7-azatridecane-1,13-diamine

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 1997-05-12 to 1997-05-16

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

Only 48 hours exposure time and no proper documentation of pH in test and control samples. The alkaline properties of the test item were not taken into account and toxicity due to alkaline pH is likely.

Qualifier:

according to guideline

Guideline:

other: "Deutsches Einheitsverfahren" DIN 38 412, Part 15, "Bestimmung der Wirkung von Wasserinhaltsstoffen auf Fische", Fischtest L15

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

GLP compliance:

yes (incl. QA statement)

Remarks:

dated 1991-10-14, according to German Chemikaliengesetz

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

The following nominal test concentrations were prepared:
range-finding test [mg/L] 1; 10; 100; 1000
main test [mg/L]: 18; 32; 58; 100
Two hours before test start 15 L of each concentration were prepared and distributed in the test containers. These solutions were stored under test conditions until the test was started. After this precoating time, the solutions were replaced by freshly prepared solutions and 10 animals were introduced into the test containers (1 test container for each concentration).
The following negative controls were prepared:
One control without additives in dilution water (1 test container with 10 fish).

Test organisms (species):

Leuciscus idus melanotus

Details on test organisms:

Species: Orfe Leuciscus idus melanotus
Source: Fischfarm Otto Schierhölter
49219 Glandorf
Length: 6.0 +/- 1.0 cm at start of test
Quality of the Fish: In the last two weeks before start of the test, no more than 5 % of animals died.
Holding:
Holding Tanks: Two round aquaria with a capacity of 400 L and an aeration system, Art. No. 01 09 00 06, supplied by: Domine GmbH Fischereitechnik
Temperature: 20 +/- 2 deg C, in a climatised room, continuous registration
Aeration: Ambient air was pumped by means of an aquarium aerator through silicon rubber tubes and "flow-out stones" into the aquaria. The air flow was adjusted in order to maintain an oxygen concentration of > 80 % of the saturation value.
Light Supply: 16 h light (200 lux - 400 lux), 8 h darkness
Holding and Dilution Water: Synthetic Water
Feeding: Trouvit Brutfutter, supplier: Milkivit-Werke, A. Trouw GmbH, 86666 Burgheim

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Post exposure observation period:

No

Hardness:

2.2 mole CaCO3/L

Test temperature:

20.1 - 20.2 deg C

pH:

8.4 - no details if measured in the control, one of the treatment groups or given as an average value

Dissolved oxygen:

92% of saturation value. No details if measured in the control, one of the treatment groups or given as an average value

Nominal and measured concentrations:

No analytical verification of test item concentration was performed, however the BHMT-fraction of the submission substance has been shown to be completely stable under similar testing conditions in the invertebrates toxicity test (IUCLID section 6.1.3, KS_Rhodia_Harlan_D54670_2012_Short-term toxicity to aquatic invertebrates).
The following nominal concentrations were tested:
range-finding test [mg/L] 1; 10; 100; 1000
main test [mg/L]: 18; 32; 58; 100

Details on test conditions:

Dilution and Holding Water
Synthetic water composed as follows:
Stock solutions:
Stock solution 1: 73.5 g CaCI2 x 2 H2O per liter solution (0.5 mole Ca 2+ /L)
Stock solution 2: 123.3 g MgSO4 x 7 H2O per liter solution (0.5 mole Mg2+ /L)
Stock solution 3: 8.40 g NaHCO3 per liter solution (0.1 mole NaHCO 3 /L)
These substances are dissolved in deionised water:
44 mL of the stock solution 1, 10 mL of the stock solution 2 and 64 mL of the stock solution 3 are combined and diluted with deionised water to 10 L.
The dilution water is aerated until a constant pH-value (8.0 +/- 0.5) is reached.

Test Containers: 18 L glass aquaria, test volume: 15 L

Test criteria:
The main test criterion was the mortality in each test solution. Those animals showing no reaction within a few seconds after touching the caudal peduncle were considered to be dead. Dead animals were removed at the observation dates (24 and 48 h after test start). At these dates, the test containers were also exchanged and cleaned from detritus. Other effects compared to the control were also noted according to a standardised form
sheet.

Test Procedure: Static test procedure, 48 h
Aeration: Ambient air was pumped by means of an aquarium aerator through silicon rubber tubes and "flow-out stones" into the aquaria. The air
flow was adjusted in order to maintain an oxygen concentration of > 80 % of the saturation.
Feeding: None
Light: 16 h light (200 Iux - 400 lux), 8 h darkness, Water Hardness: 2.2 mole CaCO3/L

Evaluation of the Results
The percentage of dead fish was calculated for each test concentration and the data transformed into "Probits". As there are no direct Probit values for 0 % and 100 %, their values for 0 % and 100 % effects were set at the values for 5 % and 95 % effects, respectively.
This is justified as the measured 0 % effect in the system used could be any value between 0 % and 10 % (one out of ten fish dead) and the measured 100 % effect could be any value between 90 % and 100 % because only 10 (not 100) animals are used. The mean values in each of these ranges are therefore chosen.

