1-Hexene, 3,3,4,4,5,5,6,6-octafluoro-6-iodo-

Repeated dose toxicity: inhalation

Administrative data

Endpoint:

short-term repeated dose toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

07 November 2013 to 04 December 2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study was conducted in compliance with OECD GLP (1997) regulations. The test method was based on OECD Guideline 412 (2009).

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Report date:

2014

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 412 (Subacute Inhalation Toxicity: 28-Day Study)

Deviations:

yes

Principles of method if other than guideline:

In this rangefinding test, exposures were conducted for 7 days with a 7 day recovery necropsy.

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

MTDID 30531

IUPAC Name:

MTDID 30531

Test material form:

other: Liquid

Details on test material:

- Name of test material (as cited in study report): MTDID 30531
- Substance type: Mono-constiuent
- Physical state: Liquid
- Analytical purity: 98.6%
- Purity test date: 10 January 2012
- Lot/batch No.: ANL1200052
- Storage condition of test material: at room temperature, protected from sunlight

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories Inc. (Portage Michigan)
- Age at study initiation: 50 days
- Weight at study initiation: Male Mean: 229.5 g, Female Mean: 173.5 g
- Fasting period before study: None (overnight fasting prior to blood collection prior to necropsy)
- Housing: INdividually in stainless steel, wire-mish cages.
- Diet (e.g. ad libitum): Certified Rodent LabDiet 5002 (PMI Nutrition International, LLC) ad libitum
- Water (e.g. ad libitum): Reverse-osmosis tap water ad libitum
- Acclimation period: 13 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.4-21.8
- Humidity (%): 43.1-50.8
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 07 November 2013 To: 04 December 2013

Administration / exposure

Route of administration:

inhalation: vapour

Type of inhalation exposure:

nose only

Vehicle:

air

Remarks on MMAD:

MMAD / GSD: Not applicable, animals were exposed to vapor.

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: A 7.9 L stainless steel, conventional nose-only exposure system (CNOS) with synthetic rubber grommets in exposure ports to engage animal holding tubes. One exposure system was dedicated for each group for the duration of the study.
- Method of holding animals in test chamber: Each animal was placed in its own holding tube for nose-only exposure.
- Source and rate of air: Breathing-quality in-house compressed air.
- Method of conditioning air: Vapors were mixed with humidified compressed air
- System of generating vapor: Vapors were genereated by metering the test substance at a known flow rate to the top of a glass bead vaporization column filled with various sized glass beads. The bead column was wrapped with an electric heating tape controlled using a J-type thermocouple and digital temperature controller set to maintain a temperature of approximately 120 C. Dry compressed air supplied to the bottom of the bead column was controlled using a regulator and measured using a flowmeter. Vaporization occured as the liquid test substance coated the surface of the heating beads as air passed up through the column. The concentrated vapors were directed through stainless steel tubing to the inlet of each exposure system, where vapors were mixed with humidified compressed air to achieve desired exposure concentration.
- Temperature, humidity, pressure in air chamber: 20-24 C, 30-70% humidity, slight negative pressure.
- Treatment of exhaust air: Exhaust air was passed through charcoal and HEPA-filtration

TEST ATMOSPHERE
- Brief description of analytical method used: Gas chromatography was utilized.
- Samples taken from breathing zone: yes

VEHICLE (if applicable)
- Justification for use and choice of vehicle: Air

Details on analytical verification of doses or concentrations:

Samples were collectted from the approximate animal-breathing zone of the exposure system via 1/8-inch stainless steel tubing. The test atmosphere samples were collected every 35 minutes automatically using an external multi-position valve. Gas sample injection onto the chromatography column occurred via an internal gas-sampling valve with a sample loop. The chromatograph was displayed, the area under the sample peak was calculated and stored, and the concentration in parts per million (ppm( was calculated.

Duration of treatment / exposure:

Animals were exposed to the test article for 6 hours a day.

Frequency of treatment:

Daily, for 7 consecutive days

Doses / concentrations

Remarks:

Doses / Concentrations:
0 (control), 55, 257, or 993 ppm
Basis:
analytical conc.

No. of animals per sex per dose:

6

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: Doses were slected based on a previously conducted acute inhalation study.
- Rationale for animal assignment (if not random): Random
- Rationale for selecting satellite groups: half of each exposure group was sacrificed at the main necropsy while the other half underwent a 7 day recovery period without exposure before necropsy.
- Post-exposure recovery period in satellite groups: 7 days

Positive control:

None

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: All animals were observed twice daily (morning and afternoon) for mortality and mribundity.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Prior to exposure, at the approximate midpoint of each exposure, and at 0 to 1 hour following exposure. During the recovery period, the animals were observed once daily at approximately the same time each day.

