Document rBx2rBJmj78dVOBVbGdwy8J0v
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Pobnwiy 2T, 1957
Dr. 1'. r*. CinmiAB Do.ir-rrcD Diviuicn iivikJun I.'o. 33 i;.ltioaaX Ca-tU Sa<;istar l)yton !> Ohio
Oaor Dr. Vhcviao:
fittac'icJ is a photoncac oi eho oci^inal paper o Or. Jeneeii In Sweden, rotating to polychloriiiBtod biphenyls, X viU. Je> happy no have yor iucms a fter
yon ron'l .It.
ha aVt.r mo td> d section on torieeloq'/ is oonoornoo, it : Is tree <lnSt :`c!iloj;ii<niP.'nn(| live*- trtnule. eon result p-rf-i l"m> dears. ' -T/l'ethov"or* not t>ua is at oil xolov.uit
to nretil 7tru.ttlo* o:aiotirr in human iat in, ol colirso, nn oncixoly oifiorent ctueutimi.
At any rate. X beliovo before \ vorry about, t!io
toxieoloqioal part OX the problem, wo abould rattlo
tho analytical part, '
Sincerely,
KT.l'./lu att.
R. Eimtt Rally, n. P. Kedical Dlrcobox
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003114*
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DEFENDANT'S EXHIBIT
mmeu 2000087
MONSFOXOOOOOO88
WATER PCB-SD0000054848
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Hi* unman, Indite and gontlBnou* 3 honor to our Srittieh $e*I win try to bold this iet in
BngHab*
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As the tills of ihie loeturo states, 1 m todey going to toll about
tho fliMOTory ot some hitherto unobserved chlorinated Xldmouiloap
haying 19 eight chlorine in the aolceole nd tool'll residue bbo*
iysie# Tba ohonloel none of poiyohlorinotod Ufoajla ( Zb the following
colled KEB) to got fcnlliar with JOB I sill start tdtb tho ohosdfitxy
and esdo6logl
**
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Pborolotry *
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fbe Bain*charaotftrlatl of PCS Is 1`* SChetrvsry high stability* Afc m
.|
cxcapletbey can bo boiled-with nltrln noil without being floetrctfed*
'
2* Shop are hardly metabolleefl In living organ!ea 3 If oro thnn 4 ohlori&e Brelpcrebseat thoy edre non Inflau&le* IV 1b dewr that those
throe cb&ra^&xirfcles does It easy to understand 'tint when they hnrvo
-
entoroj^th* giving organim*- the will have' a loo porslntattce But It lo* difficult to ta&Inin hoe they find their my into* the living organics* . *
One thing soerne to he elenr, they don't ooae frou agricultural use,
*-
hut freu a technical. one and viOst probable it doses to the nature Via
,
waotee that are tried to be burnt up, becaana then re have then, at .onto *
in the air,* heoanoe of their son ioSlisM^ility* / '
:
ftOXtCOlOgj * * . . .
*
)
*
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The PCS were introduced la 1323-end an early aa t936 yonce.and Alden
j t j
reported that 23 -out of 24 aan eaployod in unnufftoturisd of PCB suffered
iron a am fora' eruption of the a)dn`JLc&* did sot appear until ( to
0 nentbo after the notorial who first'weed* In 1937 Brisker reported
g
that rote expooed t? chlorinated blpbeayia ia eonee&tnatioa of appfoxl- * * natly 1 sg/n^ for 1( bouro e day for ( rreofc` chosdd' dosage of tho lireir* g
After that tins the allotted uonoentratl&a of Ks in air ls.0,3 ngA , (For BBt the ease rolue 1* 0*3 * 1 *^a?) the sane authors finished
^
tbeir experiment* in 193Br and related that those eottpou&id have on Injourlovo effect, Manifested solely in tha liyer^CULorlnntod bipofcnjrloJ 0 appeiirrf to.be'-.tberaoet inieurlono efclerl&hied. eoipppuadp of all testod* j s
PIIGNCR 2000088
MONSFOX00000089
WATER PCB-SD0000054849
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Brtwiburg, Heyer esi Sstoh 19J9 repeto.d thet MB esd
.
