Document JXMR7rvkmBaEXVz9rebJER0e
BIODEGRADATION
TEST SUBSTANCE
Identity:[2-(N-Ethylperfluorooctanesulfonameitdhoy)lacrylatem;ay alsobe referredtoas Bl 228, D-1, ETFOSEA, or FX-13.(2-Propenoicacid,2[ethyl[heptadecafluorooctyl)sulfonyl]aminoe]settehry,ClAS # 423-825)
Remarks: Materialisan amber solid.Sample puritynot recorded.3M lot number 13.
METHOD
Method: Not noted. Type: Aerobic GLP: No Year completed: 1984 Test solutionpreparation:Sample weighed onto a coverglass,dissolvedin acetone and washed intoa bottlewithacetone. Bottlecoated withacetone solution.Acetone was then purged from the bottlewithnitrogen. Contact time:20 days Inoculum: Not noted
RESULTS
Test substance: 5-day BOD: 10-day BOD: 20-day BOD:
COD:
Not measurable 18,990 mg/kg 23,000mg/kg 240,000 mg/kg
Reference substance: The glucose-glutamiaccid standardachieveda BOD of 201 mg/L in5-days,which iswithinthe recommended range forthe Standard Methods BOD testmethod.
CONCLUSIONS
This testingindicatedthatthe testsubstance isnot capable ofreadily biodegradinginaerobicenvironments.
DATA QUALITY
ReliabilitKyl:imiscrhankin3g.Thisstudyw,hilewellconductedusingthe methodology acceptableatthe time,lacksanalyticadlata fordeterminationof the sample purity.Testingwas alsodone wellabove the solubilitlyimiotfthe test substance withthe use ofacetone as a solubiliziangent.
REFERENCES This study was conducted by the3M Company, EnvironmentalLaboratory,Lab Request number B1228, 4/26/84
OTHER Submifter: 3M Company, EnvironmentalLaboratory,P.O. Box 33331, St.Paul, Minnesota,55133
Last changed: 5/22/00
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BIODEGRADATION (ModifiedMITI Test)
TEST SUBSTANCE
Identity:[2-(N-Ethylperfluorooctanesulfonamiedtoh)ylacrylate;may also be referredtoas Bl 228, D-1, ETFOSEA, or FX-13.(2-Propenoicacid,2[ethy[lheptadecafluorooctyl)sulfonyl]amino]eesttheyrl,CAS # 423-825)
Remarks: Materiiaslan ambersolidR.eportlistssampleas> 99%,as provided by sponsor.The pudty/identiotfythetestsubstance cannot be substantiated.As presentedinthe report,thestructuraflormula indicatesthat"purityc"annot be assigned as "99% or more". Lot number 4.
METHOD
Method: OECD 301C Type: Aerobic GLP: Yes Year completed: 1995 lnoculum source: Chemical BiotestinCgenter,Chemical Inspectioannd TestingInstitutJea,pan
Remarks: Itappears thatthe inoculumwas obtainedfrom onlyone source, ratherthan theten sources requiredinthe method.
RESULTS
Test substance: After28-days,the percentbiodegradabilitoyfthe test substance was determinedby threemethods: BiochemicalOxygen Demand (BOD), DissolvedOrganicCarbon (DOC), and ResidualTest Substance Concentration.The resultsareas follows:
Percent Biodegradability(of3 replicates): BOD: 4,-1,and 6% DOC: Not calculatedt;estsubstance not dissolvedinsolution Test Substance Concentration:5 and 1% (onlytwo determined)
Reference substance: The anilineachieved68% biodegradabilitiyn28-days.
Measurement of testsubstance: The testsubstance concentrationwas only measured atthe end ofthestudy. Although thislaboratorfyollowedthe
procedures as outlinedinOECD 301 C, the startincgoncentrationshould also be determined ratherthan relyingon nominalconcentrations.Inaddition, subsamples were removed from the bioreactorfsorpH and DOC determinations atthe end ofthe study. The sample used forpH determinationsand the remainingsample aftercentrifugatifoonrDOC were returnedtothe bioreactor priortosamplingforthe measurement ofthetestsubstance concentration. These samples should have been removed afterthe bioreactorswere sampled fortestsubstance concentrationdetermination.
Calculationsoftheoreticaolxygen demand should have made which account for possibleoxygen consumption due to nitrification.
CONCLUSIONS
This testingindicatedthatthe testsubstance isnot capable ofreadily biodegradinginaerobicenvironments.
DATA QUALITY
ReliabilityK:limischranking3.There isno referenceto thesolubilitiynformation providedinthe study. The quoted purityfidentoifttyhe testsubstance isnot substantiated.Measurement oftestsubstance concentrationwas only performed atthe end ofthe study.The bioreactorwsere potentially contaminated by the practiceofreturningsubsamples afterpH and DOC determinations.The calculatioonftheoreticaolxygen demand should have been perfon-nedusingthe correctionfornitdfication.
REFERENCES Study conducted atthe requestof Sumitomo 3M LTD by MitsubishiChemical Safety InstitutLetd.,Yokohama, Japan
OTHER
Submifter: 3M Company, EnvironmentalLaboratory,P.O. Box 33331, St.Paul, Minnesota, 55133
Last changed: 5/22/00
M.S.I. REPORT No.lB363(2)G
Ready Biodegradability Test of D-1 Submitted to
SUMITOMO 3M LIMITED
Prepared by Mitsubishi Chemical Safety Institute Ltd.
