Document g25kzp5EwzZw3p15jE4m64VLL
BIOACCUMULATION
TEST SUBSTANCE
Identity:[2-(N-Ethylperfluorooctanesulfonamiedtoh)ylacrylate;may also be referredtoas Bl 228, D-1, ETFOSEA, or FX-13.(2-Propenoicacid,2[ethyl[heptadecafluorooctyl)sulfonyl]aminoe]settehry,ClAS # 423-825)
Remarks:Materiiaslan ambersolidR.eportstatetshatsamplepuritiys99% or more based on informatiosnuppliedby Sponsor. The pudtyfidentitoyf the testsubstance cannot be substantiated.As presentedinthe report,the structuraflormulaindicatesthat"purityc"annot be assigned as "99% ormore. Lot number 101.
METHOD
Method/guideline followed: MITI describedin"Chemical Substances Control Law" (Japanese Law No. 117,1974). OECD 305C. GLP (YIN): Yes Year: 1995 Exposure period: 8-weeks Exposure concentrations:0.01and 0.1mg/L Test species: Cyprinuscatpio
RESULTS
BloconcentrationFactors:
Week 2
0.01 mg/L exposure:
0.4
duplicate
0.4
0.1 mg/L exposure
3
duplicate
3
Week 4 0.6 0.7
4 3
Week 6 0.6 0.4
2 2
Week 8 0.5 0.5
2 2
CONCLUSIONS
Neithermortalitnyor abnormalitiesinappearance orbehaviorwere observed in the treatedand controlfishduringthe 8-week testperiod.
The testsubstance isnot bioaccumulativeincarp under the prescribedtest conditions.
Submifter:3M Company,EnvironmentLaalboratorPy.,O.Box 33331,St.Paul, Minnesota, 55133
DATA QUALITY
ReliabilityK:limischranking3. The pudtyridentiotfythetestsubstance cannot be substantiated.The extraction/analytimceatlhodology has notbeen validatedthereforeitisnot possibleto know ifthe testcompound didnot bioconcentrateinthefishor simply was not extractablbey the methodology ofthe experiment.The reportstatesthat the chromatographicpeaks elutingfrom "l to3 minutes"are consideredtobe metabolites.While thisstatementdoes notchange the conclusionsofthe report, thisassumption iscannot be justified.
OTHER
Last changed: 5/22/00
M.S.[.
REPORT
No.2BI21G
BioaccumulatSitoundyofSamplbe-I
with Carp ( Cyprinus carpio
Submitted to SUMITOMO 3M LTD.
Prepared by MitsubishiChemicalSafety InstituteLtd.
Nov. 30, 1995
a
AROMA
ab
TRANSLATION OF TIE TEST REPORT
Study No. 2Bl2lG
Study Title BioaccumulationStudy of Sample D-1 with Carp
carpio)
Study Period From May 18, 1992 to Sept. 24, 1992
Sponsor
SUMITOMO 3M LTD.
Cyprinus
This test was conducted in Yokohama Laboratory, Mitsubishi Chemical SafetyInstituteLtd.(M.S.I.),1000 Kamoshida-cho,Aoba-ku, Yokohama 227, JAPAN.
The originaltest reportwas writtenin Japaneseand this reportwas translated into English. The persons,the undersigned,herebydeclare that this reportreflectsfaithfullythe originalreportas much as possibleto our knowledge.
Translatorfrom originalJapaneseversion
-D.
Date
Chief ResearchScientistof EnvironmentalSciences Division
....................... ....
............................
Shoko TANIMM, B.Sc.
Date
Scientistof EnvironmentalSciencesDivision
Approved by
Jun@oRi,Yi;A, Head of Yokohama Laboratory
9 Date
COMPLIANCE WITH GOOD LABORATORY PRACRICE STANDARD
Study No. Study Title
Sponsor
2Bl2lG BioaccumulationStudy of Sample D-1 with Carp
carpio) SUMITOMO 3M LTD.
Cyprinus
To the best of our knowledge and belief, the study described in this
report was conducted in compliancewith the Good LaboratoryPractice (GLP)
regulationsrecognized by Basic IndustriesBureau, Japanese Ministry of
InternationalTrade and Industry(MITI). Yokohama Laboratory, MitsubishiChemicalSafety InstituteLtd. (M.S.I.)
The company's name was changed on October 1, 1994. Former name Mitsubishi-kaseIinstituteof Toxicologicaland EnvironmentalSciences
Test Facility Managemen Sakae KOIKE, B.Sc. Head of Yokohama Laboratory
Sealed Date Sept. 24, 1992
Study Management Tadayoshi SHIGEOKA, Ph.D.
Sealed Date Sept. 24, 1992
Chief Research Scientistof EnvironmentalSciences Division
Study Director
Ayao NISHIKAWA
Sealed Date
Scientist of EnvironmentalSciencesDivision
Sept. 24, 1992
We, the undersigned,declare that the work was performedaccording to the procedures herein described,and this reportprovidesa correctand faithful record of the resultsobtained.
ExperimentalScientist
Yasuo SATO
Sealed Date
Scientist of Env.irorunentSacliences Division
Sept. 24, 1992
.Naomi SAEKI, B.Se.
Sealed Date
Scientist of EnvirormentalSciencesDivision
Sept. 24, 1992
QUALITY ASSURANCE STATEMENT Yokohama Laboratory, MitsubishiChemicalSafety InstituteLtd..
Study Title BioaccumulationStudy of Sample D-1 with Carp (Cyprinus'carpio) Study No._ 2Bl2lG
.We hereby certify that the above study was performed in compliancewith the Good Laboratory Practice (GLP) regulationsrecognizedby the Basic Industries Bureau, Japanese Ministry of InternationalTrade and Industry (MITI).
Dates of inspectionsand findingsreportedto the Study Directorand the Facility Managementare as follows:
In-progressstudy Final report
Dates of Inspection
May 26, 1992 June 23, 1992 July 21, 1992
Sept. 24, 1992
Dates of Report;lng
May 26, 1992 June 23, 1992 July 22, 1992
Sept. 24, 1992
Quality AssuranceStaffs Junko KATOH, Ph.D. Yumiko MIURA, B-Sc. Fumio YAMAUCIII,Ph.D.
Sealed Date : Sept. 24, 1992 Sealed Date : Sept. 24, 1992 Sealed Date : Sept. 24,'1992.
T a b Ie o f C o n t e n t s page
Summary ...........................................................8.............. 1 Introduction....................................................9..............
1.1 Studytitle
9
1.2 Objective
9
1.3 Testing Method
9
1.4 GLP compliance
9
1.5 Study Period
9
1.6 Storage and retentionof recordsand sample
9.
.2 Test Substance........................................................1.0..............
2.1 Chemical name, Structural formula,Molecularformula and weight 10
2.2 Physico-chemicalproperties
10
2.3 Source
11
2.4 Confirmationof test substance
11
2.5 Stabilityof test subsanceunder storageconditions
11
3 Materials......................................................1.2.............
3.1 Apparatusfor bioaccumulationtest
12
3.2 Apparatusfor analysis
12
3.3 Reagents
12
4 Analyticalmethods -:............................................1.3.............
4.1 Gas chromatographic-(GC)6onditions
13
4.2 Calibrationcurve
13
4.3 Analyticalmethod of test substancein testwater
14
4.4 Analyticalmethod of test substancein testfish
15
4.5 Blank test
17
4.6 Recovery test
17
4.7 Detectionlimit
18
5 Acutetoxicityto fish ..........................................19..............
5.1 Preparationof test solutions
19
5.2 Test conditions
19
5.3 Results
20
6 Stabilityof test substanceunder the testcondition.s...............2.1..........
6.1 Stabilityin water
21
6.2 Stabilityin feedsolution
21
7 Bioaccumulationtest method.......................................23..............
7.1 Test fish 7.2 Methodsand conditions
23 23- 7-
8 Results and Discussion .........................................2.6..............
8.1 Circumstancesthat may have affectedthe reliability
of the test results
26
8.2 Biologicalobservationsof test fish
26
8.3 Concentrationsof test substance in test water
26
8.4 C6ncentrationsof test substancein test fish and bioconcentration
factors(BCFs)
26
Table 1 -7 and Figures1 -18 Appendix-1.Infaredspectrumof test substance
(Data:providebdy the sponsor) Appendix-2 GC chromatograms(Analysisof water)
28-55 .1 page
17 pages
Summary
Sponsor
: SUMITOMO 3M LTD.
StudyTitle : Bioaccumulatiosntudyof-SampleD-1 with carp Cyprinus carpio
Study No.
: 2Bl2lG
Test Substance : Sample D-1
Test Period : From May 18i 1992 to Sept. 24,'1992
Testing Method : Testingmethodof Ministryof InternationaTlrade and Industry (MITI) described in "ChemicalSubstances Control Law" (JapaneseLaw No.117, 1974). (OECD testingguideline305C) Supplyof testwater : Continuousflow-througdhiluter system flowrate : 1600L/ day Nominalconcentrationof SampleD-1 in testwater High exposurelevel ; 0.1 g g/mL Low exposure level ; 0.01 g g/@nL Exposureperiod : 8 weeks Analyticalmethod : GC analysisafterpre-treatment
Results
Bioconcentratibnfactors(BCFS)obtainedin thisstudy were as follows:
Bioconcentratiofnactors (BCFs
level-"@@exl@sure period(weeks) 2
4
6
8
High exposurelevel,samplefish No-1 0.4 0.6 0.6 0.5 No.2 0.4 0.7 0.4 0.5
Low exposurelevel,samplefish No.1 3
4
2
2
No'.2 3
3
2
2
Conclusion
On the basisof the above results,it can be concluded that the test substanceis not.bibaccumulativuender the
n t rod u ct ion
1.1 StudyTitle BioaccumulationStudy of SampleD-1 with Carp ( Cyprinus carpio
1.2 Objective The objectiveof this studywas to evaluatebioaccumulatiopnotential
of D-1 for applicationof a new chemicalsubstancein Japan. 1.3 Testing Method
Testingmethodof Ministryof InternationaTlradeand Industry(MITI) described in uchemicalSubstancesControl Law" -(JapaneseLaw No.117, 1974). (OECDtestingguideline305C) 1.4 GLP Compliance
The study was conducted in accordance with the Good Laboratory Practice (GLP) regulationsrecognizedby Basic IndustriesBureau, JapaneseMinistryof InternationaTlradeand Industry(MITI). 1.5 Study Period
From May 18, 1992 to Sept. 24, 1992 exposureperiod.:From May 26, 1992 to July 21, 1992)
1.6 Storage and retentionof recordsand sample Followingrecords,raw data, and samplewill be retained in the
archivesof MITES for 10 yearsaftersubmissionof the final report. After this period,the sponsorwill be contactedfor approvalof the disposalof any data. Data will be retainedin the archivesfor a furtherspecifiedperiodat the sponsor'srequest(additionaflee). 1) The protocoland its ammendmentrecord. 2) The final'report. 3) Raw data. 4) Qualityassurancereports. 5) Testsubstance( ca. 2g Ai nthi-rdnriimentrsea'uiredbv'MITI OLP'i6k@l-itons.
