Iodo(triphenylphosphino)copper

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Link to relevant study records

Reference

Reference 1

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

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 "Iodo(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 genetic toxicity in-vitro.

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

other: OECD 471 (Ames-Test)

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

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 / strain / cell type:

bacteria, other: sat TA 97a, 98, 100, 102, 1535

Metabolic activation:

with and without

Metabolic activation system:

S9-Mix (rat, PB/NF induced)

Test concentrations with justification for top dose:

Concentration range in the main test (with metabolic activation): 2 ... 200 µg/plate
Concentration range in the main test (without metabolic activation): 2 ... 200 µg/plate

Vehicle / solvent:

Solvent: demineralised water

Details on test system and experimental conditions:

Specification

species: Salmonella typhimurium L T2
strains: TA 97a, TA 98, TA 100 and TA 102, TA 1535
mutations: T A97a: his06610, TA 98: hisD3052 TA 100 and TA 1535 hisG46, TA102: hisG428, TA97, TA98 and TA100 contain uvrB; TA97, TA98, TA100 and TA102 contain pKM 101 and rfa.

Origin and Culture

Salmonella typhimurium (all strains used) were obtained from Prof. Ames and stored as stock cultures at - 80°C.
Date of arrival: June. 23rd 2001 (TA 100), Jan. 29th 2001 (TA1535) and Feb. 25th, 2002 (TA 97a, 98, 102).

Species / strain:

other: sat TA 97a, 98, 100, 102, 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Remarks:

in 1st and 2nd experiment

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Remarks:

in 1st and 2nd experiment

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Conclusions:

Interpretation of results (migrated information):
negative with metabolic activation
negative without metabolic activation

Executive summary:

RESULTS

In both experiments, the test item didn't show mutagenic effects towards Salmonella typhimurium. A concentration-based relationship could consequently not be observed, either. The test item didn't show any signs of cytotoxicity. The test item is considered as "not mutagenic under the test conditions".

DISCUSSION

The test item is considered not mutagenic. No toxic effects could be determined.

The confirmation tests of the genotype didn't show any irregularities.

In the first experiment, the titre of two strains was lower than the demanded value. As the spontaneous revertants lay within the normal range, this was stated as uncritical. The positive controls didn't evoke as many mutations as given by Prof. Ames, but the revertants were definitely increased in comparison with the negative controls, as well as showing mutagenic potential of the diagnostic mutagenes. The number of spontaneous revertants lay lower than given in the literature by Prof. Ames; however, compared with the historical values of LAUS GmbH, they were

within the normal range. For these reasons, the result of the test is considered valid.

As the test item is poorly soluble in water and other solvents, no analytical method could be developed. Therefore the content of the test item in the test solution could not be determined and the treatments tested were given as nominal concentrations.

Reference 2

Endpoint:

in vitro cytogenicity / chromosome aberration study in mammalian cells

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

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

Adequacy of study:

key study

Study period:

2004

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Considering the structural similarities of the substances "Iodo(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 genetic toxicity in-vitro.

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

other: OECD 473 (Säuger zytogenetischer in vitro-Test)

GLP compliance:

yes

Type of assay:

other: in vitro mammalian cytogenicity (B10)

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 / strain / cell type:

mammalian cell line, other: Chin. Hamster, V79-Zellen

Metabolic activation:

with and without

Metabolic activation system:

S9-mix rat, PB/NF

Test concentrations with justification for top dose:

Concentration range in the main test (with metabolic activation): 1180 ... 2000 µg/ml
Concentration range in the main test (without metabolic activation): 510 ... 2000 µg/ml

Vehicle / solvent:

Medium

Untreated negative controls:

yes

Remarks:

cell culture medium

Positive controls:

yes

Positive control substance:

ethylmethanesulphonate

Remarks:

absence of metabolic activation

Positive controls:

yes

Positive control substance:

cyclophosphamide

Remarks:

presence of metabolic activation

Details on test system and experimental conditions:

Exposure period (with metabolic activation): 4 hours
Exposure period (without metabolic activation): 18 hours

Fixation time:
18 h und 26 h

"ENGLISH"

18 h and 26 h

Species / strain:

other: as specified above

Metabolic activation:

with

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

( 2000 µg/ml)

Species / strain:

other: as specified above

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

( 710 µg/ml)

Remarks on result:

other: other: main test

Remarks:

Migrated from field 'Test system'.