Hapbfaleaca an biased Tor toe death r three yeunTyotoere,' iT'tbTt^
prognant vosen ua persons sb have at any tine bad tuf liver lMtws
or# particularly euepectlble*
.
Vedsl, Boiler and Benton gars 1942 untoals VCB inclosing odateirtroMon by inhalation, Insertion eng tots ebeerittcn. Mstaaododi ---- ** to* viscera shoved JmjiOrteiA toxin'attest iy to too eMl nil liver,
and to* dogenorrtltDO effect* to toe liver vs eoeentielly toe eene uhat-
ever veo'too netood tor toe atoinletraticm, partook (19K/ feoad as on
oecnpetlonal polooa to too eleptrieeX ioJuoiry, stood totxa tat ptaf
ehlerobljbecgrl esnsee Tolllcttlltie, eoaode, pyedoral* oat other
nttertlenoe and that toe principal trade offset is Tatty degeneration
of toe liver#.
*
Biller C1944} injected 99 ag JCB (4 end 5 chlorine) enbeontoneouoly in I'
52 guluoa- pigs. Bight to ton day* after inj*etiei fat droplet* were
* 1
noted ia tbo liver. oello, and after 15 days they were prteent in BCderoto wr very large nuaberao Rabbits and rets'were also t*etad la thlo inmtl gbtlwr M well a* the BCD vao adnlnistated bath sontlnoaalyr eubuatin
ouoly er ingested la .the food* In the feeding cxperlnsnt 0 guinea plge received i .docae of 09 g of the chlorinated biphenyl 1 weak apart* Death occurred in 11 to 29 &ay*
.
1
finally Mo Xaughlln 195$ repertei a Bathed to toot the cbejrfoel toxicity
and teratogenic effect by injection into the yolk m d tortile agge prior to inoubatlon* 3*CB wee found between the eight ooBpotmde anong 100 .tooted having the highest order of toxicity* Vo batch vae found at a level of 25 ng p egg* it a level et JO' ng par egg> one cMck hatebei oat of 20 injected eggs, but died * days later* Son* eabryoB which were cmnlned- after they died, showed weak'defornltlee (often a abort upper brak}
and growth retardation* Dead acetate resulted &e an example -la no hatch *
at a level of l ug pox egg* Autopsy of tbo deed eabyoe hove above* extea
slvc brain daa&ge* Hercarlo chloride ehovod no hatch evon at a level of *
0*5 ng per egg*
v
ia*the analytical ohenletry i* a pronounced service sedenoe X have boon in contact vltb nojqy scientists firca ether fields daring the work with residue analysis, and X have always found this contact vary stlwilating for ay own work* This do*-operation often deoanda that v taro talking the a Sana solentlflo language* Secauso of thia need X will today try to glvo a lecture In lew level analytical cheaietry for biologist^ lllnstroted by the reoidue analyel* of polyohlovinated biphenyl** She loeturc will be divided in the following three eub-divlslenet
FHGNCR 2000089
MONSFOX00000090
WATER PCB-SD0000054850
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1i Chenlvtry Dl PCS end
tocinology.
