Reaction mass of Copper,di-µ-iodotris[triphenylphosphine]di- and iodobis(triphenylphosphino)copper

Endpoint summary

Description of key information

Considering results of the structural similar compound Iodotris(triphenylphosphino)copper, the dose of 1.000 mg/kg body weight/day can be expected as the NOAEL (No Observed Adverse Effect Level) after the 28 days treatment for " Reaction mass of Copper,di-µ-iodotris[triphenylphosphine]di- and iodobis(triphenylphosphino)copper", too.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

2003

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Considering the structural similarities of the substances "Reaction mass of Copper,di-µ-iodotris[triphenylphosphine]di- and iodobis(triphenylphosphino)copper" and "Iodotris(triphenylphosphino)copper" and the almost identical physical and chemical properties, especially molecular weights, partition coefficients and water solubilites, it can be expected that the substances will show quite similar behaviour with respect to repeated dose toxicity.

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

other: 96/54/EG, B.7 (1996)

GLP compliance:

yes

Limit test:

no

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 219001
- Expiration date of the lot/batch: January 01 , 2004

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: dry, room temperature, closed container

Species:

rat

Strain:

Wistar

Details on test animals or test system and environmental conditions:

Species / Strain: rat/ Wistar Crl:WI BR
Rationale: The rat is a suitable rodent species for repeated toxicity studies, is acceptable to regulatory authorities as an indicator of potential human hazards. Extensive background data are available for this species.
Sex: male, female (nulliparous, non-pregnant)
Supplier: Charles River Wiga GmbH, D-97320 Sulzfeld, Germany
Hygiene status upon supply: SPF
Age at start of acclimatisation: 35 - 42 days
Acclimatisation: 5 (males) or 7 (females) days before start of dosing
Randomisation: The animals were allocated randomly to treatment groups using random charts one day prior to the first administration.
Mean body weight at the start of dosing:
Males: 211.9 g ± 11.3 g (5.3 %) n = 30
Females: 188.2 g ± 7.1 g (3.8 %) n = 30
Identification: ear notches; cage labelling with coloured cage cards showing the study number, dose group, time of dosing and sacrifice, animals' sex and identification number

Route of administration:

oral: gavage

Vehicle:

other: Tylose MH 1000 in deionized water

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

To determine actual concentrations, two samples of the suspensions for all three dose groups each freshly prepared and after the completion of administration were analyzed. The formulations prepared for use on day 1 and on day 24 were taken, diluted 1:100 using acetonitrile. This samples were diluted furthermore to reach the calibration range using acetonitrile after an equilibration time of 10 minutes in an ultrasonic bath and filtered using a syringe filter 0.2 μm.

The amount of the test item was analyzed by HPLC. The calibration was carried out by the external standard method.

HPLC equipment consists of a pump (Shimadzu LC-6A), Rheodyne injection valve, a column oven (Shimadzu CT0-6A), an UV /VIS detector (Shimadzu SPD-6A V) and an integrator (Shimadzu C-R4A).

The analysis was carried out under following conditions:

Column: Reprosil-Pur C18; 125 x 3 mm ID, 5 μm with guard column 10 x 3 mm
Eluent: 90 % acetonitrile / 10 % water
Flow rate: 0.5 ml/min
Pressure: 45 bar
Temperature: 30 °C
Sample loop: 20 μl
Detection: UV (250 nm)
Retention time: approx. 3.8 min

Duration of treatment / exposure:

Test duration: 28 days

Frequency of treatment:

Dosing regime: 7 days/week

Dose / conc.:

0 mg/kg bw/day (nominal)

Remarks:

control group

Dose / conc.:

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

Dose / conc.:

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

Dose / conc.:

1 000 mg/kg bw/day (actual dose received)

No. of animals per sex per dose:

Male: 10 animals at 0 mg/kg bw/day
Male: 5 animals at 300 mg/kg bw/day
Male: 5 animals at 600 mg/kg bw/day
Male: 10 animals at 1000 mg/kg bw/day
Female: 10 animals at 0 mg/kg bw/day
Female: 5 animals at 300 mg/kg bw/day
Female: 5 animals at 600 mg/kg bw/day
Female: 10 animals at 1000 mg/kg bw/day

Control animals:

yes, concurrent vehicle

Details on study design:

Sixty rats (30 males and 30 females) of the Wistar Crl:WI BR strain were allocated to four groups. Groups 2 and 3 each comprised 5 males and 5 females and groups 1 and 4 each comprised 10 males and 10 females. At the end of the 28 day treatment period 5 male and 5 female of groups 1 and 4 (groups la and 4a) were maintained, undosed, for further 14 days.

