Registration Dossier

Administrative data

Description of key information

An acute oral study is not available for the substance. In an 14 -d DRF study all animals died by day 9, the first on day 5, of dosing at 1000 mg/kg bw/d (5 sex/dose). 1/5 males died and 3/5 females dies at 500 mg/kg bw/d the first by day 9 and the last by day 16, 4/10. None died at 250 mg/kg bw/d. Based on this information the substances is classified in CLP Category 4.

Based on the granulomertry an acute inhalation study is not required. In case of exposure the particles will end up in the upper airways and will be swallowed.

An acute dermal study is available. No mortality, systemic effect or effects on the skin were observed. The LD50 is > 2000 mg/kg bw.

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:
The studies were published in 1985 and 1987 and included in this review in 1991.
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Qualifier:
no guideline followed
Principles of method if other than guideline:
14-day study
Groups of 5 rats of each sex were administered 0, 62, 125, 250, 500, or 1,000 mg/kg titanocene dichloride in corn oil by gavage 5 days per week for a total of 12 dose days. Animals were weighed prior to study initiation, on days 7 and 14, and at the end of the study. Observations for signs of toxicity were made twice daily throughout the studies. Animals found moribund and those surviving to the end of the study were killed, and blood was collected for hematology and clinical chemistry analyses. A complete necropsy was performed on all animals, including those dying before the end of the study.
GLP compliance:
no
Test type:
standard acute method
Limit test:
no
Specific details on test material used for the study:
Titanocene dichloride was obtained in two lots. Lot no. PB013180 from Pfaltz and Bauer, Inc. (Waterbury, CT) was used for the 14-day and 13-week studies. Strem Chemicals (Newbury Port, MA) provided lot no. 13574-S,which was used for the 2-year studies. Identity, purity, and stability analyses were conducted at the analytical chemistry laboratory, Midwest Research Institute (MRI), Kansas City, MO. The study chemical, a dark red, microcrystalline solid, was identified as titanocene dichloride by infrared and nuclear magnetic resonance spectroscopy. Lot no.PB013180 was greater than 98% pure, as determined by titration and elemental analysis. The purity of lot no. 1357443 was determined to be greater than 99% by titration, elemental analysis, and Karl Fischer water analysis. Stability studies performed by titration indicated that titanocene dichloride was stable as a bulk chemical for at least 2 weeks at temperatures to 60°C when protected from light.
Based on the stability study results, the bulk chemical was stored at 0°C is 5°C at the testing laboratory throughout the study period. The stability of the bulk chemical was monitored by elemental analysis and by titration periodically during all phases of the studies. No change in the study material was detected.
Species:
rat
Strain:
Fischer 344
Remarks:
F344/N
Sex:
male/female
Details on test animals and environmental conditions:
14-day study

Animal Source
Charles River Breeding Laboratories, Inc., Kingston, NY

Time Held Before Study
6 or 8 days (male) 5 or 13 days (female)

Age when Placed on Study
7 week

Animals per Cage
5

Method of Animal Identification
Ear punch

Feed
NIH-07 open formula meal diet; (Zeigler Bros., Inc., Gardners, PA), available ad libitum

Maximum Storage Time for Feed
120 days after milling

Feeders
Stainless steel, gang style (Scientific Cages, Inc.,Bryan, TX), changed weekly; filled as needed

Water
Automatic watering system, (Edstrom Industries, Inc., Waterford, WI), ad libitum

Cages
Polycarbonate (Lab Products, Inc, Rochelle Park, NJ), changed twice weekly

Bedding
Aspen bed (American Co., Excelsior Baltimore, MD), changed twice weekly

Cage filters
Non-woven fiber (Snow Filtration, Cincinnati, OH)

Racks
Stainless steel (Lab Products, Inc., Ruchelle Park, NJ), changed once every two weeks

