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REACH

Reaction mass of 4-(2,6,6-trimethylcyclohex-2-ene-1-yl)-but-3-ene-2-one and 4-(2,6,6-trimethylcyclohex-1-ene-1-yl)-but-3-ene-2-one

EC number: 907-706-6 | CAS number: -

Life Cycle description
Uses advised against

Ecotoxicological information

Long-term toxicity to aquatic invertebrates

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Administrative data

Link to relevant study record(s)

Reference

Endpoint:: long-term toxicity to aquatic invertebrates
Type of information:: experimental study
Adequacy of study:: weight of evidence
Reliability:: 2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:: guideline study
Justification for type of information:: Data is from experimental study report
Qualifier:: according to guideline
Guideline:: OECD Guideline 211 (Daphnia magna Reproduction Test)
GLP compliance:: not specified
Analytical monitoring:: yes
Details on sampling:: The stock solution was prepared by dissolving 100 mg of test chemical in 100 ml of M7 medium with continuous 24 hours stirringto get the final concentration of 1000 mg/L and this stock solution was filtered with whatman filter paper No. 41 followed by analytical determination. The final solubility* value obtained in media was used to prepare the remaining test concentrations from the above stock solution.
Vehicle:: no
Details on test solutions:: PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
Test chemical was weighed and about 100mg in 100 ml of test chemical was dissolved in M7 medium and kept for stirring for 24 hours, which was then filtered and the concentration was anlytically determined.
- Controls: Test media M7 was directly used as the control.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): no vehilce was used during study
- Concentration of vehicle in test medium (stock solution and final test solution(s) or suspension(s) including control(s)): not applicable
Test organisms (species):: Daphnia magna
Details on test organisms:: TEST ORGANISM
- Common name: Water flea
- Age at study initiation (mean and range, SD): < 24 hr old
- Length at study initiation (length definition, mean, range and SD): 0.45 cm
- Source: Eggs of Daphnia magna were obtained from Denmark Technical University and maintained at Unique Ecotox Research Laboratory, Nagpur
- Stage and instar at study initiation: Neonates <24 hours
- Method of breeding: Gravid daphina were sperated and maintained in the fresh media, the neonates obatined from the second brood pouch was taken for the study.
- Source: Eggs of Daphnia magna were obtained from Denmark Technical University and maintained in laboratry
- Age of parental stock (mean and range, SD): >4 weeks
- Feeding during test: feeding was given during the stiudy thrice a week
- Food type: Selenestrum capricornutum
- Frequency: thrice a week

ACCLIMATION
- Type and amount of food: Living cells of Selenestrum capricornutum
- Feeding frequency: Feeding of the Daphnia with living cells of Selenestrum capricornutum was done thrice a week.
Test type:: semi-static
Water media type:: freshwater
Limit test:: no
Total exposure duration:: 21 d
Hardness:: 140 mg/l as CaCO3
Test temperature:: 18-22°C
pH:: 7.4-7.9
Dissolved oxygen:: 7.4-8.1mg/L
Salinity:: Not applicable
Nominal and measured concentrations:: 0.175, 0.228, 0.296, 0.385, and 0.5mg/L
Details on test conditions:: TEST SYSTEM
- Test vessel: glass beaker
- Material, size, headspace, fill volume: 100 ml, fill volume 50 ml, and head space 50ml
- Aeration: Test vessel were not aerated during the study.
- No. of vessels per concentration (replicates):10
- No. of vessels per control (replicates): 10
- No. of vessels per vehicle control (replicates): not appliicable
- Renewal rate of test solution (frequency/flow rate): renewed every 24 hours

