Reaction mass of ammonium(3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctyl) hydrogen phosphate and ammonium bis(3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctyl) phosphate

Endpoint summary

Description of key information

The oral LD50 for the test substance was greater than 5000 mg/kg for female rats.

The inhalation LD50 for the test substance was between 0.05 and 0.5 mg/L for male rats

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Single dose - 2009

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 425 (Acute Oral Toxicity: Up-and-Down Procedure)

GLP compliance:

yes

Test type:

up-and-down procedure

Species:

rat

Strain:

other: Crl:CD(SD)

Sex:

female

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Control animals:

no

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Interpretation of results:

GHS criteria not met

Conclusions:

A single dose of test substance was administered by oral gavage to 1 fasted female rat each at a dose of 175, 550, or 1750 mg/kg and to 3 fasted female rats at a dose of 5000 mg/kg. The rats were dosed 1 at a time at a minimum of 48 hour intervals. The rats were observed for mortality, body weight effects, and clinical signs for 14 days after dosing. All rats were necropsied to detect grossly observable evidence of organ or tissue damage. No deaths occurred. One of the rats dosed at 5000 mg/kg exhibited high posture on the day of dosing. No other clinical signs were observed. The rats exhibited no body weight losses after dosing. No gross lesions were present in the rats at necropsy. Under the conditions of this study, the oral LD50 for the test substance was greater than 5000 mg/kg for female rats.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

May to October 2019

Reliability:

1 (reliable without restriction)

Qualifier:

according to guideline

Guideline:

OECD Guideline 433 (Acute Inhalation Toxicity: Fixed Concentration Procedure)

GLP compliance:

yes (incl. QA statement)

Test type:

fixed concentration procedure

Limit test:

no

Specific details on test material used for the study:

Batch: W19011506
Purity: [Mono] 2-(perfluorohexyl)ethyl-phosphate: 11.9%
[Bis] 2-(perfluorohexyl)ethyl-phosphate: 19.2%
Expiry: 15 January 2020

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

Animal model Crl:CD (SD) IGS Rats
Supplier Sighting Study : Animals transferred from
in-house Colony (Covance
Study No. 94-0986)
Main Study : Charles River Laboratories
Raleigh, North Carolina 27610
Number of animals Received/Transferred
from the in-house colony
Placed on test
Sighting
Study: Males: 2 Males: 2
Females: 2 Females: 2
Main
Study:
Males: 5 Males: 5
Age of the main study
animals at start of exposure
administration
Approximately 2 months
Weight range of the main
study animals at start of
exposure administration 268.1 g to 280.5

Assignment Animals were allocated into treated animal groups upon transfer
(Sighting Study) or upon arrival (Main Study). For Main Study
animals, individual weights of animals placed on test were
within ± 20% of the mean weight of any previously exposed
animals where practical.
Identification of animals Each main study rat was implanted with a BMDS IMI-500
Implantable Radio Frequency Transponder (microchip)
programmed with a unique number. This number was cross
referenced with an animal number assigned by the Testing
Facility; this number plus the study number comprised a unique
identification number for each animal. In addition, each cage
was provided with a cage card that was color-coded for dose
level identification and contained study number and facilityassigned animal number information.

Light/dark cycle A twelve hour light/dark cycle was provided and controlled via
an automatic timer.
Temperature and relative
humidity
Monitored and maintained within the range of 20 to 26 °C and
30 to 70%.
Although conditions were occasionally outside the indicated
ranges these deviations were minor and/or of short duration and
were not considered to have influenced the health of the animals
and/or the outcome of the study.

