Document KdeZn9Kp0j6kKxxmMaOab0z0

Castleman File: American Petroleum Institute w/c = with cover letter or memo If DATE = 0, undated CD-ROM Document #:API -7 i DATE __ published article from trade journal __ published advertisements __ newspaper article __ published government report __ government inspection results unpublished or internal report unpublished presentation from conference letter memorandum industry warning labels industry sales literature Industry recommended practices meeting minutes (with attachments) membership list BC notes eetterino LAiORAToRT lSE OF MEDIC,NE-IDER ((INNATI 0H|Q UNIVERSITY OF CINCINNATI DEPARTMENT Of PREVENTIVE MEDICINE AND INDUSTRIAL Health June 26, 1953 Mr. d, V. Stroop, Director Department of Technical Services American Petroleum Institute 50 West 50th Street New York City 20 Dear Mr. Stroop: I herewith acknowledge with thanks receipt of the following checks, covering expenditures for the year, beginning June 30, 1953: Investigation of skin physiology Investigation of fluorine compounds $ 5,000.00 2,500.00 ef API 05619 i. American Petroleum Institute 50 WEST 50th STREET NEW YORK SO, N. Y. Mas 30, 1953 Board of Dlraetora UbiYarsityof Cincinnati Cincinnati 19, Ohio ~ Oantla--m Ms --silsartLrja la authority to centlaws sort on lanrastijntiaa of tte tanluity sad nods of notion of ocrtala patrol** jrodust* for tte jmae msIIhe fans 30, 195V, according to tho tog-- of tho agraanaut datad January 8, 19*6, sad on tte basis of a awl-- tedgst for rparstlng aapanaas of #78,283. typa racalpt of four approval of tha continuation of tte in>>t with this Mattel todgat -- teail pay yea tte a-- of **>,000. tts telsurra of 136^ tea tte 11--ipateal teLuwa in tte fad an Jten 38, >953* tett te fate <a 1--1117% 395V. It la --laratoait that mj telawa in tte fund on June 30, 195V *111 te ratvraad to tte h--rlaan Patrols-- Institute, and that no anasa orar tte e--t of tte tedgst is to te efeligatad or spool without written --thcriaation. Awmti fsxaouui narrrun Acoaptad ate Approwtet lanviRsrn 0w ctbcubati Rnrougb its Board of Uraators UNIVERSITY OF CINCINNATI DEPARTMENT OP PREVENTIVE MEDICINE AND INDUSTRIAL HEALTH CASL! AODRESE KETLAI, CINCINNATI TELEPHONE. CAhtoc UK Mr. D. V. Stroop, Director Department of Technical Services American Petroleum Institute 0 West 50th Street New York City 20 Dear Mr. Strobpj I herewith acknowledge with thanks receipt of American Petroleum Institute's check in the amount of $i|0,000.00, covering expenditures to be made on their behalf. Very truly yours. ef API 05621 UNIVERSITY OF CINCINNATI DEPARTMENT OF PREVENTIVE MEDICINE AND INDUSTRIAL HEALTH August 26, 1953 CARLE ADDRESS: KETLAB, CINCINNATI TELEPHONE: CAPITOL 1414 Mr. D. V. Stroop, Director Department of Technical Services American Petroleum Institute 50 West 50th Street New York City 20 Dear Mr. Stroop: . I hasten to correct an error made in my letter of August 25. The amount of American Petroleum Institute's check of July, 1953 should have read $40, 000. 00. Very truly yours, Dr. Kehoe ef API 05622 API 05623 J -(RIND LASORATORY * 0, medicine-*^ avemue <a,i..oh.o UNIVERSITY OF CINCINNATI DEPARTMENT Or PREVENTIVE MEDICINE AND INDUSTRIAL HEALTH August 25, 195 3 CAS LI ADDRESS: RET LAB. CINCINNATI TELEPHONE: CAPITOL MI4 Mr. D. V. Stroop, Director Department of Technical Services American Petroleum Institute 50 West 50th Street New Ydrk City 20 Dear Mr- Stroop: 4-r I am sending you herewith, for your information, a statement of expenditures made'on behalf of the American Petroleum Institute for the second quarter of 1953. (This statement does not include American Petroleum Institute's check in the amount of $LUi UUU. UD~ received on July 9, 1953.) I believe that this statement will be self-explanatory, but if you have any questions or comments, Doctor Kehoe would appreciate your bringing them to his attention. Very truly yours, ef Enc. -- .r -....... E. R. Fortlage/Secretary to Dr. Kehoe API 05624 UNIVERSITY OF CINCINNATI KETTERING LABORATORY I ACCOUNT OF__ THE AMERICAN PETROLEUM TWSTTPTTnc I FOR 2nd Quarter 1953 SALARIES (Based on Proportion of Time Actually Spent on Project) Direct Salaries .1U6.45*.4&. Indirect Salaries Histopatholofical Preparation------------------------ __ .25&X.Q.Q- Other Services....................................................... ...........21 .954.75 miscellaneous expense Purchase of Animals--------------------------------------------------------------------- 2j8.k0 Special Laboratory Supplies....................... .........................................3.4.6.20. Trml.7.12 jjjL Overhead (Proportion of Heat, Gas, Electricity, Steam, Telephone, Ceneral Laboratory Supplies, Postage, Annuities, Pensions, Maintenance, etc------ 7jg0P.BA flr55Q.QQ Balance Avaiabie for Further Work TOTAL ................. st End of27,OH.10 Balance Due Kettering Laboratory at End of. Receipts Expenditures 2nd Quarter 1953 Balance Available for Further Work at End of._______________________________ Balance Due Kettering Laboratory at End of. 2nd .-Q.uar.tflr. 301494172 3,450.62 (|* IVT LAI. SN 112 30,494.72 API 05625 EXPLANATORY NOTES CN TABULAR SUMMARY OP BIOLOGICAL EXPERIMENTS September 1, 1Q53 degree of confidence put by the investigators in the apparent potency of the material as indicated by the rate of induction of tumors. In the oases in which it was felt that the state of health 0f the animals, as measured by growth curves and survival rates, was not sufficiently good, the tabulated potency, PMC, has been set down in parentheses. Normally, any given sample of oil was applied to two or more groups of mice, each group containing 6 to 10 animals. If there were significant differences in the apparent state of health of the different groups, the "Average Latent Period for Tumor In duction" was calculated using the data on times of appearance of tumors from those groups in which the growth and rate of survival approached that of control animals. The figures associated with this procedure have been indicated in the columns on "Original Number of Mice" and "Pinal Effective Number of Mice" In the Tables, in that the total numbers of animals used initially and the corres ponding effective numbers are indicated in parentheses, while the numbers used in the calculations are shown directly above. The "Average Latent Periods" have been recalculated by a slightly modified method and are shown in the Table with the 5 percent fiducial limits, which signify that the statistical probability is only 1 in 20 that the true mean time of tumor induction lies beyond these limits. When there were indications. API 05626 -2- from the growth or mortality data, that the experimental procedure was affecting the health of the animals to some extent but not so severely as to render the results ambiguous, the Average Latent Period was put in parentheses, but the value of the potency, PMC, was not so qualified. Summarizing, the experiments have been classified into three grades indicated in the Tables as follows: GRADE , _________ AVERAGE LATENT RELATIVE PERIOD FOR CARCINOGENIC TUMOR INDUCTION POTENCY.?Mf! A (normal growth and survival) B (some symptoms of abnormal conditions, but reliable estimate of potency possible) X (reliability of results uncertain) (____ ) (_____ ) ____ (_____ ) API 05627 For Information Only - Not for Publication API Researon frojecc MC-x Atf*rr \ TABULAR SUMMARY OF CURRENT AND COMPLETED BIOLOGICAL EXPERIMENTS September 1, 1953 The Kettering Laboratory in the Department of Preventive Medicine and Industrial Health College of Medicine University of Cincinnati Cincinnati, Ohio API 05628 INDEX Index to Processes index to Table I Table I - Experiments on Refinery Streams and Fractions Thereof Table II - Experiments on Synthetic Carcinogens and Non-accelerating Solvents Concentration Standards with Methylcholanthrene in Benzene Experiments Involving the Application of Solutions of Methylcholanthrene in Sec,,-amylbenzene to the Skin of Mice Experiments Involving the Application of Solutions of Methylcholanthrene in Various Solvents te the Skin of Mice Control Experiments with Various Solvents Table III - Experiments on Acceleration of Carcino genesis by Specific Solvents Experiments Involving the Application of Solutions of Synthetic Carcinogens in Dodecylbenzene to the Skin of Mice Experiments Involving the Application of the Accelerating Solvent Prior to or After the Application of the Carcinogen Experiments Involving the Application of Solutions of Benzpyrene in Various Solvents to the Skin of C3H Mice Experiments to Determine Whether Certain Materials Have Irritational Properties of the Type Evidenced by Dodecylbenzene Table IV - Experiments Involving a Limited Number of Applications of Carcinogenic Materials Table V - Experiments on the Retardation of Tumor Formation by Washing Page Humber i 2 6 26 30 31 32 33 35 37 43 46 47 API 05629 INDEX TO PROCESSES Process jj^n-catslytic cracking of virgin gas oil . . Non-catalytic cracking of catalytic gas oil Steam cracking Viscosity breaking pubbs coking 3atch coking Delayed .`.oking Naphtha . ^forming Polyforming yiydroforuiing Thermofor catalytic cracking (T.C.C.) Fluid catalytic cracking (F.CiC.) Houdry Cycloversion ' Solvent extraction . Fractional distillation Straight run distillation Lubricating oil VJax pressing MEK - Benzol solvent dewaxing Filtration dewaxing Desulfurization by catalytic hydrogenation Acid treating Viscosity effects Retorting of oil shale Puel oil blending Diels-Alder reaction Solvent extraction with concentrated HaS04 Air oxidation Chromat ography Appendix Page Number 6 6 9 9 9 10 10 11 11 11 11 . 13 16 ' 16 ' 17 17,19,22,23,25 18 19 20 20 20 20 20 20 20 21 22,23 23 21). 24 API 05630 cxioil>cxj0i i 3 0 ooocx)Oocx>cocx>cjquj jj--j a vn.-fr v_> tv, v-*i i "i i t i t i i >x API saopi number* -2INDEX TO TABLE I Process Straight run distillation Non-catalytic cracking of virgin gas oil T. C. C. P. C. C. Straight run distillation Non-catalytic cracking of virgin gas oil T. G. C. F. C. C. Filtration dewaxing Dilution of No. 8 Chromatography . Fractional Of No. 8 of No. 8 of No. 8 of No. 8 of No. 8 of No. 8 Diels-Alder reaction of No. 8 of No. 8 Chromatography of No. 8 Dilution of No, 8 of No. 8 Chromatography of No. 8 Solvent extraction with concentrated HaS04 Solvent extraction with concentrated HaSOa Dilution of No, 8 Diels-Alder reaction Chr omat ography T. C. C. T. C. C. Houdry F. C. C. Chromatography Distillation (1 plate vacuum) Distillation (1 plate vacuum) Distillation (1 plate vacuum) Fractionation Polyforming solvent extraction Solvent extraction Viscosity breaking Naphtha reforming Non-catalytic cracking of catalytic gas oil Dilution of No. 20 Desulfurization by catalytic hydrogenation Cycloversion Appendix Page Number Id 6 11 13 18 b 12 111 20 14 22 22 22 22 22 22 22 22 22 24 22 2? 