Document 4vOarxJRvQXJQ99pRmDQqzrBp
RECEIVED OPPTNCiC
2003 MAR 10 AH II** 5 I
Telomer Research Program
1200 South Hayes Street Arlington, Virginia 22202-5050 Tet (703)413-1100 ad.5264 Fax: (703)4136111
M &Q& - I AW
Clariant
February 14, 2003
Mary F. Dominiak U.S. Environmental Protection Agency EPA East (Mail Code 7405M) 1200 Pennsylvania Avenue NW Washington, DC 20460 E-mail: dominiak.mary@epa.gov
Dear Mary:
The Telomer Research Program (TRP) and its member companies, Asahi Glass, Clariant, Daikin, and DuPont, are providing to U.S. EPA Docket# AR226 final reports on the acute hazard of 8-2 Telomer B Alcohol (3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecaf!uorodecan-1-ol; CAS No. 678-39-7) when tested with standard freshwater aquatic test species. These reports present the results of the acute aquatic toxicity testing conducted by an independent laboratory at the request of the TRP. These studies were conducted according to existing OECD test guidelines and in accordance with the provisions of the appropriate Good Laboratory Practices (GLP) regulations.
The physical-chemical characteristics of the test substance indicate that the test substance is a "difficult" substance, as indicated in OECD Monograph 23 (Guidance Document on Aquatic Toxicity Testing of Difficult Substances and Mixtures, OECD 2000), and the experimental designs selected for the acute aquatic toxicity testing were reflective of this observation. Common practices for testing these types of materials, and for evaluating the results from tests of these types of materials, were used during the conduct of the studies and the preparation of the study reports.
The TRP and Its member companies believe that the data gathered from these studies are consistent with the hypothesis that the 8-2 Telomer B Alcohol is of low to medium concern in terms of its potential for causing acute effects on aquatic species. If there are questions concerning the technical aspects of the studies submitted, please contact me so that I may direct your questions to the appropriate TRP member company personnel.
Sincerely,
Katie u. smythe TRP Administrator
This document does not contain TSCA CBI.
000043
Dr.U.Noack-Laboratorium fr Angewandte Biologie
1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro-
Alga, Growth Inhibition Test with Scenedesmus subspicatus, 72 h, Limit-Test with the Water Accommodated Fraction (WAF)
acc. to OECD-Guideline No. 201 for Testing of Chemicals (June 1984)
Sponsor Telomer Research Project
Asahi G lass At o f in a C la r ia n t
D a ik in In d u s t r ie s Ltd Dupont
Author Dirk Scheerbaum
Test Facility
D r .u .N OACK- La b o r a t o r iu m FR ANGEWANDTE BIOLOGIE
Kthe-Paulus-Str. 1 D-31157 Sarstedt
Laboratory Project ID
Project-No.
020109CK
Study-No.
S S 084112
Study Completed On August 07, 2002
Page 1 of 39
Original 7 of 7
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Page 2 of 39
1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro-
Alga, Growth Inhibition Test with Scenedesmus subspicatus, 72 h, Limit Test Project-No.020109CK
with the Water Accommodated Fraction (WAF), acc. OECD 201_____________ Study-No. SSQ84112
Statement of GLP Compliance
T itle
Guideline Test Item Testing Facility
Deviations from GLP Principles
1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoroAlga, Growth Inhibition Test with Scenedesmus subspicatus, 72 h, Limit-Test with the Water Accomodated Fraction (WAF)
OECD-Guideline No. 201 for Testing of Chemicals (1984)
1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro(Batch number: P00/001)
D r .U .N o a c k -L a b o r a t o r iu m f r a n g e w a n d t e B io l o g ie
Kthe-Paulus-Str.1, D-31157 Sarstedt
Phone: (+49) 050 66/706 70, Fax: (+49) 05066/706 789, E-mail: info@noacK-lab.de
None
We declare that this study was conducted and reported in compliance with the actual OECD principles of Good Laboratory Practice and the national GLP regulations as specified in the law in force, except deviations mentioned above.
August 07, 2002 August 07, 2002 August 07, 2002
(Dirk Scheerbaum, Study Director) (Thomas Geffke, Scientist) (Dr. Udo Noack, Head of Testing Facility)
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1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro-
Alga, Growth Inhibition Test with Scenedesmus subspicatus, 72 h, Limit Test Project-No.020109CK
with the Water Accommodated Fraction (WAF), ace. OECD 201_____________ Study-No. SSQ84112
Statement of the Quality Assurance Unit
Title
Guideline Test Item Study Director
1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoroAlga, Growth Inhibition Test with Scenedesmus subspicatus, 72 h, Limit-Test with the Water Accomodated Fraction (WAF)
OECD-Guideline No. 201 for Testing of Chemicals (1984)
1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro(Batch number: P00/001)
Dirk Scheerbaum
The study was verified as follows:
inspection study plan process based
report
dates
2002-03-12
2002-04-02
2002-04-07 2002-04-11 2002-04-12 2002-07-15
date of report
2002-03-12
2002-04-02
2002-04-08 2002-04-11 2002-04-12 2002-07-15
The reported results accurately and completely reflect the raw data of the study. Also methods, procedures, and observations are accurately and completely described in the report.
The accordance of the study with its study plan and the principles of Good Laboratory Practice is guaranteed.
August 07, 2002
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1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro-
Alga, Growth Inhibition Test with Scenedesmus subspicatus, 72 h, Limit Test Project-No.020109CK
with the Water Accommodated Fraction (WAF), acc. OECD 201_____________ Study-No. SSQ84112
Personnel Involved
Study Director:
Dirk Scheerbaum (Biologist)
Scientist:
Thomas Geffke (Chemical Engineer) Responsible for concentration control analysis
Technical Staff:
Ute Kutzner
Quality Assurance Unit:
Gudrun Mhrmann-Kalabokidis (Biologist)
Deputy:
Susanne Becker (Biologist)
Head of Testing Facility:
Dr. Udo Noack (Biologist)
000047
Dr.U.Noack-Laboratorium fr Angewandte Biologie
Report 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoroAlga, Growth Inhibition Test with Scenedesmus subspicatus, 72 h, Limit Test with the Water Accommodated Fraction (WAF), acc. OECD 201
Page 5 of 39
Project-No.020109CK Study-No. SS084112
Table of Content
Title Page Statement of GLP Compliance Statement of Quality Assurance Unit Personnel Involved Table of Content List of Tables List of Figures
1 2 3 4 4 .1 4 .2 4 .3 5 6 7 8 9 10
10.1 1 0 .2 1 0 .2 .1 10 .2.2 10 .2.3 10 .2.4 10 .2.5 1 0 .3 1 0 .4 1 0 .5 11 12
13
Summary Characterisation Data of the Test Item Method Results Preliminary Test Definitive Test Test of the Reference Item Validity Criteria Conclusions Literature / References Graph Physicochemical Data G C -A n a ly s is of 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9, 10,10,10heptadecafluoroMethod Method Validation Linearity Quantification Limit and Detection Limit Precision (Repeatability) Peak Symmetry Accuracy Results Calibration Curves Chromatograms Raw Data Certificate of 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10heptadecafluoro- Analysis GLP-Certificate o f D r .U .N o a c k -L a b o r a t o r iu m
Page
1
2
3
4
5
6
6
7 9 10 17
17
17 20 20 21 21 22 23
24 24 26 26 26 27 27 28 29 30 33 36
38 39
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Report
1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoroAlga, Growth Inhibition Test with Scenedesmus subspicatus, 72 h, Limit Test with the Water Accommodated Fraction (WAF), acc. OECD 201
Page 6 of 39
Project-No.020109CK Study-No. SS084112
List of Tables
Tab. 1
Tab. 2: Tab. 3
Tab. 4 Tab. 5 Tab. 6
Tab. 7 Tab. 8 Tab. 9 Tab. 10.