October 31, 1995
Ready Biodegradability Test of D-1 (English version)
Study No. Study Title Sponsor
: lB363(2)G : Ready Biodegradability Test of D-1 : SUMITOMO 3M LIMITED
This report was conducted in the Yokohama Laboratory of the Mitsubishi Chemical Safety Institute Ltd., 1600 Kamoshida-cho, Aoba-ku, Yokohama 227, Japan.
This report is an English version of the orginal, which was written in Japanese. The underesigned hereby declare that this version faithfully reflects the original report to the best of our knowledge.
Mitsubishi Chemical Safety Institute.Ltd.
Date Kikuo Yoshida, Ph.D. Environmental Science Division of Yokohama Laboratory
Approved by
. Jun Toriya, B si@@ Head of Yokohama Laboratory
Date
COMPLIANCE WITH THE GLP STANDARDS
Study No. Study Title Sponsor
: IB363(2)G : Ready Biodegradability Test of D-1 : SUMITOMO 3M LIMITED
To the best of our knowledge and belief, the study described in this report was conducted in compliance with the Good Laboratory Practice (G.LP)standards applied to industrial chemicals under the Chemical Substances Control Law.
Facility management Sakae Kpike, B.Sc. Facility management Head of Yokohama Laboratory
Sealed date :April 28, 1992
Study director Kikuo Yoshida, Ph.D. Study Director Environmental Science Division
Sealed date :April 28, 1992
QUALITY ASSURANCE STATEBENT
Study No. Study Title Sponsor
: lB363(2)G : Ready Biodegradability Test of D-1 : SUMITOMO 3M LIMITED
We hereby. certify that the above study has been performed in accordance with the Good Laboratory Practice (GLP) standards applied to industrial chemicals under the Chemical Substances Control Law.
Dates of inspections and reportings to the Study Director and the Facility Management are as follows,:
In-progress study Final report
Date of Inspection
March 18, 1992 April 15, 1992 April 16, 1992
April 28, 1992
Date of Reporting_
March 30, 1992 April 15, 1992 April 16, 1992
April 28, 1992
Quality Assurance Staff Junko Katoh, Ph.D.
Yumiko Miura, B.Sc.
Sealed date :April 28, 1992 Sealed date :April 28, 1992
We, the undersigned declare that the work was performed according to the procedures herein described, and that this report provides a true and correct record of the results obtained.
Experiment Staff
Ayumi Okuda, M.Sc.
Sealed date :April 28, 1992
Environmental Science Division
Junko Koyasu, B.Sc. Environmental Science
Division
Sealed date
:April 28, 1992
Yukiko Mizoguchi
Sealed date :April 28, 1992
Environmental Science Division
Staff for cultivation of activated sludge
Yasuo Sato
Sealed date :April 28, 1992
Environmental Science Division
CONTENTS
Page
EXPLANATORY
REMARKS ...............................................- .7
SUMMARY ...........................................................8...
1 STUDY TITLE .......................................................9...
2 PURPOSE ..........................................................9...
3 TEST GUIDELINES ...................................................9...
4 GLP COMPLIANCE ..............................................
9
5 STUDY PERIOD ..............................................
9
9 6 SPONSOR ..............................................*-.,.
9 7 TESTING FACILITY .........................................
8 STORAGE AND RETENTION OF TEST SUBSTANCE AND RECORDS .................10
9 TEST SUBSTANCE ....................................................11..
10 CONFIRMATION
OF IDENTITY AND STABILITY .............................1.2
10.1 . IDENTITY ...........................................*.**.**** 12
10.2
STABILITY ...........................................*.-.-*** 12
11 TEST 11.1 11.2 11.3 11.4 11.5 11.6 11.7
13 METHODS ..........................................
ACTIVATED
SLUDGE ....................................
13
EXPOSURE CONDITIONS
.....................................I.-.13
14
BOD MEASUREMENT
....................................
14
pH MEASUREMENT
....................................
DOC MEASUREMENT
...................................
14
MEASUREMENT
OF RESIDUAL TEST SUBSTANCE CONCENTRATION
........15
EQUATIONS FOR CALCULATION
OF DEGRADABILITY
..................1.6
17 12 RESULTS ...............................................
12.1 12.2 12.3 12.4 12.5
OBSERV4TION pH MEASUREMENT ACTIVITY OF DEGRADABILITY DEGRADABILITY
OF TEST BOTTLES AFTER EXPOSURE PERIOD ...........17
................................*.*****'****** 17
SLUDGE ................................*.***,*,'' 17
BASED BASED
ON BOD .................................1.7 ON DOC .................................17.
12.6
DEGRADABILITY TEST SUBSTANCE
BASED ON THE RESIDUAL
...... 18
CONCENTRATION
...........................
13 CONCLUSIONS TABLES AND
18 ...................................*.-.,**,******'*''*''*
FIGURES
19-33 ................................................
EXPLANATORY
REMARKS
Study No. Study Title
lB363(2)G Ready Biodegradability
Test of D-1
The ready biodegradability test of D-1 was originally started on January 29, 1992, and the BOD measurement of the test'substance was initiated on February 13, 1992 (Study No. IB363G) . However, the degradability value of aniline based on the BOD measurements did not exceed 40%.
Therefore, the test was terminated on February 20, 1992 and the new test (Study No. lB363(2)G) was started with the same test substance and the same test method after approval of the sponsor.
Some of data used in this report (IR chart, calibration curve and recovery) are those obtained in the former study (Study No. lB363G).
