GEORGIA INSTITUTE OF TECHNOLOGY Engineering Experiment Station 0 PROJECT INITIATION

Date:

Project Title: The D and the L Isomers of 00 P"-DDT

Project No.: B - 378

Project Director: Dr. R. D. Kihrough, Jr.

Sponsor: Public Health Service . . 1970 . Effective PCC9439r, XID 1969, ...... Estimated to run until: . • . Novembey

Type Agreement: G111:1t. 1?0... 5P92:31-.03 ...... Amount;nt•. $ 11095*

Reports: Terminal Progress Report due within six months after completion - ten copies.

Contact Person: Dr. Otto A. Bessey, Chief Program, Dcv. and Eval. Branch Natl. Inst. of Env. 111th. Sci. National Institutes of Health U. S. Public Health Service Bethesda, Maryland 20014

*Excludes $129 to be transferred from unexpended balance of 13.61. Cost sharing requires additional *780; Cr.. panion Account =.:o. E-6o0-9144

Assigned to .... is41C1e4r .4-91.031 e4 PPierlce9 ..... Division

COPIES TO:

❑ Project Director ❑ Photographic Laboratory ❑ Director ❑ Research Security Officer ❑ Associate Director ❑ Accounting ❑ Assistant Directoqs) a Purchasing ❑ Division Chiefs ❑ Report Section ❑ Branch Head ff.Library 0 General Office Services ❑ Rich Electronic Computer Center Engineering Design Services

Itimp

GEORGIA INSTITUTE OF TECHNOLOGY Engineering Experiment Station

PROJECT TERMINATION

Date 9/9/71

PROJECT TITLE: The D and the L Isomers of 0,P'-DDT

PROJECT NO:

PROJECT DIRECTOR: Dr. R. D. Eimbroughs Jr.

SPONSOR: Public Health Service

TERMINATION EFFECTIVE: 5/31/71

CHARGES SHOULD CLEAR ACCOUNTING BY: 7/31/71

Oblications Bemaininal Final Report - due 11/3/71 Dr. KintbrouGh) Final Patent Report - /WU CE111.)

Nuclear & Diological Sciences Division

COPIES TO:

Project Director General Office Services Director Photographic Laboratory Associate Director Purchasing Assistant Directors Report Sectio Division Chief Library Branch Head Security Accounting Rich Electronic Computer Center Engineering Design Serviees 413:1- IC:UM tGl- X _it, INT C3 •X-• X WILT UV eCe 'X" M 43:E3E /41-1011 X. C)11S1--Se- EMINENT STATION 225 North Avenue. Northwest Atlanta, Georgia 3033.2

November 3, 1971

Division of Research Grants National Institutes of Health Bethesda, Maryland 20014

Gentlemen:

Enclosed is the final report for project ES 00281. If further information is needed, please let me know.

Sincerely,

Raymond D. Kimbrough, Jr. Senior Research Chemist

RDK:lsg Enclosure Final Report: The D and the L Isomers of o,p -DDT (Department of Health, Education, and Welfare, Public Health Service, Grant Number ES00281)

Raymond D. Kimbrough, Jr. Nuclear and Biological Sciences Division Engineering Experiment Station Georgia Institute of Technology Atlanta, Georgia 30332

I. Reaction Pathways for the Synthesis of the Title Compounds

1. The simplest method of resolving the D,L mixture of ap t -DDT into the pure optical isomers would be by chromatography on a column of an opti- cally active material. The interaction of the D isomer and of the L isomer in the D,L mixture of ap t -DDT with cellulose, which has a D configuration, would be different and in many cases such a procedure has been used to separate the components of a D,L mixture.

In the case of the D,L mixture of ap t -DDT, chromatography on cellulose, and on carboxymethylcellulose,did not produce any optical activity in the effluent. The reason for this is assumed to be that the very slight inter- action of the hydrophobic DDT and the hydrophillic cellulose was not adequate to produce any resolution.

2. The resolution of the D,L mixture of o,p'-DDT could be effected by synthesizing a molecule containing an amino group which could be used for. diastereomeric salt formation with an optically active acid such as d-10- camphorsulfonic acid. After the mixture is resolved, the amine could be diazotized and replaced by hydrogen or by a .

