Document Yj9Z3YEvzR4BxL6paGN8nRMyN
TOXICITY TO AQUATIC PLANTS
TEST SUBSTANCE
Identity: NN--eEtthyFlOpSeErfAllucoorhooocltoarneFMs-u3l4f2o2n.ami(d1oe-tOhcatnaonle;sumlafoynaamlsiodeb,eNr-eeftehryrle-d to as 1.12.2.3,3,4,4,5,5,6.6,1.7,8,8,8-heptadecafluoro-N-(2-hydroxyethyl)-,
CAS # 1691-99-2)
Remarks: Mjaimtietreidalwaitsearnsooflfu-bwilhiittyeo,fwtahixsymsaotleidrioaflupnrcohhairbiatcetderaizdeefdinpiutriivtey. The determination of the aquatic toxicity.
METHOD
M60e0t/h9o-7d8-F0o1l8l;owAeSdT:M-MEo-d3if5i.e2d3(DmroadfetlNeod.)
after those 2; OECD;
described by AG. Payne.
USEP--A
TGyLpPe: (tNeost type): Acute Static bioassay
YSpeeacriesst/uSdtyrapienr/fSourpmpelide:r:
1981 Freshwater
green
algae,
Selenastrum
capricomuturm.
AOnbatlayitniecdalfrmoomniUtSorEinPg:A--_ERALl,gaClorcevl.l,cOoruengtson(c(eJlullsy/m1l4),a1n9d81t)e.mperature.
EStxaptoisstuircael pMeertihodo:ds:4,7, 10E, Casndva1l4udeasyasnd 95% confidence limits were
calculated utilizing a linear regression model.
Test Condition Remarks:
Is1tn0ao-rdctaiuynlg-uoomlp:dtsiAtmlougcmakeciufnlrotocumurleauim7n-slduearvyeel-dootlfhdegsrpteorecenkseacnlulcgtaeueroef1xwa1e0srueffciueclsileesdn/tmaln.suimTnbhoeecruulosufemvoitfaobaglie7vetoa `asltgoaclkcceullltsuirne twhaesexdpeotneernmtiinaeldgrusoiwntgh aphhaesmeo.cyTthoemeintietarl (a3l3ga8l,0c0el0l cceollusn/tmii)n. the, AmlegdailuNmutprrioevnitdeMdeadlil ummin:erSatlerniulteriseynnttsheetsiscenatligaall fnourtrailegnatlmgerdoiwutmh.anTdhiaslsnoutsreirevnetd
as the diluent for all algal operations. The pH of this synthetic algal medium was
adjusted to 7.50.1 prior to use in assays.
Culture Flasks: Sterile culture flasks consisted of 250 mi Erlenmeyers
containing 50 ml synthetic algal nutrient medium.
Full-Scale Test (Definitive):
006420
Afllasdkesficnoitnitvaeinailgnagl5a0ssmaiyosfwteesrtesoclaurtriioend.ouTthiendterfiipnliitciatvee uassisnagy2c5o0nsmiistEerdloefnfmoeuyrer exposure periods: 4, 7, 10, and 14 days. T56h0e,f1o0ll0o0w,inagndN-1E8t0F0OmSgE/LAlwcoehroel,usLoetd 7t1o6i,niltoigataeritthhemiecxpcoosnucernetrsattuidoyns(i:nd1i8vi0d,ua3l20, weights). Three flasks containing 100% fresh algal nutrient medium plus algal cells comprised the nontreated controls. 0.5 mi Ethanol/ 1 liter algal nutrient medium was used as a carrier solvent.
Test Conditions: Aclolnatlagmailnoapteiroantiwiotnhs bwaecrteericaararinedd ootuhteurnadlegraea.seApltgiaclccounldtiutrieosnsweirneormdaeirnttoaianveodidin an environmental chamber under the following standard growth conditions:
Temperature: 232C (70-77F) Fluorescent illumination: 400
ft.
candles
10%
Free gas exchange: Continuous Platform Shaking, 100 10 rpm.
