Disodium 4,4'-bis[(4-anilino-6-morpholino-1,3,5-triazin-2-yl)amino]stilbene-2,2'-disulphonate

Endpoint summary

Description of key information

Bioaccumulation: aquatic / sediment

Bioaccumulation study was conducted on test organism Cyprinus carpiofor evaluating the bioconcentration factor (BCF value) of test chemical (Authoritative database, 2019 and secondary sources). The study was performed in accordance with the OECD TG 305 C "Bioaccumulation: Test for the Degree of Bioconcentration in Fish" and other guideline "Bioaccumulation test of a chemical substance in fish or shellfish" provided in "the Notice on the Test Method Concerning New Chemical Substances", respectively at 25±2°C and dissolved oxygen content of 6 -8 mg/l for 6 weeks. Cyprinus carpio (weight ca. 30 gm, length ca. 10 cm) supplied by Sugishama fish farm was used as a test organism for the study. Lipid content of the test organism at the start of exposure was 5.2%. Test organisms were fed with pelleted food about 1 % of body weight twice/day. Fishes were reared in an acclimatization tank (flow through system) for 28 days. Details on analytical methods involve the recovery ratio: Test water: 1st concentration area: 94.6%, 2nd concentration area : 91.3%, Fish: 85.9%, - Limit of quantitation: Fish : 0.093 ppm. Test chemical nominal conc. used for the study was 0.2 mg/land 0.02 mg/l. Dimethyl sulfoxide was used as a vehicle. Range finding study involves the TLm (48 hr) ≥ 50 ppm (w/v) on Rice fish (Oryzias latipes). Glass tank was used as a test vessel. Test vessel contains 15 -20 fish per level in glass tank. Total hardness, COD, chloride, and other parameters were measured every 6 months. In addition to this, water temperature, pH and dissolved oxygen were also measured continuously. The bioconcentration factor (BCF value) of test chemical on Cyprinus carpio was determined to be in the range of 1.4– 4.7 L/Kg at a conc. of 0.2 mg/l and 6.4 – 28 L/Kg at a conc. of 0.02 mg/l, respectively, which does not exceed the bioconcentration threshold of 2000, indicating that the test chemical is not expected to bioaccumulate in the food chain.

Additional information

Bioaccumulation: aquatic / sediment

Various experimental studies of the test chemical were reviewed for the bioaccumulation end point which are summarized as below:

 

In an experimental key study from authoritative database (2019) and secondary sources, bioaccumulation study was conducted on test organism Cyprinus carpio for evaluating the bioconcentration factor (BCF value) of test chemical. The study was performed in accordance with the OECD TG 305 C "Bioaccumulation: Test for the Degree of Bioconcentration in Fish" and other guideline "Bioaccumulation test of a chemical substance in fish or shellfish" provided in "the Notice on the Test Method Concerning New Chemical Substances", respectively at 25±2°C and dissolved oxygen content of 6 -8 mg/l for 6 weeks. Cyprinus carpio (weight ca. 30 gm, length ca. 10 cm) supplied by Sugishama fish farm was used as a test organism for the study. Lipid content of the test organism at the start of exposure was 5.2%. Test organisms were fed with pelleted food about 1 % of body weight twice/day. Fishes were reared in an acclimatization tank (flow through system) for 28 days. Details on analytical methods involve the recovery ratio: Test water: 1st concentration area: 94.6%, 2nd concentration area : 91.3%, Fish: 85.9%, - Limit of quantitation: Fish : 0.093 ppm. Test chemical nominal conc. used for the study was 0.2 mg/land 0.02 mg/l. Dimethyl sulfoxide was used as a vehicle. Range finding study involves the TLm (48 hr) ≥ 50 ppm (w/v) on Rice fish (Oryzias latipes). Glass tank was used as a test vessel. Test vessel contains 15 -20 fish per level in glass tank. Total hardness, COD, chloride, and other parameters were measured every 6 months. In addition to this, water temperature, pH and dissolved oxygen were also measured continuously. The bioconcentration factor (BCF value) of test chemical on Cyprinus carpio was determined to be in the range of 1.4– 4.7 L/Kg at a conc. of 0.2 mg/l and 6.4 – 28 L/Kg at a conc. of 0.02 mg/l, respectively.