Synthesizing such an amine or acetamido compound using a synthetic se- quence similar to the . one employed in the usual synthesis of DDT (chloral and

1 chlorobenzene in sulfuric acid or an ce-trichloromethly-p-chloro-benzyl and chlorobenzene in the presence of sulfuric acid or other Lewis acid) did not give the desired product. A suitable diamino compound was obtainable 1 2 from the reduction of the dinitration product of o,p'-DDT. ' The diazoti- zation of this diamine proceeded with dehydrohalogenation, equation 1, which would destroy any optical activity that had been obtained in the compound before diazotization. This dehydrohalogenation occurred with diazotization under all conditions which produced diazotization.

diazotizatiodiazotization> CC1 3 -CHAr 2 -> CC1 2 = CAr 2 (1) ik asymmetric carbon

3. The resolution of the D,L mixture could be achieved by sulfonation of the aromatic rings, resolution of the diasteromeric sulfonate salt of an optically active base such as brucine, and subsequent removal of the sulfo- nate group. A disulfonic acid from both p,p'-DDT and o,p'-DDT was prepared,

I and II, by reaction with sulfuric acid containing 10% sulfur trioxide at 0 70 C. The analytical and spectral, IR and nmr, data on these compounds were consistent with their structures. The positions for substitution of the aro- 1 matic rings would be the same in sulfonation as in the nitration. SO H 3 Cl 1 CC1 -CH- 0 H

SO H 3 Cl

2 e The disulfonation of.I and II did not occur under the conditions usually used for this reaction, boiling 50% sulfuric acid. This reaction was attempted under a variety of conditions of higher temperature and both higher and lower concentration of acid. In every instance, dehydrohalogenation, equation 1, occurred before desulfonation. Hence, any optical activity would be lost on dXsulfonation.

The sulfonates are new compounds. Their chemical and biological proper- ties are discussed below.

4. pentachloride is known to replace a hydroxyl group with a chlorine and a with two chlorine. The possibility of con- verting o-chlorophenyl-p-chlorophenyl—acetic acid, o,p'-DDA, which can be resolved (see below) to o,p'-DDT with phosphorus pentachloride (replacement of the hydroxyl of the carboxyl with a chlorine and the carbonyl of the car- boxyl with two chlorine) was attempted_. This conversion did not take place, but a new reaction of phosphorus pentachloride was obtained. These results are described in "Phosphorus Pentachloride for the Replacement of Benzylic

Hydrogen with Chlorine."

3 [Reprinted from the Journal of Organic Chemistry, 34, 3655 (1969).] Copyright 1969 by the American Chemical Society and reprinted by permission of the copyright owner.

Phosphorus Pentachloride for the Replacement butylbenzophenone and phosphorus pentachloride to of Benzylic Hydrogen with Chlorine' 185 ° , giving

RAYMOND D. KIMBROUGH, JR. AND ROYCE N. BRAA1LETT 0 Nuclear and Biological Sciences Division, Engineering Experiment Station, Georgia Institute of Technology, Atlanta, Georgia 30332

Received April 10, 1969

Phosphorus pentachloride has been widely used to re- place the hydroxyl groups of and carboxylic acids to give alkyl and acyl chlorides, and to In the present work, it has been found that benzylic replace the carbonyl groups of aldehydes and ketones to hydrogen is replaced by chlorine when the material is give dichlorides. 2 heated with phosphorus pentachloride. The other prod- In specific cases, phosphorus pentachloride is known ucts of the reaction are presumably phosphorus tri- to replace carbon-bound hydrogen. Homophthalic and . acid, I, with phosphorus pentachloride gives a series of The replacement of benzylic hydrogen may proceed by way of a free radical reaction pathway. The first step might be dissociation of the phosphorus pentachlo- CH,COOH ride to and chlorine, a reaction known to occur at elevated temperature.' The chlorine COOH 0 would then react at the benzylic position in the familiar II manner.8 Cl, Toluene was converted to benzyl chloride by re- CCI,C0C1 fluxing with phosphorus pentachloride in almost quan- titative yield. With excess phosphorus pentachloride, Cl2 COC1 benzylidene chloride was obtained along with benzyl III IV chloride and unreacted phosphorus pentachloride. Thus the replacement of successive benzylic hydrogen products including II, III, and IV, where replacement seems to be increasingly difficult. . of carbon-bound hydrogen with chlorine has occurred. 3 When diphenylinethane was heated with 1 mol of Phenyl malonate, V, and phosphorus pentachloride phosphorus pentachloride, benzhydryl chloride was give a mixture of products including VI and VII where formed. Excess phosphorus pentachloride gave di- replacement of carbon-bound hydrogen by chlorine has phenylmethylene chloride. was occurred.' converted to triphenylmethyl chloride in 93% yield by phosphorus pentachloride at 140-150°.