14 24 22 23 23 24 12 12 16 15 24 25 25 25 23 11 17 17 9 11 b b 20 lb x .cc i Ci Bi Ci > Li C iu o io o > ixi u io cu o a *-- --i--o * j -* *O 'D---tg >----> >r--v;-- i--iv j r--ivj--* --iru r \ju o u \ rv*o - r\o v> fvjh rv. * Experiments which are complete, including micropathology, are indicated by an asterisk in the body of the tables. API 0563! API Sample wmnber -3INDEX TO TABLE I Process C C Solvent extraction tilth concentrated H,SO* Solvent extraction with concentrated He304 Non-catalytic cracking of catalytic gas oil Distillation . Dilution of No. 25 P. C. C. Solvent extraction Delayed coking Catalytic feed to non-catalytic cracking Non-catalytic cracking of catalytic gas oil T. C. C. Wax pressing Delayed coking Houdry Non-catalytic cracking of catalytic gas oil T. C. C. . _ Polyforming Catalytic feed to non-catalytic cracking Non-catalytic cracking of catalytic gas oil T. C. C. . Delayed coking F C C. F. C. C. MEK - Benzol solvent dewaxing F. C. C. Non-catalytic cracking of catalytic gas oil F. C C Non-catalytic cracking of catalytic gas oil F. C. C. Hydroforming Houdry Houdry Dubbs coking Dubbs coking Catalytic feed to non-catalytic cracking Non-catalytic cracking of catalytic gas oil Straight run distillation Wax pressing Naphtha reforming Non-catalytic cracking of virgin gas oil T. C. C. Non-catalytic cracking of catalytic gas oil Acia treating Houdry Non-catalytic cracking of catalytic gas oil Non-catalytic cracking of virgin gas oil Viscosity breaking Viscosity breaking F C. C. Polyformlng Solvent extraction Appendix Page Number 7.13 23 23 7 17 17 15 17 10 7 7 12 20 10 7,16 7 12 11 7 7 12 11 15 15 20 7.15 7 8.13 8 15 11 16 16 9 9 Q 8 18 20 11 6 11 8 20 8.16 8 6 9 9 1? 11 17 API 05632 i 4 Y J API I sample I number 71 , 77H1-2 72 73 1L -4- INDEX TO TABLE I Process T. C. C. Chromatography Unromatography won-catalytic cracking of virgin gas oil Ft C. C. jeiayed coking solvent extraction .,apntha reforming hydroforming i C C Straight run distillation Houory mx (w e U* Air Oxidation air oxidation Delayed Coking i r n steam cracking natch coking F. C. C. Ft C. Delayed coking Catalytic feed to non-catalytic cracking Non-catalytic cracking of catalytic gas oil Non-catalytic cracking of catalytic gas oil Non-catalytic cracking of virgin gas oil Delayed coking Delayed coking Delayed coking Lubricating oil Lubricating oil Lubricating oil rubricating oil x a V- e X. C. C. ' Delayed coking Ft C. C . Straight run distillation Non-catalytic cracking of virgin gas oil T. C. C. fractional distillation Fractions? Distillation fractional distillation Fractional distillation Dilution of No. 111-4 Dilution of 'J'j. 111-4 Fractional distillation Fractional distillation Fuel oil olending Appendix Page Number 12 2k 24 b 15 10 17 11 11 12 lb lb 12 2k 24 10 15 Q 10 13 13 10 9,1o3 9 6 10 10 10 19 19 19 19 13 13 10 12 19 19 19 19 19 19 19 19 21 API 05633 1. API Sample wumber llii llll-I 11$ 116 118 119 -5- INDEX TO TABLE I Process Straight run distillation Viscosity effects Straight run distillation Retorting of oil shale Fuel oil Blending Fuel oil Blending Appendix Page Number 18,20 20 18 20 21 21 - 6- T TABLE I SKIN PAINTING EXPERIMENTS ON HIGH BOILING SAMPLES A IALITI CAL DA T A DISTILLATION . ,2mm. corr, to 76Qna, PROCESS API SAMPLE NUMBER PRODUCT GRAV ITY VISCOSITY 750 <75cF 925 >925 E.P, ($) (*) ($) (P.) Non-catalytic cracking of virgin gas oil n 2 6 n 59 65 n 72 it 9k !t 108 Cracked sidestream Cracked residuum Cracked residuum Cracked residuum Cracked residuum Cracked residuum Cracked residuum 21.2 33/100 ssu < 5.2 33 26 1+1 - 8.5 33/100 ssu 32 6.6 163.5/100 1+3.8 29.9 26.3 ssu 9.6 37.1/122 SSF 35 37.5 27.5 10.1). 31.7/122 35 1+8 17 SSF 2.3 1530/130 ssu 32 25.5 1+1.9 - <<41 1 % Non-catalytic 20 Cracked cracking of residuum catalytic gas oil if.7 132/122 5l 33 16 SSF Dilution of No. 20 20-1 $0% oil No.20, 50$ dodecylbenzene 1 i l! J API 05635 -6- TABLE I SKIN PAINTING EXPERIMENTS ON HIGH BOILING SAMPLES STRAIN OF Nil CE AND NUMBER OF A?I 3AJ1PL3 APPLICA TIONS jxjmblR, PER WEEK H DOSAGE PER . APPLICATION BIO LOG] C A L DA T A FINAL EFFEC ORIGINAL TIVE NUMBER NUM3ER OF OF MICE MICE AVERAGE MAXIMUM LATENT INCIDENCE PERIOD RELATIVE OF FOR TUMOR CARCINOGENIC TUMORS INDUCTION POTENCY, (T.I./wks.) (5$EL. in wks.) pMC ! 2 C3H 1 100 20 7l 0/75 - - 6 * CFW 3 100 59 C3H 3 100 65 C3H 3 100 72* C3H 3 100 9^ C3H 3 100 108 C3H 3 108 C3H 3 20 * CFW 3 20 C3H 3 100 5 100 100 20 (30) 20 18 (21) 11 20 9 20 15 6 (20) 5 (9) 20 7 (19) 20 20 7 6 (13) 15 ' 10 72/30 (76/30) 82/69 89/29 73/16 80/39 (67/39) 71/28 100/52 (92/52) 100/22 90/21 (25.2-3.5) (53.2-8.7) o.o8:-j wC (23.6-3.3) (o.i2:;W) (15.1-1.8) (0.P2+;|) (31.0-13.6) (O.O9:;08) (2^.015.6) - OO 0 +0r -1 0 (36.1*12.5) (15.oil.8) (0.15:;^) (17.4-2.1) - 20-1# CFW 3 100 20 17 100/25 (I6.li2.7) 00,15-.0^57 i Number alive after 63 weeks API 05636 "7" T ui ve TABLE I SKIN PAINTING EXPERIMENTS ON HIGH BOILING SAMPLES A N A L Y T I -C A L DATA DISTILLATI01 (2mm. corr to 76011c, PROCESS API SAMPLE NUMBER PRODUCT GRAV ITY <75cf VISCOSITY (*) 750 925 >925' (*) (F.l Non-catalytic cracking of catalytic gas oil (1 23 24 n 29 ft 30 ft 34 ti 35 tf 38 N 39 tt 44 If 45 --- -- " -- |1 PCC gas oil feed 26.7 Cracked residuum from No. 23 5-9 TCC gas oil feed 15.7 Cracked residuum from Nc. 29 8.5 Houdry gas oil feed 28.0 Cracked re s i duum from No. 34 2.7 Total cracking 16.1 charge; com bination unit Cracked residuum from No. 38 7.8 FCC decanted cil feed 15.2 Cracked residuum from Nc. 44 -1.7 55.5/100 73.5 20.6 ssu .9 636 51.9/210 32.3 34. b 33.1 ssu 70/100 73 27 ssu - 860 10/210 3h 38 26 SSP - 47.5/100 9 615 ssu 2320/100 *3 28 r * ssu 2333/100 32.2' 23.3 43.1 * LSU 177/122 31.0 19.8 48.4 SSF 241/100 31.1 54.2 3 3.6 0 J LSU 179/122 2o.i| 3 3 = a 39 SSF i API 05637 TABLE I SKIN PAINTING EXPERIMENTS ON HIGH BOILING SAMPLES r-- API r sample WuMBEF STRAIN OF MICE AND NUMBER OF APPLICA TIONS PER WEEK tOac MO rS0<Hi 0>H C00 Q0 J<ianti (mg J B I C L 0 0 I C A L DA T A FINAL AVERAGE EFFEC MAXIMUM LATENT ORIGINAI TIVE INCIDENCE PERIOD RELATIVE NUMBER NUMBER OF FOR TUMOR CARCINOGENIC OF OF TUMORS INDUCTION PCTENCY.- MICE MICE (To 10/weeks) (EL. in wksO PMC 23* CFW 3 100 20 20 23* C3H 2 100 20 13 100/19 91+ /2 8 12.51.3 (18.512.9) -25-:oi 2k* CFV7 3 100 19 . 18 100/15 (9.515) > 0.5 360 29* C3H 1 100 20 19 30* C3H 1 100 10 (20) 8 (13) 0 15 Ik* CFW 3 100 20 19 35 CFW 3 100 20 13 Q1+/30 100/33 (92/33) (23.3+3.?) (24.51+. 8) o.4o+# Jj1 "l4 0,5Ci''t'0?5 ' ' -. 81+/32 100/20 (23.2+3.3) (16,7+1.5) 0,nq+,"i -,02 Q.15+,02 38 CFW 3 100 19 13 62/58 (45.1112.6) (.o5:;g|) 39 CFW 3 100 29 18 56/21+ (21.9+2.7) = 1*4 C3H 1 100 45 C3H 1 100 20 10 (20) 16 8 (9) 91+/1+0 (31.1*3.8) (0.25l;gS> 88/41 (89/l+D (34.2+5.4) (o.ax*:) API 05638 TABLE I SKIN PAINTING EXPERIMENTS 017 HIGH BOILING SAMPLES A 17 A L Y T I C A I ITTtt DISTILLATION 2mm. corr. to 76 PROCESS API SAMPLE NUMBER PRODUCT GRAV 75o5 <750 925 >925( ITY VISCOSITY {%) (*) (*) Non-catalytic cracking of catalytic gas oil ft It ft 46 FCC gas oil feed 47 Cracked residuum from No. 46 54 Total crack ing charge 55 Cracked residuum from No. 54 61 Cracked residuum 63 Houdry gas oil feed 23.6 56.7/100 87.9 12.1 ssu 0 9.6 140/100 76 15.5 8.3 ssu 33-0 4-2 7.8/122 93.9 4.1 SSF 2.C 18.8/122 55.9 26.1 18 SSF 6.3 25.6 73.6/100 66.8 18.2 SSU 25.7/100 46.5 37.6 ssu 15 13.9 If 64 Cracked residuum 19.9 31.8/100 52.5 31.9 15.7 ssu from No. 63 API 05639 rd TABLE I SKIN PAINTIN'! EXPERIMENTS ON HIGH 30ILING SAMPLES - BIO L 0 G I CAL DA T A ....... - STRAIN % $ OF MICE i EH '.) API SAMPLE tfOMBEP AND number OF APPLICA TIONS 0Cj3 MaJ aO) C2hQ< PER WEEK (mg,) FINAL AVERAGE EFFEC MAXIMUM LATENT ORIGINAL TIVE INCIDENCE NUMBER NUMBER OF OF OF t TUMORS PERIOD RELATIVE FOR TUMOR CARCINOGENIC INDUCTION POTENCY, MICE MICE |T. I,/weeks) (50.L. in iik: > PMC iO ij.6* CF'.V 3 100 20 15 100/21*. (17.9*1=5) 0 l-! + o02 U^-.01 > 1*7 CFW 3 100 20 14 93/17 (13.Oil.6) 0"^l2? -+.=0160 22 54 CFW 3 100 20 16 IOO/36 (25.02.9) 0.07*.01 - 55* CFW 3 100 9 8 89/19 (13.3*2.2) (19) (15) (87/21) - 61 CFW 3 100 20 17 88/26 (15.2+3=8) - 63* C3H 3 100 20 18 89/26 (20.311.9) (0.14*.02) 63 C3H 3 100 63 C3H 2 100 12 U9) 20 12 (15) 18 100/19 (100/19) IOO/33 (15.7*2.0) (23.4*2.6) 0.19+.04 "^ 5 -i7-:o35 - 61** C3H 3 100 20 13 IOO/23 (16.8+1.9) (0-17Do2> API 05640 1 i * I 1I 4 iI -9- TABLE I SKIN PAINTING EXPERIMENTS ON HIGH 30ILING SAMPLES > A 1i A L Y T I mm"FT" DISTILLATION mm. corr. to 76c PROCESS API SAMPLE NUMBER PRODUCT GRAV 750 <75^ 925 >925c ITY VISCOSITY (*) (*) (%) Non-catalytic 91 FCC gas oil cracking of feed catalytic gas oil ft 92 Cracked residuum from No. 91 (f 93 Cracked residuum 20.1; 57.8/100 80,1 17.9 ssu 2.0 6.1 102.5/100 70.5 27 ssu 2.5 7.9 49.4/100 87 4.5 8.5 ssu Steam cracking 86 Cracked residuum 3.5 5000/1000 51.9 28.4 181 ssu Viscosity breaking ff ! Dubbs coking 18 Cracked residuum from No. 17 s 66 Cracked residuum 3.1 1123/210 23.7 24 ssu 52.3 6.5 18/210 22.1 29 48.9 SSF 67B Cracked side 15.6 stream from fractionator producing No. 66 81/100 84.6 10.7 4.7 SSU 528 Cracked sidestream 2 53 Blowdown oil 21.0 43.5/100 95.2 2.5 2.3 ssu 10.5 203/100 61.3 23.3 15.4 ssu 1 Distillation stopped when cracking occurred. a Peed to unit included catalytically cracked components. API 05641 ON 760m"u. \ if r- ,, i 0 830 9- I SKIN PAINTIN'j LX.i:_,ftIi.EN?fc> ON RICH 3CILI1.0 Sn.ii.PLLS "sTr.nIX OF MICE AND NUM3ER OF 4PI applicasAWPLE TIONS NUMBER PER V.EEK D 1 0 ij 0 a x -0 C-, m-j *w'l < rM FINAL EFFEC- CRIMINAL TIVE O CL, Q< NUMBER OF NUM3SR OF (mg.! MICE MICE CAL DA I A --1---------------------- AVERAGE MAXIMUM LATENT INCIDENCE PERIOD RELATIVE OF FOR TUMOR CARCiNCOENIC TUMORS INDUCTION POTENCY, (T. I, /weeks] ]5$EL. in wksl PMC 91* C3H 2 ICO 20 19 95/23 (16.4+2.1) (0,28"<'/) 5 5i 8801 9- n '92 C3H 2 100 93 C3K 3 100 T *3 (20) 20 11 (14) 18 91/23 (79/23) 94/40 (liJ.414.1) iC,33+,Hh ( 29 0 0 12 e 8 ) (o,o9!;5;) 36 C3H 1 100 18# CFW 3 100 10 (20) 10 8 (17) 100/36 (100/49) 9 56/30 (35.011,2) o,:^t3oi (30.1111.3) iC.0?,C3) 66 C3H 2 100 67 C3H 2 100 20 10 (20) 19 9 (13) 100/37 89/17 (77/17) 29,312,0 11,313.2 wr\ J 1.* -*aa 9 nw J'. - 1 ; , 26 ' " . -i 3 -l - 52 CFV.' 3 100 53 CFV.r 3 100 10 (20) 20 8 (15) 20 IOO/36 (87/36) 100/15 (18,92:7.9) 0 ti a. *rP.''7 1 10.211.2 0.531 ' API 05642 ui - 10 - TABLE I SKIN PAINTING EXPERIMENTS ON HIGH BOILING SAMPLES A N ALYT I C A L DATA DISTILLATIO] (2mm. corr. to PROCESS API SAMPLE NUMEER PRODUCT Batch coking 87 Cracked sidestream GRAV ITY 750 750 925 >925' VISCOSITY <*> ($) it) 14.9 500/1000 ssu Delayed coking 28 Cracked sidestream 15.3 361/100 ssu 42.4 ft 74 Total furnace 12.5 198/122 22.3 32.5 45.2 charge(includ SSP ing recycle) m 33 Cracked 16.8 131.4/100 60 38.1 1.9 sidestream ssu from No. 74- n 81+ Cracked 30.2 55/100 79 19.9 1.1 sidestream SSU n 90 Cracked 30.5 40.6/100 90.2 9.2 .6 sidestream SSU n 96 Cracked 33.2 43.7/100 77.7 22.2 .1 sidestream SSU ft 97 Total furnace 16.6 386/100 57.8 32.2 10 charge(includ ssu ing recycle) ft 98 Cracked sidestream 31.0 38.1/100 100 ssu from No. 97 #t 103 Cracked sidestream 25.2 135/100 ssu 43.9 35.1 20.4 API 05643 II 1-/ 1 T I T*BLE I SKIN PAINTING 5a?EF.I..:ENTS ON HIGH SOILING SAMPLES -- STEAIN CF r-ICS 1 ? ih AND a <o NUMBER CF applica API ,, SAMPLS tions OM z<n O P Ppi Q< number PER WEEK (n&) BIO L 0 .} I L/ n !j DA T A FINAL AVERAGE EFFEC MAXIMUM LATENT ORIGINAL TIVE INCIDENCE PERIOD RELATIVE NUMBER NUMBER OF FOR TUMOR CARCINOGENIC OF OF TUMORS INDUCTION POTENCY, MICE MICE !T.I./weeks) (5/^EL. in wks.) PMC 87 03:: 2 20 20 11 91/30 (23.815.0) (0.lQ+;fe) - 28* CFW 3 100 10 10 100/15 (10.81.6) o.w:j5 (20) (19) (84/15) 28 C3H 1 100 ' 13.(20) 13 (19) 85/33 (31+/51) (26.2+4.2) 0.31::^ 74 C3H p 100 14 (20) 12 (16) 100/4.7 (94/1+7) (35.85.3) -11-Io4 - 33* CFVT 3 100 20 18 95/15 10.3+1.3 *^1-o.22 33* C3H l 100 20 19 95/30 (19.4*3.4) o-sCis - 84 C3H 3 100 20 19 95/24 16.8+2.0 0.17+*^ -.02 850 90 C3H 3 100 20 17 100/38 (23.3*3.3) (o.i3!;gJ) 924 961 C3H 3 100 20 15 93/27 (17.713.D (o.i?!;") - 97 C3H 3 100 20 15 87/22 (17.9*2.2) (.i7!;J) 71+0 98 C3H 3 100 20 ll 73/39 (29.5+7.0) (o.n:;^) a 103 C3H 3 100 20 17 94/23 (15.9+1.8) (0.2:;^) The. pattern of response to the irritational effects of this oil was similar to that observed with dodecylbenzene, API 05644 I i! it I itU TnBLH I SKIN PAINTING EXPERIMENTS ON HIGH BOILING SAMPLES A NALYT C C AL DATA DISTILLATION |2mm. corr. to 760m PROCESS API SAMPLE NUMBER PRODUCT 750 GRAV <750* 925 >925 E, ITY VISCOSITY (*) (#) (50 w Delayed coking 41 V.'ax tailings, 1.2 131/240 9.6 49.8 .