Anthraquinone acid dye

Biodegradation in water: screening tests

Administrative data

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

key study

Study period:

16 April 2009 to 14 May 2009

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted to GLP in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2009

Report date:

2009

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

other: "Method for Testing the Biodegradability of Chemical Substances by Microorganisms" stipulated in the "Testing Methods for New Chemical Substances" (No. 1121002, Pharmaceutical and Food Safety Bureau, Ministry of Health, Labour and Welfare...

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 301 C (Ready Biodegradability: Modified MITI Test (I))

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Details on test material:

- Name of test material (as cited in study report): Ink BH11 M

Study design

Oxygen conditions:

not specified

Inoculum or test system:

activated sludge (adaptation not specified)

Details on inoculum:

Activated sludge was collected, prepared and controlled by Chemicals Evaluation and Research Institute according to the test methods described. Activated sludge was cultivated for 18.5 hours after the synthetic sewage was added. Synthetic sewage was prepared with glucose, peptone and potassium dihydrogenphosphate were dissolved in purified water, and the pH of the solution was adjusted to 7.0 +/- 1.0.

Duration of test (contact time):

28 d

Initial test substance concentration

Initial conc.:

100 mg/L

Based on:

formulation

Parameter followed for biodegradation estimation

Parameter followed for biodegradation estimation 1

Parameter followed for biodegradation estimation:

DOC removal

Parameter followed for biodegradation estimation 2

Parameter followed for biodegradation estimation:

other: Biological Oxygen Demand (BOD)

Details on study design:

TEST CONDITIONS
- Composition of medium: Not specified
- Additional substrate: Each of the test solutions were made up to 1000 mL with purified water.
- Solubilising agent (type and concentration if used): n/a
- Test temperature: 25±1°C
- pH: 7.0
- pH adjusted: Yes
- Aeration of dilution water: Not specified
- Suspended solids concentration: The activated sludge was added to each test vessel so that the concentration of the suspended solid
reached 30 mg/L.
- Continuous darkness: Yes
TEST SYSTEM
- Culturing apparatus: Closed system oxygen consumption measuring apparatus
- Number of culture flasks/concentration: 3
- Method used to create aerobic conditions: Not specified
- Method used to create anaerobic conditions: n/a
- Measuring equipment: Closed system oxygen consumption measuring apparatus
(Temperature controlled bath and measuring unit : Asahi Techneion Co., Ltd.)
(Data sampler : Asahi Techneion Co., Ltd.)
Vessel 300 mL in volume (Improved type)
Absorbent for carbon dioxide - Soda lime No.1 (for absorption of carbon dioxide, Wako Pure Chemical Industries, Ltd.)
- Test performed in closed vessels due to significant volatility of test substance: No
- Test performed in open system: Not specified
SAMPLING
- Sampling frequency: Not specified
- Sampling method: Not specified
- Sample storage before analysis: Not specified
CONTROL AND BLANK SYSTEM
- Inoculum blank: In one test vessel, nothing was added to the basal culture medium


STATISTICAL METHODS:
-The concentration of DOC (Dissolved Organic Carbon) in the sample for TOC was calculated by subtracting concentration of the inorganic
carbon (IC) from concentration of the total carbon (TC). The concentration of TC and IC in the test solutions was proportionally calculated
from the peak area of the test solution by comparison with that of 80.0 mgC/L standard solution for TC analysis and 40.0 mgC/L standard
solution for IC analysis, respectively.

The standard solution for TC analysis was prepared by dissolving potassium hydrogen phthalate in purified water. The standard solution for IC analysis was prepared by dissolving sodium hydrogen carbonate and sodium carbonate in purified water.

The test item was detected as four peaks on the chromatogram. Then, the concentration of the test item in the sample for HPLC analysis was calculated proportionally by comparing the total peak area on the chromatogram of the sample for HPLC analysis with that on the chromatogram of 101 mg/L standard solution
The lowest detectable peak area of the test item was regarded as 3000 considering the noise level, which corresponded to the test item concentration of 0.17 mg/L.

Percentage biodegradation by BOD:
Percentage biodegradation (%) = (BOD - B/TOD) x 100
BOD = Biochemical oxygen demand in the test solution
B = Biochemical oxygen demand in the control blank
TOD = Theoretical oxygen demand required when the test item was completely oxidized


Percentage biodegradation by DOC:
Percentage biodegradation (%) = (DOCw - DOCs/DOCw) x 100

DOCs = Residual amount of the dissolved organic carbon in the test solution
DOCw = Residual amount of the dissolved organic carbon in the test solution


Percentage biodegradation of test item:
Percentage biodegradation (%) = (Sw - Ss/Sw) x 100
Ss = Residual amount of the test item in the test solution
Sw = Residual amount of the test item in the test solution

Reference substance

Reference substance:

aniline

Results and discussion

% Degradation

Parameter:

other: Biological Oxygen Demand (BOD)

Value:

0

Details on results:

Results
The average percentage biodegradations by BOD and of test item were regarded as 0% because the calculated average values were negative.

BOD5 / COD results

Results with reference substance:

Day 7 70% degradability
Day 14 77% degradability
Day 21 78% degradability
Day 28 77% degradability

Any other information on results incl. tables

Results

The average percentage biodegradations by BOD and of test item were regarded as 0% because the calculated average values were negative.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Interpretation of results:

not inherently biodegradable

Conclusions:

The test item was not biodegraded by microorganisms under the test conditions of this study.

Executive summary:

In a biodegradation study of Ink BH11 M by microorganisms (CERI study number: 15275) the test material was

considered not biodegraded. The method followed that described in the OECD Guidelines for Testing of Chemicals, Guideline 301 C, Revised July 17, 1992, Ready Biodegradability: Modified MITI Test (I)