Conclusions:

Interpretation of results (migrated information):
negative with metabolic activation
negative without metabolic activation

Executive summary:

The test item is considered to be non-mutagenic in the Chinese Hamster V79 cytogenetic test in vitro.

Reference 3

Endpoint:

in vitro gene mutation study in mammalian cells

Type of information:

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

Adequacy of study:

key study

Study period:

01/2017 to 12/2017

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 Iodobis(triphenylphosphino)copper and Copper,di-μ-iodotris[triphenylphosphine]di-" and "Iodo(triphenylphosphino)copper" and the almost identical physical and chemical properties, especially molecular weights, partition coefficients and water solubilites, it can be expected that these substances will show quite similar behaviour with respect to genetic toxicity in-vitro.

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)

GLP compliance:

yes (incl. QA statement)

Type of assay:

in vitro mammalian cell micronucleus test

Specific details on test material used for the study:

Identity: Iodo-bis(triphenylphosphino)copper
Trade name: Brüggolen H3301
Label name: Brüggolen ® H3301
Batch no.: 16101213

Species / strain / cell type:

Chinese hamster lung fibroblasts (V79)

Metabolic activation:

with and without

Metabolic activation system:

S9

Test concentrations with justification for top dose:

Cytotoxicity test

100.0, 50.0, 25.0, 12.5, 6.25, 3.13, 1.56, 0.781 and 0.391 μg/mL


Mutation assays

without S9 activation: 50.0, 25.0, 12.5, 6.25, 3.13 and 1.56 μg/mL
with S9 activation: 80.0, 50.0, 31.3, 19.5, 12.2 and 7.63 μg/mL


Justification for dose levels

The test item was found to be soluble in DMSO at the concentration of 50.0mg/mL, after 30minutes
of sonication at 37 °C. This solvent is known to be cytotoxic in the selected assay system
at a concentration greater than 1% (volume fraction). Based on this statement, DMSO has
to be present at a constant volume of 1% (v/v) in the negative controls and in all test item
concentrations. The addition of stock solutions at 50.0 and 25.0mg/mL in EMEM minimal
medium generated an opaque suspension with plenty precipitate; the addition of stock
solution at 10.0mg/mL generated an opaque mixture with moderate precipitate, while a
clear solution with slight precipitate was obtained in the final mixture after addition of the
test item solution at 2.50mg/mL. Based on these results, the test item was assayed at the
maximum dose level of 100 μg/mL in the preliminary cytotoxicity assay.

Vehicle / solvent:

DMSO

Untreated negative controls:

yes

Details on test system and experimental conditions:

Cytotoxicity assay

A preliminary cytotoxicity test was undertaken in order to select appropriate dose levels for
the mutation assay. In this test a wide range of dose levels of the test item was used; cell
cultures were treated using the same treatment conditions as the mutation assay, and the
survival of the cells was subsequently determined.
Treatments were performed in the absence and presence of S9 metabolism, in two separate
runs; a single culture was used at each test point and positive controls were not included.
In order to evaluate baseline count, at the beginning of treatment two additional control
cultures were included in the experimental scheme. These two cultures were trypsinized and
cell counts were performed approximately at time 0. The baseline count was the average
of the total number of cells from the two flasks. At the end of treatment, cell monolayers
were washed with PBS, the cultures were trypsinised, counted, diluted and plated. After
incubation for seven days, the colonies were stained with Giemsa solution and counted.


Mutation assay

Treatment of cell cultures
AMain Assay was performed including negative and positive controls, in the absence and
presence of S9 metabolising system. The two treatment series were assayed in separate runs.
Duplicate cultureswere prepared at each test point, with the exception of the positive controls
which were prepared in a single culture. As indicated in § 4.5, for each run, two additional
control cultures were included in the experimental scheme, in order to evaluate baseline
count. Two days before the experiment, sufficient numbers of 175cm2 flasks were inoculated
with 5 million freshly trypsinised V79 cells from a common pool, in order to have at least 20
million of cells for treatment. At the treatment time, the growth medium was removed from
the flasks and replaced with treatment medium; cultures were incubated at 37°C for three
hours.

