1 15 5 https://www.nal.usda.gov/exhibits/speccoll/files/original/9e15e5f09773a0aa98ee0140181c7e97.pdf b2224a743dd75784251e31d5f0d189a8 PDF Text Text Item D Number 02323 Author Benfenati, E. Corporate Author Report/Article Title Typescript: Polychlorinated Dibenzo-p-dioxins (PCDD) and Polychlorinated Dibenzofurans (PCDF) in Emissions From and Urban Incinerator. 2, Correlation Between Concentration of Micropollutants and Combustion Conditions Journal/Book Title Year Month/Day Color Number of Images Dflscripton Notes Monday, September 24, 2001 Page 2328 of 2337 �accepted, Sept, '82 POLYCHLORINATED DIBENZO-p-DIOXINS (PCDD) AND POLYCHLORINATED DIBENZOFURANS (PCDF) IN EMISSIONS FROM AN URBAN INCINERATOR. 2. CORRELATION BETWEEN CONCENTRATION OF MICROPOLLUTANTS AND COMBUSTION CONDITIONS E. Benfenati, F. Gizzi, R. Reginato, R. Fanelli, M. Lodi" , R. Tagliaferri° Laboratory of Environmental Pharmacology and Toxicology "Mario Negri" Institute for Pharmacological Research, Via Eritrea 62, 20157 Milan, Italy ° "Ecolab s.r.l." coop., Vignate, Milan, Italy v B E6 I 0 NE L0 M BA RD IA Ufftcso Spaciale di-Ssvoso Via S.Carlo, 4 - 200SG SEViESC /rn INTRODUCTION In a previous paper (1) we. repgrted the average and peak values of PCDD and PCDF emitted from an urban incinerator monitored over a 9-month period. We underlined the wide variability in the different sample concentrations and the need to study the possible correlation between the amount of micropollutants released and several combustion parameters. Three hypotheses are currently supported about the presence of PCDD and PCDF in incinerator emissions.. Lustenhower et al. (2) and Esposito et al. ( ) suggested that these compounds are 3 present in the burned waste and are not destroyed; Rappe et al. (4) considered chlorophenates and other similar compounds as the precursors of chlorinated dioxins and furans. Finally Dow Chemical Company (5) argued that there is a very large series of organic compounds that can be considered as precursors and so chlorinated dioxins and furans occur in all combustion processes. In order to investigate the main parameters which .decomposition of PCDD and PCDF during combustion, temperature, stack flow and tons of refuse burned PCDF, HC1, NOXl S02, CO and particulate were also may be- involved in the formation and/or incinerator operating temperature, fume were recorded and, in addition to PCDD and analyzed. METHODS The method of sampling (effected on two consecutive days each month) and analysis for PCDD and * PCDF have already been described ( ) HC1, S02 and particulate were collected during six hours 1. in isokinetic conditions by a glass probe inside the stack. The analytical method for HC1, adsorbed on 0.1 N NaOH, was -that reported by Zall et al. ( ) SO was analysed according to 6. West and Gaeke (*7), with the addition of sulfamic acid (8) and ethyl enediamine tetraacetic acid ( ) Particulate analysis was gravimetric. 9. �Table 1 - Emissions from the stack and combustion conditions (I PART) Sample 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 PCDD(a) 1064 2618 1474 117180 104308 3019 1593 2026 8951 1563 8581 621 . 846 1130 870 • 2538 1548 5881 PCDF(a) -<«> 8036 5648 .111916 97328 6763 3354 2175 9672 1747 3968 1109 1048 2106 1717 ! 3304 2171 6959 In PCDD 6.970 7.870 7.296 11.671 11.555 8.013 7.373 7.614 9.100 7.354 9.057 6.431 6.741 7.030 6.768 7.839 7,345 8.679 In PCDF Minimal Average combustion combustion temperature(°C) temperature f°c) _(0 8.992 8.639 11.626 11.486 8.819 8.118 7.685 9.177 7.466 8.286 7.011 6.955 7.653 7.448 8.103 7.683 8.848 1000 Jc> _(c) 320 460 1010 . 820 680 740 990 950 960 960 . 940 950 1010 1010 600 1032 _(c) _(c) 627 903 1062 1032 894 902 1041 1016 1012 1030 1010 1027 1039 1038 833 i: pg emitted for one ton of waste combusted Data not available For NO2c six 30-minute samplings were .carried out each day. The NOx was determined with the Saltzman reagent (10); NO was oxidized with KMnO, to N02 and then analyzed as exposed. For CO six spot samples were taken each day and CO was analyzed by GC after methanation ( 1 . 1) Fume temperature and stack flow were recorded several times during each sampling. The combustion chamber operating temperature was recorded by an automatic thermocouple throughout sampling. RESULTS AND DISCUSSION Table 1 shows the results relative to each sampling; the possible correlation between all the analyzed parameters and PCDD or PCDF concentrations were investigated using a multiple regressiot method. The only significant correlation was found between PCDD or PCDF concentrations (logarithmic values) and the minimal combustion temperature reached during sampling (Fig. 1 2 : ,). �Table 1, - II. PART ample Mean stack flow (Nm3/h) 1 2 3 4 5 6 7 • 8 9 10 11 12 13 14 15 16 17 18 28500 35700 35700 35000 33000 40450 39300 39650 35350 31500 31000 32750 34450 34500 . 38800 35000 33900 32100 Mean stack Waste combusted temperature HCl(b) (tons/day) (°C) 90 90 130 30 80 90 90 70 80 90 90 90 90 90 - 90 90 90 60 so2<b> co<b> JO JO _(0 JO 203 196 185 200 204 185 143 159 150 159 173 175 172 181 172 173 150 1093 228 1007 .280 7728 304 2592 361 662 444 1824 1455 4356 1033 4666 630 336 294 248 . 438 778 854 974 686 538 1030 879 1365 908 701 474 958 ™" — 51 28 11477 189 182 121 322 30 < 8.4 < 8.4 < 8.4 < 8.4 < 8.4 < 8.4 < 8.4 < 8.4 TT Wb> Particulate(b) JO 596 285 1434 649 563 607 808 445 664 545 608 698 254 675 374 446 "™ JO 3.2 23.7 264.2 «175.2 713.0 565.0 62.5 107.5 137.8 62.8 250.8 210.4 232.8 336.0 375.1 361.9 ^ (b)g/ton: g emitted for one ton of waste combusted ( )data not available c The correlation between average combustion temperature during sampling and the amounts of micropollutants (logarithmic values) emitted was not significant. This suggests that the lowest temperature reached is the main factor affecting the amount of micropollutants emitted» No significative correlation was found between fume temperature,stack flow, HC1, NOX, S02, CO and micropollutants emitted. The similarity between the amounts of PCDD (ar PCDF) emitted on the two consecutive days every i month (see Fig. 3,4), even.when the other parameters change, stresses the importance of the . materials burned in determining the composition of fumes emitted. In fact, the composition of the waste may wedl be similar on two consecutive days. Combustion temperature and HC1 could play a role even in the formation or destruction of theindividual isomers. However no correlation was found between minimal combustion temperature �15-, r = - 0.8347 p < 0.0001 T F 500 1000 Fig. 1 - x = minimal combustion temperature reached during each sampling (°C). y » PCDD emitted (In jug/ton) 104 r HJ.8386 P< 0.0001 i 500 Fig. 2 t r 1000 x «• minimal combust ion temperature reached during each sampling (°C) y - PCDF emitted (In jug/ton) �r = 0.8368 P<0.01 10- i—r 10 15 Fig, 3 - x =* PCDD' released the first sampling day- of each month (In y * PCDD released the second sampling day of each month (In jug/ton) Samplings 1 and 18 are missing because only one sample was taken in those.7 months. r = 0.8704 p < 0.005 10- T—r 5 i—i—r~ 10 Fig. 4 - x » PCDF released the first sampling day of each month (In jug/ton) y » PCDF released the second sampling day of each month (In |Ug/tpn) Sampling 18 is missing because only one sample was taken in the month.. and the ratios HexaCDD/OctaCDD or Tetra CDF/OctaCDF. HexaCDD and TetraCDF are the most abundant isomeric classes* for PCDD and PCDF, respectively, and OctaCDD and OctaCDF are the least abundant. Similarly it was not possible to find any correlation between the amount of HC1 and the ratios' HexaCDD/OctaCDD and TetraCDF/OctaCDF. Morepver the ratio between' individual classes of isomer.s was not affected by the fume temperature, indicating that selective condensation of individual �classes, does not.occur in the stack at least in these particular operating conditions. Fhe amounts of PCDD and PCDF in the different samplings were very similar (see Fig. 5). This suggests that both PCDD and PCDF concentrations depend on common factors. r = 0.9465 p < 0.00001 10- 10 