NOEC/LOEC and ECS0-Values (0-72 h) of Biomass Inhibition and Rate-Related Inhibition Preliminary Test Results: Cell Density Values NOEC/LOEC and EC50-Values (0-72 h) of Biomass Inhibition and Rate-Related Inhibition Cell Densities Evaluation after 72 h ECso-Values of Biomass Inhibition and Rate-Related Inhibition of the Reference Item Water Parameters Repeatability of Injections of Test Item Mean Recovery Rate of Fortified Samples Concentrations of the Active Ingredient 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro-
Page
8 17
18 18 19
20 23 27 28
29
List of Figures
Fig. 1 Fig. 2 Fig. 3 Fig. 4 Fig. 5 Fig. 6 Fig. 7
Cell Densities for Control and Limit Concentration (0-72 h) Calibration Curve of Standard - concentration range 0.01 - 0.06 mg/L Calibration Curve of Standard - concentration range 0.1 - 10 mg/L Calibration Curve of Standard - concentration range 0.1 - 0.6 mg/L Chromatogram of Standard Chromatogram of Test Item Chromatogram of Control
22 30 31 31 33 34 35
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1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro-
Alga, Growth Inhibition Test with Scenedesmus subspicatus, 72 h, Limit Test Project-No.020109CK
with the Water Accommodated Fraction (WAF), acc. OECD 201_____________ Study-No. SSQ84112
1 Summary
The acute toxicity of 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro- (batch number P00/001) to the unicellular freshwater green alga Scenedesmus subspicatus was determined according to the principles of OECD-Guideline 201, at D r .U .N o a c k -L a b o r a t o r iu m F r A n g e w a n d t e B io l o g ie in 31157 Sarstedt, Germany from April 01 to 04, 2002.
The study was conducted under static conditions over a duration of 72 hours with an initial cell density of nominally 104 cells/mL and a water accommodated fraction as limit concentration, containing 0.47 mg/L. For the preparation of the water accommodated fraction (WAF) a suspension with a nominal loading of 100 mg/L was shaken for 96 h.
Test vessels were coated with the test item for 14 days before the test started. Three replicates were tested for the limit concentration and 6 replicates for control. Microscopic evaluation of the cells at the start and end of the incubation period revealed no morphological abnormalities. W ater quality parameters of pH-value, measured at 0 and 72 h, and room temperature, measured continuously, were determined to be within the acceptable limits.
The concentration of the active ingredient 1-Decanol, 3,3,4,4,5,5,6,6, 7,7,8,8,9,9,10,10,10heptadecafluoro- was analysed at the limit concentration level at test start and test end. Details of the analytical results are given in Part 10. The initially measured concentration of 1Decanol, 3,3,4,4,5,5,6,6, 7,7,8,8,9,9,10,10,10-heptadecafluoro- was 0.47 mg/L. At the end of the test the concentration of the test item was below the detection limit of 0.009 mg/L. Sorption to the glass walls of the test vessels is unlikely as the walls were already pre-coated for 14 days before the test has started.
The effects of 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro- based on initially measured a.i. concentration are summarized in Table 1.
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1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,1O-heptadecafluoro-
Alga, Growth Inhibition Test with Scenedesmus subspicatus, 72 h, Limit Test Project-No.020109CK
with the Water Accommodated Fraction (WAF), acc. OECD 201_____________ Study-No. SSQ84112
Tab. 1: NOEC/LOEC and ECgo-Values (0-72 h) of Biomass Inhibition and Rate-Related Inhibition N O E C /L O E C calculated using A N O V A (O n e W a y A nalysis of V arian c es) and Dunnett's
test (oc=0.05), b a s e d on in itially m easu re d co n ce n tratio n s [m g/L ] .
EbCso 95 % confidence interval LOEC NOEC
ErCso 95 % confidence interval LOEC NOEC
Inhibition of biomass growth [mg/L] > 0.47
Not applicable > 0.47 0.47
Rate-related inhibition [mg/L] > 0.47
Not applicable > 0.47 0.47
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Report 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro-
Page 9 of 39
Alga, Growth Inhibition Test with Scenedesmus subspicatus, 72 h, Limit Test Project-No.020109CK
with the Water Accommodated Fraction (WAF), acc. OECD 201_____________ Study-No. SSQ84112
2 Characterisation Data of the Test Item
TEST ITEM
1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro-
Batch number
P0/001
CAS RN
678-39-7
Chemical characterization / 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluorodecane-l-ol
Active ingredient
(EINECS)
Purity
C8-perfluoroalkylethanol 99.3 % (ala), sum perfluoroalkylethanol 99.9 % (a/a)
Appearance
Yellowish solid wax
Water solubility
134 pg/L at 12 degree C, Std 29 pg/L, n=8 (DuPont, 2002) 137 pg/L at 25 degree C, Std 53 pg/L, n=17 (DuPont, 2002)
Stability in water Density
Stable at test conditions Ca. 1.7 g/cm3
Expiry date Recommended storage Storage at testing facility Retention Identification parameter at testing facility
Unlimited if stored under the recommended conditions. Room temperature (20 C), protected from moisture. Room temperature, protected from moisture and light. At least 1 g has been retained.
Name, batch number and colour.
The test item and the information concerning the test item were provided by the sponsor.
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Page 10 of 39
1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro-
Alga, Growth Inhibition Test with Scenedesmus subspicatus, 72 h, Limit Test Project-No.020109CK
with the Water Accommodated Fraction (WAF), acc. OECD 201_____________ Study-No. SSQ84112
3 Method
TEST GUIDELINE
OECD-Guideline No. 201 for Testing of Chemicals (adopted 7June, 1984)
TYPE AND PURPOSE OF THE STUDY
Alga, growth inhibition test for determination of the effects on growth rate and biomass production with a water accomodated fraction (WAF) of the test item over a period of 72 h (Limit Test).
TEST SYSTEM Test organism Reason for the selection of the test system Origin
Date of receipt Method of cultivation
Culture medium
Scenedesmus subspicatus CHODAT SAG 86.81
According to the guideline Scenedesmus subspicatus CHODAT SAG 86.81 is suitable for this kind of study.
Sammlung von Algenkulturen (SAG), Pflanzenphysiologisches Institut der Universitt Gttingen, Nikolausberger Weg 18, D-37073 Gttingen
1997-04-07
Fresh stocks were prepared every month on Z-Agar. Light intensity amounted 35-70 pE/m2 s for 24 h per day.
Nutrient medium Z according to L t t g e et al. (1994).
TEST ITEM
Pretreatment of the test item
1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10heptadecafluoro-
The test item was heated to 50 "C before weighing for homogenisation.
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1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro-
Alga, Growth Inhibition Test with Scenedesmus subspicatus, 72 h, Limit Test Project-No.020109CK
with the Water Accommodated Fraction (WAF), acc. OECD 201_____________ Study-No. SSQ84112
Coating of the test vessels
A coating phase of 14 days was carried out.
Preparation
200 mg 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10heptadecafluoro-/L dilution water (total volume 2 L) were shaken in a rotating shaker with 20 rpm for 96 h at 20 C. 2 L -brow n glass flasks were used. After phase separation 100 mL per test replicate was transferred from the clear liquid phase to 250 mL Erlenmeyer flasks and closed with plugs. Dilution water was used for the preparation of the coating solution.
