Phosphoric acid, hexadecyl esters, potassium salts with cetyl alcohol and isostearyl isostearate

Toxicity to microorganisms

Administrative data

Endpoint:

activated sludge respiration inhibition testing

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

the study does not need to be conducted because the substance is found to be readily biodegradable and the applied test concentrations are in the range of concentrations that can be expected in the influent of a sewage treatment plant

Cross-reference

Reason / purpose for cross-reference:

data waiving: supporting information

Reference

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From October 24, 2017 to November 24, 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 310 (Ready Biodegradability - CO2 in Sealed Vessels (Headspace Test)

Deviations:

not specified

GLP compliance:

yes

Oxygen conditions:

aerobic

Inoculum or test system:

other: Aerobic sludge, mixed treatment plant of urban (66%) and industrial (34%)

Details on inoculum:

Method of mud sampling
For the preparation of the inoculum a sample of aerobic sludge has been selected by the mixed treatment plant of urban (about 66%) and industrial (about 34%) liquid sewage situated at San Rocco - Monza (MB), Italia. The plant of treatment is managed by "BRIANZACQUE SRL (Monza)".

Mud preparation
In the laboratory the sampled muds, have been mixed and let settle, keeping them in aerobic conditions. The mud samples, before their use, have been analysed to check its ability to form colony units. Then it has been centrifuged, washed and analyzed to quantify the suspended solids concentration for the inoculum preparation. The inoculum was prepared in order to have a concentration of suspended solid of about 4 mg/L.

Medium preparation:

Solution A
Anhydrous potassium dihydrogen phosphate (KH2P04): 8.50 g/L
Anhydrous dipotassium hydrogen phosphate (K2HP04): 21.75 g/L
Disodium hydrogen phosphate dihydrate (Na2HP04.2H20): 33.40 g/L
Ammonium chloride (NH4Cl): 0.50 g/L
pH = 7.53

Solution B
Calcium chloride dihydrate (CaCI2.2H20): 36.40 g/L

Solution C
Magnesium sulphate heptahydrate (MgSO4.7H20): 22.50 g/L

Solution D
Iron chloride (Ill) hexahydrate (FeCl3.6H20): 0.25 g/L
All solutions has been made with deionized water (MilliQ Millipore). Then for each litre of medium 10 mL of Solution A have been transferred into a 1000 mL volumetric flask and diluted with 800 mL of deionised water; 1 mL of Solution B, 1 mL of Solution C and 1 mL of Solution D have been added and the solution has been diluted to volume with deionised water. For higher volumes the medium has been prepared according to the same proportion. The preparation of the solutions has been reported in an internal laboratory logbook.

Duration of test (contact time):

ca. 28 d

Initial conc.:

ca. 39.73 mg/L

Based on:

TOC

Parameter followed for biodegradation estimation:

inorg. C analysis

Reference substance:

benzoic acid, sodium salt

Remarks:

18.61 mg/L of TOC

Key result

Parameter:

% degradation (inorg. C analysis)

Remarks:

Test substance

Value:

ca. 118

Sampling time:

28 d

Remarks on result:

other: Readily biodegradable

Results with reference substance:

Percentage biodegradation of reference substance at 14 d was determined to be 101%, therefore the assay fulfilled the validity criteria.

Results

Quality criteria of the test were satisfied. The mean amount of TIC present in the blank controls at the end of the test was < 3 mg C/L. The trend of the inorganic carbon and related biodegradation percentages in the bottle contained both the test sample and the reference substance confirm the absence of inhibitory effect of the test sample on inoculum. The biodegradation percentage of the test substance during the test was as follow:

Check point (d)	% of Biodegradation Reference substance	% of Biodegradation Test substance	% of Biodegradation Reference + Test substance
0	0	0	0
1	3	3	7
7	90	35	-
14	101	58	-
21	102	88	-
28	127	118	90

 

Conclusions

On the basis of results obtained, interpreted in accordance to OECD 310:2014, the test substance was considered readily biodegradable in aerobic condition.

Validity criteria fulfilled:

yes

Interpretation of results:

readily biodegradable

Conclusions:

Under the study conditions, the test substance was determined to be readily biodegradable.

Executive summary:

A study was conducted to determine the ready biodegradability of the test substance, 'mono- and di- C16 PSE, K+ and C16-OH and isostearyl isostearate', using Headspace test method (CO2 in sealed vessels), according to OECD Guideline 310, in compliance with GLP. Aerobic sludge inoculum was prepared with mixed treatment plant of urban (66%) and industrial (34%) liquid sewage and in order to have a concentration of suspended solid of about 4 mg/L. In the study, 17 replicates of blanks (containing only culture medium), positive control (sodium benzoate at 18.61 mgTOC/L) and test substance (39.73 mg TOC/L) and 8 replicates of toxicity control (i.e., Test and reference substances (49.77 mg TOC/L)) were used. All treated samples were kept at the temperature of 20°C for 28 d. The calculation of biodegradation as total inorganic carbon (TIC) was determined at each sampling time (i.e., 0, 1, 7, 14, 21 and 28 d) for the reference substance, test sample and blank. For toxicity control, calculation of biodegradability as total inorganic carbon (TIC) was determined on Day 1 and 28. The trend of the inorganic carbon and related biodegradation percentages in the bottle containing both the test sample and the reference substance, confirmed the absence of inhibitory effect of the test sample on inoculum. At 28 d, the biodegradability of the test substance was determined to be 118%. At 14 d biodegradability for reference substance was determined to be 101%. The mean amount of TIC present in the blank controls at the end of the test was < 3 mg C/L. Based on the study results, the test had satisfied all the validity criteria. Therefore, under the study conditions, the test substance was considered to be readily biodegradable (Eurofins, 2017).