0 0 0 0 0 .0 (1) This research was supported by Grant No. ES 00251 from the National II II II II II II 6II5 Institutes of Health, U. S. Public Health Service. C61150CCH•C0C CICCC1,CC1 C8H5OCCCI,COC 6116 V VI VII (2) R. B. Wagner and If. D. Zook, "Synthetic Organic Chemistry," John Wiley & Sons, Inc., New York, N. Y., 1953, pp 91, 105, and 546. (3) W. Davies and H. G. Poole, J. Chem. Soc., 1616 (1928). (9) V.I. Shevchenko and A. V. Kirsanov, Zh. Obshch. Khim., 35, 713 (1965). (5) B. Fell, Angetv. Chem., 77, 506 (1965). Chlorination of hydrocarbons, specifically the chlo- (6) W. Theilacker and F. Boelsing, ibid., 71, 672 (1959). rination of heptane to give heptyl chloride, with phos- (7) A. F. Holleman and E. Wiberg, "Anorganische Chemie," 32nd ed, phorus pentachloride and peroxides has been reported.' Walter de Gruyter & Co., Berlin, 1953, p 254. (8) D. J. Cram and G. S. Hammond, "Organic Chemistry," Second ed, An aromatic hydrogen is replaced by heating 2,2'-di-t- McGraw—Hill Book Co. Inc., New York, N. Y., 1961, p 523. 4

3656 NOTES - The Journal of Organic Chemistry

TABLE I REACTIONS OF PHOSPHORUS PENTACHLORIDE Mol•of PCla per mol of starting Conditions, Yield, Mp or Lit. mp Starting material material °C, (hr) Purification Product % bp, °C (mm) or bp, 'C Toluene 0.5 Reflux, Distillation Benzyl chloride' 97b 68-70 179' 110 (12) (40) Toluene 2 Sealed tube Distillation Benzylidene 38 105-107 214' 140-150 chloride' (12) (18) Diphenylmethane 1 140-150 Distillation Benzhydryl 78 160-162 161-162d (18) chloride. (12) Diphenylmethane 3 140-150 Distillation Diphenyhnethy- 90 170-173 172' (18) lene chloride' (12) Triphenylmethane 2 140-150 Recrystallization, Triphenylmethyl. 93 111-113 113-1141 (18) -hexane! chloride' Diphenylacetyl 2 140-150 Recrystallization, Diphenylchloro- 85 49-51 * 507. chloride" (18) pentane acetyl chloride' 1,1,1-Trichloro- 2 Sealed tube Recrystallization, 1,1,1,2-Tetra- 88 90-92 90-91° 2,2-bis(p-chloro- 180-200 ethanol chloro-2,2-bis- phenyl)ethane (18) (p-chloropheny1)- (p,p'-DDT) ethaneg The infrared spectrum was identical with that of the authentic material. 6 This yield was based on unrecovered toluene. Based on phosphorus pentachloride, it was 42%. N. H. Lange, "Handbook of Chemistry," 7th ed, Handbook Publishers, Inc., Sandusky, Ohio, 1949. a H. Gilman and J. E. Kirby, J. Amu. Chem_ Soc., 48, 1735 (1926). e L. Gattermann and H. Schulze, Ber., 29, 2944 (1896). L. Gattermann and H. Wieland, "Die Praxis des organischen Chemikers," 39th ed, Walter de Gruyther & Co., Berlin, 1959, p 298.

g The of this material was not depressed by mixture with the authentic material. " H. Staudinger, Ber., 44, 1619 (1911). The identity of this material was established by conversion into ethyl diphenylchloroacetate, mp 42-44° (lit.' mp 43-44°), by solu- tion in ethanol, and by conversion into mandelic acid, mp and mmp 117-118°, by reflux in potassium carbonate solution. 7 H. Bic- kel, Ber., 22, 1539 (1889).