RESULTS
---
Tsoelsutbiinligtyenlidmeitdowfitthhe4t,es7t, m1a0t,earinadl.14E-CdsaoyccaallccuullaatteeddvEaClusesvawleurees ggrreeaatteerrtthhaann 1th8e00 mg/L (based on reduction in cell count).
CONCLUSIONS Calculated ECs valus indicate this material has insignificant toxicity to algae.
DATA QUALITY tReesltiasbuilbisttya:ncKeiiamnidscahctruaanlkimnega=su3.reTmhiesntsstuodfytlhaeckasmoinufnotrmoafttieosntosnubpsurtiatnycoef itnhe solution. Testing was conducted at approximately 5 orders of magnitude above the solubility limitofthe test substance.
REFERENCES 3MeMtTheocdhsniDceavleRleoppomretntSu--mNma-rEyt,FOMSuEltAil-cPohhaosle, ALlogta7l1A6s,sRaeypToersttNMuemtbheord,00N8e,w Project Number 9970030000, M. T. Elnabarawy, December 30, 1981.
OTHER
_--
006421
Submitter: 3M Company, Environmental Laboratory, P.O. Box 33331, St. Paul, Minnesota, 56133 Last changed: 5/18/00
006422
[Rp
TECHNICAL REPORT SUMMARY
12/30/81
0: TECHNICAL COMMUNICATIONS CENTER -- 201203 GUomrpeamretanst=taIfsreipsorthsornitaremtainobnoLoskiedensoof p7a6pe6r,(3s.a5n5d45t)wo copes TCC. See "Guidelinesfor Completion of Technic Report
HnEnvironmental Laboratory (EE & PC)
w Methods Development --- FM-3422, Lot 716
g053e5 l
9970030000
Multi-Phase Algal Assay Test Method
. Bacon/R. Bohon/E. Reiner/h. velter i. 7. Blnabara
008
16981
secuRmvD
FER
WL0o8m0,comaamia Roi Rumoriaion | RESTMTS>
rFoEan T--e--
ESGreEena ahlagtaee Generations
t"hSeelaelngaasltrgarmowctahprircesopromnusteunsof ovferreshswevaetrearl as they may be affected by exposure co the
fluorochemical FM-3422, Lot 716.
2. To develop an algal test protocol based on variable
exposure or recovery periods to the test compound.
BBE o6 nPC yDiv. | REPORT ASSTRACCTTTmormseromeiro mene ou Te Commo Con en He
In these triplicated four-phase assays, the initial
ewefrfeectassseonsseodrgaonviesrmssevfeorlallowignegnervaatriioends.expoTseusrtesrestuoltFsM-3ga4v2e2
an indication
populations.
of
Al
the
gal
possib
biomass
le
wa
effects on
s assessed
algal
quantitatively
>
in
terms of
response
algal
ECgg
cell-count
(mg/l) was
(No./ml),
calculated
and
the
median
growth
.
ptermadon [
006423
FPaHg-e34T2w2o/MTE December 30, 1981
coNcLusIoNs
1) McaotnesriidaelrsedpotsosebessiinngsiagnnifECiscgantglryeatpehrytothtaonxic1,0to00amlgg/ale aarnde aquatic plant species.
2)
iRnedsuucletds
niondgircaetaetder trheatducetxitoenndeidn
exposures (e.g., >7 algal cell counts.
