Amines, C12-14-alkyl, C6-10-alkyl phosphates

Repeated dose toxicity: oral

Administrative data

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

2013

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.7 (Repeated Dose (28 Days) Toxicity (Oral))

Test guideline 3

Qualifier:

according to guideline

Guideline:

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

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: OECD Guideline 421

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Amines, C11-14-branched alkyl, monohexyl and dihexyl phosphates

EC Number:

279-632-6

EC Name:

Amines, C11-14-branched alkyl, monohexyl and dihexyl phosphates

Cas Number:

80939-62-4

Molecular formula:

na (UVCB)

IUPAC Name:

Amines, C11-14-branched alkyl, monohexyl and dihexyl phosphates

Test material form:

liquid: viscous

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Administration / exposure

Route of administration:

oral: gavage

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Accuracy, homogeneity and stability of formulations were demonstrated by analyses.

Duration of treatment / exposure:

Males - 29 d and females - 41-54 d

Frequency of treatment:

Once daily

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

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

Remarks:

Based on the results of a 14-day dose range finding study (Project 499977; BASF Project 01R0613/11X493) the dose levels for this combined 28-day oral gavage study with reproduction/developmental toxicity screening test were selected to be 10, 30 and 100 mg/kg bw/day.

Doses / concentrations 2

Dose / conc.:

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

Doses / concentrations 3

Dose / conc.:

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

No. of animals per sex per dose:

10

Control animals:

yes, concurrent vehicle

Details on study design:

After acclimatization, four groups of ten male and ten female Wistar Han rats were exposed by oral gavage to the test substance at 0, 10, 30 and 100 mg/kg/day. Males were exposed for 29 days, i.e. 2 weeks prior to mating, during mating, and up to termination. Females were exposed for 41 - 54 days, i.e. during 2 weeks prior to mating, during mating, during post-coitum, and during at least 4 days of lactation.

Examinations

Observations and examinations performed and frequency:

The following observations and examinations were evaluated: mortality / viability, clinical signs (daily), functional observations and locomotor activity (Week 4 (males); end of lactation (females)), body weight and food consumption (at least at weekly intervals), clinical pathology (Week 4 (males); end of lactation (females)), macroscopy at termination, organ weights and histopathology on a selection of tissues.

Results and discussion

Results of examinations

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Haematological findings:

effects observed, treatment-related

Clinical biochemistry findings:

no effects observed

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Gross pathological findings:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Details on results:

Hematological examinations revealed slightly increased leucocyte counts in males and females at 100 mg/kg bw/day, primarily due to increased neutrophil counts. Increased leucocyte counts were also present in females at 30 mg/kg bw/day, and the percentage of neutrophils was increased in both sexes at this dose. No toxicologically relevant changes were observed by clinical chemistry examinations. Spleen weights were moderately increased in both sexes at 100 mg/kg bw/day.
Enlarged spleen was observed in one male and 4 females at 100 mg/kg bw/day. Enlarged mesenteric lymph nodes were present in 5 males and all females at high dose, in 6 females at the mid dose and in 2 females at the low dose. Treatment-related microscopic findings were noted in the mesenteric lymph nodes and adrenal gland cortex (all dose groups). Findings in the mesenteric lymph nodes consisted of:
− foamy macrophages with/without fibrosis (up to a marked degree), which was correlated with enlargement of the mesenteric lymph nodes at necropsy.
− Sinus ectasia (up to a moderate degree) at all doses, which was considered secondary to the occurrence of foamy macrophages.
− Focal necrosis within the aggregates of foamy macrophages with/without fibrosis of males at all doses and in females at the mid and high dose
− Macrophage foci up to a moderate degree (females) or marked degree (males) at all doses.
The lesions in the adrenal cortex consisted of inflammatory lymphocytic cells (up to a slight degree) in females at all doses.

Effect levels

Key result

Dose descriptor:

LOAEL

Effect level:

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

Based on:

test mat.

Sex:

male/female

Basis for effect level:

gross pathology

haematology

histopathology: non-neoplastic

organ weights and organ / body weight ratios

Target system / organ toxicity

Key result

Critical effects observed:

yes

Lowest effective dose / conc.:

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

System:

haematopoietic

Organ:

adrenal glands

mesenteric lymph node

spleen

Treatment related:

yes

Dose response relationship:

yes

Applicant's summary and conclusion

Conclusions:

Under the study conditions, no NOAEL could be determined for parental systemic toxicity and hence the LOAEL for systemic toxicity was determined to be 10 mg/kg bw/day

Executive summary:

A study was conducted to determine the repeated dose toxicity potential of the test substance according to OECD Guideline 422, 421 and 407 and EU Method B.7, under GLP conditions. Four groups of ten male and ten female Wistar Han rats were exposed by oral gavage to the test substance at 0, 10, 30 and 100 mg/kg bw/day. Males were exposed for 29 d, i.e. 2 weeks prior to mating, during mating, and up to termination. Females were exposed for 41 - 54 d, i.e. during 2 weeks prior to mating, during mating, during post-coitum, and during at least 4 d of lactation. The following observations and examinations were evaluated: mortality / viability, clinical signs (daily), functional observations and locomotor activity (Week 4 (males); end of lactation (females)), body weight and food consumption (at least at weekly intervals), clinical pathology (Week 4 (males); end of lactation (females)), macroscopy at termination, organ weights and histopathology on a selection of tissues, and reproduction/developmental parameters, consisting of mating, fertility and conception indices, precoital time, number of corpora lutea and implantation sites, gestation index and duration, parturition, maternal care, sex ratio and early postnatal pup development (mortality, clinical signs, body weights and macroscopy). Formulations were analyzed once during the study to assess accuracy, homogeneity and stability. No clinical signs of toxicity were observed in the parental animals. Body weight development and food consumption were not affected by administration of the test substance. Hematological examinations revealed slightly increased leucocyte counts in males and females at 100 mg/kg bw/day, primarily due to increased neutrophil counts. Increased leucocyte counts were also present in females at the mid dose and the percentage of neutrophils was increased in both sexes at this dose. No toxicologically relevant changes were observed by clinical chemistry examinations. Enlarged spleen with increased weight was observed at the high dose only. Histopathological findings were confined to lesions in the mesenteric lymph nodes (foamy macrophages with/without fibrosis, sinus ectasia, focal necrosis within the aggregates of foamy macrophages with/without fibrosis and macrophage foci) and adrenal gland cortex (inflammatory lymphocytic cells). The higher neutrophil counts (and white blood cell counts) at 30 and 100 mg/kg bw/day were considered to be due to these inflammatory histopathological changes. No treatment-related changes were noted in any of the reproductive parameters investigated in this study. Histopathologically, the assessment of the integrity of the spermatogenetic cycle did not provide any evidence of impaired spermatogenesis, and no abnormalities were seen in the reproductive organs of the animal that failed to deliver healthy offspring. Finally, no treatment-related changes were noted in any of the developmental parameters investigated in this study. Under the study conditions, no NOAEL could be determined for parental systemic toxicity and hence the LOAEL for systemic toxicity was determined to be 10 mg/kg bw/day (van Otterdijk, 2013).