Alcohols, C16-18, ethoxylated, phosphates

Toxicity to reproduction

Administrative data

Endpoint:

screening for reproductive / developmental toxicity

Remarks:

based on test type (migrated information)

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 days (females)
- Weight at study initiation: 306-344 (males), 208-239 (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 days

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 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 ot 60 minutes before dosage administration.

Details on mating procedure:

- M/F ratio per cage: 1/1
- Length of cohabitation: maximum of 14 days
- Proof of pregnancy: sperm in vaginal smear and/or copulatory plug referred to as day 0 of pregnancy
- Further matings after two unsuccessful attempts: no
- After successful mating each pregnant female was caged (how): individually

Analytical verification of doses or concentrations:

no

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. No analytical method was developped for this test substance.

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 4 of lactation for dams that delivered a litter, Day 24 of gestation for rats that did not deliver a litter, study Day 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. 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

Parental animals: Observations and examinations:

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 Days 0, 7, 10, 12, 15, 18, 20, 24, and 25 of gestation, and on Day 1 and 4 of lactation. For the rat with no confirmed mating on study Days 28, 35, 38, 40, 43, 46, 48, and 52.

Oestrous cyclicity (parental animals):

Estrous cycling was evaluated by examination of vaginal cytology for 13 days beginning with the day after the first administration and then until spermatozoa were observed in a smear of the vaginal contents and/or a copulatory plug was observed in situ during the cohabitation period.

Sperm parameters (parental animals):

Parameters examined in the male parental generation: testis weight, epididymis weight

Litter observations:

PARAMETERS EXAMINED
The following parameters were examined in [F1] offspring:
number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, weight gain, physical or behavioural abnormalities

GROSS EXAMINATION OF DEAD PUPS:
yes, for external and internal abnormalities; possible cause of death was determined for pups born or found dead

Postmortem examinations (parental animals):

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

Postmortem examinations (offspring):

SACRIFICE
- The F1 offspring were sacrificed at Day of lactation 4 or 5
- These animals were subjected to postmortem examinations (macroscopic and/or microscopic examination) as follows:
examinaton for gross lesions and an internal confirmation of sex.

GROSS NECROPSY
- Gross necropsy included a single cross-section of the head at the level of the frontal-parietal suture and examination of the cross-sectioned brain for apparent hydrocephaly. Pups that died before examination of the litter for pup viability were evaluated for vital status at birth. The lungs were removed and immersed in water. Pups with lungs that sank were identified as stillborn; pups with lungs that floated were identified as liveborn, and to have dies shortly after birth.

Reproductive indices:

- Fertility index = (number of pregnant animals / number of copulated pairs) x 100
- Gestation index = (number of pregnant females with pups alive / number of pregnant females) x 100

Offspring viability indices:

- Viability index = (number of pups alive on day 4 / number of pups alive on day 0) x 100

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

800 mg/kg bw/d: excess salivation in both sexes

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

800 mg/kg bw/d: reduced mean bw gain and reduced food consumption (males: week 1-2; females: during gestation)

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

800 mg/kg bw/d: reduced mean bw gain and reduced food consumption (males: week 1-2; females: during gestation)

Organ weight findings including organ / body weight ratios:

no effects observed

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)

Other effects:

no effects observed

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:

no effects observed

Reproductive function: sperm measures:

not examined

Reproductive performance:

no effects observed

Details on results (P0)

CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
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 rat was 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 FOOD CONSUMPTION (PARENTAL ANIMALS)
800 mg/kg bw:
Males: reduction of body weight gain (Day 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 in the 800 mg/kg bw group; lactation: no effects

REPRODUCTIVE FUNCTION: ESTROUS CYCLE (PARENTAL ANIMALS)
No effects observed.

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)
Pregnancy occurred in 90%, 100%, 90%, 90%, and 80% female rats in the control, 25, 50, 200, and 800 mg/kg bw dose groups, respectively. Two pregnant females in the 800 mg/kg bw group were found dead or sacrificed before delivery. All other pregnant rats delivered a litter.
Values for the numbers of dams delivering litters and the gestation index, the numbers of dams with stillborn pups and of dams with all pups dying, viability and lactation indices, and the sex ratio were comparable within all groups.

ORGAN WEIGHTS (PARENTAL ANIMALS)
No treatment-related effects were observed.

