Calcium hydrogen phosphonate

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

A gene mutation in bacteria (Ames test according to OECD471) was performed with calcium hydrogen phosphonate. Results were negative with and without metabolic activation.

No other mutagenic data are available for calcium hydrogen phosphonate. A read-across with phosphonic acid is pertinent.

Gene mutation in bacteria (Ames test according to OECD471): negative with and without metabolic activation

Cytogenicity in mammalian cells (In vitro Mammalian Cell Micronucleus Test according to OECD487): negative with and without metabolic activation

Gene mutation in mammalian cells (Mouse Lymphoma Assay according to OECD476): negative with and without metabolic activation

Link to relevant study records

Reference

Reference 1

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

07 January 2016 - 11 February 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Qualifier:

according to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

Specific details on test material used for the study:

Name: CALCIUM HYDROGEN PHOSPHONATESynonyms: Calcium phosphite; Calcium hydrogenophosphonate; PHOSPHITE DE CALCIUMLot/batch No.: PCAF151028 RsAnalytical purity: 99%Expiry date:28 October 2018Storage condition: at room temperature.

Target gene:

histidine operon

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Metabolic activation system:

rat liver S9 mix

Test concentrations with justification for top dose:

The selected dose-levels were the following:- 312.5, 625, 1250, 2500 and 5000 µg/plate for the five strains in both mutagenicity experiments with and without S9 mix (except for the TA 1537 strain in the second experiment without S9 mix),- 625, 1250, 2500, 3750 and 5000 µg/plate for the TA 1537 strain in the second experiment without S9 mix.

Vehicle / solvent:

- Vehicle used: water for injections. - Justification for choice of vehicle: using a test item concentration of 50 mg/mL in the vehicle and a treatment volume of 100 µL/plate, the highest recommended dose-level of 5000 µg/plate was achievable.

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

sodium azide

benzo(a)pyrene

mitomycin C

other: 2-Anthramine

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar.DURATION- Preincubation period: 60 minutes- Incubation time: 48 to 72 hours.DETERMINATION OF TOXICITY- Method: decrease in number of revertant colonies and/or a thinning of the bacterial lawn.

Evaluation criteria:

In all cases, biological relevance (such as reproducibility and reference to historical data) was taken into consideration when evaluating the results.The test item is considered to have shown mutagenic activity in this study if:- a reproducible 2-fold increase (for the TA 98, TA 100 and TA 102 strains) or 3-fold increase (for the TA 1535 and TA 1537 strains) in the mean number of revertants compared with the vehicle controls is observed, in any strain, at any dose-level,- and/or a reproducible dose-response relationship is evidenced.The test item is considered to have shown no mutagenic activity in this study if:- neither an increase in the mean number of revertants, reaching 2-fold (for the TA 98, TA 100 and TA 102 strains) or 3-fold (for the TA 1535 and TA 1537 strains) the vehicle controls value, is observed at any of the tested dose-levels, - nor any evidence of a dose-response relationship is noted.

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Key result

Species / strain:

S. typhimurium TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

No precipitate was observed in the Petri plates when scoring the revertants at any of the tested dose levels.No noteworthy toxicity was noted at any of the tested dose-levels, towards the five strains used, either with or without S9 mix.

A slight increase in the number of revertant colonies was noted at 5000 µg/plate in the TA 1537 strain in the first experiment without S9 mix. This increase did not reach the positive threshold of 3-fold the vehicle control value (1.6-fold). However, there was an overall dose-response relationship and the mean number of revertants was above the historical data range of the corresponding vehicle control (19.3versus [2-14] for historical data). Since no noteworthy increase in the number of revertants was observed in the second experiment (under the same experimental conditions) despite the use of a narrower range of dose-levels, the slight increase observed in the first experiment was considered not to be biologically relevant.

Conclusions:

Under the experimental conditions of this study, the test item CALCIUM HYDROGEN PHOSPHONATE did not show any mutagenic activity in the bacterial reverse mutation test with Salmonella typhimurium strains, either in the presence or absence of a rat liver metabolizing system.

Executive summary:

The objective of this study was to evaluate the potential of the test item CALCIUM HYDROGEN PHOSPHONATE to induce reverse mutations in Salmonella typhimurium.