+0.6 50# benzene SSF Naphtha reforming M Polyforming 19 Cracked residuum 17.0 ' 58 ' Cracked residuum from kerosene feed 6.3 76 Cracked residuum 30.6 13 Cracked residuum 10.1 39.7/100 ssu 91 8.95 0 91 117/122 SSF 85 14.3 0 86 31/130 rj 91 8 ssu 86 1 154/100 ssu 63 15 22 8j f 37 Cracked residuum 69 Cracked residuum Hydroforming n 49 Cracked residuum 77 Cracked residuum 11.2 15.9 1 60.2/100 66 ssu 15 19 Sj 43/130 75.1 8.3 16.6 ssu 10.1 37.3/100 a,95 ssu 22.8 A'28/100 100 ssu 3.5 v9l o O o o 3 Cracked sidestream. 33.2 32.7/100 ssu tl 60 Cracked sidestream 24.9 40/100 100 ssu Distillation stopped when cracking occurred,, API 05645 Ti r^. N ' 3*P. ,Lp.) - TABLE I SKIT PAINTING EXPERIMENTS ON HIGH BOILING SAMPLES STP.A IN CF MICE AND rnp OF API applica- 1 1ITT v. NUM3EP PEP WEEK bio: 0 G I CAL DAI A crr3s 30M =h E-i < FINAL AVERAGE C M EFFEC- MAXIMUM <tOOJ tCC-4LU, Q *s! ORIGINAL NUMBER TIVE NUMBER INCIDENCE OF CF OF TUMORS LATENT PERIOD FOR TUMCR INDUCTION (mg.' MICE MICE (ToI/weeks) (5SSEL. in RELATIVE J CARCINOGENIC POTENCY, ; PMC ! 41 C3H 2 100 20 13 85/25 (23.1*2.0) 910 860 860 8901 362 6- 19* CF"' 3 100 10 (30) 0 CFW 3 100 20 (30) 76 CFV.' 3 100 10 (29) 13* CFW 3 100 20 (30) 13 CFW 2 100 30 37 CFW 3 100 20 (30) 69 CFW 3 100 10 (20) 49 C3H 1 100 20 77 CFW 3 100 10 (20) 3 C3H 1 100 20 7 (13) 18 (24) 9 (28) 86/37 (92/37) 89/30 (88/31) 100/30 (96/52) 13 (16) 28 13 (17) 10 (18) 92/29 m/38) 89/61 100/46 (94/46) 80/29 (78/30) 19 10 (19) O1 84Ai 100/38 (89/41) 0/62 (26.916.4) (0.071.02) : (22.4+4.0) 0.091.02 : 24.7*3.5 0.071.01 1 (25.6*3.3) (31 7+54) (25.4*5.1) <.07i;f) j .1:;2 | 0.071.02 1 (24.1*5.1) (o.o9!0^) ; (35.912.6) (29.3*4.8) 0.18+* ! .03 ; i(0.07!:2) - -: 01--1 + ,0 0 0 60 * CFW 3 i Number alive 100 after 10 (20) 8 (11+) 63 weeks. 63/48 (57/48) (43.4*11.1) API 05646 1 TABLE I SKIlf PAINT.ENG EXPERIMENTS ON HIGH BOILING SAMPLES A N A L Y T I CAL D A T A "--1 __ DISTILLATION (2mm, corr, to 760 PROCESS API SAMPLE NUMBER PRODUCT 750- GRAV- <750 925` >925` E.P ITT VISCOSITY (JO (JO (JO (F. 1 r*rw To Co Co tt 109 Cracked sidestream 7 Cracked residuum 9 Cracked residuum 24.1 46.1/100 ssu 20.2 34.5/100 oilOO ssu 18.U 37.1/100 ssu 97 2.33 700 ti 10 Cracked residuum 11.6 117/100 ssu 56.7 36.6 6.7 ft 31 Cracked 16.0 14.8/122 63 35.6 870 residuum SSF ft 36 Cracked 21.9 90/100 48 892 residuum ssu f! 40 Cracked 19 = 8 72.6/100 77.2 22.2 .65 300 : residuum ssu tf 71 Cracked residuum 22.8 59/100 ssu 77 21.5 1.5 sir ; 78 Cracked residuum 16.1 126.8/100 ssu 45.7 44.3 10 0 ft 82 Cracked 13.6 126/130 25.7 51.5 22.8 - re siduum ssu API 05647 E,P O F. 780 870 892 360 845 - 12 - TABLE I SKIN PAINTING EXPERIMENTS CM HIGH SOILING SAMPLES -- STRAIN OF MICE AND NUMBER OF API SAMPLE APPLICA TIONS wPPS-. S0H3 COQWc<O i<0ME4-i < NUMBER PER WEEK Eg.) BIO L 0 G I FINAL EFFEC ORIGINAL TIVE NUMBER NUMBER OF OF MICE MICE CAL DA T A AVERAGE MAXIMUM LATENT INCIDENCE PERIOD RELATIVE OF FOR TUMOR CARCINOGENIC TUMORS INDUCTION POTENCY, (T. I ./weeks) '5/&?L. in wksj Pmc 109* C3H 3 100 6 (20) 6 (15) 100/21 (93/25) 14.0+4.9 q pp+.20 Uo -.07 7 C3H 1 100 20 9 n/55 - - 9* C3H 3 9* CFW 3 10* C3H 1 100 100 100 10 CFW 1 100 31 C3H 1 100 36 C3H 1 100 40 C3H 1 100 71 C3H 1 100 71 C3H 1 20 71 C3H 1 5 78 C3H 1 100 82 C3H 1 100 10- 10 (20) 10 (20) 10 (20) 20 20 10 (20) 20 20 (33) 19 9 (19) 10 (20) 9 10 (16) 9 (17) 8 (10) 17 16 5 (9) 20 19 (32) 15 d2) 10 (20) 100/47 100/40 (100/4D 100/39 (94/39) 88/20 (90/31) 100/32 94/44 100/42 (100/42) 90/22 90/37 (91/52) o/9 89/19 (88/20) 90/33 (95/34) (36.2*4.5) 0.06+.01 (28.44.1) (24.6*7.0) (0.40:;^) (17.3*3.8) (22.0*2.8) 0.40!"?^ 0 XX (34.9*4.3) 0 2?+,^k 0 -.07 (34-1*7.4) (0.18+'|) (16.6*1.7) (27.1*4.8) (14.3*2.9) 0-55!;8| o-3i::Si - (24.16.4 i The pattern of response to the irritational effects of this oil wat similar to that observed with dodecylbenzene. API 05648 TABLE I SKIN PAINTING EXPERIMENTS ON HIGH BOILING SAMPLES ANALYTICAL D~A~ T A DISTILLATION (2mm. corr. to 76 PROCESS Po C a C * tl API SAMPLE NUMBER PRODUCT 4 Cracked sidestream 23 Cracked sidestream 750 GRAV <750) 925 ITY VISCOSITY (*) (*} 21.6 26.7 55.5/100 78.5 20.6 ssu ft 46 Cracked sidestream 23.6 56.7/1000 ssu 87.9 12.1 11 88 Cracked 22.5 78/100 69.3 30.2 sidestream SSU It 89 Cracked 27.0 51/100 100 sidestream ssu If 91 Cracked sidestream 20.4 57.8/100 80.1 17.9 ssu n 101 Cracked 21+.8 40.1/100 89.7 9.1 sidestream ssu 102 Cracked sidestream 23.5 53.1/1000 ssu 99 1 tl 104 Cracked 25.8 60.4/100 7.6 sidestream ssu API 05649 Y - 13 - 60mm; E.P i!.)l 838 TABLE I SKIN PAINTING EXPERIMENTS ON HIGH BOILING SAMPLES STRAIN OF MICE AND NUMBER OF applica- API SAMPLE flTMBEf TI0NS PER WEEK BIO L 0 G I CAL DA T A cl, ed CxJ <O OM COO CL, O< ORIGINAL NUMBER OF [mg.) MICE FINAL AVERAGE EFFEC- MAXIMUM LATENT TIVE INCIDENCE PERIOD RELATIVE NUMBER OF FOR TUMOR CARCINOGENIC OF TUMORS INDUCTION POTENCY, MICE (T.I./weeks) (5#?L. in wks. pMC 4 C3H 1 100 20 51 0/69 - - 23* CFW 3 100 20 20 100/19 12.51.3 n _| +.11 23* C3H 2 100 20_ 18 91+-/28 (18.5+2.9) 0 2^+o0^ Oo25-.06 780 46# CFW 3 100 20 15 100/24 (179-1.5) -u-:! 900 88 C3H 2 100 20 18 914-/26 (17.0+2.4) 714-5 39 C3H 3 100 30 28 89/21 (13.2+2.0) -28-:ok 830 91* C3H 2 100 20 19 95/23 (16.4+2.1) (0,28^'q^) a 790 101 C3H 3 100 20 6 100/12 (10.3.8) - 750 1028 C3H 2 20 6 6 100/27 22.6+3.3 (18) (16) (69/43) 1028 C3H 2 100 18 15 93/32 (23.8*3.4) -i9-:S6 101; 8 C3H 2 100 20 19 89/26 22.4+1.6 104 8 C3H 2 20 13 (20) 12 (12) 75/53 (75/53) (39.Oil0.1) 1 '-- , Number of animals alive after 63 weeks, The pattern of response to the irritational effects of this oil was similar to that observed with dodecylbenzene. API 05650 TAELE I SKIN PAINTING EXPERIMENTS ON HIGH SOILING SAMPLES ANALYTICAL DATA DISTILLATION (2mm, corr. to 760 PROCESS F. C, C. API SAMPLE NUMBER PRODUCT 8 Cracked residuum 750 GRAV <75C1 925' >925 E. ITY VISCOSITY (*) (%) (%) (OPc 11.1 110.3/1000 75 S3U 22 Dilution of No. 8 8-4 %0% oil No. 8, 50% (white oil + West Texas residu 8-16 50$ oil No. 8, $0% sec-amylbenzene 8-21 50^ oil No, 8, colorless fraction from No. 8 obtained by chromatography on silica gel__________ _ API 05651 r - 14 - .i 1 E-P. iduum) 5 T~~\ <M 1 00 1 TA 3LS I SKIN PAINTING EXPERIMENTS ON HIGH 30ILINS SAMPLES -- STRAIN pH1 MICE " AND NUMBER OF CxJ On s0 EHh <! WO ah <P API applica COO CHLi, SAMPI'E tfljVBER tions P < PER V.EEK (rr,g,,) 15 1 0 L 0 G I CAL DA r a FINAL AVERAGE EFFEC MAXIMUM LATENT original TIVE INCIDENCE PERIOD RELATIVE NUMBER NUMBER OF FOR TUMOR CARCINOGENIC OF OF TUMORS INDUCTION POTENCY, MICE MICE !T, Io /weeks) (5#EL. in wksj PMC 8 C3H 3 100 10 9 89/7 (5*. 4-i.o) 1"06-:lo 8 C3H 2 100 20 17 94/14 (8.5*2.4) (Oo77!^9) 8 CFW 2 100 19 17 100/14 7.5-1.8 >0.6 8 C3H 2 8 C3H 1 20 100 8 C3H 1 100 8* C3H 1 100 15 10 (20) 12 (18) 20 13 10 (16) 10 (16) 17 92/20 100/32 (100/32) 100/25 (100/31) 94/24 (16.6+1.8) (21.4-5.2) (0.47!;) (18.9-3.3) (o.w+!;lo) (15.9*2.7) 8 , C3H 1 8 C3H 1 20 20 13 (20) 40 12 (19) 32 100/50 (100/50) 97/42 (36.4*6.5) 0o20-!o8 (31.7-2.2) 0Uo2<33+-.o0052 8 C3H 1 50 40 3k 91/40 (27.5*2.2) 8 C3H 1 50 8 C3H 1 5 3.4 C3H 3 100 3-16 C3H 1 C3H 1 100 100 34 (20) 33 (40) 20 (30) 20 13 (20) 12 (18) 27 (33) 14 (20) 19 11 (H) 100/32 (100/32) 96/48 (86/48) 20.3*3.9 (35.0*3.1) IOO/23 (90/23) 100/52 (18.3*1.9) (34.4*3.9) 100/27 (20.9*3.0) (87/43)___ 0.20l;01 (0 T7+01\ lUol^".04; -22-!o7 API 05652 TABLE I SKIN PAINTING EXPERIMENTS ON HIGH BOILING SAMPLES A NALYT I : al DA TA DISTILLATION (2mm. corr. to ' PROCESS F. C. C. API SAMPLE NUMBER PRODUCT 12 Cracked residuum 750 GRAV <750* 925* >925 ITY VISCOSITY (*) {%) {*) 18.1 71/100 SSU ff 26 Cracked 7.3 344.2/100 61.3 34.1 4.6 residuum SSU H 42 ' Cracked 14.2 139.8/100 28.6 63.6 7.8 residuum SSU ft 43 Cracked 24.2 Pour Point 24.5 67.8 7.7 residuum 110 ft (44) Cracked 15.2 241/100 31.1 54.2 13.6 residuum SSU W 40 Cracked 6.0 91.7/100 47.1 37.3 15.6 re siduum SSU tt 68 Cracked 9.8 ' 81/130 43.3 44.9 11.8 residuum SSU II 73 Cracked 17.5 16.9/122 21 62 17 residuum SSF ft ' 85 Cracked 8.1 216/100 42.6 47.8 9.6 residuum SSU API 05653 _i TABLE I SKIN PAINTING EXPERIMENTS ON HIGH 30ILING SAMPLES API sampi 00% STRAIN OF MICE AN D number OF APPLICA TIONS PER 'VEEK DOSAGE PER w APPLICATION BIOLOGICAL DATA FINAL EFFEC ORIGINAL TIVE NUMBER NUMBER OF OF MICE MICE AVERAGE MAXIMUM LATENT INCIDENCE PERIOD OF FOR TUMOR TUMORS INDUCTION (T.I./weeks( (5#EL. In wks RELATIVE CARCINOGENIC POTENCY, PMC 12 CFW 1 100 18 17 88/34. (19o053) O0I4.6 +0 08 .24 1 12 CFtf 3 100 15 14 100/11; (10,,7o9) 0.42 + .26 11 26 C3H 1 100 20 19 100/20 (14.3*103) 0.65 + 0O9 ,08 42 C3H 1 100 20 18 94/34 (23.3207) (0.3&!0.0?;?) ,10J 43 C3H 1 100 20 15 IOO/37 (33oO2,,5) (44) C3H 1 100 20 16 94/40 (31.1*3.8) (0.25!;^) 48 C3H 1 100 20 13 100/36 (31.5*2.2) (0.22:;01) 68 C3H 1 100 20 l--1 O 73 C3H 1 73 85* C3H 1 100 100 100 10 (20) 20 20 15 7 (12) 19 18 100/17 100/22 (83/22) 100/31 100/25 (l4,,8i,,9) (062-!o5) (l8,,6lo9) (24.,,42,,5) (18011.6) o.kStfol 0o31_+ oOl(. .05 0.47+.06 r clipped. API 05654 T3TJT 16 TABLE I SKIN PAINTING EXPERIMENTS ON HIGH BOILING SAMPLES A N A L Y T I CAL D A T A DISTILLATION (2mm. corr. to 7 PROCESS API SAMPLE NUMBER PRODUCT GRAV 750 <750 9250 >92 5 ITY VISCOSITY (%) (*) (*) Houdry f! tt It <i ft 11 Cracked residuum 34 Cracked re siduum 50 Cracied residuum 51 Cracked residuum 63 Cracked residuum 4.3 71/100 5SU 84 16 0 28.0 47.5/ioo SSU 9 21.3 64/100 SSU 85 15 0 25.7 67/100 SSU 83 17 0 25.6 25.7/100 48.5 37.6 13.9 SSU tl 81 Cracked 22.7 40/210 40.8 52.6 6.6 residuum SSU Cycloverslon 22 Cracked residuum 25.6 84/100 49.5 44.2 6.3 SSU API 05655 - io TABLE I SKIN PAINTING EXPERIMENTS ON HIGH BOILING SAMPLES -- STRAIN OF MICE AND NUMBER OF API sample APPLICA TIONS flUM3ER PER 'VEEK 0 w y <03 OH <J COO Pli (31g) BIOL ORIGINAL NUMBER OF MICE 0 G I C A L DA T A FINAL AVERAGE EFFEC MAXIMUM LATENT TIVE INCIDENCE PERIOD RELATIVE NUMBER OF FOR TUMOR CARCINOGENIC OF TUMORS INDUCTION POTENCY, MICE (To Io /weeka) (5$SL. in wkSo| PMC 880 11* C3H 3 100 20 18 94/19 (15=012.2) <.22t;g) 615 3ll* CFW 3 100 20 820 50 CFW 3 100 20 an 51 CFW 3 100 20 19 84/32 (23=2*3=3) 0.09!;! 16 100/21 (15oOl,7) (o-is-lof) 18 83/19 (13=8*2.0) (.22!;|) 63* C3H 3 63 C3H 3 63 C3H 2 100 20 100 12 (19) 100 20 18 12 (15) 18 89/26 100/19 (100/19' 100/33 (20.3*1.9) (15.7*2=0) (2304*206) (0.141.02) 0 iq+04 UoXV~,03 0=17lf e 6 91 C3H 1 100 10 10 100/35 (25.9*4=9) (20) (20) (100/36) 31 C3H 1 - 100 3b 32 91/41 (30.7*2.2) 81 C3H 1 20 31 28 97/52 (37=7*3=1) 3 22 CFW 3 100 'is1 (3D .7 m? (10.8*3=1) -w:.L (20) (14) (100/17) ___ API 05656 17 TABLE I SKIN PAINTING EXPERIMENTS ON HIGH BOILING SAMPLES nArNALYTICAL DT DISTILLATION (2mm. corr. to 7^ PROCESS API SAMPLE NUMBER PRODUCT GRAV ITY C750` 9725500 >925j B. VISCOSITY (50 (50 (56) Solvent extraction 16 Phenol extraci 9.7 352/210` from Coastal SSU 900-X 62 n ti tt Distillation Dilution of No. 25 17 Duosol extract 7.1 2276/210 from Califor SSU nia waxy residuum, $0% in sec.-amyl- benzene 23.7 66.3 27 Phenol extract 11.0 173/100 80 from San SSU Joaquin naphthenic distillate 19 70 Nitro-benzene 15.7 99.14/210 extract from SSU Barbers Hill 60/80 . 85.3 14.2 75 SOa extract from TCC gas oil 14.4 38.3/100 100 SSU 25 700+ bottoms 24.3 115/100 49 45 from F.C.C. SSU heavy cycle gas oil 25-1 50 oil No.25 $0% (White oil + West Texas residuum) 21.1 320.6/122 SSU API 05657 f TABLE I I SKIN PAINTING EXPERIMENTS ON HIGH BOILING SAMPLES f-- " STRAIN OF MICE AND NUMBER OF applica API tions gAMFl^ in'NBEB. FER '.VEER' I C L 0 0 I c A L DA T A kJa,j som c--1J: <o-U> < 0o1 MJa, ORIGINAL Q a. NUMBER <S CF frag,) MICE FINAL EFFEC TIVE NUMBER C7 MICE AVERAGE MAXIMUM LATENT INCIDENCE PERIOD RELATIVE OF FOR TUMOR CARCINOGENIC TUMORS INDUCTION PCTF.'