Coating period
The readily filled and closed Erlenmeyer flasks were stored at nominally 23 2 C for 14 days. At the end of the coating period the flasks were emptied and used for the test.
Preparation of the test
medium (water accomodated
fraction, WAF)
200 mg 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-
heptadecafluoro- /L dilution water (total volume 2 L) were shaken
in a rotating shaker with 20 rpm for 96 h at 20 C.
2 L -brow n glass flasks were used. After phase separation 100
mL per test replicate were transferred from the clear liquid phase
(representing the WAF) to the coated Erlenmeyer flasks.
CONTROL
Six replicates (without test item) were tested under the same test conditions as the test replicates.
SELF FLUORESCENCE
No self fluorescence was found at the water accomodated fraction.
REFERENCE ITEM
Potassium dichromate p.a. (Merck)
CAS No.
7778 - 50 - 9
Purity
100.2 %
Batch No.
K 233 78864646
Test concentrations (nom.) 0.1 - 0.2 - 0.4 - 0.8 - 1.6 - 3.2 mg/L
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1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro-
Alga, Growth Inhibition Test with Scenedesmus subspicatus, 72 h, Limit Test Project-No.020109CK
with the Water Accommodated Fraction (WAF), acc. OECD 201_____________ Study-No. SSQ84112
Range of validity Experimental date
0.20 mg/L < EbC60 (72 h) < 0.75 mg/L 0.60 mg/L < ErC60 (72 h) < 1.03 mg/L
November 27 to 30, 2001
TEST CONDITIONS Performance of the test Exposure duration Replicates Test container Test volume Dilution water
Preculture
Static procedure Experimental start date: April 01, 2002 Experimental completion date: April 04, 2002 Three replicates for limit concentration level, 6 per control Erlenmeyerflasks, 250 mL, with cotton wool plugs 100 mL According to the guideline:
Component NHCI
MgCI2 6 H20 CaCI2 2 H20 M gS O i 7 H20
KH2PO4 FeC h 6 H20 Na2EDTA 2 H20
H3 BO3 MnCI2 4 H20
ZnCI2 Na2MoO< 2 H20
CoCI2 6 H20 CuCI2 2 H20
NaHCOs pH
Concentration [mg/L]
15 12 18 15 1.6 0.08 0.1 0.185 0.415 3 X 10'3 7 x 10'3 1.5 x 10'3 1 X 10-s 50 8.2 0.2
A four day old preculture was used as inoculum. Incubation was performed in 500 mL Erlenmeyer flasks with dilution water. For the start of the test the preculture was diluted with dilution water to receive an initial cell concentration of approximately 1 x 10* cells/mL in the replicates. All algae were from the same source and have not been used in a previous study.
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1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro-
Alga, Growth Inhibition Test with Scenedesmus subspicatus, 72 h, Limit Test Project-No.020109CK
with the Water Accommodated Fraction (WAF), acc. OECD 201_____________ Study-No. SSQ84112
Application
Temperature Agitation
Light intensity Light regime
At the test start fluorescence was measured after application of the test item.
Nominally 23 2 C
Test containers were placed on a rotary shaker and oscillated at approximately 70 rpm.
Nominally 60 - 120 pE/m2 s
24 h/d light
KIND AND FREQUENCY OF MEASUREMENT
Equipment
Cell density was measured via Chlorophyll-a-fluorescence (excitation at 435 nm, emission at 685 nm). Each replicate was measured 6-fold. The cell density was measured at the beginning of the test and every 24 h. Filtrated dilution water was used as ground signal.
The pH-value at the beginning of the test was measured out of one additional replicate of the limit concentration and control. At the end it was measured from a pool of all replicates. The room temperature was measured continuously by a hygrothermograph. Light intensities were measured prior to test start.
Microscopic evaluation of the cells was determined at the start and end of the incubation. Alga cells were checked for any unusual cell shapes, color differences, differences in chloroplast morphology, flocculations, adherence of algae to test vessels or aggregation of alga cells.
pH-Meter, pH 191 (WTW)
H yg ro -th erm o g rap h e (K l im a t h e r m )
Chlorophyll-impulsfluorometer (K l e in f e l d ) Shaker, SM 25 A (BOhler) Centrifuge: Z K 364, Z ir b u s Inverse microscope ( Z e is s IM 2 5 ) Quantum Sensor ( S k y e In s t r u m e n t s Lt d ) Rotating shaker 3040, GFL Fluorescent tubes, O s r a m L 36 W / 11-860, daylight (test) Fluorescent tubes, Lampi 18 W, 60 cm, warmwhite (culture)
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1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro-
Alga, Growth Inhibition Test with Scenedesmus subspicatus, 72 h, Limit Test Project-No.020109CK
with the Water Accommodated Fraction (WAF), acc. OECD 201_____________ Study-No. SSQ84112
DETERMINATION OF THE TEST ITEM
The test concentration level and control was analytically verified by gas chromatography with mass spectrometric detection (m/z = 463 and 395) at test start and test end. At the end of the test replicates with alga were analysed. All details of the analytical method are given in part 10.
COURSE OF THE STUDY
Based on the results of a preliminary range finding test (NONGLP), the definitive test was performed with 1 limit concentration level with 3 replicates.
A control with dilution water (without test item, six replicates) was tested under the same conditions as the test group.
A four day-old preculture incubated at study conditions was used for the main study.
Chlorophyll-fluorescence was determined at the beginning and after 24, 48 and 72 h.
Other measurements were performed as specified above.
CALCULATIONS AND STATISTICS
Cell densities for each replicate were calculated from the fluorescence values 3-6 (see raw data). The fluorescence values were related to cell density values as follows:
Calculation of cell density out of fluorescence values
x = 63 Fq + 3625
x = cell density cells/mL
63 = factor cells/mL/ral.units/10 mL Fq = mean fluorescence value of all replicates per concentration
rel.units/10 mL
3625 = cell density cells/mL
Correlation dated April 12, 2001, r2 = 0.9963, n=8
The growth rate, the inhibition of biomass and the rate related inhibition after 72 h were calculated from the cell density values according to the formulas listed below.
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1-Oecanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro-
Alga, Growth Inhibition Test with Scenedesmus subspicatus, 72 h, Limit Test Project-No.020109CK
with the Water Accommodated Fraction (WAF), acc. OECD 201_____________ Study-No. SSQ84112
Calculation of biomass integral
N, - N0
N, + N2 - 2 N,,
N,,., * N , - 2 N ,
A " ------------ t , + ------------------------ (t2 - ti) * ... + ------------------------ (ln-U.1)
22
2
A = Area
No = nominal number of cells/mL at time to
Ni = measured number of cells/mL at time ti
Nn = measured number of cells/mL at time t,,
ti = time of first measurement after beginning of test
tn = time of nth measurement after beginning of test
(1)
Calculation of biomass inhibition
lA = ((Ac - AT) / Ac) 100 %
(2)
I* = biomass-inhibition %
Ac = biomass-integral in the control after n d in cells/mL
At = biomass-integral in the test concentration after n d in cells/mL
Calculation of growth rate
m = (In (N,,) - In (N0)) / (tn - 10)
U = growth rate cell density/d Nn = biomass after tn d in cells/mL No = biomass at to in cells/mL to = time of beginning of test t,, = time of n,h measurement after beginning of test
(3)
Calculation of rate related inhibition
lMt = ((Me - Mt) / Me) 100%
Iui = rate related Inhibition % pc = growth rate of the control after n d Ui = growth rate of the test concentrations after n d
(4)
NOEC, LOEC and statistical analysis
One way analysis of variance (ANOVA) and D U N N ETT's test of growth rate and biomass integrals, respectively were carried out for the determination of statistically significant differences compared to control replicates. When running a one way analysis of variance a normality test and an equal variance test was done first. P-values for both normality and equal variance tests were 0.05. The a-value for ANOVA and D u n n e t t 's test (acceptable probability of incorrectly concluding that there is a difference) was a=0.05.