2. T e s t s u b s t a n c e
2.1 Chemicalname,Structuralformula,Molecularformulaand weight. Chemical name 2-[N-Ethyl-N-perfluoroalky1l-(8C)=sulfonylaminolethylacrylate [Abbreviatendame:SamleD-1, identificatioNno.2Bl2lG]
Structuralformula Cz H5
C. Fz..iSOt
0 %'OHzO-C-CH=CH2
n = 1-8, n 8 : ca. 78%, n 1-7 ca. 21%
Molecularformula Ci5HizO4 NSF17
Molecularweight
6 2 5 (n= 8)
Data from the sponsor)
n8
2.2 Physico-chemicaplroperties
Appearance
amber like wax
Meltingpoint
27 -420C
Boilingpoint
ca.150 OC ( ImmHg)
Solubility
water
< 0. 1 % 25 *C)
methanol
> 5 % 25 OC)
acetonitrile > 5 % 25 OC
acetone
> 5 0%
n-hexane
> 5 N 25 *C)
THF
> 5 % 25 -C)
DMSO
insoluble
Infraredspectrum* shown in Appendix-I
Data fr6m -theg@6h's'-o-rl'-
7-
2.3 Source
(1) Supplier
: SUMITOMO 3M LTD.
(2)Suppliedquantity* : 50 g
(3) Date of receipt
: April 1, 1992.
(4)Lot.No. *
: 101
(5)Purity
> 99 %
Impurity*
Phenothiazine 60 20ppm
Hydroquinone monomethylether 170
provided by the sponsor
35ppm
2.4 Confirmationof testsubstance Before starting the study, infrared (IR) spectrum'ofSample D-1 was
measured. The spectrum (Fig.l(p.28))was compared with the IR data provided by the sponsor (Appendix-1)and was confirmed the identity. And it also correspondedto the structuralformulashown by the sponsor.
2.5 Stabilityof test substanceunder storageconditions At the termination of the study, IR spectrum of Sample D-1 was
measured and the spectrum (Fig.2(p.28))was compared with the IR spectrumobtainedat the start of this study (Fig.1).Test substancewas confirmed to be stable under storage conditions stored in the refrigeratorduring the test period of ca.2.5months.
3. M a t e r i a I s
3.1 Apparatusfor bioaccumulatiotnest The flow-throughtestsystem is shown in Fig-3 (p.29 f For testaqruariumI Dilutionwater supplypump :Yamazen (FMI) Co., QD-2CSC Feed solutionsupply pump :Yamazen (FMI) Co., QG6-FIfOOSTY [ For controlaqruariumI Dilutionwatersupply pump :Yamazen (FMI) Co., RP-D-2CKC Feed solution supply PUMP- :Tokyo Rika Kikai Co., GMW-8
3.2 Apparatusfor analysis Infraredspectrophotometer Homogenizer Shaker Rotary evaporator Aspirator Electronicbalance Gas chromatograph
: Parkin Elmer Co., Model 1640 : NihonSeikiKaishaLtd.,M-8 : Taiyo Kagaku Kogyo Co., SR-II : BuchiLtd., RE-111 : Yamato Kagaku Co., BP-51 : MettlerLtd.,PL1200 : Shimadzu Ltd., GC-14A
3.3 Reagents
Acetonitrile
Kokusan Chemical Co., GuaranteedReagent
Acetone
Junsei Chemical Co., GuaranteedReagent
Chloroform
Junsei Chemical Co., Guaranteed Reagent
Ethyl Acetate
: Kokusan Chemical Co., GuaranteedReagent
Tetrahydrofuran(THF) : KishidaChemicalCo., GuaranteedReagent
n-Hexane
: Kishida Chemical Co., GuaranteedReagent
Methanol
: Junsei ChemicalCo., GuaranteedReagent
Sodium chloride
: Kishida ChemicalCo.-,GuaranteedReagent
Sodium sulfate,anhydrous
:
i(okusanOh.em"ic-aC-of.,
Guaranteed
-. Reagent
4. A n a I y t i c a I m e t h o d s
4.1 Gas chromatographic(GC)conditions
Column Shimadzu Ltd.,Wide Bore column, CBP20-W25-100
Temperature
Column
1300C
Injector 2000C
Detector
2400C
Flow rate of N2
20mL/min
Range
10'
Attenuation
7
Chart speed
5mm/min
Injection volume
3gL
0.53mmx 25m
4.2 Calibrationcurve Standard solution of 1000 ag/mL was prepared by dissolving100 mg of
D-1 in 100 mL of n-hexaneand this standardstock solutionwas diluted with n-hexane to prepare 0.25, 0.50, 1.00gg/mL n-hexanesolutions of D-1. These solutions were injected into GC describedabove. The GC peak areas( g V -sec)-were plottedagainst the concentrationsof D-1. The GC chromatogranfsand the calibration curve thus obtained are shown in Fig.4(p.30),and Fig.5(p.31),respectively. Quantityof D-1 was calculated from the total area of peaks at 3.9 and 4.5 min. on the chromatogram. The calibrationcurve is sttaight line throughthe origin and its correlationcoefficientis 1.000.
D-1 was determined with the ratio of area on GC chromatogramsagainst the standardsolution (0-5 gg/mL) per measurement.
4.3 Analyticalmethodof testsubstancein testwater Analysis was performed accordingto the followingprocedure.
Test Water 1000 mL (100 mL) * I
+---Sodiucmhloride 25 g,(5g *--Chloroform 80 mL 40 mL) -Extractwith shaker (5min.)
Aqueous 1
+- Chloroform 80 mL( 40mL I Extractwith shaker (5min.)*
Chloroform laye7r
Aqueous layer
[-Chloroformlayer-]
Discard
Sodium sulfateanhyd. 40 g Filter ( llG3glassfilter
Evaporate to dryness
N-Hexane Dilute to 20mL
GC
............................................................................. High exposure level
4.4 Analyticalmethodof test substancein test fish Analysiswas performedaccording to the followingprocedure.
Repeat once again
Test Fish (ca. 25g
Wh
Cut into ieces *.-Acetonitrile8-0 mL
Homogenize, 5 min. 10,000 rpm) Filter suction Filterpaper S-60)
Residue
Acetonitrilelayer
L Discard
+-@ionized water 5OOmL
*-Sodium chloride 25 g
n -Hexane
IOOML
Extractwith shaker,5 min
Fcetonitrile-aqueous l@ay@er]
n-Hexane
Discard
*--Acetonitrile 80 mL
Extract with shaker,5 min.
n-Hexane later] Discard
Acetonitrilelayer
-Evaporate to dryness +-n-Hexane 5mL
Dissolve
Continued on next page]
n-Hexane solution 5
Column chro tography Elute
SEP-PAKTM cartridge(Florisil) Waters Co.
Eluent
Column .+-n-Hexane/EthylAcetate=8/2, lOmL
Elute
Eluent
L'@-Hei +-n-Hexane Dilute to 20mL I GC
Column
Discard
. ..
4.5 Blank test (1) Water The blank tests were performed twice according to the procedureshown in 4.3, using 1000 mL of the test water in the absenceof D-I(Control aquarium). There was no peak which disturbed the analysisof D-1 in GC chromatogramsfrom water(Fig.6(p.32)). (2)Fish The-blank tests were performed according to the procedure shown in 4.4, using the each two untreatedtest fish with D-1 (Controlaquarium) ,at the bigiming and the terminationof the exposureperiod. There was no peak which disturbed the analysis of D-1 in GC chromatogramsfrom fish(Fig.7(p.3334,)).
4.6 Recovery test (1)Water Two mL of acetone solution of D-1 (5gg/mL) was added to 1000 mL of the test water in the absenceof D-1. D-1 dispersesolutionof 0.01 gg/mL (the same as Low exposurelevel concentrationin the test water) in the test water was thus prepared and the recovery-tests were performed twice according to the procedure shown In 4.3. As shown in Table l(p.35)andFig.8(p.36),the mean value of the recoveryratioswas 94.6%. Analyticalvalues of the test water were correctedby this value. (2)Fish Two mL of acetone solution of D-1 (5gg/mL) were added to the untreated test fish with D-1 and the recovery test was peformedtwice according to the procedureshown in 4.4. In case that a fish body weight is 25g, the concentration of D-1 in fish would be 0.3g g/g and correspond to 30 times of that in the test water. As shown in Table 2 (p.35) and.Fig.9 (p.37),the mean value of the-recoveryratios was 99.2%. Analyticalvaluesof thd-t&:tf1-S''h,Wdecdorrectedby.thisvalud.-7-
4.7 Detectionlimit There was no peak which disturbed the analysis of D-1 in GC
chromatograms from blank water and blank fish. Therefore, the area of
the minimum detectable peaks, which can be recognized visually, was set
to be 30,000 gV See.. This value correspondsto ca. 0.02 g g/mL(D1) on the calibrationcurve. Detection limit is calculatedaccording to following calculation formula and conditions. The results are as
folloi4s; Fish Water
: 0.02gg/g : High exposure level
Ww exposure level
0.004 g g/ML 0.0004,4g/ML
It would be possible to calculate bioconcentrationfactors (BCFS) of
following values in the case of fish weight 20g.
High exposure level ( 0.1gg/mL )
> 0.2 times
Low exposure level ( 0.01 g/mL )
> 2 times
...............................................................................