Fig. 5 - x PCDD emitted during each sampling (In jug/ton) PCDF emitted during each sampling (In jug/ton) The data presented here confirm that the emission of PCDD and PCDF from urban waste incineratorsis inversely related to the combustion temperature; this agrees with observations by other authors, who report that TCDD decomposes at temperatures higher than 800°C (12,13) and that noPCDD can be measured in incinerators operated at high temperatures (< 1200°C) ( 4 . However no? 1) information can be drawn from these results on the mechanism by which the combustion temperature* influences the amount of PCDD and PCDF emitted. A low operating temperature could increase the. probability of formation of polychlorinated compounds, reduce the rate of degradation of these compounds once formed or act in both directions. Further investigations are required to clarify the processes of formation and the destructions of polychlorinated pollutants in urban incinerators, but some practical indications on the possibility of drastically reducing the amount of PCDD and PCDF released into the environment: can already be drawn from the data presented here. i ' . REFERENCES 1. F. Gizzi, R. Reginato, E. Benfenati and R. Fanelli, Chemosphere, 11, 577 ( 9 2 . 18) 2. J.W^A. Lustenhouwer, K. Olie and 0. Hutzinger, Chemosphere, 9_, 501 (1980). 3. M.P. Esposito, T.O. Tiernan and F.E. Dryden," Dioxins", EPA, Cincinnati, Ohio ( 9 0 . 18). �4. ,C. Rappe, S. Marklund, H.R. Buser and H.-P.. Bosshardt, Chemosphere, 7_, 269 (1978). 5. Dow Chemical Co.,"The Trace Chemistries of Fire. A Source of and Routes for the Entry of .Chlorinated Dioxir? into the Environment!1, Midland, Michigan USA (1978). 6. D.M. Zall, D. Fisher and M.Q. Garner, Anal. Chem., 28, 1635 (1956). 7. P.W. West and G.C. Gaeke, Anal. Chem., 28, 1816 (1956). 8. J.B. Pate, B.E. Ammons, G.A. Swanson and J.P. Lodge Jr., Anal. Chem., 37, 942 (1965). 9; N. Zurlo and A.M. Griffini, Med. Lav., _53_, 330 (1962). 10. B.E. Saltzman, Anal. Chem., J26,. 1949 (1954). 11. K. Porter and D.H. Volman, Anal. Chem., 34, 748 (1962). 12. R.H. Stehl, R.R. Papenfuss, R.A. Bredeweg and R.W. Roberts, inl'Chlorodiqxins—Origin and Fate1! Blaire E.H. ed., American Chemical Society, Washington, B.C., p.119 (1973). 13. R.R. Bumb, W.B. Crummett, S.S. Cutie, J.R. Gledhill, R.H. Hummel, R.O. Kagel, L.L. Lamparski, E.'V. Luoma, D.L. Miller, T.J. Nestrick, L.A. Shadoff, R.H. Stehl and J.S. Woods, Science, 210, 385 (1980). 14. G.A. Junk and J.J. Richard, Chemosphere, 10, 1237 (1981). � Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Title A name given to the resource Alvin L. Young Collection on Agent Orange Description An account of the resource <p style="margin-top: -1em; line-height: 1.2em;">The Alvin L. Young Collection on Agent Orange comprises 120 linear feet and spans the late 1800s to 2005; however, the bulk of the coverage is from the 1960s to the 1980s and there are many undated items. The collection was donated to Special Collections of the National Agricultural Library in 1985 by Dr. Alvin L. Young (1942- ). Dr. Young developed the collection as he conducted extensive research on the military defoliant Agent Orange. The collection is in good condition and includes letters, memoranda, books, reports, press releases, journal and newspaper clippings, field logs and notebooks, newsletters, maps, booklets and pamphlets, photographs, memorabilia, and audiotapes of an interview with Dr. Young.</p> <p>For more about this collection, <a href="/exhibits/speccoll/exhibits/show/alvin-l--young-collection-on-a">view the Agent Orange Exhibit.</a></p> Text A resource consisting primarily of words for reading. Examples include books, letters, dissertations, poems, newspapers, articles, archives of mailing lists. Note that facsimiles or images of texts are still of the genre Text. Box The box containing the original item. 093 Folder The folder containing the original item. 2328 Series The series number of the original item. Series IV Subseries IV Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Creator An entity primarily responsible for making the resource Benfenati, E. F. Gizzi R. Reginato R. Fanelli M. Lodi R. Tagliaferri Date A point or period of time associated with an event in the lifecycle of the resource 1982 Title A name given to the resource Typescript: Polychlorinated Dibenzo-p-dioxins (PCDD) and Polychlorinated Dibenzofurans (PCDF) in Emissions From and Urban Incinerator. 2. Correlation Between Concentration of Micropollutants and Combustion Conditions Subject The topic of the resource incineration https://www.nal.usda.gov/exhibits/speccoll/files/original/eb3a0f653bc6c863caddf5bc819dbdbc.pdf d0a6a8d81b69aefc041ddd6b323097b2 PDF Text Text Item D Number 02331 Author Rizzardini, M. Corporate Author Report/ArticleTltlO Typescript: Toxicological Evaluation of Urban Waste Incinerator Emissions Journal/Book Title Year 1982 Month/Day November Color D Number of Images 6 DBSCriptOn NOtBS Chemosphere, submitted November '82 Monday, September 24,2001 Page 2331 of 2337 �Chemosphere Submitted, Novemberr82 TOXICOLOGICAL EVALUATION OF URBAN WASTE INCINERATOR . ' - ., ; EMISSIONS :.M« Rizzardini, M. Romano, F. Tursi, M. Salmona, A. Vecchi, M. Sironi, F. Gizzi, E, Benfenati, S. Garattini and R. Fanelli Istituto di Ricerche Farmacologiche "Marie Negri" Via Eritrea 62, 20157 MILAN, Italy ft.EGIONE L O M B A R ' DI A • . • Ufficso Specials di Sevesp' • Servizio Documentazibns Seie Via S. Carlo, 4 . - , 20030 SEVESO (Ml) INTRODUCTION •'"_". .The- incineration of solid urban waste raises- serious problems related to the presence of highly toxic micropollutants in the emissions of incinerating plants. In particular polychlorinated dibenzo-p-dioxins(PCDD) and polychlorinated dibenzofurans (PCDF) mixtures have been found in these emissions (1-5). Assessment of the toxicological risk associated with these mixtures is a major, still unsolved problem. In fact only a few of the 75 PCDD isomers and 135 PCDr isomers have been studied as regards toxicological properties (6,7), and even scarcer are the data on the toxicological effects of their mixtures, most of them artificially built by joining some individual isomers (8-13). In a first attempt to study the potential toxicological properties of a PCDD/PCDF mixture released from an urban incinerator, we carried out an experiment to characterize its immunodepressive and enzyme-inducing properties. 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDi) has been shown to. be a very potent immunosuppressant in animals (14), single doses of 1 jig/kg depressing antibody production for more than 40 days ( 5 . Little information is 1) available on the immunosuppressive effects of 2,3,7,8-tetrachlorodibenzofuran (TCDF) in laboratory animals ( 6 . .Very recently TCDF was compared with TCDD for its toxicity on the 1) immune system of mice ( 7 . It is generally believed that these two compounds induce 1) similar toxic effects (18) when 10-30 times more TCDF is given. We reported (17) that the humoral response is more sensitive than cell-mediated reactions (Graft versus Host reactionmitogenic responsiveness) to single TCDF exposure and that the immunosuppression induced by TCDF recovers faster than that induced by TCDD, antibody induction being totally restored by day 40. We thus chose to investigate the effects of extracts from an incinerator on humoral antibody production, using heterologous erythrocytes as antigen. An artificial mixture containing known amounts of TCDD and TCDF was prepared as reference control, to evaluate the interaction of the two toxic agents or their possible synergism. TCDD is among the most potent mixed-function oxidase (MFO) inducing agents yet demonstrated in mammalian liver. Low dose studies in the rat gave an ED5Q of TCDD induction .of benzo(oO pyrene-hydroxylase of 0.63 ug/kg and the smallest dose of TCDD which significantly increased enzyme activity was 0.002 ug/kg (19). In mice (C57B1/6J) an £050 of 0.29 ug/kg TCDD for hepatic AHH induction was estimated (20). �4' * TCDF is closely related to TCDD in