Diphenylacetyl chloride, IX, heated with excess phos- to an ethylenic double bond is known." The chlorina- phorus pentachloride gave diphenylchloroacetyl chlo- tion of the other compounds in Table I cannot involve ride, X, in 85% yield. such an olefinic intermediate. The several compounds mentioned above in which benzylic hydrogen is replaced by chlorine on heating (C8115)2LOC1 CC13C(C6114CO2 with phosphorus pentachloride suggest that this is a IX, X = H XI, X = H general reaction. X, X = Cl XII, X = Cl When 1,1,1-trichloro:2,2-bis(p-chlorophenyl)ethane, Experimental Section the insecticide p,p'-DDT, XI, was heated with phos- Replacement of Benzylic Hydrogen with Chlorine USing Phos- phorus pentachloride at 180-200° for 18 hr, an 88% phorus Pentachloride.—A reaction typical of those outlined in yield of 1,1,1,2-tetrachloro-2,2-bis(p-chloropheny1)- Table I is described for the preparation of XII. A mixture of ethane, XII, was obtained. The preparation of XII is 5.0 g (0.013 mol) of XI and 7.2 g (0.026 mol) of phosphorus usually from XI in two steps, the dehydrochlorination pentachloride was sealed in a,40-m1 glass tube and heated at 180-200° for 18 hr. The cooled reaction mixture was added to of XI with base to give 1,1-dichloro-2,2-bis(p-chloro- ice water and stirred until the excess phosphorus pentachloride phenyllethylene, XIII, followed by the addition of and the phosphorus trichloride were decomposed. The in- chlorine to the ethylenic double bond of XIII to give soluble material was taken up in ether and the ether layer was XII.' dried with magnesium sulfate and- evaporated. The residue was recrystallized from ethanol: yield 4.5 g (88%); mp 90-92° C12C=C(C6II.,C1)2 mp 91-92°). XIII Registry No.—Phosphorus pentachloride, 10026- To test the possibility that XIII might be an inter- 13-8; toluene, 108-88-3; diphenylmethane, 101-81-5; mediate in our conversion of XI to XII with phos- triphenylmethane, 519-73-3; IX, 1871-76-7; XI, phorus pentachloride, XIII was heated with phosphorus 50-29-3. • pentachloride. An almost quantitative yield of XII was obtained. Hence all that can be said is that XIII (9) 0. Crumrnitt, A. Buck, and A. Jenkins, J. Amer. Chem. Soc., 67, may be an intermediate in this reaction. The use of 155 (19-15). phosphorus pentachloride for the addition of chlorine (10) L. Spiegler and J. M. Tinker, ibid., 61, 990 (1939).

5

II. o-Chlorophenyl-p-chlorophenylacetic Acid (o,p'-DDA) 3 The D,L mixture of o,p'-DDA can be resolved as the brucine salt by

chromatography on silica gel with 80% aqueous ethanol. The resolved o,p'-DDA

can be recovered by evaporating the effluent, dissolving the residue in water, and acidifying with 5% hydrochloric acid. The first material which

came off the column had a less negative rotation than the original salt while the last fraction had a more negative rotation. The specific rotation

0 of the acid recovered from the first and last fractions was + 1.1 and - 1.4 .

III. Properties of the Disulfonic Acid Derived from DDT

1. The disulfonic acid derived from DDT, I, is very water soluble, is

less soluble in most organic solvents, and is insoluble in nonoxygenated or-

ganic solvents. The salts of this disulfonic acid are very water soluble

and insoluble in all organic solvents. This is the first water-soluble, fat-

insoluble derivative of DDT to have been made.