BODY WEIGHT: Yes
- Time schedule for examinations: Within 4 days of recept, the week prior to randomization, on the day of randomization and on Study Days 0, 3, 4, 10, and 13.

FOOD CONSUMPTION:
- Food consumption: Yes, food consumption was calculated as g/animal/day. Food was weighed on days 0, 3, 6, 7, 10 and 13.

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No

WATER CONSUMPTION: No

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: Prior to assigned scheduled necropsy (Day 7 or Day 14)
- Anaesthetic used for blood collection: Yes, isoflurane
- Animals fasted: Yes, overnight
- How many animals: All animals
- Parameters checked: Total leukocyte count, erythrocyte count, hemoglobin, hematocrit, mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, platelet count, reticulocyte count percent, absolute reticulocye count, mean platelet volume, differential leukocyte count (neutrophil, lymphocyte, monocyte, eosinophil, basophil, large unstained cell) red cell distribution width, hemoglobin distribution width, platelet estimate, red cell morphology.

CLINICAL CHEMISTRY: Yes
Prior to assigned scheduled necropsy (Day 7 or Day 14)
- Anaesthetic used for blood collection: Yes, isoflurane
- Animals fasted: Yes, overnight
- How many animals: All animals
- Parameters checked: Albumin, total protein, globulin, albumin, globulin ratio, total bilirubin, urea nitrogen, creatinine, alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase, gamma glutamyltransferase, glucose, total cholesterol, calcium, chloride, phosphorus, potassium, sodium, sorbitol dehydrogenase, triglycerides.

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: No

OTHER:

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes
Tissues examined: adrenals, kidneys, liver, lungs, testes, thymus, throid

Results and discussion

Results of examinations

Clinical signs:

effects observed, treatment-related

Mortality:

mortality observed, treatment-related

Body weight and weight changes:

effects observed, treatment-related

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Food efficiency:

not examined

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

not examined

Ophthalmological findings:

not examined

Haematological findings:

effects observed, treatment-related

Clinical biochemistry findings:

effects observed, treatment-related

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

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

Histopathological findings: neoplastic:

no effects observed

Details on results:

CLINICAL SIGNS AND MORTALITY: A single animal in the 55 ppm group was found dead but the death was not considered test article related. There were no test substance-related effects on survival. Test substance-related clinical observations for the 993 ppm group males and females included vocalization upon handling and intermittent tremors beginning 1 hour post-exposure on Day 1, yellow material on body surfaces (dorsal rump, ventral, dorsal or lateral trunk and urogential area) generally throughout the exposure period, and unkempt appearance and thin body condition on Day 6. Low or single incidences of vocalization upon handling and yellow material on dorsal rump or dorsal trunk were also noted in the 55 and/or 257 ppm group females.

BODY WEIGHT AND WEIGHT GAIN: Lower body weight gains or body weight losses were noted in the 257 ppm group males and 993 ppm group males and females throughout the exposure period and in the 55 ppm group males from Day 3 to 6 when compared to the control group. At the end of the exposure period, mean body weights were 13.7% and 17.1% lower in the 257 and 993 ppm group males, respectively and 8.8% lower in the 993 ppm group females when compared to the control group. During the recovery period, lower body weight gains were noted in the 993 ppm group males from Day 6 to 10 when compared to the control group. The 257 ppm group males and 993 ppm group females recovered from the body weight effects.

FOOD CONSUMPTION: . Lower food consumption was noted for the 257 ppm group males and the 993 ppm group males and females throughout the exposure period and in the 257 ppm group females from Day 0 to 3 compared to the control group. Food consumption was similar or higher than controls in all exposure groups during the recovery period.

FOOD EFFICIENCY: Not examined

WATER CONSUMPTION: Not examined

OPHTHALMOSCOPIC EXAMINATION: Not examined

HAEMATOLOGY: Test substance-related alterations in hematology parameters consisted of lower absolute reticulocyte counts in the 257 ppm group males and 993 ppm group males and females, higher absolute neutrophil counts in the 993 ppm group males, and lower absolute eosiniphil counts in all test substance-treated male groups on study day 7. Following the recovery interval, absolute neutrophil counts remained higher in the 993 ppm group males and females.