structure and acute toxicity ( 8 . Its bio1) logical activity (ED5Q for AHH induction) is almost equivalent to TCDD in the chick embryo (21). In mice (C57B1/6J) and ED50 of 7 ug/kg TCDF was found (20) (AHH induction). An important observation was that there is an excellent correlation between AHH induction and the toxic potency of certain PCDD and PCDF isomers ( 2 . Despite the large 2) amount of information about the single compounds none is yet available on the effect of a mixture of TCDD and TCDF on these parameters (P-450 content and hepatic MFO enzyme system). It was therefore deemed interesting to investigate the inducing properties of a mixture of the two compounds and of extracts from an urban waste incinerator, containing complex mixtures of PCDD and PCDF. " - ' - - - . . / - ' .; MATERIALS Animals ' • , - . , • ' " . • • C57B1/6J male mice where obtaine from Charles River, Calco. (Italy) and used when 10 weeks old* Chemicals ' ' . . • All chemicals involved in the extraction, purification and analysis of the PCDD and PCDF micture have been reported ( ) TCDF was a kind gift from Dr. C. Rappa, University 5. of Umea, Sweden. ' • " ' ' • "^ ' EXPERIMENTAL The PCDD and PCDF mixture was drawn from a modern municipal incinerator in Italy, equipped with electrostatic precipitators. Gas condensate collection and the methods of extraction with n-hexane and of purification were as previously described ( ) An LKB 2091 5. GC-MS equipped with an LKB 2130 computer was used for GC-MS analysis, in the reported conditions (5). An imal_t r eatment After analysis, the sample (mixture 1) was dissolved in 10 ml of acetone-corn oil (1:6 v/v) and given as a single i.p. injection to mice (7-8 per group) as such or diluted 10 times (mixture 1/10), at a dose of 10 ml/kg. TCDD (1.2 ug/kg), TCDF (10 ug/kg) and the mixture of TCDD and TCDF (respectively 1.2 ug/kg and 10 ug/kg) were dissolved and given as described for mixture 1. '~~~' groduction assay_ Q 4- 10° sheep red blood cells were injected i.p. 7 days after treatment with the different toxic agents and plaque forming cells (PFC) in the spleen were counted 5 days later by the Yeme's technique (23). �1 statistical analysis -r- _ - Results are presentad as means +_ s.e.; the statistical significance was evaluated by Duncan's test* Enzyme indue tion_ass ay Mice were killed by cervical dislocation and livers were immediately removed, rinsed in saline and blotted dry. The tissues were homogenized with an Ultra-Turrax apparatus in 0.05M phosphate buffer pH 7.4 (1:4 v/v) and centrifuged at 9000xg for 20 min in a refrigerated centrifuge. The supernatant fractions were stored at -70°C until used for cy to chrome P-450, 7-ECD and protein assay. Cytochrome P-450 was determined according to Omura and Sato (24) . 7-Ethoxycoumarin 0-deethylase activity was assayed according to Greenlee and Poland. ( 5 . Proteins were measured according to Lowry et al. (26). 2) ' ' • ' * . - . . , RESULTS The amounts of individual classes of PCDD and PCDF employed for the experiment are listed in Table 1. They correspond approximately to the amount of micropollutants •released in about . . seconds or the amount obtained from about 0.5 kg of waste ( 7 . 05 2) Table 1 - PCDD-PCDF dose administered (ng/kg) , corresponding to mixture 1 PCDD PCDF a Tetra Penta Hexa Hepta Octa a 3688 (17.5) 4816 (22.9) 7152 (34.0) 4332 (20.6) 1064 (5.0) 8800 (35.3) 7696 (30.9) 4540 (18.2) 3004 (12.1) 880 (3.5) Total 21052 . 24920 In brackets the percentage of the individual classes of isomers. Effect_on_antibody_groduction The effect of an artificial mixture of TCDD and TCDF at a ratio of ' about 1:8 was first investigated. As shown in Fig. 1, TCDD inhibited antibody production by 80% while TCDF at the dose used was not depressive. However, when the two toxic agents were administered together in the same solution, the degree of inhibition (50%) was significantly lower than that induced by TCDD alone (80%) when results are expressed either as PFC/10° splenocytes or as PFC/spleen. Thus, the presence of TCDF can modify the immunosuppressive capacity of TCDD. The simultaneous administration of 3.688 ;ug/kg tetrachlorodibenzodioxin and 8.8 jug/kg tetrachlorodibenzofuran (mixture 1) or of a dose 10 times lower (mix I/ 1C) did not significantly modify the immune response, antibody production being only 15% inhibited. �EFFECT OF SIMULTANEOUS ADMINISTRATION OF TCOD AND TCDF AND OF MIXTURES FROM INCINERATOR ON ANTIBODY PRODUCTION. PFC/10 6 splenocytes r? RFC /spleen 100- o cc H~ z o u u. 50O ** TCDD TCDF TCDO TCDF Mix. Mix. 1/10 1 TCOO TCOF TCDD TCOF Mix. Mix. 1/10 1 •FIG. 1 TCDD (1.2 ug/kg), TCDF (10 ug/kg) and extracts from an incinerator were dissolved in acetone:corn oil (1:6 v/v) and given i.p. on day -7. 4-108 SRBC were given i.p. on day 0 and the test was done 5 days later. Mix 1 = extract containing 3.7 and 8.8 ug/kg of tetrachloro-dibenzodioxin and -dibenzofuran respectively; see Table 1. • Mix. 1/10 » dose one-tenth Mix 1 Effect_on_CYtochrome_P-450_content_and_on_7-etox2Coumarin-0-deeth2lase activity Twelve days after a single dose of 1.2 pg/kg TCDD, cytochrome P-450 content was still about 3 times higher than, in control mice. As expected, there was a shift of 2 nm in.-the peak absorption maximum (from 450 nm to 448 nm; see,Table 2). This shift was also observed when TCDD was administered simultaneously with TCDF. No effect on maximum peak absorption was observed with TCDF alone. In this respect, no clear-cut response was seen when incinerator extracts were given. Parallel to cytochrome P-450 induction, a ten-fold increase was observed in the activity of 7-etoxycoumarin-O-deethylase. TCDF at 10 ug/kg had no inducing effect. When the same doses of chlorinated compounds were given in combination, an induction pattern similar to that caused by TCDD alone was observed; however the content of cytochrome P450 was only doubled in comparison to controls and was significantly lower (p^O.05) than in mice treated with TCDD alone. In the same treatment groups the activity of 7-ECD was 8 times higher than in control mice, but somewhat lower (1179 pmol/min/mg prot. versus 1315) than in TCDD treated animals. The effect of an extract from urban incinerator smoke containing 3.3 and 8.8 ,ug/kg of tetrachloro-dibenzodioxins and dibenzofurans was then evaluated: cytochrome P-450 content �», significantly raised and this inductive effect was more evident on 7-ECD activity. — ten times lower dose of incinerator extract had no effect on cytochrome P-450 or 7-ECD, ' Table 2 - Effect of simultaneous administration of TCDD, TCDF and incinerator mixtures on cytochrome P-450 levels and 7-ethoxycoumarin-O-deethylase activity. Treatment Cytochrome P-450 or P-448a (nmol/mg prot) Vehicle 0.63 +:0.028b ( ) 6c 2,3,7,8 TCDD 2,3,7,8 TCDF 1.87 0.75 1.57 0.97 0.82 TCDD +• TCDF Mix 1 Mix 1/10 + 0.07** (6) £0.04- (6) +_ 0.18**+(5) ^ 0.06* (5) -t- 0.017 ' 6 () 7-Ethoxycoumarin-O-deethylase (pmol/min/mg prot) 143 + 6 (5) 1315 +_ 62 (6)" 173 1179 352 234 ^ 15 (6) +_ 98 ( ) 5' +_ 40 ( ) 6' + 16 (6) +•* p^O.Ol from control group (Duncan's test) *- p^O.05 + pjgO.05 from mice treated with TCDD alone (Duncan's test) 3 . Cytochrome P-450 indicates the microsomal co-binding pigment found in control mice, b Mean + S.E. c In. parentheses the number of observations. For treatment schedule refer to Fig. 1 I CONCLUSIONS Our preliminary results indicate that microsomal enzyme induction is a prompt, sensitive biological parameter of exposure to PCDD and PCDF, even when these compounds are in mixtures whose composition is not completely known. The observation that the administration of a dose of TCDF inactive per se simultaneously with an active dose of TCDD reduced the immunosuppfessive and enzyme inducing capacity of TCDD suggest a competitive effect at the receptor level. From this finding with a given TCDD:TCDF ratio, however, it* is impossible to extrapolate the effects of different ratios or of higher active doses of TCDF. These unexpected biological effects illustrate the complexity of problems to be tackled in order to improve the prediction of toxic effects of mixtures of chemicals released simultaneously in the environment. For the time being, statements on the harmlessness to humans of urban waste incinerator emissions appear untimely (28). / ACKNOWLEDGEMENTS The authors express their gratitude to M. Lodi and R. Tagliaferri (ECOLAB s.r.l. Coop., Vignate, Milano, Italy) for sampling and to Dr. C. Rappe (Department of Organic Chemistry, University of Umea, Sweden), for .supplying the TCDF standard. �'•-.. ^1. K..;.01ie, P.L.. Vermeulen and 0..Hutzinger, Chemosphere, £, 455-459 (1977).