1-nitroguanidine

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1993

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

test procedure in accordance with national standard methods with acceptable restrictions

Qualifier:

according to guideline

Guideline:

other: ASTM Designation E 729-80 (ASTM, 1980)

GLP compliance:

not specified

Specific details on test material used for the study:

Test substance supplier: U.S Naval Ordnance Station
Lot No: 985-1
Essay 99.998 %

Analytical monitoring:

yes

Vehicle:

no

Test organisms (species):

Pimephales promelas

Test type:

flow-through

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 3 320 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: No mortality occured in 96 h at the concentration shown which was the solubility limit of the compound in diluent water at 25 °C.

Validity criteria fulfilled:

not specified

Conclusions:

No mortality occurred to fathead minnow in 96 h at the solubility limit of the test compound at 3320 mg/L.
It can be concluded that the toxicity of nitroguanidine to fathead minnow is quite low. Nitroguanidine does not appear to be an acute hazard to freshwater fish.

Executive summary:

The 96 h acute toxicity of nitroguanidine Fathead minnow (Pimephales promelas) was studied under flow through conditions. Juveniles of Fathead minnow (10 per replicate) were exposed to measured concentrations of 0 (control), 470, 830, 1360, 2010, 3320 mg/l for 96 hours. The test system was maintained at 25.0 to 26.0 ºC and a pH of 7.8 to 8.5.

No mortality occurred to fathead minnow in 96 h at the solubility limit of the test compound at 3320 mg/L. The 96 h LC₅₀ for juveniles, based on mortality, was therefore > 3320 mg/L.

It can be concluded that the toxicity of nitroguanidine to fathead minnow is quite low. Nitroguanidine does not appear to be an acute hazard to freshwater fish.

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1993

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

test procedure in accordance with national standard methods with acceptable restrictions

Qualifier:

according to guideline

Guideline:

other: ASTM Designation E 729-80 (ASTM, 1980)

GLP compliance:

not specified

Specific details on test material used for the study:

Test substance supplier U.S Naval Ordnance Station
Lot No 985-1
Essay 99.998%

Analytical monitoring:

yes

Vehicle:

no

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Test type:

flow-through

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 1 550 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: No mortality occured in 96 h at the concentration shown which was the solubility limit of the compound in diluent water at 12 °C.

Validity criteria fulfilled:

not specified

Conclusions:

No mortality occurred to Rainbow trout in 96 h at the solubility limit at 12 °C of the test compound at 1550 mg/L.
It can be concluded that the toxicity of nitroguanidine to Rainbow trout is quite low. Nitroguanidine does not appear to be an acute hazard to freshwater fish.

Executive summary:

The 96 h acute toxicity of nitroguanidine Rainbow trout (Oncorhynchus mykiss) was studied under flow through conditions. Fingerlings of Rainbow trout (10 per replicate) were exposed to measured concentrations of 0 (control), 200, 370, 570, 950, and 1550 mg/L for 96 hours. The test system was maintained at 10.4 to 13.8 ºC and a pH of 7.4 to 7.7.

No mortality occurred to Rainbow trout in 96 h at the solubility limit of the test compound at 1550 mg/L (12 °C). The 96 h LC₅₀ for fingerlings, based on mortality, was therefore > 1550 mg/L.

It can be concluded that the toxicity of nitroguanidine to Rainbow trout is quite low. Nitroguanidine does not appear to be an acute hazard to freshwater fish.

Reference 3

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1985

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

test procedure in accordance with national standard methods with acceptable restrictions

Qualifier:

according to guideline

Guideline:

other: Am. Soc. Test. Mat. 1980. Standard practice for conducting acute toxicity tests with fishes, macroinvertebrates, and amphibians. E 729-80.

Deviations:

not specified

GLP compliance:

not specified

Specific details on test material used for the study:

Test substance supplier: Aldrich Chemical Co
Water Content: 25 %

Analytical monitoring:

yes

Vehicle:

no

Test organisms (species):

other: 4 test species: Pimephales promelas, Lepomis macrochirus, Ictalurus punctatus, and Salmo gairdneri

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 2 714 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: result for nitroguanidine solution on fathead minnow

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

34.5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

photolyzed nitroguanidine

Basis for effect:

mortality (fish)

Remarks on result:

other: effect of photolyzed nitroguanidine solution on fathead minnow

Sublethal observations / clinical signs:

- Except in the bluegill static acute test, control mortality never exceeded 3 percent.

