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Environmental fate & pathways

Bioaccumulation: aquatic / sediment

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Administrative data

Endpoint:
bioaccumulation in aquatic species: fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
25/July/2002 to 22/August/2002
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The work reported was performed carefully, the results reasonable, and the conclusions conservative. Data was generated according to generally valid and internationally accepted testing guidelines.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2002
Report Date:
2002

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
other:
Deviations:
no
Principles of method if other than guideline:
“Method for Testing the Degree of Accumulation of Chemical Substances in Fish Body” in “Testing Methods for New Chemical Substances” (July 13, 1974, Revised October 8, 1998, No.5, Planning and Coordination Bureau, Environment Agency; No.615, Pharmaceutical Affairs Bureau, Ministry and Health and Welfare; and No.392, Basic Industries Bureau, Ministry of International Trade and Industry, Japan), and “Bioconcentration: Flow-through Fish Test” (OECD TG 305)
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): T-7701 (Pentafluoropropionic Acid), CAS# 422-64-0
- Substance type: Purchased from ABCR
- Physical state: Liquid
- Analytical purity: 97%
- Stability under test conditions: The test substance was demonstrated to be stable under storage and
test conditions by the analysis of samples collected before, during, and after the definitive test.
- Storage condition of test material: Cold and dark storage place.
Radiolabelling:
no

Sampling and analysis

Details on sampling:
- Details on sampling and analysis of test organisms and test media samples:

Analysis of the test substance in test water was conducted once before the first analysis of test fish and at the same time as that for test fish thereafter.

Analysis of test fish was performed 5 times at each exposure concentration during the exposure period. Four fish were removed at each sample period. Each analytical sample was made by combining tissues from two fish. This was necessary due to the small size of the fish

Lipid content was measured using three sets of two each control fish taken prior to the testing and at the termination of the testing period.

Sample frequency:
Water: after 1, 6, 11, 14, 21, and 28 days
Fish: after 6, 11, 14, 21, and 28 days

Test solutions

Vehicle:
no
Details on preparation of test solutions, spiked fish food or sediment:
Standards:

Test water: A standard curve was generated using test material as supplied by Sponsor. A stock solution of 1000 mg/L in ion-exchanged water was prepared. From the stock solution, standard solutions of 0.0500, 0.100, and 0.200 mg/L were prepared in deionized water (city water treated by Ultra pure water system) and analyzed by LC/MS. Peak areas under the mass fragmentograms (m/z 162.88) were linear with the corresponding mass of test substance in solution. The lowest detectable concentration of test substance was determined to be 0.0043 mg/L by the methods used.

Test fish: A standard curve was generated using test material as supplied by Sponsor. A stock solution of 1000 mg/L in ion-exchanged water was prepared. From the stock solution, standard solutions of 0.0500, 0.100, and 0.200 mg/L were prepared in acetonitrile / deionized water (city water treated by Ultra pure water system) (1/19 V/V) and analyzed by LC/MS. Peak areas under the mass fragmentograms (m/z 162.88) were linear with the corresponding mass of test substance in solution. The lowest detectable concentration of test substance was determined to be 0.0044 mg/L by the methods used.

Recovery and Test Blank:
All Recovery and Test Blank analyses were performed in duplicate.

Test blank: Dilution water revealed no detectable level of test substance (< 0.0043 mg/L).

Recovery from test water: Recovery of the test substance in water was taken to be 100% because the only pretreatment was dilution.

Recovery from fish: Portions of test substance (100 µg) were added to duplicate 10 g samples of chopped fish tissue, and subsequently subjected to preparation and analysis by LC/MS. Average recovery of test substance from samples containing fish tissue was 94.2%. It was assumed that when fish tissue was analyzed, recovery of the test substance accumulated in the experimental fish was similar to this value. Therefore, this fraction was applied to the quantitative analysis of test substance recovered when calculating bioconcentration factors.

PREPARATION AND APPLICATION OF TEST SOLUTION:

Stock solutions: Stock solutions of PFPA (T-7701) were prepared in deionized water (nominal concentrations; not adjusted for purity).
Level 1: 400 mg/L
Level 2: 40 mg/L

Test organisms

Test organisms (species):
Cyprinus carpio
Details on test organisms:
TEST ORGANISM
Test species: Cyprinus carpio (Carp)
Test fish age: Yearling
Length and weight: 7.7-9.6 cm; estimated to be ≈10 g/fish
Lot No. TFC-020221-II
Supplier: Fukuoka Fisheries & Marine Technology Research Center
Freshwater Laboratory
2449 Yamada, Asakura-machi, Asakura-gun, Fukuoka 838-1306, Japan

Pretreatment:

(1) External disinfection in flow-through tanks using 20 mg/L ELBAZIU and 7 g/L NaCl for 24 h. Fish were subsequently held 81 days in a flow-through system at 25 + 2 ºC.

(2) Fish were transferred to the acclimatizing aquarium, where they underwent a 24-h external disinfection with 50 mg/L oxytetracycline hydrochloride (OTC for fisheries) and 7 g/L sodium chloride. Fish were held in the acclimatizing aquarium under flow-through conditions for 53 days.