Cages Polycarbonate cages with a stainless steel mesh lid.
Number of animals per cage Sighting Study animals were pair housed by sex during the pretest
period and single housed after exposure. Main Study animals
were pair housed or triple housed by sex throughout the study.
Bedding Teklad 7070C Certified Diamond Dry Cellulose Bedding
Envigo, Madison, Wisconsin
Provided to each cage throughout the study and changed at
appropriate intervals each week.
Analytical results of the bedding, provided by the manufacturer,
are maintained on file at the Testing Facility. There were no
known contaminants in the bedding that were expected to interfere
with the objectives of this study.
Environmental enrichment Provided to each cage throughout the study and replaced when
necessary.

Diet Teklad Global 16% Protein Rodent Diet (Certified), 2016C
Envigo, Madison, Wisconsin
Availability Without restriction. Fresh feed was presented weekly
Analysis Analysis of each feed lot used during this study was performed
by the manufacturer. Results were provided to the Testing
Facility and are maintained on file at the Testing Facility. There
were no known contaminants in the feed that were expected to
interfere with the results of this study.

Supply Potable water from the public supply (New Jersey-American
Water Company, Cherry Hill, New Jersey) via an automated
watering system.
Availability Without restriction.
Analysis Water analyses are conducted by New Jersey-American Water
Company, Cherry Hill, New Jersey (Raritan-Millstone Plant) to
ensure that water meets standards specified under the EPA
Federal Safe Drinking Water Act Regulations (40 CFR Part
141). In addition, water samples are collected biannually from
representative rooms in the Testing Facility; chemical and
microbiological water analyses are conducted on these samples
by a subcontract laboratory. Results of all water analyses are
maintained on file at the Testing Facility. There were no known
contaminants in the water which were expected to interfere with
the results of this study.

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

nose only

Vehicle:

water

Mass median aerodynamic diameter (MMAD):

3.1 µm

Geometric standard deviation (GSD):

>= 2.43 - <= 2.91

Details on inhalation exposure:

Route The test item was administered as a liquid aerosol and delivered
to animals by nose-only inhalation for up to 4 hours per day.
Training for dosing The animals on study were acclimated to the method of restraint
once for 30 to 60 minutes preceding their test item exposure.
Duration of daily exposure Up to 240 minutes (4 hours).

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

>= 4 h

Concentrations:

Exposure 1
Target exposure level 0.05 (mg/L/day)
Number of Animals 5

No. of animals per sex per dose:

5 Male animals for main study

Control animals:

no

Details on study design:

Experimental Evaluations
Daily Observations
Animals were observed in their cages twice daily for mortality and general condition. Animals
in extremely poor health or of questionable viability were identified for further monitoring and
possible euthanasia.

Exposure Observations
On exposure days, all animals were observed for signs of toxic effects once prior to and at least
hourly (including upon return to their home cages) up to 4 hours after test item administration.
On non-exposure days animals were observed for signs of toxic effects daily until animals were
euthanized (Sighting Study) or necropsied (Main Study).

Signs
Sighting Study: each animal was removed from its cage and examined twice pretest (including
on the day before exposure) and on Day 7.
Main Study: each animal was removed from its cage and examined once pretest (on the day
before exposure) and on Days 7 and 14.
Examinations included observations of general condition, skin and fur, eyes, nose, oral cavity,
abdomen and external genitalia as well as an evaluation of respiration.

Body Weight
Sighting Study: Each animal was removed from its cage and weighed on Days 1 (prior to
exposure), 2, 4 and 8. Due to adjustment of the Sighting Study exposure schedule, Animal
Nos. 1001M and 1502F received additional body weights.
Main Study: Each animal was removed from its cage and weighed on Days 1 (prior to
exposure), 2, 4, 8 and 15.