 

Methods

A preliminary toxicity test was performed to define the dose-levels of CALCIUM HYDROGEN PHOSPHONATE, suspended inwater for injections, to be used for the mutagenicity experiments. The test item was then tested in two independent experiments, both with and without a metabolic activation system, the S9 mix, prepared from a liver post-mitochondrial fraction (S9 fraction) of rats induced with Aroclor 1254.

 

Treatments were performed according to the direct plate incorporation method except for the second experiment with S9 mix, which was performed according to the pre-incubation method (60 minutes, 37°C).

 

Five strains of bacteria Salmonella typhimurium were used: TA 1535, TA 1537, TA 98, TA 100 and TA 102. Each strain was exposed to five dose-levels of the test item (three plates/dose-level). After 48 to 72 hours of incubation at, the revertant colonies were scored.

The evaluation of the toxicity was performed on the basis of the observation of the decrease in the number of revertant colonies and/or a thinning of the bacterial lawn.

 

Results

Since the test item was found to be freely soluble in the final treatment medium and non-toxic in the preliminary test, the highest dose-level selected for the main experiments was 5000 µg/plate, according to the criteria specified in the international guidelines.

 

The mean number of revertants for the vehicle and positive controls met the acceptance criteria. Also, there were five analysable dose-levels for each strain and test condition. The study was therefore considered to be valid.

 

The selected dose-levels ranged from 312.5 to 5000 µg/plate.

 

No precipitate was observed in the Petri plates when scoring the revertants at any of the tested dose-levels.

 

No noteworthy toxicity was noted at any of the tested dose-levels, towards the five strains used, either with or without S9 mix.

 

The test item did not induce any increase in the number of revertants, which could be considered as biologically relevant, in any strains or test conditions. These results met the criteria of a negative response.

 

Conclusion

Under the experimental conditions of this study, the test item CALCIUM HYDROGEN PHOSPHONATE did not show any mutagenic activity in the bacterial reverse mutation test with Salmonella typhimurium strains, either in the presence or absence of a rat liver metabolizing system.

Reference 2

Endpoint:

in vitro gene mutation study in mammalian cells

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

04-Sep-2012 to 06-Nov-2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study has been performed according to OECD and/or EC guidelines and according to GLP principles.

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.17 (Mutagenicity - In Vitro Mammalian Cell Gene Mutation Test)

Deviations:

no

Principles of method if other than guideline:

The recommendations of the “International Workshop on Genotoxicity Tests Workgroup” (the IWGT), published in the literature (Clive et al., 1995, Moore et al., 1999, 2000, 2002, 2003, 2006 and 2007).

GLP compliance:

yes (incl. QA statement)

Type of assay:

mammalian cell gene mutation assay

Target gene:

Thymidine kinase (TK) locus in L5178Y mouse lymphoma cells

Species / strain / cell type:

mouse lymphoma L5178Y cells

Details on mammalian cell type (if applicable):

- Type and identity of media:
- RPMI 1640 Hepes buffered medium (Dutch modification) containing penicillin/streptomycin (50 U/ml and 50 µg/ml, respectively), 1 mM sodium pyruvate and 2 mM L-glutamin supplemented with 10% (v/v) heat-inactivated horse serum (=R10 medium).
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: yes
- Periodically checked for karyotype stability: no
- Periodically "cleansed" against high spontaneous background: yes

Metabolic activation:

with and without

Metabolic activation system:

Rat liver S9-mix induced by a combination of phenobarbital and ß-naphthoflavone

Test concentrations with justification for top dose:

Dose range finding test:
Without and with S9-mix, 3 hours treatment: 10, 33, 100, 333, 820 µg/mL
Without S9-mix, 24 hours treatment: 10, 33, 100, 333, 820 µg/ml
Experiment 1:
Without S9-mix, 3 hours treatment: 0.3, 1, 3, 10, 33, 100, 333 and 820 µg/mL
With S9-mix, 3 hours treatment: 100, 300, 400, 500, 600, 700, 750 and 800 µg/mL
Experiment 2
Without S9-mix, 24 hours treatment: 33, 100, 400, 500, 600, 700, 750 and 820 µg/mL
With S9-mix, 3 hours treatment: 33, 100, 300, 500, 600, 700, 750 and 820 µg/mL

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: Exposure medium (RPMI 1640 Hepes buffered medium (Dutch modification) )
- Justification for choice of solvent/vehicle:

Test compound was soluble in exposure medium and exposure medium has been accepted and approved by authorities and international guidelines

