Document npapjrpmwpw037py5roZk0yo6
AR 286 _ 0778
BIODEGRADATION (301E)
IestsuesTANCE
0000
Identity: ArefmeirxrteudretocaonstPaiFnOiSng, pFeCr-f9l5u,oroorocatsanaescuolmfponoanteen,twhoficFhC-m2a0y6.al(s1o-be Octanesulfonic acid) (CAS# 2795-39-3).
Remarks: `sample is
The FC-206
3M production lot numberwas 2330, 10/79. (FM-3824). Current information indicates it
The is a
test mixture
of
0Su.l6t7o%nePfFoOaSm,er1,71.%5%sdoideituhmyloecnteylglsyucloflatbeu,t0yl.0et4h%ers,o7d8i.u9m1l%auwrayltesru,lf1at.e3,30%.5%
polyoxyethylene monooctylpheny ether, and 0.05% benzotriazole.
cThhaerafcotlelroiwziendg scounmcmenatrryataipopnlsieosftiompaumriitxiteusr.eDwaittha imnacyomnpoltetaeclcyurately reflect toxicity of the fluorochemical component of the test sample.
METHOD
0000
Method: Modified Testtype: Ready
OBiEodCeDgrSacdraebielntiyng
Test,
OECD
301E
with
DOC
Analysis
GLP: No
Year Completed: 1980
Analytical monitoring: Dissolved organic carbon (DOC)
Statistical methods: Results were removal and graphic interpretation.
determined
by
calculation
of
the
%
DOC
TTehsetsoercgoanndairsym esfofluurecnet:waAs5f0r:o5m0 amnixacotfivsaotledexstlraucdtgeanadersateicoonndbaasriyneafftltuehnet,St.
Paul, MN MetroWastewaterTreatment Plant, of White Bear Lake, Ramsey County, MN.
while
the
soil
was
from
the
city
Test condition:
Dilution water: Deionized water. Mineral Nutrient Medium: Nutrient medium per OECD 301E method
-
(1979). Initial pH 7.0.
Tpreesptaarnedd bryefdeirsseonlcveinsgol4u2t6iomngprFeCp-a2r0a6tiionno:nTehelitteerostfmmianteerriaall nwuatrsient
medium. This solution gives a final test concentration of 40 mg
DdiOsCsIoLlv.inTgh3e0.r6efmegreinncoenseublsittearnocfem,inheyrdarloqnuuitnrioennet,mweadsiupmr.epaThriesd. by
solution gives a final test concentration of 20 mg DOCIL.
Test vessels: 500 mL Erlenmeyer flasks containing 250 mL test
solution and capped with foam plugs.
Incubation conditions:
TLeigmhpteirnagt:uCroen:st2a3n.t5 d-a2rk4cConditions.
001415
Agitation: Continuously sNuubmsbtaenrceo)f acnodncbelnatnrka,taillonisn:dup1lipclautse.hydroquinone (reference Imneoacsuulruemmecnotnsdiatrieorneoqunirteedstpeinrimtieatthioonds:reNfoetreginvceend..)(No Element Basis: Decrease in dissolved organic carbon compared to
the blanks. Test substance flask conditions: Not given.
J
Nominal concentrations: and at ~20 mg DOCL plus
Blank HCl,
control, hydroquinone at ~20 mg DOC/L (inhibited, test material at ~ 40 mg DOCIL
aEnldemaetn~t4v0amlugesD:OCI2L8-plduasyHdgeCglrya,daaltlioinnd=up9li1c.a4t%e..duplicate 1 and 92.3%
duplicate 2. Mean value = 92%
RSuebmsatraknsc:e TReesmtianrgkwsafsielcd.onTduhcetveadlouenstrheepmoirxtteudraepapslydetosctrhiabtemdixitn utrhee aTnedstnot the fluorochemical proportion alone.
c0 onclusio0 ns 0000
The was
test substance % degradation 92% after 28 days.
based
on
the
mean
DOC
removal
value
Submitter: 3M Paul, Minnesota,
Company, 55133
Environmental
Laboratory,
P.O.
Box
33331,
St.
p0 ataQuaur0 y 000
Reliability: Kiimisch ranking 2. testing. However, the sample
This purity
study meets the was not properly
criteria for quality characterized and
the
study lacks in solution.
analytical
confirmation
of
the
amountof
fluorochemical
proportion
-
RE0 FEREN0 CES 000000
TLhabe sRteuqduieesstwenruembceornd5u5c4t1eSd, b1y98t0h.e 3M Company, EnvironmentaLlaboratory, ot0 wer0000000000
Last changed: 6/27/00
001416
f4r6an
TECHNICAL REPORT SUMMARY
TO: TECHNICAL COMMUNICATIONS CENTER -- 201.20N
norant = If seipproinrtedton both siodfpaepesr, sendtwocopies to TCC.)
FLCC) or
6/30/8
"iinvironmental Laboratory (EE & BC) Commercial Chemicals Division
0535 9970012600
+ Diodegradation of "LIGHT WATER" Products in OECD Test-6/80 | age
John A. Pignato - Commercial Chemicals Division - 236-24 (01)
Lric A. Reiner - Environmental Lab (EEWPC) - 21-BW (63) 47818
[NoP ne - r See 3M
Environmental 0 cous
Lab
Request No. 5541S EET
31 Err]
KEYWOSRDS:m a
CURRENT OBJECTIVE:
Smear
Environmental
TpTerosotduuscewtistthhearDeOiCnhtiaegnrhanllayytsibioisnoadltelogyrsahrdoeawcboltgehn.aitzed"LOIEGCHDT WSAcTrEeReSning
(LEubEor&atBoCr)y,
wr oc
SSeI, ASSTCAAoCTn: 3R2R02S50 wort Toi sore reat s Ge oyoeTd R Soo Se
SADSnnoLianIslcGylHsuTsistsiWuvAdTeyElaRynu"dsdeedBsmruoaptnnphsdletermA"aePMtnPoetFdailftiheepdaerxaOtlEelCneDsliSvcserteeebrniiiolndegegcrToaendsetaronliwsiitihtnyoDoOfC
leCv,3e0l1s7.of degraded.