(Calculationformula]
Detection limit Concentration in x Volume of the /Amount of the final solution final solution sample
[Conditions] Fish
Volume of the final solution Amount of sample (fish weight)
20 mL 20 g
Water High exposure level ; Volumeof the final solution 20 mL
; Amount of sample
100 ML
Low exposure level ; Volume of the final solution 20 mL
Amount of sample
1000 ML
5 . A c u te tox ic ity to f ish
The staticacute toxicitytest of the test substanceto fish was conducted according to the Japanese IndustrialStandard Method "JIS K 0102-1986, IndustrialWaste Water Testing Method, 71. Acute toxicity study using fish". The result obtained in this test were used as a reference for determining the nominal concent-rations to be used in the bioaccumulationtest because the nominal concentrationsof high and low exposure levels should be lower than 1/100 and 1/1000 of 48hr-LC50, respectively-
5.1 Preparationof test solutions Four mL of 400mg/mL THF solution of HCO-20 was mixed with 4mL of
50mg/mLTHF solutionof D-1. After evaporationof THF in the resultant solution by blowing Na gas,it was mixed with HCO-20 solution(6.4gin lOOmL water) and dispersed.This dispersedsolutionwas diluted to 2L with declorinatedwater (Yokohamamunicipal tap water was treated with activatedcharcoal)toprepare 100 gg/mL solutionof D-1.
(controllevel Eight g of HCO-20 was dissolved with 100 mL of deionized water and diluted to 2L with dechlorinatedwater.
5.2 Test conditions
(1)Test fish Orangekillifish(Oryziaslatipes
Source
Miyazawa Fish Farm(1323, Suenaga, Takatsu-ku,
Kawasaki)
Date of receipt March 3, 1992.
Lot No.
92-H-0303
Body length
approx. 2 cm
Body weight
approx. 0.2 g
(2)Acclimation:
Aquarium No.
C-5
Water temperature
252 OC
Period
Over one week
Feed
Tetramin (D(TetraWerke, Germany)
Fish were fed once a day except holidays and
not fed 48 hours prior to the test.
Percentageof mortal fish during one week befor test ;lessthan 5%
(3)Test conditions
The number of fish 10 per one aquarium
Volume of water
2L
Temperature
24.4-25. 40C
Duration
48 hours
Dissolved oxygen
4.2-7.4 ppm
Aeration
Test water was aeratedduring the test period.
Renewal of test water non
5.3 Results The 48hr-LCso valueof test substancewas more than 100 g g /mL
as shown in Table-3*(p35).
6. S t a b i I i t y o f t e s t s u b s t a n c e t h e t e s t c o n d it io n s
unde r
6.1 Stabilityin water (1)Preparationof test solution
The feed solution of high exposure level (concentrationof'D-1: 4.Omg/mL)was preparedby the procedure in 7.2(l).Then, 2.5mL of this solution was diluted to 10OOmLwith delonizedwater..It(concentration of D-1: 10 gg/mL) was used for the stabilitytest of D-1 in water. (2) Method of measurement
The solution prepared by the procedure shown in 6.1 was left under room conditions, and 5mL of this solution was withdrawn and diluted with 100 mL of water on days 0, 1, and 2. Each of them was pre-treated by the procedure in See.4.3 and diluted to 100 mL with n-hexane.The resultant solution was analyzed by GC and variation of the test substance concentrationwith time was evaluated.
(3) Results Typical GC chromatogramsare shown in Fig.10(p.38)and the variation
of concentrationis shown in Fig-11(p.39). Based on the result that test substance remained 104% (9.5 g g/mL --,9..9gg/mL) of initial concentrations-after2 days, it is concludedthat D-1 is stable in water.
6.2 Stabilityin feed solution (1)Preparationof test solution
The feed solutionof low exposurelevel (concentrationof D-1: 400g g/ mL) was prepared-bythe procedurein 7.2(l). (2) Method of measurement
The test solutionpreparedb3(-(I)w,a8"leftunder the rdom.conditiont@
At days 0, 3, 7, and 14, 2.SmL of the test solutionwas withdrawn and was diluted to 50 mL with acetone. Then each of 2.5mL of these solution was withdrawn and was diluted to lOOmL with acetone, and analyzed by GC. Variation of the test substance concentrationwith time was evaluated.
(3) Results Typical GC chromatogramsare shown in Fig-12(p.40)*andthe variation
of concentrationis shown in Fig-13(p.41). Basedon the result that test substance remained 99% (394,ug/mL - 390#g/mL) of initial concentrationsafter 14 days, it is concludedthat D-1 is stable in feed stock solution.
7 . B io a c c um u Ia t io n t e s t m e t h o d
The study was performed in accordance with the testing method of Ministry of International Trade and Industry (MITI) described in "Chemical Substances Control Law" (Japanese Law No.117, 1974).
7.1 Test fish Carp( -Cyprinus carplo ) were purchased from Sank-yoSuisan Co. (1-1
Ichigayatamachl,Shinjuku-ku, Tokyo) on December 6, 1991. Prior to the initiationof the test, these fish were acclimatizedover a week under the.test condition (water temperature 25 2 *C, fed with commercial pellets once a day) and the mean body length of them is ca.11 cm. Mortal fish were less than % during the acclimation of a week prior to the introductionof fish into the test system.
7.2 Methods and conditions (1) Preparationof.feed solution High exposure level The stock solution was prepared by dissolving 2.0 g of D-1 and 80 g of HCO-20 in 5OOmL of THF. Cone. of D-1 4.0 mg/mL, Cone. of HCO-20: 160 mg /mL Low exposure level Fifty mL of the feed solution of high exposure level was diluted to 500 mL with THF (Conc. of D-1 0.40 mg /mL, conc. of HCO-20 : 16 mg/mL). -Control aquarium The 160g of HCO-20 was dissolved in IL of THF(stocksolution). The 370mL of this solution was diluted to 11 L with deionized water. (Conc. of HCO-20 : 5.4 mg /mL The feed solutions were renewed every 7 -10 days.
(2)Preparationof test solution The feed solution prepared by the procedure shown in 7.2(l) was
supplied to mixing tube by feed solution supply pump and dilution water (dechlorinatedtap water) was also supplied to the same tube. Then these were mixed and the concentrationo-f test substance reached nominal value. The test solution in mixing tube was supplied to an aquarium. Dilutionwater is the tap water of Yokohama city which was dechlorinated with activatedcharcoal.(Fig-3(p.29))
(3) Conditions
Flow.rate of feed solution 40m Uday for high and low exposure levels
and 1.2 Uday for control level
Flow rate of dilutionwater : 1600 L/day (The turnover rate for each
aquarium : 20 times/ day)
Test fish : Carp (Cyprinus carpio, Lot.No. 91-K-1206,aquarium No. B5)
Body weight
: 31-40 g
Body lengtb
: 11- 12 em
-Fat contentof fish : 3.7 % ( n=4, 3.3- 4.3%)
-Water temperature : 24.2-24.70C,
-Dissolved oxygen : 4.9 - 8.2 mg/L
-Feeding
Fish were fed with commercialpellets
(MinipetID(KyoriCno. Ltd.)) once a day exceptholidays.
-Nominal concentrationof D-1 in water
High exposure level ; O.lg g/mL (conc. of HCO-20, 40@/ML)
Low exposure level ; 0.01 g g/mL.(conc.of HCO-20, 0.4mg/mL)
Control
; 0 gg@ml (conc. of HCO-20, 4mg/mL)
-Populationdensity(at the biginningof the exposureperiod)
High exposure level ; 15 fish/ 80L testwater
Low exposure leve ; 15 fish/ 80L testwater
ig fich i AnT. toot tvotar
Exposureperiod : 8 weeks
(4) Introductionof fish, biologicalobservationand environmental control
The test system was operatedfor 4 days prior to the introductionof fish to confirmthe stabilityof the concentrationof D-1 in water. After that, fish were introducedinto the test system. Fish were observedfor mortality,behaviorand some abnormalitiestwicea week. Dissolved oxygen and temperature of test water were monitored twice a week. These results were recorded.
(5)Analysisof D-1 in testwater Aliquot of test water was withdrawn twice a week and was analyzed by
the procedure shown in 4.3.
(6)Analysisof D-1 in testfish Three fish were.collectedin 2, 4, 6 and 8 week after exposure. Two
of them were analyzed by the procedure shown in 4.4. Flow rate of dilution water was 800 L/day in first 3 days , but
because of concentrationdecreaseof D-1 due to uptake by fish, it was changed to 1600 Uday after the day for each aquaria.
8. R e s u I t s a n d D i s c u s s io n
8.1 Circumstancesthat may have affectedthe reliabilityof the test results. There was no significant matt-erthat may have affected the reliabilityof the test resultsduringthe test periodof 8 weeks.
8.2 Biologicalobservationsof test fish Neither mortality nor abnormality in appearanceor.behavior was
observed in treatedand control fish during the test periodof 8 weeks.
8.3 Concentrationsof test substancein test water The resultsare shown in Table 4, 5(p.42,43) and Fig-14,15(p.44).
(Appendix-2) The mean values during the exposureperiod were 0.092 g g/mL ( high exposurelevel ) and 0.0082 gg/mL ( low exposurelevel
The decreasesof D-1 concentrationin both levelswere observed at 3 days from the initiationof test. This phenomenawill be attributable to uptake of D-1 by fish. Therefore,flow ratesof the feed solution for both aquaria were increasedby a factor of two, and as a result, each concentration is.approached to each nominal value. The coefficientsof variationare 8.4 % ( high exposurelevel ) and 13.5% low exposurelevel ), respectively.
8.4 Concentrationsof test substance in test fish.andbioconcentration factors(BCFs)
The resultsare shown in Table6, 7 (p.45,46) and Fig.16-18 (p.4755). Concentrations'of D-1 in fish are 0.032-0-062 g g/g high exposure level ) and 0.013 -0.034 gg/g (lowexposure level The BCFs are 0.4-0.7 high exposure level) and 2-4 low exposure IPvp. I
Peaks observedat the retentiontime from 1 min. to 3min. will be consideredas metabolitesof D-1.
On the basis of the above results, it can be concludedthat sampleD-1 is not bioaccumulativein carp under the prescribedtest conditions.