Three groups received the test item as a suspension in a watery tylose solution by oral gavage at dose levels of 300, 600 and 1000 mg/kg body weight / day. A fourth group received the vehicle only, and served as control. Animals were dosed once daily, and treatment continued for 28 days.

Observations and examinations performed and frequency:

The animals were observed daily for mortality or visible clinical signs ofreaction to treatment.

Assessments of sensory reactivity (auditory, visual and proprioceptive stimuli), grip strength and motor activity were carried out prior to administration, in the last week of dosing and in the last week of the treatment-free period.

Body weight and food consumption were recorded weekly.

Haematological parameters (erythrocyte count, haemoglobin concentration, packed cell volume, platelet count, leucocyte count and differential leucocyte count), coagulation parameters (prothrombin time and fibrinogen concentration) and clinical biochemistry parameters of the serum (alanine aminotransferase-, alkaline phosphatase-, aspartate aminotransferase-activity; protein, albumin, glucose, cholesterol, urea, creatinine, sodium and potassium content) were determined after an overnight fasting prior to killing at the termination of the study.

Sacrifice and pathology:

At termination all animals were subjected to a macroscopic pathological assessment, the organweights of the adrenals, brain, epididymides, heart, kidneys, liver, ovaries, spleen, testes and thymus were recorded and these and a range of other organs and tissues were preserved for
further histopathological examinations. The histopathological examination was performed on tissues from animals of the control group and the high dose group.

Clinical signs:

no effects observed

Description (incidence and severity):

Only in one female animal a skin lesion in the neck was observed. This skin lesion was caused by scratching itself The healing process was delayed by the necessary handling during the administration.

None of the animals showed further alterations of general state of well-being and behaviour.

The examination of sensory response, grip strength or motor activity did not show any alterations prior to the administration of the test item or at the end of the study.

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Description (incidence and severity):

Neither the body weights nor the body weight gain of the animals were influenced by the administration of the test item in comparison to the vehicle control group.

Only some significant differences were calculated. These seem to be incidental, because the body weights in the animals of the dose group were higher than the body weights in the animals of the control group or a diminished body weight increase was compensated by an increased body weight increase in the following week, respectively.

All data are in the range of the historical control data in the testing facility.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

The food consumption of the animals was not influenced by administration of the test item.
Only one significant difference was calculated (females, 2nd week). This seems to be incidental, because the difference is very small.
All data are in the range of the historical control data in the testing facility.

Food efficiency:

not examined

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

not examined

Ophthalmological findings:

not examined

Haematological findings:

effects observed, non-treatment-related

Description (incidence and severity):

None of the haematological parameters investigated was affected by the administration of the test item.
Only one significant difference was calculated (leucocytes, females, middle dose group). This seems to be incidental, because the value is in the confidence range of the historical control data in the testing facility (4.9 - 14.0·10^3 /μl).
The parameters of the leucocyte differential count were all in the normal range for the strain used.
Coagulation: None of the coagulation parameters investigated was affected by the administration of the test item.

Clinical biochemistry findings:

effects observed, non-treatment-related

Description (incidence and severity):

Only one significant difference was calculated (decreased aspartate aminotransferase activity (AST), males, high dose group), but it was not dose-dependent. This significantly decreased value was decreased in comparison to the historical control data, too.
This was the reason for the determination of this parameter also in the satellite groups. A statistical significance was not more observed, but the value was decreased, too. This decreased enzyme activity remains of uncertain toxicological relevance because a liver cell damage usually is associated with an increase in enzyme levels.

None of the other clinical-biochemical parameters investigated was affected by the administration of the test item.
The activity of the alkaline phosphatase was in all cases (also in the control animals) decreased in comparison to the historical control data. This was the reason for the determination of this parameter also in the satellite groups. Here the same effect was observed.