2* Anelytlcol methods for residue analysis u4 proof of etructurpo*
3 'Bebwiwar of MS In nature9 ditterenoies Is metabolising taXa
ot ibo TCB components, potenwrtit An as ecological eerie, ce-
oontretlim levels an& azasplss of emaplee otoicto hare Woo proved \a ecu.,
tain pco
*
A residue analysis eon W divided imi
\'
' Ir-Eartraetlon of the peatloidea Iroa the blologlsal' material,
* followed by a careful elesnlng^p to tole away intsrferriag
aubatanoest most often fats*
>9
St Identification eaalytrle toy naan of gae chromatography* Thin-
r ' layer ohronutpgraptoy end mass epaotxomatry*
9* Quantitative analysis*
'
*
. *
At an eaolegSeal laboratory In SUsaiuelet in Stockholm 1-2 $ ot a sasplo
is out out of too biological material and transferred into a weighed and
eartfully cleaned tost tube, cad stored at &0* until analysis. Snailor
o samples hare boon tised, ndn* 9 ag of body fat, end with dry materials ouch
as hair, feathers, pine needles 100 ng arc tniffieleat to reaob the dosirad
10 ng/g IsyoI in residue analysis* In cases of water proofs ) 1 is used `
for reaching the 10 pt/g* lertl*
Bl{bopog) In order to facilitate conplsts xtxiotion of the fatty smtsriels from
the biological sample, the doulbe amount ot finely powdered, anhydrous
* aagneeiuB sulphate is added to* tbs ssapling tube, and the Whole ls'tooB-
gsnlscd with an lasertahlw homcganlssr* ?be resulting powder la transferred
into a special Sexblot extractor. After'i hours of extraction the solvent
le OTspontsd, leaving the rat la a testall weighed tost tub# at the bottom t
'Box.-tubt) of the extractor, vtoie fat is Altoeolycd in nettylene chloride in such a
way that 100 ul (o,1 nl) costBin 20 sg pf- fat*
'
Jhe 100 ul solution is now transferred to n little object glass, 5n7 on,
* covered with a siiloagel layer 1 ns thick, in order to foa a line Ot7 cn
from one end of the slide, Inserting this thin-layer plate Into a vessel ,
the bottom of which is covered toy a few m of methylene chloride, the
3 olvout will be sucked up in the dry bye* of sUlcagel, and at least reach the upper end of the plate. 5*be faot is that the fat has a greator
affitdty to, the powder on the plate than the chlorinated hydroearboo
have* -- and we get a separation. tfbe fat toeing more polar than the
chlorinated hydrocarbons will never go laager than 2 cm before ttoa
. .' '
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FHGNCR 2000090
MONSFDX00000091
WATER PCB-SD0000054851
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eolVont boaeho* the upper part of the gloss.
ke front of the fat appear quite vioiblo against a lop, otA with the
aSA of a ra*or blado the sone above the fat la transferred to the elution
tube a*4 tt# chlorinated Vlooidoe absorbed on tbe ponder can mv be
eluted hy obo ul of ether* The ponoontratioa la rafflsisst fear detection of tho ohlorinotod hydrocarbons Ana to tho 10**^ g level*
tbo nest top in tho analytical procedure oonooxns tho separation of the
different Chlorl&atod tyilrpoarbone that tbs oasple uj centals. As a
patter of foot, thin le a broubloscme tadf Zt le easy to ortlsate nhot
io not present, hut acre difficult to nay exactly ena la present* 9
nuffor Area tho negative demoastrstlps, a will he ebon lator*,
At first n ion vordo about the separation of the aoapaianta preeost in the
sample. and their visualisation-
,
9be separation io aeooapllahoA by seas of a gas chromatograph fitted to a
detector that transfers Its inpulse to a recorder*
'*
`
fbo syeton io shortly ^teoorihefii
,
A spirally formed glass tube with an lanor diameter of 2 an and about 2
n in ietigth is filled up Vy a support, covered with an thin layer of an oil* The tube is heated in the ohronntogr&ph to about 200* Through the
tube a streea of nitrogen toaliaeusly fallows. When about 10 ul
ef
1 si) of the purified sample ia Injected Into the tube, the oompo&oata of
the owayle will bo erraporlsed and go forward through the colutm with the
gag strona* As the constituents havo different affinity to the ooluna
filling they will pose tho eelu&n with different speed end.it will tele
different tine for then'to roach the dotoetor at tho other end of the
glass tuba* IS the temperature end the nitrogen flow are hold constant
thin tine| tho retention tine, has a speoifiowalftb for a certain ew*peuafi*
fhlo le true, but unfortunately It Is also a fact that two ooapononte can
have the-wane retention tine* this lexeme of the bigger problems in gae
efearenatogtaphle analysis of xmhnowm seopXeB, ao mill soon bo obvious
So take It possible to estimate tho rotontion time it Is ttooesoery to
visualise the chlorinated hydrocarbons- For that purpose nor* or lose spooifio detectors cere used- She doteotox moot often,used in postlddo
analysis is the so Dolled elootxen oapturo detector, which oen detect down to one pioograa (** 10*"1 g of linden)* Dnfortunatoly thle detector Is not
spooifio for chlorine, bn gives answer oleo for ootygenoontalsing oebpeuade* She response hove Is such lower but eon bo counterbalanced If tho concen
tration of the oxygon containing
la much higher*
0281023
Tho principle for tho electron capture detector is shortly*
At the end of the gao ohromatogxnphie tube is placed 'a little tube conalar a roll mAb %*t tliuilwi tiitido. Thle in ae-redlaart* The o--
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PHGNCR 2000091 MQNSFOX00000092
WATER PCB-SD0000054852
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partiolee oxo roarting with,-the nitrogen Boleculee cowdag trxm tbo oolww.