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1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro-
Alga, Growth Inhibition Test with Scenedesmus subspicatus, 72 h, Limit Test Project-No.020109CK
with the Water Accommodated Fraction (WAF), acc. OECD 201_____________ Study-No. SSQ84112
Software
VALIDITY CRITERIA
DEVIATIONS FROM THE GUIDELINE DEVIATIONS FROM THE STUDY PLAN REPORTING ARCHIVING
The data for the tables in this report were computer generated and have been rounded for presentation from the full derived data. Consequently if calculated manually based on the given data minor variations may occur from these figures.
Calculations were carried out using software - SigmaPlot rel. 2000 (2000), Spss Inc. - SigmaStat rel. 2.03 (1992-1997), Spss Inc. - Excel 2000 (2000), Microsoft Corporation - TOXSTAT rel. 3.4 (1994), Western Ecosystems Technology,
Inc., Cheyenne, WY 82001 & David D. Gulley, University of Wyoming, WY, USA.
The cell density had to increase at minimum 16-fold in the control replicates within 72 h. The temperature during the test had to be in the range of 21-25 C controlled at 2 C. The pH-value of the control replicates should not normally deviate by more than 1.0 units during the test.
None
None
The final report is issued as paper copy (6 originals) and an electronic record (1 original).
The following was retained in the archive of the test facility for the period as specified in the operative national GLP regulations: all raw data s tu d y p la n (Original 1 of 2) final report paper copy (Original 1 of 6) all records performed by the quality assurance programme
including master schedules sample of test item Microfilms will be retained in a safe-deposit by Volksbank Sarstedt, D-31157 Sarstedt.
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Page 17 of 39
1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro-
Alga, Growth Inhibition Test with Scenedesmus subspicatus, 72 h, Limit Test Project-No.020109CK
with the Water Accommodated Fraction (WAF), acc. OECD 201_____________ Study-No. SSQ84112
4 Results
4.1 Preliminary Test
The results of the preliminary test conducted February 25 to 28, 2002 are presented below:
Tab. 2:
Preliminary Test Results: Cell Density Values (2 replicates each, control: 4 replicates)
Dilution level
1:1 o f th e W a te r Accom m odated Fraction
1:10 of the W ater Accom m odated Fraction
Control
R e p lic a te No.
1 2 1 2
1
2 3 4
Cell density [cells/mL] 0 h 72 h
11169 10555 11847 11138 11626 11390 10587 11547
536133 606378 551883 585903 586848 598818 626065 593463
No self fluorescence was found at the water accomodated fraction
4.2 Definitive Test
Cell densities are listed in Table 4. Evaluations of biomass inhibition and rate-related inhibition are given in Table 5 (calculated out of cell density values). The growth inhibition effects of the test item are summarized in Table 3.
Microscopic evaluation of the cells at the start and end of the incubation period revealed no morphological abnormalities.
Water quality parameters of pH-value, measured at 0 and 72 h, and room temperature, measured continuously, met the guideline requirements.
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1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro-
Alga, Growth Inhibition Test with Scenedesmus subspicatus, 72 h, Limit Test Project-No.020109CK
with the Water Accommodated Fraction (WAF), acc. OECD 201_____________ Study-No. SSQ84112
Tab. 3:
NOEC/LOEC and ECso-Values (0-72 h) of Biomass Inhibition and Rate-Related Inhibition N O E C /L O E C ca lculated using A N O V A (O n e W a y A nalysis o f V a ria n c e s ) and Dunnett's
E b C S0 95 %
confidence interval
LOEC NOEC
ErCso 95 %
confidence interval
LOEC NOEC
Inhibition of biomass growth [mg/L] > 0.47
Not applicable > 0.47 0.47
Rate-related inhibition [mg/L] > 0.47
Not applicable > 0.47 0.47
Tab. 4: Cell Densities ___________B ase d on In itially M e a s u re d C o n cen tratio ns
Test item
R e p lic a te No.
Oh
Cell D ensity [cells/m L]
24 h
48 h
1
9642
39378
178797
0.47 mg/L
2
9248
32668
207241
3
9264
30715
185758
mv
9385
34254
190599
1
9468
28242
165535
2
9342
31770
180624
3
9390
33219
177253
Control
4
8602
32495
173190
5
9169
32904
189664
6
8854
32873
182010
mv
9138
31917
178046
mv = mean value
72 h 592203 625120 618505 611943 555505 598188 595038 631263 577398 637405 599133
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Page 19 of 39
1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro-
Alga, Growth Inhibition Test with Scenedesmus subspicatus, 72 h, Limit Test Project-No.020109CK
with the Water Accommodated Fraction (WAF), acc. OECD 201_____________ Study-No. SSQ84112
Tab. 5:
E valuation after 72 h
Statistically significant differences of biom ass integrals and grow th rates com pared to control values are m arked (+), not significant differences are m arked (-). N O E C was c a lc u la te d using A N O V A (O n e W a y A nalysis o f V a ria n c e s ), Dunnett's an d W illiam's test (a = 0 .0 5 ), based on initially m easured concentrations.
Test item
R e p lic a te
No.
0 .4 7 m g/L
1 2 3 mv
1 2
Control
3 4 5
6 mv
mv = mean value
Biom ass Integral
Inhibition of Biom ass
Growth R a te
490172 529349 502566 507362
[%] -0 .7 2 - 8.77 - 3.26 (-) -4 .2 5
1.37 1.40 1.40 1.39
447860 488133 484516 499812 488345 511451 486685
1.36 1.39 1.38 1.43 1.38 1.43 1 .3 9
negative inhibitions = Increase of growth
R ate-R elated Inhibition
[%] 1.56
-0 .7 3 -0 .4 3 (-) 0.13
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1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro-
Alga, Growth Inhibition Test with Scenedesmus subspicatus, 72 h, Limit Test Project-No.020109CK
with the Water Accommodated Fraction (WAF), acc. OECD 201_____________ Study-No. SSQ84112
4.3 Test of the Reference Item
The acute toxicity of potassium dichromate to the unicellular freshwater green alga Scenedesmus subspicatus was determined over a period of 72 h from November 27 to 30, 2001.
Tab. 6:
ECso-Values of Biomass Inhibition and Rate-Related Inhibition of the Reference Item based on nominal concentrations [m g/L], (0-72 h)
EbCso 95 % confidence interval
ErC5o 95 % confidence interval
Inhibition of biomass growth 0.49
0.45-0.54 Rate-related inhibition
0.94
0 .8 2 - 1.07
The EC50 - values of the reference item potassium dichromate after 72 h met the validity criteria according to EEC Directive 92/69/EEC Method C.3 Annex 2 (valid ranges are 0.20 0.75 mg/L and 0.60 - 1.03 mg/L potassium dichromate for inhibition of biomass growth and rate-related inhibition, respectively).
5 Validity Criteria
The study met the validity criteria of the guideline:
The cell growth increased 66-fold after 72 h in the control (required: 16-fold). The temperature during the test was in the range of 21-25 C. The difference from the mean was within 2 C: min.: 22.0 C, max.: 24.0 C mean value: 23.0 C. The pH-value in the control replicates increased not higher than 1.0 units: 8.13 to 8.54.