SU14MARY
STUDY TITLE
Ready Biodegradability
Test of D-1
METHOD The method described in the Order, 3ahich prescribes the procedure of
testing new chemical substances as required by the Chemical Substances Control Law(Japanese Law 117, 1973). ("Ready biodegradability, Modified MITI test" in OECD Guidelines for Testing of Chemicals).
Test bottle Measurements
Bottle 1 Bot tle 2
Bott* les 3-5 Bottle 6
: aniline + sludge + basal medium : sludge + basal medium : test substance + sludge + basal medium : test substance 4 water
Biochemical oxygen demand (BOD) Dissolved organic carbon (DOC) Residual test substance concentration
(for 28 days) (at day 28) (at day 28)
RESULTS
Measured values (day 28)
BOD, DOC, Test value
mg mg/l substance, mg/l corrected with BOD
Bottle No.
3
4
5
6
1.0* -0.3* 1.6* 0.0
0.7* 0.3* 1.6* 2.0 101.3 105.2 91.5 106.7 or DOc value of Bottle 2.
Theoretical value 27.7 28.8 100.0
Degradabilities Bottle No.
3
4
5
BOD DOC Test
substance
4
-1
6
NA*
NA*
NA*
5
1
NA**
not calculated because the test substance was not dissolved in the test
solution. not calculated because the test substance might be lost accidentally during
condensation of the solution under vacuum.
CONCLUSION From the degradability results based on the BOD and the residual test
substance concentration , it can be concluded that the test substance is not readily biodegradable and is not transformed under the conditions prescribed by the Modified MITI test.
1
STUDY TITLE
Ready Biodegradability
Test of D-1
2,
PURPOSE
This study was conducted
substance for notification
to evaluate
the ready biodegradability
of the test
under the Chemical Substance Control Law of Japan.
3
TEST GUIDELINES
The test was conducted
according
to the method
described
in the Order,
which prescribes the procedure of testing new chemical substances as required
by the Chemical Substances Control Law (Japanese Law 117, 1973). This method
is also known as the "Ready biodegradability, Modified MITI test" in the OECD
Guidelines for the Testing of Chemicals No. 301C.
4
GLP CM4PLIANCE
The test was conducted
according to the Good Laboratory
Practice
(GLP)
standards applied to industrial chemicals under the Chemical Substances
Control Law.
5
STUDY PERIOD
March 17, 1992 to April 28, 1992
(BOD measurement : March 18, 1992 to April
15, 1992)
6
SPONSOR
SUMITOMO 3M LIMITED
8-8 Minamihashimoto
3-chome, Sagamihara 229, Japan
7
TESTING
FACILITY
Yokohama
Laboratory
Mitsubishi Chemical
Safety Institute
Ltd.
(M.S.I.)
1000 Kamoshida-cho, Aoba-ku, Yokohama 227, Japan
Head Office: Mitsubishi Chemical Safety Institute Ltd. (M.S.I.)
1-30 Shiba 2-chome, Minato-ku, Tokyo 105, Japan
8
STORAGE AND RETENTION OF TEST SUBSTANCE AND RECORDS
The following records, the raw data, and a small sample of the test
substance will be stored in the archives of M.S.I. for 10 years after the
submission of the final report. After this period, the sponsor will be
contacted for the approval of the disposal of data. Data can be stored in the archives for a further specified period at the sponsor's request with an
additional fee.
1) Protocol.
2) Final report.
3) Raw data.
4) Quality assurance reports. 5) Test substance (approx. 2 g). 6) other documents required under
the GLP standards.
9
TEST
1)Name*
SUBSTANCE
2)Chemical name*
:D-1 (M.S.I. identification No. lB363(2)G)
:2-(N-Ethyl-N-perfuloroalkyl(C=1-8)sulfonylaminolethylacrylate
3)Structural formula*
C2H5
0
-1
11
Cn F2n+l@02-N-CH2CH20-C-CH=CH2
4)Elemental composition
(n.-@::1-8,main component; n=B)
:C;28.8%, H; 1.9%, N; 2.3%, 0; 9.8%, F;52.0% S; 5.2% (Measured by M.S.I.)
5)Physico-chemical Solubility*
properties
insoluble in water insoluble in DMSO >50% in acetone
Melting point* Boiling point* B)Batch* 9)Purity* 10)Appearance ll)Date of receipt 12)Supplied quantity*
:27-420C
:ca. 150*C/lmmHg
:Lot No. 4 :>99% (n=1-7;ca.21
%(total), n=8;ca.78%)
:amber solid
:September 25, 1991
:100 g
provided by the sponsor.
10
CONrlRb&kTION OF IDENTITY AND STABILITY
10.1 IDENTITY For the identification of the test substance, the infrared absorption
spectrum of the test substance was measured. The measured spectrum was consistent with that supplied by the sponsor.
1)Apparatus
: Perkin-Elmer
model
1640 infrared
spectrophotometer. [Figure I (P-23)]
10. 2 STABILITY After the exposure period, the infrared absorption spectrum of the test
substance that had been stored in a refrigerator during the course of the study was measured. The spectrum was consistent with that shown in Fig. 1, indicating that the test substance was stable under the storage condition.
I)Apparatus
Perkin-Elmer
model 1640 infrared
spectrophotometer. [Figure 2 (P-24)]
11
TEST METHODS
The test was performed
according to the method described
in the Order,
which prescribes the procedure of testing new chemical substances as required by the Chemical Substances Control Law (Japanese Law 117, 19'73).