days)
SUMMARY A four-phase algal assay was developed to evaluate algal growth craepsrpiocnosrenuotfuntheovferresshewvaetrearl ugneinceerlaltuiloanrsgarseentheaylgameay"bSeelaefnfaescttreudm baylgaelxpoassusraey etvoaltuhaetefdluaolrgoaclhemgircoawlt,h rFe-s3p4o2n2s.es dTuhriisngmulftoiuprhase gcroonwstehcutrievseponesxeposwuasremepaesruirodesd: in 4,CET7,TS 10Of, aan increase in Ablgiaolmass casalcaullgaatliocnellof-coEuCnsto's(No(.m/gm/ll)). werCeella-vceoruagnets roefsulttrsipluisceadtefdorsetthse of test culture flasks. vIanlutehsese(mtg/rli)plifcoartefdremsuhwlattieprhasgereeanlgaallgaaessa"ySse,lentahestrcuamlculated ECsg cthaeprimceodirannutugmr"owtahrerepsrpeosnesneted(ECisnoTmagb/lle) 1.folTlhoewsiengveaxlpuoessurreeprteosetnhte test material for 4, 7, 10, and 14 days.
INTRODUCTION
:
aIqnuatteisctinegnvitrhoenmpeonsts,iblueniceefflelcutlsarofalgcaheemiacrael rescuobmsmteanndceeds aosn athemodel S(yasqtueamticforpriemvaarlyuatpirnogductehres)i,nfaluqeunacteicofplacnhtesm,icaalnsd pohnytaolpglaalnkgtroonw.th
Tcoexlilc-dievfifseicotns daurreintgesttehde otenstg.rowiTnhgesealggarlowtchultruersepsonstehsatmauyndebre:goa) stimulatory, b) inhibitory and/or algicidal.
MATERIAL AND EXPERIMENTAL DESIGN Tlhiemitteedstsosluubbstialnictey i(nFMw-a34t2e2r, atLotamb7i16e)nt irsooamn toefmfp-ewrhaittueres.olidT,hewith bioassays performed on this material evaluated its potential arlegpaolrt.toxicity and the data generated form the basis of this TthheosetedsetscprriobteodcolbyuUtiSlEiPzAed- 6f0or0/9t-h7i8s-0s1t8u;dyASwTasM-Em-o3d5i.f2ie3d DarafftterNo. 2; OECD; A. G. Payne. (1), (2), (3), (4).
006424
PFaHg-e34T2h2r/eMeTS December 35, 1981
Test species:
aAlgbaaecte(rCihal-ofrroepehycceualet)u,re Soeflentahestrfurmeschawpartiecrorpnluatnukmtonoifc thgereoenrder c(hJluolyroc1o4,cca1l9e8s1).wasTohbetatiensetd aflrgoame aUre Sno-nE-EmRoPLt,ilAeCournvi.,celOlruelgaorn cceullltsu,re hwaavsingstotrheed apinpeatrheancdearkofata n4e9Cw. moon. The algal
Inoculun:
gAilvgeaea fsrtoamrtain7g-doapyt-iomludmsitnocokculcuumltulreevelwerofe gurseedenasalgianeoculum to
T2S5e0laeanays-t0ru1m8 cSaptroicckorneuutlutimre1 sufficient number of viable
ixns1u0r4edceltlhse/mplr.esenTchee oufsea of a 7 algal cells in the exponential
gcruolwttuhrephwaasse.detTehreminienditiuaslingalgaalhecmeolclytocomuentterin(3t3h8e,00s0tock
cells/ml).
Algal Nutrient Medium:
Maindnertaelsti(nignoarlggaaneic)wasstapnrdeapradrednuatsrieonuttlimneeddiumin fAorttaccuhlmteunrting#1. Tfhorisalnguatlriegnrtowtmhedainudm aplrsoovidseedrveadllasmintehrealdilnuuetnrtienftosr aelslsenatligaall oapdejruasttieodnst.o 7T.h5e+.1pHprOfiorthtios ussyenthientiacssaaylsg.al medium was
Culture Flasks:
.
cBaocmhpri2s5e0dmla tEersltenmfelaysekr. coCnutlatiunrieng fl5a0skmsl aonfd taelslt ostohleurtion g#2l.asswAaurteoclvaevreed spfeocamialplluygspreupseadredasasfldaesskcrcilboesudreisn, Atptearcmhimtetnetd free gaseous exchange to occur.