.'CSf, (T.I./v.eeks' (5/tFL. in wksj *V.c l6* CFW 3 100 10 6 67/73 (68.3*16.0) (C.03+,01) 16* C3H "3 100 10 7 43/70 (78.624.3) - 17* CFW 3 100 10 120) 8 (12) 87/62 (67/62) (35.4*16.7) (0.06-^) 27* CFW 3 100 10 (20) 9 (12) 89/17 (83/17) (11.4+3.0) 70 C3H 3 100 6 (20) 5 (lil) 100/42 (93/44) (3i.stn.5) <5 V ^ 75 CFW 3 100 20 13 77/51 (38.6i7.2) (0.05ft 01) 25* C3H 3 ' 100 30 in 95/19 (13.4*1.4) -+o03 25-1* C3H 3 100 30 26 92/40 (27.03.7) 0 0 " 0 +--,03^ API 05658 - 18 - TABLE I SKIN PAINTING EXPERIMENTS ON HIGH BOILING SAMPLES A N A L Y T I CAL D A T A^ I DISTILLATION (2mm, corr, to 76 SPECTRUM I API TYPE 750 SAMPLE OR GRAV <750 925 >92? % PROCESS NUMBER PRODUCT ITY VISCOSITY (%) (?) (?) ( i Straight run 1 Distillate 31.9 37/100 distillation # 11896 SSU n 56 Waxy Midcon 29.9 87/100 52.4 41.6 (2.0) tinent dis SSU tillate. Type .B ( .5) it 79 San Joaquin 16.6 63.9/210 rJ 13 86.2 naphthenic SSU distillate, Type C (2.2) 105 West Texas 28.9 54.3/100 72.9 23.4 3.3 paraffin SSU distillate. Type B (. 5) I tt Hk Type B (.63) 24.6 235/100 SSU M 115 Type C (.58) 23.2 234/100 SSU ft 5 Residuum from 12.8 572/122 26 26 48 I Wilmington SSP crude # 11885 (I ' See page 8 of Section A of report dated April 5? 193>2S for discuss^ of the classification of Straight Run Distillates by Spectrum Ty?9* API 05659 J) 830 855 TA5LE I si iy ?ai:'ti .iCII G b.'.:.?L~S a:.1 D NUMBER OF API applica sample tions (jijMBER prp '-.aZK : I 0 l c : : ; l la T A --------- r*- ?H 1 FINAL AVERAGE Er FEC- MAXIMUM LATENT Ota pfcm c< ORIGINAL TIVE NUMBER NUMBER OF OF INCIDENCE OF TUMORS PERIOD FOR TUMOR INDUCTION RELATIVE CARCINOGENIC POTENCY, (:ag,,) MICE MICE ( Tdo/week^ (5&TL. in wksj PMC 1 C3H 1 100 20 1 0/72 - - 56 CFW 6 100 20 56* cfv; 3 5fez C3H 3 100 10 (20) 100 ` 20 * 56 C3H 3 79* C3H 2 20 100 20 10 (20) 15 8 (17) 20 20 7 (14) 87/22 100/20 (88/22) 3/11 0/15 86/57 (71/57) (l5<>32o9) (15.4-3.2) - - rUi oJ--5^ + 0H 0O5 - -- (39.2+15.4) (o.i4+,?i) ^-.10' 105 s C3H 3 100 li+ (20) 13 (17) 85/33 (86/33) {23.2+4..4) 0.11 + *^ - .03 Ilk C3H 3 100 114 C3H 3 20 1:5 C3H 3 20 5 :3H 1 100 20 20 20 20 14 0/23 14 7/43 20 5/26 1 0 0/56 - - 1 Number of animals alive after 63 weeks0 Hair clipped,, The pattern of response to the irritational effects of this oil was similar to that ct. served with dodecylbenzene API 05660 - 19 - TABLE I SKIN PAINTING EXPERIMENTS ON HIGH BOILING SAMPLES A N A L Y T I C A L D A rp PROCESS API SAMPLE NUMBER SPECTRUM TYPE1 OR PRODUCT DISTILLATION (2mm, corr . to GRAV ITY 750 <750` 925 >925' VISCOSITY ($) ($) ($) Fractional distillation ft 110-1 in-4 Type 3 (.004) Type B (1.62) 61.0 .95/70f." 100 23.0 17/140 0P." fl 111-46 75$ No. 111-4 22.7 25$. Benzene n 111-5* Crude residuum 15.4 112/100pf {cl5$ Benzene) Dilution of No, 111-4 11 Fractional distillation 111-6 111-7 112-1 50% No. 110-1 50% No. 111-4 50$ No. 111-4 50% No. 112-1 Type B (.016) 65.9 ,896/70F.8 100 112-3 Type B (.043) 39-7 3.4/i4ofs Lubricating oil rt . i* 99 Type C (.5) 100 Type B (.3) 20.0 29.5 (475/100 l 52/210 rO 38 V ssu (307/100 < 53/210 V 0 1 ssu 100-1 0.2$ Antiox idant3 In NalOi 1 100-2 0.5$ Antiox idant, 2,6-di- tert. -butyl-4- 11 1 See page 8 of Section A of report dated April 5? 1952 for discussi<*j of the classification of Straight Rum Distillates by spectrum Kinematic viscosity (centistokes) 3 Sams type additive as that contained in API-99o API 05661 - 19 TABLE I SKIN PAINTING EXPERIMENTS ON HIGH BOILING SAMPLES STRAIN OF MICE AND NUMBER OF API APPLICA sample TIONS tfUMBER PER WEEK os g < uj CO O Q &h <Oh BIO L0 51 CAL DA T A ORIGINAL NUMBER OF MICE FINAL AVERAGE EFFEC MAXIMUM LATENT TIVE INCIDENCE PERIOD RELATIVE NUM3ER OF FOR TUMOR CARCINOGENIC OF TUMORS INDUCTION MICE !T do/weeks) ;5ELo in wkSoi POTENCY, PMC 110-1 C3H 3 20 20 191 0/55 tm iii-U *C3H 3 20 19 17 88/22 (17.6lo6) A 17+o02 081'-o03 111-4a C3H 3 5 20 20 0/3 - - iU-5* C3H 3 5 20 20 0/5 - - 111-6 *C3H 3 20 111-7 tC3H 3 20 112-1 C3H 3 20 13 (19) 7 (20) 20 112-33 C3H 3 20 20 99 C3H 3 100 100 C3H 3 100 100-1 C3H 3 100 Ik (20) 20 20 C3K 3 100 20 13 (19) 7 (17) 171 181 11 (16) 191 20 19 77/31 (58/3D 100/27 (76/14.1+) 0/58 (26o045) 22.43.5 = 0/582 - 91/36 (94/47) 0/80 0/3 (3i.o4.5) - 0/42 - 0o10i,,03 0 ii+^^- 8 -,,02 - - o.o7!;f mt - - rvi ooI -- s Number of animals alive after 36 weeks a Two papillcnas appeared at 20 weeks but regressed,, The pattern of response to the irritational effects of this oil was similar to that observed with dodecylbenzene,, API 05662 - 20 - TABLE I SKIN PAINTING EXPERIMENTS CN HIGH BOILING SAMPLES A > ALY TI C AL DA IA DI STILLATION (2mm. corr tc ?S( PROCESS API SAMPLE NUMBER PRODUCT GRAV ITY 750 <.750 925 >925* E VISCOSITY (%) (%) {%) ( Wax pressing n 32 Dewaxed paraf. fin distillate fraction from Lima crude 37 Dewaxed oil from No, 6 28.7 91.3/100 80 19.5 .5 ssu MEK - Benzol solvent dewaxing 1+3-1 Dewaxed oil from F.C.C, decanted oil No, 43 12.9 195/100 29.5 57 13.5 ssu Filtration dewaxing of No , 8 8-3 Filtrate oil, 11.3 85# of orig - inal ( ,94jT S) 65 21.4 13.4 Desulfuriza tion of 8-3 21 Hydrogenated 13.1 50.1/1000 74 heavy gas oil ssu by catalytic (.15* s) hydrogenation 20 6 Acid treating 62 Hydrolyzed acid sludge 14.2 7277/100 ssu 58 Effect of viscosity on ' tumor induction 111+ Straight run distillate, see page 18 of Appendix 24.0 235/100 ssu Retorting of oil shale 114-1 95% API-114, 5$ alkylpoly- 25.4 1756/100 ssu styrene 116 Crude shale oil 19.4 294.9/100 ssu API 05663 TABLE I SKIN PAINTINN EXPERIMENTS CN HIGH BOILING SAMPLES .CN :c 7An^ )25` E* j* % >) <? j " t *; CF VICE . *> *'* a: D NUMBER OF API applica:ample TIONS JrTMBEHj ?EF VEEK ! '; P* b BIOLOGICAL DATA eh Id S Ph O FINAL EFFEC- OCd <-1 <: 0 ORIGINAL NUMBER TIVE NUMBER COO .w_q OF OF Q Ph MICE MICE MAXIMUM INCIDENCE OF TUMORS (Tdo/wks,) AVERAGE LATENT PERIOD RELATIVE FOR TUMOR CARCINOGENIC INDUCTION POTENCY; !5#EL. in wks4 Pwr 32 CFV7 , . 1 100 (3 -6) 10 (20) 6 (15) 100/1+0 (87/1+1+) (31543) 5 $7* cpv: 6 100 20 17 3 0 5 43-1* C3H 1 100 10 ` 10 (20) (19) ,3-1 C3H 2 20 20 19 \l+ - 3-3* C3H 3 100 20 20 3 - 21 C3H 3 100 20 20 100/25 100/21+ (95/25) 0/5 (16,,9-2o0) (1952o2) - - 0ol+3-08 - 90/17 (11.2+1.7) (036-:ii) 100/13 ( 9.1+-101) (0,1+3*??) 0X 35c 62 C3H 3 Ilk C3E 3 11U C3H 3 100 20 100 10 (20) 20 20 9 (16) 111 11+ 33/79 (25/79) 7/1+3 0/26 (98o3-265) - - 114-1 C3H 3 114-1 C3E 3 20 100 20 20 15 13 33/1+6 0/28 - 116 C3H 3 100 30 20 25/192 - - 1 Ihree applications per week i or ^ek' thereafter, Panting discontinued after 13 weeks. six applications per API 05664 - 21 TABLE I SKIN PAINTIN'''- EXPERIMENTS ON HIGH BOILING SAMPLES ANALYTICAL D A T A DISTILLATION (2mm0corr. to 76q. PROCESS Fuel oil blending API SAMPLE NUMBER PRODUCT 113 Industrial fuel oil GRAV ITY 750 k750d 925 >92^1 VISCOSITY (%) (*>) (%) 8.3 30/122 SSF 43 37 116 jatalytically cracked resid uum component of API-119 119 10# API-118, 23.3 90# light cat alytic cycle cil (b,;+50 - 600) 35 -;j c ,, c id,; 38,8/100 SSU API 05665 TABLE I SKIN PAINTING EXPERIMENTS ON HIGH BOILING SAMPLES 1"-- API 5AKPL5 >f0M3EF 1 113 113 3 113 113 3 U3 STRAIN CP MICE AND _ Ch S0 WP* M h NUMBER OP <3 o S APPLICA O Oh TIONS PER WEEK 6ne.) C3H 3 20 C3H 3 20 C3F 3 20 C3H 2 20 C3H 1 20 BIO L 0 G I CAL DA T A PINAL AVERAGE EFFEC MAXIMUM LATENT ORIGINAL TIVE INCIDENCE PERIOD RELATIVE NUMBER NUMBER OF FOR TUMOR CARCINOGENIC CF OF TUMORS INDUCTION POTENCY, MICE M ICE (T,I./weeks) (5$EL. in 11k a,; PMC i 7 (27) 111 (27) 20 (27) 7 (27) 27 7 (22) 10 (16) 20 (25) 7 (22) 20 100/25 (77/27) 100/28 (83/28) 100/25 (100/25) 100/24 (91/40) 95/45 (18,8+3.8) (22.1*2.5) (17.5*3.4) (19.7*2.?) (18.1*1.6) (13.1+1.5) 22,5*.6 (26.6+3.5) (35.0*2.6) 0.16+.04 n -,+.06 0,1 -.03 0.16*.02 o.i9-+;J 3 U3 3 113 C3K C3H 3 3 3 113 C3K ** 100 5 1 27 (40) 26 (39) 20 27 (36) 26 (38) 18 100/23 (100/25) 100/26 (92/34) 78/54 (16.3*1.3) (17.5*1.3) (16.9*1.5) (13.9+2.1) ''v'46.1 0.17*.02 vo. 04 3 113 C3H 3 4 113 C3H 3 20 20 20 20 16 100/2.7 (19.2*2,1) noi177^-*o04 20 10C/26 (18.3*2,1) 0 ig+ 03 118 C3H 100 f 2/m onth) 24 (30) 21 (24) ICO/50 (100/50) (3a.9*4.3) n 1, a+02 04b=.17 119 C3H 100 ( 2/mor.rh) 119 C3H 100 (2/week) 29 22 (27) 25 19 (23) 40/54 95/27 (96/30) - 21.6*1,5 - 0Uol8io+,,#,o0^i r-- Approximate age of mice at start of experimei Approximate age of mice at start of experimei Approximate age of mice at start of experime] Approximate age of mice at start of experimei 27 weeks, 23 weeks, 20 weeks, 14 weeks. API 05666 TABLE I SKIN PAINTING EXPERIMENTS ON HIGH BOILING SAMPLES PROCESS AFI SAMPLE NUMBER PRODUCT [Fractional distillation d-61 0 - 93/fc fraction from API-Q 8-7* 9.3 - 19.456 fraction from API-3 8-8* IQ.I4. - 30. 3>% fraction from API-8 8-91 30.3 - 39.8 fraction from API-8 ESTIIvATSn CGRhLCIto oOILLjg Ha i IC;V F. at *35 - 3:0 rt-101 3908 - l4.9<>656 fraction from API-9 3-111 496 - $9.2% fraction from API-8 8-121 59.2 - 69.15t fraction from API-8 Diels-Alder reaction Fractional distillation 8-12al non-adduct from reaction (95.45= of 8-12) 3-131 69.1 - 73.5/6 fraction from API-8 8-114* 78.5 - 80.8^ fraction from API-8 8-151 Residue; 30.8 - 100^ 3-18 Proportionate reblend of distillation fractions, 3-6 through 8-15 50- solution in benzene. 33,,3J? solution in benzene. API 05667 T ~ 22 ~ TABLE I I SKIN PAINTING EXPERIMENTS ON HIGH 30ILING SAMPLES BIO L 0 G I CAL DA T A ED ED O' Cl :U 0 5 STRAIN OF MICE AND NUMBER CF API APPLICAsample TIC NS HUMBER PER WEEK 0PaL5. 5OH C-'j oO t<oo ^ P- (mg.) FINAL AVERAGE EFFEC- MAXIMUM LATENT ORIGINAL TIVE INCIDENCE PERIOD RELATIVE NUMBER NUMBER OF OF OF TUMORS FOR TUMOR CARCINOGENIC INDUCTION POTENCY, MICE MICE [To I o /weeks) (5#F,,Loin wkq Pmc 8-6 C3H 2 100 20 141 0/66 - a 30 695 710 8-7 C3H 2 9-8 C3H 2 8-9* C3H 2 100 100 100 20 20 20 1 16 lb1 18 0/74 0/74 56/62 (58.8+13.4) - 7*5 8-10* C3H 2 100 20 16 94/44 31.2*3.4 300 3-11# C3H 2 100 20 20 90/33 (24.62.5) 0.15!;^ 830 8-12* C3H 2 100 20 14 93/28 (19.5=3.1) -23;;g| -5C - < 3 *17 sCs 8-12a* C3H 2 100 9-13* C3H 2 100 3-11+* C3H 2 100 3-15* C3H 2 100 16 20 20 20 16 100/26 (17.6*2.1) 14 79/25 (22.1+2.5) (0.20^|) 12 25/21 (26.1*4.7) - 13 92/35 (28o0+2o6) 3-18 C3H 1 100 10 (15) 10 (14) 90/18 (37/39) 13o3^2o5 0 72+0^^ Uo' 16 1 P-- -- Number of animals alive after 1+5 weeks API 05668 - 23 - TABLE I SKIN PAINTING EXPERIMENTS ON HIGH BOILING SAMPLES PROCESS Diels-Alder reaction #1 ft It ft fl tl API SAMPLE NUMBER PRODUCT 8-23a Crystalline material from 1st curomatographic frac tion of Class A1 adduct regenerate, .34/2 in benze o-23b non-crystalline residues from 1st and 2nd chromat graphic fraction, 1.5% in benzene 3-23c Crystalline material from 2nd chromatographic fra tion of Class A1 adduct regenerate, .34/6 in benze 8-23d 3rd chromatographic fraction of Class A1 adduct regenerate, 0.8 $6 in benzene 8-23e 4th chromatographic fraction of Class A1 adduct regenerate, .57% in benzene .i 8-23f 5th chromatographic fraction of Class A adduct regenerate, .51% in benzene 3-23g Class B1 adduct regenerate, 1.5% in benzene i 8-23h Total Class A adduct regenerate. 2.2% in benzene Solvent extrao tion with cone, HaS0* It 8-19 8-20 Extract from API 8-10 and 8-11, $0% benzene Raffinate from API 8-10 and 8-11, 50% benzene t! 23-1 Extract from API-23, 2.0% benzene ft 23-2 Raffinate from API-23, 20% benzene Fractionation of API-12 12-7 Non-adduct of Diels-Alder reaction on raffinate f sulfuric extraction of aromatic fraction b.130-13 at .2 mm. 680-800F./760 mm.), benzene Class A aiduct was tnat normally soluble in $% aqueous NaOHj Class adduct was not soluule in either 5% aqueous NaOH or benzene sf-er hydrolysis. API 05669 r r> --_ -'rao. azene aatofracazene t r-'-'t t zent te from D-l80eC TA5LE I SKIN PAINTING EXPERIMENTS ON HIGH BOILING SAMPLES STRAIN OF MICE AND NoMI EE OF API applica 5.4MPL2 tions ^MBER PER WEEK BIO L 0 G I r023h5 COwh <? FINAL EMO<J C MO JC0o ORIGINAL NUMBER EFFEC TIVE NUMBER Q < OF OF (mg.) MICE MICE CAL DA T A AVERAGE MAXIMUM LATENT INCIDENCE PERIOD OF FOR TUMOR TUMORS INDUCTION (T-I0/weeks] (5&EL. in wks' RELATIVE CARCINOGENIC POTENCY, Pmh 5.23a C3H 3 15 20 17 0/54 - 3-23b C3H 3 15 20 11 100/51 AJ 38.1 rJ 0o05 3-23c C3H 3 12 10 . 