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1-Decanol, 3,3,4,4,5,5,6,6,7,7,818,9,9,1 0,10,10-heptadecafluoro-
Alga, Growth Inhibition Test with Scenadesmus subspicatus, 72 h, Limit Test Project-No.020109CK
with the Water Accommodated Fraction (WAF), acc. OECD 201_____________ Study-No. SSQ84112
6 Conclusions
In this study 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro- was not found to inhibit the growth of the freshwater green alga Scenadesmus subspicatus tested with the water accommodated fraction (WAF) containing 0.47 mg/L (initially measured concentration).
7 Literature / References
(1) DuPont, Determination of physico-chemical parameters of Perfluorooctylethanol, CAS No. 67839-7, Unpublished results, Update Feb. 15, 2002
(2) EEC Directive 92/69/EEC Method C.3, EEC L 383A/179 (December, 1992)
(3) LOt t g e , U, S c h n e p f , E., LAu c h l i, A., N a g a t a , T. (editors, 1994): Botanica Acta, Journal o f the German Botanical Society, N o . 3 Volume 1 0 7 p a g e 1 1 1 - 1 8 6 (June 1994), T h ie m e -
V erlag.
(4) OECD - guideline no. 201 for testing of chemicals: Alga, growth inhibition test. Adopted June 7, 1984
(5) OECD Series on Testing and Assessment, No 23 Guidance Document on Aquatic Toxicity Testing of Difficult Substances and Mixtures ENV/JM/Mono (2000)6.
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1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro-
Alga, Growth Inhibition Test with Scenedesmus subspicatus, 72 h, Limit Test Project-No.020109CK
with the Water Accommodated Fraction (WAF), acc. OECD 201_____________Study-No. SSQ84112
8 Graph
Fig. 1: Cell Densities for Control and Lim it Concentration (0-72 h)
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Page 23 of 39
1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro-
Alga, Growth Inhibition Test with Scenedesmus subspicatus, 72 h, Limit Test Project-No.020109CK
with the Water Accommodated Fraction (WAF), acc. OECD 201_____________ Study-No. SSQ84112
9 Physicochemical Data
Tab. 7: Water Parameters
Control Water Accommodated Fraction
pH-value
Start: 0 h
End; 72 h
8.13 8.07
8.54 8.08
Temperature: min.: 22.0 C max.: 24.0 C mean value: 23.0 C
The mean light intensity was 67.2 pE/m2 s (minimal 61.5, maximal 74.2 pE/m2 -s).
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Page 24 of 39
1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,1O-heptadecafluoro-
Alga, Growth Inhibition Test with Scenedesmus subspicatus, 72 h, Limit Test Project-No.020109CK
with the Water Accommodated Fraction (WAF), acc. OECD 201_____________ Study-No. SSQ84112
10 GC - Analysis of 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9, 10,10,10-heptadecafluoro-
10.1 Method METHOD OF DETERMINATION
Equipment
Analyses of various concentrations of 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluorowere carried out via gas chromatography on a Fused Silica CPSIL 8 capillary column using external standards. Detection was performed with a mass selective detector (MS).
Gas chromatograph Autosampler Detector Software
: CP-3800, V a r ia n : CTC Combi PAL, CTC A n a l y t ic s : MS, Saturn 2000, VARIAN : Saturn GC/MS Workstation 5 .5 1 , V a r ia n
Reagents
ferf.-butyl methyl ether 99.8 %, Batch 423370/1 31101, F l u k a
CONDITIONS OF ANALYSIS
Column
Injector Injector temperature Injection volume Temperature programme
Chrompack CP-SIL 8 (V a r ia n ), 30 m, i.d.: 0.25 mm, LOW BLEED / MS, Film thickness: 0.25 pm Splitless (30 psi pressure puls) for 1 min 250 C 1 pL
Heating rate [C/min]
--
10 30
Temperature [C] 40 90 220
Holding time [min]
0.0
1.0 5.0
Carrier gas Detector Ionisation mode
Run tim e R etention tim e
Helium 1 mL / min column flow MS Electron ionisation (El)
1 5 .3 3 m in a p p ro x. 4 .9 m in
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1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafiuoro-
Alga, Growth Inhibition Test with Scenadesmus subspicatus, 72 h, Limit Test Project-No.020109CK
with the Water Accommodated Fraction (WAF), acc. OECD 201_____________ Study-No. SSQ84112
STANDARD
PREPARATION OF STANDARDS
PREPARATION OF SAMPLES
Test item was used as standard.
Two stock solutions of the standard of 200 mg/L were prepared in ferf.-butyl methyl ether, diluted (9 concentrations each) and used for calibration. For calibration range see part 10.2.1.
40 mL of the sample were extracted with 4.0 mL ferf.-butyl methyl ether. After phase separation the extract was injected directly into the GC/MS.
METHOD VALIDATION Peak symmetry
Repeatability Accuracy
Quantification of the test item was calculated by peak area of the masses 69 + 395 + 463 (m/z) based on the external standards. The calibration procedure and the principle for the determination of detection and quantification limits were adapted from DIN 32645. The method was validated as described below:
Peak symmetries were calculated from chromatograms of analytical standards and test item according to the following equation:
T = Wo.os / (2-f)
T
Wo.05
f
= Peak symmetry
= Peak width at 5 % of peak height [mm]
= first sector of peak width ( divided by the perpendicular line from the peak m axim um ) [mm]
5 sub-samples of the highest and lowest concentration prepared from a single homogenous sample were analysed. Mean values, standard deviations and variations of coefficients were calculated.
5 samples with 40 mL water were spiked with an amount of 8 pg 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro(80 pL of 100 mg/L) to give a final concentration of 200 pg/L. Afterwards samples were extracted as described above.
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1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro-
Alga, Growth Inhibition Test with Scenedesmus subspicatus, 72 h, Limit Test Project-No.020109CK
with the Water Accommodated Fraction (WAF), acc. OECD 201_____________ Study-No. SSQ84112
10.2 M ethod V alidation
The method was validated and tested with satisfactory results with respect to linearity precision (repeatability) peak symmetry accuracy determination limit and detection limit.
10.2.1 Linearity
The analytical system gave linear response in the range of 0.1 - 10 mg/L of 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro-. The calibration curve and chromatograms obtained for standard and test item are given in figures 2 - 7 . For determination and detection limits see part 10.3.
10.2.2 Quantification Lim it and Detection Limit
The limit of quantification (LOQ) and limit of detection (LOD) were calculated using the blind value method described in DIN 32645 "Nachweis- und Bestimmungsgrenze" . The LOD was determined to be 0.009 mg/L and therefore the LOQ was 0.027 mg/L (threefold the LOD).
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1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro-
Alga, Growth Inhibition Test with Scenedesmus subspicatus, 72 h, Limit Test Project-No.020109CK
with the Water Accommodated Fraction (WAF), acc. OECD 201_____________ Study-No. SSQ84112
10.2.3 Precision (Repeatability) The results of the repeatability tests are provided in the table below.
Tab. 8:
R e p e a ta b ility o f In jectio n s o f Test Item Peak area [pV*s]
Serial No 1 2 3 4 5
Mean SD C V [%]
1.HorpnAl
3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10heptadecafluoro-
(m /z = 69+395+463)
0.1 mg/L
0.6 mg/L
2098 2098 1978 1958 2341 2094.6 1 5 2 .5
12141 13563 13291 13597 13500 13218.4 6 1 3 .9
7.3 4.6
10.2.4 Peak Symmetry
The peak symmetry of the test item 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10heptadecafluoro- was 1 .8.