This method is also known as the "Ready biodegradability, Modified MITI
test" in OECD Guidelines for Testing of Chemicals No.301C (1981).
The test substance was exposed to the activated sludge in a closed-system
oxygen consumption measuring apparatus. The biochemical oxygen demand (BOD)
was measured over a 28 day period. After this period, the concentrations of
the dissolved organic carbon (DOC) and the residual test substance in the
test bottles were measured. The biodegradability of
the
test
substance
was evaluated
from these
results.
11.1 ACTIVATED
SLUDGE
1)Mixed liquor suspended
solid (MLSS)
3300 mg/l
2)Source
Chemical Biotesting Center, Chemical Inspectibn and Testing'Institute, Japan
3)Date of receipt.: January 23, 1992
11.2 EXPOSURE
CONDITIONS
1)Temperature
25 1 *C
2)Exposure period
28 days
3)Test volume 4)Concentration
300 ml
test substance (bottles 3-6) aniline*' (bottle 1) activated sludge (bottles 1-5)
: 100 mg/i : 100 mg/i
30 mg/l
Reference substance : Syowa Chemicals, Lot No.SC-2726
5)Test bottle Bottle 1
contents :
: Aniline + activated sludge + basal medium
29Rl(30 mg) of aniline was added to the basal
mediUM*2,
then activated sludge was added.
Bottle 2
: Activated sludge + basal medium Activated sludge was added to the basal mediUM*2.
Bottles 3-5
: Test substance + activated sludge + basal medium 30 mg of the test substance was added to the basal
medium*2, then activated sludge was added.
Bottle 6
Test substance + deionized water 30 mg of the test substance was added to 300 ml of deionized water (purified by Milli-Q).
*2 The total volume of the basal medium and the sludge was held fixed at
300 ml.-
11.3 BOD MEASUREMENT
The BOD was measured for 28 days.
1)Apparatus
closed system oxygen consumption measuring apparatus Ohkura Electric Co., Model OM-2001 (M.S.I. ID No.B).
11.4 pH MEASUREMENT After the exposure period, 20 ml-of the test solution in each test bottle
was transferred into a 20 ml glass beaker for pH measurement. After pH measurement, the test solution was returned to each bottle for
measurement of residual test substance concentration.
1)Apparatus
pH meter, Denki Kagaku Keiki, Model
COM-10
11.5 DOC MEASUREMENT
The concentration of the DOC was measured as follows.
1)Apparatus
TOC analyzer, Shimadzu Co., model TOr--5000
2)Conditions
Furnace temperature Air flow rate Sensitivity Injection volume
: 680*C(TC) : 150 ml/min. :x 5 : 50 ILl
3)Calibration curve The following standard solutions were injected into the TOC analyzer.
The calibration curve was prepared by the data processor of the analyzer.
standard solutions
TC(total carbon)
20 and 50 mg C/1 aqueous solutions of potassium biphthalate.
IC(inorganic
carbon)
0 mg C/1 deionized water (purified by Milli-Q) and 10 mg C/1 aqueous solution of sodium hydrogen
carbonate.
4)Measurement of DOC in the bottles Ten ml of the test solution in each bottle was transferred into a 10 ml
centrifuge tube and centrifuged at 3000 rpm for 10 minutes. Five ml of
supernatant was used for the measurement of DOC. The remaining supernatant and the precipitate were returned to each
bottle for the measurement of the residual test substance concentration.
11.6 MEASUREMENT OF RESIDUAL TEST SUBSTANCE CONCENTRATION The concentration of the residual test substance was measured
by the gas
chromatography (GC) as follows:
1)Apparatus Gc Integrator
:Hewlett-Packard. Model 5890A (No.2) with FID detector :Hewlett-Packard. Model 3392A
2)Conditions Column
Temperature
Carrier gas Detector flow Attenuation Injection volume
:J & W. DB-17(50%Phenyl 50%methyl polysiloxane), 30 m x 0.25 mm i.d., 0.25@= (film thickness) (Column oven) 100*C (lmin.) -10OC/min. -200*C (lmin.)
(Injector]250*C, (Detector]280*C :He 30 ml/min. :H2 40ml/min., air 400 ml/min. :[GCI 1 V :1 Kl
3)Calibration curve The standard solutions of the test substance with 0, 250, 500, and 1000
mg/l in ethyl acetate were analyzed. The total peak area of test substance
was calculated by adding the area of the peaks with retention time of
about 5.6, 6.2 and 6.5 min. The total area was plotted against the
concentration. The correlation coefficient was calculated to be 1.00 by
the method described in Japanese Industrial Standards Z9041-1966. (Figure 3 (P-25) and Figure 4 (P-26)]
4)Recovery test Duplicate test bottles (identical to the test bottles 3-5 described in
11.2) were kept in a closed-system oxygen consumption measuring apparatus at 251*C for 30 minutes.
Each test solution was treated by the procedures described in 11.6.6. The concentration of the test substance was measured by GC. The average recovery of the test substance was 97%.
(Table 3 (P-21) and Figure 5 (P-27)] The concentration of the residual test substance measured for bottles 3 -6 were corrected with the average recovery value. 5)Detection limit The detection limit of the test substance was calculated to be 3 mg/l based on the minimum detectable peak area of 100000 @LV- sec for the largest peak (R.T. 6.2 min.) in the GC chromatogram.
