Reaction product of a ((2S)-2, 6-diaminohexanoic acid hydrochloride salt, phosphonic acid and methanal) with a 1,2-disubstituted alkane and sodium hydroxide.

Bioaccumulation: aquatic / sediment

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

bioaccumulation in aquatic species: fish

Type of information:

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

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The study was well documented and meets generally accepted scientific principles, but was not conducted in compliance with GLP.

Principles of method if other than guideline:

TEST TYPE: Accumulation and Elimination of 14C-Residues  in Carp Exposed to 14-C-CP-42902. Flow-through.

GLP compliance:

no

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

STOCK AND TEST SOLUTION AND THEIR PREPARATION:  Super stock solutions were prepared by diluting known  amounts of labelled and unlabelled test substance with  distilled water. Diluter stock solutions were prepared  by further dilution with distilled water. Test solutions  were prepared using an intermittent-flow proprtional diluter. The specific activities of the super stock solutions were  determined to be: 0.14 mg/l stock: 335 ± 24 disintegrations per minute/µg 1.4 mg/l stock solution: 33.2 ± 2.5 dpm/µg

Test organisms (species):

Cyprinus carpio

Details on test organisms:

TEST ORGANISMS: Size: 19 ± 2 cm Weight: 39 ± 3 g Pretreatment: acclimated for 30 days before testing. Cumulative mortality less than 2% during holding period Feeding: Dry pelleted ration twice daily ad libitum

Test type:

flow-through

Water / sediment media type:

natural water: freshwater

Total exposure / uptake duration:

49 d

Total depuration duration:

14 d

Hardness:

35 mg/l as CaCO3

Test temperature:

18 ± 1.0 °C

pH:

7.1

Dissolved oxygen:

> 6.0 mg/l (60% saturation)

Nominal and measured concentrations:

Nominal: 0.14 mg/l and 1.4 mg/l
Mean measured: 0.14 mg/l and 1.2 mg/l

Key result

Type:

BCF

Value:

22 dimensionless

Elimination:

yes

BIOACCUMULATION AT 0.14 mg/l NOMINAL TEST CONCENTRATION:
A mean fish concentration of 3.1 mg/kg, equivalent to BCF = 22, was determined for non-edible tissues, although no meaningful 

trends in  bioaccumulation were observed at 7 sampling points over the exposure  period. Analysis of the gut, faeces and visceral 

tissues at Day 42 showed  that 72% of residues were present in faecal matter, 16% in the gut and  12% in the remaining visceral 

tissues, indicating that the substance was  ingested and eliminated with very little accumulation.  Expressed on a  whole-body basis,

the mean fish concentration was < 0.6 mg/kg, equivalent  to BCF < 4.

BIOACCUMULATION AT 1.4mg/l NOMINAL TEST CONCENTRATION:
A similar pattern to the lower test concentration was The mean fish  concentration determined for non-edible tissues was 
12.2 mg/kg, equivalent to BCF = 10.  Analysis on Day 42 showed 45% in  faecal matter, < 5% in the gut and 50% in the remaining 

visceral tissues.  Accumulation in edible tissues was not observed.

ELIMINATION:
Elimination of the test substance from non-edible tissues was rapid and  extensive. After the 14 day depuration, 96% of accumulated 

substance was  eliminated at the 0.14 mg/l nominal test concentration; 93% was  eliminated at the 1.4 mg/l nominal test concentration.

Conclusions:

BCF values of BCF <4 - 22 were determined in a reliable study conducted according to generally accepted scientific principles.

Reference 2

Endpoint:

bioaccumulation in aquatic species: fish

Type of information:

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

Adequacy of study:

key study

Study period:

2002-03-28 to 2002-04-25

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The study was conducted according to a test protocol that is comparable to the appropriate OECD test guideline, and in compliance with GLP.

Qualifier:

according to guideline

Guideline:

other: Test Method Relating to New Chemical Substances (1974, amended 1998)

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 305 (Bioconcentration: Flow-through Fish Test)

GLP compliance:

yes

Details on sampling:

- Sampling intervals/frequency for test organisms: 4, 7, 14, 21, and 28 days

- Sampling intervals/frequency for test medium samples:0, 4, 7, 14, 21, and 28 days

- Details on sampling and analysis of test organisms and test media samples (e.g. sample preparation, analytical methods):
- Four fish were taken from each concentration level and analysed two by two. Two fish are weighed, cut into pieces, and homogenised to a slurry. 15ml Acetonitrile, 5ml purified water and 0.2 ml of triethylamine were added to 1g of slurry. The solution was homogenised, centrifuged adn the top layer collected and diluted to 50ml and then further diluted 20-fold with purified water. The solution was analysed by HPLC.
- The test media at high concentration was diluted ten-fold with purified water, then analysed by HPLC. The test media at low concentration was directly analysed by HPLC.

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

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Test substance dissolved in 2000ml purified water (High concentration level feed: 40000mg/l, low concentration feed: 4000mg/l). Each of feed solutions was supplied to a mixing glass tube by a metering pump, and diluted to their respective nominal concentrations by mixing with dilutionwater delivered by another type of metering pump, then poured into each test chamber.

Test organisms (species):

Cyprinus carpio

Details on test organisms:

TEST ORGANISM
- Common name: Carp

- Source: Sankyo Suisan Co., 1-1, Ichigayatamachi, Shinjuku-ku, Tokyo, Japan, Lot #: 01-K-1112

- Age at study initiation (mean and range, SD): Less than a year after hatching

- Length at study initiation (lenght definition, mean, range and SD): 8±4 cm

- Weight at study initiation (mean and range, SD): About 5 g

- Health status: Preacclimation: When the lot of fish was received, visual observation was made and abnormal fish were removed. The remaining fish were then reared in anaquariumwithwater flowing through. During the acclimation period, fish were kept without receiving medicine in the aquarium.

