Document 2qoRGJN6kLvBoNBgarm3zveL6
/ 1 1 26
NAT
VOL. 221. MARCH 22. 1969
trimmer. K., and Rdrhiimnii. M. K.. Fr,i l`mr..gt. SOS <tOS). KwWfkv. O. \V.f HurttM, K. M., nml Kolwr. II. A., /fit<*\rmilry, 3, 020
(1004).
'* TomlhifuMi. II. V., mid Toner, (5. M.. tlinrhrtni*lry,t. 007 (1003).
11 Vliilglrv, J. K. M,
/MttffAn*, c4r<.l3. 210 0000).
' KaJHIBimlnry. 1'. 1... fl"J.
243. 500 (1008).
'* JUxwftl. I. II. Wllinnrr, K., nml Trncr, <. M.. Biochritiitfry, 7. 202D (4*W*>.
"Olllmn. I.. TUew. TV. Warrington. H. (I. ii , Uirrflfii*i*trn, 7. 3450 < 1068).
von Tlorrmircm. M.. nml Teu^r
" BUIinn. D. H, L., Pace, X. It., aimI Spfrge1mn, 3.. l*>oc. Vs .\'nt A<-nd
.*>*.. 47. i:oano7).
'
"Welch. H. L.. Aatcrindl*, 0. T.. ami Glllmm, p. T.. .SrirUrt, 1Q. u;,o (1008).
* 8hnplr, L.. Kcunw: 473(1008).
FcninnilfM. T., and Angus!. J, T.. ,Yiittnf. tJO
I> Wnchler, tt.. and Flirt, W. J.. .Y-rMfiA ttl. 233 (1000).
" Wrlmiwnn. IVI.v, (i., sior. H.. nnd l'ulM. U.. Ivor. r.s \nf Arm!
67. 1870 (1067).
'
'
H Dc Wnrhtcr, K., Vrrlmucl, J.-JV, nnd Flora, W.. FK/IS />//.. 1. m
Chlorinated Biphenyls in Fish, Mussels and Birds frwi the River Rhine and the Netherlands Coastal Area
by
J. H. KOEMAN
Inicltute of Veterinary Pharmacology and Toxicology, Utrecht. The Netherlands
M. C. TEN NOEVER DE BRAUW R. H. DE VOS
Central Institute for Nutrition and Food Research. Zeist, The Netherlands
Polychlorinated biphenyls are widely used in industry, and they are now making an appearance In the world's wildlife. Besides
being a hazard In their own right, they interfere with the detection of organochlorlne Insecticides such as DDT.
At tile end of 1900 a note in (he .Yctr Scientist1 mentioned A proup of compounds which appear ns unknown peaks in gns chromatograms of the tissues of fish and wildlife. In Sweden it was found that these peaks correspond to polychlorinated biphenyls (Pf'Bsi.
I'CBs are used in industry as lubricants, heat trnnsfer media ond insulators. They are compatible with paints, synthetic resins, varnishes and waxes, nnd improve such properties of these products as their chemical resistance, water r< sistance, r.dhe.'ivem'ss and flexibility5,1.
In 1907, J'CBs uere reported prrsent in fish nnd wildlife in Cirent Britain nnd the Netherlands*-*. In these studies the identification was primarily based on the comparison of tho gas-liquid chromatography retention times in the sample extracts with those in solutions of technical PCM mixtures supplied by the industry. No information was given about the identity of the individual peaks.
Wo descnlie here tlie occurrence and distribution of
PCBs in the River Rhine and the coastal men in the Netherlands. The identification was enniod out lw gachromatography nnd mass spectrometry. For this purpu-e n OH 4 mass spectrometer (Vniian Mat (ImbHi ascoupled to a Vnrinu Aeroemph Oven fv|>e .4 .Vm./J. Particular attention was paid to the design of the intn tin-,between tile two instrument,! in order to obtain I he be-i operating conditions'. 'The relative retention tins p,] were cat mini id with a 2fi4-l/f' Yulian Aerograph L'schroinntogniph with electron capture detection. )'\ iv\ glass columns (6 feet x 1/8 inch), filled with I" per eem DC 200 on Uuschrom Q (80-1 On inesli), were used m luali gas chromatographs, and the columns were operated ni 200 C with helium or nitrogen ns carrier gas.