Fig-I Infraredspectrumof D-1 (beforestartingthe study)
X: 4000.0.400.00 cm-1: 0.06.65.92 XT
4 scans: made ratio: resol 4.00 cm-1:
2812IG 0-1 Lot.No.101
threshold 3.00%: band
cm-1 2966.8 145a.6
% 46.13 46.61
cm-1 1730.3 1392.0
% 13.67 16.90
cm-1 1638.4 1202.5
1062.0 29.33 1017.7 28.37
984.6
808.0 35.17 706.4 44.98
764.1 48.20 653.5 33.43
746.7-
18 peaks found
92/05/20 11:00 apod strong
N.Saeki
% 46.05
6.36 27.24 44.54
cm-1
%
isis.a 53.2a
1148.9 7.71
857.9 S7.01
73S.7 45.63
65.92XT
Vc. C,
0.06 4000
3500
3000
2500
2000
1500
@le tooo
Fig-2 Infraredspectrumof D-1 (at the terminationof the study)
X: 4000.0.400.00 r-m-1: 0.23.76.30 %T
4 scans: tnode ratio: re5ol 4.00 cm-1:
2BI21G 0-1 Lot.No.101
threshold
3.00%: band
cm-1
3854.2
%
70.59
cm-1
3751.9
%
7 1.78
cm-i
3676.2.
3530.0
71.45
3587.7
71.30
2987.1
1638.4 60.89
1467.8 60.04
1392.9
1062.7- 46-04
10'18.3 44.44
985.2
784.3 59.95
746.9 56.19
735.7
19 peaks found
92/08/07
apod strong
%
71.50
64.35 31.11 42.65 56.80
cm-1
3650.1
1731.7 1202.3
808.2
09:57
x
71.29
29.76 23.91 49.13
ce" 500
76.30XT
Vc -1!
0.23 4000
3500
3000
2500
2000
-2R-
V40, V.10. vc:.
1500
1000
ce" -MO
Fig. 3 Bioconcentrationtest system
feed solution supply pump
p
dilutionwater supply pump
p
tap water
aeration
feed solution tank
0
0
activated
charcoal
(dechlorination)
dilution-watertank
2 5 2 OC
.......... .. .............. imixing tank
siphon
..........
...............................
...........V......:....:...:...................................
........................................
................................
..................................................................................
.....................................................................................................................................
..............................
.............................................
..........
glass aquarium 8 5 f (amount of water 8 0 .2
waste treatment
Fig-4 Gas chromatogramsof D-I(Standardsolution) for calibrationcurve
oggimi
(n-hexaneonly)
0.25gg/al
OJ @iD C3
I CD
0.50AS/111
r Lo c.;
N
1.00A
it
JD
U7
C-i
ce
-D
4m
'.0 LO -71
N
10
co
C4;
%z Im w
w
C-i
W"
co
to
U'p
-7. V)
<IC
.0
4x r
oc Im oc <K
Fig.5 Calibrationcurve of D-1
Y = 3.8338X 103 + = 0.99995
1.4473X 10.6 X Xl
x 1000000
1.6 1.5 1.4 1.3 1.2 -
cn >
0.9 -
Cd 0.8 0.7 -
,e 0.9 -
0.5 0.4 0.3 0. 2 0. 1 0.0
0.0
Calibration Curve
0.2
0.4
0.6
0.8
Concentration(g g/ml)
t
1 .0
1 .2
x1
START
Ptl%tio
1 2 3 4
START
@,4,65 67@1
T IliE
2.965 3.35 3 92 4.6 1
TOTAL
AREA IIK I BliO
2rj297 13721 v 628854 'V 137403 V
80,3275
colic
2.5362 1.7146 78.5797 17. 1695
11313
oq cCnD
mco
<
WARNING START
NO PEAK
< 30000 14ARMING NO PEAK
Fig.7-1 Gas chromatograms (Blanktestfrom fish) (beforestartingthe study) Std.O.50A &/aI
c@*) 41:0 tr) 0.11 0 (%.bl):
vI Cl)eo c@'-I
0 It
%1) OJ 1 0
to -n cel -7% 1 tr)
LLJ
ix
nr
v
C-J
C-J ce)
tf)
Nt -Di
07@
LLJ
to
0
u)
oi
v
v <I,
.C%O --N'w
V)6 Ir
w
OJ
0-
v
LLJ Le <r
ces-".rj%
LLJ
START
r4b cn
-55
r-n CD
P K li0
T I liE
AREA tiI-' I DTIO
colic
1 2 3 4
START
3.025
19079
3.413
12204 V
4. Ei1
639675 S'T'
4 . 72!5
139A 48 '4
---------
TOT-AL
810706
2.3534 1.5793 78. 9C,34 17. 1639 - ------------ -
11313
@4 4.205
CAD
p 11t%i0
1 a 3
START
T IliE
AREA MK
1.535
8144
1.928
35586
4. 2rj5
35EI 16.
---------
TOTAL
78746
I DHO
colic
10.3422 45.1906 44.4672 -------------
100
15-j4jZ
@ 4. 2j3ZJ
< 30000&V - Iec- -
PKIIO
1 2 3
TIliE
AREA MK
1.532 1.93 4. 2A2
TOTAL
7659 33256 26554 ---------
6747EI
lDhO
colic
11.3524 49.2902 39. 3f-74* -------------
Elri
r=+
CD
r+
CD O'Q
cCn+
En
CcDn e+
Table 1 Recovery test from water
w
A
B
C
amount of water
added amount
peak area V sec
ML 1 1000
Ag
Sample Std.
10
723856 762903
2 1 000
10
720337 762903
Concentration of standardsolution(Cstd.) Final volume (V)
Calculation
Cstd x B/C
E=DXV
D
conc. in final volume
E
measured amount
R
recovery Fig. No.
g 9/ML 0.474
A9 9.48
YO
94.8
8
0.472
9.44
94.4
8
Average
94.6 %
0.56 mg/mL .:20 mL
R=E/AXIOO
Table 2 Recovery test from fish
w
Fish weight
A added amount
B
c
peak area
V -sec
D
conc-in final volume
E measured
amount
9
IL9
sample
Std.
/Igimt
gg
1 36.@2 10
803266
813996
0.493
9.86
2 34.25 10
813149
813996
0.499
9.98
Concentrationof standard solution(CStd.) Final volume (V)
Average 0.5 gg/mL :20.OmL
R
recovery Fig. No.
YO
98.6
9
99.8
9
99.2 %
Calculation
D= Cstd x B/C
E=DXV
R=E/AXIOO
Table 3 Acute toxicitytest 48 hrs)
D-1 g g/ML
number of dead fish test fish
0 ( control
10
0
100
10
0-
mortality .% 0
0-
A.qhr-if.,tn= .sinn ticr/mL
ci SIART
PKIIQ
4. @,93
9@3
T I IIE
AREA
MK I DIIO
CONC
OR
CD
ri CD
3.315
13901 v
1.7455
rn
3
3.9
625788
V
78.5757
4
4.593
137115 v
- - -- - - -- -
17.2165 - - - - -- -- -- -- -
TOTAL
796415
100
co
CD
START
"%
P K ti;3
1 a 3 4
START
4.5-92
T I liE
AREA liI,' I 1)110
colic
2 . '7.4 4 2
16717
3.318
13213 V
3,898
59325e.. V
4. 59'2
130598
v
---------
TOTAL
753786
2.2177 1.7529 78.7037 17.3256 -------------
1013
p 11%110
2 3 4
4.595
3. 9k35
7 1 liE
AREA. IIK I B 110
ciliic
2.938
165,@5
3.318
12282 v
*3.9C'5
592950
v
.4.595
127387
v
---------
TOTAL
749214
2.215 1.6393 79.1429 17.0027 -------------
lf3l3
.START 0
35.567,
C/2
4-Lg@4.56?
FL CD
PI%,NO
T IIIE
AREA 11K 11)li0
cotic
CD
iz
C0D 0
CD
1
2.93
2C,325
2.3961
2
3. 3rj5
13939 V
1.6432
3
3.87-13
667862 V
78. 73@42
r+
4
4..567
146134 V
17.227.5
---------
-------------
0
TOTAL
8482613
1013
START
Ci. @3% 5@--4 .@-7-53
3...-85
p 11l%i0 1 2 3 4
START
T IliE
2.15 2.515, 2.932 3.885 4 .'Z7-
TOTAL
AREA liK
201E.6 6354
33246 V 650164 V 153102 y
863032.
I DtiO
colic
2.3366 k3.7362 3.8523 75.3348 17.74
-----------
C'
i 77''@
'SZ2--
4.583
p 11r%i0.
1 2 3 4 5
T I IIE
AREA MK
2.17@-.
15969
2.937
3'@54 3 5 '1
3.333
16576 V
114.893
657675 V
4,52.3
1!5547 4 y
---------
TOTAL
$78128
IDNO
colic
1.8185 3.6937 1.8876 74.8951 17.7051 -------------
1 ri0
CA
4 i.37
PKIIO
7 1IIE
AREA 111.' 1 DIIO
colic
IIAIIE
1
2. ?7
18144
2.2954
3.345
1 1535 v
I .465;,
3.
624824 V
79. A45 k
4
4.637
13511 1 1)
17. 1933 -------------
TOTAL
29rj465
lee
C+ 0
START
OFq
44 .(.44
3.-93j88
I IliE
AREA 11r, I DtiO
come
liAllE.
17 1;@'?
2
3.357
1 1133 7 14
3
3.938
551 147 V
4
4.642
13,@287 14
- ---------
TOTAL
714637
2.4@S 1.5416 77. 1ZZ6 13. 93e9 -------------
ISO
CD CD
START
312?
3.91
CD
l@@-4.
1,117
@n
TIIIE
AREA liK I DliO
colic
liAliE
ot
2 3 4
START
2.957
i.3.03
13763 v
3.91.
627773 V
4 . @rj7
136211
v
---------
TOTAL
798464
2.5946 1.7237
78.6223 17.0592 -------------
1 DO
92
44 .61177
-.-92
06
Pillit)
1 2 3 4
I liE
AREA liK
17479
3. 33@5
11633 V
@e
!.7;,722 V
13*882 V ---------
TOTAL
744716
Iblio
colic
2.3471 1.5621 77.4419 19.64:a9 -------------
1 ri
liAliE
Fig.11 Stabilityin water
100 80 Residual 60 D-1 (%) 40 20 01
0
'2 Time(days)
START
4 i,6L-
3.95$
CD
aq
T IIIE
AREA III,' I DIIO
colic
flAtIE
CIO
2
3.363
11934 V
3
3.958
6z5846 v
4
135512 v
---------
T Of AL
791374
I .5t)77 79.(4634 17. 1192 -------------
0 ri
CD
START
FL
3.