Tb*a we got
+ -- 9** <-
Over the' detector a* have a tension
of 90 Volt end by mean of the electrons wo will get eoaataat electrical
ourreat over the detector* Jhie Mealing current le traaefvrrwd to a .
oao^&V recorder ae a constant beeollne* bhon now a ehlerimted J^drcarbon .
locvoo tbo ocluna thio cospound boo a hidk affinity t* the eleatreac end
thie scans that the cnount of electrons will
and they will
diminish proportionally to the enewt of chlorine# fhe electrical current
mil also dfatnlab and this is noted as a pea* on tbs recorder. fbe area
of tbo peak will be proportional to the mount o substance la the ea&ple*
By roan of a standard Infection it te now poaalblo to towpare the retca-
tlon tlno and the aroa of an unknown ooaponent with the retention tine
and area of tbo known standard. As said before this dotootor la not epeal~
"tflo for thltt&o but anyhow very useful^ beoauee of its Ugh sensitivity*
She ayeten described has, aa we bare Been, two dieadvantagoo:
1* {two different cowpounfls can hare the cane retention tine and bo
dotooted-'es oho peak*
.*
.,
2. A rcglctrated peak doss net heed to be chlorinated 1 beoauee the
detector is not specific*
If the-Maple le injected in two different colusne edtb different chemical
proportion vo have inoreasel' the chance for. a good oeparatlon. If t*o
compounds have the eaae retention tine on one Colton .they way not hew it
on another. Vbcn a result ecene doubtful# * If the oospeund being
f
reepo&elble for a certain peak contains chlorine or not - It is possible
to ooac.cntrete the sample and analyse ,-lt 00 a less sensitive detector `
such ae the Blcrodouaetarlc one, which is epaolfie for ehlorinc* She
compound`le burned in a furnace end toll generated chlorine titrated
directly*
.
le le ooen iron the two last aestioued possibilities it la anyhow possible
to get a rather high degree of certainty in residue analysis, but it to >
a rather tine-oonsumelng woxk.Vhcn using this toothed Juab described, we
j very often found that eany chrouatogrcoa fron Vesldne aaalyols of uoat carefully purified samples still contain a large mnber of peeks, Uayy I, of these hove retention tlffee that do not agree with any known chlorinated
pesticide*, or their netabcllte** thle chronatograa con serve as an
,
i exanple* It was obtained by Tealdue analysis of a owa-sagis-found dead
g
I in the archipelago of Stockholm. In the range of the known peaks, thorc arc so bony unldc&tlfiod that there aloo mint be an obvious rink of the * known peaks to be covered by unknown ones.
jf th4n mark in fmmr) 1w. ik. r.turt*<l
nf *
P0GNCR 2000Q$2 MONSFOXOOOOQ093
WATER PCB-SD0000054853
1tltublve bQ&!^J,p ISUt b bMUtikt ihto ^UO0\ 1j tke paweont inrrocti-
J
BKtiwi it 1b shown that most of th unknown peak of cfcroecUgraae at
rcoiduo snalyeia of chlorinated pestloldee are do* to polychlorinated
biphenyls.