Determination of survival (Day 0)
At the end of treatment, the medium was removed and the cell monolayers were washed with
PBS. The cultures were trypsinised, counted and an aliquot from each culture was diluted and
plated to estimate the viability of the cells. Each cell suspension was re-plated (2 x 106 cells/
F175) in order to maintain the treated cell populations. Fresh complete medium was added
to the flasks which were then returned to the incubator at 37°C in a 5% CO2 atmosphere
(100% nominal relative humidity) to allow for expression of the mutant phenotype.
4.6.3 Subculturing (Day 2, Day 5 and Day 7)
On Days 2, 5 and 7, the cell populations were subcultured in order to maintain them in
exponential growth. The cultures were trypsinised, counted and the number of cells taken
forward was adjusted to give two million viable cells seeded in 225 cm2 flasks (Day 2) or in
175 cm2 flasks (Day 5 and Day 7).

Determination of mutant frequency (Day 9)
At the expression time (Day 9), each culture was trypsinised, resuspended in complete
medium and counted by microscope. After dilution, an estimated 1 x 105 cells were plated in
each of twenty 100mmtissue culture petri dishes containing medium supplemented with
6-thioguanine (at 7.5 μg/mL). These plates were subsequently stained with Giemsa solutions
and scored for the presence of mutants. After dilution, an estimated 200 cells were plated in
each of three 60mmtissue culture petri dishes. These plates were used to estimate Cloning
Efficiency (CE).

Evaluation criteria:

Criteria for outcome of assay

A test item is considered to be clearly positive if:
– At least one of the test concentrations exhibits a statistically significant increase, compared
with the concurrent solvent/vehicle control.
– The increase is concentration-related.
– Any of the results are outside the distribution of the historical negative control data
(95% confidence limits).

A test item is considered to be clearly negative if:
– None of the test concentrations exhibits a statistically significant increase, compared
with the concurrent solvent/vehicle control.
– There is no concentration-related increase.
– All results are inside the distribution of the historical negative control data (95% confidence
limits).

Historical control data are used to demonstrate biological relevance of the results obtained.

Statistics:

The individual mutation frequency values at each test point were transformed to induce
homogeneous variance and normal distribution. The appropriate transformation was estimated
using the procedure of Snee and Irr (1981), and was found to be y = (x +a)b where
a = 0 and b = 0.275.
The mutant frequency in the solvent control and treated cultures was compared using the
Dunnett’s test (one-tailed).

Species / strain:

Chinese hamster lung fibroblasts (V79)

Metabolic activation:

with and without

Genotoxicity:

not determined

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Additional information on results:

No relevant increases in mutant numbers or mutant frequency were observed following
treatment with the test item at any dose level, in the absence or presence of S9 metabolism.

Negative and positive control treatments were included in each mutation experiment in
the absence and presence of S9 metabolism. Marked increases were obtained with the
positive control treatments, indicating the correct functioning of the assay system. For both
treatment series, the selection of the top dose and the number of analysable concentrations
were adequate and consistent with the criteria indicated in the Study Protocol.

It is concluded that Iodo-bis(triphenylphosphino)copper does not induce gene mutation in
Chinese hamster V79 cells after in vitro treatment in the absence or presence of S9 metabolic
activation, under the reported experimental conditions.

Conclusions:

Iodo-bis(triphenylphosphino)copper is not mutagenic under the reported experimental conditions.

Executive summary:

It is concluded that Iodo-bis(triphenylphosphino)copper does not induce mutation in

Chinese hamster V79 cells after in vitro treatment, in the absence or presence of S9 metabolic

activation, under the reported experimental conditions.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Justification for classification or non-classification

According to negative results in OECD 476 testing on the substance itself and on negative results in OECD 471 and 473 tests on the structural similar substance Iodotris(triphenylphosphino)copper, classification into a hazard category of the hazard class "germ cell mutagenicity" according to (EU) No. 1272/2008 is not required.