Document 10Vq2Vqa3e0LyDq57o5qr1v3Z

The Finding of Chlorinated Dibenzofurans in aJapanese Polychlorinated Diphenyl Sample by Johje A. C. Roach and Inwm H. PoMENAnri /NriMOfl oj (,'AmtVa/ T+rhnn!t*$y im liruf Afiminimratim lT'*thmgtnt D.C. 20204 Thn presence of tetra- and poncachlorlnacod dlbncourans and hexa- and hepeachlorinaced naphthalenes in two samploa of polychlorinated biphenyl mixture* (PCB) of European origin has been reported by VOS et al. (1970a). Their Interest in a content enelysle of the ewe PCD sampioe for Impurities was stimulated by previous work in which toxic effects observed In chicks were attributed to thees two samples (VOS st al., 1970b). Kennehlor 400 (KC-400) > e coenerclsl PCD preparation of Japanese origin, hos been identified sa the causative agent of Turtle, a human poisoning inetdsnt resulting from ehs ingestion of dee oil contaminated with KC-400 (KURATSUNB et si.. 1972). Wo hove analysed a sample of KC-4Q0 made available to us from the Kanagafuchl Chemical Industrial Co., Led. through Dr. H. Dtunenthnl of the Food end Drug Administration co determine whether potentially haxardout chemicals, particularly chlorinated dlbenaofwrane (Cl-OBF), were present as contaminants. It was the objective of this analysis to obtain, where possible, structural Infonsation on ehsee contamlnanea and an estimate of their con centration la the KC-400. Experimental A hexene eoluclon of the KC-400 was gas chromatographed, using an electron capture dstactor, on e column of 80-100 sh Chromosorto WRF eoeted with 10% DC-200 and on a column of 157. QF-1 and 101 DC-200 each coated separately on 80-100 tush cbror.iosorb VHP end mixed in a 1:1 weight ratio. The KC-400 (chlorine coneent 48%) shoved only minor diffsrencee in peek locations and relative peak height# from Aroeior 1249 (chlorine content 48%), a PCB of American manufacture. Therefore, Aroclor 1248 eo which a reference mixture ef 41-, trl- and tetraehiorlnatad dibancofurana (prepared by chlorination of dlbanxofucan in eatrachloroethylane with iron powder an catalyst) had been added, vet used by Porter to devise a procedure to separate Cl-DBF from PCB (PORTER, 1971b). This procedure, an used in the present study. Is outlined In Figure 1. STS HONS 087364 it t taut I* f m lated i ehe ml m* ih QF-l >rb mtonc tire of .'trtfwa >a red KC-400 in hexane solution Florltll Column /\ KB In tluaet A (discard) Cl-DBF In iluat* B / Alumina Micro Coliam (PORTSR, 197U) / KB is eiuate 1 (discard) \ Cl-DBF in eiuate 2 Figure 1 Procedure for Separation of Chlorinated Dlbansofurana from Chlorinated Btphenyli in KC-400 The Floriall (8 g of 60-100 math PR grade) va# activated by heating at 130*C ^ 12 hr and cooled to rooa temperature in a deeleeator. Before the eanple va# added to the column, the Florlall coluan vaa prevaahed with about 25 ml hexane. The KC-400/ hexane eoiutlon (.< 2 ml) vae placed on the Floriall, which was contained i# a 10 m i.d. x 300 ma glese column fitted with a Teflon stopcock end medium porosity fritted disk. The column wea eluted with two eluants la succession ec the race of about 3 ml/ min. Eluent A wee 150 ml hexene. Eluent B wee 100 ml 57. ethyl ether in hexene. Eiuate B was concentrated to about 1 ml bafore the sample was placed on the alumina micro column. The micro eolwm vaa a disposable plpet of roughly 5 rm bore Into which a email vad of glees wool had been inserted to retain the aluminum oxide. MerckD alumina, vhieh had been heat-actlvatsd exactly Ilka the FLorisil, vaa poured Into the column to a dspeh of 2.5 cm. Sample or elating solvent ves then added to the eoluan from a disposable plpet or e targe glese syringe. Eluant 1 was 10 ml IX methylene chloride in hexane; eluant 2 was 6 ml 20% methylene chloride in hexane. Additional pcb vaa removed by con centrating eiuate 2 ta about 1 nl and eubjactlng it to a second 339 MQNS 087365 aUmlna micro coluan cleanup, simply repeating the alumina colusm procedure. Ibis procedure wee first applied co duplicate 2 me portions Of KC-400. Electron capture gas chromatography of aluate 2 shoved a gas chromatographic peak in each sample that had a relative retention time matching that of a tetrachlorodibnnsofuran on both a 10X 0V-101 and a 1:1 10% DC-200 + 15% QF-l colusm under the following conditions: glass column, 6 ft. x 4 mm id., column tonparaturo 200*C, injection temperature 225`C; carrier gas, nitrogen at 120 ml/min.; detector, tritium source (eoneencric typo), operated in