Xanthylium, 9-(2-carboxyphenyl)-3,6-bis(diethylamino)-, molybdatesilicate

Toxicity to aquatic algae and cyanobacteria

Link to relevant study record(s)

Reference

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

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

Adequacy of study:

weight of evidence

Reliability:

1 (reliable without restriction)

Justification for type of information:

REPORTING FORMAT FOR THE ANALOGUE APPROACH

PV1:2 read-across justification to PR81:5 for IUCLID

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
This read-across is based on the hypothesis that source and target substances have the same type of toxicological effects based on common underlying mechanisms (ECHA, 2015b). All substances have the same organic structural group (colorant) with different compositions of the inorganic group (metallic salt). Read-across is in this case based on the hypothesis that source and target substances have similar toxicological properties because they degrade to the similar chemical substances (ECHA, 2015c). This prediction is supported by physicochemical and toxicological data on the substances.

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
Name Target Substance Target Substance
IUPAC name Reaction product of 9-(2-carboxyphenyl)-3,6-bis(diethylamino)xanthylium Source product of ethyl 2-[3-(ethylamino)-6-ethylimino-2,7-dimethylxanthen-9-yl]
chloride with silicomolybdic acid benzoate with silicomolybdic acid
CAS No 62973-79-9 63022-06-0
EC No 263-778-2 263-793-4
Molecular weight range ≥ 3700 ≤ 8200 ≥ 3700 ≤ 8200
Molecular formula (C28H31N2O3)x.2SiO2.(MoO3)y C28H31N2O3)x.2SiO2.(MoO3)y
x = 6-10 x = 6-10
y = 7-25 y = 7-25


Structural formula Structures are given in the document attached in the section below

Both substances are UVCB’s and so purity is considered to be 100% 
3. ANALOGUE APPROACH JUSTIFICATION
The target substance CAS 62973-79-9 is an organometallic multi-constituent substance. The source substance CAS 63022-06-0 has the same organic constituent (see table above) as the target substance consisting of a basic 3,6-bis(diethylamino)xanthylium structure which has been has additional methyl groups at positions 2,7 (PR81:5, only)and a carboxyphenyl group at position 9 which, for PR81:% only, has an ethyl substitution at position 2.. The inorganic Part B is the same between source and target substances, consisting of the elements O, Mo, P and W in different compositions. These differences are expected to behave very similar. The target and source substances are thus considered suitable for the analogue approach based on structural similarity.
4. DATA MATRIX
REPORTING FORMAT FOR THE ANALOGUE APPROACH
[Please provide information for all of the points below. Indicate if further information is included as attachment to the same record, or elsewhere in the dataset (insert links in 'Cross-reference' table)]

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
[Describe why the read-across can be performed (e.g. common functional group(s), common precursor(s)/breakdown product(s) or common mechanism(s) of action]

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
[Provide here, if relevant, additional information to that included in the Test material section of the source and target records]

3. ANALOGUE APPROACH JUSTIFICATION
[Summarise here based on available experimental data how these results verify that the read-across is justified]

4. DATA MATRIX

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

other: OECD Guideline No. 221 “Lemna sp. Growth Inhibition Test (March 2006)”

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Information as provided by the Sponsor.
Identification: Lumière Pink S.M. 8135N
Batch: 021340
CAS Number: 63022-06-0
EC Number: 263-793-4
Purity: ≥90% UVCB (treat as 100%)
Physical state/Appearance: red powder
Expiry Date: 01 July 2022
Storage Conditions: room temperature in the dark

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 0.10, 0.32, 1.0, 3.2, 10 and 32 mg/L loading rate WAF
- Sampling method: Samples were taken from the control and each loading rate WAF test group from the freshly prepared bulk test preparation on Days 0, 2 and 5 and from the corresponding old or expired pooled replicates on Days 2, 5 and 7 for quantitative analysis.
- Sample storage conditions before analysis: All samples were stored frozen prior to analysis. Duplicate samples were taken at each occasion and stored frozen for further analysis if necessary.

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
Nominal amounts of test item (20 and 200 mg) were each separately added to the surface of 2 liters of culture medium to give the 10 and 100 mg/L loading rates respectively. After the addition of the test item, the culture medium was stirred by magnetic stirrer using a stirring rate such that a vortex was formed to give a dimple at the water surface. The stirring was stopped after 23 hours and the mixtures allowed to stand for 1-Hour.

Visual observations made on the WAFs indicated that a significant amount of dispersed test item was present in the water column and hence it was considered justifiable to remove the WAFs by filtering through a glass wool plug.

Test organisms (species):

other: Lemna Gibba

Details on test organisms:

TEST ORGANISM
A culture of Lemna gibba was supplied by Bayer CropScience AG, 40789 Monheim, Germany in 2007. Cultures were maintained in the laboratory by the periodic replenishment of culture medium. The culture was maintained in the laboratory at a temperature of 24 ± 1 ºC under continuous illumination (intensity approximately 7000 lux) for at least 7 days prior to the start of the test.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

7 d

Remarks on exposure duration:

not specified

Post exposure observation period:

not specified

Hardness:

not specified

Test temperature:

24 ± 1 °C

pH:

5.4 - 6.6

Dissolved oxygen:

not specified

Salinity:

not specified

Conductivity:

not specified

Nominal and measured concentrations:

Nominal: 0.10, 0.32, 1.0, 3.2, 10 and 32 mg/L
Measured: Chemical analysis of the freshly prepared test preparations on Days 0, 3 and 5 showed measured test concentrations to range from less than the limit of quantification (LOQ) of the analytical method, determined to be 0.0020 mg/L, to 0.18 mg/L. Analysis of the corresponding old or expired test preparations on Day 2, 5 and 7 showed measured test concentrations to range from less than the LOQ to 0.12 mg/L.