(3) Fish were transferred to test tanks where they underwent a final 24-h external disinfection as described in Pretreatment (1). The fish were held in the test tanks (flow-through systems) for 15 days before beginning exposure to the test substance.

Feeding: Feed for fry of carp, Nippon Formula Feed Mfg. Co., Ltd.
Amount equivalent to approximately 2% of total body weight was divided in half and fed to the fish in two portions each day. Prior to each sampling, the fish were starved for 24 h.

Study design

Route of exposure:
aqueous
Test type:
flow-through
Water / sediment media type:
natural water: freshwater
Total exposure / uptake duration:
28 d

Test conditions

Hardness:
83.5 mg/L (Ca, Mg)
Test temperature:
25.0 – 25.4 ºC (recorded once per day).
pH:
8.1 ± 0.1 (recorded once per week or more).
Dissolved oxygen:
7.9 – 8.1 mg/L (recorded twice a week).
TOC:
0.3 mg/L (based on dilution water quality testing)
Salinity:
NA
Details on test conditions:
Test water: Groundwater from premises of Kurume Laboratory

Water chemistry: Total analysis performed twice yearly; dilution water sampled on 05/09/2002 for total analysis. See attachment titled "Water Quality Analysis" under heading "Any other information on materials and methods incl. tables".

Exposure vessels: 100-L tank, flow through

Flow rate of test water: 2 mL/min for stock solution
800 mL/min for dilution water
1,155 L/day of test water supplied
Flow rate tested once per day utilizing a graduated cylinder

Number of fish / vessel: 29 (Level 1 and Level 2)
20 (Control)

Cleaning of test tank: Approximately once per day.

Light cycle: 14 h light/day; white fluorescent lamp

RANGE-FINDING / PRELIMINARY STUDY

Method/Guideline followed: JIS K 0102-1998-71 ”Testing methods for industrial waste water, Acute toxicity test with fish.”

GLP (Y/N): Yes

Year: 2002

Test species: Oryzias latipes (Orange-red killifish; Japanese medaka)
Lot TFO-020426

Test fish age: Not reported

Length and weight: 3.3 cm; 0.31 g

Supplier: Ogawa shoten
181 Ooishi-machi Kurume-shi, Fukuoa 830-0049, Japan

Analytical monitoring: None noted.

Exposure period: 96 h

Exposure concentrations: One Negative Control, Test sample concentrations of 98.8, 148, 222, 333, and 500 mg/L (as 100% test sample. Does not adjust for purity of target substance.).
NOTE: Confirmation of expected sensitivity of the test organism was noted by exposing the same rearing (Lot TFO-020426) to a reference test substance (pentachlorophenol sodium salt) which yielded a 48-hour LC50 of 0.642 mg/L.

Exposure vessels: Round glass vessel each containing 4 L of test solution

Loading: 0.78 g fish / L

Number of fish / vessel: 10

Pretreatment: External disinfection in flow-through tanks using 20 mg/L ELBAZIU and 7 g/L NaCl for 24 h; disinfection repeated after removal of abnormal fish. Acclimatization was performed in a flow-through system for 67 days at 25 ± 2 ºC.

Test Conditions:

Test water: Groundwater from premises of Kurume Laboratory

Temperature: 25.0 to 25.3 ºC

Water chemistry: Total analysis performed twice yearly; dilution water removed 05/09/2002.

Total hardness: 83.5 mg/L (Ca, Mg)
pH: 7.9
Conductivity: 459 µS/cm (µmho/cm)

Stock and test solution preparation: Stock solutions of test substance were prepared in deionized water at a nominal concentration of 1000 mg/L. This was diluted as appropriate into the test water.

Test type: Semi-static, with complete test water exchange every 8 – 16 h.

Water chemistry during the study:
Dissolved oxygen: 8.1 mg/L at initial exposure
6.1 - 8.0 before renewal of test water
pH: 2.9 – 7.2 at initial exposure
2.9 – 7.5 before renewal of test water

RESULTS

Concentration Cumulative Mortality (%)
(mg/L) 24 h 48 h 72h 96 h
Control (0) 0 0 0 0
98.8 0 0 0 0
148 0 0 0 0
222 0 0 0 0
333 0 0 0 0
500 100 100 100 100

The value of the LC50 was estimated using a log-linear plot of the data
LC50 = 408 mg/L

CONCLUSIONS

The 96-hour LC50 for perfluoropropionic acid (T-7701) in Oryzias latipes is estimated to be 408 mg/L.

