cis-4-[3-(p-tert-butylphenyl)-2-methylpropyl]-2,6-dimethylmorpholine

Bioaccumulation: aquatic / sediment

Administrative data

Endpoint:

bioaccumulation in aquatic species: fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

18.12.2001- 19.09.2002

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2003

Report date:

2003

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 305 (Bioconcentration: Flow-through Fish Test)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.1730 (Fish Bioconcentration Test)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

cis-4-[3-(p-tert-butylphenyl)-2-methylpropyl]-2,6-dimethylmorpholine

EC Number:

266-719-9

EC Name:

cis-4-[3-(p-tert-butylphenyl)-2-methylpropyl]-2,6-dimethylmorpholine

Cas Number:

67564-91-4

Molecular formula:

C20H33NO

IUPAC Name:

(2R,6S)-4-[3-(4-tert-butylphenyl)-2-methylpropyl]-2,6-dimethylmorpholine

Details on test material:

other TS: 14-C Fenpropimorph, morpholine-labelled

Radiolabelling:

yes

Remarks:

Phenyl- and Morpholine- labelled: PhenyI-U-14C-Label: Lot Number: 471-2201; Chemical Purity: >96%; Radiochemical Purity: >96% Morpholine-2(6)—Label: Lot Number: 639-2101; Chemical Purity: >96%; Radiochemical Purity: >98%

Sampling and analysis

Details on sampling:

Samgling of Water:
Two aliquots of 5ml were taken from each tank on the last two days of the pre-equilibration period (Day —1 and Day 0), on Days 1, 3, 7, 14, 17, 19, and 20 of the uptake period and on Days 1, 2, 3, 4, 8, and 16 of the depuration period. The aliquots were directly analysed for total radioactivity at the Experimental Toxicology and Ecology Department (GV/TC). Further aliquots of 4 x 1 L and 1 x 0.5 L of tank water were taken from each dose group R6 and R7 on Days 14 and 20 for quantification and identification of metabolites in water at the
BASF Agricultural Center, Limburgerhof.

Samplinq of Fish Tissue:
Five fish were removed from the main tanks R6 and R7 on days 1, 3, 7, 14, 17 and 19 of the uptake period and on days 1, 4, 8, 16, 23 and 56 of the depuration period. The fish were dissected into filet as edible portion of the fish, and into head, fins and skin, and the gastrointestinal tract as inedible portions as preparation for analysis of total radioactivity.
In addition to the sampling of the dose groups R6 and R7, the same number of fish was removed from the control group R1 at each sampling time. The fish of the control group were further split into a sample of two fish for blank correction and into a sample of three fish for lipid determination.

Test organisms

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Common name: Rainbow trout
- Source: Test groups (R1, R6. R7*): Fish from the same batch with same origin and same hatch date. Supplier: Forellenzucht Hoch-Spessart, 97840 Erlenfurt, Germany
Satellite groups (R6a. R7a*): Fish from the same batch with same origin and same hatch date. Supplier: Forellenzucht Trostadt, 98646 Trostadt, Germany
- Age at study initiation (mean and range, SD):
Hatch dates: Test groups (R1, R6, R7): 5- 7 Jan, 2002
Satellite groups (R6a. R7a): 10 Dec, 2001
- Length at study initiation (length definition, mean, range and SD):
Test groups (R1, R6. R7): 4.7 —5.6 cm
Satellite groups (R6a. R7a): 6.5 - 8.7 cm
- Weight at study initiation (mean and range, SD):
Test groups (R1, R6, R7): 0.9 - 1.4 g. the minimum body weight was
about 2/3 of the maximum body weight.
Satellite groups (R6a, R7a): 3.6 —7.5 9
- Health status: Healthy
- Feeding during test
- Food type: Growing feed ,,Foreltenfutter (Zeigler)", supplier Provimi Kliba AG, Gossau. Switzerland, ad libitum, additionally generally on workdays live or frozen brine shrimp (artemia). The composition is suitable for the test species.
- Amount: During test and adaptation approx. 1 - 2 % of the mean body weight per day; generally in 2 applications per day, on saturdays, sundays and holidays 1 application per day.


ACCLIMATION
- Acclimation period: at least 14 days
- Acclimation conditions: same
- Type and amount of food: During test and adaptation approx. 1 - 2 % of the mean body weight per day; Growing feed ,,Foreltenfutter (Zeigler)", supplier Provimi Kliba AG, Gossau. Switzerland, ad libitum, additionally generally on workdays live or frozen brine shrimp (artemia). The composition is suitable for the test species.
- Feeding frequency:
- Health during acclimation (any mortality observed): The test fishes were not treated. Experience of the testing laboratory with carp purchased from the supplier shows that generally the fish are in a stable health condition at
the time of delivery. The animals were visually inspected for their state of health before begin of adaptation and before study start. Only healthy fish were used. The mortality during the adaptation time was < 5% per week.

Study design

Route of exposure:

aqueous

Test type:

flow-through

Water / sediment media type:

natural water: freshwater

Total exposure / uptake duration:

20 d

Total depuration duration:

56 d

Test conditions

Hardness:

2.5 - 3.8 mmol CaCO3/L, corresponds to 250 - 380 mg CaCO3/L.