(Table 3 (P-21) and Figure 5 (P-27)]
6)Measurement of test substance in test bottles The contents in each bottle was separately transferred into a 500 ml
glass separatory funnel, which was then extracted twice with 100 ml of
ethyl acetate. The extract was filtered through a grass funnel containing 50 g sodium sulfate, anhydrous to remove water. Then filtrate was condensed to ca. 20 ml under vacuum (<500C). The solution was brought up
to 50 ml with ethyl acetate. The ethyl acetate solution was then analyzed by GC. The concentration
of the residual test substance was determined as described in 11.6.2.
11.7 EQUATIONS FOR CALCULATION OF DEGRADABILITY Equations for calculation of degradability based on BOD, DOC, and
residual test substance concentration are as follows:
I)Degradability based on BOD
Degradability (%) = (BODS- BODb)/ThOD X 100
where
BODs BODB THOD
Oxygen consumption Oxygen consumption Theoretical oxygen test substance.
(mg) in bottles 1, 3, 4, and 5. (mg) in bottle 2. demand (mg) of aniline and the
THOD
aniline
: CGH7N THOD
+ 35/402 - 6CO2 + 7/2H20 : 90.2 Mg 02/30 mg-aniline
+ N02
test substance : C+02-CO2,
H+1/402-1/2H20,
N+02-NO2,
S+02-S02,
F-F
THOD : 27.7 Mg 02/30 mg-test substance
2)Degradability based on DOC
Degradability (%) = (1 - (DOCs-DOCb)/DOCc)XlOO
where
DOCs DOCB DoCc
DOC (mg/1) in bottles 3, 4, and 5. DOC (mg/1) in bottle 2. DOC (mg/1) in bottle 6.
3)Degradability based on residual test substance concentration
Degradability (%) = (1 - Cs/Cc) X 100
where Cs Cc
: concentration (mg/1) in bottles 3, 4, and S. : concentration (mg/1) in bottle 6.
12 RESULTS There was no specific
circumstances
bility of the test results.
which might have affected
the relia-
12.1 OBSERVATION OF TEST BOTTLES AFTER EXPOSURE PERIOD The solutions in the bottles were colorless except the bottle 1, in which
the solution was cloudy. Growth of the sludge was observed in bottle 1 in contrast with the bottle
2. No growth was observed in the bottles 3, 4 and S.
12.2 pH MEASUREMENT After 28 days of exposure, the pH was determined to be 7.4, 7.4, 7.5 and
7.4 for bottles 3, 4, 5 and 6, respectively.
[Table 1 (P-19))
12.3 ACTIVITY OF SLUDGE
The degradability value of aniline based on the BOD measurements was 59%
after 7 days. The activity of the sludge was thus shown to be satisfactory. (Table 1 (P-19) and Figure 6 (P-26-31)]
12.4 DEGRADABILITY BASED ON BOD BOD*L in bottles 3, 4, and 5 (as corrected with the value in bottle 2)
were 1.0, -0.3 and 1.6 mg, respectively. BOD in bottle 6 was 0.0 mg. (*' Maximum theoretical value = 27.7 mg) -
The degree of degradability based on the BOD measurements were 4, -1 and 6% for bottles 3, 4, and 5, respectively.
[Table 1 (P-19) and Figure 6 (P-28-31)]
12.5 DEGRADABILITY DOC*2 in bottles
BASED ON DOC 3, 4 and 5 (as corrected
with the value in bottle 2) were
0.7, 0.3, and 1.6 mg/l, respectively. DOC in bottle 6 was 2.0 mg/l. (*2 Maximum theoretical value = 59.8 mg/1)
Degradability based on the DOC measurements was not calculated
because
the test substance was not dissolved in the test solution. (Table 1 (P-19) and Table 2 (P-20)]
12.6 DEGRADABILITY BASED ON THE RESIDUAL TEST SUBSTANCE CONCENTRATION The test substance*3 was detected at concentrations of 101.3, 105.2, 91.5
and 106.7 mg/l in bottles 3, 4, 5 and 6, respectively. (*2 Initial concentration = 100 mg/1)
The degree of degradability based on the residual test substance concentration were 5 and 1 % for the bottle 3 and 4, respectively.
[Table 1 (P-19), Table 4 (P-22), and Figure 7 (P-32-33)] During condensation of the filtrate of the bottle 5 under vacuum, a part of the filtrate spouted from an evaporating flask. The spouted filtrate was recovered and then condensed with the remaining filtrate. However, the residual test substance concentration in the bottle 5 was obviously lower than those in the bottle 3 and 4 and it would impair reliability of biodegradability of the test substance in the bottle 5. Therefore, the degree of degradability based on the residual test substance concentration was not calculated for the bottle 5.
13
CONCLUSIONS
From the degradability results based on the BOD and the residual test
substance concentration, it can be concluded that the test substance is not
readily biodegradable and is not transformed* under the conditions prescribed
by the Modified MITI test.
ible 1 Summaryof the test results
Degradabilitieswere calculated by the equations shown in Sec. 11.7.
De adability based on BOD
ittle Test
THOD
Na
substance
mg
1.
aniline
90.2
2
---
3.
