction from days of gestation (DG) 0-7 and 15-18
Lactation: no effects
No effects were observed in the other dose groups.

HAEMATOLOGY
No biologically important differences were observed.
Reductions in APTT for females of the 50 and 800 mg/kg bw group reflects an increased value for the vehicle control. ALT and triglycerides were increased at 50/200 and 25/800 mg/kg bw groups, respectively. These effects were considered unrelated to treatment as no dose dependency was observed.

CLINICAL CHEMISTRY
No biologically important differences were observed.

NEUROBEHAVIOUR
No effects were observed.

ORGAN WEIGHTS
No treatment-related effects were observed.

GROSS PATHOLOGY
800 mg/kg bw:
Males: thickened walls of the cardiac region of the stomach and white areas on the mucosal surface (3/10), ulceration on the mucosal surface of the cardiac region of the stomach (1/10)
Females: lesions of the cardiac region of the stomach (2/10)

HISTOPATHOLOGY: NON-NEOPLASTIC
800 mg/kg bw: findings in the non-glandular stomach related to test substance (8/8 males; 4/4 females)
200 mg/kg bw: findings in the non-glandular stomach possibly related to test substance, irritating effects (1/5 males)
25, 50 mg/kg bw: no adverse effects observed

OTHER FINDINGS
No clear and consistent treatment related differences in results for the qualitative and quantitative mcroscopic assessment of bone marrow smears were observed.

Effect levels

1

Dose descriptor:

NOAEL

Remarks:

systemic

Effect level:

800 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

2

Dose descriptor:

NOAEL

Remarks:

local

Effect level:

200 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

3

Dose descriptor:

LOAEL

Remarks:

local

Effect level:

800 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Irritating effects on the forestomach of the rat. Occurrence of hyperplasia and ulceration.

Target system / organ toxicity

Critical effects observed:

not specified