These data are plotted against the log of the nominal concentration of the test substance; the plot is fitted by means of a linear regression analysis using the following function:
Probit (Immobilisation) = m x log c + b, where
m = slope
b = section of ordinate
c = concentration of the test substance in the test container.

Reference substance (positive control):

no

Duration:

48 h

Dose descriptor:

other: NOEC and LC0

Effect conc.:

58 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: mortality and sublethal effects

Key result

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

76 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: Calculated as the geometric mean of the concentrations with 0% and 100% mortality

Duration:

48 h

Dose descriptor:

LC100

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

NOEC is the highest test concentration at which no - neither lethal nor sublethal - effect was observed compared to the control.
0 % mortality is the highest tested concentration at which the mortality of the fish during the test period was not significantly higher than the mortality in the control.
LC50 is the concentration at which 50 % of the fish were dead. This value was obtained as the geometric mean of 0 % mortality and 100 % mortality.
100 % mortality is the lowest tested concentration at which all fish were dead.

Results of the linear regression analysis
No linear regression analysis was necessary since the LC-50 could be obtained by calculating the geometric mean of 0 % mortality and 100 % mortality.

Other Effects
Besides mortality, no effects were observed.

Results with reference substance (positive control):

No reference substance

Sublethal observations / clinical signs:

Results of the Acute Toxicity Test: Cumulative number and percentage of dead fish after 24 h and 48 h

	Mortality [cumulative number of dead animals], Corresponding Percentage of Dead Animals and Probits
	Nominal concentration of the test substance [mg/L]
	0.00	18	32	58	100
Test Vessel No.:	1	2	3	4	5
Time [h]
24	0	0	0	0	10
[%]	0	0	0	0	100
Probits	3.36	3.36	3.36	3.36	6.64
48	0	0	0	0	10
[%]	0	0	0	0	100
Probits	3.36	3.36	3.36	3.36	6.64

Validity criteria fulfilled:

yes

Conclusions:

In an acute toxicity test on fish (Leucisus idus, 48h) the following effect concentrations were determined:
NOEC / LC0 (48h): 58 mg/L
LC50 (48 h): 76 mg/L
LC100 (48 h): 100 mg/L
The alkaline properties of the test item were not taken into account and toxicity due to alkaline pH is likely.

Executive summary:

The effects of the test substance "BHMT-HP - Polyamine" (BHT) on Leuciscus idus melanotus within 48 h were examined and compared to a negative control (reliability category 2, compliant to GLP). Ten animals were exposed to various concentrations of the test substance in water without using any solubilising agent.

This test was performed using a static test procedure. No analytical verification of test item concentrations were performed, however BHT has been shown to be completely stable under similar testing conditions in the invertebrates toxicity test (IUCLID section 6.1.3, SOURCE KS_Rhodia_Harlan_D54670_2012_Short-term toxicity to aquatic invertebrates).

After evaluation of the results with a "Probit"-method, the following toxicity data were obtained:

NOEC / LC0 (48h): 58 mg/L

LC50 (48 h): 76 mg/L

LC100 (48 h): 100 mg/L

No other sublethal effects were observed.

The alkaline properties of the test item were not taken into account and toxicity due to alkaline pH is likely. Thus, inherent toxicity of the test item is probably considerably lower and observed toxicity values may be taken as worst case and most probably overestimate toxicity of the test item.

Description of key information

The effects of BHT on Leuciscus idus melanotus within 48 h were examined and compared to a negative control (reliability category 2, compliant to GLP). Ten animals were exposed to various concentrations of the test substance in water without using any solubilising agent.

This test was performed using a static test procedure. No analytical verification of test item concentrations were performed, however the BHT-fraction of the substance has been shown to be completely stable under similar testing conditions in the invertebrates toxicity test (IUCLID section 6.1.3, SOURCE KS_Rhodia_Harlan_D54670_2012_Short-term toxicity to aquatic invertebrates).

After evaluation of the results with a "Probit"-method, the following toxicity data were obtained:

NOEC / LC0 (48h): 58 mg/L

LC50 (48 h): 76 mg/L

No other sublethal effects were observed.

The alkaline properties of the test item were not taken into account and toxicity due to alkaline pH is likely. Thus, inherent toxicity of the test item is probably considerably lower and observed toxicity values may be taken as worst case and most probably overestimate toxicity of the test item.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

76 mg/L