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

methylmethanesulfonate

Remarks:

without S9 Migrated to IUCLID6: 15 µg/mL for the 3 hours treatment period and 5 µg/mL for the 24 hours treatment period

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

other: cyclophosphamide 10 µg/mL

Remarks:

with S9

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium

DURATION
- Exposure duration:
Short-term treatment
With and without S9-mix: 3 hours
Prolonged treatment period
Without S9-mix: 24 hours
- Expression time (cells in growth medium): 2 days
- Selection time (if incubation with a selection agent): 11 to 12 days

SELECTION AGENT (mutation assays): 5 µg/mL trifluorothymidine (TFT)

NUMBER OF REPLICATIONS:
- Solvent controls: Duplicate cultures
- Treatment groups and positive control: Single cultures

NUMBER OF CELLS EVALUATED: 9.6 x 10E5 cells plated/concentration

DETERMINATION OF CYTOTOXICITY
- Method: relative suspension growth (dose range finding test) and relative total growth (mutation experiments)

Evaluation criteria:

ACCEPTABILITY OF THE ASSAY
A mutation assay was considered acceptable if it met the following criteria:
a) The absolute cloning efficiency of the solvent controls (CEday2) is between 65 and 120%. An acceptable number of surviving cells (10^6) could be analysed for expression of the TK mutation.
b) The spontaneous mutation frequency in the solvent control is = 50 per 10^6 survivors and = 170 per 10^6 survivors.
c) The growth rate (GR) over the 2-day expression period for the negative controls should be between 8 and 32 (3 hours treatment) and between 32-180 (24 hours treatment).
d) The mutation frequency of MMS should not be below 500 per 10^6 survivors, and for CP not below 700 per 10^6 survivors.

DATA EVALUATION
Any increase of the mutation frequency should be evaluated for its biological relevance including a comparison of the results with the historical control data range.

A test substance is considered positive (mutagenic) in the mutation assay if it induces a MF of more than MF(controls) + 126 in a dose-dependent manner. An observed increase should be biologically relevant and will be compared with the historical control data range.

A test substance is considered equivocal (questionable) in the mutation assay if no clear conclusion for positive or negative result can be made after an additional confirmation study.

A test substance is considered negative (not mutagenic) in the mutation assay if:
a) None of the tested concentrations reaches a mutation frequency of MF(controls) + 126.
b) The results are confirmed in an independently repeated test.

Statistics:

The global evaluation factor (GEF) has been defined by the IWTGP as the mean of the negative/solvent MF distribution plus one standard deviation. For the micro well version of the assay the GEF is 126.

Species / strain:

mouse lymphoma L5178Y cells

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS:
- Effects of pH:
Solvent control: 7.4
820 µg/ml: 6.2
333 µg/ml: 6.9
- Effects of osmolality:
Solvent control: 0.284 mOsm/kg
820 µg/ml: 0.295 mOsm/kg
333 µg/ml: 0.291 mOsm/kg
- Precipitation: No precipitation was observed up to and including the top dose of 820 µg/mL (= 0.01 M)

RANGE-FINDING/SCREENING STUDIES:
- No toxicity was observed in the absence of S9, 3 hours treatment; Toxicity was observed at the dose level of 820 µg/mL in the presence of S9, 3 hours treatment and at dose level of 820 µg/mL in the absence of S9, 24 hours treatment

COMPARISON WITH HISTORICAL CONTROL DATA:
The spontaneous mutation frequencies in the solvent-treated control cultures were between the minimum and maximum value of the historical control data range and within the acceptability criteria of this assay.

ADDITIONAL INFORMATION ON CYTOTOXICITY:
Appropriate toxicity was observed up to and including the highest tested dose level in both experiments in the absence and presence of S9-mix.

Remarks on result:

other: strain/cell type: Test system L5178Y/TK+/-3.7.2C

Remarks:

Migrated from field 'Test system'.

Conclusions:

Interpretation of results (migrated information):
negative

Phosphonic acid (Phosphorous acid) is considered not mutagenic in the absence and presence of S9-mix in the mouse lymphoma L5178Y test system

Executive summary:

The spontaneous mutation frequencies in the solvent-treated control cultures were between the minimum and maximum value of the historical control data range and within the acceptability criteria of this assay. Except the response of the solvent control cultures in the first experiment in the presence of S9-mix. Although these responses were below the lower limit of the range, clear negative results were obtained in this part of the study, the validity of the test was considered to be not affected.