F,@46. F,6r9i
CL
PL,'HO
T I IIE
AREA lig IDMO
colic
IIAIIE
CD
1 2 3 4
START
2.99 3.36'8
16:476 11096 6(45190 v
4.668
144705 V
---------
TOTAL
779474
2.3763 1.4233 77.6418 18.5644 -------------
ISO
0
r.+ CD r+ cn c-
C:)
L
in_
963r
.667
CD
pklio
I IIIE
AREA [iK IDMO
colic
liAliE
ce
1 2 3 4
START
2.992 3.37 3.963 4.667
TOTAL
19238 13161 V 662222 V 14574CJ V
S41161
2.2871 1.6598 78.7271
17.326 -------------
ISO
f?@
rE, 4.641
3.95
pklio
1 2 3 4
T IItE
AREA lig
2.975 3.35 3.95 4.647
TOTAL
20201 12086 v 635744 V 158660. li ---------
818691
I DliO
colic
2.4675 1.4762 77.653? 18.4025 -------------
1013
liAllE
Fig-13 Stabilityin feedsolution
100
80 Residual 60 D-1
40
20
0
2
4
6
8
10
12
14
Time(days)
Table 4 Analysisof D-1 in testwater (Highexposurelevel)
sampling date
i
exposure period
A
B
peak area
g V - sec
c
conc. in water
E
mean conc.
Fig. No.
day week
Sample Std. gg/mL 'ag/mL
5.26, 192 0 0 1 691668 761885 0.096 0.096 1-1
29
3
2 1551911 768251 .0.076 . 0.086 2
6. 2
7
1
3 665450 759i99 0.093 0.088 3
5
10
4 684891 764544 0.095 0.090 4 1
9
14 2 5 655152 752597 0.092 0.090 5
12
17
6 742762 794882 0.099 0.092 6
16
21 3 7 600994 794596 0.080 0.0901 7
1
19
24
8 1 705686 824875 0.090 0.090 8.
23
28 1 4 9 1716171 792816 0.095 0.091 9
26
31
10 1687002. 809639 0.090 0..091 10
30
35
5 11 1776203 831148 0.099 0.091 11
7. 3
38
12 1709193 787890 0.095 0.092 12
. 7
42 6 13 674771 77106@5 0.093 0.092 13
10
45
14 742351 1808504 0.097 0.092 14
14
49 7 15 701150 1791505 0.094 0.092 15
17
52
16 544487 775383 0.074 0.091 16
21
56 8 17 733512 1779453 0.099 0.092 17
Concentrationof standardsolution Final volume Sample volume Recovery
Cstd V:
w: R:
0.5 20 100 94.6
*
g g/mL mL ML %
Appendix-2
Calculation
C = Cstd x A/B x F
(F= V/W/ (R/ 1 0 0
E = E C i/ i (i: numberof analysis)
Mean concentrationin water -s-tand-a-r..d-.d.ev-:..ia6t-i.o0n92 0.0677 @Tg7mL
Table 5 Analysisof D-1 in test water (Lowexposurelevel)
sampling date
i
exposure period
A
B
peak area
,aV -sec
C
E
conc. mean Fig.
in
conc. No.
water
day week
Sample Std. 9 g/mL g g/mL
51.26, 192 0 0 1 738542 .761885 0.0102 0.0102 1-1
29
3
2 359497 768251 0.0049 0.0076 2
6. 2
7
1
3 515797 759199 0.0072 0.0047 3
5
10
4 599070 764544 0.0083 0.0077 4
9
14 2 5 570779 752597 0.0080 0.0077 5
12
17
6 572050 794882 0.0076 0.0077 6
16
21 1 3
7 659682 794596 0.0088 0.0079
7 1
19
24
8 572309 824875 0.0073 0.0078 8
23
28 4 9 676137 792816 0.0090 0.0079 9
26
31
10 689946 809639 0.0090 0.0080 10
30
35
5 11 701085 831148 0.0089 0.0081 11
7. 3
38
12 636904 787890 0.0085 0.0081 12
.7
42
6 13 615794 771065 0.0084 0.0082 13.
10
45
14 662294 808504 0.0087 0.0082 14
14 49 7 15 606422 1 791505 0.0081 0.0082 15
17
52
16 591058 775383 0.0081 0.0082 16
21
56 8 17 600028 779453 0.0081 0.0082 17
Concentrationof standardsolution Final volume Sample volume Recovery
Cstd V:
W: R:
( * Appendix-2
0.5 g g/ML 20 mL 1000 ML 94.6 %
Calculation
C= Cstd XA/BXF
(F= V/W/ (R/1 0 0)
E E C i i (I: numberof analysis)
Mean concentration In water 8@tandgtd'deViation 0:0082 O.CGII*Ing-7mL
Fig.14 Concentrationsof test substancein water High exposure level
0.20 0.18
0.16
Nominal value
Analyzed value
0.14
0.12
Concentration 0.10L@ .............................................................................................................................
u g/mL) 0.08
0.06
0.04
0.02
0.00-
0
7
14
21
28
35
42
49
56
Exposure period
(days)
Fig-15 Concentrationsof testsubstancein water' Low exsposure level
0.018 -
0.016-
0.014
...Nominal value Analyzed value
0.012
Concentration 0.010 (/ig/mL) 0.008-
.......
0.006-
0.004
0.002
0.000 0
7
14
21
28
35
42
49
56
Exposure period .(days)
Table 6 Analysis of D-1 in test fish (Highexposure level)
w
A
c
D
exposure fish period weight
peak area V -sec
conc. in conc.
final in fish volume measured
week
2 .1
2.
4
1
2
9 34.14
Sample 90324
Std. 839568
34-80 94243 796908
37.18 168720 807176
39-96 192918 790270
g g/mL 0.054 0.059 0.105 0.122
g g/g 0.032 0.034 0.057 0.062
.6
1 36-95 161768 839289 0.096
0.052
37.47 113668 804605 0.071
0.038
8
1 35-54 132230 879847 0.075
0.043
2 36.77 145406 885954 0.082
0.045
E
F
mean conc.
BCF Fig. No..
in
water
g g/mL
0.090 0.4 18-1
0..090 0.4 18-1
0.091 0.6 18-2
0.091 0.7 18-2
0.092 0.6 18-3
0.092 0.4 18-3
0.092 0.5 18-4
0.092 0.5 18-4
Concentration of standardsolution(Std.) Cstd
Final volume
V
Recovery
R
0.5,ug/mL 20 ML . 99.2 %
Calculation C =Cstdx A/ B
D=CXV/WxlOO
/R
FDE
Table 7 Analysisof D-1 in test fish (Lowexposurelevel)
w
A
c
D
EF
exposure fish lperiod weight
peak area V - sec
conc. in conc. mean BCF Fig.
final in fish conc.
No.
volume measured in
water
week
9 Sample Std. g 9/ML
g 9/9 g g/ML
2
1 34.73 72837 839568 0.043 0.025 0.0077 3 18-5
2 35.61 61435 796908 0.039 0.022 0.9077 3 18-5
4
1 34.23 93655 807176 0.058 0.034 0.0079 4 18-6
2 35.70 63936 790270 0.040
0.023 0.0079 .
3 18-6
.6 1 1 37.23 38016 1'804605 0.024 0.013 0.0082 2 18-7
2 1 40-00 53795 798557
0.034 0.017 0.0082 1
2 18-7
8
1 35.27 45555 851984 1-
0.027
0.015 I 0.0082 I 2 1 18-8 1
2 35.73 1 46433 1 864383 1 0.027 0.015 0.0082 2- 18-8
Concentrationof standardsolution(Std.) Cstd : 0.5gg/mL
Final volume
V : 20 mL.
Recovery
R : .99.2 %
Calculation C = Cstd x A B
D=CXV/WxlOO
/R
V=D/E
Fig.16 Bioconcentrationfactors (BCFS)of D-1 High exposure level
................................................................................................................................................................................. .......................................................................................................... ........................................................................................................... ........................................................................................
........................................................................................
BCF
....................................................................................................................................................:........................................
........................................................................................................
.........................................................................@.@o.............................
................................... ...................................................................................
.......................................................................................
.............................................................................................
2
4
6
8
Exposureperiod (weeks)
Fig.17 Bioconcentrationfactors(BCFS)of D-1 Low exsposure leveI
10
.....................
............
..............................................................................................................................................................................
.......................................................................................
..................................................................................... .............................
..................0....................................................................................................
.....................................................0...........................................................................
BCF
1
................................... .................... ........ .......................... .........................................................................
......................................................................................
...................................................................................................................
.................................................................................................................................
......................................................................................
......................................................................................
0. 1
0
2
4
6
Exposure period (weeks)
START
pi.lia 1 :3 4
START
47.6;
7 1 IIE
ARECT
119 1 DIIO
2.376 3,$73
T(ITAL
14921 '4 66as4i V 15(07 11 v
---------
8763$;*
colic
liAllE
1 .7CJ26
79.4009
lip??
----
Pi.ito
3 4 5
START
313 i0.,@i37)S
43 ppis
T IiiE
AREA IIK Iblio
colic
22:3 2.$2 3.313 3.9.)7 4. @$ 4.a3
iiesi 17;339, 1?7252 v L?45,i 4:3911 v 184@77 v
S. 165
62e74 v
------r--
TOTAL
96S141
1.746 64.0133 IS. 3,;54
2.8449 4.559 1. 1551 6.3145 -------------
I coo
itAltE
PKIIO
1 2 3 4
START
4L64?.3673
7 1tiE
AREA lilt lblio
colic
2. i?97
le$95
3.373
11236 v
3.9ce
65se7k v
4.673
141,331 V
---------
70TAL
626746
2.2492 1.3391 79.3329 17. ?3397 -------------
1e,3
lialic
PKITO
1 41, 3 4
5
6
a
3.365
2.01
154@ 30
I IitE
AREA
iii, Ibilo
colic
2.036
6617
2.2(07 2. i;I 3.303
3.
4.333
S363 46?t$4 lSi763 V
2of
@5f$54 V
4.775
1?7(47 v
1..14
34727 -4
---------
70TAL
77 1
6.6577 1.213i Le.ioa? ZO.3113
6ee?
