-
'f f Icoo f T'fp'f f Icofo f iTo'xicTaF 1t«1'7'»»'4)«!i«i™̂ ";*K«-*-*" (•• riW •' -—&\+>i-fyJ~JL,|PBf̂ onTA|errcy
S&̂ &$&l$a$&!##»«̂ —-"̂
-
DOWTHERM*ĵ Al-HEAT|TRANSFER
TRANSFER
FLUID1TOJAQUATICIORGANISMS
CffDlliIfSciencei|Researcfi6wJChemical|Û *tAMidlandftMichigan
Fathtad i>r»5*»*«r>M>r~i-̂ ~r>iEpreraalai
9 S ccircuiatio
*°DOWTHE rtiptctivtiy*m*•" imsRi»n
-
INTRODUCTION!
mi blafJ 11 i " .i.-j ̂.̂ x̂*t. . . • ._ _ _. . w -.̂j.
•ngitheinonna;
• ' ' - - . - '
ni>r H
iWctlCTX?j£HTlms.ir-t*-?_«HK̂ Ei.*fc-j.i-*n*i.?»y J _, • .«FS--cLS&Bkfc.___- ._.J
-
PREPARATION* OF TEST ?tATERIAL
The teat samples of DOUTHERM A heat transfer fluid were
supplied by FPtS-TStD, 2020 Building.
The two samples cane froa Lot IMM06076. One represented
new DOWTHERM A heat transfer fluid as sold and the other
the sane lot number after 8 weeks of thermal degradation
at 399*F. A forced circulation test unit at 680 Building
was used in preparing the degraded sample, which had the
following characteristics!
1. By modified AST*M (distillation) method4 - 10% (vol/vol)high boilers
2. GC-Flame lonization /GC-Mass Speerrcmstry
Weight %
35.7 DOWTHERM A
12.9 GC-Recovorabl« Degradation Products
1*4 GC-Unxecoverabie Degradation Products*
100.0
From an analysis of the maas spectral data, the following
structures have beun proposed for the GC-rocovarabl« degra-
dation productsi
a"Hi -fh hoH ^rn" whi
-
-4-
Zsomers of this type:
biphenyl phenyl ethers -
diphenoxy biphenyls +
ter- and quater-phenyls
2-phenyldiphenyi ether
triphenylmethane
benzene
phenol
dibenzofuran
1,2-diphenylbensene
> 11.11
1.8%
112.91
The degraded DOWTHERM A heat transfer fluid is believed to
represent material from & typii.** .uscomer's operation* but
without contamination derived from chemicals other than•
DOWTHERM A.
METHODS
The static water acute toxicity tests followed the test methods
described in the U.S. EPA publication, "Methods for Acute Toxi-Tests with Fish, Macroinvertebrates, and Amphibians"7.
-
-5-
Fathead Minnow Toxicity Test
Fathead minnows (Piiaephales prone las Rafinesque) were accli-
matexi at 12*C to laboratory conditions for at least 10 days
prior to use. They were held in a 16-hour light/8-hour:Q
dark cycle. A synthetic diet was used to feed all fish
during the acclimation period. Fish were not fed during the
test. Test fish were placed in the bioassay vessel 24 hours
prior to addition of the test compound.
Tests were conducted by placing 8 liters of carbon filtered
Lake Huron water in each of 10 vessels, (8 treatment, 1 ^̂
vent control, and 1 untreated control), 22 cm deep x 24.5
en diameter round glass aquaria, adding fish, then aerating.
Stock solutions of the test compounds were prepared in ace-
tone. The maximum amount of acetone added to any aquaria
did not exceed 0.5 nL per liter of Lake Huron water. Before
addition of the test compound, aeration was stopped and the
test che-nical added, followed by C liters of water for mixing,
making a total volume of 10L. A refrigerated water bath main-
tained the temperature at 12€C + 1*C. Ten fish were exposed
to each concentration of compound. The fish were observed
daily and dead fish removed. 4fc
-
Daphnia Toxicity Test
acute invertebrate toxicity test consisted of exposing
daphnids, Daphnia inagna Straus, reared in our laboratory to
various concentrations of the material .in carbon filtered
Lake Huron water at a temperature of 20*C ̂ 1*C for 4ti
hours, with a 16-hour light/8-hour dark cycJ •:.