000070
Dr.U.Noack-Laboratorium fr Angewandte Biologie
Report 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoroAlga, Growth Inhibition Test with Scenedesmus subspicatus, 72 h, Limit Test with the Water Accommodated Fraction (WAF), acc. OECD 201________
Page 28 of 39
Project-No.020109CK Study-No. SSQ84112
10.2.5 Accuracy
Tab. 9: Mean Recovery Rate of Fortified Samples
Fortified concentration
of test item
0 .1 9 8 6 [mg/L] Replicate 1
Replicate 2
Replicate 3
Replicate 4
Replicate 5
Mean SD (N-1) CV% LO Q [mg/L] LO D [mg/L] Linearity correlation coefficient ( r2 )
1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10
-heptadecafluoro-
calc. cone.
RR
[mg/L]
[%]
0.22
111
0.24 0.23
121 116
0.21 0.21
106 106
0.21 0.22
106 111
0.21 0.21
106 106
0.21
106
110
5.3
0.027 0.009
4.8
0.9906
calc. cone. RR LOQ LOD SD (N-1) CV
calculated concentration, accumulation factor taken into account Recovery rate related to the nominal concentration of the test item Limit of quantification Limit of detection standard deviation coefficient of variation
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1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro-
Alga, Growth Inhibition Test with Scenedesmus subspicatus, 72 h, Limit Test Project-No.020109CK
with the Water Accommodated Fraction (WAF), acc. OECD 201_____________ Study-No. SSQ84112
10.3 Results
Tab. 10: C o n cen tratio ns o f the A ctive In gredient 1-D ecanol, 3 ,3 ,4,4 ,5,5,6,6,7 ,7,8,8,9,9 ,10, 10,10-hep tad ecafluo ro -
Sampling Date
Start of Analysis
Test Item
Replicate No.
WAF
1 2
Control
LO Q [mg/L] LOD [mg/L] linearity ( r2 )
2002-04-01 Test Start
2002-04-05
2002-04-04 Test End
2002-04-05
1-Decanol, 3 ,3 ,4 ,4 ,5 ,5 ,6 ,6 ,7 ,7 ,8 ,8 ,9 ,9 ,1 0 ,1 0 ,1 0 -
heptadecafluoro-
Calc. Cone. [mg/L]
Calc. Cone. [mg/L]
0.56
< LOD
0.56
< LOD
0.39 0.37 < LOD < LOD
< LOD
< LOD
< LOD < LOD 0.027 0.009 0.9996
Mean'1 [mg/L]
0.47
calc. cone. WAF LOQ LOD
1)
calculated concentration, accumulation factor taken into account water accommodated fraction Limit of quantitation Limit of detection concentrations from the test end were not taken into account for mean value, because they ware < LOD
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1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro-
Alga, Growth Inhibition Test with Scenedesmus subspicatus, 72 h, Limit Test Project-No.020109CK
with the Water Accommodated Fraction (WAF), acc. OECD 201_____________ Study-No. SSQ84112
10.4 Calibration Curves
T h e calibration cu rves show n w ere estab lish ed for differen t concentration rang es and at differen t tim es
Calibration Curve of Fluowet (2002-04-04)
Fig. 2:
Calibration Curve o f Standard - concentration range 0.01 - 0.06 mg/L (dated 20 02-0 4-0 4)
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1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,1O-heptadecafluoro-
Alga, Growth Inhibition Test with Scenedesmus subspicatus, 72 h, Limit Test Project-No.020109CK
with the Water Accommodated Fraction (WAF), acc. OECD 201_____________ Study-No. SSQ84112
Calibration Curve of Fluowet (2002-04-10)
500000 450000 400000 ___ 350000 300000 250000
200000
<t 150000 100000 50000
0
0 2 4 6 8 10 Cone. [mg/L]
Fig. 3:
Calibration Curve of Standard - concentration range 0.1 10 mg/L (dated 2002-04-10)
Calibration Curve of Fluowet (2002-06-19)
Fig. 4:
Calibration Curve of Standard concentration range 0.1 - 0.6 mg/L (dated 2002-06-19)
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1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro-
Alga, Growth Inhibition Test with Scenedesmus subspicatus, 72 h, Limit Test Project-No.020109CK
with the Water Accommodated Fraction (WAF), acc. OECD 201_____________ Study-No. SSQ84112
The first calibration curve was established 2002-04-04 but the concentration range (0.01 - 0.06 mg/L) was not broad enough to be used for all tests running. Therefore the second curve (2002-04-10) which covers a much wider range (0.1 -1 0 mg/L) was used for the calculations in this test. The concentration calculations using the two different curves do not differ more than 10 % from each other. The calibration curve from 2002-06-19 with a concentration range of 0.1 - 0.6 mg/L shows that the GC/MS system has a good stability over a long period of time.
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Report 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoroAlga, Growth Inhibition Test with Scenedesmus subspicatus, 72 h, Limit Test with the Water Accommodated Fraction (WAF), acc. OECD 201__________
Page 33 of 39
Project-No.020109CK Study-No. SSQ84112
10.5 Chromatograms
Target Compound Report far #1 I w n 411 0.1 M .S M 8
S a m p le ID; Instrument D : Me m u lam ent Type; Acquisition D ate: Calculation Date: Sam ple Type: Inj. S a m p le N o te s : Divisor:
8411 0.1 I
Area 10.04.02 1 3:21 11.04.02 08:08 Calibration
1 ,0 0 0 0 0 0
Operator: L ast Calibration: Calteration Type: D ata File: M ethod;
11 .0 4 .0 2 0 6 :0 6 External Standard
c \ ... 8411 0.1 I j l .e m c\...\6 4 1 1 _rap 2 .m th
Multiplier:
1,000000
Param eter Compound Number Retention Tim e RT O ffset Peak Name C A S Number Status Q u an Ions Am a
H aig ht Am ount (RF) Fit Calibration Equationi B atalina C o d a
S p e c ific a tio n 4.918 /-0,200
69+395+463 >10
>=700 Linear, Include, N one
A ctu a l 1 4.911 m in. A 0,00676 m in. F tuowet 676-39-7
4368 2263 4 4 1 2 4 ,9 4 1 971 y * +4,4125e+4x +0 .0 BB
Statoa Pass
Paaa Pass
Fig. 5:
Chromatogram of Standard 0.1 mg/L (dated 20Q 2-04-10)
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1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafiuoro-
Alga, Growth Inhibition Test with Scenedesmus subspicatus, 72 h, Limit Test Project-No.020109CK
with the Water Accommodated Fraction (WAF), acc. OECD 201_____________ Study-No. SSQ84112
TargetCompound fe p a t to 1 from SS064112E WAF M-SMS
S a m p le ID: Iretium ant D : M e a c u t e m e rtf T y p e : Acquitftion Date: Calculation Date: Sam ple Type: Inj. S a m p le N o te s: D iviso r:
SS084112 E WAF I
Area 05 .0 4.02 23:49 0 6 0 4 .0 2 08:39 A nalyw a
1 ,0 0 0 0 0 0
Param eter Com pound Number Retention T im e R T Offset Peak Name C A S Number Status Quart lore
Area H eig ht Am ount Fit Calibration Equation Baseline C od e
Specification 4 ,9 2 5 /- 0 .2 0 0
69+395+463 >*10
>* 0,000
>=700 Linear, Ignore, None
Sudua and Effort:
M : Miaaing paafc.
Operator L ast Calibration: Calibraiion Type:
Data File: M ethod
05.04.02 13:46 External Standard c \ ... aao84112ew ef c\...\8 4 1 1 _rep 2 .m th
Multiplier.