7.23$6 2.21)51 4.SCJ12 -------------
II)a
[tAllE
C:) tn CD
00
En
OQ
OD
t
0
CD OIQ
co
0in
a
En
CD
r_
(D
CD
CD
>-
<
CD
02
cn 0
CD
cn
9=L
cn 'f--
CD CD
4 . 4a
Pi.ilo
4 1 1iic
AREA 111' 1 blio
colic
IIAIIE
START
3.367 3, @-IB 4.i$
TI)TAL
13694 V i 623 1 14 4 ---------
94 I'@Il'I
3. !kL-
-------------
3.29
2.71
PKIIO
1 2 3 4
6 7 8 9 10 11 12
START
I ItiE
AREA 11 l@
1.657
305$i@
1.21 i.fil$
6!.Z,6677 194391
49740 V
?.348 79
3.2a3
53@16 3i.284 iolii&S
3.1$65
4m2?o
1 14 Fzg-a .4
4, ?7e:
3521@, *,p
5.133
4 3:ii?- -.1
---------
TOTAL
Z614127
I DIIO
colic
23.9654 ?..1339 I.Cie2i 6.2(134 1.3fs?6
42.C4534 13.?513
193 4.3663 1.34i&6 1 .;a -------------
lei)
IIAIIE
PKITO 1 2 3 4
START
Pxito
2 3 4 5 7
I ri 11
4. ;.*:?
Tinc
AREA iiK Iblio
colic
litille
3.(400 3.392 3.98i 4.667
105!li@ 13323 v 647973 V 14LIt?_ V
707AL ---e-2-i-i-5-e-
2.257 1.6205 70.8145 i?.3as -------------
loo
3 45
7 111E
AREA IIK
33016
1.295 1.1;1? 2;Cil * 2.1@7 2.717 3.292
97il 27*
671331 l9?s96
50049 V 125F;i. 93666?1 361&954 v 4k27,& iii9il. V
4.763
3ei47 v
5.147
i.?i&l v
---------
7CiTAL 246cl 1;@1
1.295 2.797
Iblio
colic
IIAIIE
1.3371 27.31)09
9.6147 2.0593
37.61)4 12.4214
i.71ka 4.8@97 1.2412 E.?524 -------------
166
00 a
:z 0 CD
(D ;q
co
SD En
0 a
aq
OD
CD Uq
x
13
Ila co
0
E3
cn (A
co
CD
<
CD
gn
tn 0
CD cn
cn
CA
CD @n CD
CD CD ;q
si Lik 7
2 3 4
SIAR7
7 1tiE
AREA 1%9 1
3. Ct? 3.463 4-67,%
Io)7AL
a ib 13911 '4 @*1 la v 14989? v
-------7-
fa?4 ZOJ
colic
IIAIIE
2!.40 17
1.5$21 ?6.*593 1?. 1467
-------------
4
5
-i
7 a
10
10 I 12
s7;4A7
4.11 Sg 3. 4@53
I ISIE
AREA III,* Ibito
colic
i.;1*3 1.32
1 1 .9,) @,
z.233
@@. 633
2.93$
3.453 4 . C4.1@"
707AL
5143* 1 Ci4 C$3 '4
1 -&4 2')!$ V
34433 V
3404*
. i-@27733 Y
a3i,23 1
15450
1 ie*Z V
34ZL".:: v
-------
14 -
2796831
1.9378 34. 31 @:i
(47;7
5.37
1.2382
1.2451
36.7195
9.4402
6. 3.19962 1.2372
2. 486')
-------------
lot)
titlile
pxlto
3 4
s7AR7
PHHO 1 2 a 4 7
11)
4L;4a. *22;;@t;
4.107
I IiiE
AREA IIX IDIIO
Colic
3. 4 9;3
17*20 13091 V
4.1(47
659094 v
4.039
145511 v
---------
707AL
$33565
2.1328 1.5656 73.9637 17.4159 -------------
100
itAnE
4. 4z.
7 111E
b@83
3.397 4-riii
AREA
IIK
1.1
103145
1.333
]($212ai 4
l.tt3
171all-:
2.C$75
3$347 v
2.262
9 ?1? 1)
e.ele
31831
2.683
e&Zi,(03 v
3.397
3t3l53 v
4
4. 4 )!a
e9i55 .4
4.945
14 C49i: v
5.283
52icie 14
---------
707,IL
2.737177
2.993 1.332
Ibito
colic
IIAIIE
3.7683 37.664
i,.2470 1.4448 6.@L46 l.li47 31.5143 it.6061 6.3771 3.2iO3
(J.5146
1.1327 -------------
loo
CD
CD
'I--
Z?
aq
00
cn
0
(xD 0
10 0
CD
CD
cn
CD
CD
<
-
CD
rn
0
CD
En
r+
cn
cn FL
F
cn cz 'I--
:E CD CD
Fig.18-4 Gas chromatograms (Analysisof testfish) High exposure level
Std. 0.50,u g/m I
week 8 No. 1
Std-O-SOg g/ml
v r, -D 1 -)3 -n -o n ii
VI .1
V't -.V
n
W)
,44
4w III".
a :a v 1 4,j - N
4n No
ti I V
:"Jj
ij
C2 Oj
week No.
7 1liE
,RE.; II,I' IUIIO
4 START
TOTAL
14121 OL
S.@@7 V? v ---------
colic
IIAIIE
17 i,@77 t-oo-) ----
Piltio
Z.033
2.317 33..137137 F 445 4??Is
173
I liie
I bilo
colic
11.111C
2
174 93,L
3
3.317
:39lz V
4
4. GI)CI
;3412
5
4.4a5
644*@k5 V
3.173
220 73 v
---------
31.7735 13.iF35? 2. 4fj'9i, I?j.Oii7i
8.0125 -------------
START
PKIIO
3 4
START
34!.6373
3. :Pils
I Ilic
AREA
III' I
1*505
3.373 3.9i.4
1123 & V
655*77
v
4.,&73
1411)31 V
---------
92J,740
colic
2. 24-?Z 1.35,91 79.3329 1?.6387 ------------lee
iiAiiE
PKIII) 1
4 6
7 12
S. 17is
I liie
AREA
IIK
2:.S;,7
7704
jZ631$ v
379Le
V
9787 v
1583(.
4 .38:3
45571
'4
3597(s ---------
TCJTAL
363;141
I Dito
colic
2.3412 49.3667 12.3117
3.2265 5.2385 i3.Ci355 II .*i,59 -------------
Sao
is.;;IE
CD
oq
00
cn
cn
0
r+
(lq
0 E3
cn
CD
m
co
CD
CD
cn
0
CD En
r+
cn CD
00
CD
CD
Fig.18-6 Gas chromatograms (Analysisof testfish) LA)w exposurelevel-
Std.0.50,ug/ml
week 4 No. I
5 td.0.50,u g/m I
week
No.
.0OJ 'n
Ix 42
-M I @j I
43,
-Z .0
Y. r..Ai 0
K
ti
09
OJ 0 -T
Ix
Iz
%9 dL
Ui
I-)
-Z
V
lot. 'V4
Li % Iz
el
-Z
dL
44
SIAR7
pxfio
4 SIAR7
2 3 4 5 1& 7 0 9 10
SIARI
4@84 32;S11J
I InE
AREil liK Ibigo
colic
ley
17@@Zfi
3.493
131)61 14
4. 167
v
$30 ____L4 1',.I-L V ---------
IOTAL
$)355CJ5
2. 13k;?
1.5656 78.8037
17 4159 --------*-----
lie4liE
6 7" 1.322
3.4 4 )4 57
5.313
7 1IIE
2.69 AREA 119
1 DIIO
colic
ItAllE
1.(487 1.022 1.05 2.0?3 2.633 2 . C$9 3.4
173.%4 286123
15659 v 6451
225211 601 1!5 v
4.1 4,457 5.315
a 1 C457 v 4 (i3.4 3 ---------
731060
2.3713 39.1411;
5 4 7*.l 2.1694
st-i!5 30.88)06
9.3161 0.952 4.2406 5.56136 -------------
1 lae
1.0(.3
3 4
SIAR7
4.39'CS3C3
4.147
I IIIE
AREA ltx I DIIO
colic
3.135 3.z2 4-14Z
707AL
16317 11903 14 L5497ft V
v ---------
024056
1.9734 1.4492 79.@1003 17. 3?;p 1 -------------
Soo
ilAtIE
Pilito
a 3 4
7
le
3.42f$ 44@7
S.3(.3
I IIE
2.917 AREA ISK
1.678 1.323
406(49 296921
1.867 2.092 2.638 2.917 3.42$ 4.167 4 '1 5.363
42323 55449 V 13416 289337 V 93474
6361 4*1.'44 V 36CJ14 ---------
ICJIAL 520567
I DIIO
Colic
IIAIIE
4.346 32.2542
4 . 651 (t 6.1)233 1 .4 71:1p 31.3272 IH.154
0.7433 5.01)63
------------IGO
CA
c.n c@l
cm
00
0
0 CD CD ;q
C)
Do C+ 0 CD O'Q >4
's Ein3
CD
CD >.
<
m
CD go
0 CD
C/3 cCL
CD
s
:z :c
0.
CD CD
;q
Fig-18-8 Gas chromatograms (Analysiosf testfish) Low exposure level
Std.0.50.ug/mi
week 8 No. 1
Std.0.50,ug/mi
week 8 No. 2
li,i cu
Iz Ix 43
Li
I:r
ag
19 Iz
lj P.
b)
*1
oa
,
Ai
cu -j o
tJ
uo
X
-Z
.10
tki
Ai
1)
ca"rlti
fin
ID e xI
Infraredspectrumof D-1 Data provided by the sponsor page in all
76.760 PEAK XT
THRESHOLD cm-1
0.73 XT rr
69.444 3452.0
0
2986.00 52.995
2-361.00 70.795
62.129
1940.00 1734.00
75.420 12 265
i63S.OO 54.*037
ISM.00 62.462
54.ei2 - 1537.00 1464.00
72.785 52.695
1446.00 !3.337
1397.00 8.690
47.496 - 1353.00 ew;,S 1296.00
31.947 17.422
1245.00 4.305
1207.00 3.600
40.180
1109.00 11.632
1061.00 27.475
1045.00 14.660
32.864
1034.00 9M.00
13.040 26.982
971.00 34.585.