GROSS PATHOLOGY (PARENTAL ANIMALS)
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 (PARENTAL ANIMALS)
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 (PARENTAL ANIMALS)
No clear and consistent treatment related differences in results for the qualitative and quantitative mcroscopic assessment of bone marrow smears were observed.

Effect levels (P0)

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:

fertility

Effect level:

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

Based on:

test mat.

Sex:

male/female

Results: F1 generation

General toxicity (F1)

Clinical signs:

no effects observed

Mortality / viability:

no mortality observed

Body weight and weight changes:

not examined

Sexual maturation:

not examined

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

no effects observed

Histopathological findings:

not examined

Details on results (F1)

VIABILITY (OFFSPRING)
Natural delivery and litter observations were unaffected by dosages.

CLINICAL SIGNS (OFFSPRING)
800 mg/kg bw: Cold to touch, not nesting, moderate dehydration and pale were observed. They were not attributed to the dosage (no dose-response, only one litter, and/or only observed in the control group).

GROSS PATHOLOGY (OFFSPRING)
No effects observed.

OTHER FINDINGS (OFFSPRING)
800 mg/kg bw: Values for the duration of gestation and pup weights per litter were increased (not statistically significant). Implantation sites per litter, corpora lutea, litter sizes, surviving pups per litter and live litter size at weighing were reduced (not statistically significant). These changes were not considered to be biologically relevant.

Overall reproductive toxicity

Reproductive effects observed:

not specified

Any other information on results incl. tables

Table 1: Reproductive/developmental toxicity screening test: parental examinations

	Control group	25 mg/kg bw/day	50 mg/kg bw/day	200 mg/kg bw/day	800 mg/kg bw/day
Body weight (body weight gain) [g]
Males Day 1 (Day 1-8)	336 ± 9 (40 ± 8)	339 ± 9 (35 ± 6)	343 ± 10 (36 ± 9)	340 ± 10 (31 ± 11)	341 ± 10 (16 ± 17**)
Males Day 8 (Day 8-14)	376 ± 13 (31 ± 11)	374 ± 10 (27 ± 6)	379 ± 12 (27 ± 12)	371 ± 17 (27 ± 8)	357 ± 24* (25 ± 11)
Males Day 14	407 ± 21	401 ± 13	406 ± 21	397 ± 22	382 ± 27
Males Day 22 (Day 22-29)	434 ± 28 (23 ± 12)	431 ± 15 (24 ± 5)	430 ± 27 (21 ± 8)	426 ± 29 (21 ± 7)	406 ± 20 (22 ± 7)
Males Day 45 (Day 1-45)	505 ± 54 (169 ± 50)	497 ± 19 (158 ± 16)	491 ± 38 (148 ± 37)	492 ± 41 (152 ± 33)	475 ± 25 (134 ± 21)
Females Day 0 of gestation (Day 0-7)	259 ± 15 (37± 7)	261 ± 10 (37 ± 11)	259 ± 15 (35 ± 5)	261 ± 15 (32 ± 6)	257 ± 22 (17 ± 26**)
Females Day 15 of gestation	339 ± 19	341 ± 21	335 ± 18	329 ± 14	317 ± 30
Females Day 18 of gestation	382 ± 19	379 ± 27	372 ± 18	363 ± 16	336 ± 48**
Females Day 20 of gestation (Day 0-20)	412 ± 23 (153 ± 11)	413 ± 31 (151 ± 23)	397 ± 19 (138 ± 10)	393 ± 17 (132 ± 7)	380 ± 45 (129 ± 25*)

* significantly different from control P<0.05

** significantly different from control P<0.01

 Table 2: Reproductive/developmental toxicity screening test: Reproductive parameters.

	Control group	25 mg/kg bw/day	50 mg/kg bw/day	200 mg/kg bw/day	800 mg/kg bw/day
Mean duration of gestation and overall litter performance values
Fertility index (%)	100	100	90	100	88.9
Number of females delivered a litter	9	10	9	9	6
Gestation index (%)	100	100	100	100	100
Mean number of implant sitesper pregnancy	16± 3	16± 2	15± 2	16± 1	14± 5
Corpora lutea	20± 4	21± 4	21± 5	22± 3	17± 3
Group mean F1 survival indices
Viability Index (%)	98.6	99.3	99.2	100.0	98.6
Lactation index (%)	100	100	100	100	100