Document zd9v6wL46Vy5qNyEeobkGODgz

STUDY OF THJ'1 POSSIBILITY OF LEAD MOTHYLATIQH ' IN RIVER, BEDS Proposal To International Lead Zinc Research Organization, Inc. .292 Madison Avenue New York, Hew York 10017 From Ethyl Corporation Ferndale, Michigan ^8220 1 September 8, 3.9TO I. iN'j'RODtfCTION In recent years, it has been claimed by; government health authorities and some, other investigators that mercury discharged as an:industrial waste '4 into rivers ultimately transforms into a form which is metabolized by organisms in the river,.ultimately ending up as detectable quantities in. fish. This disclosure appeared very surprising in view of the supposed inertness of mercury! ill: water and its extremely low solubility. The explanation was furnished recently by Wood, et al1,2 who showed that mercury can be '.converted to methyl mercury and dimethyl mercury as a result of enzymatic reactions of the following type which are favored in the environment of river sludges: CH3 : : ^Co^++ IN Methylc obalamin e (red) ++ + Hg AT?Factor III Enzyme \ I/ Co ++ ,1\ / Bis-r (brown) 3 In; view of the reactions which mercury undergoes, it.is logical' to ask whether industrial lead wastes dumped into rivers can undergo similar reactions. Lead is most commonly discharged into rivers as,the plumbic (+2) ion., . If it remained in this form it would be readily available for metabolism by organisms at all levels. However, it has been shown (Alleii et als) that ferric hydroxide arid high molecular weight organic species complex with and scavenge the lead from solution. They believe the lead is then lost quickly to the river or lake bottoms. It is therefore relevant to.ask whether it can become available once more for ingestion by various organisms through a methylation reaction of the type which has been observed with,mercury. If this methylation reaction did occur, it could have important implications to the lead content of all river species including edible fish. It is therefore of importance to determine whether such reactions can occur with lead. 1. Wood, , Kennedy, F.S. , and Rosen, G.G. "Synthesis of Methyl-Mercury Compounds by Extracts of Methanogenic Bacterium" Mature 270 173-37 (1905) 2. Allen, 31.E. , Matson, W.R. , arid Mancy 573-583 (1970). J of Water Roll... Cont. Fed. -1EPI 000295 II.___RESEARCH PLAT] It is proposed to study the possible methylation of lead /using river sludges obtained from -the Mississippi River in the Vicinity of the Ethyl Corporation plant.at Baton Rouge, When this .sludge.is; recovered,/.it will be carefully examined to determine whether: any, organic- lead is present. Such material could conceivably be present as a result of lead being emitted from the plant.'in that form. If there is any detectable organic lead it will be extracted before- any further studies are made vising the-, sludge. We trill then follow the procedure of Wood1 to see whether methylation of lead can occur under conditions where the methylation of mercury is known to occur. For convincing proof of this point in the'case lead does not methylate, we will repeat the work of Wood1 to show that mercury does methylate under these conditions. It may be possible that the sludge as such will be difficult to work with due to factors not known to ,us at present. If this is the case, we would extract the methanogenic bacteria from the sludge. The methanogenic orgalnism was isolated recently by Bryant3. This organism can be grown routinely on hydrogen and carbon dioxide. Cell extracts can be.prepared and the activity of the mixture assayed by methane release (Wood et al4), Methylcobalamine is an excellent substrate for the formation of methane in the presence of the methanogenic bacteria. Thus to conduct the experiment in the absence of sludge, we would follow the procedure of Wood1, using a suitable concentration of soluble lead, ATP (adenosine triphosphate) the methanogenic bacteria and methylcobalamine in the reaction mixture. Following the exposure of lead to the methanogenic environment, the mixture would be deproteinated. If organolead compounds are present, they could then be extracted using an organic solvent. Identification of the compounds could then be made using thin layer chromatography. This method of identification has been used for organonercury compounds, and we anticipate no problems with lead. 3. Bryant, M,P. , Wolin, E.A. , Wolin, M. J. , and. Wolfe, R. S Mi c11roni o] or;ic 59 20 (1967). Arch. fur . Wool, J.!t. , Allam, A.M. , Bril] , U J., and ,W< d fc, r;. n, ?M) OfKo). -2- J. Biol, divv-i.. EPI 000296 To complete this study, we would also examine the possible-formation' , of alkyl leal compounds in the absence of bacteria. It is known that in a reducing, environment. meiMury can be methylated in the presence, of methylcobalamine. A;parallel study will be made with lead to determine whether this reaction proceeds. / : To round out the picture, it is proposed to examine the lea.d content of several species of fish. 3''ish from the upper Mississippi would be : compared with those in the area near Baton Rouge. These data would be examined fortrends .relating to locations in the river, and also; in relation to the lead contents of other foods available in the American diet, based oh much information already accumulated from Ethyl Corporation and other sources.' ' HI. EXPBRIEIICE Ethyl Corporation's Research Laboratories in Detroit have been intensively studying the problem of lead in the environment for many years. Detailed studies have .been made of the processes governing the incorporation of environmental lead into many food crops, and, these results have been very effective in showing that lead naturally present in soils is a major contributor to lead in crops, while only in very few cases Ms. there any measurable contribution of atmospheric lead to the edible portion of crops. As part of its study of lead in foods. Ethyl has set up a number of greenhouses, some of which are equipped with very efficient filters so crops can be grown in comparative tests in the normal atmosphere end in the presence of carefully filtered air. In order to support these studies, Ethyl has built up a major capability in analysis, specializing in the examination of lead and lead compounds in a variety of substrates. In addition to .the very extensive studies on crops and other vegetation, we.have dealt in. depth with lead in such diverse materia as water, milk, fish, and seafood. Our analytical studies on milk have shown that the commonly accepted values for lead in milk are far higher than those actually'occurring 'in practice, due to widespread analytical errors occurring in many; laboratories working in this a-rea. Our equipiv.