6 o/543 8.23d C3H 3 8-23e C3H 3 8-23f C3H 3 15 14 (20) 15 9 15 20 13 (18) 8 100/36 (100/38) 13/484 16 o/544 2904-207 - 0.08.01 > = - S-23g C3H 3 15 14 13 92/50 43054.2 o.oi+t.oi 3-23h C3H 3 15 13 12 100/1+3 ' 35*213,4) -7i:o2 3-19 C3H 2 100 9 9 22/32x - - AVM)8-20 C3H 2 100 6 (9) 6 (9) (22n850) Ooie+o^ ".05 3-1* C3H 2 100 20 19 90/26 (18.213 a) 0 28+ '02 -.09 23-2* C3H 2 100 20 19 100/44 * (25.7*3.1) n1i6+-,t0^k *2-7 C3H 2 20 20 18 33/465 - - -------, Minting discontinued after 2o weeks,; 8 30 weeks; 3 37 weeks; 32 Weeks; 5 ^3 weeks. API 05670 - 2k - TABLE I SKIN PAINTING EXPERIMENTS ON HIGH BOILING SAMPLES PPCCESS API SAMPLE NUMBER ______________________ PRODUCT______________________ Air oxidation (cobalt naphthenate catalyst) 83 83-1 Two hour oxidation product Four hour oxidation product Chromatography 3-5 Aromatics of No. 8 from Silica Gel ft 8 .1 ,, 8-13a First aromatic fraction, 2% , of API 3-13 If d-13b Second aromatic fraction, 2.4#1* of API 3-13 If 8-13c Third aromatic fraction, 8.7$*, of API 8-13 8-13d Fourth aromatic fraction, 56$1, of API 8-13 tl 8-l3e Fifth aromatic fraction, 5.6#1, of API 8-13 It 8-13f Sixth aromatic fraction, 2$*, of API 8-13 tt 8-13g Seventh aromatic fraction, 16.9/6 , of API 8-13 It 8-13h Eighth aromatic fraction, 2.4$ , of API 8-13 ? 8-131 Ninth aromatic fraction, 6.9$ , of API 0-13 M 8-17 Reblend of chromatography fractions of API-8 t! 12-2 Non-aromatics (cut with 20$ heptane) ft 71-1 See page 34 of Section A--i+ of report dated April 1952*for description tt 71-2 Proportionate reDlend of all fractions from cnromatograpny of aFI-71 It Non-aromatics of No- 3 from alumina 1 Tested at this percentage by weight in benzene. Proceeding colorless non-aromatic fraction, 43$ of API-8-x3, proved to bo a non-accelerating solvent when diluted 50$ with benzene. See API-266 on page 37. API 05671 - 21+ - TABLE I SKIN PAINTING EXPERIMENTS ON HIGH BOILING SAMPLES -- STRAIN OF MICE AND number OF tfl applica- 5A.MPI-e kjmssr tio: IS PSR WEEK BIO L 0 G I CAL DA T A r3' CLi 'O a-1- Ehh tq <0* FINAL EFFEC- MAXIMUM AVERAGE LATENT C<TO- ^CA** ORIGINAL TIVE NUMBER NUMBER n < OF OF INCIDENCE OF TUMORS PERIOD FOR TUMOR INDUCTION RELATIVE CARCINOGENIC POTENCY, (mg.) MICE MICE (T.I./weeks) [% EL. in wks] PMC 33 C3H 2 100 20 14 100/17 (12.7+1.7) (o42i:8|) 33-1 C3H 2 100 20 16 94/17 3-5 3-5 3.13a 3-136 3-13c C3H CFW C3H C3H C3H 1 T 3 3 3 100 100 20 20 20 20 ' 20 15 15 15 12 19 12 12 13 92/22 79/28 0/30 0/30 69/30 3.13d C3K 3 20 15 8 87/29 8-13 C3H 3 20 15 7 43/29 8-13f C3H 3 20 13 12 25/25 3-138 C3I-: 3 20 15 9 100/27 d-l3h C3H 3 20 15 12 92/29 3-l3i C3H 3 20 15 10 70/29 3-17 C3H 1 100 10 (20) 8 (17) 100/17 (94/30) 12-2 c?w 3 100 20 151 0/83* 71-1 C3H 1 20 30 30 83/64 '1-1 C3H 1 100 15 14 100/16 71-2 C3H 1 20 30 |j`<4 C3H 3 50 30 t 23 22 93/59 o/4o j Number of animals alive after 33 weeks. Painting discontinued after i+1 weeks. (12.4+1.6) <o.w!;g) (I8.7il.8) (21.2+5.6) - ~ 27.4 24.1 - (21.02.8) - 24.7 ^ 25.3 (12.4*2.3) o.44!"J 0.261-J1 .07 v 0.09 ^0.10 (o.iig;0^) 0.10 /U 0.10 -8o-:i8 (39.0*5.6) (11.11.5) (34.6m*5.0) 0.18!;^ 0.22+?3 _".0o API 05672 - 2$ TABLE I SKIN PAINTIN'} EXPERIMENTS ON HIGH BOILING SAMPLES PROCESS API SAMPLE NUMBER PRODUCT Distillation 1 plate vacuum) IZ-k 12-5 Aromatics b. 510-7i+0F. (corrected) Aromatics b.*^900-1000p. ( g,,/100 ml. cenzene) 12-o Aro.matics residue* b,y 1000F. go/100 mi. Denz* API 05673 T - 1 TABLE I ^I ski:; painting on his:-: soiling samples ^-- _L CLOG I C A L D A T A tj* DOSAGE PER" A P P L IC A T IO N s.^ iTR.i lit API _ 5.; ?L- "l2-4* OF MICE ML ITUi-.__L.rC CF ,.p?li.:.v' tf ; s per ..ask U X - 100 FINAL EFFEC ORIGINAL TIVE NUMBER NUMBER CF MICE CF j ' T *1 10 (20) 10 (16) . '.AAIKUM AVERAGE LATENT IRCIDEACE PERIOD RELATIVE OF FOR TUMOR CARCINOGENIC TUMORS INDUCTION POTENCY? (To I,/weeks) (5/SFoLoin wk$ Pmc 60/34 (56/31+) (27.2+13.2) 12-5 cfv: 3 100 10 (20) 12-6 CF''.' 3 100 10 - (20) 6 (12) 8 (12) 100/34 (92/34) 75/39 (67/33) (23.6i58) 0o07-!oi (32.2-120l) U#U--o01 API 05674 CONCENTRATION STANDARDS WITH METHYLCHOLAN'L'HRENE IN BENZENE Area co ve re d by 0 .1 yram Benzene. Number o f a n im a ls a liv e a f t e r 36 weolcs API 05675 co KE<aHo-i ECO-i C<Ed- O 1CAM s\O \AOl' Cr--Mt( txHOif - 27 - UCM\ 1CAM x\COM \CCOM \O' \oH O' \ \1CoAM HrH9 tsriDH-9 +-CrrrdHHHO1t" OrH' %irHH# o < CM +-rAd1-- IA \CM \oH 1CAA \ CHA \ CM O O' \ >o rn 1A eg rH iH +1 c xO CrHO cx- 1A rH MCM CHM rCHM xO \o o > O H \oO o o o o H rC~M so o CM \oO OCM OO rH oCM OCM COM oCM O' oCM COM COM COM OrH orH CoM CcM C3M TABLE I I A rea covered by 0.1 gram Benzene 1OCl--AAi 1O(HAA frH^\ O xCOrOHM 1UCOMA\ s1OOAJ. 1COCMA- --r. 3OO S5 O PJOcog CA . CO CA CA M H MHgS <! S Cscajo K 5CSfcHK a a S3 Ka Si 3 CH E-l W CA CA CA CA CA CO O < Pi o U O CJ O OCM O CM OCM O O oo a * ao w M MO O Eh o a <H o a\ O E-I rH rrH o rH erH O rH r- 1A 00 rH CA xO O oo "CoM -d oCM o CM 1A X) OO CM CM CM CM CM a M- a CO ro CO o CJ o OCM o CM O O *H CO CM xO CO % CM CM ooO CO CO 1A xO O CM CM CM API 05676 API 05677 A re a c o v e re d by .1 ',i*ajn J e n z e n e "I'.J ^ N T h A '' .lON A T /U ii A.cDt) W IT ii MU Jr *ifYi.C!I L A l.T k iii'.iiE I N B K N ZE i; API 05678 ' :\J nI r e a d n b;r .i , I ' iu ifils 'iV . benzene. ! ": ?i ( - 30 - r^CNj o *o* tr! Vi 1OA o HCM o*o+1 XI o f\J H O O* V I I aam. CM O CO CM 1A t iH CO CM CM VI VI +1 VI CO vO CT' CM o c\ <r\ c"\ -d" CM CM W SO nX 5 ?B OT aw (do H Q ft, O fc m o as\ <0 D aa Sm H EH O VN \ P>H vo3 ^d^d- om 00 03 nO -dN CiHO N o o H < EH W Sa o a fc u cn OvO vO HSOH CM iH r-1 cufeB a OO H CM rO\ OO O H CM CM O' o o H CM BefOeHiJOH CH m o~\ <n cn MW < fe O Eh Ha CO a*a O &oH O pX) 0p3 -d- -d P- oo wa IH p w -d" P~\ Ph Ph g CM CM CM CM cn CM CM CM CM CM CM CM CM co a API 05679 Dosage p e r a p p lic a t io n was 100 mg d e s c r ip tio ni EXPERIM ENTS IN V O L V IN G THE A P P L IC A T IO N OP S O LU TIO N S OP METHYLCHOLANTHHENE IN V A R IO U S SO LVENTS W1 >0 >-1 h la 0 0 EhHHZ C <uuzy s W <OD OS O Ph os a ^ H 0O M O&hQSM JaJ < M os Q ,, C P Ph O sVi. C pL| M LTN H W ' SO m D 3 CO ii S Ctj OS.S M Q 0\ XMOS <5 0 p M SB M E-i Eh 1 CS tJ O Ct| EJ W < a > ,c h u B EE H s 0 H H fc Eh B S aE b rH 0 +1 s 0 H +1 Cs nx CM W O <^\ \ O O rH cH j <K BM M H Q fe 0 C 50 H MP S CS .O W 1B ' C5 OS M O < y j h to CO Ph Ph h E O Ph Cw 0 <0 BW & a MO -n rA < h 0 fi 3 3 n PLS 2 3aMMgQpSs O P,;l< w B << Ph (aUSAIOS 3 00T/*S) IOIIVHJ.N30KOD 0 CM O O rH c*n K O 0 r^i 0 EH CM UN 1 ' >M J Ph 0 <! CO (Tb 0H S< 505 0 dj '" 1 B H S3 Eh Eh OS <3 0< EH CO M hj a Ph Ph CQ <as 3p 0 S O UN \ PN \ CM CO fH CM - 31 - r'NrH OO f 1 CO 0 0 f*NCM OO f 1 O CM O 0 2 0 ;. r-YM OO +1 H CM OO 4- | CO H O H 0 tl cn 0 0 CO CM -d" CM rr\ H H +1 +1 4-1 +1 sO H O fH- H UN UN C CM H H jH 4| Q -d UN Oh CM H CM CM CM CM \\ \ CM \ CM \ OH O O O 0p O O O HH O O \ C\J sO OVD 0 H CM co H O CM *N 0 0 O 0 O CO H CM CM CM CM rr\ OOOO O CM CM CM H O O H m PN 0 O0 UD 0 O0 ro C\J 00 CO H 1 UN CM IH HH Ph <H Ph < PN c*N PN O0 OH O0 C\J (V 00 c*n H rH 11 HH Ph Ph << OOOO Saas CM CM CM O UN UN UN UN \ . \ \ \ CM H H CM H H CM \ON \ H \ -d* CO sD sO CO H -P- -B- -dCM CM CM CM 112 110 281 r-N M-* -d" 0 0 0 0 TO n C n CP 0 S \ITS tM 20-M ethylcholanthrene W ilm in g to n Residuum ; (M G ). see page 10 o f Table I fo r H (\J m See page 19 o f T a b le f o r d e s c r ip t io n . API 05680 weeks CONTROL EXTERITJENTS W ITH VARIOUS SOLVENTS BER P IC A- >NS4 WEEK 32 u 55 rH u\ r--1 XA O xa \\\ CA XA \ O XA % O O rH XA XA XA \ \XT\ a- CO XA -sj" CO CO CO \a\ CO \ XA A- CM \ CM H \ r- \ CM CM CM \ rH rH r- \a * m a ^^ 0 so 3 'CO O M CO S3 fe P O M\ ah 0a u u os +5 c s <0 H OO CO CO CM r- C-- XA \ \\ 0 OO H -drH \ O rH p~ \ O O CM CD H nO N \\ OO0 0 \ 0 0 a a ra so a S3 s Sa3 CO a O H* O JH <0 fc O OS Sa \ S S3 M E-< H Cr--O CC--O t"CA \\\ 0 0 CA CA CA -d" \rH \XC--A O rH rH 0 CM CO s v\O S 0O 0 .0 \0 M >K p< h &h a CQ (i, a O BOSOH hasa)5ss s a tH CM vO vO CM CM CO rH p <K sa a H C ft, O *a0 *s-J p--1 O CO 0. CO CA CM Sm<3 !poa :5 O M E-i S CO a CO 0 0 r0Mo O 0 CA C0O rH CM H CM CA CO NO O CM CM rH rH CA rH O 0 0 O r>- CO CM CM -d" rH ! 0 W C0O SS P=H C3 W C0O 0 Number o f a n im a ls a liv e a f t e r 36 w eeks. Number o f anim als a liv e a fte r 33 w eeks. Age o f anim als a t s ta r t o f e xp e rim e n t, Dosage p e r a p p lic a tio n was 100 mg. cO ca CA CO ro <A CA CA CA ss <&U. h aan l S25) SP> O CO rH H rHH 1C O O H NO P rl P H OC 'O P O 1 rtH c N et 'O p 0 rtH c 3s m 1a p c N c O 13 Cl 1H--1 Ch H rH Pp PP Na p n p pd Pra t 5 * O p CO a a Q O mE3| <p5 pSi a55 :)( 0 XA rH CO CM CO rH CM -J CO CO rH CM CM CM CM CM CM CM CM CM CM CO !Z r--I CM API 05681 EXPERIMENTS IN VC rVJM A Tr" ' A P P LIC A T IO N OF SOLNTIONS CF SYNTHETIC CAFC IMOGENS IH DOEEC fLLENZEM E o M COG >y hoo Eh o z c; <ay s J M 9h ^ yoj U Qa O 20 haa s weoo cr 3 s h c <0 ft 5 Eh ft Eh O W |H & >2K3J <2 tiq O z Eh Ift H ii i| p J1 \J rH on +1 o o cH CA r- t"'.^r o O- -H oo +1 + 1 f ! - 1 4 1 o bN 4* rr^ CM IT. CA r*\ oO O o H rH H 1 +1 CM IA rH o -H +i CA CA. -d- o p- ON o c- -d" rH AJ CM + +1 4-1 H +1 o 'lA .A O' CA -d- o H o o CA rH H rH rH * W gg r; P ft CO s S K'v H O ft O ri o a h <J O D SZ Eh Eh M ft. (K-H a- --T* -d- CO O fO u\ co-d CA CO C- '"a 'wA -d rH M CM H N \ ` \ Vn \N \ s \ CO o iH COO sOlA o OH c32 o o O o o c o o '-r H rH rH rH rH 'W' tJ 1 < O W ft ft z ft > CQ ft O H fc H S O h ft ft EH p E ft a ft <2 aft ft CQ ft O C sOH M5 3 es a o CO rH o rH CO CM rH co-d- 1A IA rH rH r~ "W o CM o CA o o o o o o o o O CO O O CM CO CM H OJ H CM CM CM CM CO W< C5 O Z- < K ft O OoOo O OOo Oo to ft A r-i ft o o O o o o o o O o C ft ft j? H r--i rH rH M H rH rH rH rH <2 ^ 1 ft CC < ft ft owy 02 ft H ? 5 O ft O 5 ft Ec ft Eh ft <2 ft CA C^i <r. . r^, CA r*. CA a M ft <2 ft O a a 2 a a & aa COM fO CO ft CA pi-t rn ft CA EH 3 ooOO o o C2 u o C2 CO ia a o ft ft H o sO JD rH rH 22 -d" -d* O' U Eh O o O rH rH -d- rH rH -d" a<h o o 6oO o o O --I rH rH O Eh oo o oOo o o ft ft ft! S %* ro * A Ph fL, P3 i-W vO vO in CA rH rH CO <iaa CM CM CM CM CM CM rH rH rH o <a CM 0J CM CVJ CM CM CM CM CM CM co a fTt3 U fOt A o m as o A fGt oH A ft * Ctf U C\l P MI ft P < G P P. I--I >> V Q, XO h ft ma! o 12 +5 !u -G P > cH G M G G as G > A |ft -p o faGtS G o o ft A o fdPtj F> O ft -I ft a s <n c 20 i>H a * C>C ibdfN- rCH * b-, p o 0w fet C tr G x ft m Ha) c S n nas ^222>> P J aft c..: a! SS d -P c M ft 0, aGC 0 -H -P C H- ft h rt o S>-P G 0.22^ O C H N04JH Oc wr Ci O +5 >J r ffl 22 O H <h C O I 0 as 0 -d"d -P w rX CA o Cl a O ft *> Eh ft < rH CM C^ -C API 05682 T w oM 2 fc O 2M W a a h o e-i EA.FL'GIMGNTS IN V O L V IU t: '!'lUi APPLICATION OF SOLUYLC.io OF CARCINOGENS H i DCDECYLBEUZENE' NUMBER DOSAGE OF PER API F IN A L EFFEC ORIGINAL TIV E NUMBER NUMBER OF OF MICE MICE MAXIMUM AVERAGE INCIDENCE LATENT PERIOD RELATIVE CONCEN SAMPLE TRATION NUMBER [g ./lO O g .) STRAIN OF MICE APPLICA APPLICA OF FOR TUMOR CARCINOGENIC TIONS TION TUMORS INDUCTION POTENCY, PER WEEK {m g . ) (T .1 ./w k s .) ( 5 $ F . L . in w k s .) PMC - 3U - O"0hn 3 ou hoo" + 1+1 -d cm PA -d oo v\ co ho +1 ua P.A o h -o >-_d o pa hoo*doo +1+1+1 o r- rP.A .rH .rH o oo _d cm rH i--t +-d O +1 OrH'* --* h f\J +(V1J rH o..p- .fCM PV CVJ P+A1 O+1 +-d rH f**- O rH rH rH 1 0 0 /3 1 100/19 1 0 0 /lk (1 0 0 /1 8 ) 1 0 0 /1 2 (1 0 0 /1 2 ) 89/U i CO rH CVO rH rH OO rH rH C*HO fr\ ^ rH O (AO OO O tf\ C UA OJ HCJ rH C\J CM H HH 100 20 5 100 20 100 r"\ pa pa pa pa pa aa POA POA a PUA a aa POA POA POA Pc.