Tricarbonyl(methylcyclopentadienyl)manganese

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:

1990-02-27 until 1990-03-01

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted according to EPA OTS 797.1300 (Aquatic Invertebrate Acute Toxicity Test, Freshwater Daphnids) and following GLP.

Qualifier:

according to guideline

Guideline:

EPA OTS 797.1300 (Aquatic Invertebrate Acute Toxicity Test, Freshwater Daphnids)

Deviations:

no

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes (incl. QA statement)

Remarks:

Statement of compliance with U.S. EPA Good Laboratory Practice Standards; Toxic Substances Control (40 CFR 792)

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
No further details presented

Analytical monitoring:

yes

Details on sampling:

- Concentrations:
Nominal concentrations: Control (Acetone), Solvent control, 0.65, 1.3, 2.5, 5.0 and 10 mg/L
Analytical concentrations: Control (Not detected), Solvent control (Not detected), 0.41, 0.91, 1.3, 2.6, 4.1 mg/l (0-hours) and 0.17, 0.39, 0.70, 1.4, 2.8 mg/l (48-hours). Mean measured concentration (0 and 24 hours) 0.29, 0.65, 1.0, 2.0 and 3.5 mg/l.
- Sampling method: At the 0-hour sampling period, the 50 and 100 ml sample aliquots were collected directly from the volumetric flasks in which the test solutions were prepared. At the 48-hour sampling, 25 ml and 50 ml aliquots were composited from replicates A and B of each test concentration.
- Sample storage conditions before analysis: The samples were not stored but extracted immediately and analysed within 30 minutes after sampling.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: AQpropriate aliquots were taken from a primary mmtstandard of 100 mg/l and added to a 1 liter volumetric containing test water. The test solutions were poured from the volumetric into the appropriately labeled test vessels.
- Eluate: Test water was a blend of ABC well water and soft blended water. Certificate of regular analysis of the water is present in the report.
- No vehicle
- Controls: Blank and solvent control
- Evidence of undissolved material: None

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Source: ABC laboratories: in house culture.
- Age at study initiation (mean and range, SD): <24 hours old (neonates)
- Weight at study initiation (mean and range, SD): Not relevant
- Length at study initiation (length definition, mean, range and SD): Not relevant
- Valve height at study initiation, for shell deposition study (mean and range, SD): Not relevant
- Peripheral shell growth removed prior to test initiation: No relevant
- Method of breeding: The Daphnia magna used in the test were cultured at the ABC facilities in hard blended water. All daphnids were held in a temperature controlled area at 20±2 ºC. The Daphnids were fed algae at least every three days prior to testing and supplemented with a suspension of trout chow and yeast (in accordance with OECD 202 guidelines)
- Feeding during test: Animals were not fed during the 48-hours test period.


ACCLIMATION
- Acclimation period: The animals were an in-house culture. Acclimation not required.
- Acclimation conditions (same as test or not): 20°C and a photoperiod of 16 hrs light and 8 hrs dark.
- Type and amount of food: Original culture of Daphnia : algae Selenastrum capricornutum and supplemented with a suspension of trout chow and yeast.
- Feeding frequency: every three days
- Health during acclimation (any mortality observed): No

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Post exposure observation period:

No.

Hardness:

172 mg/L as CaCo3

Test temperature:

20°C

pH:

7.9 to 8.3

Dissolved oxygen:

7.5-8.2 mg/L

Salinity:

No

Nominal and measured concentrations:

- Concentrations:
Nominal concentrations: Control, Solvent control, 0.65, 1.3, 2.5, 5.0 and 10 mg/L f test material.
Analytical concentrations: Control (Not detected), Solvent control (Not detected), 0.41, 0.91, 1.3, 2.6, 4.1 mg/l of test material (0-hours) and 0.17, 0.39, 0.70, 1.4, 2.8 mg/l test material (48-hours). Mean measured concentration (0 and 24 hours) 0.29, 0.65, 1.0, 2.0 and 3.5 mg/l test material.

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type: closed with loose-fitting petri-dish covers
- Material, size, headspace, fill volume: 250 ml glass beakers filled with 200 ml of test volume
- Aeration: None
- Type of flow-through: Not relevant.
- Renewal rate of test solution: Not relevant.
- No. of organisms per vessel: 10 daphnids
- No. of vessels per concentration : 2
- No. of vessels per control (Blank) : 2
- No. of vessels per control (Solvent control) : 2
- No. of vessels per vehicle control (replicates): No vehicle control was used
- Biomass loading rate: 10 daphnids/200 ml


TEST MEDIUM / WATER PARAMETERS
- Total organic carbon: <1 ppm
- Particulate matter: 1.4 ppm
- Metals: Aluminum: <0.20 ppm; Arsenic: <1.0 ppb; Boron: 0.318 ppm; Cadmium: <0.005; Chromium: <0.001 ppm; Cobalt: <0.050 ppm; Copper: <0. 025 ppm; Fluoride: 0.40 ppm; Iron: <0.10 ppm; Lead: <1.0 ppb; Mercury: <0.0004 ppm; Nickel: <0.040 ppm; Silver: <1.0 ppb; Zinc: <0.033 ppm.
- Pesticides: DDE: <0.05 ppb; DDD: <0.05ppb; DDT: <0.05ppb; Dieldrin: <0.05ppb; α-BHC: <0.01 ppb; β-BHC: <0.01 ppb; γ-BHC: <0.01 ppb; Δ-BHC: <0.01 ppb; HCB: < 0.01 ppb; Endrin: <0.05 ppb; H.E.: <0.01 ppb; Mirex: <0.10; Methoxyclor: <0.10; Toxaphene: <1.0 ppb
- Chlorine: No
- Alkalinity: 192 mg/L as CaCO3
- Conductivity: 325 µMhos/cm
- Culture medium different from test medium: No
- Intervals of water quality measurement: Water chemistry parameters of temperature, dissolved oxygen and pH were performed on all test concentrations at 0-hour and 48-hours.