- Rainbow trout was the most sensitive fish species.

- LC50 (96h; nitroguanidine) values for all fish species were greater than the saturation concentration at the used test temperature.

Table 1: Acute toxicity of nitroguanidine to fish 

Species	Length of exposure (h)	Mean measured concentration^a (mg/L)	Percent mortality
Rainbow trout	96	1638	20
		901	0
		< 1.45	0
Fathead Minnow	96	2714	0
		1690	0
		< 1.45	0
Bluegill	96	2634	0
		1671	0
		< 1.45	13.3
Channel Catfish	96	2636	0
		1644	0
		< 1.45	0

^athe high, low and control nitroguanidine concentrations are listed

 

Table 2: The effect of photolyzation on the acute toxicity of nitroguanidine to Fathead Minnow

Species	Endpoint	Toxicant concentration (mg/L)		Relative Toxicity Ratio, photol. NGu/ NGu
		NGu	Photolyzed NGu^a
Fathead Minnow	LC50(96h)	> 2.714^b	34.5 (22.1-61-3)	> 78

^aConcentration reported as NGu (mg/l) prior to photolyzation. No mortality was observed in the photolyzed well water control tanks

^bNo mortality at 2714 mg/L

 

Validity criteria fulfilled:

not specified

Conclusions:

The toxicity of nitroguanidine to the freshwater fish was uniformly low.
Based on these results, nitroguanidine appears to present little hazard to aquatic organisms.

Executive summary:

The 96 h acute toxicity of nitroguanidine to fish (4 test species: Pimephales promelas, Lepomis macrochirus, Ictalurus punctatus, and Salmo gairdneri) was studied under static conditions. Fish were exposed to nitroguanidine at two concentrations (nitroguanidine saturation concentration at test temperature and 50 % of saturation). In addition fathead minnow (P. promelas) was exposed to photolyzed nitroguanidine at least at four test concentrations so that an LC₅₀ could be adequately defined.

The LC₅₀(96h) for nitroguanidine for all fish species were greater than the saturation concentration at the used test temperature. 

The LC₅₀(96h) for fathead minnow (P. promelas) for photolyzed nitroguanidine was 34.5 mg/L (mean measured concentration).

 

Based on the results of this study, nitroguanidine appears to present little hazard to fish. Photolysis increased the acute toxicity in fathead minnow by a factor from over 78.

Reference 4

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

April 2000

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

GLP compliance:

not specified

Analytical monitoring:

no

Details on sampling:

not applicable

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- A stock solution of 1600 mg/L moistened test material in tap water was prepeared.
- From this stock solution the final nominal test concentrations were prepared by dillution with tap water.
- Controls: test medium without test substance

Test organisms (species):

Poecilia reticulata

Details on test organisms:

TEST ORGANISM
- Common name: Guppy
- Source: laboratory breeding:
* parental and young fish population were held in 250 L glass aquaria
* illumination: 14h/d
* temperature: 23 +/- 2 °C
* medium: continously aerated and filtered tap water from Schlieren; the bottom of the aquaria was covered with gravel (≤ 1.5 cm); every
month, one third of the total volume of the aquaria was replaced by fresh tap water
* feed: Sera micropan, Sera vipan daily; the fish were last fed 24 h before the start of the definitive test
* measurements: pH, conductivity, O2, and hardness of water were determined once a week; temperature was controlled by a thermostatic
heating element
- Length at study initiation: 1-3 cm
- Weight at study initiation (mean and range, SD):
- Feeding during test: no

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

no data

Test temperature:

21 +/- 1 °C

pH:

8.3-8.9

Dissolved oxygen:

7.0-8.1

Salinity:

no data

Nominal and measured concentrations:

- 0 (control), 100, 200, 400, 800, 1600 mg/L (80 % active ingredient!)
- no analytical verification of test concentrations

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type: open, 1 L aquaria, all-glass
- Aeration: no
- Type of flow-through (e.g. peristaltic or proportional diluter):
- Renewal rate of test solution (frequency/flow rate):
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1



TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: tap water from Schlieren, aerated
- Total hardness: 2.4-4.1 °F
- Chloride: 7.5-12.0 mg/l
- Sulfate: 17.5-21.0 mg/l
- PO4-P: < 0.002-0.038 mg/l
- NH4-N: < 0.01 mg/l
- NO2-N: < 0.002 mg/l
- NO3-N: 7.0-12.0 mg/l
- Conductivity: 510-585 µS/cm
- Culture medium different from test medium: no
- Intervals of water quality measurement: no data


OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 14 h
- Light: overhead white fluorescent tubes


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
observations of the fish were made after 2, 24, 48, 72, and 96 h of exposure; mortality of the fish was recorded at the same time intervals


TEST CONCENTRATIONS
- Range finding study
- Test concentrations: 1, 10, and 100 mg/l
- three animals per test vessel
- Results used to determine the conditions for the definitive study: 1 of 3 fish died at 10 mg/L after 96 h of exposure; no mortality was observed
at 100 and 1 mg/L and in the control after 96 h of exposure

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 1 600 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

80% active ingredient

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

>= 800 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

80% active ingredient

Basis for effect:

behaviour

Details on results:

- Behavioural abnormalities: loss of coordination, hypoactivity and swimming on the back were observed at 1600 mg/L
- In the definitive test 1 of 7 fish died at the nominal concentration of 100 mg/l after 96 h of exposure; no mortality was observed in all other test concentrations and in the control after 96 h of exposure

Results with reference substance (positive control):

Not applicable

Reported statistics and error estimates:

No statistics reported

Sublethal observations / clinical signs:

- at least 1 h before the test, the test vessels were filled with the medium to allow equilibration of pH, oxygen and temperature

- to start the test fish were transferred into the test vessels about 1h after formation of the test media

Table 1: Incidence of mortality observed

Nominal concentration (mg/l)	Number of dead fish observed at different points in time after start of exposure
	2 h	24 h	48 h	72 h	96 h
0, Control	0	0	0	0	0
100	0	0	0	0	1
200	0	0	0	0	0
400	0	0	0	0	0
800	0	0	0	0	0
1600	0	0	0	0	0

 

Table 2: Specification of symptoms observed on affected fish and data of dead fish

Nominal concentration (mg/l)	Number of affected fish^a)	Time of score (h)	Body-length (mm)	Body-weight (mg)	Symptom^b)
1600	7/7	24	-	-	hypoactivity
1600	1/7	48	-	-	hypoactivity
1600	1/7	72	-	-	hypoactivity
100	1/6	96	12	17	dead, at bottom

a) Number of affected fish/number of surviving fish

b) e.g. hyper-/hypoactivity, immobility, swimming at the bottom/surface, darkened pigmentation/discoloring

Validity criteria fulfilled:

yes

Remarks:

During 7 days before the definitive test, mortality in the batch of which fish were used for the test was less than 5 %.

Conclusions:

Based on these data the nominal lethal concentration (LC50) of nitroguanidin (moistened; 80% active ingredient) to Poecilia reticulata was estimated to be > 1600 mg/L.
The NOEC was ≥ 800 mg/L since the one fish that died at 100 mg/L is considered not to represent a significant effect as defined by the validity criteria of the test.
Therefore, it can be concluded that nitroguanidin is practically non-toxic towards Poecilia reticulata under the conditions of this test.

Executive summary:

In a static 96 h acute toxicity study, Poecilia reticulata (Guppy) were exposed to nitroguanidine (moistened; ~ 80 % active ingredient) at nominal concentrations of 0 (control), 100, 200, 400, 800 and 1600 mg/L under static conditions.

The 96 h LC₅₀was > 1600 mg/L (equivalent to > 1280 mg/L active ingredient).  NOEC value, based on mortality, was ≥ 800 mg/L (equivalent to ≥ 640 mg/L active ingredient). Sublethal effects (loss of coordination, hypoactivity and swimming on the back) were observed at 1600 mg/L (equivalent to 1280 mg/L active ingredient).

This toxicity study is classified as acceptable and satisfies the OECD 203 guideline requirements.