Stock solutions of the test compounds were preparHI using
acetone as a carrier solvent. The required amount of stock
solution was combined with sufficient carbcn filtered Lake
Huron water to make a final volume of 200 mL in each 250 mL
test beaker. A water control was set using carbon filtered '
Huron water. Because acetone was used in the stock
solutions, solvent controls were set containing the greatest
amount of solvent used in any toxicant concentration. The
amount of acetone was limited so that its concentration did
not exceed 0.5 mL/L.
Ten first instar daphnids weru added to each beaker and the
beakers set in a 20'C constant temperature incubator with
a 16-hour light/8-hour dark cycle. Three beakors were used
for each concentration and each control. Mortality data was
recorded at 24 and
-
Statistical Calculations
For each sot of mortality data, the LC values and their
confidence intervals are given. The LC10-50-90 are the esti-
mated concentrations of the test substance at which 10, 50,*
or 90% of the test organisms are dead at * specified time
interval. The LC values were calculated using Finney's methodaof probit analyses with a computer program.
RESULTS AND DISCUSSION
Two tests were run on undegraded DOWTHERM A heat transfer
fluid before data was obtained which fit Finney's probit
analysis program. The second test covered a narrower
range of concentrations than the first. The fathead minnow
LC data for degraded and undegradcd, run *2, DOWTHERM A
heat transfer fluid are presented in Table* I and II, re-
spectively. The 96-hour LC50 vaiuas for fathead minnows
were 7.1 mg/L (6.3-8.0} and 9.6 ,r.g. L (3.3-17.6) for degraded
and undegraded DOWTHERM A heat transfer fluid, respectively.
The 96-hour LC50 values are significantly different, p • 0.05.
The thermally degraded material is slightly more toxic to
fath«iad minnows than its undegradod counterpart. Ma3or
distress symptoms no tod during thu tt4t were a loss of body
equilibrium (fish swimming disoriented) and malanization
(dark-n-d body color, . ftR32 I
-
The daphnid toxicity test* were run three times because
mortalif • of the control organisms exceeded 10% in the first
two tests. Tables ZZZ and ZV present the daphnid LC data
for degraded and undegraded DOWTHERM A heat transfer fluid.
The 48-hour LC values for daphnids wure 1.27 mg/L (0.64-1.82)
and 0.72 mg/L (0.26-1.05) for degraded and undegraded DOWTHERM
heat transfer fluid, respectively. The 48-hour LC50 values
are not significantly different, p • 0.05.
Dennis C. DillResearch Biologist
APPROVED BY«
Howard C. AlexanderAquatic Biologist SpecialistEnvironmental Sciences ResearchHealth & Environmental SciencesDow Chemical U.S.A., 1702 BuildingMidland, MZ 48640 U.S.A.
AR32IH6
-
-13-
REFERENCES
1. Dow Products and Services for Industry, Farm, andHome. 1977. The Dow Chemical Company.
3. Sinunons, P. B., Branson, D. R., Moolenaar, R. J., andBailey, R. E. 1977. American Dyestuff Reporter.
-
TH2 DOW CHZLMICAt COMPAKY:
S:W W
• 1 0 L A M 0.• 1 C M 1 CAMmraan ——————— '
\WUIMI •—-——————————————————Analytical Laboratories
I. T. Takahashi and F. A. Blanchard*"" Determination of Uptake and Clearance Rates of
o«t«jMflrrh ?Q 1 QT+
^C-Diphenyl in »Rainbow Trout by Radiotracer Technioues,
ABSTRACT•
A radiotracer "C-diphenyl, baa been used by J. S. Bro«ierof the Waste Control Lab and the author* to obtain biocon-centration data for dipbenyl i& fish. Uptake and clearancedata at 1 ppb and 10 ppb exposure levels bave been obtainedfor muscle, remainders, and whole fish in a dynamic exper-iment. Normalized data for whole fish at an averageexposure of 1 ppb gave a clearance hall -life of 64 br anda maximum untflJCA ».tt.er BO hi*. A hlnr-ftrnr^n^T-af i nn taftru-of 1900 wa« calculated from this data.
K -' '"* ' ; • ••• •1 i
* • .