- Cr--O 3d- Cr~VJ- CrM- Cr--O H rH PA i--i rH . rH. oo o o oo O 0s -d -d CVJ CJ rH rH rH CVJ XT' tf\ \A H _d j -d CVJ CVJ CJ CVJ 1 0 0 /2 3 (1 0 0 /2 3 ) C8 O Jo=I H XS * UV -cMdoi Px > P a pH O i--t sPii'O p s En 3 3 c to au O ! 5 5* oC Pi a , UI J3-dn ot E-i a CM 05683 d u ct obtained by a lk y la tio n o f benzene w ith p ro p yle n e te tra m e r (alum inum c h lo rid e c a ta ly s t) E X P E R IM E N TS ON A C C E LE R A TIO N OP C A R C IN O G E N E S IS BY S P E C IF IC SO LVEN TS -*rtor a o 3o HASH H20HDtiOH d H E<h 0cq3 a (JAhO PQ pCl, 6h Mtr\ - 35 - LA vO CO O' +V ftxt A 4-1 41 CM O (V AJ M 00 fl 4-1 41 4i O' O CA AJ 0 CO CM sO xt- -d CA M H CM H sO CO O 65 so tAoX P s co S 65 05 Q H Q ft OS X H O s\ <3 U X s3 H Eh M 1# Eh I 05 4 a m cq ,, 65 s! 65 > CQ (Z4 O ZfcHSOH H&.HP S3 few A- O M A- H CA \ -d-x xt \ AJ \ CA oo O O CA o o O O rH rH rH rH AJ Al AJ AJ OO OO HH OvO LA vO LA AJ C\J C\J \ \\\ o oO o O O' o o o o H " H rH rH H CO OvO O' O MH CM vO a a CO o O MH CA A O oo CO H AJ O A-O O O O O AJ Al AJ AJ AJ CA C0 CA CA CA <A CA CA 'O vO CA CA *3 HW <60 05 O M Eh S C0 03 e CA OO 03 03 03 CA P"\ <**\ OO O 05 r-\ to S 03 05 &4 CA CA OoOO 65 w CO CO O' O' O' O' O' A- o O CQ 1 i1 i 1 LA LA AJ AJ H gS 3 65 S Q P H a < MMM Oh a Oh < <5 M Oh <5 1 111 MMMM Ph Ph Oh Oh 6>5 O XOh <5 J c < 65 H 65 CO 65 M WOE 65 Eh Oh CQ X 05 65 M6, 05 HKEhH (LMog < Oh 65 S XSP C5 3 C\J O' CQ 1 PH P Oh < :Jc H O' AJ AJ O' 1 CQ HP Oh P < a p p :|s O' O' O' 1 1 rH rH AJ AJ AJ Al LA AJ CO A- o p 1A CQ AJ CQ p 1P 1P Oh M P M P Oh Ph << O' 1 CA VA AJ AU\ * ALA 1 rH AJ AJ CO O Al cAooJ I rH AAJ X2 +> T3 C H ci X JO o 05 pP m a H <H (D rH d O o ao XdI -oo as i 0 rH 1 AJ X! -H X rH Xx! S h -d d <H d as -P dh Xo> x-- P Ah P O Ph do d X E d boO d H o u aS o XI X M d H -P d M ft 2 M -H E >>,d x i; O *H O Hd d o M *H >5 Xx d dO aO _d Xai xMax dm o p n xa s d d cm as d t3 O dn o xs S -d d at CQ Ph f o a rH C. x o -d C- d h xs H < X Ph X n h a d m a c H>O M>Oai X EH E g XEh a A TJ Sh -H o O a EH CO P P CO A CA . API 05684 EXPERIM ENTS ON A C C E LE R A TIO N OF C AR C IN O G EN ESIS BY S P E C IF IC SOLVENTS m asx MCPOSOHO S < Z o fO El C aCDEchJM05Eh5O3^H < J a O Z6j fa H-feS. U\ C\J +1 CO CM H - 36 - tA CM +ci CO -d -d- vO O CM O' o 1--1 CO sO CM CO CM A- r~- \\ \ \\ \ CM -d o A H P- H O' tA CO co sO o vO 4n vO nN SO vO CO H CO CO CM H O O in IA O -d- -d- * -d- -d- ro oo CO CO I ro PO co CO CO cO z H Cxi < fa o 05 O H Ei 2 Z CO a CO a CO a CO B co a CO a CO a CO o C3 C3 53 o C3 C3 a H H OO rH a i--i QH 6a Qo a oP CO l S3 a z Q o Q d 43 H H P1* < -d 1A CO O' % O' CM -d- CM CM CO CM 1 i | ww Haa a< < <5 -d CO | M a < co i H a < O' _d -d cm as co CM 1 1 1 HH h aa a <} <5 ! CO CQ H H gH H 55 Z QHH O*Z . n cd s m aMW2 <asg S* o H CVJ nO 0s o CM CO CO CO CO co CO _d CM CM CM CM CM CM CM A -P pH > a c H pas o a h o g c a ftp o oc Q pa M C H (>. C EE h pa ft d i H d H Q) <D (D * H C (U O O P O as C (t oa u p d s a oah uap e d i tn oH po 1 ft o Ai a h HP H U fl a * oh p ft na * a c o H c bO O C H O u as V c HH Cddd p > sj na ua COOP a p H * PC bO a--, c d c ah n H fot Qa a, c P *c '-- ft nh c I *i d O Jftl ft PCOOH SH d O* I a] tc H +5 H Xh'OO F>>a HaAP COH 'JD -P E u H c O O d H 0) H >>-0 e C -P >> C H O ft ft n a) o a) ao i p * Z ftH ft O 1 HH H >p C CM O C h d o H ft O' O-lfl H at as P 43 u a g d aoa Zn0 o OSH HC C1O4 ft P UN oo H C ft d E hc ft _ao pH p Hft fOt Bft H 3 ft ^ ft O ft "85 HP>HOfHt O H A Edh O C O 5 ma3l O E ft as C ft te O ft tO 0 h A ao o o u d h H o Hm facamn g OP co z o as O O cm co^d-iAivO c- cc API 05685 API 05686 T e c lm ic a l C o lo rle s s w fr h a it c e t i o on i l , f r o vi m s cos chr Jty om 'V at lO og O ra p S h S y; U /l0 0 see , p a g e s 2I4. a n d 22 fo r d e s c rip tio n H CO API 05687 C o n c e n tra tio n o f benzpyrene in s o lu tio n was 0 .1 5 g ./lO O m l* s o lv e n t. A fte r If weeks th e dosage was in c re a s e d to 50 mg. and th e num ber o f m ice was reduced to l . e nt g lcvae l io is s . ml on 100 phy ./ ra g g o 0 .1 5 om at ;x i'liU X M iJ N T y I H V c l . V i l l r . T l i l i AJ 1 l , l :/V` T JO H O K i'.O L U T IO l'lV , OK ' - .liu /. \ CU iiU L i \ \ \ 'J iv U lO U C . S O L V K Kr oo e +i <a CM e o CM CM +1 XA -A iH - 39 - -d aH +l MD CM O A "O +1 C-- rH ao rH O $ H vO iH ? CA aCoA to CO co fA rH e o ? XA Q CM -d" cm \ Xr--A CO sO CO CO oO CM XA r as ch w y n b io d s o lu t b ta in e n o 1 c 7 rcn P I- .v/ A z f n o b n f o o ti c b ~ !b M Ct ^ * Eh fr2 ^ <: Eh > M o CO O 21 H ft) jj! Eh H CO O O 4h 0 04 P 43 o x> u o D< 4 Cm 4-i 30 Eh C oc E-i =< a c O C oc O O <04 H g fi HgfiH H** i o 4 -P 6 * p ss" O O -H o IP o < CBvO -P aj O -H O dvO -P SJ cer; CO Pi C? &4 ,a\-rHd 14 is- P 0- d 14 CA CA XA XA \\ CA CA \ CA \CO 1 tAI rA- A0-- CM CM CA rH WCM O *H CO i--i o -p a- sD K rH 1 t *H p H O T3 O =CO H < XA B o 0 04 > S4 O H 4h C OC O O fi 6 <h 4 g S4 OOH O cSnO -p at 4 a- at h <4. (A XA \ CA \ c-1 I a- a- CM r^\ rH M O 'rH 00 r--v cm -p orO c: h i 1 4n O X) O 4 O rH Jj vC s o 0 04 P4 0 4-i 4-i C oC O O h g c h f | 04. o (3 o aSO -p at F4hr>--yHat 4h_________ XA \ CA \ r^\ XIA aAcj rH CM e i API 05688 roan tfi ra s ncent lo rle s o o C C E X P E R IM E N T S IN V O L V IN G TABLE 111 THE A P P L IC A T IO N OP S O LU TIO N S OF TO THE S K IN OP C3H MICE .ENZ.PYREHE IN VARIOUS SOLVENTS API 05689 C o n c e n tra tio n o f benzpyrene in s o lu tio n was 0 .1 5 g ./lO O m l. s o lv e n t K X rii'liX M K N l'iJ l l . V n ' ,V J N U 'I 'ilK A 1 T L 1 U A T LutJ U K U O IA J T IU N U U K liE N Z l'Y K E U E I K V A id U U E S O L V E Y !' TO THE S K IN OF C 3II M IC E R E L A T IV E C A R C IN O G E N IC POTENCY, ____ P U C i:o n e o titra tio n o f benzpyrene in s o lu tio n was 0 .1 ^ g ./lO O m l. s o lv e n t. P roduct from a lk y la i io n o f benzene w ith propylene te tra m e r (alum inum c h lo rid e c a ta ly s t) . T en - i+i - ffi s n M O o > i! e-i Q S m P <isob^ pWM O & KJ9! P > < K) K P"^ G <(JPhCSK\h fci M *-- CtJ SO 05 PS on X S cq K s HQfc, 0\ X|HOS <lj O OH SS H H Eh 1K JOHH H < w > a ce o S fc H "3 o H H fc Eh B S few s 1-1 <a s oa w O sO H H p S3 s c H cq S | <; C3 o o p-- ! P CrJ H o on C H Oh P H t o 3 PEh e o a 5h CO a! 1 K <W ty o to w QI+HSi S o Po P Oh H (K S Oh Eh M U Ph cm r rH\0 C'-Pt vO <A rHCM rH HO . HO oo OO . OO O c i +1 r-- +1 o f1 + 1 Is- r + I 4-1 o CO fA <A H . H rH o O o O O oo o A P tR vO rH CM OJ + i +1 +i 4-1 +1 +1 1 CO CA o J" oH A- vO o wrH rH rH C\J r\j vO o rH o rH rH fA <A CM CM CM r*^ n~\ \\ \ \ \\ o XA o o oo o o Oo o OO o o rH rH rH w an O CO 0*1 OP fA XA rH rH rH rH tH o o XA O XA XA XA rH rH CM rH rH rH rH oo O o XA o o xa CM O CM oo rH rH rH <A fA <A o~\ fA (A CO ty S Eh M g N G 0 s CM 0) 0 0 0 G0 C 0 G0 N SN P ea S rH G3 G0 G0 G P G > eq p oP Sh 5 o c --- Nr* sU P N N rH p c' o G P ts UO H r1 O O CZ` , *H PV-OX r-J r-J r^ T5 1 C' 1 Po <! w C0 XCAM o>> o o OM P P-> >a M O Pr P1 ft &H p 00 T) P Q 'V \ rH > CM \ OJ N, \ S4 vC f J -- ** -- - rH rH -- -- &3 2 h pa ppa -------------- -- CO a 1 xr\ - cm c-- o XA _rp XA XA -=r Oo XA NO cm CM CM CM CM CM CM ' API 05690 cm R esidual b y -p ro d u c t o f com m ercial a lk y la tio n o f benzene w ith propylene te tra m e r so lve n t. . l m . O er ./lO ly m g o p $ 0 .1 ene K X PER IM l'J IT S IN V O L V IN G T H E A 1' I ' L l C A T 1011 O F S O L U T IO N S O F rV .P .uZP Y H E U F X U V A R IO U S S O L V E N T S TO THE S K IN OF C3H M IC E T a > M uS 3 M S3 S Jd a Ed ts o O Ph a Cs5! aE-* Qo o; a H a H as >tfa < J Ph ui_ a S3& h h Ecoad m ao a ps Ss \ B i-t a aM Eh* i as s-sa! aoa a> m aaca ao ao m H6.ED S a a S3 o CO - 1*2 - xD -d oo 3 xO "LA 00 CM CM p CM co CA p~ CO CA CO CM CM CM \ \\ CM PA xO -d- PA -d" ca CM CM CA CA LA LA LA \A r-1 rH O o O OOO -d" LA CM CM CM CM l was opy r n p ttliio lrui or s e n in ze n ne be rfe yo p tbieonnz lfa CA CA CA PA C5 a m a Eh Eh S-t m____ M 22 Hjy C<;- &*2,. <: O 43 an i i <h i--i LA'S >> la _ k LA Erl O as a fol <*h co-t OHp Ois *soH C No BJO 43 C C- A- OS S"J H ' s xDI nH <Hi PA TJ HA o^ vO s rH a O dQ P Sh O <*H H P- gO O'<D4n3 gf: g0 c O O ->-1 N CBvO 43 d' P A- cd PA PA PA m LA la U\ LA N\ CM CM \\ H H iH r"\ cn 'S N \ \ xC xO t--i CO1 m rH rH f r^1 , i c- p- ACM Is- xO CM xO CM CA CA PA 43 A- n I *H xO Q OP X> Cfh- Lh co o rl S fi -P * o O O *H d *o +3 ss xO 43 f*- n o4v*ra-t m LA Q xO o9Qp Xi P C-t P4 8o 4-0<3 gS gO 0-4 CSO 43 SM4 ftSf* PA PA LA LA N v: H pH N\ H iH \r\ t CO1 fx- f-- xO xO CM CM API 05691 yo k ioanl roaft t oncen roduct C P API 05692 I IAge o f m ic e a t s t a r t o f e x p e rim e n t, 5 V w e e k s . See page 32 o f T a b le f o r d e s c r ip tio n . rr.v lu c t from a lk y la tio n o f benzene w ith p ro p yle n e te tra m o r (alum inum c h lo rid e c a ta ly s t) . (a p p ro x week 15th heavy 300 n il 100 n il 100 medium 20 a lig h t 20 medium 100 medium 100 - i|4 - NUMBER OP MICE f-C*-U\ro ro ro ro ro iNJ 100 1 100 NOIIVDIUiV *5 naj aovsoa J. STRAIN OP MICE AND NUMBER OP APPLICA TIONS PER WEEK c't m m rn re re pr\ X M cn mB E Xt mB Sr*3N SmC s- uooo o o o jree Xx o a) q and 9 th but o c c u rrin g m ice ). m ice th e r fion C o lo r le s s f r a c t i o n o b ta in e d b y cl-<r.-.r,iatorraphy o f f\F I~ 7 1 , TCC re s id u u m . heavy medium SEVERITY OP EARLY ACANTHOSIS AND HYPERKERATOSIS j EXPERIMENTS TO DETERM INE WHETHER CERTAIN M ATER IALS HAVE 1 R R IT A T IO N A L PROPERTIES S e v e re c ru s tin g com parable to th a t im a te ly double the tim e re q u ire d th e uetw een Wz M N ss aM IRRITATION LIKE ' DODECYLBENZENE o aw r am n m m <D oj m o r o c gj >>>>>>>*> > g O ta swQH KAJh X BOh x McwsAch w o 5M6AOh5h eQXcoCOqc TraqaHl XHqai! n1X--!3KqoOi rOH poH1 Xp+pxXxaECBqcHHi5!t!>jf * p4rraqOHH-HH5$ 4r--n-5 P O XuaB* h'qOoOEO '*siCon?Mi. rqaHH} MI c<l xS Bo rHH PxopsH <qEoL gg -\OrO-VUX'-?N1-. C\J _XUUXX=CON1jN-CBSM<XCOL1- XO qO X X pu rH Pr*4OHH4HS) r*Hrt XrOdHS 'LqE5l <qsoX gg Xor~-'--s. oUvxXOmXBOHbqOCoOiNxwAXHHr<GX?Cxqoo1i--Lt . O\xOoPprct,-H'Vci--rH--oi-I1> XXOrXH1* H M<C1L corl o c XO O Caql) *HxaHl rH -4rC4eHr>t 4EHc-1 XOrP-XC--6Ni?i. OXsXOOrXX&CoqOdiqo--fHli>'X*C-XrrrmTCyoCoPHHHh1-L <cc(CI4rf rH TnCc>1 HdB (L CL CQ < <3 p 00 U\ X UN UN O CO mj CM CM CNJ pH 43 rH O 1 pH rn*\ 0) rO pH rH X CJ CJ CO 55 7s-- r^- R T*1 API 05693 L C X rJ ts rfllK J ilJ i'iJ T O D liT E H M iM Ii W iH '.'TIU cU C E R T A IN .V iA T E U lA L S H A V E 1 U U 1 T A T lO f lA L V K O i'E U T 1 E U - 45 - a: aa 63fe c--Hm AJ rH M H U*N rH IA r--` ir\ rH ;i: vrj.TTTV % H~i a-LVSC-C 5 a VJ oO "I 0 -. O 0 u, " ! :*! M C <* 3 uO a 0? CO a c-. <A nr r^\ A A <lri03|i(-iS5 = ? a o a c - <3 P ^ H c to p h E-t a D <0. -H nT o -T* CO aCA Cl OOu CrHO CO to 44 <--* l--! 0 w N s Si HO c-i to S" rH [>i O cHsOfKaEyhaSpsm > <5<2 y a < a to o a < a. 4pC* *aH 03 a r*4 03 >> q > H rw qrl rH C8 O n ri -t q --4 r* c >as c s or rH u3 wtl - % s s !a rr a or 0r*\ .-wan a-ypota- S>5 >> o q 0 q 0<M>4 >> a q ca M O O i 1 J OO o u\ tr\ or-( rA -3" to, /s, -0 -C 0 > e 00w 03 a p p P CO 'H q q 1-* q t E-i *H *H o a P c H cH 00 --H T* <r. rZJ O p 44 44 44 r a O H-* q p 05 o Ctf O 05 O 05 4C45 a 4-- i-n _a q ^H H q in rH M HH q Ch 0 CO P- M5 <-1 qc pz q : c q i-- H rH *>4 n *rH rt i"' ** ao Co rt p CH 0 44 qh 0p C *r 0 rH qh q S3 q q a ta q * U ast. a q# 0 q q ao S J>> s> ! r-H D-* q -p CL, rH n n rcHa C- rH N q.p N rH G, N 00, M * *\ DO 0 0a a hQ H CO o cO *oH pq 4m H G q O pq Hh riH O q0 -- aq t- pO-H ?O qa OS p rH HO O q 0 p izn 0Q 03 Q-l O H O1 44 P O'M --c -4 n 0w *-1 : 4-" OH H0 -- qq a n O q# r- -P *M q LA 30 4 20 +4 30 p 2 4-3 44 J2 0 Hin OJ> rH LT\ 1--10 O tT\ G O rH OH 0 co a too to -0 CO Is- CO O to h aa aa aaa cas <a CO SS CJ * 1--4 | n ca i o CM -=r ! '_1'\ sC ! O O sC I'**- r-- r- CM Ai CA -J API 05694 EXPERIMENTS INVOLVIN':} A LIM ITED NUMBER OF APPLICATIONS OF CARCINOGENIC MATERIALS m X C2J W oo <O2^QO S3 hH C! -4 ftl H IS 2 2 ^afaoCl, ab W g" 2C3! tKo ^05 S H <X Cd O MO Z a, o O s\ 2Eh H, vO \o - 46 - S3 +1 I sO prA* u\ sO \o M3 \o vC \o vO \ o nO \o cava -d"-d" ror-v CO S3 S3 CM :uu ta E C3 > 3 fc O H &, M E O H a< EtdL H 2Z T3---------<2E2 Ed HSfcO e Som M2 o2 Z o CoM CO o cm oCM O' o CM CM CO <A -CoM OCM fOM O' O H CM Mf\0CM 13 o -C2M a) O ca CJ O Zr^l <CO E2d MJ OH b3 Oa CL C2L E- c- < CoM COM CoM CoM oCM CoM o orH CoM 2H<2EH CO WOHs.