TEST MEDIUM / WATER PARAMETERS: M7 medium

OTHER TEST CONDITIONS
- Photoperiod: 16:8 light:dark conditions
- Light intensity: Not exceeding 1000-1500 lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : The evaluation of the NOEC was determined based on the number of offspring’s produced per living parent Daphnia.
Reference substance (positive control):: not required
Duration:: 21 d
Dose descriptor:: NOEC
Effect conc.:: 0.175 mg/L
Nominal / measured:: nominal
Conc. based on:: test mat.
Basis for effect:: reproduction
Duration:: 21 d
Dose descriptor:: LOEC
Effect conc.:: 0.228 mg/L
Nominal / measured:: nominal
Conc. based on:: test mat.
Basis for effect:: reproduction
Details on results:: - Behavioural abnormalities: no behavioural abnormalities were observed
- Observations on body length and weight: no significant difference was observed in the body length was observed in both parents and offsprings
- Other biological observations: first brood with visible eggs in parents was observed on day 11
- Mortality of control: no moratality was observed in control parents and offsprings
- Other adverse effects control: no adverse effects were observed
- Immobilisation of control: no immobilisation
Results with reference substance (positive control):: Not applicable
Reported statistics and error estimates:: The statistical comparision was done by student T test method using Sys stat software
Validity criteria fulfilled:: yes
Conclusions:: Based on the effect on reproduction of the test daphnids, the 21 d NOEC qnd LOEC value was determined to be 0.175 and 0.228 mg/l.
Executive summary:: A chronic study was conducted for assesing the effect of test chemical. The study was performed in accordance with the principles of the OECD Guideline 211 (Daphnia magna Reproduction Test). Daphnia magna of length 0.45 cm was used as a test organism. Eggs of Daphnia magna were obtained from Denmark Technical University. Test daphnids were fed with living cells of Selenestrum capricornutum and it was done thrice a week. Therefore, the test chemical will be prepared by dissolving 100 mg of test chemical in 100 mL of M7 media with 48 hours stirring to get the final concentration of 1000 mg/L. This stock solution will be filtered by using whatman filter paper no. 42, which will then analytically determine. The final solubility value obtained in media was used to prepare the remaining test concentrations from the above stock solution. The analytical determinations were performed by UV-VIS spectrophotometer. The pre-treated stock solution was then diluted with media in order to get the required test solutions ranging from concentrations, 1, 2, 3,4,5, 6,7, 8, 9 and10mg/L. The absorbance of resulting solution was measured using UV-VIS spectrophotometer against corresponding blank at lambda max (λmax). Standard curve was plotted against concentration verses absorbance and the maximum solubility was determined from the below standard curve. Analytical assessments were performed for selected test concentration at 1st, 2nd and 3rd week. The concentration of the test chemical being tested has been satisfactorily maintained within ± 20 % of the nominal concentration throughout the test. Therefore, the analysis of the results was based on nominal concentration. Test chemical conc. used in the defiinite study was 0.175, 0.228, 0.296, 0.385, and 0.5mg/Lmg/l. Thus, limit test was performed at 0.1 mg/l conc. Test daphnids were exposed to test chemical conc. in a glass beaker for an exposure period of 21 days. Vessels were not aerated during the study. M7 medium was used as a test medium. Test conditions involve a temperature range of 18-22°C, 16:8 light:dark conditions and light intensity not exceeding 1000-1500 lux, respectively. The evaluation of the NOEC was determined based on the number of offspring’s produced per living parent Daphnia. Defined concentrations of the test chemical led to a certain percentage reduction of the parthenogenetic reproduction rate at the end of the 21 day study period. The living offspring was counted daily along with the renewal of the test medium. However, test vessels were inspected daily for the occurrence of juveniles and marked accordingly. On the basis of the effect on reproduction of the test daphnids, the 21 d NOEC and LOEC values was determined to be 0.175 and 0.228mg/l. Thus based on the outcomes chemical could be classified into aquatic chronic category 3 as per CLP classification criteria.

Description of key information

Long term toxicity to aquatic invertebrates:

A chronic study was conducted for assesing the effect of test chemical. The study was performed in accordance with the principles of the OECD Guideline 211 (Daphnia magna Reproduction Test). Daphnia magna of length 0.45 cm was used as a test organism. Eggs of Daphnia magna were obtained from Denmark Technical University. Test daphnids were fed with living cells of Selenestrum capricornutum and it was done thrice a week. Therefore, the test chemical will be prepared by dissolving 100 mg of test chemical in 100 mL of M7 media with 48 hours stirring to get the final concentration of 1000 mg/L. This stock solution will be filtered by using whatman filter paper no. 42, which will then analytically determine. The final solubility value obtained in media was used to prepare the remaining test concentrations from the above stock solution. The analytical determinations were performed by UV-VIS spectrophotometer. The pre-treated stock solution was then diluted with media in order to get the required test solutions ranging from concentrations,1, 2, 3,4,5, 6,7, 8, 9 and10mg/L. The absorbance of resulting solution was measured using UV-VIS spectrophotometer against corresponding blank at lambda max (λmax). Standard curve was plotted against concentration verses absorbance and the maximum solubility was determined from the below standard curve. Analytical assessments were performed for selected test concentration at 1st, 2nd and 3rd week. The concentration of the test chemical being tested has been satisfactorily maintained within ± 20 % of the nominal concentration throughout the test. Therefore, the analysis of the results was based on nominal concentration. Test chemical conc. used in the defiinite study was 0.175, 0.228, 0.296, 0.385, and0.5mg/Lmg/l. Thus, limit test was performed at 0.1 mg/l conc. Test daphnids were exposed to test chemical conc. in a glass beaker for an exposure period of 21 days. Vessels were not aerated during the study. M7 medium was used as a test medium. Test conditions involve a temperature range of 18-22°C, 16:8 light:dark conditions and light intensity not exceeding 1000-1500 lux, respectively. The evaluation of the NOEC was determined based on the number of offspring’s produced per living parent Daphnia. Defined concentrations of the test chemical led to a certain percentage reduction of the parthenogenetic reproduction rate at the end of the 21 day study period. The living offspring was counted daily along with the renewal of the test medium. However, test vessels were inspected daily for the occurrence of juveniles and marked accordingly. On the basis of the effect on reproduction of the test daphnids, the 21 d NOEC and LOEC values was determined to be 0.175 and 0.228mg/l. Thus based on the outcomes chemical could be classified into aquatic chronic category 3 as per CLP classification criteria.