FICFn-C2-1204063dadyessgh,rowaedtdehde9p3dbr%iyosdsuD9ocO0%tlCsv:edadengdroFaFGrd-ega2eat03ni30i,o1n7cSawiCnerosgs0o26n39.4s(soee))
mjnTaaohtsteeorrdpipuaaterlia.oltnlo,elchveosmltiaectraiillliezoarctoipnohtnyr,soilcsoarlpprprorevocecidepsitstheaasttiostnhaieosf oDoOthCe Ldpaaynaser.enwtas
~
BintEormRaonR GS
001417
)
-2-
BIODEGRADATION1O0FOE"CLDIGTHETSTWATER" PRODUCTS
LurionuCTION
ue 4M Enviconmental Laboratory chose to attempt to dedonstrate the biodegradapility of
method because the
"LIGHT test has
WATER" wide
products using the OECD international acceptance,
screening and the
Environmental Lab participant in an
has had experience international ring
using test.
the OECD test as a These results will
comp
high
lement
degree
the
of
beixoidsetgirngadaBbOiDlidtayt.a
that
show
these
products.
to
have
a
HETIODS AUD WATERIALS `The OECD
carbon.
test For
is run at 20-250C and it. [measures these water-miscible "LIGHT WATER"
not expected
only dissolved organic products, which are
pdteoegrsrttaiidonangt,iooftnh.ethOetTLosheCeDvpoTrltOoaeCdtsuitclaitnizsase.l,yasnipsarpecpmierptoihptoraditaete,evenmoermtehbaoesduardfesosorrbmteehdeasupdrueirrnifgniguo%ritnosteasld
afSftfcotfrlahelcoeohdnwesiidn"nggan(dTtAehpsCeptoennSddweiiipxttthieom1nb)D.esOrC-A1n9W,aaldye1s97iA9s.,(1S)cv.heerisliAonpcoeproyffoortfmheedtMhiotsdhiimfseicethdeosgOtiEnCsgDo
Ueductions in the scale of the test were necessary to adapt it to
acveapillaacebdle 2s-hlaiktienrg felaqsukisp,men2t5.0 wmFl iovfe hiunnodcruelda-tmeld nEurtlreinemnetyesrosfsleaisnkss
replaced
pfrielstcrraiteb
for DOC analysis. ed900promcl,eduarned, thtehe saanmaplleystsizuese
w d
as 25 only
,
not the
30 last
ml. 10
As in the ml of sample
A calivrated
within
chart
recorder
monitored
the
temperature
which
stayed
period near
the 20-259C range during when it reached 25.50C.
the The
28-day test, except for a 3-4 hr. temperature was most frequently
the high end of the range (23.5-249).
Chemicals - All chemicals were reagent grade unless otherwise noted.
OTfheAn3tMwe"rLpIGmHaTnuWfAaTcEtRu"rep.rodTuhcetisr ulsaebdelweriedenstaimfpileeds
of commercial material them as PC-203
FC=-3017 Lot 2303, and FC-206 (FM 3824) Lot 2330 10/79.
NFP,
`Ine hydroquinone used as yrade hydroquinone 98.5%,
the calibration compound was Lot 100147. Pour chemical
Aldrich reagent supply houses
apwevrrpoeasrcercaiotbnyetdacntooteyd trheieandiMayondiaafvtiateiedlmapbtOlEeCtDoionSbctrtaeheienniUnStgAh.eTe9sW9te.,5%exbpupetucrtethihtsyhadtrpouqriutiynonies
irewdpuucreidtiepsurihtavye wniollaphpaavreennto their presence could make
signficant inhibitory
effect on results.
this The
effecton microorganisms and
only a trivial change in TOC measurements.
001418
/
-3-
yga$ans0sioeouEnSgti.ncuserPdaeusliinn,cibttehdye, wpaartnedeprar2paamtsiisxoeenddofbtehdraolludgehisoonaliuztceiarorsnb,so,nabneedxdc,etphmteanctthfeeiphoeirengoounceyulum,
eCoxpecrvaipmoern2,at0t;ieon2ns-.uurnedSfticolotcenkrss.isotfeTnhwtiastwerawtaetcreorlqluewacalstietdayl.saot utsheedsttaortmakoef uspheLosses
t4C5hex17'i:0r5p5e,Pee3Ebc.oaaCacLaciCEnlygb.ednuht3r3e.is4eengtroesfpolluaNtciielomPneOsn4,-t2si21y63i.0e4,ldge3dO6f.3soal4nuhtOyfidornoCasucslayaN:daAyliopi0ipnOgcerroeepnipeelnaaccciyeodn
Se$oxo4t1Ur85aiscbotan2s(co)olt)nuo.stiitsohtnToehsedewas1r0e0quusmeigdredoifnbPypel-atcchheeelOoafEtCeDthueMestevhdiotdai.nmintThehSeoltourpapctieieoyneslsemeynetast
and 70 ag EDTA). of equimolar amounts of FeCly and EDTA (30 mg FeCl
ZSeesrtveCdonacsentthreatbiaosniss approximately 40 mg
o-ffoDrOcCaardbjmuoesnatsipunergremelininttiestri.aolf-Tsthaeimssptleciosnscttheoenctkrhaidtgiihloeunststionrsg
Shmieoganhscuernietniiretanittaislo.ncoiTnnhceetnhetlroawrtaitnoognxeicriinetccyroemoamfseendd"eLdIthGeHbTysWetAnhTesEiRtO"iEvCiDptryotdeoustfc.ts
UDsOeC so
ovfemetnheiys
auincliekoeolryy.anisns made significant inhibition at this concentration
2ccao3rnJbcpoesnnttarepanettrioolnfithoefyrdrw6oa5q.s4u%id.noonnee,bythweeicgahltibbraasteidononcoimtpeouKnndo,wntoca2n0onmg of ucoerrebcltainokncs.ontrols, instead of 1, were utilized in making blank DOC
1SSTsfo:aig8gnS.dey2grl.'u1snagnadrTdh1ee.n6.ia2nn.od2cuotlfhuemtwshauespeOrE5nC0aD-t50atnetmsitoxftusuiranegs'apmrfperlpeeasrherdsoomrascacnoerrdaaicntsgi,vatto3.edsteps
cuhsledoriwnasea-efWrrhaeatetiwoanwneRlblas54wi,anteaarnt.d tthhee Swt.atePraulforMetsrooil Pelaxnttr.actiTohne
filter
was
paper
S3sieclecCooSterilennttrorlosl-sRefpolriceaatceh hSaonmdploiunnd. Handling Of these
svtteeessrstiellmesateccrooinnattlraoilnasinndgwtahs4e00icdmaegnl/tilibcraafligiCsliyo
de cved the
gU-anealndingg28o-ffdraeyqtuheesnacmvypi,laebslb.uetcutlhteuraensa.lystTheonylyhamdadteheDOsCammeeaisnunroecmuelnutnsamayu the
fSgSadromuebppsllceeorfivPecrdoenscbeeyrnvatttrhiaeotneTdOC- HTCmnLaenutfosaacmtthpuelree1r0p(r4me)ls.erfviTalhtteeiroenmdemteshtaohmdpoldiensuv,soeldbvreiswnagsaidndtgihnagtthe1 wpSihrtaehpsielraivlzautetmoiion<pnulmlmef2t,ohioladn-.dlinseTtdhoericnmapgasn.utfhaecTthseuarmmeperltehsiondduincdiseartaensreeftfhrfaietgcetrtihaveteimoenthiondvciamnis phehalate, caalirperaadtiilyonbisoodluetgiroandsablmeadematferroimalp,otafsosriusmevehryadlrogmoennths.
