reaction mass of calcium bis(dihydrogenorthophosphate) and calcium hydrogenorthophosphate

Toxicity to microorganisms

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

activated sludge respiration inhibition testing

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

REPORTING FORMAT FOR THE ANALOGUE APPROACH
See read-across justification report under Section 13 ‘Assessment Reports’.

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
In accordance with REACH Annex XI, Section 1.5, of Regulation (EC) No. 1907/2006 (REACH) the standard testing regime may be adapted in cases where a grouping or read-across approach has been applied.

The similarities may be based on:
(1) a common functional group
(2) the common precursors and/or the likelihood of common breakdown products via physical or biological processes, which result in structurally similar chemicals; or
(3) a constant pattern in the changing of the potency of the properties across the category

(1) The target substance is a reaction mass of calcium hydrogenorthophosphate and calcium bis(dihydrogenorthophosphate). The source substance of calcium hydrogenorthophosphate is therefore expected to give similar results.
(2) All members of the group will ultimately dissociate into the common breakdown products of the Ca2+ cation and the PO43-anion.
(3) Calcium orthophosphates are expected to dissociate to the ionic forms in water and therefore the toxicity results can be read across on the basis that cations are abundant in natural waters and are unlikely to contribute to toxicity.


2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
See read-across justification report under Section 13 ‘Assessment Reports’.

3. ANALOGUE APPROACH JUSTIFICATION
See read-across justification report under Section 13 ‘Assessment Reports’.

4. DATA MATRIX
See read-across justification report under Section 13 ‘Assessment Reports’.

Reason / purpose for cross-reference:

read-across: supporting information

Qualifier:

according to guideline

Guideline:

OECD Guideline 209 (Activated Sludge, Respiration Inhibition Test

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The test item was directly added to the test medium and subsequently stirred for 24 hours.

Test organisms (species):

activated sludge of a predominantly domestic sewage

Details on inoculum:

Fresh samples of activated sludge were taken from a sewage treatment plant (Ruhrverband Kläranlage, Schmallenberg, Germany) mainly fed with municipal wastewater.
- Pretreatment: the sludge was washed with isotonic solution. A stock solution of activated sludge in tap water was prepared with a mixed liquor suspended solids level of 4.6 g dry mass sludge/liter. The sludge stock solution was fed with synthetic sewage feed and was
incubated aerated overnight at 20 ± 2 °C in the dark.
- Initial biomass concentration: 1.5 g/L

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

3 h

Test temperature:

20 ± 2 °C.

pH:

8.1 - 8.2

Nominal and measured concentrations:

Nominal test substance concentrations: 10, 100, 1000 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Erlenmeyer flask, 1 L
- Material, fill volume: glass, 500 mL
- Aeration: yes
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 6 + 1 abiotic control


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: deionized water, free from inhibitory concentrations of toxic substances (e.g. Cu2+ ions)


OTHER TEST CONDITIONS
- Adjustment of pH: yes, buffered to 7.5 ± 0.5 using H2SO4 (50 g/L) solution.


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Microbial respiration rate (total oxygen uptake including heterotrophic and nitrification uptake)

TEST CONCENTRATIONS
- Range finding study
- Test concentrations: 1000 mg/L
- Results used to determine the conditions for the definitive study: EC50 > 1000 mg/L

Reference substance (positive control):

yes

Remarks:

3,5-dichlorophenol

Duration:

3 h

Dose descriptor:

other: EC20

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

inhibition of total respiration

Remarks:

respiration rate

Duration:

3 h

Dose descriptor:

EC50

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

inhibition of total respiration

Remarks:

respiration rate

Details on results:

No inhibition of the respiration rate was observed

Results with reference substance (positive control):

EC50: 2 - 25 mg/L.

Inhibition (%) of the respiration rate by the test item

Test item concentration	10 mg/L	100 mg/L	1000 mg/L
Inhibition	-14.1 %	-4.4 %	0.5 %

Validity criteria fulfilled:

yes

Conclusions:

There is no significant inhibition of bacterial respiration by Calcium bis(dihydrogenorthophosphate) up to and including a concentration of 1000 mg/L. The EC50 of the test item is > 1000 mg/L.

Reference 2

Endpoint:

activated sludge respiration inhibition testing

Type of information:

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

Adequacy of study:

key study

Justification for type of information:

REPORTING FORMAT FOR THE ANALOGUE APPROACH
See read-across justification report under Section 13 ‘Assessment Reports’.

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
In accordance with REACH Annex XI, Section 1.5, of Regulation (EC) No. 1907/2006 (REACH) the standard testing regime may be adapted in cases where a grouping or read-across approach has been applied.

The similarities may be based on:
(1) a common functional group
(2) the common precursors and/or the likelihood of common breakdown products via physical or biological processes, which result in structurally similar chemicals; or
(3) a constant pattern in the changing of the potency of the properties across the category

(1) The target substance is a reaction mass of calcium hydrogenorthophosphate and calcium bis(dihydrogenorthophosphate). The source substance of calcium hydrogenorthophosphate is therefore expected to give similar results.
(2) All members of the group will ultimately dissociate into the common breakdown products of the Ca2+ cation and the PO43-anion.
(3) Calcium orthophosphates are expected to dissociate to the ionic forms in water and therefore the toxicity results can be read across on the basis that cations are abundant in natural waters and are unlikely to contribute to toxicity.


2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
See read-across justification report under Section 13 ‘Assessment Reports’.

3. ANALOGUE APPROACH JUSTIFICATION
See read-across justification report under Section 13 ‘Assessment Reports’.

4. DATA MATRIX
See read-across justification report under Section 13 ‘Assessment Reports’.

Reason / purpose for cross-reference:

read-across: supporting information

Reason / purpose for cross-reference:

read-across source

GLP compliance:

yes (incl. QA statement)

Duration:

3 h

Dose descriptor:

other: EC20

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

inhibition of total respiration

Remarks:

respiration rate

Duration:

3 h

Dose descriptor:

EC50

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

inhibition of total respiration

Remarks:

respiration rate

Details on results:

No inhibition of the respiration rate was observed

Results with reference substance (positive control):

EC50: 2 - 25 mg/L.

Inhibition (%) of the respiration rate by the test item

Test item concentration	10 mg/L	100 mg/L	1000 mg/L
Inhibition	-14.1 %	-4.4 %	0.5 %

Validity criteria fulfilled:

yes

Conclusions:

There is no significant inhibition of bacterial respiration by Calcium bis(dihydrogenorthophosphate) up to and including a concentration of 1000 mg/L. The EC50 of the test item is > 1000 mg/L.

Description of key information

The effect of calcium bis(dihydrogenorthophosphate) on the respiration of activated sludge was tested in a study following OECD guideline 209. Activated sludge was exposed to nominal substance concentrations of 10, 100 and 1000 mg/L respectively. The oxygen consumption of the activated sludge was observed for 3 hours. An effect on the respiration rate of the activated sludge was not observed. At test termination an EC10> 1000 mg/L (nominal) was determined. Based on the available data toxic effects of calcium hydrogenorthophosphate to aquatic microorganisms are not expected.

Key value for chemical safety assessment

EC50 for microorganisms:

1 000 mg/L

EC10 or NOEC for microorganisms:

1 000 mg/L