.
'
I will ebow & chromatogram of human fat anulycod aim celled 6? $6
oolum9 the Boot often used type in pesticide analyses. Early retention
k
tints were In agreement with VBZ, XOTep end PKpp. Sect elide shove the
oane staple analysed on a Q?_1 column. lev the fNM 2 E peaks have
*
divided Into 4 penis,and two of then ere still In agreement with TOttpp ' end op#v the two now were unknown.
Xoglo&lly, thoee unknown oonponente wore nt first thought to he eetabolltos
of the insectioldem. Against that spoke that uoigtber treatment nor
ooseonbrated eulfurlo add In other, fhif treatment nade it rather, sure
that the compound* did not contain o^gen. in Sweden residues of orf'cnio nc?
oury have Veen Investigated rather intensively In the Swedish fauna,
*
As those oompouade fire very high responses to the electron capture detec
tor it was aloe layeati gated if the unknown peaks could havo a mercuric
origin.
*
It was found that the water-ecological eeriee had high residues of both
Boroury (Tfeetemark,Jehnele) and the unknown enee, when the sane Indivi
duale were analysed. Anyhow, the pheasant suffering nest from mercury poisoning only contained low level& of electron capturing ooBpounds and
theso belongod
*
'
to the normal Ineectisldee. Therefore tha unknown oould hardly he mereu-
rials or .metabolite# of 'then*
-
.
Ae the eagle eeraple giving the chromatogram cfrova in fig. Ip. oould Vo' estimated to oentaln 2DT and DEB up td 1> g/kg in extractable fat, the amount of unknown compounds aloe- were suggested to Vc in the eeme range, and teha sufficiently high to do a ran on the ooaoblned gas chromatograph - * pass spectrometer. If this could be done ewcecosdulAy it would bo possible' to get very Important informations about the ehemleel mature of the unknown, for ex. the x&olekular weight numbers of chlorine etc. This method le up to now the method giving the highest degree ox oprtsiaty In the low level analytical chemistry, amounts of 100 ng substance being enough.
6JVT>?0
As thio method for Identification at totally unknown residues surely will bo very important In the future (when fex. a biologist has found that fishes In a river die) It may^Joeeible by mean of this method to
find out craotly what oempovnds are responsible for the death# Tor this reason, I will go into come details with thLs method. In the actual oaoo vre took tha extract Croa 20 mg onslo end cosoeatrated
t
D it as tmoh Be
and wda AA ln>eotion OQ ib |U ebltaStO^^
oo&bla4 with the nee* speotroaeter* frba result iu ttt efcrewatogrftn
shown on the nwrt olldo. Every tine the recorder rtwi4 that a eooptmd
in leaving the column, the effluent la led to the ut apeotrcaaotor* Bov
Juet o fen vorde about the naan epee,
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fh seleculee leaving the coIubb are bonded with electron* at 7 Vo have do* got the wolooule poaitlve chargedt hoi with the in* woe a* before* ? This H* la.aoeolorated In a vnpuua and will then get a Jdaotlo energi .
whero " i the speed* Next cone* the nagaetle field that
trios to head the direction of the aolaeulo* Shis
will he
big for. a eoall ooleotilo and lead tor
If m have a sieve la the other end m orb dlreotljr read the eoleoular weight* Added to this parent solecula M+ a* will alao get addition In* formations, beetofoe of the fact that H+ toy not be etbble, a part of thon will be broken down botoro the; reach the alev* In tba other end* ^
?,<a* M vm **m - coij
'
Maes epeotrograae from the different unknown peaks In the eagle Mpl* as *
shown* The &aes numbers equal to the molecular weights of the uakaewno
could be read to 426>92 >58* >24* jCartonlehlnglyf the molecular Alffereiteee were constantly >4 wane unite* Shi* difference ahowo a fwllarity
la origin of the unknown* How the fact la that ohlorlno oxalate aa a
mixture of two leetopaa with atom wolghto 5$ and >7 in proportion 75i25 If
the moloeule haa'oae chlorine, thia will give two molecule peeks* one tor
. C1^ end one for Ol^j* If there are two ohlorlno wa hare the possibility .
of one with only
one with both 01^^ and >7 and one with 2 81 end
therefore
*
0glt20
PHGNCR 2000094 MONSFOXOOOOOQ95
WATER PCB-SD0000054855
nlatioQ ol ,tbt,,p6(ki iovuiS pa thv dlf^wpat mm iyio wri.