Animal Room Environment:
Temperature: 21.1°C-24.4°C
Humidity: 32%-78%
Light: fluorescent, 12 hours/day
Room air changes: 12-15 changes/hour
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
14-day study: 5 days/week for 12 dose days; sacrificed on day 17
Doses:
14-day study: 0, 62, 125, 250, 500, and 1,000 mg/kg
No. of animals per sex per dose:
14-day study: 5 males and 5 females per dose
Control animals:
yes
Details on study design:
Groups of 5 rats of each sex were administered 0, 62, 125, 250, 500, or 1,000 mg/kg titanocene dichloride in corn oil by gavage 5 days per week for a total of 12 dose days. Animals were housed five per cage. Water and feed were available ad libitum.
Animals were weighed prior to study initiation, on days 7 and 14, and at the end of the study. Observations for signs of toxicity were made twice daily throughout the studies. Animals found moribund and those surviving to the end of the study were killed, and blood was collected for hematology and clinical chemistry analyses. A complete necropsy was performed on all animals, including those dying before the end of the study. Brain, heart, right kidney, liver, lung, and thymus from all animals were weighed, as well as the right testis from all males. Portions of the heart, liver, lung, and spleen (frozen in liquid nitrogen and stored at -60°C) were taken for evaluation of tissue residues for titanium. Histopathologic examinations were performed on selected tissues and animals.

Necropsy
Complete necropsy performed on all animals. Organ weights obtained for brain, heart, right kidney, liver, lung, thymus, and right testis (males).

Histopathology
Histopathologic examinations were performed on animals dying early and on selected organs in animals at terminal sacrifice. Tissues examined microscopically included kidney, liver, stomach, and testes for males given 0 to 500 mg/kg, and kidney, liver, and stomach for females given 500 mg/kg. Control females had kidney and liver examined, and females given 250 mg/kg had kidneys examined.
Histological examinations were not performed for females given 62 or 125 mg/kg or for any animals given 1,000 mg/kg. The colon was also examined for males and females given 500 mg/kg.

Clinical Pathology
Hematology: hematocrit (automated and manual), hemoglobin, erythrocyte, leukocyte count and differential.

Clinical chemistry: blood urea nitrogen; serum creatinine, sodium, potassium, chloride, carbon dioxide, calcium, phosphorus, total protein, albumin, globulin, albumin/globulin ratio, total and direct bilirubin, cholesterol, alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase, ornithine carbamoyltransferase, sorbitol dehydrogenase, and pH.