The results of acute toxicity studies in both insects (mosquito larvae)

and mammals (rats) and the results of long-term feeding studies in mammals

(rats) indicate that the material is nontoxic, that is about 1% as toxic as

DDT. Preliminary metabolism studies indicate that the material is excreted

rapidly and essentially unchanged. (The possibility that the disulfonic

acid may be partially or completely dehydrohalogenated to III has not been

eliminated.)

Cl

III

6 2. The sodium salt of the disulfonic acid was dissolved in water and

at a concentration of 100 times the LD for mosquito larvae, no kills were 50 observed.

The sodium salt of the disulfonic acid in water solution was given

orally to rats. At 50 times the LD for DDT two out of two rats survived. 50 At lop times the LD50 for DDT one of two rats survived. (The LD 50 for DDT

is about 250 mg/kg, so that 50 times this is 12.5 g/kg and 100 times is 25

g/kg.)

Rats were fed diets of up to 27 the sodium salt of the disulfonic acid

for 3 months with no apparent effects. This is about 2 g per kg rat weight per day, approximately 100 times the 90 day LD 50 for rats. ate weight gft4-101 eiThe rats fed the salt gained weight at the same rate as the controls and

there were no abnormal findings on autopsy.

3. The complete lack of toxicity of the disulfonic acid derived from

DDT is explained by its . DDT is water insoluble and fat soluble.

It exerts its toxic action in the neural tissue which are lipids. The di-

sulfonic acid salt is totally insoluble in fats and hence does not enter the

lipid tissue. Because of its solubility characteristics, DDT is stored in

the lipid tissue and accumulates in the animal. The disulfonic acid salt is excreted rapidly and is not stored, presumably because of its solubility.

IV. A Sulfonate Ester Related to DDT

1. As mentioned above, sulfonic acid salts are water-soluble and fat-

insoluble. Hence these substances would not accumulate in the lipid tissue of plants and animals and would not tend to be concentrated in the food chain

as are substances like DDT which are fat-soluble and water-insoluble. The

studies discussed above indicate that sulfonic acid salts do not have the

7 toxicity associated with the structurally related pesticide, presumably be- cause of the solubility differences in the two substances.

The alkyl esters of sulfonic acids are fat-soluble and water-insoluble so that alkyl sulfonate esters of structure related to chlorinated hydrocarbon pesticides like DDT will have the same solubility properties as the pesticide and hence may have similar pesticidal properties. Under conditions in the field, the fat-soluble, water-insoluble alkyl sulfonate esters are slowly hydrolyzed to water-soluble, fat-insoluble sulfonic acid salts, so that these substances would not be persistent and would not accumulate in the food chain.

Ideally, these sulfonate esters related to chlorinated hydrocarbon pes- ticides like DDT, would have the desirable qualities of the chlorinated hydrocarbon pesticides, that is, high toxicity to insects and low toxicity to higher animals, but, unlike the chlorinated hydrocarbons, would not be per- sistent and would not accumulate in the food chain.

2. A sulfonate ester related to DDT, IV, was synthesized by the reaction sequence given in equation 2. The final product IV had the analytical and spectral properties expected for a material of its structure. IV was soluble in organic solvents and insoluble in water.

H SO H SO •S0 2 4 2 4 3, CC1 -CHO + C H C1 > CC1 -CH-C H 3 6 5 3 6 70 °C C H 6 5 PC1 CHOH 5 3 SO 3H > CC1 -CH SO C1 2

SO H SO Cl 3 2

(2)

SO CH 3 3 IV 8 3 . Preliminary studies of the toxicity of IV indicated a slight tox- icity to mosquito larvae, far below the toxicity of DDT; however, this work is unfinished.

References

1. J. Forrest, 0. Stephenson, and W. A. Waters, J. Chem. Soc., 1946, 338.

H. L. Haller, et al., J. Am. Chem. Soc.„67, 1591 (1945).

2. 0. G. Backeber and J. L. C. Marais, J. Chem. Soc., 1945, 803.

3. Org. Syn., col. vol. 3, 270.

A manuscript for publication on the disulfonic acids related to DDT the synthesis and toxic properties, is presently in preparation.

9