Key value for chemical safety assessment

Fresh water invertebrates

Effect concentration:: 0.175 mg/L

Additional information

Data for the long term toxicity of test chemical and structually and functionally similar read across chemicals on the mobility and mortality of test organism daphnia were summarized and studies are as mention below:

Based on the prediction, the long term toxicity on aquatic invertebrate daphnia species was predicted for test substance. On the basis of no effects observed in a freshwater daphnia, the NOEC value for the test substance is estimated to be 0.136 mg/l for daphnia for 21 days of exposure duration. Based on this value, it can be concluded that the test chemical was toxic to daphnia at environmentally relevant concentrations and chemical was readily biodegradable in water thus can be considered to be classified as aquatic chronic 3 category as per the CLP classification criteria.

Above toxic nature of test chemical was supported by the second study. Based on the prediction, the long term toxicity on aquatic invertebrate daphnia magna was predicted for test substance. On the basis of no effects observed in a freshwater daphnia, the NOEC value for the test substance is estimated to be 0.170 mg/l for daphnia for 21 days of exposure duration. Based on this value, it can be concluded that the test chemical can be considered as toxic to daphnia at environmentally relevant concentrations and chemical was readily biodegradable in water thus can be considered to be classified as aquatic chronic 3 category as per the CLP classification criteria.

Thus based on the above data it was concluded that the chemical was toxic and as it is readily biodegradable in water and thus chemical classified as aquatic chronic 3 category as per the CLP classification criteria.

Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.

Day	0	1	2	3	4	5	6	7	8	9	10	11	12	13	14	15	16	17	18	19	20	21
Medium renewal	-	-	Y	-	Y	-	Y	-	Y	-	-	Y	-	Y	-	Y	-	-	Y	-	Y	-
pH (new)	7.4	-	7.9	-	7.3	-	7.8	-	7.8	-	-	7.4	-	7.3	-	7.5	-	-	7.4	-	7.5	-
O2 (mg/l) (new)	7.5	-	8.0	-	7.8	-	7.9	-	7.8	-	-	7.8	-	7.9	-	7.6	-	-	7.9	-	7.7	-
Temp (°C) (new)	18	-	21	-	20.7	-	21.0	-	21.6	-	-	21. 8	-	21.3	-	21.5	-	-	21.9	-	21.5	-
Food provided	Y	-	Y	-	Y	-	Y	-	Y	-	-	Y	-	Y	-	Y	-	-	Y	-	Y	-
Replicates	Number of living offspring per replicate																						Total no. of live offsprings per vessel
V1	-	-	-	-	-	-	-	-	-	-	-	-	-	15	-	-	15	-	11	16	-	-	57
V2	-	-	-	-	-	-	-	-	-	-	-	-	10	-	-	26	-	-	9	-	-	-	45
V3	-	-	-	-	-	-	-	-	-	-	-	-	-	-	10	-	-	-	10	-	11	26	57
V4	-	-	-	-	-	-	-	-	-	-	-	25	-	15	16	4	-	-	7	-	13	-	80
V5	-	-	-	-	-	-	-	-	-	-	-	7	-	20	-	-	21	-	12	7	-	-	67
V6	-	-	-	-	-	-	-	-	-	-	-	15	-	25	-	-	-	-	16	-	14	-	70
V7	-	-	-	-	-	-	-	-	-	-	-	-	15	-	6	-	19	-	21	-	21	-	82
V8	-	-	-	-	-	-	-	-	-	-	-	-	-	11	-	-	15	-	12	15	-	-	53
V9	-	-	-	-	-	-	-	-	-	-	-	-	18	-	-	-	13	-	7	11	-	-	49
V10	-	-	-	-	-	-	-	-	-	-	-	-	10	-	5	-	-	-	21	-	-	16	52
Cumulative Reproductivity																							612

No. of Days		0	1	2	3	4	5	6	7	8	9	10	11	12	13	14	15	16	17	18	19	20	21
V1	Dead P	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V1	Dead O	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V2	Dead P	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V2	Dead O	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V3	Dead P	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V3	Dead O	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V4	Dead P	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V4	Dead O	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V5	Dead P	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V5	Dead O	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V6	Dead P	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V6	Dead O	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V7	Dead P	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V7	Dead O	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V8	Dead P	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V8	Dead O	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V9	Dead P	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V9	Dead O	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V10	Dead P	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V10	Dead O	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-