D-1
27.7
4
D-1
27.7
5,
D-1
27.7
6
D-1
27.7
Where % degradability was
da 7
da 14
BOD
% Degra-
BOD
% Degra-
mg
dability mg
dability
56.2 3.1 3.0 2.4 3.5
59 ---
0 -3
1
63.7 5.0 5.0. 4.11 5.71
T5 ---
0 -31
111
0.0
O-Of
calculated to be negative, this value is
day 21
BOD
% Degra-
mg
dability
66.3
67
5.5 - - -
6.1
2
4.81
-3
6.71
4,
shown in brackets.
da BOD mg
67.4 5.8 6.8 5.5 7.4 0.0
28 % Degradability
68 ---
4 -1 6 ---
I Oegradability base )tileOrganic carbon No. added, mg/l
3
28.8
4
28.8_
5
28.8
6
28.8
on DOC TC mg/l 1.7 2.5
2.11 3.31 2.0
ic mg/l
0.0 0.1 0.1 0.0 0.0
DOC mg/l
1.7 -2.4
2.0 3.3 2.0
d) Degradabilitybased on residuhl t
Bottle
Concentration Degra
No.
mg/i
2
<3
3
101.3
4
105.2
g --
91.5
6
106.7
not calculated because the test substance mi
accidentallyduririgcondensationof the solu
Table 2 Resultsof DOC measurements
Standgrdsolution
TC 50.0 mc/L
vlal :81 -------------
A arer -------------
1 346737 2 34 3 346490' -------------
MN
34690
TC 20.0 mg/L
vial :a2 -------------
# arer -------------
1 14107 2 139941
3 140 9 -------------
MN
14045
IC .10.0 mg/L
vial :63 -------------
# arer -------------
1
7185
-2
7107
3
7187
-------------
MN
7159
IC 0.0 mg/L vial :&4 -------------
# aror -------------
1
0
2
0
3
0
------------
MN
0
Bottle2,
TC ----------------------------
# arer
mg/L
RKK
----------------------------
1
1178
1.71
2 1157
1.68
3 1135
1.65
--- ------------------------
MN
1156
1.68
Bottle3
TC ------------------------------#---a-r-e-r-----m-e/-L-------RM-K----
1 1743
2.53
2 1.743
2.53
3 1713
2.49
---------------------------
b(N
1733
2.52
Bottle4.
TC ----------------------------
# arer
mc/L
RMK
----------------------------
1
1470
2.14
2 1418
2.06
3 1374
2.00
----------------------------
MN
1420
2.06
lc ----------------------------
# aror
mg/L
RMK
----------------------------
1
0
0.00
2
0
0.00
3
0
0.00
----------------------------
btN
0
0.00
lc ------------------------------0---a-r-o-r-.----m-s/-L-------RM-K----
1
0
0.00
2
0
0.00
3
166
0.23
---------------------------
MN
56
0.09
ic ----------------------------
# aror
me/L
RMX
----------------------------
1
157
0.22
2
0
0.00
3
0
0.00
----------------------------
MN
52
0.07
.Boftl5e
TC ----------------------------
# arer
mg/L
RMK
----------------------------
1 2340
3.40
2 2302
3.35
3 2215
3.22
----------------------------
MN
2285
3.32
Bottle6
TC ----------------------------
# arer
mg/L
RMK
----------------------------
1 1442
2.10
2 1326
1.93
3 1354
1.97
----------------------------
MN
1374
2.00
ic ----------------------------
# arer
mg/L
RMK
----------------------------
1
0
0.00
2
0
0.00
3
0
0.00
----------------------------
MN
0
0.00
ic ----------------------------
# arer
mg/L
RMK
----------------------------
1
0
0.00
2
0
0.00
3
0
0.00
----------------------------
MN
0
0.00
----------------------------------------------------
SPL
TC.me/L
RMK
IC.sx/L RMK
TOC.ne/L
Bottle2
2 1.68
0.00
1.68
Bottle3
3
2.62
4
2.06
0.08 0.07
2.44 1.99
<------ <
Bottle4
5
3.32
6
2.00
0.00 0.00
3.32 2.00
(
< ---- @@-@Boftle 5
----------------------------------------------------
Bottle6
Table 3 Calculationof recoveryand detectionlimit
Recovery test 1 Recovery test 2 ,Blank
Peak area AV-sec (A) 4805210 4815940 <132800
Concentration in
solution, mg/l
for GC in bottle
analysis
(B)
(C)
578.8
96.5
580.1,
96.7
<16.01
<2. 7
Recovered
Added
mass
mass
mg
mg
(D)
(E)
29.0
30.0
29.0 ----
30.0 0.0
Average recovery:
Recovery Detection
limit
% (F)
mg/l (G)
97 - - - -
97 - - - -
----
3
97 %
Standard solution
Concentration
Peak area
Volume of solution for GC analysis
Volume of test solutionfor GC analysis
Equation :B=HXA-* I C=BXJ-* K D=CXK
(H) (1) (1): (K):
F=D
500 mg/l
4150910 gv-sec 0.050 1 0.300 1
EX100
G=C raise decimal fractions to unit.
Calculationof detection limit
Retention time, min -5.6
6.2 65
500 mg/l std.
Peak area Rate of pea@
tLV-sec
area, %
298720
7.2
3127000
75.3
725190
17.5
4150910
100.
Blank Peak area
a V- see ---
<100000, --
<132800
The minimum detectable peak area of the'test substance was calculatedby the following equation based on the minimum detectable peak area of 100000 uV-see for the largest peak (retentiontime 6.2 min.) in GC chromatogram.
Minimum detectable peak area (,aV -sec)
100000 (.uV-sec)
4150910 X - - - - - - - - - (jLV-sec) 3127000 (g V-sec)
<
132745 132800
(,uV-see) (,uV-sec)
Table 4 Residualtest substanceconcentrationin test bottles
Bottle
No.