Tho tissues of tlie indicator organisms used throiigliuut this study acre extracted with petroleum ether m a Soxhlct extraction apparatus after drying with nnlndmo. sodium sulphate. The samples were cl.....ted up bv tit-
TabIt 1. MAKti XTXOERS aXD MMBEM Of CHLnsiXE atoms,'t>Lr.crir (in parenthesf*) of PEAKS WITH RELATIVE RETENTION TtMI> (RP.LATlvr. In PlELHtlX !) LONGER THAN 020 AND hllOllTEIt THAN 100
Sampling
Typo itl uinp!o
0-2f
032
0*33 0 34 0 40
r. (>4H
(157
Oft) 0 05 0-71
0-72
>:l
ii >i;
1 lUinrh (f.rnritnt* tu/ijn*) (u(nl IhmIv lirtl
uxirnt-iH UH17, IUn
3S(0| 250(3) 2j0<3>
3M(3) 2(H)(4) 2U0 (4) 200 (4)
2IHKI)
ll^lf.'t) :i2i;,
200 (4)
1 OphiIhHIhB MltJtiu (Jtt/iin) ImIaI IhhI.v
+
+
rt cutmcM* miOh
Mnrl
*t}nli*) 1CMVK
+
i 8dnl prl (y| nnwitit/lru lanr*>uia(if)
+
loltl IkkIy fxlfjii la
3 SiMvlA If'h (t'fM (Slrrnn TWf/pi'rtri*) 1HH
ilMur ii tit 11-Kg fM nifl n 1 00j, 1000 2P2(0) '
3 Ehirr (Snmat*ria nttJlittimn) liuuo Hrn
i*nO rgg
)000, 1007
4B2 (6)
Thmoriilnr J)\* fl Arnthlnr 1200 Clophfn A 00
+ ++
++
,
4- +
4-
+ + + 4* + +
4. *
2&6(3) 200(4) 200(4) THmlrln 406(8)
256 (3)
Telndrln 400 (8)
200(4)
32s t.M
370 (*) 200 (4)
+ 376 (8)
321 (M
324 <51 324(5)
324 (5)
' 321(5) 324 <M 324(51
With Ur rx rptlnn (if lift!, t -Inilrln xml rnmpniin-l 374 f HI all c tmpnilmla Id.'null -.1 ore r.infer, mli If tr> PCtls. s , At llr cl veil rrlrntlnn I Itn.- :i
l iirrnr*nl In Mu' f*lir<tni:if nurnni. X 11) piT ernt t>C 200 <m < :.i*'lirf<iu Q (*0- 1 fHl mr)t) rninimt *a tm-il. f - -`00 iCnrMrr
N. ( * AO ml mini, \(.i - %
numlirr r.ilniliiliniM nr-* 1.ih.-.1 mm ri ;j;,. Jhr annulling plnrct ;>ro ln*iiv;iI<*! in Kin. '2. JK M - lli xm liU rttlu nynir: ifl*nlrin - 1 .:t.4..'*,n.7,r<.*-ri i.u lil' f'*
1,3f3;i,4,7,7ii hrxithj ilf'f4.7*np lh.Mti iMiln'nXMttirnn.
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NATURE. VOL 221. MARCH 22. I960
1127
inctli)lformnmide partition* and column chromatography over activated flumisil. (Dliimii chromatography was |>erformod in two steps. I''irxt tho apolar compounds including the PCBs wen- eluted with hexane and then a 10 per cent diethyl ether in hnxtuio solution wan used to elute dieldrin and cmJriti. Only tlio results of the first fraction analyses are reported here.
In Tables 1 and 2 the relativo retention `tifnes (relutivo to dictdriu equals I), the mass numbers and the numbers of chlorine utoms/iuolcculo are given. Mass Mix'ctra were taken from all peaks present in the total ion current chromatogram obtained with tho double ion source of the C'H i mass spectrometer'. In Kig. 1A and IS gas chromatograms are given of nn eider extract and a tech nical PCU inixturo (pliPtioehlor DP tj). There is a Btrong resemblance between the elect run capture chromatograms mid the ion current chromatograms.