* t 2. H.R. Buser, H-P. Bosshardt and C. Rappe, Chemosphere, 7_, 165-172 (1978). 3. G.A. Eiceman, R.E. Clement and F..W. Karasek, Anal. Chem., 51, 2343-2350 (1979). f j 4. J.W.A. Lustenhouwer, K. Olie and 0. Hutzinger, Chemosphere, 9, 501-522 (1980). . ' i c ~~ 5. F. Gizzi, R. Reginato, E. Benfenati and R. Fanelli, Chemosphere, 11, 577-583 (1982). [ 6. M.P. Esposito, T.O. Tiernan and F.E. Dryden, "Dioxins" tJ.S. EPA ; Cincinnati, Ohio (1980). 7. J.A. Goldstein. Halogenated Biphenyls, Terphenyls, Naphthalenes, Dibenzodioxins and Related Products, Elsevier, Amsterdam, R.D. Kimbrough (ed.), 151-190 (1980). ; 8. M. Nishizumi, Toxicol. Appl. Pharmacol., 45, 209-212 (1978). —*—* 9. S.. Oishi, M. Morita and H. Fukuda., Toxicol. Appl. Pharmacol., 43, 13-22 (1978). i i ] 10. H. Bauer, K.H. Schulz and U.Spiegelberg,Arch.iGewerbepathol.Gewerbehyg. 1_8_,538-555( 1961). ! 11. 5. Kawano and. K. Hiraga,, Jpn. J. Pharmacol.,, 28, 305-315 (1978). "" ' 12. S^ Oishi and K. Hiraga, Food Cosmet. Toxicol., 16» 47-48 (1978). 13. K.D. Courtney, Bull. Environ. Contain. Toxicol., 16, 674-681 (1976). 14. J.G. Vos, R.E. Faith and M.I. Luster. Halogenated Biphenyls, Terphenyls, Naphthalenes, Dibenzodioxins and Related Products, Elsevier, Amsterdam, R.D. Kimbrough (ed.), 241-266 (1980). 15* A. Vecchi, A. Mantovani, M., Sironi, W. Luini, M. Cairo and S. Garattini, Chem. Biol. Interact., 20, 337-342 (1980). 16. M.I. Luster, R.E. Faith and L.D. Lawson, Drug Chem. Toxicol., 2_, 49-60 (1979). 17. A. Vecchi, M. Sironi, M.A. Canegrati and S. Garattini, Symposium on Chlorinated Dioxins and Dibenzofurans in the Total Environment, Ann Arbor, Science Publishers, L.H. Keith, G. Chondhary and C. Rappe (eds.), 1983, in press. 18. E.E. McConnell and J.A. Moore, Ann. N.Y. Acad. Sci., 320, 138-150 (1979). 19. K.T. Kitchin and J.S. Woods, Toxicol. Appl. Pharmacol., 47, 537-546 (1979). S'. *"" """ 20. A. Poland, E. Glover, M. DeCamp, C.M. Giandomenico and A.S. Kende, Science, 194, 627-630 (1976). 21. A. Poland, E. Glover and A.S. Kende, J. Biol. Chem., 251, 4936-4946 (1976). 22. A. Poland, W.F. Greenlee and A.S. Kende, Ann. N.Y. Acad. Sci., 320, 214-230 (1979). 23. N.K. Jerne and A.A. Nordin, Science, 140, 405 (1963). 24. T. Omura and R. Sato,'J. Biol. Chem., 239, 2370-2378 (1964). 25. W.F. Greenlee and A. Poland, J. Pharmacol. Exp. Ther., 205, 596-605 (1978). 26. O.H. Lowry, N.J. Rosebrough, A.L. Farr and R.J. Randall, J. Biol. Chem., 193, 265-275 (1951). 27. E. Benfenati, F. Gizzi, R. Reginato, R. Fanelli, M. Lodi and R. Tagliaferri, Chemosphere, in press. i / "28. U.S. Environmental Protection Agency, Environmental News, Nov.19 (1981). _, } � Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Title A name given to the resource Alvin L. Young Collection on Agent Orange Description An account of the resource <p style="margin-top: -1em; line-height: 1.2em;">The Alvin L. Young Collection on Agent Orange comprises 120 linear feet and spans the late 1800s to 2005; however, the bulk of the coverage is from the 1960s to the 1980s and there are many undated items. The collection was donated to Special Collections of the National Agricultural Library in 1985 by Dr. Alvin L. Young (1942- ). Dr. Young developed the collection as he conducted extensive research on the military defoliant Agent Orange. The collection is in good condition and includes letters, memoranda, books, reports, press releases, journal and newspaper clippings, field logs and notebooks, newsletters, maps, booklets and pamphlets, photographs, memorabilia, and audiotapes of an interview with Dr. Young.</p> <p>For more about this collection, <a href="/exhibits/speccoll/exhibits/show/alvin-l--young-collection-on-a">view the Agent Orange Exhibit.</a></p> Text A resource consisting primarily of words for reading. Examples include books, letters, dissertations, poems, newspapers, articles, archives of mailing lists. Note that facsimiles or images of texts are still of the genre Text. Box The box containing the original item. 093 Folder The folder containing the original item. 2331 Series The series number of the original item. Series IV Subseries IV Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Creator An entity primarily responsible for making the resource Rizzardini, M. M. Romano F. Tursi M. Salmona A. Vecchi M. Sironi F. Gizzi E. Benfenati S. Garattini R. Fanelli Date A point or period of time associated with an event in the lifecycle of the resource 1982-11-01 Title A name given to the resource Typescript: Toxicological Evaluation of Urban Waste Incinerator Emissions Subject The topic of the resource incineration dioxin animal testing enzyme induction https://www.nal.usda.gov/exhibits/speccoll/files/original/7a6af2afc295e2cf4a85cc403460cc16.pdf b995faea331db8445d2ec040daa6a4d6 PDF Text Text Item D Number °2342 Author Bonaccorsi, A. Corporate Author RdpOrt/ArtlClB TitlB Typescript: Study of the Bioavailability in the Rabbit of the TCDD Present in Powdered Soil from Seveso Zone A (Milan) Journal/Book Title Year 1983 Month/Day June Color Number of Images D 29 Descrlpton Notes Friday, October 05, 2001 Page 2342 of 2422 �LSTISA^r 1383/19 Study of the bioavailability in the rabbit of the TCDD present in •pondered soil from Seveso Zone A (Milan) ' 1 o * o 1 A.Bonaccorsi , A.di Dorrtenico*", R.Fanelli', F.Merli , R.Motta , . R.Vansati 2,f G.Zapponi. 2 1 Istitutq di Ricerche Farmacologiche' "Mario Mecri", Milan 2 Laboratory of Comparative Toxicology and Ecotoxicology, Supreme Health Authority, Rome Study conducted under tlis agreement befvesii the Special Departrier.t of Seveso {'/.ilan) of the Lombard-/ P.egio:-., tl.e "Mario Hegri" Pharmacological Research Institute and the Istitxito Superior® di Sanita (rLoma^ of 8 Jur.e 1321 �1 INTRODUCTION TCDD (2,3,7,8-tetrachlorodibenzo-p-dioxin) is tiie most toxic of the polychlorodibenzodioxins and is, as a rule, an unwanted microcontaminant of 2,4,5-trichlorophenol, produced on a large scale by industry. TCDD has also been formed accidentally in relatively high quantities in a few accidents at chemical plants, including the one at the 1CMS3A chemical plant (Seveso, Milan) in July 1976. This event led to diffuse TCDD contamination of the environment whose gravity is still visible 5 years later (1). Free TCDD has been the subject of numerous toxicological studies, mostly designed to characterise its toxic properties rather than to assess its absorption by the organism (2), However, exposure to the poison in the environment is often due to the presence of environmental substrates to which TCDD is in some way bound; more rarely is the compound fouiid in the free form. Thus the present study arose from the need to establish the bioavailability of TCDD --hen the substance is administered to the rabbit on pondered soil and not -in the free form. 3ir.ce previous work had shown that the liver is the target organ in that species (3), accumulation of TCDD in the liver was taker, as index of absorption. The various forrrulations containing TCDD '"ere administered by gastric tube. The extent of bioavailability may be of importance in ar. assessment of the toxicological risk. 2 METHODS OF ANALYSIS The determination of TCDD in particulate soil was dons by mear.s of a known GC-M5 method (4) adapted for small samples ;'~-2 c] and v.sing t'/.e following sts^s: extraction in a Soxhlet ar^arav.is, �chroma tography on a multilayer column (described later) and chroinatography on a column of activated alumina. The determination of TCDD in liver tissue was done by a GO-MS method described in the literature ( 5 ) , using the following steps: alkaline digestion, extraction with a solvent, chromatograp hy on an Extrelut column and chromatography on a column of activated alumina. As shown in Table 1, recovery by the method as a whole and by two purification steps is on average always more than 9Q% in the case of powdered soil samples. In the case of liver samples recovery calculated with the addition of a tracer is B0%. For further details see Appendix. 