001419
nSiencceessatrhye JSeesaecciiibcead)
asantcdeipdailfilniocwatehtedioTneOlCiimnainntaahltiyissoinpsroefpsreorttvhoeactoUilog,nlymietstphrouedsseerwwvaaasstiaoltnsheo tmaeocshtnique
month of thien ptrheesOeErCvDatimoenthoodf. thAeLLsamTpOlCesa.nalysas were made within
SsoonsrtmraunmenDCt-s32aAn.7d per liter.
cqIutispmseenntsi-tiTvhietyorLgiamniitc icsabrebtowneeannal1yzaenrd
2usemdg
owfascaarbon
apaSlniiuaenglesysS.stioasnp.dpGeerlssmtaonrfeodr0.2itnheudmermieeoanmcibtzrieaodnnewfalftaieslrkt,esrsws,eerrve(ePdcalrettaonH,of.ilpto6re4or8u1s4s,)ampbploleiaslsetdifcou3 fDoOaCm
C(tCahsesownaerryeeRc)leafnoilnlgoweidnvboylve6ddesiooankiinzgedinwactherromriicnseasc.id cleaning solution
M HESUELTSSAANNU DD DDIL ISSCCUT USSSSISON
cfaobnlteain| tshuemmaarcitzueasl ccoamnpclieotnelyo.f time.
AdtLahLtear3sehs"euLelIttGssHToafWnAdTtEphRil"sotsAstPuoPdFfy.ptrhoedAupdcpetegsnrdaiddceaegtsriaodn1esaasnndeaa2rly
Percent DegradPCa-t2i0o3n, ofFC-f2i0y6dr,oqaunTidAnBoLFnECe-31a0n1d7 "WLiItGhHTTiWmAeTERi"n AFFF Products
Modified OECD Screening Test
$ Degradation at Day:
product 7 14 2 21
1)
FCC--220036 1233 8905
993
9931
re-3017 37
94
96
96
9932 95
uqyudcion-one 89 92
97
9
93
.
gJncotryhbreeucncilapisiketealtoyfiotnhtahtceauhpsihegydhsliyctahelwatmleeorassnssooflsuubscloheluabs"lLeIadGTsHOoTCrpWtA(iTDoEOnRC,"). vproTolhdauitcsitlsii,zsatiitoni.s FgFCue-sb2tss0t6aOnftBiOaDtt5hee/dCO3DMby=Entv0h.ei5r)or.nemleanttiavlelyLahbiogrhatoorxyyge(nFCd-e2m03andBOoDbss/eCrOvDed=. 0i.n5,Bo-D SoSYvLliIoaGeeHnTrceWerAiTltEehRa"tcopntrthoreodluscotdlsautbalies imTnoOtTCadbluloeess t2ofprarodomsvoitrdhpeetiftouenrs,tthvesoralmasptulibelssitzacanottniitoaanli,ninoge sparmepcliepsitaetxicoenp.t tThhaetsethesyampcloenstaiwneerde hHaCnld,ledtoipdreenvteinctalmliycrtoobitahle gtreoswtth.
001420
.