Test temperature:

15 ± 1°C

pH:

7.8 - 8.2.

Dissolved oxygen:

> 60%

TOC:

1.9 - 3.6 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Glas aquaria with silicon rubber seals, provided with an overflow. Dimensions: 80 cm length x 35 cm width x 55 cm height, water volume 100 L. 1 test vessel was used per test group.
- Aeration: Since an oxygen concentration of > 60% in the test vessel was not ensured. the test vessels were slightly aerated. The aeration had no effect on the concentration of the test substance in the test vessels.
- Renewal rate of test solution (frequency/flow rate): Approx. 20 L/hour per test aquarium, this amounts to an approx. 5fold change of water per day in the test groups R1, R6 and R7.
The satellite groups R6a and R7a received the water from the outlet of the test vessels R6 and R7. respectively and had therefore a 12fold water exchange per day.
- No. of organisms per vessel: 100 in control and test; 16 in satallite groups
- No. of vessels per concentration (replicates): 2
- No. of vessels per control / vehicle control (replicates): 1
- Biomass loading rate: At study start 100 fish with a mean body weight of 1.1 g (wet weight) were inserted into each test vessel of the test groups R1, R6 and R7. Therefore the loading was 0.22 g fish/L test water per day based on the ow rate and 1.1 g fish/L based on the water volume in the test vessel. During the exposure the loading rate was decreased rapidely by removing fish from the test vessels for sampling.
The mean body weight of the fish in the satellite groups was 4.8 g. The loading rate was therefore 0.15 g fish/L test water per day based on the ow rate and 1.9 g fish/L based on the water volume in the test vessel.

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Charcoal filtered tap water of the city of Frankenthal
- Intervals of water quality measurement: The tap water is regularty analysed for impurities by the city of Frankenthat as well as by BASF Aktiengesellschaft,
Department of Water Chemistry and Technicat Services. Regarding the objective and the duration of the study no specic requirements were dened for the dilution water which exceed the standard specication of drinking water.


OTHER TEST CONDITIONS
- Adjustment of pH: twice weekly
- Photoperiod: 16 hours light, 8 hours darkness

Nominal and measured concentrations:

nominal concentration: 0.2 µg/L;
measured concentration: test group R6 (Phenyl Label): 0.20 µg/L, R7 (Morpholin Label): 0.22 µg/L

Results and discussion

Bioaccumulation factor

Conc. / dose:

0.2 µg/L

Temp.:

> 14 - < 16 °C

pH:

8

Type:

BCF

Value:

> 968 - < 1 145 dimensionless

Basis:

whole body w.w.

Time of plateau:

19 d

Depuration

Depuration 1

Elimination:

yes

Parameter:

DT90

Depuration time (DT):

4.425 d

Remarks on result:

other: average of depuration of phenyl- and morpheoline-label

Depuration 2

Elimination:

yes

Parameter:

DT50

Depuration time (DT):

1.34 d

Remarks on result:

other: average of depuration of phenyl- and morpheoline-label

Metabolites:

BF421-1 (Reg. No. 246 719) and fenpropimorph-acid (BF421-2, Reg. No. 231 346)

Any other information on results incl. tables

Parameter	Tissue Fraction
	Phenyl-Label			Morpholine-Label
	Edibles	In-Edibles	Whole Fish	Edibles	In-Edibles	Whole Fish
Uptake rate constant k1 [days-1]	142.81	771.91	576.12	147.45	846.87	671.47
Depuration rate constant k2 [days-1]	0.46	0.50	0.49	0.34	0.61	0.55
Depuration half-life DT50 [days]	1.52	1.38	1.41	2.05	1.14	1.26
Time to reach 90% depuration DT90 [days]	5.03	4.57	4.67	6.79	3.79	4.18
Bioconcentration factor (BCFk) k1/k2	312	1533	1169	435	1394	1220

    
     

Applicant's summary and conclusion

Conclusions:

The uptake of fenpropimorph in rainbow trout showed to have reached steady-state conditions within 19 days. Calculation of the bioconcentration factor BCFss based on measured radioactivity in tank water and fish resulted in 1145 for whole fish of the Phenyl Label and correspondingly in 968 for the Morpholine Label at steady-state. The depuration of 95% of the residues at the plateau level lasted up to about 56 days. A non-linear regression analysis showed that the data could be adequately described using a one-fish-compartment model, although the depuration rate was slightly overestimated. One of the kinetic parameters determined was the depuration half-life time with 1.4 days for the Phenyl Label and with 1.3 days for the Morpholine Label. The time of 90% elimination was calculated to 4.7 days and 4.2 days, respectively. The kinetic bioconcentration factors BCFk as the ratio of the uptake rate and the depuration rate constants resulted in 1169 and 1220 for whole fish of the Phenyl and of the Morpholine Label, respectively. These data are very similar to the BCFss values from direct calculation of the radioactivity measurements of water and fish samples at steady-state.

Metabolisation of fenpropimorph was detected in edible and inedible fish tissue accordingly.
Predominant metabolite was the glucuronic acid conjugate BF 421-1-Gluc.A. with up to more than 50% of the radioactive residue. This metabolite was also detected in the tank water at small amounts besides mainly the parent compound.