6-chloro-5-(2-chloroethyl)-1,3-dihydroindol-2-one

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:

18 April - 16 May 1996

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1996

Report date:

1996

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

other: EC Directive 92/69/EEC, Method C4-E

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 301 D (Ready Biodegradability: Closed Bottle Test)

Principles of method if other than guideline:

Sealed bottles containing the test substance and inorganic mineral medium were inoculated with activated sewage sludge bacteria and incubated for up to 28 days at 20 ± 1°C. On Days 0, 4, 7, 11, 14, 18, 21, 25 and 28 duplicate bottles were removed and dissolved oxygen measurements were performed electrochemically. Percentage biodegradation values were calculated by comparing the
extent of oxygen depletion with the Theoretical Oxygen Demand (ThODN03 = 1.74 mg02/mg). Additional bottles, containing both the test substance and a readily biodegradable standard substance were prepared in order to provide additional information on the inhibitory effect of the test substance (Days 0, 14 and 28 only

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

6-chloro-5-(2-chloroethyl)-1,3-dihydroindol-2-one

EC Number:

421-320-0

EC Name:

6-chloro-5-(2-chloroethyl)-1,3-dihydroindol-2-one

Cas Number:

118289-55-7

Molecular formula:

Hill formula: C10 H9 Cl2 N O CAS formula: C10 H9 Cl2 N O

IUPAC Name:

6-chloro-5-(2-chloroethyl)-2,3-dihydro-1H-indol-2-one

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

Use: Pharmaceutical intermediate
Date recieved: 2 February 1996
Purity: 99.9%

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
STANDARD SUBSTANCE
Sodium benzoate (C6H5.COONa).
Fisons A.R. Grade, Batch No. 9401096045, Purity >99.5%.

Study design

Oxygen conditions:

anaerobic

Inoculum or test system:

activated sludge, adapted

Details on inoculum:

Type: Activated sewage sludge bacteria.

Source: The aeration stage of the Anglian Water sewage treatment plant (Godmanchester) treating predominantly domestic sewage

Preparation: A sample was collected on 17 April 1996 and maintained under aeration for < 24 hours ,allowed to settle for 30 minutes and the supernatant filtered through Whatman GFA coarse filter paper (first 250 ml discarded). The filtrate was maintained under aeration until used.

Duration of test (contact time):

28 d

Initial test substance concentration

Initial conc.:

2 mg/L

Parameter followed for biodegradation estimation

Parameter followed for biodegradation estimation:

O2 consumption

Remarks:

Dissolved oxygen

Details on study design:

Biochemical Oxygen Demand bottles (280 ml) of darkened glass and fitted with ground glass stoppers were filled by siphon and firmly stoppered to exclude all air bubbles. Sufficient bottles were prepared in each test and control series to allow two bottles to be removed at each time point for a single oxygen determination per bottle to be made at 0, 4, 7, 11, 14, 18, 21, 25 and 28 days. Sufficient bottles for series (d) were prepared to provide information on inhibitory effects on Days 0, 14 and 28 only.

The bottles were incubated in a water bath at 20 ± 1°C until removed for measurement of dissolved oxygen concentration by means of a Yellow Springs BOD Meter

Reference substance

Reference substance:

other: Sodium benzoate

Results and discussion

Preliminary study:

N/A

Test performance:

Cultures containing both test and standard substances combined showed an oxygen depletion value equivalent to 40 % biodegradation based on the combined theoretical oxygen demands of the test and standard substances (1.74 and 1.67 mgO/mg respectively). Consequently CP-89,575 was not considered to have had an inhibitory effect on sewage bacteria under the conditions of this test.

Sodium benzoate attained 99% biodegradation within 28 days.

Oxygen depletions in the inoculated control series were within the prescribed limits ( 1.5 mgO/l after 28 days).

% Degradation

Key result

Parameter:

% degradation (DOC removal)

Value:

20

Sampling time:

28 d

Details on results:

Points of degradation plot (test substance):
5 % degradation after 4 d
7 % degradation after 7 d
5 % degradation after 11 d
6 % degradation after 14 d
6 % degradation after 18 d
12 % degradation after 21 d
0 % degradation after 25 d
20 % degradation after 28 d

BOD5 / COD results

Results with reference substance:

Points of degradation plot (reference substance):
90 % degradation after 4 d
80 % degradation after 7 d
82 % degradation after 11 d
78 % degradation after 14 d
97 % degradation after 18 d
99 % degradation after 21 d
91 % degradation after 25 d
99 % degradation after 28 d

Any other information on results incl. tables

Culturemedium		Day
		0	4	7	11	14	18	21	25	28
(a)       Mineralmediumwithinnculum       (b)      Testsubstance(2mg/I)       (c) Standardsubst3ncc(3m/l)Sodiumbenzoatc --   (d)Testsubsta1K'C{21110:/IJplusstandardsubstance(3 m/l)	R1R2 Mean   R1 R2 Mean   R1 R,       RIR, Mean	9.030 9.030 9.030   9.030 9.030 9.030   9.030 9.030 9.030   9.030 9.030 9.030	9.010 8.820 8.915   8.630 8.820 8.725   4.820 3.970 4.395	8.540 8.650 8.595   8.320 8.400 8.360   4.62 4.53 4.575	8.340 8.210 8.275   8.260 7.970 8.115   4.280 4.090 4.185	8.330 7.910 8.120   7.910 7.890 7.900   4.340 4.100 4.2'.W   4.500 5.000 4.750	7.770 7.950 7.860   7.540 7.770 7.655   3.050 2.980 3.015	7.830 8.010 7.920   7.440 7.570 7.505   3.070 2.880 2.975	7.680 7.300 7.490   7.540 7.510 7.525   2.900 2.960 2.930	7.700 7.750 7.725   7.090 6.960 7.025   2.710 2.860 2.785   3.840 3.600 3.720

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Interpretation of results:

other: not readily biodegradable

Conclusions:

CP-89,575 may not be termed as readily biodegradable.

There was no evidence of inhibitory effects on the activity of the activated sewage sludge under the conditions of this test.