Full-Scale Test (Definitive):
.
uAslilngdef2i50nitmliveErlalegnamleyearssafylsaskwsereconctaarirnieidngou5t0 miln otfritpelsitcate ssoilmuutlitoann.eousTheexpdoesfuirneititveestsa:ssay4, co7,nsi1s0teadndof14 fdoauyrs.
T1h80e, fo32l0l,owi5n6g0, Fi1-,3040202,andLot1,870106 mlgo/glarwietrhemicusecdonctoentirnaittiioantse: the enuptorsiuernet mseedaiyu.m plTuhsecealgEarlatEceelTlsoitcioimnpirnigsed10t0he Envoensntrecaitoestd cfolnastkrsolsa.re oPurtolciendeudre inandAttsatcehpmsenftor #3p.reparation of algal test
006425
FPaHg-e34F2o2u/rMTE December 33, 1981
Test Conditions: cAlolndiatligoanls,opeinraotridoenrs twoereavociadrrcieodntaomutinautnidoenr waistehptibcacteria and oentvhierronamlegnate.al Aclhgaamlberculutnudreersthweerefomlalionwtinaginesdtanidnaradn growth conditions: Temperature - 23 + 29C (70-779F) Fluorescent illumination - 400 .C. + 10% SFhraekeinggas10e0xc+han1g0erp-m.Continuous Platform
Algal Biomass Monitoring: Tuhseingalgaallgalgrcoewltlh croeusnptosnse(cwealsls/amplp)r.aisePdrocqeudaunrteistatfiorvelaylgably biomass measurements are briefly described in Attachment #4. EuCtsiglizvianlgues3ManSdIXC9U5R%, coanfTiRdAeCncSeystleimmitfsorwerreegrecsaslicounlatmeoddels of experimental data.
TEST RESULTS
STheelenraesstulrtusm cofaprbiicoomranssutummeasuuserdemeinntsthoefsethsetudgireesenaraelgae
.
SCaelclufla-teedxpEClsana(mangdt/lo)arrevyaldueetsailienddicinatitnhge aaltgtaalchegdrowdtahtarsehsepeotnss.e to
dFaHy-s342a2r,e pLrotese7n16t,edfoilnloTwaibnlge e1.xposRuCrsep cpearlicoudlsateodf v4,alu7e,s10w,ereand 14
greater than to algae.
1,800
mg/l,
indicating
insignificant
toxic
effects
TpheerfodramteadrbeyporM.tedT. heErlenianbaraarwey baansdedR.onR.stRuodbiiedseadue.veloApcecdompaandnying data sheets comprise original data.
cIatlcsuhloautleddbeonnottheed btahsaits oafll thtehe irneiptoiraltedconEcCenvtarlautesionvsereof Fi-3422, Lot 716, in test solutions at the beginning of the bioassay.
006426
FPHag-e342F2i/veMTE December 30, 1981
REFERENCES (1) MSiellleenra,stWr.umE.C,apJr.icoC.rnuGrteuemneP,rinatnzd AT.lgaSlhirAosysaamya.Bott1l9e78.TestT:he
FUX.pS.eriEnnevnitraolndmeonstTaoln,PraoptpelcitciaotnioAng,encayn,d dCaotravalilnitse,rprOerteagotni.on. EPA-600/9-78-018. 125 Pp. (2) ACSoTnMd-uEc-t3i5n.g23T.oxic1i9t81y. TesPtrsopowsietdh FSrteasndhawradterPraancdticSealFtowrater Algae. Draft No. 2. (3) OEEfCfDectGsuiodneliBnieosticforSysTteesmtsi,ngTeosftCh2e0m1ic"aAllsgae(,198G1r)owtShectIinohnibi2,tion Test," Adopted May 12, 1981. (4) APlagyanle, ToA.xiGc.ityandandR. ItH.s HAaplpll,ica1t9i79o.n tAo MtehtehoSdafeftoyr MAsesaessusrmienngt of New Chemicals. ASTM STP #667, p. 171-180.