aftLohet chaienmiateijnadolrofDfOaCct.thoer,T2h8ua-stdaylloesatssetsotffposreorilnouodbn,lmeettaThbOeoCylibszyteidltlhe"scLeoInGtpHahTyisWniAecTdaElRn"emaordleys awllas cunoeotmtpaobrnoeelmniotsvsme. o"cLcTIuhGriHrsTedWcAoTniEtnRr"otlhemsedetoaebssotnle'irttielpserovcieonntttrhhoealtst,pehsbtyustircuantlsh,eprcsoaictneacbseoslenisgctdid cfaotramvaotliiocn opfrodluecstsswaarteerussuoallubllyemomraeterpioallasr ainsdusnmlailkleelry. because
TABLE 2
DOC Loss from Sterile Controls
Product
%atDOCDayRem28aining
PCC--220036 C-3017
9914..32
liydroguinone
8787..15
-
Adsnalyiogfh0ft-lpywrhiiobtrreowtnpoirsefhci,ilptiaetsraitwnaegs ffotorhremeDdOsColiunatniatolhynesihisny.dtrhoeqTuhiehnyodpnrreoeqcuisiptnieotrnaietleesctwoeanrsitlreol on
cSOoiLncCtbDreoaelsxtpearfiintmeertnhte2.1prdaTeyhssie.snceapTpohfiasreNnsCtulgygt,eesntdtsehnrcotyuhgahtofouhthyyddrtrhoeoqquuicinonuoornnseee iOstfonotthest2a8b-ldea,y
sipnoanptparnoperoiuastley precipitation
wcfiootnrhtmrolilgnCslco,loumbpaloleusnodhcuamfsuotsrs-ltishkoemeeOmEdaCotDuerbtiteasslt.ambaokuIettss thipetartainal
ameptphroodp.riateness of using the igCly preservation procedure in the OECD
coucLusron Tpihnreod2u1pcrtdesas,yesntFCu-ns2dt0eu3rd,ytchPeoCn-2cc0lo6un,sdiivtaienoldnysFdCoe-f3m0o1nt7sh,etramatoredesifnieteahdratlOyE"CcLDoIGmSHpcTlreeWteAenTliEynRg"degTersatd.ed REFERENCES
(1) "o1Tr9e7gs9at,niinzTgeastPtirooGnguriafdmoerlEiExncpeoenrotfmoirGcrotCuheop-oMonopderiDafetiyierodandOaEtainCdoDn/SDAcecrvceeuelmnouiplnmagetnitToensC,thewmDiietcch.a',lsDOC Analysis (Level I) (H. G. Hosler) Revision of Sept. 19, 1979.
(2) ponrman DC-52a Operating Manual, 4th Ed., 1978, p. 3-1.
oo
Appendices: 21 -- ODEatCaD TSehesettsGuidelines (Sept. 19, 1979)
3 - Graphs
* mCoalteacbuolleiss.m is biologically facilitated breakdown to less complex
001421
Me
APPENDIX 1
!
Tokyo, Decemoer isc, 197g
C 118/79/Int.
OECD - Chemicals Testing Programme
Expert Group C, Degradation/Accumulation
Modified OECD Screening Test with DOC Analysis
001422
Test Guideline C 118/79/Ins.
Test with
GDuOCideAlnianlyesifsor
the
Modifid
OECD
Screening
Test
Date of last revision: Sept. 19,1979
Levl I test for ready biodegradability
1. Prerequisites sItoluhbalse toin bethkenocwonncwehnettrhaetriontherantgoest emmaptleoryieadl inis the test (corresponding to 5 - 40 mg DOC/1).
2. Guidance Information Kpnroowpleerdtgiecsofofthtehebatcetsetrimaaltertioaxliciitsynootr uinnehqiubiivtoocraylly creoqnudiurcetdionbutofcotnhestittesutt.es useful information for tho
3. Qualifyine Statements Tuhletimnaettehodbioisdesguriadtaebdilfiotrythoef wmeaatseurresmoelunbtleo,f ntohen-avoelraotbiilce oervgaalnuiactiocnompofounmdisx.turIets.is unsuited for the biodegradability
001423
Sse Stores Sept, 19, 1979
Modified OECD Screening Test with DOC Analysis Preamble -
It has to be realized that the following procedure for the modified OECD Screening Test has to be regarded in some points as provisional since several important features such as the calibration compound and the new test duration of 28 days are
completely untried yet. However, since the test
constitutes a modification the official, widely practised and well accepted OECD Screening Test for the biodegradability evaluation of surfactants there should exist a good prospect for success.
001424
Septemoer 19., 1979
OECD Chemicals Testing Programme Test Guideline for the Modified OECD Screening Test with DOC Analysis
O. The test procedure constitutes a modification of the OECD Screening Test (OECD Environment Directorate, Proposed Method for the Determination of the Biodegradability of Surfactants Used in Synthetic Detergents, Paris 1976, and council directive of Nov. 22, 1973, on the approximation of the laws of the member states relating to methods of testing the biodegradability of anionic surfactagts (73/M0S/EEC), Official Journal of the European Communities No. 4 347/53 of Dec. 17, 1973) for the application of the dissolved organic carbon (DOC analysis.
1. Methed
1.1 Introduction: Purpose, scope, relevance, and aoplicaLe tiioonx ooffp tteessl taana ndd en xpla anatt ioni ooffo lliimmn itss.
The purpose of the method
ultimate biodegradability
is
of
the measurement of
water soluble, non
the
-
volatile organic compounds in an aerobic, aqueous
mteodi5um- a4t0magsDtOaCr/t1ing( tIenstordceorncetontravaotiidonicnohribrietsoproynding
effects it is in the investigator's own interest
-2-
001425
-2-
to choose as low a starting concentration as his
analytical capability permits ) :
1.2 Definitions and units 1.2.1 Definition of biodegradability
of. || 1.
L
where
T Ge b - omeg |, gg,
Cc,o = C,bl,1
Dy = degradation in.percent DOC-removal
at time t
, = starting DOC concentration of the culture medium (mg DOC/1)
Ce
=
DOC concentration of
at tine t (mg DOC/1)
the
culture
medium
Cy) starting DOC concentration of the
blank (mg DOC/1)
.
Cpy= DOC concentration of the blank at time t (mg DOC/1)
1.2.2 Units
The degradation is stated as the percentage
.
DOC-removal within 28 days with respect to
the test material ( % DOC-removal )
"3.
002426
@
5
1.3 Reference compounds
1.3.1
Calibration compound
The calibration compound used in this test is
hydroquinone at a concentration corresponding
to 20 mg DOC/1. Hydroquinone has to exhibit a
DOC-removal of 260 % within 28 days, otherwise the test is regarded as invalid. -
1.4 The principle of the method
A predetermined amount of the compound is dissoi-
ved in an inorganic medium (mineral nutrient solution, fortified with a trace element and essential
vitamin solution), providing a concentration corres-
ponding to 5 - 40 mg DOC/1. The solution is inocu-
lated with a small number of microorganisms from a
mixed population and aerated at 293 - 298 (20 - 25)
in the dark or at least in diffuse light only. The degradation is followed by DOC analysis over a 28
day period. The procedure is checked by means of a
standard (hydroquinone).