Urinalysis: specific gravity and urine pH

Supplemental Studies
Sections of heart, liver, lungs, and spleen from control, 250 (males), and 500 mg/kg groups were evaluated for titanium residues.
Key result
Sex:
male/female
Dose descriptor:
other: Doses that lead to mortality
Effect level:
>= 500 - <= 1 000 mg/kg bw
Based on:
test mat.
Mortality:
All rats in the 1,000 mg/kg dose group died by the ninth day of dosing. In addition, four of the five male and two of the five female rats receiving 500 mg/kg titanocene dichloride died before the end of the studies.
Clinical signs:
Clinical findings in all rats in the 500 and 1,000 mg/kg dose groups included ruffled fur and hunched posture. Many of these animals also had diarrhea, ataxic gait, and a red nasal discharge. All male rats given 125, 250, 500, or 1,000 mg/kg and all females given 1,000 mg/kg were lethargic during the studies.
Body weight:
There were dose-related decreases in final mean body weight in rats given 125, 250, or 500 mg/kg titanocene dichloride. Rats given 125 mg/kg gained approximately 28% less weight than controls and rats given 250 mg/kg gained 40% to 66% less weight than controls. Rats given 500 mg/kg lost weight during the studies. Body weight gains for rats given 1,000 mg/kg were not evaluated due to high mortality in these groups.
Other findings:
Treatment-related decreases in absolute organ weights occurred for heart, kidney, lung, and thymus in male rats given 125 or 250 mg/kg titanocene dichloride, for liver in male rats given 250 mg/kg, and for heart and thymus in female rats given 125, 250, or 500 mg/kg. Female rats also showed a significant chemical-related decrease in relative thymus weights, but the ratios were significant only for the 250 mg/kg group. These changes in organ weights were related to the decreased final body weights in treated groups.
There were mild chemical related decreases in albumin, globulin, calcium, and total protein concentrations in male rats given 62, 125, or 250 mg/kg titanocene dichloride. Alanine aminotransferase (ALT) levels were significantly greater than the control for males given 62, 125, or 250 mg/kg.
Significant decreases were present in total protein levels of females given 125, 250, or 500 mg/kg and in globulin levels of female rats given 250 or 500 mg/kg. There were significant chemical-related increases in ALT levels of females given 125, 250, or 500 mg/kg, albumin/globulin ratios and direct bilirubin levels of females given 250 or 500 mg/kg, and total bilirubin levels of females given 500 mg/kg. Hematocrit, hemoglobin and erythrocyte count values were decreased in female rats in the 500 mg/kg group
Tissue samples from males given 0, 250, or 500 mg/kg and from females given 0 or 500 mg/kg were analyzed for titanium residues. The highest levels of titanium were found in the spleen and liver.
Lesions observed in treated animals included multifocal hepatocellular necrosis in two of the five males given 500 mg/kg titanocene dichloride, hepatocellular hypertrophy in all males and females given 500 mg/kg, and nephrosis (acute cortical tubule necrosis) in all rats given 500 mg/kg and in two of the five females, given 250 mg/kg. There was hyperplasia of the forestomach epithelium of most male rats given 62, 125, 250, or 500 mg/kg and in three of the five females from the 500 mg/kg group. The severity of the hyperplasia in males increased with dose. Erosions and ulcers of the glandular stomach were also present in most treated males and in females from the 500 mg/kg dose group, and were associated with acute inflammation and regenerative hyperplasia.

Survival and Mean Body Weights of Rats in the 14-Day Gavage Studies of Titanocene Dichloride

 

Dose (mg/kg bw)

Survivala

Mean Body Weightsb(g)

Final Weight Relative to Controls (%)

Initial

Final

Change

Male

0

5/5

170 ± 4

220 ± 5

50 ± 2

 

62

5/5

170 ± 7

222 ± 8

52 ± 2

101

125

5/5

169 ± 4

206 ± 5

36 ± 2

93

250

5/5

170 ± 4

187 ± 6**

17 ± 4**

85

500

1/5c

170 ± 3

107 ± 4**

69 ± 2**

48

1000

0/5d

170 ± 4

-

-

-

Female

0

5/5

94 ± 4

134 ± 5

40 ± 2

 

62

5/5

93 ± 3

138 ± 3

45 ± 2

103

125

5/5

94 ± 3

123 ± 5

29 ± 3*

91

250

5/5

94 ± 2

117 ± 2*

24 ± 2**

87

500

3/5e

94 ± 2

84 ± 10**

-8 ± 8**

63

1000

0/5f

93 ± 2

-

-

-

* Significantly different (P≤0.05) from the control group by Dunn's or Shirley's test.

**P≤0.01

a Number surviving/number initially in group

b Mean ± standard error. Subsequent calculations are based on animals surviving to the end of the study.

c Day of death: 9, 9, 9, 16

d Day of death: 5, 7, 8, 8, 9; no data reported due to 100% mortality in this group

e Day of death: 13, 16

f Day of death: 4, 7, 7, 8, 9; no data reported due to 100% mortality in this group

Interpretation of results:
Category 4 based on GHS criteria
Conclusions:
In the NTP 14-day studies, mortality occurred in both sexes of rats receiving the two highest doses (500 and 1,000 mg/kg). Animals that died were observed to have had diarrhea, ataxia, acute renal tubular necrosis, and a spectrum of degenerative lesions in the stomach. Although the precise cause of death was not determined, it is likely that the rats died from the renal lesions complicated by serum electrolyte changes or an acid-base imbalance, or both, resulting from the gastric lesions.
Based on this data the substance is classified in Category 4.
Executive summary:

Toxicology and carcinogenesis studies were conducted by administering titanocene dichloride (greater than 98% pure) in corn oil by gavage to groups of F344/N rats for 14 days.

