Octasodium 2-(8-(4-chloro-6-(3-((4-chloro-6-(3,6-disulfonato-2-(1,5-disulfonatonaphthalen-2-ylazo)-1-hydroxynaphthalen-8-ylamino)-1,3,5-triazin-2-yl)aminomethyl)phenylamino)-1,3,5-triazin-2-ylamino)-3,6-disulfonato-1-hydroxynaphthalen-2-ylazo)naphthalene-1,5-disulfonate

Bioaccumulation: aquatic / sediment

Administrative data

Endpoint:

bioaccumulation in aquatic species: fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

08 March 1993 to 31 March 1993

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted to recent EU test guidance in compliance with GLP.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1993

Report date:

1993

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 305 C (Bioaccumulation: Test for the Degree of Bioconcentration in Fish)

Deviations:

no

Principles of method if other than guideline:

The study was conducted following "Bioaccumulation test of chemical substances in fish body" laid down in "the Test requirement on New chemical substances" (KNHOGTO No.5, YAKUHATSU No.615, 49-KIKYOKtf No.392, dated on 13 Jul 1974), and also "305C, Bioaccumulation : Degree of Bioaccumulation in Fish" in "OECD Guidelines for Testing of chemicals" (dated on 12 May 1981).The study was conducted in accordance with the Principles of Good Laboratory Practice (GLP)

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Octasodium 2-(8-(4-chloro-6-(3-((4-chloro-6-(3,6-disulfonato-2-(1,5-disulfonatonaphthalen-2-ylazo)-1-hydroxynaphthalen-8-ylamino)-1,3,5-triazin-2-yl)aminomethyl)phenylamino)-1,3,5-triazin-2-ylamino)-3,6-disulfonato-1-hydroxynaphthalen-2-ylazo)naphthalene-1,5-disulfonate

EC Number:

413-550-5

EC Name:

Octasodium 2-(8-(4-chloro-6-(3-((4-chloro-6-(3,6-disulfonato-2-(1,5-disulfonatonaphthalen-2-ylazo)-1-hydroxynaphthalen-8-ylamino)-1,3,5-triazin-2-yl)aminomethyl)phenylamino)-1,3,5-triazin-2-ylamino)-3,6-disulfonato-1-hydroxynaphthalen-2-ylazo)naphthalene-1,5-disulfonate

Cas Number:

142068-96-0

Molecular formula:

Hill formula: C53H28Cl2N14Na8O26S8

IUPAC Name:

octasodium 5-[(4-chloro-6-{[4-({[4-chloro-6-({7-[2-(1,5-disulfonatonaphthalen-2-yl)diazen-1-yl]-8-hydroxy-3,6-disulfonatonaphthalen-1-yl}amino)-1,3,5-triazin-2-yl]amino}methyl)phenyl]amino}-1,3,5-triazin-2-yl)amino]-3-[2-(1,5-disulfonatonaphthalen-2-yl)diazen-1-yl]-4-hydroxynaphthalene-2,7-disulfonate

Details on test material:

- Name of test material (as cited in study report): H112339
- Physical state: Dark red powder
- Analytical purity: 84.66 wt% (in total)
- Impurities (identity and concentrations): Organic minor components in total 3.64wt%, Sodium Chloride 1.2wt%, Water 10.5 wt%
- Lot/batch No.: ASG 9108127
- Stability under test conditions: confimed at preliminary experiments prior to exposure
- Storage condition of test material: cool and dark place

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
Not applicable.

Radiolabelling:

no

Sampling and analysis

Details on sampling:

Water samples for analysis were taken at 4 times in total, twice per week from test tank as one sample at each sampling time.

Fish analysis was conducted at Week 2, using 2 fish from test tank. Control fish were analysed when the exposure started and when the study completed by sampling 2 fish for each analysis.

Test solutions

Vehicle:

no

Details on preparation of test solutions, spiked fish food or sediment:

The required amount of test substance was weighed and dissolved into deionised water to prepare 677 mg/l stock solution. It was then supplied to test tank from 25 litre volume glass tank for stock solution. Based on 48 hours LC 50 data and the sensitivity at the analysis on test substance, the nominal test concentration was 1.69 mg/l. Concurrent blank vessels were used as controls

Test organisms

Test organisms (species):

Cyprinus carpio

Details on test organisms:

Species: Carp fevnrinus carpio)
Supplier: Mango Fishfarm centre (Shiga prefecture)
Fish arrived on 15 Dec 92

Breeding conditions
Duration : After removing any abnormal fish at gross examination, fish were treated in a chemical bath and then retained for 27 days at running flow.
chemical bath : Fish were treated for 24 hours in chemical bath of 50 mg/1 Teramycin (Taito Fizer Co.) and 7 g/1 Sodium Chloride at static flow

Acclimatisation
While fish were acclimatised at acclimatisation tank, any abnormal fish were removed. The rest of fish were retained for 27 days at running flow at 25±2°c. After moved into test vessels, for further 20 days, fish were bred at the same temperature at running flow.
Acclimatisation completed : on 19 Feb 93

Weight average 22.0 gr
Length average 9.7 cm
Fat content : average 3.8%

Diets
Type formulated pellet diets for fish Supplier Nihon Haigo Shiryo Co.

Feeding conditions : Diets equivalent to approximately 2% of fish weight was fed in total, twice per day, half at each time. On the day before the termination, no diets was given to fish.