Details on test conditions:

TEST SYSTEM
As in the range-finding tests 500 ml glass conical flasks were used. Three flasks each containing 250 mL of solution were prepared for the control and each treatment group.
The control group was maintained under identical conditions but not exposed to the test item.
Each control and test flask was inoculated with 3 colonies of Lemna gibba (total 12 fronds). The flasks were then incubated at 24 ± 1 ºC under constant illumination (intensity approximately 7000 lux) on a black, non-reflective surface for 7 days.
On Days 2 and 5 the test solutions were renewed.

TEST CONCENTRATIONS
- Range finding study: 10 and 100 mg/L
Based on the results of the range-finding test the following test concentrations were assigned to the definitive test: 0.10, 0.32, 1.0, 3.2, 10 and 32 mg/L loading rate WAF.

Reference substance (positive control):

yes

Key result

Duration:

7 d

Dose descriptor:

other: See table 1

Effect conc.:

ca. 0.32 - ca. 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: Table 1

Details on results:

Details on results
- Exponential growth in the control (for algal test): n/a
- Observation of abnormalities (for algal test): none
- Unusual cell shape: slight gibbosity above 3.2 mg/L
- Colour differences: some colourosis above 0.32 mg/L

Results with reference substance (positive control):

- Results with reference substance valid? yes

- EC50:
8.7 mg/L Frond number for growth rate
8.9 mg/L Dry weight for growth rate
6.4 mg/L Frond number for yield
6.0 mg/L Dry weight for yield

NOEC:
1.0 mg/L Frond number for growth rate
4.0 mg/L Dry weight for growth rate
1.0 mg/L Frond number for yield
1.0 mg/L Dry weight for yield

LOEC:
2.0 mg/L Frond number for growth rate
8.0 mg/L Dry weight for growth rate
2.0 mg/L Frond number for yield
2.0 mg/L Dry weight for yield

Reported statistics and error estimates:

Frond number

Average Specific Growth Rate:There were no statistically significant differences between the control, 0.10 and 0.32 mg/L loading rate WAFs (P≥0.05), however all other loading rates were significantly different (P<0.05) and therefore the NOEL in terms of inhibition of average specific growth rates calculated from frond numbers was 0.32 mg/L loading rate WAF. Correspondingly the LOEL was 1.0 mg/L loading rate WAF.

Yield: There were no statistically significant differences between the control and 0.10 mg/L loading rate WAF (P≥0.05), however all other loading rates were significantly different (P<0.05) and therefore the NOEL in terms of inhibition of average specific growth rates calculated from yield was 0.10mg/L loading rate WAF. Correspondingly the LOEL was 0.32 mg/L loading rate WAF.


Dry weight

Average Specific Growth Rate: There were no statistically significant differences between the control and 0.10 mg/L loading rate WAF (P≥0.05), however all other loading rates were significantly different (P<0.05) and therefore the NOEL in terms of inhibition of average specific growth rate calculated from dry weight was 0.10 mg/L loading rate WAF. Correspondingly the LOEL was determined to be 0.32 mg/L loading rate WAF.

Yield: There were no statistically significant differences between the control and 0.10 mg/L loading rate WAF (P≥0.05), however all other loading rates were significantly different (P<0.05) and therefore the NOEL in terms of inhibition of yield calculated from dry weight was 0.10 mg/L loading rate WAF. Correspondingly the LOEL was determined to be 0.32 mg/L loading rate WAF.

Table 1. Effect of PR81:5 on Lemna gibba.

Response Variable	Measurement Variable	EL50(mg/L)	No Observed Effect Loading Rate (NOEL) (mg/L)	Lowest Observed Effect Loading Rate (LOEL) (mg/L)
Average Specific Growth Rate	Frond number	38	0.32	1.0
	Dry weight	>100	0.1	0.32
Yield	Frond number	4.4	0.1	0.32
	Dry weight	2.1	0.1	0.32

Validity criteria fulfilled:

yes

Conclusions:

The effect of the test item on the growth of Lemna gibba has been investigated over a 7-Day period and gave the following results:
- EL50:
38 mg/L Frond number for growth rate
>100 mg/L Dry weight for growth rate
4.4 mg/L Frond number for yield
2.1 mg/L Dry weight for yield

NOEL:
0.32 mg/L Frond number for growth rate
0.1 mg/L Dry weight for growth rate
0.1 mg/L Frond number for yield
0.1 mg/L Dry weight for yield

LOEL:
1.0 mg/L Frond number for growth rate
0.32 mg/L Dry weight for growth rate
0.32 mg/L Frond number for yield
0.32 mg/L Dry weight for yield

Executive summary:

The growth rate LOEL was estimated to be 1.0 mg/L and the NOEL was estimated to be 0.32 mg/L from the Long term toxicity study to algae and cyanobacteria (Lemna Gibba) of source study YY55MY. As explained in the justification for the type of information, the differences in molecular structure between target and source are unlikely to lead to differences in values of the LOEL and NOEL.

Key value for chemical safety assessment

EC50 for freshwater algae:

38 mg/L

EC10 or NOEC for freshwater algae:

0.32 mg/L