- Description of housing/holding area: aquarium

- Feeding during test: 200ml/day

ACCLIMATION

- Acclimation period: ≥2 weeks

- Type and amount of food: Babygold® from Kyorin, The food was given in an amount of 2% of the fish weight.

- Health during acclimation (any mortality observed): Mortality during the 1 week before testing: (5%

Route of exposure:

aqueous

Test type:

flow-through

Water / sediment media type:

natural water: freshwater

Total exposure / uptake duration:

28 d

Test temperature:

24±2°C

pH:

6.0-8.5

Dissolved oxygen:

260% of the saturation (25 mg/L at 240C)

Details on test conditions:

TEST MEDIUM / WATER PARAMETERS

- Intervals of test medium replacement: feed solutions were renewed after about 8 days.

OTHER TEST CONDITIONS

- Light intensity: About 16 hr/day (Hf fluorescent lamp with wavelengths of 400-700 nm)

Type:

BCF

Value:

< 10 dimensionless

Calculation basis:

steady state

Remarks on result:

other: Conc.in environment / dose:High concentration level (18.8mg/L)

Type:

BCF

Value:

< 94 dimensionless

Calculation basis:

steady state

Remarks on result:

other: Conc.in environment / dose:Low concentration level (2.03mg/l)

Details on results:

- Mortality of test organisms: <10%

- Behavioural abnormalities: There was no abnormality in shape of the body or in swimming and feeding behavior during the period of the test.

Table 1: Concentration of the test substance in the test fish and bioconcentration factor (BCF) - High concentration level

Exposure period		Fish weight (g)		Peak Area (mAbs.sec)		Conc. In final solution (mg/l)	Conc. In fish body (µg/g)	Mean conc. In water (mg/l)	BCF
Day	No.	1	2	Sample	std
4	1	4.64	8.79	<1500	19910	<0.151	<166	18.8	<9
4	2	6.94	4.84	<1500	19910	<0.151	<166	18.8	<9
7	1	4.70	6.44	<1500	19834	<0.151	<167	19.0	<9
7	2	7.49	7.41	<1500	19834	<0.151	<167	19.0	<9
14	1	6.27	5.61	<1500	17767	<0.169	<186	18.9	<10
14	2	6.82	6.40	<1500	17767	<0.169	<186	18.9	<10
21	1	5.59	6.27	<1500	17767	<0.169	<186	18.7	<10
21	2	8.74	5.87	<1500	17767	<0.169	<186	18.7	<10
28	1	6.44	5.56	<1500	17767	<0.169	<186	18.8	<10
28	2	6.46	6.75	<1500	17767	<0.169	<186	18.8	<10

Table 2: Concentration of the test substance in the test fish and bioconcentration factor (BCF) - Low concentration level

Exposure period		Fish weight (g)		Peak Area (mAbs.sec)		Conc. In final solution (mg/l)	Conc. In fish body (µg/g)	Mean conc. In water (mg/l)	BCF
Day	No.	1	2	Sample	std
4	1	9.89	4.26	<1500	19910	<0.151	<166	2.06	<81
4	2	8.16	5.76	<1500	19910	<0.151	<166	2.06	<81
7	1	10.47	5.19	<1500	19834	<0.151	<167	2.01	<83
7	2	9.18	6.50	<1500	19834	<0.151	<167	2.01	<83
14	1	10.66	4.45	<1500	17767	<0.169	<186	2.00	<93
14	2	6.00	6.56	<1500	17767	<0.169	<186	2.00	<93
21	1	4.47	7.21	<1500	17767	<0.169	<186	1.99	<94
21	2	5.84	6.35	<1500	17767	<0.169	<186	1.99	<94
28	1	7.55	5.46	<1500	17767	<0.169	<186	2.03	<92
28	2	6.40	5.00	<1500	17767	<0.169	<186	2.03	<92

Both the final concentration in water and the concentration in fish body are at approximately the limits of detection of the analytical methods for those media.

Validity criteria fulfilled:

yes

Conclusions:

Bioconcentration factors of <10 (Concentration level 18.8mg/l) and <94 (concentration level 2.03 mg/l) were determined in a reliable study conducted according to an appropriate test protocol, and in compliance with GLP.

Description of key information

SPE1415 is a complex UVCB substance. As discussed in the ‘Read-Across Justification Document’ section 13, data provided for the analogue substances are representative of SPE1415 and suitable for assessment purposes. Study data for each analogue or minor constituent has been evaluated and considered together. 

Although no studies are available for the test substance SPE1415, there have been two reliable (Klimisch 2) bioaccumulation studies in fish conducted for the analogue substances ATMP and DTPMP. These data are included in this dossier. Using a conservative approach the most sensitive study result from across the analogue substances is identified and used to address the hazard endpoint in question.

A study with ATMP (Klimisch 2) was conducted according to generally accepted scientific principles. BCF values of <4 - 22 were determined during the bioaccumulation phase of the study. After 14 -day depuration period 93 - 96% of accumulated test substance was eliminated from the test fish.

A study with DTPMP (Klimisch 2) was conducted according to GLP and generally accepted scientific principles. Bioconcentration factors of <10 and <94 were determined at nominal 18.8 mg/L and 2.08 mg/L, respectively.

The most sensitive study result has been identified as a reliable study with DTPMP (Yokohama Laboratory, 2002) which reports a bioconcentration factor of <94. Consequently, this value will be taken as the aquatic bioaccumulation endpoint for SPE1415.

Key value for chemical safety assessment

BCF (aquatic species):

94 dimensionless