2 3 4 5 6
Concentration in
Peak area liv-see
solution, mg/l
for GC
in bottle
analysis
(A)__ _ <132800 5149880 -5349150
654900 54281201
(B) <15. 2 589.3 612.1 532.7 621.2
(C) <3 101.3 105.2 91.5 106.7
Standard solution
Concentration
(D)
Peak area
(E)
Volume of solution for GC analysis Volume of test solution for GC analysis Average recovery
(F): (G): (H):
Equation :B=AXD -E C--AXD EXF G--!--HX100
500 mg/l 4369340 4V-see
0.050 1 0.300 1
97 %
Figure 1 Infraredabsorptionspectraof the testsubstance
Measure by the-P.Pns-or
0.71
.-.963 $5.239
37,-X.00 79.M
7-@
3A4.-
nu. 0Do0
EM7. 00
0 "-g.co.0000
7A 512
SLZDD
U. 997
74A 2
.-p 4 --.
17=. 0: -
1139.
At.37:
4Z.7'-' 34.203 27.021
IAIM.:: !A&g.S5a17
sA4s:di r.n7.-0
!-.".00
in
1207.:: s4.oo
i 260 .7.7
9:495
U.113
n.u7-s7
EM.00 54. Do
24.710 31-M
21C.00 ts
sic.:::
757 a* 7-C7.0:
57.350
=.so =.Me
AE.W? A-- 9!3
739-.D M.A'B5
'S.ZV?ss ll.AF7
709.00 W-.Do 5= 254:.:1,:0
g3-n-..0
32.DCZ =.540
-IV amt: ff7 0 3X.200
3.=5
Acoo
me
2300 2000 Igoe 1200 goo
DotrZtorl
400
Measure@in M.S.I.
X@ 4n(In.0.4no.nii cm-t; o.ni, fit).5%3r
2:
02/ni/13 .11)"ClStrulig
00.31
4 scisnu: saild i*atiu-. I'@llut 4.Go L:
8363U 0-i Lut No.4 I
threstiuld
6.0n%*. tianct
cis -I
%
cm-1
,988.3
24.t7
1619.13 2U.11
itirl;1!. 7.-I-l
808.1
U.87
2364.1
i4sH.8
It)ty.A
ld4.1
50.05
18.77
i.sn
22.2ti
t729.3
13U2.0
C)IIA..4
'14Y.4
4.34
5.27
7. tf)
id.J4
A. Okuda
co-I 1639.4
1202.0
nifl.1
853.7
19.44
2.186 36.1
11.30
16 paeka found
T --.r--i H STt
C-H stretch
tc'. T C-C so
sttfC,e.hli@cch
5.0
0.07
$Goo
500
.000
3500
2000
2340
mto C--O stretch isoo
S--OstretchC-F stretch
C-0 stretch
Figure 2 Infraredabsorptionspectrum of the testsubstancestored in a refrigerator
X: 4000.0. 400.00 Cm-l-. 0. 14. 77.66 xr
4 scans: made ratio: resal 4.00 cm-1;
13363(2)0 0-1 Lot No.4
thr1edliald 5.OGX: bamd
cm7-1
co-I
21961 .2 51.57 9.8 54.81
2353.0 1468.8
1062.3 20.58
lotO.0
784.2 46.'59
747.2
x 68.77 40.04 17.95 40.04
cm-1 1731 .1 1392.9 985.1 735.8
16 peaks round
aped
92104116 strong
A.
3.85 5.36 16.75 43.83
cm-1 1636.8 1205,5 808.1 653.6
16.39
OkUde
x 42.99
1.32 25.32 25.40
C-H stretch
t stretch
4000
35@o
3000
T 20(1 C--O stretch ISO
1000
co@
S--o stretch CF stretch
C,o stretch
Figure3 CalibrationCurve ofthe Test Substance
CalibrationCurve.
Concentration X(mg/1) 0 250 500 1000
Peak Area Y(YV-sec) 0 1.92785E+06 3.89009E+06 8.12341E+06
Y= -7.552xlo4 + 8.139xlo3 X r= 0.999707
X106 10
8 L(U)
a) L-
41 CL-
2
oiz 0
500
1000
eoncent-rati-en(mg/1)
ir
IF
0 c-6.499
to
(D
CD 0
AREA%
R7
5 60
5.88 6.29 6.32
6.48
11.09
AREA TYPE
pgt7ea n pp
156388 0 py
2885188 0 VY
188128 0 VY
713218 D Vg
2819600
PY
AR/HT
A-ago
0.898 9,090 0.964
R@871 1.335
CD
AREA%
4 13(;
2.217
48,999 2.660 le@tio 39.9/19
3 Ati IF ct
IF
-----I
6.24
co
1.63
0
T
11.09
AREA%
Ri
AREA TYPE
AR/IIF
4. tl3
5.06 5.32 .1-61 5.89
100228 0 VV
127970 0 va,
117448 D PP
Out.@ PF 318488 0 PV
9.090
0.886
0.676
0,103 0.997
6-50 fi.99
1412506-0 YO 8@876 534090 0 BY 0.26,
CD =r < ID
0
CD
AREA% 1.154
1.988 1.252 6-IBB 3.396
15-1 5 5.695
51ARI IF l@IFF
Nd RUH PEAKS STORED
s ARI IF
et
I
IF
.46
to -4
11...884969-
AREA%
R7
6,176.46 lt.09
AREA TYPE 145 pa 0 BP 143 08 D fly
2998400 By
AR/HT
0 896
0-078 --29.02JL
1.678 60.865
IF rt 0 ft
6.46
0
<
7613
(D
(D
co
AREA7
Ic@-
RI
AREA TYPE AR/HT
AREA%
5. t35
144480 D VV 9.185
1.606
5@60
362918 0 VP 0 - 100
4,035
5.08
194960 0 PV 0.892
2.167
6.21
3565700 D VY Oz 895
39,641-
6.33
232290 0 VV 0.864
2.598
9-48
826680 0 VB 0.072
9.ZI5.