Terminal Investigations
Necropsy
Macroscopic Examinations
Complete macroscopic examinations were performed on all unscheduled decedents in the
Sighting Study and on all Main Study animals. The necropsy included an external examination
as well as a detailed internal examination.
Scheduled Necropsy
Surviving Sighting Study animals were euthanized on Day 8 and discarded without examination.
Necropsy was performed on all Main Study animals after animals had been exposed to the test
item on one occasion followed by a 14-day observation period.
Method of Euthanasia
Exsanguination following isoflurane inhalation
Tissues Preserved
Abnormal tissues from unscheduled decedents from the Sighting Study were preserved in
10% neutral buffered formalin.
3.8 Data Evaluation
Individual animal data presented in this report may be rounded. Therefore, use of the reported
individual values to perform any subsequent calculations may produce minor discrepancies
between the calculated values and those presented in this report.
The data collection system used for collecting the in-life and post-life data divides the study into
phases. The start of each phase begins with Day 1/Week 1. The phases presented in this report
are:
Randomization: Begins the day the animals are assigned their permanent
identification numbers. Phase days/weeks are labeled Ra1,
Ra2, Ra3, etc. Animals are not being dosed during this
phase.
Pretest: Begins for Sighting Study Animal Nos. 1001M and 1502F
due to a delay in start of the exposure. Phase days are
labeled P1, P2, P3, etc. Animals are not being dosed
during this phase.
Treatment: Begins for each Main Study animal when it receives its first
administration of test item as specified in the study plan.
Treatment 2: Begins for each Sighting Study animal when it receives its
first administration of test item as specified in the study
plan.
The following abbreviations are used in this report:
hr(s) hour(s)
M Male(s)
F Female(s)
mg/L milligrams of test item per liter of air
LC50 lethal test item concentration which would result in mortality for
50 percent of animals
FCP fixed concentration procedure

Serial Observations
Clinical Observations
Clinical observations are presented for each animal that showed signs, providing detail of the
type of sign, day or week of occurrence and information on the duration of the sign applicable.
AM Morning viability observation
PM Afternoon viability observation
PRE Prior to exposure observation
IPE Upon return to home cage observation
1Hr PE 1 hour post exposure observation
2Hr PE 2 hour post exposure observation
3Hr PE 3 hour post exposure observation
4Hr PE 4 hour post exposure observation
DEO Daily (non-exposure day) dose toxicity observation

Statistics:

No statistical analysis was required for this study.

Preliminary study:

Sighting Study: Both animals exposed to the test item at 0.05 mg/L survived until the end of
their observations period. Both animals exposed to the test item at 0.50 mg/L were euthanizedfor welfare reasons on Day 2 with clinical observations that included irregular labored breathing,
decreased activity and hunched posture. Macroscopic findings in both animals were similar and
were limited to the respiratory tract. The findings included thin red fluid in the larynx, nasal
pharynx, trachea and thoracic cavity and red discoloration of the lungs. The major factor
contributing to death was considered to be respiratory changes, based on the macroscopic
findings.

Key result

Sex:

male

Dose descriptor:

LC50

Effect level:

> 0.05 - < 0.5 mg/L air

Based on:

test mat.

Exp. duration:

4 h

Mortality:

Main Study: All animals exposed to the test item at 0.05 mg/L survived until the end of their
observations period.

Clinical signs:

irregular respiration

Body weight:

Sighting Study: There were no adverse responses following the exposure at the 0.05 mg/L
target. However, substantial decreases in bodyweights were seen on the day following the
exposure at the 0.50 mg/L target.
Main Study: There were no adverse responses following the exposure at the 0.05 mg/L target.

Gross pathology:

Sighting Study: See discussion in Section 6.2. above.
Main Study: Macroscopic changes were not present.

Other findings:

Sighting Study: There were no adverse responses following the exposure at the 0.05 mg/L
target. However, within 4 hours following the exposure at the 0.50 mg/L target, observations
that included decreased activity, hunched posture, irregular breathing and partially closed eyes
were noted. Since these observations did not abate on the day after exposure, both animals were
euthanized.
Main Study: There were no adverse responses following the exposure at the 0.05 mg/L target.

Conclusions:

The LC50 would be estimated to be >0.05 mg/L but <0.50 mg/L.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LC50

Value:

> 0.05 - < 0.5 mg/L air

Physical form:

inhalation: aerosol