Hexahydro-4-methylphthalic anhydride

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 January 25, 2010 to February 03, 2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Polynt Lot No. T408209159
- Manufacturing date: 08 Jun 2009
- Expiration date of the lot/batch: 08 Jun 2010

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Keep container tightly closed. Keep in a dry, cool and well ventilated place.
- Stability under test conditions: The test material reacts forming the corresponding acid: 1,2-Cyclohexanedicarboxylic acid, 4-methyl- (in water and light at room temperature).
- Storage at testing facility: 5 years

Analytical monitoring:

yes

Details on sampling:

Analytical verification (0 and 96 hours)

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: A static limit test was performed with one application at experimental starting
- Controls: 7 fish in dilution water (without test item) were tested under the same test conditions as the test replicates.
- Concentration : 100 mg/L
- The test medium was mixed with an ultraturrax (1 min, 7000 rpm )
- Test volume : About 20 L per vessel.

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Common name: Gnathostoma, Pisces, Osteichthyes, Teleostei, Salmoniformes, Salmonidae
- Strain: Oncorhynchus mykiss (Rainbow trout)
- Source:
For preliminary test: Forellenzucht Kasselmann, Forellental 12, D-49170 Hagen
For definitive test: Forellenzucht Trostadt GbR, Dorfstr. 7, D-98646 Trostadt
- Age at study initiation (mean and range, SD):
- Length at study initiation (length definition, mean, range and SD): 5.67 cm
- Weight at study initiation : 2.00 g


ACCLIMATION
- Acclimation period: At least 12 days.
- Acclimation conditions (same as test or not): Holding was performed at the test facility at 13 - 17 °C, diffuse light (0.1-10 µmol photons .m-2 . s -1, 12 h photoperiod daily) and under flow-through conditions. The water exchange was 1-2 times the water volume per day.
- Type and amount of food: during acclimatisation food was provided 3 times per week. The test fish were not fed 24 hours before test start. Food type: Aller Performa Gr. 2 (ALLER AQUA A/S, Allervej 130, DK-6070 Christiansfeld). Amount: The amount of food was 4 % of the fish body weight per feeding day.
- Health during acclimatisation (any mortality observed): No mortality was observed during this time. No disease treatments were administered throughout holding and testing.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

10 - 250 mg CaCO3 / L

Test temperature:

13-17 °C

pH:

6.0 - 8.5

Dissolved oxygen:

The dissolved oxygen concentration was more than 80 % of the air saturation value.

Nominal and measured concentrations:

Sampling date 2010-01-25 2010-01-29
Start of analysis 2010-01-25 2010-02-03

Nominal concentration Hexahydro - 4 Methylphthalic Anhydride

Test item [mg/L] Measured conc. RR Measured conc. RR
[mg/L] [%] [mg/L] [%]

100 93.7 94 109 109
100 pH-control 97.5 98 107 107
Control < LOQ < LOQ

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass tank loosely covered by glass tops were used.
- Material, size, headspace, fill volume: 20 Lt per vessel
- Aeration: Gentle aeration was provided
- No. of organisms per vessel: Fish density in the tanks was < 1 g fish per litre test solution.
- No. of vessels per concentration (replicates): one vessel with 7 Rainbow trout for the test ITEM , one vessel with 7 Rainbow trout where the pH is
adjusted
- No. of vessels per control (replicates): One vessel with 7 Rainbow trout

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Tap water of local origin was used for holding and testing. The water was filtered on activated charcoal and aerated for at least 24 hours to remove chlorine.
- pH-value: 6.0 - 8.5
- Hardness: 10 - 250 mg CaCO3 / L


OTHER TEST CONDITIONS
- In one vessel the pH-value of the test medium was adjusted to the pH of the dilution water ± 0.2 by adding appropriate volumes of 7 mol/L NaOH.
- Photoperiod: A daily 12 hours photoperiod occurred during the test.
- Light intensity: 0.1 -10 µmol photons . m-2 . s-1
- Feeding of the test fish: The fish were not fed during the test.

Reference substance (positive control):

no

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

The limit test concentration caused no mortality or non-lethal effects in both test vessels (with and without adjustment of the pH-value) at a concentration of 100 mg/L.

Sublethal observations / clinical signs:

Cumulative Mortality [%] in the Test Vessels

(n=7)

Nominal test item concentration [mg/L]	Test Duration [hours]
	2 – 4	24	48	72	96
100	0	0	0	0	0
100 (pH-control)	0	0	0	0	0
Control	0	0	0	0	0

Validity criteria fulfilled:

yes

Conclusions:

In this study Hexahydro - 4 Methylphthalic Anhydride (4-MHHPA) did not cause any effects to Rainbow trout after 96 hours tested with the limit concentration of 100 mg/L test item, with and without adjustment of the pH-value.

Executive summary:

Acute toxicity to fish of 4 -MHHPA has been investigated in accordance with OECD/EU test methods. A static limit test was conducted with a single test item concentration of 100 mg/L with and without adjustment of the pH-value. No mortality occurred over the 96 hour duration of the test.

Description of key information

Acute toxicity to fish: LC50 = > 100 mg/L; NOEC = 100 mg/L

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

100 mg/L

Additional information

Acute toxicity to fish of 4-MHHPA has been investigated in accordance with OECD/EU test methods. A static limit test was conducted with a single test item concentration of 100 mg/L with and without adjustment of the pH-value. No mortality occurred over the 96 hour duration of the test. These findings are supported by a test on the acid degradation product in which the 48 hour LC50 was determined to be >= 500 mg/L.