 

Mutation frequencies in cultures treated with positive control chemicals were increased 8.6 and

10-fold for in the absence of S9-mix, and 14- and 17-fold for CP in the presence of S9-mix. It was therefore concluded that the test conditions, both in the absence and presence of S9-mix, were appropriate and that the metabolic activation system (S9-mix) functioned properly.

 

In the absence of S9-mix after a 3 hours treatment period, Phosphonic acid (Phosphorous acid) did not induce a significant increase in the mutation frequency. However Phosphonic acid (Phosphorous acid) induced a 3.4-fold increase in the mutation frequency at the prolonged treatment period of 24 hours. The mutation frequency (213 x 10^-6mutant colonies) was above the global evaluation factor plus mutation frequency of the negative controls (GEF + MF_(controls): 190 x 10^-6) and outside the historical control data range. This increase was only observed at the highest tested dose level of 820 µg/ml with severe toxicity, a relative total growth of 7%.

 

In the presence of S9-mix, Phosphonic acid (Phosphorous acid) did not induce a significant increase in the mutation frequency in the first experiment. However Phosphonic acid (Phosphorous acid) induced a 5.1-fold increase in the mutation frequency in the repeat experiment. The mutation frequency (323 x 10^-6mutant colonies) was above the global evaluation factor plus mutation frequency of the negative controls (GEF + MF_(controls): 190 x 10^-6) and outside the historical control data range. This increase was only observed at the highest tested dose level of 820 µg/ml with severe toxicity, a relative total growth of 4%.

  

Since the RTG is below 10% both in the absence and presence of S9-mix, these concentrations are too toxic according to the guidelines and therefore the mutagenic responses are not biologically relevant. Furthermore, the next dose level of 750 µg/ml with a RTG of 17 and 32% in the absence and presence of S9-mix, respectively, did not even show a two-fold increase in the mutation frequency.

The dose- response was not seen as the dose levels of 700 and 750 µg/ml showed no increase in the mutation frequency more than the MF(_controls) + the GEF and were within the limit of the historical control data.Therefore, Phosphonic acid (Phosphorous acid) is considered not mutagenic in the absence and presence of S9-mix in the mouse lymphoma L5178Y test system.

Reference 3

Endpoint:

in vitro cytogenicity / micronucleus study

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

experimental study

Adequacy of study:

key study

Study period:

05-Nov-2012 to 07-Feb-2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study has been performed according to OECD and/or EC guidelines and according to GLP principles.

Qualifier:

according to guideline

Guideline:

other: OECD Guideline 487 (In Vitro Mammalian Cell Micronucleus Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

in vitro mammalian cell micronucleus test

Species / strain / cell type:

lymphocytes: human peripheral blood

Details on mammalian cell type (if applicable):

Type and identity of media:
Blood samples
Blood samples were collected by venapuncture using the Venoject multiple sample blood collecting system with a suitable size sterile vessel containing sodium heparin. Immediately after blood collection lymphocyte cultures were started.

- Culture medium
Culture medium consisted of RPMI 1640 medium, supplemented with 20% (v/v) heat-inactivated (56°C; 30 min) foetal calf serum, L-glutamine (2 mM), penicillin/streptomycin (50 U/mL and 50 µg/mL respectively) and 30 U/mL heparin.

- Lymphocyte cultures
Whole blood (0.4 mL) treated with heparin was added to 5 mL or 4.8 mL culture medium (in the absence and presence of S9-mix, respectively). Per culture 0.1 ml (9 mg/mL) phytohaemagglutinin was added.

Metabolic activation:

with and without

Metabolic activation system:

Rat liver S9-mix induced by a combination of phenobarbital and ß-naphthoflavone

Test concentrations with justification for top dose:

Dose range finding test:
Without and with S9-mix, 3hr exposure; 27 hr fixation: 33, 100, 333, 500 and 820 µg/mL
Without S9-mix, 24 exposure; 24 hr fixation: 33, 100, 333, 500 and 820 µg/mL
First cytogenetic test:
Without and with S9-mix, 3hr exposure; 27 hr fixation: 100, 333 and 820 µg/mL
Second cytogenetic test:
Without S9-mix, 24 hr exposure; 24 hr fixation: 100, 700 and 820 µg/mL

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: culture medium
- Justification for choice of solvent/vehicle:
Test compound was stable in water and soluble in culture medium. Culture medium has been accepted and approved by authorities and international guidelines