006427
TABLE 1
Algal Growth Response to
Based on RFeEMdC-us3cg4'a2st2i,o(nmgL/oIltn) C7(e11l)5l - Count(2)
Exposure (CDoanytsact)
4
>1,800
7
>1,800
10
>1,800
14
>1,800
(1) Method of ECsg calculation: 3M SIXCUR, a TRAC system for
regression models of experimental data.
(2) Algal cell-count (No./ml) measured in triplicated sets of
*
culture flasks.
x
.
006428
ATTACHMENT 1
NUTRIENT MEDIUM FOR FRESHWATER ALGAE
A. "(MPArCeROpNaUrTeRdIEsNeTpSarSaTtOsClKywSOiLtUhTdIOeNiSoni(CzOeNdCEwNaTtReArT.E)D) 1) 25.500 gn NaNO in 1 liter DI water. 2) 1.04 gnKHPO, in 1 liter DI water. 3) 12.159 gn MgClz 61y0 in | Titer DI water. 4) 16.700 gn MSO THO in 1 Titer DI water. 5) 4.410 gn CaCly 240 in 1 liter DI water. 6) 15.000 gn NaHCO3 in Titer DI water.
B. MICRONUTRIENTS STOCK SOLUTION (CONCENTRATED)
"(Co inamsb ingi len onee -lid ter stock mix.)
1) 185.5 mg HyBoy 2) 415.6 ngMnCl bip0 3) 3.27 mg 2c, 4) 1.43 mg CoCly 6Hy0 5) 0.01 mg cucl,
6) 7.26 mgNapHoOy: 2H20
7) 9 mg Fecly 8) 300 mg NagEDTA*2H20
;
.
PREPARATION OF SYNTHETIC ALGAL NUTRIENT MEDIUM - Add moinceromlnutorfieenatcsh msatcorcoknumtirxiepnetr s1tolciktersolouftidoenionpilzuesdownaetemrl; - sonicate and then filter through 0.22 um membrane; - adjust pH to 7.5 0.1;
- store in the dark at 4C.
006429
AERCHRENT 2 PREPARATION OF ALGAL CULTURE FLASKS All flasks used in maintaining and testing algae were made of
borosilicate glass (KIMAX).
- Flasks were brushed inside with a stiff bristle brush; ~ washewditwhitthapnwoant-eprh;osphate detergent (MICRO) and rinsed 3 times
=~ rinsed with a 10% HCl solution; = rinsed 3 times with tap water and 3 times with D.1, water;
- dried in an oven at 70C for 2 hours (placed inverted);
= and autoclaved with foam plugs inserted at 121C for 20 minutes.
006430
ATTACHMENT 3 PREPARATI0M OF ALGAL TEST FLASKS
ALGAL FLASKS conTRoL
ATGAL WEDIUR (m)) 3% Ng.s
1180 wll
: B20wfl| 3 Sow) | 4 1000wall
slecoul|
6
ABS
HBS Hes ug.s
M8S
EM {2 CoREoRnTS e.
+ TEST FATERTAL + ALGAL TNGCULUR = TOTAL
(m1) or
Oud (m)]
m)
.
+ LS
-
50
+q
+ |p + 28 + 50
+ 90
.
+ LS
+ LS + LS + |S
CLS
+
-
50
-
50
- 50
-
50
sw
-
50
7
+
+
-
50
1) Determine initial algal cell count in the stock culture.
2) Adjust pH of algal nutrient medium to 7.5 + 0.1 ml.
#
3) Make up `stock solutions with algal nutrient medium as desired.
A) Control flasks, preparations:
.