3. PREPARATION OF THE FORMULATIONS TO \g of dry agricultural soil was removed from an area in Zone A with a high level of contamination (mean TCDD >3COO ug/rr/) . The stone-free material was spread on plastic sheets for drying in the air (t =s 25-30°C) to constant weight (weight variations <2% over 24 h), crushed vigorously, remixed and sieved for collecting the powder fraction with a particle si-ze of between 200 and ^00 mesh. This fraction (-~0.5 -eg) was further mixed in a rotary evaporator at a moderate speed for about 1C h to obtain a product with a high degree of homogeneity. The results of tan analyses or. as many portions of powder (on average ^2 g each) taken at random revealed a mean contamination of Si - S pro (Table II). The standard deviation value denotes a satisfactorily homogeneous distribution of the contaminant. Portions of tl'.is powder (2.00 0.05 c sach) were administered to rabbits r see Treatment of the rabbits) . �TABLE I Mean recoveries of TCDD for the methods of analysis used and for two specific purification operations. Subject Multilayer column Alumina column Soil 1 2 Soil 23 Soil 34 Hepatic tissLia"3 1 2 3 4 5 number of data 71 22 20 16 Mean recovery (°0 96-12 94- 5 92±15 31 9^i 8 94- 7 57 30-10 For further details and for the individual data on each subject see Table "Recoveries of TCDD" (Appendix). Laboratory-contamined soil analysed \vithin £3 h of addition of TCDD. Laboratory-contaminated soil analysed after "aging" ( 1 month 37 Yield of TCDD (Cl ) added to uncoiltaminated ?articulate soil and to Seveso1 particulate soil before extraction, in the range 10—1 CO ppb. •3 7 • / Yield of TCDD (C1J/) added to the sample of hepatic tissv.e before alkaline .digestion, in the ranee C.5-5 ppb. �TABLE II TCDD levels determined in ten samples of powdered soil from Zone A after sieving and homogenisation. „,__,„ ,. _ Recovery2 Sample ., g 1 2 3 4 5 6 7 8 9 10 2.15 1.80 2.06 2.11 1.92 2.32 2.00 1.89 2.00 2.00 TCDD found *g 174 96 131 94 160 173 141 122 100 91 93.0 103.5 99.5 95.7 80.0 90.0 110.0 107.5 94.5 101.5 203 159 180 173 157 186 159 146 139 137 ppb 94.4 88.3 87.4 82.0 81,8 80.2 79.5 77.3 69.5 68.5 Mean TCDD3 concentration and its standard deviation: 8 1 - 8 ppb 1 Number of refluxings for the extraction in a Soxhlet apparatus. „„ 2 Calculated on the quantity of tracer (I?.CDD(C1J/)) added to the soil before extraction. 3 '.To correction "/as made for losses due to the method of analysis because of the high recoveries (p>97%') . �10 kg of soil presumably free from TCDD but morphologically similar to that o£ Zone A was treated as described in the preceding section to get-^1 leg of ponder (200-^00 mesh). Analysis of five samples («* 2 g each) revealed no TCDD at an analytical sensitivity of 0.6 ppb. This powdered soil was administered to rabbits as such in the tolerance tests (see Treatment of the rabbits). One croup of accurately weighed portions (1.00-0,02 g each) of the same powder was spiked with TCDD in an acetone solution (<3 ml/portion) at a level of 10.0-0.2 or 40-1 ppb); the portions were left to "age" in the dark at 2C°C for over 1 month before administration to the rabbits. A second group of portions of pov/dered soil was contaminated (40-1 ppb) 48 h before administration to the rabbits, sufficient time for the solvent to evaporate. Each portion (or the combination of two) "/as the single daily dose intended for each animal. Some samples of each group were used for testing the recovery in the critical steps of the analysis method. The following were prepared: two solutions of TCDD in an acetonevegetable oil mixture (1:6) containing 1C.C-G.2 and 40.0-0.3 nc/ml of poison respectively, and a'water-alcohol solution (l:l) containing 40.0-0.3 ng/ml. The single daily doses intended for each animal were 1 .00-0.01 and 2.0C-0.01 ml according to case. 4 T2EATKSITT OF TKS ?.A33ITS Before treatment of the animals started, tests were done to find out whether the repeated administration of riven quantities of powdered soil could cause digestive tract disturbances that micht affect the absorption of TCDD from the gut.. The tolerar.ee tests �were done on six rabbits, each of which received a suspension of powdered soil (2.0 g) in water (10 ml) by gastric tube daily for 7 days. The animals presented no ill-effects, apart from a brief interruption of weight gain on the first day of treatment. 3y the second day the bodyweight growth curve had already resumed its normal trend. The previously described preparations were dosed by gastric tube for 7 consecutive days to groups of 5-7 male white rabbits of mean starting weight 2.4-0.2 kg (Fig. 1), caged singly. In all, 12 groups of animals were treated with the following preparations (daily doses in brackets): a) powdered soil from Zone A, Soil S (^80—160 ng daily); b) "aged" laboratory-contaminated soil, Soil 2 (20-40-80 ng daily); c) laboratory-contaminated powder, Soil 1 (40-80 ng daily); d) acetone-oil solution (20-40 ng daily); wateralcohol solution (40-80 ng daily). The daily dose (1.00 or 2.00) of powdered soil was sxispended in 10 ml of water. The preparations containing TCDD were given regularly at 09.00. On day 3 the animals were killed and the livers removed and prepared for analysis. Table III gives (a) the weights of the animals at death (p = 2.5 -0.3 kg; see also Fig. 2), (b) the weights of the whole livers (see also Fig. 3) and (c) of the portions of hepatic tissue used for analysis, -(d) the individual TCDD levels found in the hepatic : tissue: in the case of groups or. broadly homogeneous treatments (as "/ill be noted frcm the lack of significant differences betweer. the specific values of the means) the cumulative mean was calculated on several homogeneous groups. Table III contains three such cases: treatments with TCDD in solution at the dose of 40 r.c daily, treatments with laboratory-cor.taminsEi powdered soil at the dose of 40 ng daily -and treatments with laboratory-contaminated powdered soil at the dose of SO nc daily. The results are surjur.arisec in Table IV �Table III Levels of TCDD found in the hepatic tissue of rabbits after seven days treatment Treatment BodyLiver weight.. (e_) weight 1 Total Sample 2 (kg? 1.90 2.43 2.72 2.55 2.57 2.65 2.57 2.57 2.73 2.70 2.70 2.32 73.9 70.0 107 84.1 73.0 90.1 98.0 86.9 98.0 100 118 80.0 22.0 21.0 22.0 22.0 20.0 19.0 20.0 18.0 18.0 20.0 21.0 20.0 1.85 2.20 Oil-acetone 2.35 2.33 2.47 2.35 2.25 40 ngj/die ALcoholj-watei 2.35 2.31 2.30 2.57 2.05 2.67 40 ng/die 2.80 Acohol-watei 2.75 2.95 72.8 82.9 79.2 61.5 66.9 96.5 92.4 96.9 81.4 77.7 100 62.9 72.0 88.0 86.5 108 18.9 22.0 23.3 23.6 24.8 23.5 22.5 23.6 23.3 22.8 25.7 21.0 18.1 17.0 21.7 20.2 20 ng/die Oil-acetone 20 ng/die Soil 2* 40 ng/die 40 ng/die Soil 1 s 40 ng/die Soil 2 2.57 2.40 2.30 2.30 2.51 2.99 2.69 2.35 2.65 2,71 2.20 2.70 2.35 TCDD Found 3 TCDD found ( n g ) ( p p b ) mean value Cppb ) 7.78 6.04 5.83 5.19 3.26 6.58 6.46 5.14 5.13 4.93 4.24 2.52 29.8 29.2 27.7 27.6 26.7 •18.0 29.3 25.6 .23.7 22.2 22.4 13.1 29.3 23.9 27.1 19.9 0.353 0.288 0.265 0.236 0.163 0.347 0.323 0.286 0.285 0.246 0.202 0.126 1.57 1.32 1.19 1.17 1.08 0.766 1.30 1.08 1.02 0.972 0.870 0.626 1.62 1.41 1.25 0.984 0.26 ± 0.07 0.26 ± 0.08 i lfl igi8 4> n Qft u ys •f 0 77 ' . /-\ -^ w 2. ( ] • £ // W / i \J » M M , „, ± 0.27 1<J1 Mean value for the three groups 1.1 ± 0.5 45.6 68.5 1.60 28.5 31.7 41.9 1.32 79.2 16.7 71.4 21.2 0.786 n QQ j_ Pi . *J b O T f. 20.4 91.2 .762~\ w « yo ± U J 26.8 O 28.2 19.8 0.701 93.0 113 56.4 25.6 0.701 27.6 24.3 80.0 0.880 0.807 19.3 81.4 23.9 0.652 19.4 12.6 73.6 16.1 0.586 0.66 ± 0.15 