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

mitomycin C

Remarks:

without S9 Migrated to IUCLID6: MMC-C 0.25 µg/mL for a 3 hours exposure period and 0.15 µg/mL for a 24 hours exposure period

Positive control substance:

other: colchicine: 0.1 µg/mL

Remarks:

without S9

Positive control substance:

cyclophosphamide

Remarks:

with S9 Migrated to IUCLID6: 15 µg/mL

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium

DURATION
- Preincubation period: 48 hr
- Exposure duration:
Short-term treatment
Without and with S9-mix: 3 hr treatment, 24 hr recovery/harvest time
Continuous treatment
Without S9-mix: 24 hr treatment/harvest time

ARREST OF CELL DIVISION: 5 µg/mL Cytochalasine B
STAIN: Giemsa

NUMBER OF REPLICATIONS: duplicates

NUMBER OF CELLS EVALUATED: 1000/culture (mono- and binucleated cells)

DETERMINATION OF CYTOTOXICITY
- The cytostasis/cytotoxicity was determined using the cytokinesis-block proliferation index (CPBI index)

Evaluation criteria:

A test substance was considered positive (clastogenic or aneugenic) in the in vitro micronucleus test if:
a) It induces a dose-related statistically significant (Chi-square test, one-sided, p < 0.05) increase in the number of mono or binucleated cells with micronuclei.
b) A statistically significant and biologically relevant increase is observed in the number of mono or binucleated cells with micronuclei in the absence of a clear dose-response relationship.

A test substance was considered negative (not clastogenic or aneugenic) in the in vitro micronucleus test if:
a) none of the tested concentrations induced a statistically significant (Chi-square test, one-sided, p < 0.05) increase in the number of mono and binucleated cells with micronuclei.
b) The number of mono and binucleated cells with micronuclei was within the laboratory historical control data range.

Statistics:

The incidence of micronucleated cells (cells with one or more micronuclei) for each exposure group was compared to that of the solvent control using Chi-square statistics:

Species / strain:

lymphocytes: human peripheral blood

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH:
Solvent control: 7.7
820 µg/ml: 6.1
333 µg/ml: 6.8
- Effects of osmolality:
Solvent control: 287 mOsm/kg
820 µg/ml: 291 mOsm/kg

- Precipitation: No precipitation was observed up to and including the top dose of 820 µg/mL (= 0.01 M)

RANGE-FINDING/SCREENING STUDIES:
- Toxicity was only observed after the prolonged treatment period.

COMPARISON WITH HISTORICAL CONTROL DATA:
- The number of cells with chromosome aberrations found in the solvent and positive control cultures was within the laboratory historical control data range.

ADDITIONAL INFORMATION ON CYTOTOXICITY:
- T oxicity was only observed after the prolonged treatment period

Conclusions:

Interpretation of results (migrated information):
negative

Phosphonic acid (Phosphorous acid) is not clastogenic or aneugenic in human lymphocytes

Executive summary:

The number of mono- and binucleated cells with micronuclei found in the solvent control cultures in the presence of S9-mix and in the absence of S9-mix at the 24 hours exposure time was within the historical control data range per 2000 mono- or binucleated cells. In the absence of S9-mix at the 3 hours exposure time the number of binucleated cells with micronuclei of the solvent control were outside the historical control data range (8 and 9 per 1000 binucleated cells). However, the results of the test substance treated cultures were clearly negative. In addition, the positive control chemicals mitomycin C showed a statistically significant increase in the number of binucleated cells with micronuclei and colchicine showed a statistically significant increase in the number of mono- and binucleated cells with micronuclei. 

 

The positive control chemicals, mitomycin C and cyclophosphamide both produced a statistically significant increase in the number of binucleated cells with micronuclei. The positive control chemical colchicine produced a statistically significant increase in the number of mononucleated cells with micronuclei. In addition colchicine also showed a statistically significant increase in the number of binucleated cells with micronuclei. It was therefore concluded that the test conditions were adequate and that the metabolic activation system (S9-mix) functioned properly.

 

Phosphonic acid (Phosphorous acid) did not induce a statistically significant or biologically relevant increase in the number of mono- and binucleated cells with micronuclei in the absence and presence of S9-mix, in either of the two independently repeated experiments.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Genetic toxicity in vivo

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

Based on the available information, phosphonic acid does not need to be classified as mutagenic in accordance with the criteria outlined and Annex I of 1272/2002/EC.