Until from tile mass numbers and the numbers of ehlnrine atnms/molecule we coneludo that most of the compounds present in the extracts can be identified as It'Us. The retention times increase in proportion to the iueresse in the munls'f of chlorine atoms/moieculc. A XUltuiikuMe I'esemblance-wasdbuud in tho chromatograms orphen-jc-lilor Dl* ft, clnpht'ii A 00 and arochlor 1260. -5 ' IChher chlorinated eumpeunds found tire the fungicide hexachlmnlx'iizeno (HCB), /J.//-DDE and the insecticide teiixlriti. HCB occurs in alt places where indicator organ ism* wen* collected, and bus also l>ecn detected in seed rating and predutory bird* in the Netherlands*. It does iih( mm.mii likely that the widespread distribution in the aquatic environment is caused merely by the small scale urn* of the compound in seed dressing practice. On the other hnmi, it must hr rii.^uM.-i/.cil that 1ICB is a very inert compound which shows n strong tendency to accumu lation in unimal li-sues. Of the DOT derived type of compound* only DDK wus detected, although one PCB intri ft'ii'd with its determination. In a former study DDT and ODD were detected in seals from tho Wadden Sen10. Tcludrin was only found in sea bird*, and from other studies wc can conclude that this insecticide must have originated from a pesticide producing plant near the mouth of the river*10. Both in the roach and the birds a compound with T/sO-71, mass nr 370 and 8 Cl was delected. Further studies concerning tho identity of this
compound niv in progress. Tetrnchlornlrcnzenc and pentn* chlm <>Ih'h/.ciic wen* detected in the roach by the mass sped roiuet or at refill ive ret mi ion t itncR shorter t imn 0'2o.
TIhmv i* u marked roMiMohliUKto Im'Iwihmi the three hii 111111 it i ix m n 'ii* in tlx* identify mid pnflern of the PCBs
it f j ifl/iiie<* mfoniioit imiis longer than 0-71. It semis likely iherefure I hut mionI of (lie p( `B material present m tile Iil 1 mid mmi Idl'd* liv ing m tlie Wndden Sen nrigitiiilc<i Mi tin* River Rhine. The eoitNtal water chiefly flows northwards. It lias bi*en observed that dumpings of chemical* arv diluted to a low degree in the littoral current.
Min
------
-tiq. 1. Uu clifuiHAtosnniii'vr A. an efdrr extract. D, K trclmfral f('|
mixture (phcuoehlor DP ti) add C, quail livtr and brain t.\ trail (tor 4^
^
^.^^^xn^oation,*ee tl text). * "
"
In 1965 a clandestine dumping of copper sulphate was
displaced over a range of 100 kin in a northern direction
within a fortnight. It was diluted by u factor of five11.
Ah can be hccu in the tables, tho lower chlorinated
PCBs occur moro frequently in the tissues of ttic much
than in the sea birds. This may indicate that the lower
chlorinated PCBs arc less persistent, and this has been
I'OhfirmetT try an
AVllh Jnpunugr qtmil vlmrh
Wfcre led n oiet continuing 2,Uoo p.p.m. ol phonoehlor
DP 6 of which the chromatogram is given in Fig. ID. Al^
birds (n20) died between 6 and 55 days after closing
started. A chromatogram representing tho residue in the
tissues (liver and brain) of these birds is given in Fig. 1C.
It can lx; seen that a number of tho compounds present
in tlie original mixture were metabolized, particularly the
lower chlorinated ones.
Residues in. tho bruins and livers of tin* quail were
measured ofi a xemiqunnt il at ive Kettle by peek Iiruiht
eotnpariHon of peak r, =s I 45 using the plienochlur 1)1* 6
mixture an a standard. Residue levels calculated in this
way were about twenty times higher in the quad tissues
than in the livers and brains of the eiders. The quail
Trthlr 2. HASS NCXHER* ASP NC5IBERS OF CHLORINE AT0MS/M0LF.CTLE (IX PARENTHESES) OF PEAKS* WITH RELATIVE RETENTION TIXES (n.WTIV TO MELURIN-l) OF PEAKS LONGER THAN 100
HatiipliiiR art'll
Tt pc f R.riMf'U*
r.