-n't for performing these analyses covers a wide variety, from vet..chemical'methods through the most, advanced instrumental tcch.nlcues, \lv rd:;u h:avai 1 able, on a contract basis, access 'to el octron mi croprcbe. equipment I'm the Identification of extremely small, quantities of lend come, a;.-- . EPI 000297 IV. STAFF . , The proposed work would he under the immediate technical supervision "'. ' 4 of Dr. Gary Ter IIaar9 Research Associate at Ethyl Corporation's Ferndale 1 .Laboratories..:-. r'For the past six years Dr. Ter Ilaar has been' in, charge of Ethyl's studies of lead in, the environment as described above. He would be-available for the proposed work to the extent of time needed to .'directly supdryise the experimental work described, above. . It is expected ; that this would require one nan month of his time. Dr . Ter Ilaar' s biography is present: in the Appendix to this proposal. , The major portion of. the experimental work would be the responsibility of a fully trained technician reporting to Dr. Ter Haar. Selection of... this technician would be,made from among several individuals with substantial experience in the careful handling and study of materials containing trace quantities of lead. - Analytical work would be the responsibility,of Ethyl's Analytical Section under the direction of Dr. James B. Hinkamp. ..Experts on Dr. Hinkamp's staff would, undertake responsibilljiies as 'appropriate' 'to- their own.specialized, experience in wet chemical. Instrumental, and microamalytical techniques. V.. FACILITIES Ethyl Corporation's facilities at Ferndale are well known to the ILZRO group, and no detailed description is required for this proposal.^ Suffice it to say, that we already have on hand the facilities; and equipment needed to do this work and that .no additional equipment of . any substantial magnitude would be required to carry out the work as described above. EPI 000298 VI. COST Based on the: preceding project outline, we envision that this work can be.accomplished within , a six month period, for a. total" cost' of $25 ,791 . . .Cost: breakdown is as follows: LABOR- Project Personnel ( 6 man months.) Supervision (l man month) Analytical Supnort (2 man months) $ 919^ OVERHEAD (computed at 152^of labor) Total labor + overhead 13975 233.69 SUPPLIES & TRAVEL ` FEE (computed at 7/o of labor + overhead) GrandTotal 1000 1622 < $25791 EP1 000299 APPENDIX , DR. GAR!. TER HAAR, Research Associate, earned his B. S. (cum laude) in- \ chemistry -from'ilppes^CdliegG'vinr;19.5'8- and'his Ph, D.> in inorganic chemistry from ^he University of Michigan in 1962. His doctoral .dissertation was oh a study of tetraboranecarbonyl and some of its derivatives.. In addition, . he taught laboratory classes.in high-vacuum techniques and radioactive tracer studies and spent one summer doing research work using radioactive techniques.. - After joining Ethyl in 19o2, Dr. Ter Haar conducted research in the .field, of metalbrganic chemistry, preparing volatile compounds of uranium and thorium under partial sponsorship of the Atomic Energy Commission. . He synthesized a number.of these compounds, which are highly reactive and unstable, and: demonstrated a high degree of technical ability in their handling and preparation. . . ; Dr. Ter Haar .subsequently devoted l-l/2 years to research on high- energy fuels. He then spent another l-l/2 years as Ethyl's representative at the Argonne National Laboratory in a cooperative program on the determination of traces, of lead and lead-210 in air, water, soil, , and various biological specimens. Since .returning to Ethyl's Detroit laboratories. Dr. Ter Haar has been concerned with the total area of environmental lead and its relationship to man. . In conducting this-work.. Dr. Ter Haar has been responsible for the design and. experiments for the accumulation of data and for its analysis. He has worked closely with the personnel of Analytical.. Research and Services group in developing the techniques used to determine the trace levels of lead present in many of the samples. - The results of some of Dr. Tea- Haar's work at the Argonne National Laboratories and at Ethyl's Detroit Research laboratories were recently presented in. two papers.- at- the American' Chemical Society "Symposium EPI 000300 on Air Conservation" 'held., .at the April, 1969 spring meeting in Minneapolis. Bibliography of Dr. G. Ter Haar "DimethyDamiclodifluorophosphine Tetraborane-8-F2PI](CH3)2B4Hs -- A Stable Adduct of B4H0," with M. : A. ; Fleming and R. W. Parry, J., Am, Chem. Soc. 1767 (1962). "A Study of Tetraboranecarbonyl and Some of Its Derivatives," Dissertation Abstracts 23, 3627-28(1963). "Amine Addition Compounds of H3BCO and BaIIq CO," with R. W. Parry, C. E. Mordman, and J. C. Carter, Adv. Chem. Series Jj2, 302-11 (196 V). "The Synthesis and Characterization of Cyclopentadienyl Derivatives, of Uranium and Thorium," with M. Dubeck, Inorg. Chem. 3, 16^8-50 (196^). "Lead and Lead-210 in Rai.m7a.ter," with R. B. Holtzman and H. F. Lucas, Jr* } Nature 21^, 353-55 (1967). "The Sources of Lead in Perennial Rye Grass and Radishes" with R. Dedolph, R. B. Holtzman and H. Lucas, Jr. Presented at the American Chemical Society Spring Meeting, Minneapolis, Minn. April 1969. "Air as a Source of Lead in Edible Crops" presented to the American Chemical Society in Spring!Meeting, Minneapolis, Minn. April 1969. EPI 000301 Ucyenber 1 , 3570 7 --i .by' . i'-.v. N11667.01