In the 14-day studies, titanocene dichloride was administered at doses of 0, 65, 125, 250, 500, or 1,000 mg/kg. All high-dose rats and four of the five male and two of the five female rats given 500 mg/kg died during the studies. A dose-related decrease in body weight gain was seen in rats given 125, 250, 500, and 1,000 mg/kg. Lesions related to chemical administration included hepatocellular necrosis, tubule necrosis in the kidney, erosions and ulcers of the glandular stomach, and hyperplasia of the forestomach epithelium.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Quality of whole database:
One K2 study available.

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
the study does not need to be conducted because exposure of humans via inhalation is not likely taking into account the vapour pressure of the substance and/or the possibility of exposure to aerosols, particles or droplets of an inhalable size
other:
Endpoint conclusion
Endpoint conclusion:
no study available

Acute toxicity: via dermal route

Link to relevant study records
Reference
Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Experimental start date 29 November 2016 Experimental completion date 20 December 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
fixed dose procedure
Limit test:
yes
Specific details on test material used for the study:
Identification: Dichlorobis(η-cyclopentadienyl)titanium (CAS# 1271-19-8)
Batch: 07088501022
Purity: 99.3%
Physical state/Appearance: red crystalline powder
Expiry Date: 03 April 2019
Storage Conditions: room temperature in the dark
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
Animal Information
Five male and five female Wistar (RccHan:WIST) strain rats were supplied by Envigo RMS (UK) Limited, Oxon, UK. On receipt the animals were randomly allocated to cages. The females were nulliparous and non pregnant. After an acclimatization period of at least 5 days the animals were selected at random and given a number unique within the study by indelible ink marking on the tail and a number written on a cage card. At the start of the study the animals weighed at least 200 g, and were 8 to 12 weeks of age. The weight variation did not exceed ±20% of the mean weight for each sex.