101
M7
1-23
1*45
1-57
1 -00
1-90
2 27
2-67
3-20
3 58
l ItosHl <fsuri<vitn rttfil"*) 1967, 1008
ImkIv fXlroCls DOE 316 (4) 358 (6) 35* (6) 358 (6) 502 <71 358 <0> 392 i T ji 302 7) 302 (7) 302 {i} 42* (8) 4- 302 (7)
I Ormiuilllna (Gubin piiW") total IkhIv fxlrncU S24+(5> + 4- 4- + + + + + + +
106m o Mumim I (Mf/fihig rtl"1i") UM1*
4- + -f + 4- 4- + + + * +
n Kami ri-W.I /<.//<-.
lOfifl
+ 4- 4- + + + + + -f + +
X SanOwIrh tern (Stmm
tiasue ami DDK
'RU cxtrurU 1063. I'.iiUt
316 (4)
35S (6) 358 (6)
358 (0) 302 (7]\ 302 (7) 302 (7) 302 (7) 420 (8)
-f +
3^4 (J>)
324 (31
Klftrr (Sumafrria rxtract* 1086. 1007
Vluruiclilor 1>|* 0 Ai-M'hit/r I'jan nopH/'n A I>
1 ami rgR DDK 310 (4)
3 353(0)
353(6) 3H2(7 ) 302 (7) 302(7) 802 (?) 420 (8)
4- +
324(5) 33S (6)
3T.R (0)
35S (6)
.VS (6) s:.k (d)
(0)
324 (5)
3,`iS (0) 3.*.K un
35s (0)
3.'X (0) 358 (li)
35K (8)
302 <7)
3H2 (7) 3U2(7)
358 D
35K (d) 3*s <0)
302 (7 )
302 (7 ) 302 (7 )
302 (7) 302(7)
(t)
:i02 (7) 302 (7) 302 (7)
302 (7) 302(7) 30J <T)
420 18)
4 20 0*1 *20 (M)
W/fli thert.-fjiNtMi<' UIM' .ill com[w>miuUhlrntihrrinre eonruriujUlf let l*( 'Ha.1 + ..\l Ihr rI vimi rrlvnilun tlim- a prak 1" pr mMil In UirrhnnmlUigtiin, not*** mtili'i T.Hilr 1.
o n-
* nO rsj o
m Q
STLCOPCB4010631
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Nru/
vul wi.-march 22. 1969
The Waddeii Sea nreiM published separately)'
ui1 thin #ltidy
Tho peculiar physical properties and rliemicnl staluliiv
of PCHs mako it likely that the nppliriitioti will in<-n-n>
and that their release into the environment will cimtmut'.
Although fatal concentrations are probably not developed
yet along the Dutch coast, wo suggest that monitoring of
the environment should be carried out regularly iu order
to keep an eye on the further trends in the lovrl of con
tamination. Apart from being a potential hazard in the
natural environment, these PL'Hh also i-epn-sciit an im
portant sourco of interference in the detection of inserti-
cides such as DDE, o./?-DDT. p.p'-DDT and ])DD.
Wo thank Mr J. H. Peniimgs. Miss J. Hnrvink mid
Mr J. M. Eyndhuvcn for their technical assist unco.
Autopsy of tho tpiail was carried out by Mr (I. Vos.
The technical PCM mixtures were supplied l.y I'rudclcc
in France (phenochlor), Monsanto in the UN (urodiloi)
and Bayer in Germany (clophen). Fish fnnn the Jtlune
were supplied by the Department of Sport fishing nub
Professional Inland Fishing in Utrecht.
Bccehetl Dcctmbcr 24, 1068.
developed hydropcricnrd* with a dose of 2,000 p.p.m. Flick et al.'1 also discovered hydropcrieards in White Leghorn cockerels dosed with 400 p.p.m. in their diet. No hydrojiericnrfls were observed in birds which died m
llon*anto Company Ttrh. ftnll, .V. PL-30*. 50 (I960).