SZ7.00 30. EF75
905.00 25.!)48
.974.00
14.750 32.215
810.00
9.475
747.00 49.700
IS pliA;,Rr-6.00 45.812
726.00 49.120
707.00 46.817
654.00 31.735
io.sis .588.00 38.612
554.00 30.755
5.3032842.00 34.780
55r4@064.410..-0004 46.350
4000
0 67.110 3600
2900
2000
isoo
1200
Boo
cm-1
-Ott
'7
RE
400
A 1D iD e xi
i :@c 2
Gas chromatograms(Analysisof test water)
17 pages in all
START
crq
CA
4.6138
CD
p 1%N, 0
T I IlE
AREA tl 1'% 1 DIIO
colic
@n
n
CD
m
I--
cn
04
a
0
2.95
1*728
a
gn
3.327
12342 V
1.5564
r+ 0
3
3.913
626699 v
79.0334
aq -1
4
4.6138
135186- V
17.0484
co
---------
-------------
TOTAL
792955
START
0
p 111%10
4.*6 1
.3.917
TIIIE
AREA IIK I DliO
colic
CD
cn
CD
r+
>C
CD
1
2.95
17757
2.458
CD
2
3.327
12979 V
1.7967
3
3.917
56CI655
77.6097
CD
4
4.61
131013
v
---------
18.1357 -------------
TOTAL
7224k33
lee
START
3.918
0
4@6.1 7@
T-
CD
x
p 11l%i0
T I liE
AREA ti11% 1 DtfO
cotic
cn c
13
CD
1
2. 953
1.-".199
2. ':.i.72
2
3 35
1 E. 1
1.5688
CD
3.918
79.2617
4
4.i:,I?
129177
v
---------
16.8023 -------------
TOTAL
7 6 8 *,1D32
START
4.533 TOP
l@IN,lii.'i
7 1 IIE
1
2.9
AREA 18925
111%' 1 DliO
ciiiie 2.3642
4 START
4.533
136615
v
---------
TOTAL
800456
-------------
2 3 4
.START
457
T 111 E
AREA
11
2.925
12;3cl 1
3.295 E,7 5
9318 v
443612
Y
4.514,7
lrj829,9 v
---------
TOTAL
574CJ3,3
1 DIIO
1.6232 77 . Ee.C,3 18.8665 ------------1130
cn
c.n CD aq
CcLo C.0
0
cn
0 e+ CD cn r+
CD
C+ CD
x
0 cn
(D
3.873
4.557 0
5 . 5 4;4525 7
:c
CD
x
T I IlE
AREA
111'% IDNO
colic
2
-S,
9 Cl9 4
3.292
5 6 1 ;,:
3
.29 1 C,49
4
-4 - cj57
68448
'4
5
5.542
17797 Y
2.32
CD
1.4326
? 4 .2 4 63
17 .4 n'1
4.54Cl
-------------
TOTAL
----3 -9 -2 -0-Ce-4
1
TA R T
a 3 4
START
4.577
3.:3 9
T IliE
AREA
2.
3. 3ris
13423 V
3. z@:9
625413
V
4,577
133786
Y
---------
TOTAL
791934
IDNO
colic
2.438 1.6956 78.9728 16.3935 -------------
10
Pll%tio
1 2 3 4
START
T IIIE
AREA
111'% 1 blio
colic
2.942
1@!,715
3.317
11039 V
3.9
537225
Y
.4.592
128225 v
---------
TOTAL
692239
2.2754
1.5946 77. 6C[68
13.5233 -------------
Oq cn
CD I-- im =r em "S
0 a 0 orq
3
rn 0 r+ (D aq r+ CD CD 0 CD CD
3.898
0
ze
4.955@955
CD
0
pklio
T IliE
AREA 111%' 1 BliO
colic
1
2.942
iiess
2
3. 3 1'
8453 V
3
2;. 8191.3
412616
Y
CD
2.IE17? CD
1.5734 @g 77.047
4
4.59.5
1rj31;81 '4
19.2669 -------------
TOTAL
5 3 5 5 3.3
100
START
PKIIO
1
2 3 4
STAR*T
4.55@7l3
TIHE
AREA
tl1'% 1 DtiO
c 0 11ill
r@r.r1i2
13275 V
628548 Y
4 c;73
135996 '4
---------
TOTAL
797820
2. 5C'71 I .r,639
78.7831 17.0459 -------------
i=-4
i: '46*6-
4.5757@5,5
3.883
PKIIO
1
2 3 4
START
7 1IIE
AREA
111.' 1 DtiO
cotic
2,933
133EI8
3.302
12432
Y
3. 8 8 ZI.
551308
IT,
4.57@4
133583
V
---------
TOTAL
715630
2.10-583
1.7371 77.038 18.6665 -------------
IGO
00 CD
En
CD fn
r-t
aq
ze
PO
C-+
(D
CD
x
cn
CD
CD CD
CD
0
PKIIO
T I liE
AREA
tiK 11) 110
cotic
CD
1
2
2
3.3(13
10755 V
a
3. 88@..
479,@51
y
2.4128
1.721
CD
76.8042
4
4.575
119119
14
---------
19. ei:12 -------------
TOTAL
6249i33
lee
TA R T
PKIIO 1
a
4
START
4
T I IIE
AREA 111'% 1 D 11 ri
colic
2.95 3.318 3. 9E17 4.598
TOTAL
19573 12944 V 6 1* 1(15* Y 134492 V
---------
785115
2. 4?3 1 1 .64-37 73. 72e. 17.1300@
-------------
1,3ri
(17 4.660055
PKIii3
1
2 3 4
START
T.1 11E
A R E A. 111'% IDNO
c 0 ti c
2.955 3.33
16 3 1 1i;52 v
@-:.9El7
526234@- V
4.605
128918
v
---------
TOTAL
683437
2 . 4 3'3 1.7049 76.9982 13.8630@ -------------
CL
@D
61 CD
tw
iz
CL
00
m
L<
0 03
3
cn 0 e+
CD CD
En (D
CD
CD
325
W-
9C-13 CD
44 . 6P;
cn
c
p 11%110
T Ih E
AREA
111'% ID"O
CONC
s
CD
I
a. 9!f.
2
. IS-325
14 7,;' 3 11091 v
2.4802 CD 1.8583
0
459129
V
76.9487
4
4.
11 16-513 '.1
---------
18.7123 -------------
TOTAL
596669
lee
START
c.n CD
T I tiE
AREA
1111, 1 Dtio
colic
2. 9E.3
2.2682
0
3.363 13121 V 1.537 =r
0
3
3. 9 5
651281
v
78.7756
a
4
4.9,65
143601
V
----------
1?.3692 -------------
TOTAL
826754
0 ri
START
p I,t,i0
1 a 3 4
START
44 .663
3.957
T I liE
AREA
MK
2.987
17961
3.36
12491 V
3.957
5 9 6 6 62 V
4.66.3
146100
V
---------
TOTAL
773214
I DliO
colic
2. 3221? 1.615--1 77. 1664
-------------
0
:r
CD
CD
C-+
CD CD
CD CD
3. 9E.5 0
p 11l%i0
T 111 E
AREA M 1'% IDNO
colic
1
2.997
12765
E. 14 4
CD
2
3.382
9--148 v
1.5733
3
9 @,5
459404
Y
77.3195
CD-
4
4.665
112646
V
-------
13.9588 -----------
TOTAL
594163
START START
cIn-
FL 3.95
652
C=)
Pil%ilo
T I liE
AREA [iK IBliO
c 0 11.11-
C)
1
2 . 9 *.2*,
18998
2.297.5
0
2
3.36
13311
1.6098
3
3.95
651927
73.8394
4 4.652 1-4-2-6-69-- Y ------1-7-.-25-3-3-- a0q
TOTAL
8269,35
11313
START
cn
4@6.5
Ct) CcDo C-+
T 111E
AREA liK I B li0
colic
CD
OfD-t.
CD
2. 9:3
2
:3.@53
1453.7 10426 V
2.3224
C<D
CD
I . @1656
3
@-:.95
4834*.;,1 'T'
4
4.65
117513 '4
---------
77.2387
IS.?733 -------------
TOTA.L
.625957
1ori
START
07r
0
957
CD
56@5
p I,t,i0
T I IIE
AREA IIK I])NO
colic
CD
1
2.985
155 1
2. 26CI9
(D
a
3.367
1 1 lefi 14
1 .it,289
3.957
529472 V
77. 1397
4
4.655
136210 v
---------
18.9765 -------------
TOTAL
686380
START
p 11%110
4. 602
T I liE
AREA li1'% 1 DIIO
colic
cn
c@
CD
2.952
2 93
2. 4 3CIA
3.332
13323 V
3.913
6751 16 Y
7 8 5":.s7
0
4
4.602
14 9 7 @-9 V
---------
17.422 --------------
TOTAL
859596
lea
START
2C 2.252
3,5" 6@4l.:--
3 92 8
C-+
CD rn
CD >C
0
T I liE
AREA
IIK I DIIO
colic
rn
r+
c
CD
CD
2. "97 3
19175
2. 6EI38
CD
a
3.353
11557 V
1.5693
CD
11:
3.928
567817
Y
77.1053
4
4.623
137869 v
---------
18.7211, -------------
TOTAL
73@,4.1?