110 27.5 87.7 12.6 0.572 22.0 74.4 15.4 0.560 27.6 0.559 23.6 13.2 63.3 �Continuation of Table: "Levels of TCDD in the liver") Mean value for the two .groups 0.81 ± 0.51 80 ng/die 2.80 Alcohol-water 2.40 2.85 2.90 2.80 80 ng/die 2.80 2.93 Soil 1 2.53 2.85 3.15 80 ng/die 2.60 2.85 Soil 2 14.4 17.2 12.6 16,2 20.1 16.0 18.0 18.5 24.0 21.8 18.7 21.0 21.0 17.0 19,0 • 2.72 2.55 ^80 ng/die s.oj.1 3s 2.65 2.90 2.84 2.75 3.18 2.90 2.70 Mean value for the last two groups 1.5 ± 0.2 21.6 27.9 85.0 1.29 21.8 103 26.6 1.22 19.3 111 18.2 0.942 0.814 0.88 ± 0.28 115 20.5 16.7 19.7 128 13.6 0.692 105 17.8 0.6"3 12.0 21.7 11.7 116 0.540 M.60 ng/die 1.92 2.50 T f O 2.58 2.22 2.43 2.75 2.27 75.0 80.1 A A r\ 94.9 88.0 90.2 100 89. S 2.65 Soil S 20.0 20.0 20.0 22.0 21.0 21.0 25.0 1/"S /™S 49.1 35.4 32.6 38.8 45.2 28.8 32.2 30.7 28.2 25.4 33.9 33.4 32.3 26.1 25.7 3.41 5.11 2.59 2.40 2.15 1.80 1.79 1.66 1.17 1.16 1.81 1.59 1.54 1.53 1.35 85.0 83.1 94.9 119 114 93.0 113 109 111 125 97.2 80.0 100 103 98.0 77.2 58.0 41.6 45.5 40.5 31.2 23.0 i i r 5.86 2.90 2.08 2.07 1.93 1.48 0.922 1 2.73 ± 0.52 1.S2 ± 0.52 1.56 ± 0.16 * /"\O 2.18 ± 0.96 1 At the end of treatment 2 Portion of fresh liver used for the determination 3 Values corrected for losses inherent in the method of analysis used (last column of Table on "Recoveries") ^Laboratory-contaminated soil left to "age" ( 1 month) before administration 5 Laboratory-contaminated soil administered with 48 h of spiking with TCDD 5 Powdered soil from Seveso (mean contamination: 81 ppb) �TA3LS IV Mean TCDD levels in the liver of rabbits treated for seven- consecutive days Tith various formulations containing the poison. Vehicle TCDD ng/day 4 Solvent Soil 2 Solvent Soil 1-27 o Solvent Soil 1-2 Soil S'2 Soil S 20 20 40 40 80 80 • 80 'I ww Number of specimens TCDD 'opb) in the liver x - 6? T t QC ^ 0/ ^ - >, P y /«* / 5 7 16 13 0.26-0.07 0.26-0.08 0.12-0.40 0.15-0.37 1 .1 -0.3 0.81^0.31 0.94-1 .3 0.54-1 .1 5 10 7 2.7 ±0.5 1 .5 -0.2 0.33^0.28 1 .7 -3.8 1 .3 -1 .8 0.43-1 .3 7 2.2 -1 .0 0.34-3.5 * 1 For further details and for data on the individual experiments see Table "Levels of TCDD found in hepatic tissue". 2 Mean values and standard deviations. 3 Confidence interval 4 Oil-acetone 5 Laboratory-contaminated soil left to "age" (> 1 month) before administration. 6 Combination of the results for treatments -"ith alcohol-v/ater solution and those of oil-acetone solution. 7 Combination of the results for treatments \7ith the tv/o formulations of soil prepared: Soil 1 and Soil 2, the first of vliici '•/as dosed v/ithin 48 h of spilcing '"ith TCDD. 8 Alcohol-v/ater. S' ?ov/dered soil from Seveso (mean contamination: 31 o^b) . �CONCLUSIONS Table V summarises the results of the statistical analysis (ANOVA and Duncan's test) conducted on the data given in Tables III and IV. Regarding the bioavailability of TCDD, our conclusions are as follows: (a) at the lowest dose tested (20 ng daily) no statistically significant difference in bioavailability emerged between TCDD in solution and TCDD on laboratory-contaminated powder; (b) at the doses of 40 and 80 ng daily the bioavailability of TCDD distributed over powdered soil appears on average to be 29-and 44% respectively, lower than that of TCDD in solution; the lower limits of the confidence intervals (99/0 give a decrease in bioavailability of 5% and 19% respectively; (c) The bioavailability of•TCDD present on the powder of soil taken from Seveso, contaminated after the 1CMESA accident, .is on average 68% less than that of TCDD administered in solution; here again, however, the lower limit of the corresponding confidence interval (99/0 supplies a much smaller variation in bioavailability (-40%); (d) statistical analysis of the results obtained with treatments at 40 ng daily shows that there are no significant difference: in TCDD absorption between recently contaminated oowdered sci: (Soil 1) and TCDD in solution, in two cases out of three (see Table III); (e) there is a significant difference (-45% on average) between the bioavailability of TCDD present in the soil from Sevesc and that of TCDD on laboratcry-contami-^sd soil; ir. this case the lower limit of the confidence interval '9S?0 supplies 2.;-. estimated decrease in bioavailability of only 5%; �10 TABLE V Statistical analysis of tlie results obtained (previous table) for tlie evaluation of variations in bioavailability. TCDD Formulations (ng:day) compared ^ 20 40 80 80 80 1 2 3 4 5 5 7 3 Soil Soil Soil Soil Soil Significance level of the variation Mean reduction (%) of bioavailability x CI (S9%)3 2/Solvent nonsignificant 1-2/Solvent5 p <.01 6 1-2/Solvent p < .01 S/Solvent7 p < .01 29 44 68 S/Soil 1-28 43 p < .01 5.0 - 53 19 - 68 40 95 5 - 81 • AN OVA and Duncan test In each pair the TCDD of the second formulation (Solvent) was assigned maximum bioavailability (100%). This was arbitrarily attributed also to the TCDD of the second formulation of the Pair Soil S/Soil 1-2. Confidence interval Laboratory-contaminated soil after "aging" versus solution in .oil-acetone. Laboratory-contaminated soil ("aged" and "not aged") versus solution (in alcohol-v/ater and oil-acetone). Laborator2/>-contamined soil ("aged" and not "aged") versus solution in alcohol-water. Povdered soil from Seveso versus solution in aicohol-v/ater. Powdered soil from Seveso versus laboratory-contaminated soil ("aged" and not "aged"). �11 (f) two highly significant linear regressions may be derived, from the data obtained, vis. from the treatments with TCDD in solution and with TCDD venicled by the laboratory-contaminated powder (Fig. 4); the two regression lines permit an estimate of the mean levels that might have been present in the liver following treatment with the above formulations at the. dose of 160 ng daily: in contrast to what happens in the case of the laboratory-contaminated soil, the value extrapolated for treatments with TCDD in solution is significantly higher than the value determined experimentally for the group of animals treated with powdered soil from Seveso, Soil S, at the: same daily dose (V160 ng daily) . In sum, the data obtained indicate that in the rabbit the absorption of TCDD from orally dosed preparations is clearly less when the poison is in the soil than in. solution. This phenomenon be/ come-s apparent at the highest doses. Acknowledcmen t /' The authors thank the Lombardy Region, Special Department of Seveso, for financing this survey. References (1) Pocchiari, F., di Domenico, A., Silano, V . , e Zapponi, G. (1981) Environmental Impact of the Accidental Release of Tetrachlorodibenzo-p-dioxin (TCDD) at Seveso (Italy). Presentato al Workshop on Human Health Aspects to Accidental Chemical Exposure of Dioxin - Strategy for Environmental Reclamation and Community Protection, 4-7 Ottobre 1981, Bethesda (Maryland, USA). i'2; Poiger, H., e Schlatter, Ch. (1980): Influence of Solvents and Adsorbents on Dermal and Intestinal Absorption of TCDD. Food and Cosmetic Toxicology 18, 477-481. �(3) Panelli, R., Bertoni, M.P., Castelli, M.G., Chiabrando, C., telli, G.P., Noseda, A., Garattini, S., Binaghi, C., Marazza, V., e Pezza, F. (1980): 2,3,~,8-Tetrachlorodibenzo-p-dioxin Toxic Effects and Tissue Levels in Animals from the Contaminated Area of Seveso, Italy. Archives of Environmental Contamination and Toxicology £, 569-7/T: (4) di Domenico, A., Silano, V., Viviano, G., e Zapponi, G. (1980): Accidental Release of 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) at Seveso, Italy: I. Sensitivity and Specificity of .Analytical Procedures Adopted for TCDD Assay. Ecotoxico1ogy and Environmental Safety 4, 283-297. (5) Panelli, R., Bertoni, M.P., Bonfanti, M., Castelli, M.G., Chiabrando, C., Martelli, G.P., Nod, M.A., Noseda, A., e Sbarra, C. (1980): Routine Analysis of 2,3,7,8-Tetrachlorodibenzo-p-dioxin in Biological Samples from the Contaminated Area of Seveso, Italy. Bulletin of Environmental Contamination and Toxicology 24, 818-SUI (6) Baser, H.R., e Rappe, C. (1978): Identification of Substitution Patterns in Polychlorinated Dibenzo-p-dioxins (PCDDs) by Mass