A control with inoculation but without either test material or standard is run parallel for the determination of DOC blanks.
1.5 Quality criteria
1.5.1 Reproducibility
:
The reproducibility of the method is appropriate
-----
for a screening test which has solely an accep-
tance but no rejective function.
.
001427
-
1.5.2 Sensitivity The sensitivity of the method is largely determined by the sensitivity limit of the organic carbon analysis which is 0.5 mg C/1 at the present state of the art.
1.5.3 Specificity, applicability Applicable for the biodegradability evaluation of water soluble, non - volatile organic compounds.
1.5.4 Possibilisy of standardization The test version with specific analyses for anionic and nonionic surfactants is standardized as "OECD Screening Test".
1.5.5 Possibility of automation Parts of `the test, e.g., the analysis,can be automated, although hardly the total procedure. The procedure is, though, well suited for being operated with whole series of test materials.
1.5.6 Costs ( in 1978 Swiss Francs )
1.5.6.1 Equipment
Glassware
|
shaking machine
2000,6000 = 20000,-
|
Carbon analyzer
42000,
Miscellaneous (pH meter, balance,
provision for air donditioning)
5000,
-5-
I
001428
-5-
1.5.6.2 Person - hours
10
2.5.6.3 Approximate total costpertest 750,= = 2000,-
(aimnovretsitzmeedn)ts assumed to be
1.6 Description of the method
1.6.1 Reagents and materials
-
1.6.1.1 Deionized water
Deionized or distilled water `ree of toxic
substances (copper in particular), for general use a3 a solvent. Water which has been deionized by distillation or ion exchange is suitable.
A high purity of this test water is necessary
in view of the DOC analyses in the concentra-
tion range of 0 - 40 mg/l. The contaminations
result from inherent impurities but also from
the ion exchange resins and microbial devel-
opments (bacteria, algae under the influence
i
of light etc). Only one water charge must be used for one test series which is to be controlled
1
b
beforehand by DOC analysis. If necessary,
suitable water may be gained by UV irradiation
-
{{
or other means.
{
1.6.1.2 Nutrient solution
|
Mix 1 ml each of the following solutions (a) to (f) and make up to a volume of 1 1 with
water 1.6.1.1
-6-
6014293
-6-
(a) AHLPO,
AR.
8.58
KHPO,
AJR.
21.758
Na HPO, . 2 H,0 A.R. 33.4 g
NH CL
AR.
20.0 8
~
in 1000 ml of water 1.6.1.1 the pi value should be 7.2
(2) 22.5 g of MgSO, . 7 HO A.R. dissolved in 1000 ml of water (3.3.1)
(e) 27.5 g of CaCl, A.R. dissolved in 1000 ml of water (3.3.1)
(4) 0.25 g of FeCly . 6H,0 A.R. dissolved in 2000 ml
of water (3.3.1) This solution is prepared freshly immediately before use, (e) Trace element solution
Mas0, . 4 H0 39.9 mg (30.23 mgMnSO, . H0)
Hy30,
57,2 mg
2080, . 7 H,0 42.8 mg
(NH) gMo,0,, 34.7 mg (36.85 mg (NH,) gl070,, .4Ei 0)
Fe - chelate (FeCly,EDTA) water 1.6.1.1
100 ag 1000 m1
Sverilisation of the trace element stock solution
as 393 (X) (220C), 2 atm., 20 min.
-7-
0014350
@
-7-
() Vitamin solution
Biotin
Nicotinic acid
Thiamine
p-Aminobenzoic
acid
0.2 mg
2.0 mg
1.0 mg
1.0 mg
Pantothenic acid Pyridoxamine Cyanocobalamine
Polic acid
1.0 mg
=
5.0 mg
2.0 mg
5.0 mg
water 1.6.1.1
100 ml
The solution is filtered sterile (0.2 pm). Instead
of solution 1.6.1.2 (f) 15 mg of yeast extraxt
may be used per 100 ml of water 1.6.1.1.
1.6.1.3 2Hyidordoeqguriandoanbeili9t9y,5s%taDnAdBardErg. B. 6
1.6.1.4 Mercuric chloride solution 1 per cent of Hgll, in water
1.6.1.5 Shaking machine accomodating 2 ltr, Erlen-
meyer flasks either with automatic tempera-
ture control or used in a conrtant temperature
room at 293 - 298(K) (20 - 25C)
1.6.1.6
Narrow neck = 2 ltr. Erlenmeyer flasks. (Creased
flasks are recommended) The flasks must be care-
fully cleaned with, e.g., alcoholic hydrochloric
fluted
-8-
001431
Sa-
po
~@ use, rinsed and dried in order to avoid
1
wnation with residues from previous tests.
+ .lasks also have to be cleaned before their
.FSt use since they may be contaminated.
1.6.1.7 Membranefiltrationapparatus
1.6.1.8 Membrane filters 0,2 um
1.6.1.9 Carbon analyzer
1.6.2
Incculation Either of the following three alternatives may be used 2s inoculum or a composite sample thereof.
1.6.2.1 Ino:ulum from secondary effluent
The inoculum is gained preferentially from a
secondary effluent of 8ood quality collcted from
2 treatment plant dealing with apredominantly
dorestic sewage. The effluent must be kept under
.
aerobic conditions in the period between sampling
aid use. To prepare the inoculum the sample is
filtered through a coarse filter, the first 200 m1
deing ciscarded. The filtrate is kept aerobic until
-
ised. The inoculum must be used on the day of collec~
tion.
1.6.2.2
Inoculum from soil 100 g of soil (fertile, not sterile) are suspended in 1000 ml of chlorine-free drinking water (soils #ith an extremely large content of clay, sand or organic carbon are unsuited). After stirring the suspensicn is allowed to settle for 30 minutes.
to
001432
-9-
The supernatant is filtered through a coarse
filter paper, the first 200 ml being discarded.
The
The
filtrate
inoculum
miusstaerbeateudsedimomnedtihaetedlayyanodf
until use.
collection.