Description of key information

Acute (96 h) fish toxicity test with nitroguanidine were conducted with several fish species (Poecilia reticulata, Pimephales promelas, Lepomis macrochirus, Ictalurus punctatus, and Salmo gairdneri; see Häner & Höin (2000), van der Schalie (1985), and Burton et al. (1993)) at concentrations up to 3320 mg/L. In addition, van der Schalie conducted experiments with photolyzed nitroguanidine (four initial nitroguanidine concentrations). Häner & Hoin (2000), van der Schalie (1985) and Burton et al. (1993) determined LC50 values to be higher than 1280 mg/L, 2714 mg/L and 1550 mg/L, respectively. From these results it can be concluded that nitroguanidine is not of acute toxicity to fish species. In contrast, the LC50 for photolyzed nitroguanidine determined by van der Schalie (1985) is 34.5 mg/L being two orders of magnitude more toxic than nitroguanidine.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Dose descriptor:

LC50

Effect concentration:

34.5 mg/L

Additional information

In a 96 h acute toxicity study, Häner & Höin, 2000, exposed Poecilia reticulata (Guppy) to nitroguanidine (moistened; ~ 80 % active ingredient) at nominal concentrations of 0 (control), 100, 200, 400, 800 and 1600 mg/L under static conditions. The 96 h LC₅₀ to be > 1600 mg/L (equivalent to > 1280 mg/L active ingredient).  The NOEC value, based on mortality, was 800 mg/L (equivalent to 640 mg/L active ingredient).  Sublethal effects (loss of coordination, hypoactivity and swimming on the back) were observed at 1600 mg/L (equivalent to 1280 mg/L active ingredient).

In studies conducted by van der Schalie, 1985, the 96 h acute toxicity of nitroguanidine to fish (4 test species: Pimephales promelas, Lepomis macrochirus, Ictalurus punctatus, and Salmo gairdneri) was studied under static conditions. Fish were exposed to Nitroguanidine at two concentrations (nitroguanidine saturation concentration at the test temperature and 50 % of saturation). In addition fathead minnow (P. promelas) was exposed to photolyzed nitroguanidine at least at four test concentrations so that an LC50 could be adequately defined. In the tests conducted by van der Schalie, 1985, the LC₅₀ (96 h) for nitroguanidine for all fish species were greater than the saturation concentration at the test temperature used. Rainbow trout was the most sensitive fish species displaying the highest mortality (20 %) at the maximum concentration (1638 mg/l; mean measured concentration) at the test temperature used.

The LC50 (96 h) for fathead minnow (P. promelas) for photolyzed nitroguanidine was 34.5 mg/l (mean measured concentration).

Burton et al., 1993, determined the 96 h acute toxicity of nitroguanidine Fathead minnow (P. promelas) under flow through conditions. Juveniles of Fathead minnow (10 per replicate) were exposed to measured concentrations of 0 (control), 470, 830, 1360, 2010, 3320 mg/l for 96 hours. The test system was maintained at 25.0 to 26.0 ºC and a pH of 7.8 to 8.5.

Supporting the results of van der Schalie, 1985, Burton et al., 1993, observed no mortality to fathead minnow in 96 h at the solubility limit of the test compound at 3320 mg/L. The 96 h LC₅₀ for juveniles, based on mortality, was therefore > 3320 mg/L.

From these results it can be concluded that the toxicity of nitroguanidine to freshwater fish is quite low. Nitroguanidine does not appear to be an acute hazard to freshwater fish.

 

Table 1: Study synopsis

Guideline Test Type	Species	Effect Concentration	Reference	Reliability
OECD 203	Poecilia reticulata	LC50(96h) mg/L > 1600 (equivalent to > 1280 mg/L active ingredient)   NOEC = 800 mg/L (equivalent to 640 mg/L active ingredient)	Häner, A.; Höin, C.; BMG Engineering AG; 2000	Reliable without restrictions
m. Soc. Test. Mat. 1980. Standard practice for conducting acute toxicity tests with fishes, macroinvertebrates, and amphibians. E 729-80.	Pimephales promelas	Nitroguanidine: LC50(96h) mg/L > 2714   Photolyzed Nitroguanidine: LC50(96h) mg/L = 34.5	Schalie, W.H. van der; U.S.Army Medical Bioengineering Research and Development Laboratory; 1985	Reliable with restrictions
ASTM Designation E 729-80 (ASTM, 1980)	1.  Oncorhynchus mykiss     2.  Pimephales promelas	LC50(96h) mg/L > 1550      LC50(96h) mg/L > 3320	Burton, D.T.; Turley, S.D.; Peters, G.T.; University of Maryland; 1993	Reliable with restrictions