Spectrometry. Chemosphere 7, 199-211. �13 APPENDIX Methods of analysis (a) For the oowdered soils: The procedure of analysis for the samples of powdered soil was one that has already been v/idely used for TCDD determinations in various environmental substrates at Seveso (4), adapted for the type of samples under study ar reported later on. For a detailed discussion of the procedure see References. .T The three steps used (extraction, purification on a multilayer column and purification on an alumina column) were subjected to numerous preliminary tests in order to ensure practically quantitative recoveries and a high degree of reproducibility. riT, Extraction, All the samples of powdered soil were extracted in a Soxhlet apparatus for ~24 h (refluxing 4-6 times/ft.) with dichloromethane (--75 rail, HPLC grade) as solvent. The'sample was prepared for extraction by mixing intimately ^2 g of dry powder with 8 g of anhydrous sodium sulfate. The extract "-as left to evaporate slowly under a stream of nitrogen in order to obtain a moist residue. Par if i cat ion on multilayer column. This column (l.D. 1C mr.: total length 250 mm) consisted (from bottom to 'top): glass wool; anhydrous sodium sulfate 0.5 cm; activated silica gel 1.5 cm; mixture of anhydrous sodium sulfate and sodium bicarbonate (4:1 w/w) 2 cm; celite 545 1 cm; mixture of concentrated sulfuric acid and celite 5^-5 (1:1 w/w) 5 cm; anhydrous sodium sulfate 1 cm. The secuence of the lasers is shown in Fig. 1 . Before use, the multilayer column (one for each attraction) was washed -with three portions (i c ral each) of dichloromethane, followed by ~liree portions (10 ml) of :i-sen-cans The residue of extraction was rale en \i^ with ± rn' o- ---^c.--^-- = �14 (HPLC grade) and transferred to tlie top of the column before the oentane washings had eluted cornsletelv; the operation -;as J* *•» J. -• * repeated another seven tines in the same "/ay to ensure quantitative transfer. Percolation 'vas by gravity. The eluate collected (^32 ml) \vas allowed to evaporate to -dryr.ess spontaneous ly. Purification on column or ac'civatgdalumina. Alumina March (neutral aluminium oxide 90, 170-230 mesh) vas Icept in an over, at l30°ci5°C for 14 h and then stored in a desiccator on silica gel ready for subsequent use. The columns (l.D. 10 mm, total length/-*250 mm) -;;ere prepared by inserting first a piece of glass ";ool, fcllov/ed by 5 cm of alumina (r-4 g) and 0.5 cm of anhydrous sodium sulfate. The residue from the multilayer column '^as taken up quantitatively \vith five successive portions (2 ml each) of n-hexane and transferred in succession on to the top of the column. There followed three washings (10 ml each) "/ith a mixture of n-hexane and dichloronvethane (1/0 and four elutions ~'ith a total of 20 ml (5 ml per elution) of a mixture of r.-hexar. and dichloromethane 20%. This eluate (-20 ml) ~;as carefully collected in a bealcer and left to evaporate to a moist residua; this was then transferred quantitatively tn a suitable container evaporated to dryr.ess and diluted to a kncr/n volume for t2:e assa by means of irGC/irMS according to the procedure describee in tl: reference cited. (b) For the samples of hepatic tissue: The determinations on the liver samples ••/•ere cone by means of" a procedure already used for the determination of TCDD in rabbit • liver (5), as reported later. For a ds-ailec discussion of rlie procedure see references. �15 Alkaline digestion. About 20 g of liver <vas hydrolysed "/itl: 10 ml NaOK 10 N plus 20 ml ethanol at 90°C and refluxed- for 1 h. Extraction. After cooling, the sample in the alkaline medium was extracted twice with n-hexane (20 ml each time) . Purification on column of Sxtrelut. A column of Sxtrelut was pretreated with 20 ml concentrated sulfuric acid and left to equilibrate for 4-14 h. The total hexane solution ( ~ 0 ml) .4 '•was left to percolate through the column, which was subs ecu sr.tl; washed with 20 ml n-hexane. The eluate was evaporated to dryness in a stream of nitrogen at room temperature. Purification on column of activated alumina. The columns of neutral alumina (45 x 5 mm), with a layer of anhydrous sodium sulfate at the top (5 mm), were washed with 3 ml dichloromethar.; and activated at 270°C for 12 h or at 4GQ°C 'for 4 h. The residue from the -column of Sxtrelut was taken up with n-hsxane (total volume-*3 nil) and transferred to the top of the column. This was followed by an elution with 5 ml carbon tetrachloride followed by 4 ml dichloromethane. The latter fraction, containing TCDD, was evaporated to dryness. The residue was taken up -*itl dioxane (0.100 ml) and analysed by GC and mass fracmentograp'.v-. * N In some cases samples from croups on like treatment "/ere pooled and analysed by high resolution GC (25000 theoretical plates) aiid MS•according to a method reported in the literature (6). All the reacents and standards used in this study were of a hic'.i degree of purity and were grade H?LC or R3, where possible. Tl'.e TCDD used vas of the same purity as the environmental standards . Table VI and Figs. 5-11 summarise all the results and informaticappropriate for defining the quality of the analysis methods used. �18 Table VI Rcoveries ("£) of TCDD for the methods of analysis used and for two specific purification operations. Multilayer column 141. 0" 123. 5 120. 0 115. 5 114. 6 114. 5 114. 3 110. 5 110. 5 109. 0 108. 5 108.0 107. 5 105. 0 104, 5 104.0 104. 0 103. 8 103. 5 103. 1 103. 0 102. 6 102. 0 101. 5 101. 5 100. 9 100. 8 99. 9 99. S 99. 0 98. 1 98. 0 98. 0 97. 5 96. 0 96. 0 95. 0 95.0 95.0 94.5 94.0 94.0 93.5 91.5 91.0 91.0 90.0 90.0 89.0 89.0 88.9 88.0 88.0 88.0 87.0 86.5 85.5 84.0 84.0 84.0 84.0 84.0 83.5 83.5 82.4 82.0 81.5 81.0 78.0 76.0 73.8 65.1 + 57. 0' Alumina column Soil 1 Soil 2 102.0 101.5 101.0 100.0 99.4 135. 8" 116. 7 109. 4 104. 6 102. 0 102. 0 99. 1 •99. 0 98. 0 97. 6 97. 6 95. 5 95. 0 94. 0 92. 0 87. 5 85. 5 69. 4 66. 6 66. 5 60. 3 106. 0 102. 6 98. 0 97. 5 97. 0 95. 5 94. 0 93. 5 93. 0 92. 5 92. 0 91.0 86. 0 81, 9 81. 4 77. 9 98.0 96.5 95.6 95.0 94.0 93.5 93.5 92.5 92.0 91.8 91.5 91.0 90.0 89.0 87.5 82.7 82.7 74.0" 47.0" 2 1 ': Soil 3' 110.0 107.5 105.5 103.5 101.5 99.5 99.5 99.5 99.0 98.0 97.5 97.5 97.0 95.7 94.5 94.5 • 93.0 93.0 92.5 90.0 90.0 88.5 88.5 88.0 88.0 87.5 86.5 86.0 86.0 85.0 80.0 Hepatic tissue 108. 5" 99. 0 98. 2 96. 6 95. 2 94. 8 91. 0 89. 3 89. 0 88. 6 88. 3 88.0 88. 0 88. 0 88. 0 87. 0 86. 2 86. 0 86. 0 85. 0 85. 0 85. 0 84. 0 84. 0 84. 0 83. 3 83. 3 82. 4 82. 0 80.0 80.0 78.5 78.2 77.5 77.3 77.0 77.0 77.0 76.0 76.0 75.5 75.2 75.0 73.0 72.8 72.2 71.4 71.0 70.2 67.0 65.5 64.3 63.0 63.0 62.0 62.0 61.0 60.4 Means and standard deviations and number of data used in the calculation: -. 94 ± 5 80 ± 10 92 ± 15 92 ± 8 96 ± 12 94 ± 7 {20} {16} {31} {71} {5 7} {22} 1 Laboratory-contaminated soil Laboratory-contaminated soil left to "age" 3 Recoveries of TCDD (Cl ") added to samples of powdered soil and of hepatic tissue before extraction "Aberrant (according to Chauvenet) 2 3 �19 14 12 N Media Dev. St Min Max 3 4 . 1.80 2.22 2.64 3.06 Fig. 1. Distribution by weight group of 44 male white rabbits before treatment �20 28 N 24 Medi a Dev. St. Win Max 7Q 2.6 0.3 I .85 3.18 20 12 8 1 .75 2.25 2.75 3.25 Fig. 2. Distribution by weight group of 70 male white rabbits on day 8 (sacrifice) of treatment. Note (see Fig.l) that the animals presented an average increase in weight in the week of treatment; in some cases there was a slight weight loss unrelated to any specific factor. �20 N Media Dev. St. Min Max = = = = = 70 92 16 61.5 128.0 12 8 56 80 104 128 grammi Fig. 5. Distribution by weight group of the livers removed. from the rabbits after treatment with various formulations containing TCDD. �3.5 . 