Volccular weight.?'-*?. ^ *
35 B
426
Chlorine content.;; * 5 ~ . S
70
An ttplttatioa pf
taoUluity of th compounds mb bs given if one
substance Is built from tiu? former by eubetltvtlng m hydrogen with
oblorlne
.
'
* 01.,2
B0. 1 JCC1
ft>tm ii is poasibj $g calculate tbo molecular weight of the parent
hydrocarbon jhc.
1! ~ 2C
s x SIjj 9 ber V Id the molecular imlgbt of tbs oemponont
barring x ohloriaa atoms. 7ese* for n t 42$ ad 0 Cl we will get
* v
426 > 260 +.&o 156 end equal with the other nolekyle*
r
ffbo soot probable formula with carbon and hydrogen giving this solaoular
weight lo Cjg B^p and ^thie can only be satisfied when the paroat^hydro-
earbon is biphenyl, end the unknown being polychlorinated biphenyls,
2hi a explanation was later fully Verified by Inlection of a synthetic
ysc tax the oass spec,
furthermore* extensive gas ohromatograpblo investigations proved that the
790 standard gave peaks with the case retention tins ae the unknown *
peaks froa the eea eagle*
*
(
tilth tbs aethod loot dsoeribed Z suppose that os bavs a new possibility
to study the-residues in the air because the pins needles can allways be
tie have had groat difficult; in quantifying the 7CBt
but when getting a little mors Mas it will be passible, tie have
done a few calculations on a few* epoiev and X suppose they are right
within u footer 2* tie hare found the residue to bp from
.
It has been sqt statement hare to-day to preeent thin method for studios
of defiling of tbs mature, and with this method * new typo of defiling
agents has been found to be present in nature, and a few experiment Haro
shown where they may be found*
**
.
Bow this method le going to be used in the first hadn to estimate bow the
situation is in nature as a whole, and in the other hand to find the
looks tfcroug which they find its way to nature. 5oa maybe are prooent
here today to get news about the leaks, and to then Z want to oay coco
bwk la a )r..
.
. - mbW7
phgncr 2000095
MONSFOX00000096
WATER PCB-SD0000054856
So xnueb X thialr X safe coy agal that the PCS hardly ess eat fros
asrleulture* Ae support for -this suggestion X oaa say
w have found
PCB Xa eagle feotWro iron SikemuMet frca 1944, sbero botfly *oy
ehlOriantoft pesticides sere used la eerleuitruD On* mm thJLag that X
fifed laporteat to say la that la contrast to the seroory problsa this does
cot eeos to Vo a pure dcedleh problaa. X have lost studied efcro*tegraD
telcos Area Xoadpa air, and they olocxly oontaln Kt, nt tor* Hpldoa h&o
toil bo* that ho also fifed tbm Is Me fiBh&snploe* Sut finally la vaiting at or rotniXts Xsahould Ilk# to polat8Se iwething* Xt is proved that POT
ooaOe to feature so dont taw doiv Aero they ere usod, tut they ere vo*y
persistent to cbosdoale and to fire* X thick the poison J*7 should try
to state that a oostest of PCS shall atloays bo found lo sa open deolaro*
tioa* ,
*
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PHGNCR 200009$ MONSFOXOOOOOQ97
WATER PCB-SD0000054857
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WATER PCB-SD0000054861
PHGNCR 2000101 MONSFOXOOOOOI02
WATER PCB-SD0000054862
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