1.6.2.3
1.6.2.4
AInnociunloucmulufmromis adrsauwrnfacferomwaatersuitable surface water.
The sample is filtered through a coarse paper,
the first 200 ml being discarded. The filtrate is
kept aerobic until used. The inoculum must be used
on the day of collection.
Composite inoculum
Equal
mixed
wveolllu,mesardofthtehefi3nailnociunloucmulusmamdprleaswnarferomunittheids,
mixture.
tT
The suitability of the inoculum is checked by means of the standardhydroquinone.
1.6.3 Conduction of the test 1.6.3.1 Procedure
The test materials are evaluated simultaneously
duplicates together with the standard (1.6.1.3)
in
a control test with inoculation but without
ana
test
or
standard
material
for
the
either
determination of
|
|
DOC blanks.
The standard material has to attain 60 DOC re- -
i
mcoovrarleswpiotnhdiinng 28toda2y0smgatDOaC/s1t.arItfin(g60con%ceDnOCtraretmioovnal
|
are achieved the whole series has to be discarded +)
||
+) Tthhies1T9IRdIaTy ivserbsaisoendoofntphreesteenstt.eXAperreIvSisRiCoEnWIofT --
||
|
ttohisbe licmointsidoerreedveanftoefr thtehe satcacnudmaurldatmiioghnt ofhaevexperien-
ce with the new 28 day version of the test.
- 10 -
005433
"10 -
A(1.s6t.o1c.k1)soilsutpiroenpaorfedt.heSotemsutchmaotferitahlis isntwoackter
solution is added to the nutrient solution
m(81.D6O.C1./21) isthaatttaainceadr.boTnhecosntcaernttirnagtiocnoncoefnt5ra-t4i0on
of the
poc/1.
standard
hydroquinone
is,
though,
20 mg
Twiwtohre9a0c0tmilonofvesthseelsnut(r1i.e6n.t1.5s)olaurteioneaacnhdfiilnloecdula
ted
open
iwnigthof0,5theml/v1essoefl
the
is
cionvoecrueldumwi(t1h.,6.2e)..g.,Thaelumi-
num foil in such a way that the exchange of air
between the flask and the surrounding atmosphere
is not unduly impeded (Cotton
because of the DOC analysis).
wool is unsuited
The vessels are
then inserted in the shaking machine. The tempe-
rature of 293 -298K ( 20 -25%) must be maintained
unchanged during the test, and the vessels should
pbeollsuhtiaenltdsedanfdromtoxliicghtm.ateTrhiealasir (schholuolrdinabteedfreseol-of
vents ete).
cInonctehnetrcaotuirosnes oafrethedetbeiromdiengerdadaintidounpltiecsattethe(1b.o6c.4.2) at the beginning (day 0), and on the 27. and 28,
day. Three additional analyses have to be performed
-11-
001434
11.
in rather ~21. day)
regular
time
intervals
(~7.,
~14., and
The analyses are registered in the attached form sheet and evaluated.
Only the necessary volumes of drawn for each determination;
culture medium may however, they have
be to
be large
fugation
enough for
preceeding
the
the
cmaermbbornandeetfeirlmtirnaattiioonn.orThceentlrait-ter
requires differing
Evaporation losses up by adding water
vooflutmheescuflotrurtehemeddiifufmereanrte (1.6.1.1)
instruments.
to be made
in the required amounts,
The culture medium is to be mixed well before withdrawing
a sample. Material adhering to the wall of the vessel has
to be dissolved or suspended before sampling. The membrane
Tfhieltrfaitltieornedor
orcecnetnrtirfiufguagteidonsahmapsletso.
be_doneimmediately.
have to be analyzed
on the same day, otherwise they must be preserved with
0,05 ml of
of nutrient
the Hell,
medium or
solution (1.6.1.4)
by storing them at
for
2
~
each
4.
10ml
The
bxihoidbeigtrsadaabdielgirtaydatteisotn irsatvealwiidthpirnovitdheedsptehceifsiteadndraarndge.
-
Tmhienertaelstizactainonbe
finished before
is accomplished.
the
28.
day
if
complete
All steps require
Pipettes etc. but
great care and
not sterility.
cleanliness
of
the
vessels,
Te
001435
"12 -
1.6.3.2 Calculation of results
The degradation at the time t is calculated
.
from the determinations of the DOC concentra-
tions at the beginning (C,) and the time t (C,) according to
Doz
T
| 1=
--Ccm - Co0 r,Et | [x10
i
"Car |
where D, = degradation in per cent at time tT C, = measured starting DOC concentration of
the inoculated culture medium (mg DOC/1)
C, = DOC concentration of the culture medium
at time t (mg DOC/1)
Coa, = starting DOC blank of the mineral nutrient
solution with inoculation but without test
material (mg DOC/1)
:
Cyy, = DOC blank of the mineral nutrient solution with inoculation but without test material
at the time t (mg DOC/1)
The degradation rates are calculated to the nearest
0,1 %. The means of the D, values are calculated and
|
reported to the nearest full per cent. Results
"13 -
001436
W.
13 -
ending in 0,5 are rounded
number. The course of the
up to the nearest whole
degradation test is
followed graphically in a diagram as shown in the
attached example. The results are reported on the
attached data sheet.
The results of the degradation test are valid condition is met that in the same test series standard yields = 60 % DOC-removal.
if the the
1.6.4 Analytical means
1.6.4.1 Membrane filter 0:2 um, 25 mm (1.6.1.6). Pre-
paration of the filters: membrane filters are
impregnatd with surfactants for hydrophilization.
Thus each filter contains up to several mg of
soluble carbon which would
radability determinations.
interfere
Therefore
in the biode-
the filters
are purified from surfactants and other soluble
organic interferences by boiling them 3 times 1 hr
each in deionized water. These filters may be stored
in water (1.6.1.1) for at least one week.
Other membrane filters are suitable if it is assured
that they neither release carbon nor adsorb the
compourd in the filtration step.
-
If the samples are centrifuged, this has to be done
at 40 000 m secTM ( 4000 g) for 15 minutes, preferably
in a refrigerated centrifuge, in any case (40C.