electron capture mode ee 210*C. Detector voltage was adjusted so that l ng heptachlor epoxide cause* 1/2 full scale recorder deflection et 1 x 10~9 AFS. the reference material for this ratantlon tiaa comparison was the tetrachlorodlbensofuran component in the mixture of chlorin ated dlhensofurans. TP obtain sufficient material from the KC-400 for combined gas ehromatography/maaa spectrometry (GC/MS) analysis, the pro cedure vna scaled up co permit cleanup of 8 mg portions of KC-400. Cleaned up aluatea (aluaca 2) vers then combined to give evo composite samples, each equivalent to approximately 100 mg KC-400. These two eompoeitee were analysed by GC/MS (Finnlgao 1015 quadrupole meet spectrometer) and the resulting data wars examined for evidence of the preeence of compounds chat were not ehlorobiphenyls. A reagent blank of equivalent size was also analyzed by GC/MS. The reagent blank contained none of the compounds found in the samples. The qualitative results of the KC-400 analyses ere summarised In Table 1. The Identification of the compounds listed in Table 1 was baaed on e comparison of the mass spectra of the sample compounds with the toast spectre of the di-, trl-, end tetrachloclnated dlbensofuran components in die reference mixture end on comparison with mass spectral data on chlorinated dibenaofurane in the published literature (VOS et el. 1970a; ratSTCH* et al., 1972; PLBMER ec si., 1973). The amount of taerachlorodlbensofuran contained In the reference mixture was approximated by comparing Its nicrocoulometric GC response with tha response given by e known quantity of a tatrachlorodibensodloxin of known structure# Tha tetrachlorodlbenaodloxln was considered an appropriate mlcrocouloaetrle referonea conpound because of lea structural similarity to tatraeblorodlbensofuran. The electron capture GC responses of tho tetrachloro component in ehe synthetic mixture and In the KC-400 entreats vara than compared to quantitate the tetrachlorodlbeasofttraa la tha KC-400. Tha KC-400 waa aaelaatad to contain approximateLy 1 ppm tetrachlorodlbenso.uraa. 340 MON* 0b7366 COlUMl : tima :: showed 'vt bn both the mn ' Le r 1/2 eon una orln- iLned proi`C-400. 0 KC-400% amined eralytad i found yeee undo vtra at fixture la* ula* uy of loco* I of eho iC*400 Ibeaeo- TABU 1 Coapoimdt Priintt In Kanechlor 400 After Cleanups Mesa (No. of Cl) and fragmentation Oeta Compound Present Molecular Walghe _gf Compound 259 (3Cl);M-2Cl trlchloroblphenyl 256 270(3Cl);M-(CO+Cl) trlchlorodlbensofuran 270 29D(4C1) ;M-2Cl tetrschlorobiphenyl 290 ( 299(5Cl)iM-2Cl pontachloronaphthalana 298 304(4C1) ;M-(C0*C1), M(CO+3Cl) tetrachlorodlbansofuron 304 312 (5C1) ;M-C1 unidentified; perhaps meehylpentachloronaphthalene or pentachlorophenyl cyclopentediene 312 331(4 or SCI); M-(C<H3Cl) indi cated poeeiblo psntaehlorodlbensofuran 339 ^Listed in order of elution from 31 0V-101, 6 ft. x 2 i.d. glees colissn et 220#C, carrier flow 20 ml He/min.; the eema IS evidence wae found in boch samplee examined for all entries but the last (poeaibla pentechlorodibensofuren) for which e second determination was not ateemptad. Chlorinated dlbonsofurana, including 1 ppm of eccrachlorinacod dibeueofuraa(s), and pentachloronaphthalene have boon identified by combined gaa chromatogrephy/mass apectrometry in a polychlori nated biphenyl of Japanese manufacture (KC-400). The question of diethar chlorinated dlbonsofurana in KC-400 may have contributed to the reported Yueho Incident le thue raised* Acknowledgement Ilka authors wish to thank Sandra Mercisr for assistance in the preparation of the samplee for CC/KS enalysle* The reference mixture of di-, tri-, and tetrachlorinated dlbenzofurans wae kindly provided by Dr Albert Pohland, Food and Drug Admlnleeracioa. Reference# fUHSTONI, D., J. R2S3, K. L. BROWN, R. P. BARRON, end J. N. DAMXCOt J* Aaa. Offie* Anal. Cham. 5J, 95 (1972). Ml HONS 087367 KURATSUNB, H., T. YOBHIHURA, J. MATSUZAKA, and A. YAMAGUCHI: Environ. Health Ptrspocc. Iaaua No. 1( 119 (1972). PLIMMER, J. R., J. M. RUTH, and E. A. VOOLSCW: J. Agr. Food Chon, ii, 90 (1973). PORTER, M. and J. A. BUR**: J, Aaa. 0lc. Anal. Chan. 54, 1426 (1971a). PORTER, M.: PrlvaCa Coanwnlcatlon (Fab., 1971b). VOB, J. 0., J. R. HOEMAM, H. L. VAN DBt MAAS, M. C. TER NOEVER DR BMW, and R. H. DE VOS; Fd. Coanat. Toxicol, J, 629 (1970a). vot, J. e. and J. H. KODIAK: Toxicol. Appi. Fhancol. 17, 636 (1970b). fl I Ci * 1 M f P 'S C 5 > 2 ? t1 9 8, SJ! Ml MONS 067360