Day	0	1	2	3	4	5	6	7	8	9	10	11	12	13	14	15	16	17	18	19	20	21
Medium renewal	-	-	Y	-	Y	-	Y	-	Y	-	-	Y	-	Y	-	Y	-	-	Y	-	Y	-
pH (new)	8.0	-	8.0	-	7.8	-	7.6	-	8.1	-	-	7.3	-	7.5	-	7.4	-	-	7.5	-	7.6	-
O2 (mg/l) (new)	7.7	-	8.2	-	8.1	-	8.0	-	7.6	-	-	7.6	-	7.9	-	7.5	-	-	7.4	-	7.8	-
Temp (°C) (new)	18	-	21	-	20.7	-	21.0	-	21.8	-	-	21.8	-	21.3	-	21.5	-	-	21.9	-	21.5	-
Food provided	Y	-	Y	-	Y	-	Y	-	Y	-	-	Y	-	Y	-	Y	-	-	Y	-	Y	-
Replicates	Number of living offspring per replicate																						Total no. of live offspring’s per vessel
V1	-	-	-	-	-	-	-	-	-	-	-	-	-	8	-	-	17	-	-	-	-	-	25
V2	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	10	-	-	14	-	-	22	46
V3	-	-	-	-	-	-	-	-	-	-	-	-	6	-	-	-	-	-	10	17	-	-	33
V4	-	-	-	-	-	-	-	-	-	-	-	12	11	-	-	25	10	15	-	-	6	11	90
V5	-	-	-	-	-	-	-	-	-	-	-	-	-		9	-	5	-	7	11	11	-	43
V6	-	-	-	-	-	-	-	-	-	-	-	10	-	-	10	-	2	-	14	-	-	20	56
V7	-	-	-	-	-	-	-	-	-	-	-	-	-	-	8	-	7	-	-	17	21	-	53
V8	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	12	-	-	13	16	-	17	58
V9	-	-	-	-	-	-	-	-	-	-	-	12	-	-	15	-	4	-	-	13	-	9	53
V10	-	-	-	-	-	-	-	-	-	-	-	1	-	15	-	-	13	-	6	8	12	-	55
Cumulative Reproductivity																							512

No. of Days		0	1	2	3	4	5	6	7	8	9	10	11	12	13	14	15	16	17	18	19	20	21
V1	Dead P	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V1	Dead O	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V2	Dead P	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V2	Dead O	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V3	Dead P	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V3	Dead O	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V4	Dead P	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V4	Dead O	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V5	Dead P	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V5	Dead O	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V6	Dead P	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V6	Dead O	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V7	Dead P	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V7	Dead O	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V8	Dead P	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V8	Dead O	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V9	Dead P	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V9	Dead O	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V10	Dead P	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V10	Dead O	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-

Day	0	1	2	3	4	5	6	7	8	9	10	11	12	13	14	15	16	17	18	19	20	21
Medium renewal	-	-	Y	-	Y	-	Y	-	Y	-	-	Y	-	Y	-	Y	-	-	Y	-	Y	-
pH (new)	8.1	-	7.7	-	8.0	-	7.7	-	8.1	-	-	7.5	-	7.5	-	7.6	-	-	7.5	-	7.6	-
O2 (mg/l) (new)	7.7	-	8.0	-	8.2	-	7.9	-	7.8	-	-	7.6	-	7.8	-	7.6	-	-	7.3	-	7.7	-
Temp (°C) (new)	18	-	21	-	20.7	-	21.0	-	21.8	-	-	21.8	-	21.3	-	21.5	-	-	21.9	-	21.5	-
Food provided	Y	-	Y	-	Y	-	Y	-	Y	-	-	Y	-	Y	-	Y	-	-	Y	-	Y	-
Replicates	Number of living offspring per replicate																						Total no. of live offspring’s per vessel
V1	-	-	-	-	-	-	-	-	-	-	-	7	-	-	17	-	-	-	14	-	22	-	60
V2	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	11	-	-	9	-	-	-	20
V3	-	-	-	-	-	-	-	-	-	-	-	-	-	-	11	-	-	-	6	-	-	-	17
V4	-	-	-	-	-	-	-	-	-	-	-	-	-	10	-	-	7	-	-	10	-	-	27
V5	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	7	-	-	5	-	-	5	17
V6	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	15	-	-	9	-	-	-	24
V7	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	12	21	-	1	-	21	-	55
V8	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	3	-	-	1	-	5	-	9
V9	-	-	-	-	-	-	-	-	-	-	-	-	-	-	9	-	-	-	8	1	17	-	35
V10	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	1	13	-	-	7	-	-	21
Cumulative Reproductivity																							285