Study design

Route of exposure:

aqueous

Test type:

semi-static

Water / sediment media type:

natural water: freshwater

Total exposure / uptake duration:

2 wk

Test conditions

Hardness:

120 mg/l

Test temperature:

25 ±2°C

pH:

7.4 - 7.9

Dissolved oxygen:

0.7mg/l

TOC:

DOC as follows:
at exposure started : 7.7 mg/l
at exposure completed : 5.9 mg/l

Salinity:

Not applicable.

Details on test conditions:

Test vessel; 1001 volume glass water vessel
Flow rate: stock solution of test sample was put into test vessels at 2mi/min rate and test water was supplied at 800ml/min at 1155 l/day rate.
Test temperature: 25+2°C
DOC concentration: Test solution: 7.1 - 7.7 mg/1
Control: 7.5 - 7.8 mg/1
Number of fish used: Test level: 5 (at start)
Control: 3 (at start)
Test duration: 2 weeks

Nominal and measured concentrations:

1.69 mg/l

Reference substance (positive control):

no

Details on estimation of bioconcentration:

Details in table form submitted under any other information, results section

Results and discussion

Bioaccumulation factor

Type:

BCF

Value:

< 3.3 dimensionless

Basis:

other: Test solution

Remarks on result:

other: No further data available from study submitted

Remarks:

Conc.in environment / dose:1.69 mg/l

Details on kinetic parameters:

Not measured.

Metabolites:

Not measured.

Results with reference substance (positive control):

Not applicable.

Details on results:

The level of accumulation of test substance in carp was all below 3.3 times.

There was no abnormality in test fish by gross observation.

The mean concentration of test substance in test water has been kept constantly at the nominal levels .

Reported statistics:

No data

Any other information on results incl. tables

Calculation table for analysis of test water

Sample description	A	B	C
Standard 1.69 mg/l	30302
Test water at 4^thday	29184	1.63
Standard 1.69 mg/l	29059
Test water at 7^thday	26521	1.54
Standard 1.69 mg/l	30553
Test water at 11^thday	30666	1.70
Standard 1.69 mg/l	27361
Test water at 14^thday	27716	1.71	1.64
A: Peak area (μV-sec)
A(std): Standard solution		A(t): Sample
B: Concentration of test substance in test water (mg/l)
	B=PxA(l)/A(std)
C: Average concentration of test substance in test water (mg/l)
	C=(C(1)...C(n)/n
	n: Number of test water analysis
C(1): First analysis of test water		C(n): Last analysis of test water
P: Concentration of standard solution		1.69 mg/l

 

Calculation table for recovery and blank test (Analysis of test fish)

Sample description		A	B	C	D	E	G
Standard 1.69 mg/l		39793
Recovery a		31845	1/150	20	-	4060	79.9%
Recovery b		30854	1/150	20	-	3930	77.4%
Standard 1.69 mg/l		40135
Blank a		n.d.	1/150	20	-	-
Blank b		n.d.	1/150	20	-	-
a,b: Individual sample
A: Peak area (μV-sec)
	A(std): Standard solution			A(t): Sample
B: Ratio of portion used for analysis
C: Final volume (ml)
D: Amount of blank in test fish (μg)
E: Amount of test substance recovered (μg)
	E = PxA(l)/A(std)x1/BXC-D
F: Recovery rate (%)
	F = E/Qx100
P: Concentration of test substance in standard solution (mg/l)
Q: Amount of test substance added (5.08mg)

 

Calculation table for analysis of test fish

Sample description		A	D	G	K	H	J
Standard 1.69 mg/l		40845
Test fish after 2 weeks a		n.d.	1	23.3	-	1.64	-
Test fish after 2 weeks b		n.d.	1	26.1	-	1.64	-
A: Peak area (μV-sec)
	A(std): Standard solution			A(t): Sample
B: Ratio of portion used for analysis					1/150
C: Final volume			20ml
D: Dilution factor
E: Average concentration of blank in analysis of control
F: Recovery rate			78.6%
G: Weight of test fish (g)
K: Concentration of test substance in test fish (μg/g)
	K=(PxA(t)/A(std)/BxDxC/G-E)/Fx100
H: Average concentration of test substance in test water (mg/l)
J: BCF
	J=(PxA(t)/A(std)/BxDxC/G-E)/Fx100/H
P: Concentration of test substance in standard solution						1.69 mg/l

 

Calculation table for analysis of test fish (Control)

Sample description		A		E		G		I
Standard 1.69 mg/l		42268
Before the initiation of exposure a		n.d.		-		21.7		-
Before the initiation of exposure b		n.d.		-		22.6
Standard 1.69 mg/l		40036
After the termination of exposure a		n.d.		-		25.7		-
After the termination of exposure b		n.d.		-		22.8		-
A: Peak area (μV-sec)
	A(std): Standard solution				A(t): Sample
B: Ratio of portion used for analysis					1/150
C: Final volume					20ml
E: Amount of blank analysis of control (μg)
	E=PxA(t)/A(std)x/BxC
G: Weight of test fish (g)
I: Concentration of blank in test fish (μg/g)
	I=E/G
P: Concentration of test substance in standard solution					1.69 mg/l

 

Applicant's summary and conclusion

Conclusions:

The level of accumulation of test substance in carp was all below 3.3 times.

Executive summary:

The purpose of this study was aimed to investigate the level of bioaccumulation factors of H 112339 in carp.

 

The level of, accumulation of test substance in carp was all below 3.3 times.

 

There was no abnormality in test fish by gross observation.

 

The substance is not considered to have the potential to bioaccumulate on the basis of the study conducted.