(Remark: the differentiation TOC:DOC by centrifuga-
tion
work
at very low concentrations does not seem to
well since either not all bacteria are removed
-
or carbon as part of the bacterial plasma is redissolved.
At higher test concentrations ( 310 mg C/1) ard the same
small inoculation the centrifugation error seems to be
comparatively small ).
001437
ZF0oraSShheeesttfoorr tthhee Meomdriffaaeudd O0C2DDScSrcreeeenniinnggTTeesstt
e
a p enrou:start of was TTehsctors/tisceaalncatreastmactaenrei.:al: Gtanrovcouniuna:nalyzer:
_-- eee E-- E -- F-- S------
A: Concrols
1300 8351"23d1 1Tuc0 ion enc sorucion
Tc" 1as pacTM*
u: Carhon docersinacions: Culture seaiua
her mr
ost ators
TeTy"ses co=nTfea.e fo copyR En7eTnhiE jam wm " ja
p7
[I
a -f2getsToTTTT
Cer
fPaoglorinstseemOucnrisotnc
[aa
9 2
[TT
a= SavSe | /
Cop)
: Evaluation of rov ssa:
iloo - cancantrataons
= nor
heme
2 fp, -22=f3
|[ [O] TBoEM Feel atm er mGT a i
sean 0-, 2+02, .
i:
"CSRs heenae PO and TOC wadtoirn ananybnxra Soars NAETIONSSE 30ELSES OF The tearS044
wr
.
-1 -
1.6.4.2 The DOC measurement
The sample withdrawn from the culture medium
(about 30 ml) is centrifuged or membrane filtered
immediately in the
using the membrane
filtration apparatus (1.6.1.6)
filters prepared acc. to 1.6.4.1.
The first 20 ml of the filtrate are discarded:
.
The DOC concentration is determined twice in the
remaining filtrate TOC/DOC instrument
(about 10 ml) (1.6.1.8). If
by means of the the filtrate
cannot be analyzed on the same day it has to be
conserved ace. to 1.6.3.1. The DOC (mgC/1) obtained are registered on
measurements the `attached
data sheet and the DOC concentrations of the
culture medium and of the blanks calculated for
each sampling time.
001439
PE APPENDIX 2
-
rent31sEnovFirtohnamBaantda1)60Labo0r6a00toreyravtatianghaTsotse
:
ALSonabtaolR'yeotqf:uesSattadrNet ooS:iA eFenSr5o41rT S ES
BEST COPY AVAILABLE
eC(IohnrenbocarrenetiM:ocTayllcso9o0rAl6a]rSDoaIorvndreNtrantosin<WheBairoErrieArslntVeEorrBaPeaNfsEtapPRerLntEUCSTaFrAlNbTueECntas,eafnhdemeerrd lmactelroismolcapalllswes_btyotursotseuirnons, 38.5%, Lot 10007, Atgeten
vestenacer Scostmnt olone
A. botecatnation of vic! of Yost meterion;
Athstocdkatonsiozleudtiwonatoefrthe tase sstarial was prepared by diluting 117O mp to a final vatimw of 500 a1
SIn ShTock smolutTion.| stock solueion| cast m=aterial| Tbhiinstreasfulttowneesmsuresneisds to caCalcuatle culatethtehenoecceessssary
0. Carbon oaterinations:?
-
Winerat mureient]
GHonde.lof tase|Cfovcautacred 0a0 0D0): con7e. stirA days dToecunbeitise, 237]en, FaJZzlFarlTesrzzTae tr
ogrs ige
i| wo S E EEEREr EIEE
Cm Ther [oy [ore |22[roo E PEeEFEEEECEGD Fe far]
TE lear fy EE MSineuralpnsuturlseent]
roc
|e oeteem TFe]e aoosxr [F1L2 57
Treo
[oso
1 E FyErrSre eto riers
Ca eee [Rr
--
Coie = [1% 00 [0d 112 T06] <
Toon Joi [ne[arn] me--|SEEP [Pict|n eye o i|ge psi sr
[=[a Jes os ai[as] ear vere vied na carcutetions
[OE RIRAL)
1. Tha 2.006
atneasltysmeastaprelraflor1msedcommoprleettehleyn
stouliucbeleonInthweateserm.e
sTahmeprleefowraer,e
trhuenTIfEor =qu0a6l,ity
assurance
purposes.
001440
BETA BEST COPY AVAILABLE
p eere e erat Lob Request No.:_ S541S rorstn m/z Standard Materials hydrogioons, 30.5%, Lot 100W7, larien
RPerteeToeBl aTorrt earire EGrEBDLCT
c
A. Qotermination of Toc! of Test Material:
A stocokmmseonludtiownatoefr the test material C pamr soutiT en [sock0Sgucon|
was prepared
sonvimeonrtal
by
|
diluting L4o06t0e mg to & fine] volume of EBs S00 al aTo OLnLtS3OSan Ant a1b0is0s4t5 SSoHpIaVhiepCohaiers
0. Carbon Determinations;
-
Ta | TEore a sn|| T SEEW ri Re aT,
shtnroovreions
nis
aded Maorel nutrient]
Inoculum
gooleofottevipooten:
412
Ee Yo.0 EEE
Ee ee
2
Fett
TE elle eer 426
Yoo
EErE eee or e E[ B ERA Ay
TTT
rTE are eae 1 El Err eer ete Saoee eTto erv <
T= RLTa er] FA eee EEEEE ET TeTey =
er LeeTe28
Te[e we[f5a |a77n temoro Te ae caerds
[= [23 [Fo[9[95] 92 | Va* 2
21.. 0T0hCe acnoasltysmeasteproiraflor1nsedcommaprlsetethlaynsteoilceeGnfnthweatesram,s TSamhpleawrareathFfuenTof0orr=sq0u0aK,l,ity surance purposes. 001441
wrenchesarthe Restf16d kisSreseiosTost BEST COPY AVAILABLE
Bs a, a SreeScandard DLoabteRoefqueSsttartNoo.:fTe5s5t4:1_SS-1[-80 ~ tatrtat:_r byironso oene, s 0.90.5%,08 Ls oLott 1000o 017M,, Bpir, eeriiccnn a TorS aeRe e ered Be RASL nTtERR me a) weston cenit stan
A. otermination ofToc! of Test Material; A stock solution of the test material wes prepared by diluting L2YS ug to a final volume of S00 ol
R ThohiT n [scot1seueion| neA vegerer | BTs h fresh wn aus luitd c2ociaE cleetcuheetccoesstsasry