<0Z foe 2 < d EDS5 O&,d<Oa2HOMh2aW&202 CM r2A O CM CM CM CM CM rH CJ CA CJ . r2oA >CAJ f2uA MHrCAHJ My 2r^v o * 2oM&h 2wp o2CpO roNH 73 P ccO: i C3oO 2. E 2Eo3 CJ o* 2 P O Cmo ca hC 'O Ms O CKO p Kd* 7> 2 rPH C 2 > 2da S>3 c o 0 -op rH O E iCrM, 3E P70333 3 3oaLpTvo<D-aa hs3coai E t>> owe 3 3 as OroH "'s dO CoMO. C 73 jCa3oOO ? CJe CJ6 2 c 2O H s> O1o S3 W S<D1A W 4-, rpcOSoHo 2M <e EO O jM-dMp <L 2L S Sh -- o N2 XcJ o cO -J COHA o2LCrv0VnOA H2 232 2M2 H rH3 <d c<o g2z w rH r^is - 10rH 1 _(T"+A* H rrH*- N Pf- 10 rH fr- CCf'OM1-'i Ok r"! nOrC>O'- A.-aJ a) i-Et C as rH Pi C3 rrHE0H rM>30 M *H C, 73 as C 2 "H as S3 673 JJ &c rHO M 7>3 rCSOHD M Cu *H oo aas 3CO 3. 3 20 o aErSt c 3 as aS P a a H r c 0 E--, E (9 O P P H 73 PQ P p P 3 3O E PE EH c Pn r- rPH-i 33 PT3"t cQ HCc-) 2 HW 9E*H P33 PV3a >' 2 r*BHH Pu 0 i0--sl 3Q H P Hc_ pu 2 CM TJ P3 rHH P3 2 2`"2i cM 3H P3Oo C *PH 2O 2C.' 20 TJ PM3 23O ot3CroC'J 2 P3M p3c MbC PM 2 PPt3cno 2 P2L* Pcf2 3tMo a co PhCj 2 --pc1 pff 2 rt C9 n f LTN r API 05695 ci 'i M HV ao w C<9O In 505 H 5 1 6 s fcl o H K-a<a! Eh Ka - kl - API EXPERI MENT NUMBER NUMBER PERIOD CARCIN OF OF EXPOSURE OGENIC APPLIC A TO O IL O IL T IO N # HAIR BEFORE REMOVAL WASHING APPLIED PER WEEK CLIPPED BY WASHING AGENT ORIGINAL NUMBER OF MICE F IN A L MAXIMUM EFFECTIVE INCIDENCE NUMBER OF OF TUMORS MICE [T . I./w e e k s ) AVERAGE LATENT PERIOD FOR TUMOR INDUCTION !? $ F .L . in w l^ II Si S i* *11 S i 86 L 01 * 06 (C l) (02) 6 01 vO _=r ** 1 * co H v^O^nMO H O CM O HO 0s' C\J nO oj t- no h nO sO cn o H sO OH OH i--1 iH H rH m rH HCO CO CO CO HCM Cr-Ml CHM #. 0O OOr'l r-cOi~C\ MO CMH r~tOr\\ _dO- no C ontrol - (no w ashing) none no 10 m in . soap s o lu tio n no 1 hour soap s o lu tio n no If hours soap s o lu tio n 0s sO mH r- +ml +'hi o r>- r H C\J H -rdH- ^o hCM yes C ontrol none (no w ashing) o yes 1 hour soap s o lu tio n f\ O Oi--1 y e s 9 h o u rs s oslouat ipo n . API 05696 H C\J See T a b le I f o r d e s c rip tio n * Dosage p e r a p p lic a t io n was 100 mg 1 i - 48 - -EXPERIMENTS ON THE RETARDATION OP TUMOR FORMATION BY WASHING CFW M ICE ----------------------1 API EXPERI MENT NUMBER NUMBER CARCIN OF OGENIC A P P LIC A r O IL t io n s 3 HAIR APPLIEE PER WEEK CLIPPED PERIOD OF EXPOSURE TO O IL BEFORE REMOVAL BY WASHING WASHING AGENT ORIGINAL NUMBER OF MICE F IN A L MAXIMUM AVERAGE EFFECTIVE INCIDENCE LATENT PERIOI NUMBER OF FOR TUMOR OF M IC E TUMORS (T .I./w eeks) i nINDUCTION (S#FJL. w k s j CO O' cn O' O ^ CM +O1' +O +v1O +O1 ' +f*1S CM -hd* _cdn- vCoM rin-i rv-(o H _d- m CM O f'- r-- CM y\ 3- cn m -n v> m in >> 5* > > > I? > o O' co o o _d- in oo -d-r- omco-d-ncM HHCVJHHHHH H r-rlo rn c^\ m CO X) CO CO H H H H r-t pH r--1 H men 305 yes 20 m in . dsetoelruget inot n 20 30b yes 1 hour detergent 20, s o lu tio n 307 yes 1 hour lix lte o ll- 20 detergent2 308 yes 20 m in . w h ite o i l 20 1A I m oi m <n 113 yes C o n tro l none 20 (no w ashing) 309 yes 1 hour detergent s o lu tio n 19 310 yes Ij. h o u rs d e te rg e n t 20 s o lu tio n 311 y e s 4- h o u rs w h ite o i l - 20 detergent2 vD J| 4p.1r\ in CM <n API 05697 1 Table I fo r d e s c rip tio n c o H P SJ rH uO -p C UbO P 43 w ta4o jma CO * oo f y C0S20|s OBQSh3 4J 3 O 5 E-l -H 2 tl rt 9 U yhk 2y Ehh Ky y m2 C0xj 2 rP y vH 0 \--1 y y Pi 2 S4 y y <2 M e /r y kyi Cy y yM H KO >v 0 MESH 2y cO C <* j5 < i-i y ^ <3 ~0 n*y ^~ m(-, sy k:-1 "** yP.-:, Xy 0 0 m 2 y yC> 2 I<SJ: 'a yy ^2 Cl pr-i Ci pH M '^H ` < (-. w 0y 'y ^0r-010 [x3 53 CQ Ch -<g S M PS <rH! CH HPH 1 H H0 myH 0 llrlj y0p <0 O C -'P <$ 1 r-i CO Hr Ky ^2Wcn < yya XS2 j!) 53 0rH MrHD \0 \uP, O0J c CM C0M 20 0 c0 0 30 c3 1"OH 60 *H -GP Pcs c 00 5 0 c --s cl r--Orrp-i1 C i-r--t*Ho-i1 3O d |3 O O '-3- 93 93 00 x r^\ <2 vUO\ nUC\ CMvO vO UN UN vD rr- *1/r-*N. CM OJ \O O O O1--i <rOHw OrH rCM-o0w-T, r^. C0M C^-O CMPSmdT- P G G *0rt t.P GO r3H p 0 T3 '3 0 GO G CO G tc H 0 TG* 3O -gP yC/3 y GO c?d c^- O O G 93 0O ! C f'N O'N cn vUON "N rH -rdH- rH f^N H srGH \ 0 CM OCM CP GO O `H KW G 0 43 .-31 O C/3 2 03 h 0 .c C'- m 0 > CrHO H IP1, H r^, NO vC \O x. 0 , O0 y OJ COM' 0OJ -GP rCOH ts-p --nr-------------- 5^+3 ("> U -0P H3 C0Q r ^ & *3 c0 GO GG 03 0 03 0 f>5 e*\ c~\ 00 G C GO GG vO --^ >- rH C^, 'O J0P C0O -rd Hc 0 CQ *-rpH H 3 SH JH 0 (H -pO -pG G0 fGcO O0 .Cas -P -P r3 a^ ---pi K S G to H R O ,G 0 S aM H00<M 3 M K 3O PX 3 dy >. ? p 0 p0 3C 3 *Ph &c -P -P W H 3300 0H *OH 2 P rH rH P p,ao CQJ* CCmj Ph -H Mp G3, G 0P< H0 rHH . PO 003 L.; c.;=h 0 n a 0 >h 0 0 -d tP tl'i c/!l ^ API 05698 E X P E R IM E N TS ON THE HHTTAH DAT 1 ON OF TUMOR FORMATION 13Y W ASHING CFW M IC E- Dosage p e r a p p lic a tio n was 100 mg a w ^o O^QSH' < z O p S-< fl 2- jJ H e O -n x3 aE-H 2sS QPJ. < J Oh O S HSS. o +c \ i - SO - CM -=r CM cO \ O o r-o co OO H C\J O CM mn >> co co oo oo COH H O O -P C HCM bfl P d P T3 P. P. a) P 'dHn ->H o H t HIhS P. O -rl p a H xm) p sH CO PI >o d H P H t 0 P-. P g< cm E; E<Sph ooo ooo 0sr--HI CM s Ph o CO d co a H p, -P iH3 P a P H *d *d H Vi ao, pa. o i--i CO CO > O .p n H * aM m p PI I p heu hcl, oe to -p CO rrHn API 05699 memorandum September 1, 1953 RECEIPTS, EXPENDITURES, AND AlLCC-TIONS -- API MEDICAL RSSI 1RCH ?UND /^ntributiong Received 1945 1947 1948 1949 1950 1951 195? 1953 TOTAL Broenditures Made Prior to September 1, 1953 Harvard University Toxicological Reviews Cutting Oil Study $ 40,253*73 2,500.00 University of Cincinnati Carcinogenicity Fluorine Study Physielogy of Skin 436,506.91 14,000.00 12.500.00 Philip Drinker Miscellaneous TOTAL EXPENDITURES BALANCE ON DEPOSIT SEPTEMBER 1, 1953 $ 28,900.00 27,800.00 98.300.00 64.340.00 94,840.00 97,460.00 104,2.85.00 79.056.00 $594,981.00 ? 42.753*73 463.006.91 15,300.00 512.17 521,572.31 $ 73,403.19 API 05700 9 ESSO LABORATORIES STANDARD OIL DEVELOPMENT COMPANY P.O. BOX Jl, LINDEN, i.J. research division pH D.\ M D. DIRECTOR E. PH D.. M D. head toxicologist INDUSTRIAL HYGIENE SECTION NATHAN V. HENDRICKS. CH.E. HEAD OF SECTION FRANKLIN W. CHURCH. M S. GEORGE M. WILKENING. M.S. pill ii-s ~j.tr oleum Conpai`7 lesvills- Oklahoma joar Dr. davis: now Chaln5.TR of the SPA Committee, I have prepared the attached report sumrariz 'ing the activities of the Kettering Laboratory as I see then to the present tine, rt would be my intention to present this to the Subcommittee on Carcinogenicity Meeting in Houston as a forr-ol report of cur RPA ConritTee to the Suhcocrittee. Because of the unavailability of tins to cbtair. approval from each of the RPA Ccsrlttee uenbers, I an unable to claim this as a full RPA Committee report. However. X plan to rrrive ih Houston at about two o`clock lionday afternoon, September 26th. and have a reservation at the Shaarock Hotel beginning in the evening. I would be happy to go over this report with my and/or all members of the RPA Committee that evening in order to introduce any corrections or sugges tions that might seem warranted, or, if the Committee so decides, to abandon this report altogether. For any members of the RPA Committee who will not be in Houston Monday evening. I would suggest that they write as their criticisms and suggestions at the Shamrock Hotel and they will be taken into consideration at the discussions which the rest of the Coroittee will hold that evening. I an sending three copies of thia report to Dr. Horten uith the suggestion that if ha h?.3 any consents which hs feels I should have included in this report, or if ho disagrees with our interpretation of the work of the Kettering Laboratory, lie vd 11 be in a position to advise me of the opinion of the Laboratory Konday evsn-ng in Houston. The two extra copies of the report being sent to Dr. Hcrton are ior Dr. Kohoe and Dr. Phair. I realize that this report cannot be a complete summary of all of the work done by the Kettering Laboratory, otherwise it would have to be too long, but I do feel thrt it represents, at least in ay mind, the important observations which have bec-n made thus far by the Kettering Laboratory, and in the summary emphasizes The important points that should be pursued in the work of the Laboratory. It is ~ hope that our RPA Committee, if not wishing to accept this report, will deem it odTissble to prepare a report somewhat similar to this one which can be subTittad to the Subcommittee cn Carcinogenesis at the Houston meeting. If it is agrsoeric with thin Committee and with you, it would be my plan to read this report API 05701 * "s:t.rac3? lubezn-S. css .. _ -rv.,- .vr; -.ernirv: tr.d tier. to :s.rs c piss lade avail- to r.ich ci' the "-embers of the oubccmnittee on Car;:.xog3r..esis following the custon aistir.L* Furthermore, it would be my suggestion that the Subconrittee jt/vksh to consider sending a report siailar to this to the hedioal Advisory 2^ittee with the suggestion that it be forwarded to the Safety Committee of Board of Directors of the API. is i have indicated above, the report which I am sending to you now should be gonsidered only as ry own rough draft of such a report, representing only ry ideas, and subject to the approval of the other members of the RPA Coranittee. t trust that our RPA Committee will be able to come to some agreement on this repoct Monday evening in Houston prior to the meeting of the Subcommittee on Carcinogenesis. Sincerely, R. E. ECKARDT, I!. D. BEEsilk Attachment ee with attachment! Dp* C* H*'Hime Dr* E* K. Linriar DP`M* 9. newqodst Mr* 5* J. Adam* flr. U O.-BWrd; Jr*5 *** H. nsodaraoa Bfof A- SMtay Bsrt<^(3) mu..|6f.