1 ri0
START
4L64.&.69227
CD
cn
P.I'1%10
T I tiE
AREA
111-' 1 DliO
colic
CD
2.965
1;36;17
2.2947
CD
2
3.34
9585 V
I . 6rj94
CD
114
41..927
457676 Y
76.P,4e
4
4.62:7
114633 V
----------
19.2479 -------------
TOTAL
595561
iriri
T A R-1
PKIIO
1
2 3 4
START
4
7 IIIE
AREA [it'% IDtiO
colic
3
21055
3.293
14 -S 14
3. e.?3
6487(45
4.56
14411 1 V ---------
TOTAL
8281413
2. f-425 1.7229 -78.3328 17'.4 1118 -------------
p V%li0
2 3 4
START
@:7@6 11:1@
T ItlE
2.9513.327 3.91 4. 6r-43
TOTAL
AREA
1:?7 4 12935 575386 140785
747850
M 1'% 1 DliO
colic
2. 5cl.62
v
1./-297
v
76.9388
V
18.8253
-------------
p 11%11a
2 3 4
238 4 . I.IJ
3.913
T I IIE
AREA 111'% 1 DtiO
colic
2.963
1 e-682
3.347
25742 V
:S. 9 1 @-. . 54*.:-.C,95 Y
4.608
128042
V
---------
TOTAL
?213560
2 .59 3 .57'@J4 76.0652 17.?697 -------------
1 Cie
cn e-
CD n cn =r
-3 0
00 0
I= cn
r+ CcDn 90
CD CD
1x10 0 CA
CD CD
<
0
x 1!0 0 rn
CD
CD
START
p 11l%i0
1
2
4
START
963 44 .@667777
T 111E
AREA
9 cc
3.375
:3.9 6 4.677
19746
13661
662568
147071
---------
TOTAL
843045
ti1'% 1 DliO
c lic
2. 3422
V
1 .62,34
v
78.5922
%1
17@4452
-------------
ll3k3
P K li0
1 a I
4
START
4.66.5
T I liE
AREA
li1'% IDNO
colic
2.993
1671-13
3.375
1-1 146 '4
3.963
552061--1 IV,
4.665
134942
V
---------
TOTAL
714352
2.3366 1.5593 77.2272 18.8769 -------------
irie
Rq
C')
CD
CD
'@l-
cn
0 =r
CL
0
CIO 0
0
r+
CD
aq
cn r+
(D
x
>4
PD r+
CD
CD
CD CD
3.967
0
:c
4. iL;,5
CD
PKIIQ
I tiE
AREA
ti 1'% 1 DliO
colic
CD
1
2. 99;?
17@z"7
3.333
12302 V
3
9 67
552129
2.3687
CD
1.7103 9 76.7607
4
4 . iL65
137817
V
---------
19. 1603 -------------
TI)TAL
719286
lee
START
p 11%ti0 START
-4L
4 . 6 1 C-:
3.'9 2 3
7 1 liE
AREA
liK I DtiO
c iic
2.
Cl4
3. 3 4,'
13 14ri '-i
6 7 8 @'e2'.":
4.618
15282ii -- -- - - - - -
TOTAL
864r39l
I . 52ii? 78.5019 1 6356 - - - - - - -- - - - - -
1 ljl3
.225
3.92
p I,%1*10 1 2 4
START
T I tiE
AREA
ti1'% 1 DIIO
colic
2. '@67
21191
3.342
1437.3 V
3.92
625275
y
4.613
150928
v
---------
TriTAL
a 1 1 ?72
2. 6 1 C,4 1.7712 77.C,26 1:3. 59 2 4 -------------
1130
1.222 2C- 2.2E3'3:3@3
4. 618
3.'923
0
T IIlE
AREA lit'% IDNO
colic
E:.957
1 49
2
3.35
1 124Ci
2
569 17'@ Y
4
4 . ;,1
1:319 0 6 v
- ---------
TOTAL
728974
2. 2e.39 1.5419 78. '-:1794 18.0943 -------------
CD
C)
m
I
CL
m
0
CIO cn
r+ 0
crq
E3
cn
0
CD
CD
cn
CD CD CD
0 me CD x
Cl) CD (D
START
2 3 4 START
p 111%10 2 3 4
START
3.-972 6@j
7 1IIE
AREA 111'% 1 DtiO
colic
1 -'6*4 9
3.373
13443 V
972
A-4 3 7 @,.7 V
4.633
144133 V
---------
TOTAL
819982
2. 274Z's
1.6401
78.
1
17.57?5 -------------
1.298 5783 :3
-2 449.768
4. 057
T I IlE
AREA tl1'% 1 DNO
colic
Cl7
2lj3cl5
3.453
17773 Y
4.0,97
5696 1 A.
4.768
139577 V
---------
TOTAL
7472-71
2. 7 1:,3
2.3784 76.2262 18.6732 -------------
CL
CD
cm
cn
0 00 0
cn
r+
(cDn
=r
r+
CD
x lw
r+
0
(D
CD
CD
3.973
0
CD x 1010
PKIIO
T IIIE
AREA 11K I TJNO
coiiil@
2.'995
2
3.1@178
13573 10671 V
CD
E.C'53
CD
1.614
<
3
z". '?7:@
4 7? 123 'le'
72. 4681.1@
4
4.638
157779 V
---------
23.8644 -------------
TOTAL
661148
irie
r@T A R T
p 11%[113
3 4 START
44
T I IlE
AREA li1'% 1 DNri
colic
2.973
1 73
3.35
13235 v
3. 93@.
633159
4.64
1's? 9 116 V ---------
TOTAL
8rj3373
2.3741 1.6475 7;3. E.'l2 6
-------------
CS
4@ll4.65757
PKIIO
2 3 4
START
T I tiE
AREA 11K I DIIO
colic
2.987
16879
3.382
1263@, v
9 5 e.
5@'j9;7-'4ctJRC; Y
4 . f-5;7
135321
v
---------
TOTAL
7 0 4 3*37
2 1 . 1312 76.5898 19. a 12 -------------
11313
1. 132
'3 4.7275,77
4 . C"42
P.il1%10
1
2 3 4
T 111 E
AREA M 1'% 1 Dtiri
colic
3 . ci
14151
18364 V
4.r,42
494294
V
4.757
1 "115 clci IV,
---------
TOTAL
6 41-:1r3ic;
2.21 i.iLIS6 77.1962 13.9752 -------------
I
cn
CL
co C."
ECD
i3
C0L3 0
cn
CcDn
co
CD
CD
CD x
CD
9
S.TA R T
p II1I10
1
2 3 4
START
:1@42 3
4. 103
TIME
AREA
11K
3 . ciI?el
1 92CI2
3.488
12900 V.
4 . I El
9-.63595 V
4.823
144909
V
- -- - - -- - -
TOTAL
8 4 ri6136
I DliO
rolic
2.2843 1.5346 78.9425 17 .*-3:,a 7 - - - -- - - - -- - --
1 riL3
El
11,5 -c@
4 .SE:5
4.097
PKIIO
3 4 START
TIME
AREA 111' 1 DNO
colic
2
18955
12769 V
4. G@:07
594606
V
4.825
147745
V
---------
TOTAL
774075
2.4487 I .64 9 7i:,.:315 19.0867 -------------
1 ti13
C,..I1l.05577.
7-
4@8
18
4.1387
PKIIO
2 3 4
T I ME
AREA
MK
7
1 555
3.5133
31394 V
4.C,-.-7
537210
V
4.818
125084
V
---------
TOTAL
713742
IDNO
colic
2 . 7 *?8 4. 4635 75.2666
17.525 -------------
FL
!:> cn C:),
13
co
C/)
cn 0
CD
CD
cn
0
CD
CD
x
co
r+
cn CD
CD
CD
CD
CD
0 ze CD >4 Ila 0
CD CD CD
START
I rj6622 ."47
j4 4 7
4. 115
PKIIU
1
2 3 4
START
T I IIE
AREA 11 IN' I DliO
colic
:3.1 15
3 50'3
15023 V
4 1 I!T-
64531 1 V
4.847
146194
V
---------
TOTAL
32461'c,@ai
E. 1992 1.8218 78. C--513 17i?2?7 -------------
100
1.1362 2C-' 2.33"33"32
_@@4@3
42@
4. 11
PK[io
1 2 3 4
START
TIHE
3. 1 C'5 3.502 4. 11 4.842
TOTAL
AREA 11K
I 5C[6i.:. 10678 v 56626.3 'V' 134887- V
7 2 6;39 3
I T)ti0
colic
2.0726 1.469 7 7. 9 Fj 18 18.5566 -------------
1
p 11%110
.1 2 li 4
2.325 4L@..4. @.4.4@@ll
4. 1 1 @o
1 IIE
AREA tlK
3 . I ci
12 *-c-i-.3
3.5e3
9152 v
4. 115
4P,;?aei.; y
4 .*343
113196
v
---------
TOTAL
ic,28382
I DIIO
colic
2 . CII$83 1.4564 77.6958 18.81395 -------------
1013
O'Q
CD
Ln
Ln CD
CD CD 0
orq
CD
0 r+ (D
CD
(D CD
ID PC 0 tn CD CD < CD
START
I rj5
p 11t%i0
1
2 3 4
START
5 4.8445
4 11
T I IIE
AREA 111'% 1 DIIO
colic
3. 1 C12
16532
3.497
12395 V
4. 11
6362@Z" y
4.845
139 1 sa v
---------
TOTAL
8134309
2. C'554 1.541 79.0993
17.3843 -------------
p 11%110
1
a 3 4
START
4. 63e.
4. 105
T ItiE
3.498 4.1135 4.338
TOTAL
AREA 11K
11329
7764 V
441513
V
-1L42974 V
5 6 3 5 IL1I3
IDNO
colic
a. ci1 C13 1.3776 78.3407 18.2714 -------------
1013
P.1%,110 4
2.538
5.258 T IIlE
4.468 ARTA tiK
1 54E,6
3.307
13754 V
4. 4 6 *:.' 4 7 9 14 '-"* Y
5.258
1 1 19 1 cl y
---------
TOTAL
6213218
I DNO
colic
2. 4;339 2. ;2176 77.254-S 18.0437 -------------
1013
cn
cn C')
En
=r
CL
"3
m
0
C< I
r+
0
cn 0
r+
CD
CD
tn
x
lic
0 cn
go
a
C+ Cl>
CD
(D
<
CD
0
CD
>C
CD
START
P)'%N 0
2 3 4
START
4@- .7117
3.gee
T I ME
AREA
111'% 1 DN17i
colic
769
3.39 3.9.-'3
113513 '-1 42 7!f3,
4 .7ri?
136700
V
-- - - -- - - -
TOTAL
309fie 1
7 i.
1.4648 7 4 50 1 16.8974 - - - -- - - - -- - --
cn c@ CPO C)
cc 0 @-I a
lw aq
3.93 CD x 10 0
Pl'%NO
TIME
AREA MK IDNO
cc lit,
1
2.997
2
3.385
17 9 L,8 13222 v
2.3522 1.729
CD (D <
3
-3'.98
557714
Y
72.9303
4
-4.328
32223 14
4.2137
5
-4.693
143575
Y
---------
18.7747 -------------
START
TOTAL
764721
1.773
0 3E
Cl)
144
x
P1.11%113
T 111E
AREA
liK I DliO
colic
co
:3. 3.393
49 2rj365 Y
m
1
<
CD
3.1773
3
3.98,S
482711 SY
75.3127
4
-4.7
117317 v
---------
18.3039 -------------
T 1)T A L
64rJ942
lee