;* M '••-
ATTENTION
r-v. :--•••-: A; ̂ COPiEOlNMlDDiNQCRI. - . . - • • * • % , t • • j
* i . '.;
•: :- m '" :•! APR 2? 1983: "I
'• 1
, ; •— •—• . - - • • • - • - _
. i s ( tf e o TOOF | DEPARTMENT i ATTENTION OP
\
f DEPARTMENT
AR32UU8
-
March 29, 1974 - 2 -
DETERMINATION OF UPTAKE AND CLEARANCE RATES OF"•C-DIPHENYL IN RAINBOW TROUT BY RADIOTRACER TECHNIQUES
INTRODUCTION
A radiotracer study to obtain bioconcentration data fordiphenyl in fish was initiated as a joint project betweenJ. S. Brosier of Waste Control and the Analytical Labor-atory in order to evaluate potential environmentalproblems caused by Dow therm A (eutectic mixture of diphenyland diphenyloxide). Previously. Dean Branson of WasteControl had completed the diphenyloxide part of the study,using ^C-diphenyl oxide as a radiotracer.
Using similar radiotracer techniques, with ' ""C-diphenyl asthe radiotracer, bioconcentration data were obtained fordiphenyl.
In experiments using a radiotracer, the purity a'nd valid-ity of the tracer plays an important part. The ! l*C-dJphenyl,Lot no. 5109:10 f-2, was synthesized »•• Mar lone Wass ofAg-Organic Research. This tracer had a specific activityof 3.937 mCl/mroole. Since it had been stored in benzenesolution for one year, its purity was redeterraJned by thin-layer chromatography and liquid scintillation counting byM. Wass. Two solvent systems, hexane and carbon totra-chloride, on silica gel 254F plates showed the radiopurityof diphenyl- 1%C was 98.7%. Analytical data were alsoobtained on a standard sample of unlabelled diphenyl re-ceived from the Halogens Research Laboratory. Thin-luyorchromatography in the same solvent systems showed only oneUV detectable spot. An infra red scan by R. A. Nyquist(AL 62-213) and gas chromatographic analysis by Tom Petersshowed no obvious impurities in the unlabelled diphenyl.
EXPERIMENTAL
Preparation of the Tracer
The ^C-diphenyl at a sp. act. of 3.937 mCi/mmolo was re-ceived in a benzene solution with a concentration of 13.708mg/50 ml. 39.4 ml of the solution (10.80 mg diphenyl) wa«placed in a flask and mixed with 12.04 ms of unlabolleddiphenyl. A Rinco evaporator wa« used to remove the benzene95.8% of the radioactivity was found in the flask and 1.6%was Found in the trap to the vacuum line. The specificactivity of the carrier diluted radiotracer was determinedby liquid scintillation counting to ho 2.68 x 10*1 dpm/w
fiR321'i»lt9
-
March 29, 1974 -3-
An acetone solution (100 »1) containing 218.7 tig of diphenyl-**C per ml vac prepared froot the diluted radiotracer. Anotherdilution with acetone was made to obtain a * ̂C-diphenylconcentration of 4.66 wg/ml.
Fish Exposure and Clearance by J. 8. Brosier of Waste Control
A continuously metering, diluter system was used by Sam Brosierof Waste Control to maintain the concentration of • %C-diphenylin the exposure tanks. A liter of the acetone solution of**C-diphenyl (4.66 ug/ml) was added to th« metering reservoir.
A thousand-fold water dilution by the diluter gave an exposurelevel of 10 ppb in the first exposure tank. A subsequent 10-folddilution gave an exposure level of 1 ppb in the second exposuretank. Thus exposure levels of 10 ppb and 1 ppb could be main-tained simultaneously. This equipment is described in the reporton 1%C-dlphenyloxid«.
Forty fish were exposed at 1 ppb, another 40 at 10 ppb, and4 were kept as unexposed controls. Four fish were removedfrom each tank at 6 hr, 12 hr, 24 hr, 48 hr, 96 hr. Water wasalso sampled at these times.
The fish were filleted and separated into flesh and remainders/and frozen until they could be radioassayed. The water wassampled by pipetting 5 ml aliquots directly into 15 ml ofBray's scintlllator. These counting samples were stored Ina refrigerator until radioassayed.
After 96 hr of exposure, the fish were cleared in continuouslychanging fresh water. At clearing times of 6 hr, 12 hr, 24 hr,48 hr, and 96 hr, 4 fish were removed from each tank. Theclearance water was not sampled.