3.0 y = 0.041x - 0.51 y = O.OZlx - 0.076 2.5 Q. Q. O 2.0 4-) cn 0) 4- 1.5 O O O 1— 1.0 n s 80 120 160 ng/die Fig. 4. Regression lines obtained by using the set of data for treatments with TCDD in solution (Q) and the set of data for treatments with laboratory-contaminated powdered soil ( 0 ) « The significance level of the regressions is F ^ „ = 172.35, P< .001; F \ 8 = 106.67, P< .001 respectively. To highlight the results of treatments with Seveso soil, two points have been plotted ( u) corresponding to the mean values of the TCDD levels determined analytically in the two groups treated. The standard deviation is shown above and below the value of each mean. �23 anidro 1 em 5 em Celite -t- H2S04 (1:1 p/p) t em Calita 2 cm NaHC03 +• Na2SC 1 . 5 em Gel di Silice 1 cm Ma.SO, anidro (1:4) Cotone \ Fig. 5. Multilayer column for the first stage of purification of the extracts. Description in the text. �Z4 28 r 24 L Media Dev. St. Min Max 20 I 16 U 12 h 8 L 4 L 60 Fig. 6. 84 108 recovery 132 Block distribution of percentage recoveries obtained from the multilayer columns (see Table VI first column) �25 12 N Me<i1a 10 = 22 = 94 Dev. St. = 5 M1n * 82.7 Max « 102.0 o 76 Fig. 7. Block distribution of percentage recoveries obtained from the alumina columns (see Table VI, second column) �2b 8 - 6 - 4 N - Media Dev. St. Min Max 2 . 30 64 98 132 recovery .8* Block distribution of percentage recoveries obtained on applying the method of analysis to the determination of TCDD added to uncontaminated powdered soil, within 48 h of addition (Soil 1: see Table VI, third column). �6 . 4 . 2 . 70 Fig. 9. 80 90 100 recovery no Block distribution of percentage recoveries obtained on applying the method of analysis to the determination of TCDD added to uncontaminated powdered soil left to "age" (Soil 2: see Table VI, four-th column) �N 8 - Media Dev. St. Min Max 6 - 4 - 2 . 70 114.8 Fig. 10. Block distribution of percentage recoveries obtained on applying the method of analysis to the determination of labeled TCDD added to powdered soil of different types (laboratory-contaminated or contaminated following the ICMESA accident) before extraction (see Table VI, column five) �29 14 N 12 Media Dev. St. Min Max 10 8 45 57 69 81 93 105 % recovery Fig. 11. Block distribution of percentage recoveries obtained on applying the method of analysis to the determination of labeled TCDD added to hepatic tissue before alkaline digestion (see Table VI, sixth column) � Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Title A name given to the resource Alvin L. Young Collection on Agent Orange Description An account of the resource <p style="margin-top: -1em; line-height: 1.2em;">The Alvin L. Young Collection on Agent Orange comprises 120 linear feet and spans the late 1800s to 2005; however, the bulk of the coverage is from the 1960s to the 1980s and there are many undated items. The collection was donated to Special Collections of the National Agricultural Library in 1985 by Dr. Alvin L. Young (1942- ). Dr. Young developed the collection as he conducted extensive research on the military defoliant Agent Orange. The collection is in good condition and includes letters, memoranda, books, reports, press releases, journal and newspaper clippings, field logs and notebooks, newsletters, maps, booklets and pamphlets, photographs, memorabilia, and audiotapes of an interview with Dr. Young.</p> <p>For more about this collection, <a href="/exhibits/speccoll/exhibits/show/alvin-l--young-collection-on-a">view the Agent Orange Exhibit.</a></p> Text A resource consisting primarily of words for reading. Examples include books, letters, dissertations, poems, newspapers, articles, archives of mailing lists. Note that facsimiles or images of texts are still of the genre Text. Box The box containing the original item. 094 Folder The folder containing the original item. 2342 Series The series number of the original item. Series IV Subseries IV Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Creator An entity primarily responsible for making the resource Bonaccorsi, A. A. di Domenico R. Fanelli F. Merli R. Motta R. Vanzati G. Zapponi Date A point or period of time associated with an event in the lifecycle of the resource 1983-06-01 Title A name given to the resource Typescript: Study of the Bioavailability in the Rabbit of the TCDD Present in Powdered Soil from Seveso Zone A (Milan) Subject The topic of the resource dioxin animal testing soil testing Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Title A name given to the resource Alvin L. Young Collection on Agent Orange Description An account of the resource <p style="margin-top: -1em; line-height: 1.2em;">The Alvin L. Young Collection on Agent Orange comprises 120 linear feet and spans the late 1800s to 2005; however, the bulk of the coverage is from the 1960s to the 1980s and there are many undated items. The collection was donated to Special Collections of the National Agricultural Library in 1985 by Dr. Alvin L. Young (1942- ). Dr. Young developed the collection as he conducted extensive research on the military defoliant Agent Orange. The collection is in good condition and includes letters, memoranda, books, reports, press releases, journal and newspaper clippings, field logs and notebooks, newsletters, maps, booklets and pamphlets, photographs, memorabilia, and audiotapes of an interview with Dr. Young.</p> <p>For more about this collection, <a href="/exhibits/speccoll/exhibits/show/alvin-l--young-collection-on-a">view the Agent Orange Exhibit.</a></p> Text A resource consisting primarily of words for reading. Examples include books, letters, dissertations, poems, newspapers, articles, archives of mailing lists. Note that facsimiles or images of texts are still of the genre Text. Box The box containing the original item. 098 Folder The folder containing the original item. 2555 Series The series number of the original item. Series V Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Creator An entity primarily responsible for making the resource Frigerio, A. R. Fanelli Source A related resource from which the described resource is derived Cronache di Chimica Date A point or period of time associated with an event in the lifecycle of the resource 1978-03-01 Title A name given to the resource Identification and Quantitative Determination of 2,3,7,8-Tetrachlorodibenzo-para-Dioxin (TCDD) in Animals from Contaminated Areas: the Seveso Case Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Title A name given to the resource Alvin L. Young Collection on Agent Orange Description An account of the resource <p style="margin-top: -1em; line-height: 1.2em;">The Alvin L. Young Collection on Agent Orange comprises 120 linear feet and spans the late 1800s to 2005; however, the bulk of the coverage is from the 1960s to the 1980s and there are many undated items. The collection was donated to Special Collections of the National Agricultural Library in 1985 by Dr. Alvin L. Young (1942- ). Dr. Young developed the collection as he conducted extensive research on the military defoliant Agent Orange. The collection is in good condition and includes letters, memoranda, books, reports, press releases, journal and newspaper clippings, field logs and notebooks, newsletters, maps, booklets and pamphlets, photographs, memorabilia, and audiotapes of an interview with Dr. Young.</p> <p>For more about this collection, <a href="/exhibits/speccoll/exhibits/show/alvin-l--young-collection-on-a">view the Agent Orange Exhibit.</a></p> Text A resource consisting primarily of words for reading. Examples include books, letters, dissertations, poems, newspapers, articles, archives of mailing lists. Note that facsimiles or images of texts are still of the genre Text. Box The box containing the original item. 104 Folder The folder containing the original item. 2823 Series The series number of the original item. Series V Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Creator An entity primarily responsible for making the resource Rizzardini, M. M. Romano F. Tursi M. Salmona A. Vecchi M. Sironi F. Gizzi E. Benfenati S. Garattini R. Fanelli Source A related resource from which the described resource is derived Chemospere Date A point or period of time associated with an event in the lifecycle of the resource 1983 Title A name given to the resource Toxicological Evaluation of Urban Waste Incinerator Emissions