-f.63
618298 VY 9.756
6.874
lt.88
29993tio VY 1.182
33.344
SIARI IF
-71
0rt,
IF
a
I0[D
C()D
i@s6o cc
ko
1 IL
0
AREA%
RT
AREA TYPE AR/HT
AREA%
5.96
104890 0 Po 0.084
1.269
5.60
360210 p pp 8 R99
4-3'iq
5.88
195250 0 PV 8.092
3572709 n vy 0 R99
2.363 43-315
6.33 -E.48
232248 0 VY 9.064
8768 n n yn 0-023
2.818 IR 6AR
11.88
2915908 BY 1.258
35.284
SIARI IF IF 11*.09 6.41
AREA% RI 5.06 5 68
11.09
AREA TYPE 105999 .0 vy
296720 D yp 1617 0 py
AR/HT
0.896 e.e99 9.091
AREA% 1.664 4.609 -F53-8
3juni-ELit L-JLSM---12-u--L
t9tiz/vo . ry
871 1.091
38.648
SIARI IF
F
110 RUH PEAKS
STOREO
Figure6 BOD chart
.0
E -4 5--
U) 0
-3 0-
CYL--IM
7-
MeaSUTement of BOD
Test substance: D-2
Study nwaber
IB363(2)G
Period
3/18/1992
to 4/15/1992
Temperat ure
2.5t I 'C
Apparatus
Ohkura Electric Co.
Model OH-2001(l.D.No.:B)
Chart speed : 2mm/hour
bottl Subs tance Cone. isludmgge/clonc.
No.
mg/l
1
Aniline 100
30
2 ------
----
30
3
D-1
100
30
4
D-1
loo
30
5
D- I
loo
30
256
D-1
Operator
100 Ayumi Okuda
-------. ........
---.. ....
0
-- 0 1 da)
--7
(hour)
Li :Bottl
..
...........
Bottle
Figure6 (Continued)
Bottle1
!Percentagedegradabilityof anilineat day 7
BOD in bottle1
BOD in bottle2
56.2 mg - 3.1 mit
X 100
59%
90.2 mx t
THOD ofaniii
- ---------------------------------
Bottle5 Bottle2 Bottle3 Bottle4
Bo@le 6
'igur6e (Continued)
14 1
I .(336)
. I;
.
I
Bottle1
17
.........-...
Bottle5 Bottle3 ':Bottl2e Bottle4
Bo@le 6
.'igu6re(Continued) .........
28
Figure7 GC chromatograms of the testsubstance - Measurement of residualtestsubstance concentration The concentrationsofthe residualtestsubstance in the bottlesare shown inTable 4.
500 mqA in-ethylacetate
uVajc-eoM:;- r-n0-Iinn IX . IV:o@
0Ch0C0%U NON%wo-CmU
m CD 0 co co c co c@ W, c@
ui CL Z-.2-. m
0
c:i
a CD m C'D M crS,)
LLir, (U = = &p w -w CD
4= 0 OD:; = CU
cu N cu
m
r.)-
cu w
C%i cc C%i @ 04
'i@c@IU li
L61
W@%n
cr
Cc
.total peak.areat436934ORv*sec
Bottle2
xx 40 I.U) U) LU
ix
Rottl63
Umi r@ am-WMM
C@ ri
L,5CD eu m m wt
c@ 'cr c@ to' c@ c@ c@ CD'
> CL CL >. >I--ca
CgTwg:ww W%
a tn
12.@ll> cc
ON
V)
-,a-
co
m m ad 4
vlo CP w
. 0
63 01 cu
C@ in
total peak area:51498801LV*Sec
ccC%i%a --r 0 N
C%J
%a r.)Ch X)
I.- co
ru
c@ c@ c@
> CL.> -@- c-
c= = C2 0
= co CwD CD w cc r,
w P,*Pcm
ON
CD co Ch
m
cr. r,
W)
cCoj
CD
C%i C0D,
V'j Lfo
C@ %D C-ul
t6talpeak areat5349150@tv'sec
STARI IF
IF
3
6.49
6.22
AREA% RI 4.73 5.03 5.62 5.98 6.22 6.34 6.49
AREA TYPE 322608 0 PY 164988 0 VY 33 193;19: s ;yp
347271, E26410 0a VyVy 846999 D VO
AR/HT
8.985
0.183 '181
:.99Z .093
10.864 0.072
AREA% 5.818 .2.971 6.g@19 i.ili! 6Z.546 4.U77 15.251
5 ARI IF et
F@IIF
Im
co
AREA% RT
S.62 5.90 6.22 c qA
AREA 361488 zuuuiu 4159208 907448
TYPE 0 PP u ry
VV u Yo
AR/HT
AREA%
0-100
5.850
F.093-3.258
0.163
67.386
8.872
14.bU@
18
8.!)FB-