Radloassay of the Water
The l *C-diphenyl was determined by direct counting of the 5 mlwater aliquots by liquid scintillation counting in a PackardTriCarb 3324 liquid scintillation spectrometer.
Radloassay' of the Fish
All fish flesh and remainders were homogenized in a Virtusgrinder. All the fish samples except the uptake flesh sampleswere combusted in a Harvey Oxidizer.v Duplicate aliquots of00 to 200 mg were combusted. The ̂ CÔ formed was trapped in
flR32U*50
-
March 29, 1974 -4-
an «thanolamine/2-methoxyethanol (70/30) solution, mixed witha premixed toluene-xylene liquid scintaillation fluor (Econoflor), and counted in a Packard Tri-Carb liquid scintillationspectrometer, model 3380. The uniformity in quenching of thesamples allowed us to correct for the quenching by a representa-tive standards technique. The combustion efficiency wasobtained by directly counting and combusting aliquots of astandard solution of diphenyl-Cl% in acetone-water. An averagecombustion efficiency of 78.5% WAS found for the diphenyl. Th«ng of diphenyl-l *»C in the fish were calculated from the radio-activity found and the specific activity of the diphenyl-I%C.
The uptake flesh samples (100 to 200 mg) w«r« solubilized in2 ml of Soluene-100 by shaking the samples overnight at roomtemperature. The solubilized samples were mixed with toluenc-ethanol liquid scintillation counting fluid and counted witha Packard Tri-Carb liquid scintillation spectrometer model3380. An external source channels ratio method was us*d tocorrect for quenching in the samples. The wg of ^C-dipbenylwas calculated from its sp. activity.
The ppm diphenyl-1 %C in whole fish was calculated from theweights of flesh and remainders, and their respective radioassays*
RESULTS
All analytical data on fish and water are tabulated in Table 1,2, 3, and 4. The uptake and clearance data at 1 ppb exposureis plotted in Figure 1. The data at 10 ppb exposure areplotted in Figure 2. The composite uptake and clearance profilefor diphenyl is plotted in Figure 3. The diphenyl concentrationin the water, C^, is the accumulated exposure level at anygiven uptake *••*"* and was calculated from the actual concentrationfound.
From the data in Figure 3, the clearance half-life for diphenyl,t1/2, equals 64 hr. The bioconcentration factor, BCF, equals1900. Tne aodel appears to be a one compartment model. Thecalculations are on the following page.
AUTHORS
flR32ll»5
-
March 29, 1974 -5-
•. 0.505 x 10*
- 160-06 - 64 hrClearance ̂ 1/2 ~" *w—»w — w wk« - .693/64 - 0.0108 far"
^ff - k» C -k, CfHt^ w x
Let Cf/Cv - y
dCf - CVdy .
+ k2y
(0.10-.00)10» + o.0138(0. 10) (10>) - (0.021)10*
- '021_(10>) .- — 6. *
ki - 21 hr~lk2 - 0.0108 hr"1BCF - 1900
Note: The approximation here is that the tangent to thecurve at the first data point is coincident withthe straight line from the origin to that point.
AR32U52
-
m05
Is3*A
as
'iih Analysis
8.0b
•4>o
4*a:
iO,»asU
OS
1«k«HJ
«MB
Am ••H Otu Z
••w"r"fl4*4.
J
il
!
-
toentrH
OOcn 01
oAit*
H» M M MCB fc> erf u en CD «j en
A to ea «»it* ui «a A
M
** ** ** Cto m «j J,,
»- M H* 0 0 O.M MOMOVOCD-JMfcJ
M »- O MM 0 *J IU
M
0en
H* fj4O 0* »-• Ok
o ^ *• too *• •*• in
O O O O0* «•
-
t — fcf*
to
OB OB OB OB•o %i A en
erf A o £
.O OOOMMOO
0 0 M O4k 40 4k flD
OUl»fn
* ̂ ? P*• " w 3tO 40 W **
•— MMfOAA4k4k•kUjeo^iuiencrfMTtMWtOAMMM
- M Cn 4k .•) to VI MO O to M
J3
M M M•>! 4k O CO
eo o 4k enerf en o* M
O O O• * •M erf Men irf oerf o en
MOD
•
CD %J «J «-O 40 OB *J
A • A 4k 60
OOMMOOMM
en A eo o*OB en o en
MM
A «kl in 4O
irf M 4? «
OOOOOOOO
in et w cc M »••«/*
o o o o
91 o to wKl M to en0t4k-IVM M M MM erf o eoto A 4k enen o M en
o o o o
M I* tft *O»^_ A^ .̂ kB
f
0-4
•J
2?