Animal Care and Husbandry
The animals were housed in suspended solid floor polypropylene cages furnished with woodflakes. The initial two animals were housed individually throughout the study. The further group of eight animals (four male and four female) were housed individually during the 24 Hour exposure period and in groups of four, by sex, for the remainder of the study. Free access to mains drinking water and food (2014C Teklad Global Rodent diet supplied by Envigo RMS (UK) Limited, Oxon, UK) was allowed throughout the study. The diet, drinking water and bedding were routinely analyzed and were considered not to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study.
The temperature and relative humidity were set to achieve limits of 19 to 25 C and 30 to 70% respectively. The rate of air exchange was at least fifteen changes per hour and the lighting was controlled by a time switch to give 12 hours continuous light and 12 hours darkness.
The animals were provided with environmental enrichment items which were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.
Type of coverage:
semiocclusive
Vehicle:
arachis oil
Details on dermal exposure:
On the day before treatment the back and flanks of each animal were clipped free of hair.
In the absence of data suggesting the test item was toxic, one male and one female rat were initially treated with the test item at a dose level of 2000 mg/kg.
The appropriate amount of test item, moistened with arachis oil BP, was applied as evenly as possible to an area of shorn skin (approximately 10% of the total body surface area). A piece of surgical gauze was placed over the treatment area and semi occluded with a piece of self adhesive bandage. The animals were caged individually throughout the study. Shortly after dosing the dressings were examined to ensure that they were securely in place.
After the 24 Hour contact period the bandage was carefully removed and the treated skin and surrounding hair wiped with cotton wool moistened with arachis oil BP to remove any residual test item.
As no mortalities were noted a further group of animals (four males and four females) was similarly treated with the test item at a dose level of 2000 mg/kg body weight to give a total of five males and five females
Duration of exposure:
24 hours
Doses:
2000 mg/kg
No. of animals per sex per dose:
Five males
Five females
Control animals:
no
Details on study design:
. The animals were caged individually for the 24 Hour exposure period. After the 24 Hour contact period the bandages were carefully removed and the treated skin and surrounding hair wiped with cotton wool moistened with a suitable solvent to remove any residual test item. These animals were returned to group housing for the remainder of the test period.
The animals were observed for deaths or overt signs of toxicity 30 minutes, 1, 2 and 4 hours after dosing and subsequently once daily for 14 days.
Statistics:
3.5 Data Evaluation
Data evaluations included the relationship, if any, between the exposure of the animal to the test item and the incidence and severity of all abnormalities including behavioral and clinical observations, gross lesions, body weight changes, mortality and any other toxicological effects.
Using the mortality data obtained, an estimate of the acute dermal median lethal dose (LD50) of the test item was made.
The results were also evaluated according to Regulation (EC) No. 1272/2008, relating to the Classification, Labelling and Packaging of Substances and Mixtures and the Globally Harmonized System of Classification and Labelling of Chemicals.
3.6 Major Computerized Systems
The following computerized system was used in the study:
Delta Controls – ORCAview
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
There were no deaths.
Clinical signs:
No signs of systemic toxicity were noted during the observation period.
Body weight:
Two females showed body weight loss during the first week with expected gain in body weight during the second week. One other female showed expected gain in body weight during the first week but body weight loss during the second week. The remaining animals showed expected gains in body weight over the study period.
Gross pathology:
No abnormalities were noted at necropsy.
Other findings:
Yellow colored staining, not preventing evaluation of dermal responses, was noted the test sites of all animals, up to 6 days after dosing.
There were no signs of dermal irritation.
Interpretation of results:
GHS criteria not met
Conclusions:
The acute dermal median lethal dose (LD50) of the test item in the Wistar strain rat was found to be greater than 2000 mg/kg body weight.
Executive summary:

Introduction

The study was performed to assess the acute dermal toxicity of the test item in the Wistar strain rat.

Methods

Initially, two animals (one male and one female) were given a single, 24 hour, semi‑occluded dermal application of the test item to intact skin at a dose level of 2000 mg/kg body weight. Based on the results of the initial test, a further group of eight animals (four males and four females) was similarly treated. Clinical signs and body weight development were monitored during the study. All animals were subjected to gross necropsy.

Results

Mortality. There were no deaths.

Clinical Observations. There were no signs of systemic toxicity.

Dermal Irritation. There were no signs of dermal irritation.

Body Weight. Two females showed body weight loss during the first week with expected gain in body weight during the second week. One other female showed expected gain in body weight during the first week but body weight loss during the second week. The remaining animals showed expected gains in body weight over the study period.

Necropsy. No abnormalities were noted at necropsy.

Conclusion

The acute dermal median lethal dose (LD50) of the test item in the Wistar strain rat was found to be greater than 2000 mg/kg body weight.

The test item does not meet the criteria for classification according to Regulation (EC) No. 1272/2008, relating to the Classification, Labelling and Packaging of Substances and Mixtures and theGlobally Harmonized Systemof Classification and Labelling of Chemicals.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Quality of whole database:
One K1 study available.

Additional information

Justification for classification or non-classification

Based on this information the substance is classified in CLP Category 4 for acute oral toxicity.

Furthermore, hydrochloric acid is liberated in contact with moisture (see 7.3) and this may also happen when the substance is inhaled and hydrochloric acid is released.hydrochloric acid at C >= 10% w/w is classified as STOT Single Exp. 3. H335: May cause respiratory irritation and this hazard also needs te be taken into consideration for the registered substance.