The cholesterol level was in all cases (also in the control animals) increased in comparison to the historical control data. This was the reason for the determination of this parameter also in the satellite groups. Here the same effect was observed.

Urinalysis findings:

not examined

Behaviour (functional findings):

not specified

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, non-treatment-related

Description (incidence and severity):

The absolute and relative organ weights of the liver were dose-dependently statistically significantly increased in all treated animals.
This increase could not be observed in the animals of the satellite groups.
The same trend but not so clearly could be observed at the absolute and relative organ weights of the kidneys. Statistically significant increases were calculated for the absolute and relative organ weight of the left kidney in the male animals of the high dose group and for the relative organ weight of the left kidney in the female animals of the low dose group.
This increase could not be observed in the animals of the satellite groups. Here the absolute and relative organ weights of the kidneys were statistically significantly decreased in the male animals of the high dose group.
None of the other absolute or relative organ weights investigated was affected by the administration of the test item.

Gross pathological findings:

no effects observed

Description (incidence and severity):

All organs and body cavities of all animals were examined with respect to the location, colour, shape and size under predominant consideration of adrenals, bone marrow (from sternum), brain, epididymides, heart, kidneys, liver, lungs, lymph nodes (mesenteric and submandibular), ovaries, prostate, sciatic nerve, small and large intestine (including Peyer's patches), spinal cord (from second trachelo-vertebra [Axis]), spleen, stomach, testes, thymus, thyroid, trachea, urinary bladder and uterus.

No abnormalities were found in any animal.

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

not examined

Histopathological findings: neoplastic:

not examined

Other effects:

not specified

Details on results:

Clinical observations:
There were no treatment-related deaths and no other
treatment-related abnormalities in clinical signs, body
weight, food consumption and neurofunctional assessment
parameters.

Laboratory findings:
A significant decrease in ASAT activity was observed in the
high dose group of male animals. There were no other
treatment-related effects on hematology or other clinical
chemistry parameters.

Effects in organs:
The absolute and relative organ weigths of the liver and
were dose-dependently increased in all dose groups. The
effect was completely reversible at the end of the recovery
period. Males of the high dose group showed increased
kidneys weights at the end of the treatment period and lower
kidney weights at the end of the recovery period.

No treatment-related histomorphological abnormality was
found in the liver, kidneys or any other organ.

Dose descriptor:

NOAEL

Effect level:

> 1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: original NCD unit is mg/kg/day.

Critical effects observed:

no

Conclusions:

Not classified

Executive summary:

The daily oral administration of the test item at doses of 300, 600 and 1.000 mg/kg body weight/day to rats for a period of 28 days was tolerated without any marked effects on physical condition.

None of the animals died during the course of investigation.

Only the following effects can be deemed as caused by the test item administration:

• Not dose-dependent decreased aspartate aminotransferase activity in the male animals of the high dose group. This effect was not completely reversible in the course of the recovery period.

This decreased enzyme activity remains of uncertain toxicological relevance because a liver cell damage usually is associated with an increase in enzyme levels.

• The absolute and relative organ weights of the liver were dose-dependently statistically significantly increased in all treated animals. The same trend but not so clearly was observed at the kidneys.

The general well-being and the behaviour were not influenced by administration of the test item. The body weight, the food consumption, the haematological and coagulation parameters were not influenced by administration of the test item. No substance-dependent macroscopic pathological and histological findings were observed.

The above mentioned results show that the test item administration caused a reversible effect to the animals also at the daily dose of 300 mg/kg bw. Therefore a NOEL (No Observed Effect Level) can not be declared.

However, based on the reversible finding considering the organ weights of liver and kidneys, the missing of other effects, especially macroscopic pathological and histological findings, the dose of 1.000 mg/kg body weight/day (dose of the high dose group) can be declared as the NOAEL (No Observed Adverse Effect Level) after the 28 days treatment.

The increase of liver and kidney weights is the result of a hyperplasia which is caused by an increased metabolic activity for the metabolism and/or elimination of the test item. A damage of cells can be excluded because an increase of enzyme activities was not observed and there were no macroscopic pathological and histological findings.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

satisfying

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion

Endpoint conclusion:

no study available