2,4,6-trinitrotoluene

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

The preliminary tests were performed on March 26 – 28 and 29 – 31. 2011. The definitive test was performed on April 13 - 15. 2011

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

yes

Remarks:

Temperature registered in the definitive test was in the range of 18.4 – 19.6 °C with variation of 1.2°C. Therefore, the variation was greater than recommended by the Guideline.

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Test organisms (species):

Daphnia magna

Test type:

static

Water media type:

freshwater

Total exposure duration:

48 h

Remarks on exposure duration:

Basic solution was shaken for 93 h before initiation of the study

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

9.49 mg/L

Nominal / measured:

nominal

Conc. based on:

dissolved

Remarks:

Conc: 0.39; 0.78; 1.56; 3.125; 6.25; 12,5; 25 mg/l.

Basis for effect:

other: immobility

Remarks on result:

other: 4.42 – 21.15

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

6.25 mg/L

Nominal / measured:

nominal

Conc. based on:

dissolved

Remarks:

Conc. 0.39; 0.78; 1.56; 3.125; 6.25; 12,5; 25 mg/l

Basis for effect:

other: immobility

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

12.5 mg/L

Nominal / measured:

nominal

Conc. based on:

dissolved

Remarks:

Conc. 0.39; 0.78; 1.56; 3.125; 6.25; 12,5; 25 mg/l

Basis for effect:

other: immobility

In the definitive testDaphnia magnawas exposed to the test item in seven test concentrations 0.39; 0.78; 1.56; 3.125; 6.25; 12.5 and 25 mg/Lfor 48 h. The results of immobilization observations performed after 24 and 48 h are presented in Table 6.In test concentrations 0.39; 0.78; 1.56 and 6.25 mg/L no immobilization of daphnids was observed. However, in test concentrations 3.125; 12.5 and 25 mg/L the observed immobilization of daphnids was 10; 75 and 100%, respectively.In the control no immobilization was observed.

The temperature recorded was in the range of 18.4 – 19.6°C (Figure 1). During the experiment the range of variation was1.2 ºC.

Nominal concentration of test item [mg/L]	Number of tested organisms	Number of immobilized								Total percentage of immobilized
		24 h				48 h
		Replicates
		A	B	C	D	A	B	C	D	24 h	48 h
Control	20	0	0	0	0	0	0	0	0	0	0
0.39	20	0	0	0	0	0	0	0	0	0	0
0.78	20	0	0	0	0	0	0	0	0	0	0
1.56	20	0	0	0	0	0	0	0	0	0	0
3.125	20	0	0	0	1	0	0	0	2	5	10
6.25	20	0	0	0	0	0	0	0	0	0	0
12.5	20	1	3	0	1	4	4	4	3	25	75
25	20	3	3	4	2	5	5	5	5	60	100

Toxicity of 2,4,6-trinitrotoluen (TNT)forDaphnia magnaafter 48 h

Nominal concentration of test item [mg/L]	Control		0.39	0.78	1.56	3.125	6.25	12.5	25
Immobilization [%]	0.0		0.0	0.0	0.0	10	0.0	75	100
Endpoints [mg/L] (based on nominal concentrations)
EC50		9.49 (4.42 – 21.15)
EC0		1.56
EC100		25
NOEC		6.25
LOEC		12.5

Validity criteria fulfilled:

yes

Remarks:

between 8.2 – 8.8 mg/L (not less than 3 mg/L)

Conclusions:

In the static Daphnia magna acute toxicity test the EC50 (48 h) is 9.49 mg/L, EC0 (48 h) value is 1.56 mg/L and EC100 (48 h) value is 25 mg/L - based on nominal concentrations of 2,4,6-trinitrotoluene (TNT). The NOEC value after 48 h is 6.25 mg/L. The LOEC value after 48 h is 12.5 mg/L.

Executive summary:

In a static acute toxicity study, young Daphnia magna individuals (< 24 h old) were exposed to the test item for 48 hours. Four replicates of each test concentration and control were tested with 5 daphnids per replicate (75 mL test volume). The test item 2,4,6-trinitrotoluene (TNT) was applied in six test concentrations: 0.39; 0.78; 1.56; 3.125; 6.25; 12.5 and 25 mg/L.

Daphnids were observed for immobilization after 24 and 48 hours of exposure. The animals are considered immobile if they showed no ability to swim within 15 seconds after shaking the test vessel.

The chemical analyses of test item 2,4,6-trinitrotoluene (TNT) performed by validated method confirmed correct preparation of test concentrations.

At test initiation the determined content of 2,4,6-trinitrotoluene (TNT) was in range of 98.3 -110.4%, of nominal concentrations. At test termination the analysed content of2,4,6-trinitrotoluene (TNT) was in range of 83.7 - 100.2 %of initial concentrations.

No immobilization of daphnids in test concentrations 0.39 – 1.56 mg/L, in concentration 6.25 mg/L and in control was observed. The test item caused immobilization of daphnids of 10; 75 and 100% in test concentrations 3.125; 12.5 and 25 mg/L, respectively.

Description of key information

In accordance with Regulation EC 1272/2008 (CLP) and basend on available data substance can be classified as hazardous to the aquatic environment chronic category 2 as 1 mg/L < LC50 < 10 mg/L.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Dose descriptor:

EC50

Effect concentration:

9.49 mg/L