CLINICAL CHEMISTRY: Test substance-related alterations in serum chemistry parameters included higher alanine aminotransferase, aspartate aminotransferase, and gamma glutamyltransferase values in the 993 ppm group males and females, higher sorbitol dehydrogenase values in the 257 ppm group males and 993 ppm group males and females on Day 7. Alterations in these enzymes were not noted after the recovery interval. On Day 7, cholesterol values were below the lower limit of detection for all males and females at 993 ppm and in 1 male at 257 ppm. Cholesterol values were lower than controls in the 55 and 257 ppm group males and the 55 and 257 ppm group females. On Day 14, mean cholesterol values were higher than controls in the 993 ppm group males and females. Test substance-related lower total protein and albumin values were noted in the 993 ppm group males on Day 7. On Day 14, mean total protein, albumin, and globulin values were slightly higher and the albumin to globulin ratio was slightly lower than controls in the 993 ppm group males. Mean total protein and globulin values were slightly higher than controls in the 257 ppm group males. Total T3 values were lower than controls in all test substance-treated male and female groups on Day 7. On Day 14, mean total T3 values were higher than controls in the 993 ppm group males. Total T4 values were higher than controls in all test substance-treated male and female groups on Day 7. On Day 14, T4 values were higher than controls in the 993 ppm males and females. Mean TSH values were higher in the 993 ppm group males and females on Day 7. Individual TSH values were higher than the upper limit of instrument evaluation for all males at 257 ppm and for one male at 55 ppm.

URINALYSIS: Not examined

NEUROBEHAVIOUR: Not examined

ORGAN WEIGHTS: Exposure to 257and 993 ppm test article was associated with higher liver and adrenal gland weights and lower thymus weights in males and females and lower testes weights in males. Exposure to 993 ppm was associated with higher kidney weights in males and females. The adrenal gland and kidney weight changes were reversible after a 7-day recovery (non-exposure) period. The liver, thymus and testes weight changes were not completely reversible after recovery.

GROSS PATHOLOGY:Exposure to 993 ppm of the test article was associated with pallor in the liver and yellow matting of the skin in males and females. Exposure to 257 and 993 ppm was associated with enlargement and dark red discoloration of the adrenal glands in males and females.

HISTOPATHOLOGY: NON-NEOPLASTIC: Microscopically, enlargement correlated with adrenal cortical hypertrophy and dark red areas correlated with hemorrhage. Exposure to all dose levels was associated with midzonal hepatocellular vacuolation, single cell necrosis, and increased mitoses in the liver. Exposure to 257 and 993 ppm was associated with hypertrophy, hemorrhage, necrosis and acute inflammation in the adrenal cortex; lymphoid depletion in the thymus; and germ cell degeneration in the testes. Hepatocellular vacuolation, single cell necrosis, and subacute inflammation were observed in livers from males and females at all exposure levels (except single cell necrosis at 55 ppm) after a 7-day recovery period, indicating a lack of reversibility in this time period. Adrenal cortical hemorrhage and seminiferous tubular degeneration of the testes were also observed at the recovery necropsy with exposure to 993 ppm and thymic lymphoid depletion was observed at the recovery necropsy with exposure to 257 ppm and 993 ppm.

HISTOPATHOLOGY: NEOPLASTIC (if applicable): None

Effect levels

Dose descriptor:

NOAEC

Effect level:

ca. 0.79 mg/L air (analytical)

Based on:

other: analytical concentration from breathing space

Sex:

male/female

Basis for effect level:

other: overall effects, clinical signs, body weight; haematology; clinical chemistry; organ weights; histopathology

Target system / organ toxicity

Critical effects observed:

not specified

Applicant's summary and conclusion

Conclusions:

Based on the results of the study, the No Observed Adverse Effect Concentration (NOAEC) was 55 ppm (0.79 mg/L, vapor) for the test article.

Executive summary:

The repeat-dose inhalation toxicity potential of the test article (Clear and colorless to pale pink liquid, Purity 98.6%, Lot 2) was evaluated in a 1-week screening study in Sprague-Dawley rats. The study was conducted in compliance with GLP regulations. The test method was based on OECD 412 (2009). Rats (6/sex/dose) were exposed nose-only to 0, 55, 257, or 993 ppm (0, 0.79, 3.72, 14.38 mg/L) of the test article vapor 6 hours/day for 7 consecutive days. Clinical observations (prior to exposure, at exposure midpoint, once daily thereafter) and body weights (Days 0, 3, 6, 10, 13, and 14) were noted. Hematology, serum chemistry, and serum hormone levels were analyzed for all rats (3/sex/group) at the primary (Day 7) and recovery (Day 14) necropsies. Complete necropsies were conducted on all animals and the adrenals, kidneys, liver, lungs, testes (males) thymus and thyroids were examined in control and 993 ppm group animals at necropsy. Gross lesions and target tissues were examined from all animals at primary and recovery necropsies. A single animal in the 55 ppm group was found dead but the death was not considered test article related. There were no test substance-related effects on survival. Test substance-related clinical observations for the 993 ppm group males and females included vocalization upon handling and intermittent tremors beginning 1 hour post-exposure on Day 1, yellow material on body surfaces (dorsal rump, ventral, dorsal or lateral trunk and urogential area) generally throughout the exposure period, and unkempt appearance and thin body condition on Day 6. Low or single incidences of vocalization upon handling and yellow material on dorsal rump or dorsal trunk were also noted in the 55 and/or 257 ppm group females. Lower body weight gains or body weight losses were noted in the 257 ppm group males and 993 ppm group males and females throughout the exposure period and in the 55 ppm group males from Day 3 to 6 when compared to the control group. At the end of the exposure period, mean body weights were 13.7% and 17.1% lower in the 257 and 993 ppm group males, respectively and 8.8% lower in the 993 ppm group females when compared to the control group. During the recovery period, lower body weight gains were noted in the 993 ppm group males from Day 6 to 10 when compared to the control group. The 257 ppm group males and 993 ppm group females recovered from the body weight effects. Lower food consumption was noted for the 257 ppm group males and the 993 ppm group males and females throughout the exposure period and in the 257 ppm group females from Day 0 to 3 compared to the control group. Food consumption was similar or higher than controls in all exposure groups during the recovery period. Test substance-related alterations in hematology parameters consisted of lower absolute reticulocyte counts in the 257 ppm group males and 993 ppm group males and females, higher absolute neutrophil counts in the 993 ppm group males, and lower absolute eosinophil counts in all test substance-treated male groups on study day 7. Following the recovery interval, absolute neutrophil counts remained higher in the 993 ppm group males and females. Test substance-related alterations in serum chemistry parameters included higher alanine aminotransferase, aspartate aminotransferase, and gamma glutamyltransferase values in the 993 ppm group males and females, higher sorbitol dehydrogenase values in the 257 ppm group males and 993 ppm group males and females on Day 7. Alterations in these enzymes were not noted after the recovery interval. On Day 7, cholesterol values were below the lower limit of detection for all males and females at 993 ppm and in 1 male at 257 ppm. Cholesterol values were lower than controls in the 55 and 257 ppm group males and the 55 and 257 ppm group females. On Day 14, mean cholesterol values were higher than controls in the 993 ppm group males and females. Test substance-related lower total protein and albumin values were noted in the 993 ppm group males on Day 7. On Day 14, mean total protein, albumin, and globulin values were slightly higher and the albumin to globulin ratio was slightly lower than controls in the 993 ppm group males. Mean total protein and globulin values were slightly higher than controls in the 257 ppm group males. Total T3 values were lower than controls in all test substance-treated male and female groups on Day 7. On Day 14, mean total T3 values were higher than controls in the 993 ppm group males. Total T4 values were higher than controls in all test substance-treated male and female groups on Day 7. On Day 14, T4 values were higher than controls in the 993 ppm males and females. Mean TSH values were higher in the 993 ppm group males and females on Day 7. Individual TSH values were higher than the upper limit of instrument evaluation for all males at 257 ppm and for one male at 55 ppm. Exposure to 993 ppm of the test article was associated with pallor in the liver and yellow matting of the skin in males and females. Exposure to 257 and 993 ppm was associated with enlargement and dark red discoloration of the adrenal glands in males and females. Microscopically, enlargement correlated with adrenal cortical hypertrophy and dark red areas correlated with hemorrhage. Pallor of the liver was also observed after a 7 day recovery period. Exposure to 257 and 993 ppm test article was associated with higher liver and adrenal gland weights and lower thymus weights in males and females and lower testes weights in males. Exposure to 993 ppm was associated with higher kidney weights in males and females. The adrenal gland and kidney weight changes were reversible after a 7-day recovery (non-exposure) period. The liver, thymus and testes weight changes were not completely reversible after recovery. Exposure to all dose levels was associated with midzonal hepatocellular vacuolation, single cell necrosis, and increased mitoses in the liver. Exposure to 257 and 993 ppm was associated with hypertrophy, hemorrhage, necrosis and acute inflammation in the adrenal cortex; lymphoid depletion in the thymus; and germ cell degeneration in the testes. Hepatocellular vacuolation, single cell necrosis, and subacute inflammation were observed in livers from males and females at all exposure levels (except single cell necrosis at 55 ppm) after a 7-day recovery period, indicating a lack of reversibility in this time period. Adrenal cortical hemorrhage and seminiferous tubular degeneration of the testes were also observed at the recovery necropsy with exposure to 993 ppm and thymic lymphoid depletion was observed at the recovery necropsy with exposure to 257 ppm and 993 ppm.  Based on the results of the study, the No Observed Adverse Effect Concentration (NOAEC) was 55 ppm (0.79 mg/L, vapor) for the test article.