0. Carbon Determinstions;?
F| T Ee Ea TR TEE Wer? ruteiont
solution with
frogs
-
TTT CT ann [3et
So TLi
20%
Yoo
mes
2033
20., 0
| fewfealar[sez| Ete IEYIpR2E8PETeT30 EEET a 25 J a 5c
I
----
Cer
esep Re
eeeu eo]
mE a mass [enfae 0 a ten oral nutrient]
EE = ale Tae ha
i [[T TouTuo aTor]- C. Evalustion of Maw vata;
em Ce e oo] r erro o9 r1
HerFt aee eer e =
10
Cae [19 [-or oy [72 [oe
[Tor oCpee rpiTeEtFRO iSFotnn ST XOGo TRI] | gsue cacmpd t ss ection t 1608 s for
| [oonToselosr[ona]
[=[za [aaa[asaoes]
sea
ave
|
|
oT p m L-am euas=ass ooecasteminiones
[~~17ToTae[ae | ar | lave--p-- i
2.1. 50T0hkeaantnaeatslytseatsepraiaprolrtfe tos crmomanprlseetdtelhayntssoeullucbaloenftnhveatsara,masTphaatlreefvoresr,ertuhnefToer=Se0l6F.cy assurance purpoo ses. t
In Environmental Loboratory Data Sheet
Co tenn ows 35b1
BEST COPY AVAILABLE
MA reateraietse)hTtouseRO3.TSoYneeE reBrn R ERe AACeTATTGT A, Aunr T " R r--
Te] Eg | TRH Cnn A. ElSe Qoterminstion of Toc! of Test ons Meterial; sae bp HI een 4 1YioF i
TEETER TEE) icente0w sti ccna
TT evmutrion or caustans . Coron Determinations?;
aEET rh a wSam Minors mereiont
CommCo T E Tis E mepop E asrJluvseJo sn rie)
COMPOUN--D SEE STANDARD
MATERIAL ABOVE .
Sha [ot
30.6 | 20.0 lF Epoe HEE 1 p 70 1e r3p 7i1 0er3g7 i ae1r7 3 i3e 17
| 20.0
E E t Te es r ee
Sr geTye1
1 ----
1 epHerter
rests
C.. EEvvaelluuasttiioonooffRRaarwO0s:ta;
TE TST [oT v[ra foc]- Fa AerTee er CCrave == Cu, L+ oCum,vs 9 19 Joool)0d | 1.2 [eb
[Pa oepiiona3 ontoie OgTo| sagscien13tr
|[ses[res[soe[m7] gro | tr msoh tdimsmReoazmotsn
[==Te [22 [97[ov[93] se 01
A The vase sevoriat 1s cmpivci ay SolitaPIn wp are Tartore, onnN7s0 h06,SE
001443
mEiisEEEs E F E E eee e Ea e rs R ETERS
gy tee woe pets e Ae InalysGt:e VoNde A.I e Sche|l fetes wee
sh E Ea m a Sooo -
E = ee 0
=
-- =
=x= | ie oZf
i=== et
eee
eetr --------"4 |
EEE
Ee
EE Em fo So
.
aire hes
Cm
hme=
= Lash
EEL E ey -- o --
cen wen Malan 020Modifled ScreeningTestwith DOC Analysls
: " Se=em
= SII ITT
Date of Start of Test: 4e11-1980
E+R EEiTEo R
eUe TES
un
weiAreseeams
fessepastes wm'
oo SE
Mee
_----
ee
= --------i|
he
= --=
Eee e e
nee
eee
a P =e EeT e eeee e F e y eE e] e
24
avs ow
--
EEE E TEm Ee m Ee
ect mama
ubeA
Redan es
a Ega p E
a -- eee-- eet
mi EE=----Pi pre------
Ee
= 0=
: See
30
= & mmr
mei Soa ms
III LIIT 1
mn
i.201 =T e = e e ee m e e ere ] s ee p a e eey e
SH
eee seemnba py
s
Ti es nas 34 Environ-ental Laboratory.
oe
Emenee o Eme mbee cea EINE Aenaslystt:aWande Ao.f STcahetllbette
EEm EEEzA
"Ee=------- er a
Test Material:MYORoguONE
cnupoation_compounin)
E Titei nGa n
w= ie a ete
= =m
eae bre
E E ee E sass
oy
----
----
--
saree baTo MS
--------------
=e ee
ET ee
eee
Q BP R H Er f E e nee e rea e a e ae
~N
NUKSER OF DAYS INCUBATED
ul L
D Sec | @ @ HFoEx REPwSo. woox- aon
feYer! pe - 206206 0
PRODUCT
REQUEST
ECOLOGICAL ASSESSMENT
FOR LABORATORY WORK
ts ePianR traenO scstipenB fsmsav.ocU, =SBIo DL6o]FC-e293
Las reauest3io._ S341 S
X 40
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PE aaFT ezo
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BEST
COPY AVAILAR {AsomAToRY {
Av
Vame:_Erre Race
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2
giReoqucesot .Date: __Myzeh T5 (Tv Prob No
| DAnualeysCt:opied; 2780
CSoadmeplNeo.Oster Taro ~ k TE 160
Dare Need:
-- | wt Hours Spent: E23
C--
PDeusrcproispstioonr: OSEECDD ly [[fio TSJesTEl oEnEFCeC20a3e F FC 20z6o oTC) 317] Please Call Requester If Delays Are Encountered.
wim |epee] DaonsitsConpusion car teruns) Sroyr --
T T [9 a2 t T do TTeh ras sso
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