Entyelopidin of Chtmiral Terhnolooit (Hit. hv Eller Kirk, R., nnd (Hlimn.
D. T\), tcoond rev. H.. fi, 2H9 <1064).
` HnlniCft. I>. C.. Simmon*. J. H.. nnd Tattmi. .1. 0*0., .Y/Umf. 218.227 OMT).
4 Hoklrn, A. V., and Mitfwlvn, K., yttlurr, 216. 1274 (1007).
XofMiicn, J. II.. Owkrmtp, A. A. O.. Vt^-n. J., Hrnmi-r, K.. II<*>.tli. .1.. /.wart, 1'., v.u. MriN<k, K.. nnd van (loptlrron, If., Mrilnf fiiUtim nti, ,t lMvdhomrtr*Untfhnp(n-H firut, it. *41 U007).
T t*n .Vinner tin llrnnw, M. 4*., nd llnitim-r, (1007).
X. A mil. t'Am, .228. T4I
Knulx-rt MnrmdiT. >t. Ktmu. 1!., U**W\. V. \V . Roburn. J.. nntl Thniutun. J.. Ana/yal, BO. I*1h (Uhmi.
I'.. W.. "
Vo*. J. O.. lirw'man, lluHy A., nnd Bcn*<hri|i. II., Mnlrd. \ Landb*iuuirfttn*chnj>iifn (ftat, 33 (In tin* juvax).
nit, ,/
Koomnn. J. If., nnd vnn nrmlm-n, JI.../. .</>/;/,
3 (Stippl.). liniiii.
11 Knenuin, J. H., Venn. J.. ]lfmiw-r, K,, Hui*m*n, dc f(rou<`f, f... nn<l Koulrn, 1. L., flelgol. W\9rn*rh, Mtrre*nrttri., 17, 075 ODGn).
11 Kllck, It. K., O'Dell. H. 0., rwd t'Mlus, V, A., Poultry (1065).
44 uryi
Spontaneous Mutation
The following two articles are concerned with the genetic control
of mutation rate. The first describes the anti-mutagenic effect of some DNA polymerase mutations on phage T4 mutation rates, and discusses the Implications they hold for the mechanism of action of DNA polymerase. The second compares spontaneous mutation rates In different species, and considers the biological factors which may determine these rates.
Genetic Control of Mutation Rates in Bacteriophage T4
Hkhit.aulk increases in tho rates of spontaneous mutation r usunliy attributed to "mutator genoR*1, tho mechanisms of action of which remain olweura. Mutations in the DNA polymuruM* gene. in bacteriophage '1*4 have, however, been observed to exhibit jM>\verfu| nmt/itor effcctM1,1, suggesting that Hponlun*<jUH mutation niton depend oil tho properties nf DNA polymerases, and also tbnt these polymerases play an important part in btuio selection during DNA Mynthosis.
Wo have therefore embarked on a detailed examination of the effects of DNA polymerase mutations on T4 muta tion rates, to throw light on the genetic control of imita tion rates and possibly on the mechanisms of spontaneous mutation and of information transfer in DNA synthesis as well. This article Hiauinari7.es studies <m temperature.
sensitive (to) mutants of gene 43 (the DNA polymetalgene) obtained from the Cultech collection nn a gilt ! Professor K. S. Edgar. A very large fraction of the-.*mutants turns out to exhibit mutator activity. A furl fin surprising result, has been the discovery of several unitmutator alleles of germ 43 whn h sharply reduce rmilaimu rntes at. many sites in the T4 ^'imiiK'.
The In milfanfH have Iwen loealr/ad uilliin p'lir l.'t
by complemenlalion and recombination. A preliminary map hus been prc'pnn'd from twmfnrtor emsses:. Imi its accuracy is limited by the very strong murker e|l < w frequently observed with alleles of gene 43 (ref. 3 nnd unpublished results of J. W. D. nnd E. T. A.). I In fort y*ono mutants occupy approximately twenty-one sites, nnd most of thorn show litile or no dominance, as is to be expected for a catalytic function. Our test-' fur intracistronie (intcmllelic) coinplementation Imvo been uniformly negative or of doubtful sign tfien nee. These te*t
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