̂ «j u ef
to OB en 4v
O
MAO
'
en en en enA en 4k erf
*J 00 «*l 4k
A 0 *J 0ea to o»
SBHsSSSSM «J CD M
•a to oo enOB M erf en
JrftOMMOtO o *• i•
3CMi
OMi
-Jo'
O?
•0
£rr
O
3 MB
e»ftt
•jt•*.•
4j»•r
L*•*H»»*J
?M<•H-•
i
1jr
•n>K^
H»cT:K
RScaGg
f
-
IBm04P*o«H
SccMftkl
"i
SAC
Hago*a
*
«H
fH1«•G
I•»1)••ii
1'iiS
gO
«4-•J
XIat
ic1tc.
I
E
«C
ia^••«u
D•4•>1-10
4Uu0B
•
b1
1»»
IU
kj«»,
*•
t
k
H)
M•>
02
O£
U
fi•H
«H 00 d Oo
o *r to e*d A A 4O
•H O O O
*4 e*̂ ̂ 5 C) C9 C9 C9 Ĉ
•
m4Od
•>
r» A ft OBOf t̂ fl €̂ $ *̂ fl
5 A r» d. «n «» r*2J « e» r»
r* OB A o
.
ft r+HO
S
eorH*•H
or r* m toAd O «H
O HI «H «H
QD A P» «H
O «•» 40 A
O
CD «H «0 •»* A r» o
otoodCD.nom-4
a a 2 . 2-A
m
•"•I ^̂ IO Ĉm A eo f«* to n VP
m w «» eo eo «r tn
S S 5 ^• • . .33- en
m %o fk eoCM d d d
mm
O Amm
r*
eo0d
•H n A OO r» «e» enr*. o »-«
-
toA
S
A en• •MiJ
. . 4k CO
A•irfM
M M M Hen ui en *M M 0 tfi
•
*» o *• ^in 1. *a *j3 M *
AO enM enoMUiM
O • irf M• irf • •A A M 4kCO irf 4» OOB M
M• • "
to
s - rjM J M i>l
rr ccce esss*?-- MAtV. O.AA02S01°* AenM4k 40A-JOB***1
M MA erf en toirf M o enM 4k OB M
•» mM • •M M « J• OB irfOM
M M M40 M «0 **» in «« ^M A trf W
erf. A W O>
MA in entO irf CD MM «J M A
en4*
enoto
00
f?en en• •4k erf
. OB «1
.
- en. •erf
M M M MOk Ok Ok Ok4k irf M M
4k -U M g
•** *»> w r,*j M w ;3
O4»inenM4ktoto
M M M M,_. • . •S *J 0 M2 0 40 A"* en oo OkM.«*1
ill
in *i A gA 40 A "in irf -i g
• •
OBtOAAAAtOtOMvjtoeoentrfotovoMen*dvoenenA
ea A en 40»0 40 4k illen M «J M
-J•«0A
M M M MO in M M4k irf M MM in O 40
en o> irf enOf irf iH 4O4k 40 W irf«J 4k «U M
•
M
••3Ml\!1
fi» 11M
•O 1•O 1tr i
as*?\
T,Ml
0Ml•
1
X9)
O90«*
*. «•O1
«ft
OIt«
1
5f»krf>
>»•.̂C•.*.•
-
KMCQP*cw
5mMIM
1o.Moa*oMO
1
.
„,̂
rH
A«Hfe
A•Hft,
r Analysis
[ss
1
D
X*3B
Q.
o:u
«
O»
IMO
«M3C
0
J3.a
i
to
m
CD r4 A . **tO *H 0 A
c*t d d oC* rH rH m
0 d iH 0rH rH rH
O4omeodd«neo
r* to to *e»
o
A *r A r»rH * co r* r*- co m
o o o o oo
n A A0 m r»
A 0 rHoo en en
AWrH
AR321U58