1) Add inoculum (volume predetermined); initial cell loading of 1 x 10 cells/ml;
2) Add algal nutrient medium to bring volume to 50ml; swirl flasks, and place
autoclaved foam plugs. 8) Test flasks:
1) Add algal nutrient medium;
2) Add test material (dissolved in algal medium if required);
3) swirl flasks;
4) Add algal inoculum;
:
006431
5) swirl flasks, and place autoclaved foam plugs.
c*e IoNSDIaVlIDURAnL,uoWnEI/GH1 TISmPRLACAELDSADLIRNEUCTTRLIYENITNTMOECDUILUTM,UURSEEFDLAASS,KSC,ARRIER, SO. LVENT,
=-HAeLMaAcvTIoNMcEcTuEuRu,m.CoVuonutusmE
33,354,306 N\,
PER Fas,
25;
(LxioY
23.8 338,00Q
cel ladX.
cens/ml.
=141.S8ad=i
ATTACHMENT 3 PREPARATION OF ALGAL TEST FLASKS
ALGAL FLASKS
CONTROL
) 2
"ALGAL MEDIUM (m1
N8.S
350wgf | NBS
4
!
TOTERTS ASA
KOT
2.
+ TEST MATERIA]
+ ALGAL INOCULUN =
(m!) or (maXsnmf
m1]
+ -- + .S
-
+
+
-
+
.
-
+ 28 LE -
+
Co
=
TOTAL (m1
50
50 50
50
os
5
+
+
-
50
6
.
.
- 50
7
+
+
-
50
1) Determine initial algal cell count in the stock culture.
2) Adjust pH of algal nutrient medium to 7.5 + 0.1 mi.
Es
3) Make up `stock solutions with algal nutrient medium as desired.
A) Control flasks, preparations:
1) Add inoculum (volume predetermined) ; initial cell loading of 1 x 10* cells/ml;
8)
2)
Test
Afdldaskaaslu:gtaolclanvuetdriefnotammeplduigusm. to
bring
volume
to
50ml;
swirl
flasks,
and
place
1) Add algal nutrient medium;
2) Add test material (dissolved in algal medium if required);
3) swirl flasks;
*
4) Add algal inoculum;
006432
5) Swirl flasks, and place autoclaved foam plugs.
*
-
HINeDmIaVcIqDrUoAmLeTWEeIrGHCToSunPtLsACE3D3,DI3R4E,C3TL0Y,
4I1N,TO25C5UR3T3U.R8E
FL3A3S8KS0,00
ceils/
0
-- ALgAs lowium NowePerFade. Ou. 1.5 fon 14
338,
ATTACHNEN4T
ALGALBIOKASSHONITORING
.
A. GRAVIMETRIC CELL DRY WEIGHT (1)
The fpiolrteersirzeec.ommended is Millipore type BD with an 0.6 micrometer The method is as fol lows:
= Ory filters for two hours at 70C in an oven;
- Cool lfeiaslttertswo ihnouarsdebseifcocraetorweicgohnitnagi;ning desiccant, for at - Filter a suitable measured aliquot of the culture under a
vacuum or pressure not to exceed 8psi;
- Rinse the filter funnel with D.1. water;
= Dry dtehesicfcialttoerr ftoorcotnwsotahnoturwseiagnhdtweaitg7h0.%, cool in a Basic instrument used: Analytical Balance; Mettler HE 30.
B. SPECTROPHOTOMETRIC DETERMINATION OF CHLOROPHYLL a (2)
Basic instrument used: Stectrophotameter; Bausch & Lonb, Spectronic 20.
C. IMPROVED NEUBAUER O.1 mm DEEP HEMOCYTOMETER COUNTING CHAMBER AND OPTICAL
MICROMETER (Used to measure diameter of a [gal HCV). (3) -.
v
Capricornutum Printz Algal Assay Bottle Test: Experimental (1). Miller, W. E., J. C. Greene, and T. Shiroyama. 1978. Selenastrum Design, Application, and Data Interpretation Protocol. Ecol. Res. Series EPA-600/9-78-018. Corvallis, Oregon. pp. 27. (2). APHA-AWWA-WPCF (1975). `Standard Methods for the Examination of
Water and Wastewater," (M. Franson, manag. ed.), l4th edition,
pp. 1030-1031. (3). Brite-Line (R), American Optical Corporation, Buffalo, New York.
006433