1 .0 0 0 0 0 0
Afil
1 4,923 m in. 0 .0 0 1 8 3 min. Flu ow et 876-39-7
M
0 0
0,000 613 y * +2,0285e+4x +830.0136
Siati
Pass
Fig. 6:
Chromatogram of Test Item W a te r accom m odated fraction at the test end (dated 2 0 0 2 -0 4 -0 5 )
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Report 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoroAlga, Growth Inhibition Test with Scenedesmus subspicatus, 72 h, Limit Test with the Water Accommodated Fraction (WAF), acc. OECD 201__________
Page 35 of 39
Project-No.020109CK Study-No. SSQ84112
TargM Compound Import far f 1 fnm SSOB4111_2 E Ktr I.2.SM S
S a m p le ID: Instrum ent ID: Measurement Type:
Acquisition Oate: Calculation Date: Sam ple Type: Inj. S a m p le N o te s: D iviso r:
S S O M 1 1 1 JZ E Ktr 1
Area 0 5 .0 4.022 245 08.04.02 o & a o A n a ly a is
1X100000
Param eter Compound Number Retention Tim e R T Offset Peak Name C A S Number Status Q u an tons Area H eig h t Amount Fit Calibration Equation
Basatine C o d
S p e c ific a tio n 4 ,0 2 5 /- 0 ,2 0 0
60+385*463 >=10 >= 0,000 >=700 lin ear, Ignore, None
SUuaandEirara:
M : M W ng p*.
O p erato r L ast Calibration: Calibraikm Type:
D a ta F He: M ethod:
05.04.02 13:46 External Standard c \ ... a a o 6 4 1 11 2 e W c\...\8 4 1 1 _rep 2 .m th
M ultiplier Actual
1,000000
4,926 m in. A o ,00 M S min. Flu o w et 678-3-7
M
0
0 0 ,0 0 0 465 y * +2Q265e+4x +630,0136
S ta to i Pass
Fail Pass Fail
Fig. 7:
Chromatogram of Control at test end (dated 20 0 2 -0 4 -0 5 )
000078
Dr.U.Noack-Laboratorium fr Angewandte Biologie
Report 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro-
Page 36 of 39
Alga, Growth Inhibition Test with Scenedesmus subspicatus, 72 h, Limit Test Project-No.020109CK with the Water Accommodated Fraction (WAF), acc. OECD 201_____________ Study-No. SSQ84112
11 Raw Data
F lu o re s c e n c e a t te s t b e g in n in g
control
cuvette no.
F1
F2
F3
F4
F5
1
91 90
92
94
94
2
88 87
87
92
94
3
92 65
88
90
97
4
76 73
78
83
77
5
85 85
87
90
88
6
80 82
83
82
82
i
E
con e.:
W A F (0 .47
cuvette no.
F1
F2
F3
F4
F5
1
92 96
94
5
95
2
87 89
86
89
88
3
86 84
84
87
98
SB: Ku
P rO f-N r. P ro je k t-N r.
020109CK SS084112
F6
m v F3-F6
eorr. fluor.
91 9 2 .7 5
9 2 .7 5
90 90.75
9 0 .7 5
91 9 1 ,5 78 79
9 1 ,5 79
87 88
88
85 83
83
F6
mv F3-F6
corr. fluor.
98 95 ,5
95 ,5
92 89.25
8 9 ,2 5
89 89,5
89 ,5
VC 0 ,0 1 6 2 0 ,0 3 2 9 0 ,0 4 2 3 0 ,0 3 4 3 0 ,0 1 6 1 0 ,0 1 7 0
overall m v 87 ,5
VC(% ] 0 ,0 1 8 1 0 ,0 2 1 2 0 ,0 6 7 3
overall m v 91 ,41866667
F lu o re s c e n c e a fte r 2 4 h
SB : Ku
PrQf-Nr. P ro je k t-N r.
020109CK SS064112
control
cuvette no.
F1
F2
F3
F4
F5
F6
m vF3-F6
corr. fluor.
1
387 375
388
381
3B9 405
3 9 0 ,7 5
39 0.75
2
459 442
457
441
436
453
4 4 6 ,7 5
4 4 6 ,7 5
3
481 471
475
474
466
462
4 6 9 ,7 5
46 9,75
4
460 470
467
448
462
456
4 5 8 ,2 5
45 8,25
5
451 471
461
464
460
474
6
474 472
465
455
465
472
4 6 4 ,7 5 4 6 4 ,2 5
46 4,75 4 6 4 ,2 5
cone.: WAF (047 mgft.)
cuvette no.
F1
F2
F3
F4
F5
F6
m v F3-F6
corr, fluor.
1
561 567
562
564
571
573
5 6 7 ,5
5 6 7 .5
2
491 472
462
462
463
457
461
461
3
411 418
432
428
437
423
430
430
V C [%] 0 ,0 2 6 0 0 ,0 2 2 1 0 ,0 1 2 8 0 ,0 1 7 9 0,0138 0,0151
overall m v 4 4 9 ,0 8 3 3 3 3 3
V C [%1 0 ,0 0 9 4 0 ,0 0 5 9 0 ,0 1 3 8
overall m v 4 8 6 ,1 6 6 6 6 6 7
000079
Dr.U.Noack-Laboratorium fr Angewandte Biologie
Report
Page 37 of 39
1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro-
Alga, Growth Inhibition Test with Scenedesmus subspicatus, 72 h, Limit Test Project-No.020109CK
with the Water Accommodated Fraction (WAF), acc. OECD 201_____________ Study-No. SSQ84112
F lu o re s c e n c e a fte r 4 8 h
c ovatta no. 1 2 3 4 5 6
c o n tro l F1
2668 2782 2672 2676 2942 2856
F2 2598 2834 2788 2720 2996 2824
F3 2602 2822 2768 2732 2986 2836
cone.:
cuvette no. 1 2 3
W AF (0.47 m p/U
F1 F2 2684 2734 3254 3260 2792 2824
F3 2814 3212 2860
F4 2592 2826 2768 2684 2940 2612
F4 2790 3254 2840
F5 2546 2804 2756 2686 2924 2854
F5 2758 3236 2880
SB: Ku
F6 2540 2786 2732 2664 2962 2824
m v F3-F6 2570 2 8 0 9 ,5 2756 2 6 9 1 .5 2953 2 8 3 1 ,5
F6 2760 3226 2984
m vF3-F6 2 7 8 0 ,5 3232 2891
PrOf-Nr. Projekl-Nr.
020109CK SSO B4112
corr, fluor. 2570
2 8 0 9 ,5 2756 2 6 9 1 ,5 2953 2 8 3 1 ,5
VC[% ] 0 ,0 1 2 3 0 ,0 0 6 5 0 ,0 0 6 2 0 .0 1 0 7 0,0091 0 ,0 0 6 3
overall m v 2 7 6 8 ,5 8 3 3 3 3
corr. fluor. 2 7 8 0 ,5 3232 2891
V C |% ] 0 ,0 0 9 6 0 ,0 0 5 5 0 ,0 2 2 2
overall m v 2 9 6 7 ,8 3 3 3 3 3
F lu o re s c e n c e a fte r 7 2 h
cuvette no. 1 2 3 4 5 6
control F1
9140 9770 9690 9800 9060 9730
F2 9040 9720 9760 10030 9120 10120
F3 8620 9610 9450 10170 9250 10140
co n e.: cuvette no.