Submitter: 3M Company, Environmental Laboratory, 3M Center, Environmental, Health and Safety Operations, Building 0260-05-N-17, Maplewood, MN 55144-1000

DATA QUALITY

Reliability: Klimisch ranking 2. This was a generally well conducted study. No report of the number of replicate tests was made, so it must be assumed that only one replicate test at each concentration of the test substance was performed. Mortality at the highest concentration may be a direct result of the pH being low (pH = 2.9). Had pH been buffered, mortality may not have occurred.
Nominal and measured concentrations:
Exposure concentrations: 1.00 and 0.100 mg/L (nominal)
Measured: Level 1 = 0.918 ± 0.0315 mg/L (average ± standard deviation)
Level 2 = 0.0923 ± 0.00907 mg/L (average ± standard deviation)
Reference substance (positive control):
no

Results and discussion

Lipid content
Lipid content:
3.97 - 4.31 %
Time point:
other:
Bioaccumulation factor
Type:
BCF
Value:
>= 1.2 - <= 4.8
Basis:
whole body w.w.
Remarks:
BCF 1.2, 1.00 mg/L exposure. BCF <=4.8, 0.100 mg/L exposure
Time of plateau:
11 d
Calculation basis:
steady state
Remarks on result:
other: Conc.in environment / dose:Nominal:    1.00 and 0.100 mg/L. Details on conc. in environment / dose --> see details on results
Depuration
Elimination:
not specified
Details on kinetic parameters:
None given.
Metabolites:
None noted.
Results with reference substance (positive control):
None.
Details on results:
Details on conc. in environment dose level :
Measured: Level 1 = 0.918 ± 0.0315 mg/L (average ± standard deviation)
Level 2 = 0.0923 ± 0.00907 mg/L (average ± standard deviation)

Loading: ~2.9 g fish / L for Test vessels (at beginning of study)
~2.0 g fish / L for Control vessel (at beginning of study)

Neither mortality nor abnormalities in behavior were observed in treated and control fish during the 28 day test period.

The report indicates that the concentration of PFPA in fish exposed to Level 2 (0.100 mg/L nominal), could not be quantified at the Limit of Quantitation (LOQ). Thus, all results for BCF at level 2 exposure are based on the result being less than the quantifiable level.
Reported statistics:
Not given.

Any other information on results incl. tables

Remark lipid content:

Lipid content (LO) represents the average lipid content at termination of testing. Lipid content (HI) represents the average lipid content prior to testing. Each value represents an average of three determinations, each using combined tissue from two fish.

Lipid content was measured using three sets of two each control fish taken prior to the testing and at the termination of the testing period. The fish were measured and weighed, minced, and homogenized as is. A weighed portion of the homogenate was extracted with chloroform – methanol (ratio not specified), and the residue after solvent removal measured gravimetrically. Results represent the average of three determinations at each of the two time points.

Concentrations of test item in test water
(Unit: mg/L)
Level After 1 day After 6 days After 11 days After 14 days After 21 days After 28 days Average (Stndard deviation)
1 0.913 0.970 0.911 0.930 0.873 0.911 0.918 (0.0315)
2 0.0763 0.102 0.0928 0.0982 0.0882 0.0963 0.0923 (0.00907)

Concentration of test substance in fish
(1 mg/L nominal exposure)
Exposure
(days)
Replicate Concentration in fish*
(ug/g)
6 a 0.877
b <LOQ
11 a <LOQ
b 1.08
14 a 1.72
b <LOQ
21 a 1.34
b 0.916
28 a 1.13
b 1.05
Concentration of test substance in fish
(0.1 mg/L nominal exposure)
Exposure
(days)
Replicate Concentration in fish*
(ug/g)
6 a <LOQ
b <LOQ
11 a <LOQ
b <LOQ
14 a <LOQ
b <LOQ
21 a <LOQ
b <LOQ
28 a <LOQ
b <LOQ
* Limit of quantitation in fish = 0.47 ug/g

Bioconcentration Factors
Level After 6 days After 11 days After 14 days After 21 days After 28 days
1 0.93
≤0.51*
≤0.51*
1.2
1.8
≤0.51*
1.5
1.0
1.3
1.2
2 ≤5.0*
≤5.0*
≤5.0*
≤5.0*
≤5.0*
≤5.0*
≤5.0*
≤5.0*
≤5.0*
≤5.0*
* Results based on concentration in fish below limit of quantitation.

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
PFPA does not bioconcentrate in carp under the described test conditions.
Executive summary:

In a 28-day bioconcentration study, carp (Cyprinus carpio) were exposed to PFPA at nominal and measured concentrations of 1 and 0.1 mg/L, 0.918 (+0.0315) and 0.0923 (+0.00907) mg/L respectively under continuous flow through conditions.  The 28 day BCF was 1.2 and<4.8 for the 1 and 0.1 mg/L exposures respectively. No sublethal effects were observed in the test groups.  Based on the results of this study, PFPA would not be classified as bioaccumulative in Carp.

 

While a few data points are missing (e.g. report of actual fish weight, descr. of behavioral patterns observed for determination of “abnormalitites in behavior”) the quality of the data reported and conclusions are not diminished. This bioconcentration study is classified as acceptable and satisfies the guideline requirement for Bioconcentration study in Carp.

 

Results Synopsis

 

Test organism size/age: Yearling carp, 7.7 - 9.6 cm; estimated to be ~10 g/fish

Test Type: Flowthrough

 

BCF Level 1 (1 mg/L):  1.2

BCF Level 2 (0.1 mg/L): <4.8