OTHER TEST CONDITIONS
- Adjustment of pH: None reported
- Photoperiod: 16 hrs light/8 hrs dark
- Light intensity: 50-70 foot candles


EFFECT PARAMETERS MEASURED : All daphnids in each test vessel were observed once at 4-, 24- and 48 hours for immobility and abnormal effects such as surfacing, clumping of the daphnids together and daphnids tending to the bottom of the test chambers


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2
- Range finding study: Two range-finding studies were performed to determine the adequate concentrations to be used in the study test. Since the concentrations chosen in the first test study did not had a sufficient response (Only 10% immobility at the highest concentration) , a second range-finding study was conducted.
- Test concentrations: First range-finding study: 0.001, 0.01, 0.10 and 1.0 mg/L; Second range-finding study: 1.0 and 10.0 mg/L (90% immobility was observed at concentration of 10 mg/L)
- Results used to determine the conditions for the definitive study: Since no immobility was observed at 1.0 mg/L and 90% immobility was observed at 10.0 mg/L, the range test of the second study was established based on the results.

Reference substance (positive control):

no

Duration:

24 h

Dose descriptor:

EC50

Remarks:

Binomial method

Effect conc.:

0.94 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95% CL 0.78 and 1.1 mg/l

Duration:

48 h

Dose descriptor:

EC50

Remarks:

Binomial Method

Effect conc.:

0.83 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95% CL 0.70 and 0.99 mg/l

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

0.29 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- Behavioural abnormalities: Surfacing was observed at the concentration of 0.65 mg/L of mmt, 1 daphnid of vessel A and 3 daphnids of vessel B already at 4-hours exposure. Detailed results on immobilization data presented on "Remarks on results including tables and figures"
- Observations on body length and weight: Not required.
- Other biological observations: Daphnids exhibited surfacing and clumping together. They tended to the bottom of test chambers
- Mortality of control: None
- Other adverse effects control:None
- Abnormal responses: None
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: None
- Effect concentrations exceeding solubility of substance in test medium: None

Results with reference substance (positive control):

No reference substance was used.

Individual immobility and behavioral observations

for the Acute Toxicity Test of mmt to Daphnia magna

Mean measured Concentration (mg/L)	Rep.	Number placed in test vessel	4-hour		24-hour		48-hour
			Cum. Immob.	Obs.	Cum. Immob.	Obs.	Cum. Immob.	Obs.
Control	A	10	0	10N	0	10N	0	10N
	B	10	0	10N	0	10N	0	10N
Solvent control	A	10	0	10N	0	10N	0	10N
	B	10	0	10N	0	10N	0	10N
0.29	A	10	0	10N	0	10N	0	10N
	B	10	0	10N	0	10N	0	10N
0.65	A	10	0	1 SUR* 9N	1	9N	2	8N
	B	10	0	3 SUR* 7N	2	8N	3	7N
1.0	A	10	8^b	2N	8	2N	8	2N
	B	10	8^b	2N	6	4N	7	3N
2.0	A	10	9	1N	9	1N	9	1N
	B	10	9	1N	9	1N	10	----
3.5	A	10	10	----	10	----	10	----
	B	10	10	----	10	----	10	----

^aUnless otherwise indicated, the test water was clear and free of precipitate. The following abbreviations were used for observations: SUR=Surfacing, N=.normal

^bOn the surface.

* Pushed back under the surface to insure that surfacing was a compound related effect and not just air trapped under the daphnids carapace during addition to the test vessel.

Validity criteria fulfilled:

yes

Remarks:

Validity criteria were fulfilled. No significant immobilization of daphnids was observed in the control (less then 10%). Oxygen concentration > 3.0 mg/l

Conclusions:

Test results showed that mmt has adverse effects on Daphnids at concentrations up to 10.0mg/L. The EC50 calculated at 48-hous was 0.83 mg/L using the binomial method with a 95% CL 0.70 and 0.99 mg/l.

Executive summary:

The 48-hours acute toxicity of mmt to Daphnia magna was studied under static system conditions. Daphnids were exposed to controls, solvent controls and the test material at nominal concentrations of 0.65, 1.3, 2.5, 5.0 and 10 mg/L and average analytical concentrations of 0.29, 0.65, 1.0, 2.0 and 3.5 mg/L. The measured values were lower than the nominal concentrations as mmt is photosensitive and degrades when exposed to the light.. The calculated toxicity values were, based on the geometric mean measured concentrations as they were considered the best estimate of toxicity of MMT to daphnia magna.

 

The 48-hours EC50 was determined to be 0.83 mg/L using the binomial method with 95% CL of 0.70 and 0.99 mg/l. The 48 -hour NOEC was determined to be 0.029 mg/. This study is classified as acceptable and satisfies the guideline requirements for an acute toxicity study with Daphnia magna.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

0.83 mg/L