1 2 3
W A F (<M 7 m gfl.) F1 F2
9310 9520 10240 10180 10210 10130
F3 9590 10080 10010
F4 8900 9430 9510 10090 9170 10080
F4 9270 9910 9920
F5 8630 9410 9330 9940 9060 10170
F5 9300 9B20 9550
SB: Ku
F6 8690 9300 9260 9650 8950 9850
m v F3-F6 8760 9 4 3 7 ,5 9 3 8 7 ,5 9 9 6 2 .5 9 1 0 7 ,5 10060
F6 9210 9650 9560
m v F3-F6 9 3 4 2 ,5 9665 9760
Pr0f-Nr. Prcjekt-Nr.
0 20 109CK S S 0 8 4 112
corr. fluor. 8780 9 4 3 7 ,5 9 3 8 7 ,5 9 9 6 2 ,5 9 1 0 7 ,5 10060
V C |% ] 0 ,0 1 4 0 0 ,0 1 3 6 0 ,0 1 2 1 0 ,0 2 3 0 0 ,0 1 4 4 0 ,0 1 4 4
overall m v 9 4 5 2 ,5
corr. fluor. 9 3 4 2 ,5 9865 9760
V C [%] 0.0181 0 .0 1 8 2 0 ,0 2 4 5
overall m v 9 6 5 5 ,8 3 3 3 3 3
000080
Dr.U.Noack-Laboratorium for Angewandte Biologie
Report
Page 38 of 39
1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro-
Alga, Growth Inhibition Test with Scenedosmus subspicatus, 72 h, Limit Test Project-No.020109CK
with the Water Accommodated Fraction (WAF), acc. OECD 201_____________ Study-No. SS084112
12 Certificate of 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10heptadecafluoro- Analysis
CUriaai G m b H , Wrtt Gandorf
CQG ! Qm IM u h u m c m m ih
D-MSO* BurtUichM Fu: 4-4-49 4679 / 1676
C la ria n t
1
Inspection
certificate
according to EN10204-3-1B
Date: 13.11.2001 Page: l / l
Oar consignment Material
Material-no. No.
1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10, 1C-Keptadecflfluoro : 104705 : POO/OD1
wti z - t b a t e .', o i w n l.:r r.it-# r o n s ig m - e n t i s a p a r t , th e f o l l o w i n g '/ a le e *
-ire cf-.th rm in cc: : i `. e y c o n :a im i:o t.va a g r e e d p r o d u c t s p e c i f i c a t i o n
Ir.s c e c t i
c la r a v :: ; x z \'i> - m e c h o c s p e c i f i c a t i o r i
R e s u lt
Z 3 -? e r f* u o r a *k,1 s t a r.c 1 ..'apl.nry ,C
urn Periiuocoa l k y l e t h a n o l Capillary GC
Sum Partluoralkylbutanol Capillary GC
Sum ?er:luorsikylethene Capillary GC
Water content Kari Fischer DIN 51777
N-Merhylcyrrolidone content Capillary GC
A 1 O
>- 97,5 >- 98,0
<0, 5
<-0, 2
o V
<N O V
9 9 , 3 \ [a/a! 99,9%[a/a] < 0, U [ a / a i < 0 , 1 % [a/a]
< 0, li
The above particulars co not release the customer from the obligation to carry cut an inspection z'i goods received.
>!r . Saumg-irtr.ar - Wcrksir.spector
OOOOSl
Dr.u .Noack-Laboratorium for Angewandte Biologie
Report
Page 39 of 39
1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro-
Alga, Growth Inhibition Test with Scenedesmus subspicatus, 72 h, Limit Test Project-No.020109CK
with the Water Accommodated Fraction (WAF), acc. OECD 201_____________ Study-No. SSQ84112
13 GLP-Certificate of Dr.U.Noack-Laboratorium
Nisdsrsechsisches Umvmttmlnirtarium
Guta Laborpraxis/Good Laboratory Practice
G LP-Bescheinlgung/Statem ent o f GLP Com pliance
(gem M Vaccording to 1 B b A b s. 1 C h a m ikalle n g e setz)
E in e G L P -ln sp e k tlo n zu r berw achung d a r E in h altu n g d er G L P -G ru n d s tfz e gem tft C h e m lka llon g esetz bzw . R ich tlin ie 8 8 /3 20 /E G w urde durchgefQ hrt In:
A sse ssm e n t o f conform ity w ith G L P acco rding to C h e m k slie n g e se tr and D irective 8 8 /3 2 0 /E E C at:
X P r fe in ric h tu n g / T e s t fa d llty
O P r fsta n d o rt / T e s t site
D r. U . N O A C K L A B O R A T O R IU M f r a n g e w a n d te B io lo g ie K a th e -P a u lu s -S tr. 1 D -3 1 15 7 Sarsted t
D r. U . N O A C K L A B O R A T O R Y o f a p p lie d b io lo g y K a th e -P a u lu s -S tr. 1 D -3 1 1 5 7 Sarsted t
(Unvsrw ocS M IS sr B w le tin u n g und a dW M AIlw sutw ocat n a n e a n d irlrVs -- }
P r fu n g e n n a c h K a te g o rie rV A re a s o f E x p e rtis e
(OwnSS/soconUng C te n A M V -O L P Nr. 5.3 /O eC O guldsnce)
1 - P r fu n g e n z u r B e stim m u n g d e r p h y s ik a lis c h -c h e m is c h e n E ig e n s c h a fte n u n d G e h a ttsb e s tlm m iin g e n 3 - P r fu n g e n z u r B e stim m u n g d e r e rb g u tv e rO n d e m d e n E ig e n s c h a fte n (in v itro u n d in v iv o ) 4 - Umwelttoxikologische Prfungen zu Auswirkungen auf aquatlscha und terrestrische Organismen 5 - P r fu n g e n zu m V e rh a lte n Im B o d e n , kn W a s s e r u n d in d e r L u ft. P r fu n g e n z u r B io a k k u m u la tio n u n d
z u r Metabollslerung 6 - P r fu n g e n z u r B e stim m u n g v o n R c k st n d e n 7 - P r fu n g e n z u r B e stim m u n g d e r A u sw irk u n g e n a u f M e so k o sm e n u n d n a t rlic h e k o s y s te m e
D atu m d e r In sp e k tio n / D a te o f In sp e ctio n fTse-MonaUshr / dsy.monlh.year)
25 . - 27. Ju n i 2001 / Ju n e 25*' - 27* . 2001
2 9 . A u g u s t u n d 2 1 . S e p te m b e r 2 0 0 1 / A u g u s t 2 9 * 'a n d S e p te m b e r 2 1 " , 2 0 0 1
D ie g e n a n n te PrDfelnrtchtung/P a r g e n a n n te OrtHrta n dart
befind et
sich
im
n ation alen
QLF-
O b e r w a c h u n g s v e r fa h r e n u n d vrtrd re g e lm a ftig a u f
E in h altu n g d a r G L P -G r u n d s ttx e Oberw ach t.
A u f d e r G ru n d la g e d e s In sp ektio n sberich lea w ird hierm it b e s t t ig t , d a s s in d i e s e r P r O fe ln r ic h tu n g M Ie s a m PriWata n d e rl d is o b e n g e n a n n te n P r fu n g e n u n te r E inhaltung d e r G L P -G ru n d sA tze du rchgefbhrt w erden knnen.
T h e a b o v e m e n tio n e d te a t fa cility / te s t s ite la In clu d e d in th e n a tio n a l G L P C o m p lia n ce P ro g ra m m e a n d la In sp ected on e regular b a sis.
B a a e d o n t h e in s p e c tio n re p o rt It c a n b e c o n firm e d , th a t th is te at facility / te st site la a b le to co n d u ct th e a fo re m e n tio n e d stu d ie s in co m p lia n ce w ith th e P rin cip le s of GLP.
1 1 . M flrz 20 0 2
Im A u ftr g e
y
Dr. B raedt
000082