 

Another bioaccumulation study was conducted on test organism Lepomis macrochirus (Bluegill) for evaluating the bioconcentration factor (BCF value) of test chemical (OECD SIDS, 2005). The study was performed under flow through conditions with continuous aeration (flow rate of 6 l/hour) at 18 ± 0.5°C, pH 7.3, dissolved oxygen content of ≥ 50 mg/l and hardness of 40 mg/l CaCo3 for 70 days. Lepomis macrochirus(Bluegill) (weight ca. 50 gm, length ca. 140 mm) was used as a test organism for the study. Test chemical conc. used for the study were0, 0.001, 0.01, 0.1 and 1 mg/l, respectively.Analysis of the water samples showed that the concentration of test chemical in the test vessel was usually within ±15 % of the nominal levels. In the 1 mg/l test vessel of the repeated study, the recovery was between 80 to 110 %, depending on the sampling day. Test chemical was not found above the quantizable limit in fish samples taken on days 28, 42 and 70 during the accumulation period and after the withdrawal of samples on day 1, 3, 7 and 14, respectively.Tank was used as a test vessel. 60 fishes/ vessel were exposed to the various test chemical conc. Fish were sampled on days 1, 3, 7, 14, 21, 28, 56 and 70 of exposure. After the termination of the study, fishes were placed in uncontaminated water and were sampled on days 1, 3, 7 and 14. At each sampling day, 3 fish were samples from each tank. Viscera and carcasses were analyzed for test chemical. The bioconcentration factor (BCF value) of test chemical on Lepomis macrochirus (Bluegill) was determined to be < 1 at a conc. of 1 mg/l.

 

In a supporting study from handbook (2008) and secondary source (2004), bioaccumulation experiment was conducted on test organism Leuciscus idus for evaluating the bioconcentration factor (BCF value) of test chemical. Test fishes were exposed to nominal test chemical conc. of 0.00005 mg/l (50 ppb) (dynamic system) and 0.000005 mg/l (5 ppb) (static system). After 1, 3 and 7 days, exposed three fish were sampled each time and radioactivity was measured of different parts. The bioconcentration factor (BCF value) of test chemical on Leuciscus idus in the filet and the total fish was determined to be < 1 at a conc. of 0.000005 to 0.00005 mg/l.

 

In an additional study from secondary source (2005), bioaccumulation study was conducted on test organism Leuciscus idus for evaluating the bioconcentration factor (BCF value) of test chemical. Test chemical nominal conc. used for the study 0.1 mg/l (100 µg/l) (dynamic system) and 0.01 mg/l (10 µg/l) (static system). Precipitation of the test substance was observed after 2-3 days in the stock solution. After 1, 3 and 7 days fish (n = 3) were sampled and different parts of the fish bodies like viscera, gills, head, skin, fillet were analyzed for test chemical. Study was performed in duplicates and mean BCF values were calculated. The bioconcentration factor (BCF value) of test chemical on Leuciscus idus was determined to be in the range of 0.5 to 2.4 L/Kg at a conc. of 0.1 mg/l.

 

On the basis of above results for test chemical(from handbook, authoritative databases and secondary sources), it can be concluded that the BCF value of test chemicalwas determined to be ranges from 0.5 to 28, respectively,which does not exceed the bioconcentration threshold of 2000, indicating that the test chemical is not expected to bioaccumulate in the food chain.

Bioaccumulation endpoint can also be considered for waiver as per inaccordance with column 2 of Annex IX of the REACH regulation, testing for this endpoint is scientifically not necessary and does not need to be conducted since the test chemical has a low potential for bioaccumulation based on logKow ≤ 3.