- Stroopa^ ?,. it *rs _.v API 05702 z? era committie tiiiMinTs on seitember 29, 1953, :c SUBCOMMITTEE ON CAF.CINCGEMIcIS, API MEEICAl REVISORY COMMI'lTlS cU-^ARY 07 THI API PROJECT ON CAT.CI?OGSNESIS *T KETTERING LABORATORY The original program of carcinogenic work at Kettering Laboratory grew out ,f the results obtained from testing a series of 8 oils. The conclusion of this work, reported on November 5, 19^8, was that all 8 oils affected the skin, jvo of them produced mild irritation, two produced a few verrucous papillomata, one produced papillomas and two questionable cancers, two produced a definite oncer each, and one wee highly carcinogenic. Ab a result of these preliminary studies, the Kettering Laboratory under took a more extensive research program under the auspices of the API Medical Advisory Committee designed'to 1) survey refining processes and materials which sight represent potential hazards from the cancer standpoint, 2) biological testing of Indicated oils and tars, 3) chemical research to determine the raters of the carcinogenic components end to develop analytical techniques for their determination, and, h) an epidemiological program to develop sound infernation regarding the actual incidence of cancer among employees of tha petroleum industry and to determine the efficacy over a period of years of the various safety programs which have been or are being aet up by Individual oil companies. To implement this program, it was suggested that materials from a number of processes b# studied, in accordance with the outline on pages 2 and 3 of the fettering report dated October 27, 1950. API 05703 1 - 2Wnn-accoieratia.? Solvents Ir. order to ha7<j a reference 3ta-,dcrd tc which the results of tests of these oils could be referred, the laboratory undertook & study of the carcino genicity ?f solutions of methyl cholanthrer.s in benzene. For reasons related to veil 'c.owr. dose-res :onss phencsena it wae decided to use the avenge latent period for tumor production as a meaoure of carcinogenicity, the avenge latent period being dsfined as the length of time required to reach 5G$ tumor indices. In the coarse of this work with reference standards, it soon became apparent that C3H mics gave fairly reproducible results, whereas CFW nice gars less reproducible results. It was, therefore, decided to standardise the mouse strain and to use only C3H mice in subsequent tests, even though a considerable number of tests on oils with CTV mice had already been started or completed. In addition, it became apparent that if too strong a solution of methyl chelanthrene is painted on the C3H mice too frequently, the health of the mice, as determined by non-tumor mortality end weight curres, is impaired, end, apparently, their ebility to oroduce tumors is impaired at the same time. In adritlnn, other factors which Influence the hme>lth of the mice, such as infections, infestations with lice, etc., are also reflected in non-tumor mortality sad growth rates, and will seriously affect tutor production, thus unhealthy animals from any cause do not produce tumors as readily as healthy animals, and the srerega latent period is prolonged in the unhealthy animals. Since the average latent period is taken as a measure of carcinogenicity, it is obvious that the apparent carcinogenicity of a material could be profoundly influenced by the health of the animrls, and misleading results thereby obtained. API 05704 - 3a The f.'c're cCEsrv.'ticns -sf necessi"'' jf '.ica cr.ca, iv^co, cr ^areo times per wj-;k c:p.-rd di -.'.u 3'Ter.gth of section being ^eb-, me -o ,.ie .stablishuer.t -f Thr>-:e or -ere 3;:p.ir.- te ftimdircl curves dacer.'ling o;i the frev.uer.cy of painting. Ihe lost recent re ".r- from me Kettering Labcrntorr, however. h : shown that '"hjie riu'=s curves can c-e unitec. ir.`:. a 3ingle curve when th:!j' c.re expressed in ter.ns of the djse of carcinogen applied cor ai ; gi-e*. of skin per week. This has necessitated treasuring the area cl skin covered b7 a solution of methyl nholanthrone in a 6olver.t. A'lthir limits -ns hss been found to be constant for each solvent tested. The solvents thus far tested have been benzene, hydro-peroxide, free aayl benzene, cotter, seed oil, technical white oil and fractions of refinery streams. The effect of these golvente is their effect on the spreading of the solution. Thus if the same amount of the same concentration of methyl cholanthrene in two different solvents were to be painted on the skin ? times a week, the dose per unit area need not be the same. If one solvent caused the solution to spread over twice the area of the other solvent, the does per unit area in the first case would be half that in the second case, and the average latent period for the two solutions would bf ^iffe^rsnt, leading to different values for the two solutions. The Pjjq Onl^ tp. concentration of methyl cholanthrene in benzene, and serw^i. JHjjjLf* a ppj^prence standard. What this work implies is that It "JT ^.carcinogenic work to determine not only the fre quency fldWjjjjtlpotion end the. Mount applied per application, but also the area covered b*fFle amount. API 05705 1 ."olvants Vhft s b=sn said above ap^lieB only to solutions of pure carcinogens in ^..-cccl^ting solventB. In the course of the work at the Kettering Laboratory .. its :bserved that certain advents, in addition to having an effect on the .it. tf 3f to* solution also have a specific effect on the carcinogenicity .? the solution. Thus it was found that diffaran: concantratione of methyl ....ijnthrase in some solvents, even though they spread over the same area, have ,pentid!r the same average latent period. According to the above, this should ,jt be the case since the dose per unit area should be different. In this case e area remains constant, the amount applied per application remains constant, ^ the concentration varies, and hence dose per unit area varies. Theoretically, therefore, the average latent period should vary, but actually this was not found tj be the case. The explanation for thie effect is that the solvent exerts lose specific accelerating action on carcinogenesis. To summarise these two eoacepts, It may be well to give some equations. Thus doss par unit area par weak gjy be defined as followsi 4. mJLS. where > sga of solution applied per application e * soaosntratioa Mvia . r of applications per week Int a# 2 aoa-accalera ting solvents with different spreading, iper. w constant hat a varied, thereby influencing d* In the second a wars constant but c varied and hence d also was variable. API 05706 tout this did not influence average latent period. Hence tvo situations exist, ^ in which the rate of tumor formation is related directly to d, the other in tfjii#1 ^ *8 no** dodecyl benzene and normal hexadecane have been found t0 toe accelerating solvents, although gasoline is not. An interesting observation recently made is that if dodecyl benzene is applied cnee to the skin, and then serial biopsies taken, holes in the epithelium egn he observed microscopically but not grossly. Zt has been postulated by the Kettering Laboratory that these holes provide foci of rapidly regenerating epithelium which may be responsible for the accelereting action obeerved. g^flnery Streaae Ae a result of the above information of a fundamental nature developed over the past several years, tks Kettering Laboratory now believes that carcinogenic refinery stream may be divided into tvo main classes: 1) . Those streams or materials in which PMC is related to d. 2) . Thou streams or materials In which T^g la not related to d In the first group fall' those materials such es eatalytlcally cracked residua in which carcinogen concentration iV'fd'hlgh or accelerators relativsly o low that the earclaapm conoautrmtiou becomes of flrot importance as a measure of tumor activity itts+mIdle meomWI goap, characterised by medium distillates. the accelerator concentration important, la producing tunora bee developed information which indicates that the accelerators are present in those, refinery stream* boiling. f*om 450-650*7.. while carcinogen* ire present in those streams boiling above 650-700*7. Thu in mixtures which API 05707 I a vide toiling range, say from ^50-1000*1., the carcinogenicity of the g^ture nay not he dependent alone on the amount of 700F. + material present, ,,ttt also on the amount and nature of the ^50-650F. fraction. In partial sup >ort of this concept, the laboratory has analyzed a series of efinery streams for their benzpyrene content, in accordance with a method developed Pittsburgh, and have relt ted the determined benzpyrene content to tfc* pkc (ft7Pendix 11 of January - September, 1953, Progress Report). these ^terminations indicate that the % BP/P^g ratio essentially divides into two groups) those that run from 0.30 and higher, and those that run from 0.10 and lower. The first group of materials falls in the first class of refinery aaterii-ls, and the second group into the second class. In three instances (A?X^, 71 and 91) this ratio lay between 0.1 and 0.3, and these three oils nay arrant further study. tables of PMC In the newly distributed tablet of Pjg. values distributed on September 1, 1953. essentially three types of results are givens 1). Those in vhidBfboth the average litent period and the PMC values v s. due W umee^aUa^ var^abXes is the biologieal experiments such am peer heslih. toKimity ef applied materials, etci pw the value is net. These are pretty good experiments from a bielogieal standpoint, but ths laboratory still has some reservations about the accuracy. API 05708 - 7- 3). Thoee la which neither the avert-ge latent period nor the PMC value are enclosed in parentheses. These e: pertinents are believed to be as good a biological experiment as it is possible to obtain. Of the sample* in these tables for which a PHC value is given, 57 fall into the first group, 93 f*H into the second group, and 27 fall into the third group. It is the feeling of this writer (and I hope the remaining members of the RPA Committee) that the 150 samples which fall into type 1 and 2 results can be repeated and brought into a type 3 experiment. Since much of the val idation of the theories of the laborrtory is dependent upon accurate values, it is higily desirable that these PKC values be made as accurate as biological experimentation will permit. Another serious defect of these tables is that in eoas experiments, anlftale have been eliminated from the experiment because the laboratory believed that biologically it was oneound to include them. In several cases the results are based on as few as 6 animals. TIT is to ha desired that no results will be reported unless e minimam numbs/' of12C animals are included in the experiment. *r - .. . The Committee recognise! the validity of throwing out some animals, but feels that where this has't&im^GineAhe experiment should be repeated to include at least 20 ani Other Observe! SJWI WAV*- -- The raiffe&ihg is an effective way of reducing the card r* of ah 'crik*Vb the skin of tho mouse. In general, the sooner the oil lr iBftdr off the better. Limited experiments with white oil wash and barrier creams art difficult to interpret, but the laboratory feels that a white nil rinse prior to washing* or the uae of barrier creams in association with washing, may offer advantages greater than with washing alone. API 05709 d 3 f Th epidemiological data is accumulating in the laboratory, with some 1200 sow having bees re-sorted. No definite trends are as yet discernible but t was not anticipated th:t any such trends would be discernible prior to at Aft 5 7ears experience. It is hoped that the support which the General g^it.tee, Division of Refining has given to the epidemiological study will ^isolate renewed reporting of cases with sufficient medical and occupational stories to permit sose early correlations to be made about 1956. ftgggSL 1. She RPA Committee believes that the Kettering Laboratory is performing uportont work In the field of carcinogenesis of refinery streams and materials. p,fy believe that although a great many practical answers have already been obtained, the work should be pursued because the work of the laboratory promises , to provide many fundamental answers to questions which now exist concerning itrial and occupational cairoinogsnsais. 2. It ie believed* that the wort; with noiwacceIsrating eolvente which relates the carcinogenic respottSoTof a solution of pore carcinogen to the dose applied per unit area per vmeK^W'tba skin la of extreme fundamental importance. The Committee urges be`conducted to establish firmly tha corva 9ftgfe a*-* with accelerating solvents ie also of *sf this should be pursued vigorously In ecmsplhte understanding of the relative role of carcinogens and accele9*tix% ih^refinery etre -a- end materials. . API 05710 * -9- * L, Since much of the correlation of chemical analyses with hiological ^tenCy is dependent upon the accuracy of PM(, value*, the Committee lamento the f&ct thct many of the P^q-values thus far reported are baaed on data which tbe laboratory recognizee as not having been obtained from the beBt biological eXperls^nt possible. The Committee suggests, therefore, that tha laboratory dgrote considerable effort to repeating those experiments in an effort to develop ?MC valueB 8X8 based, on the soundest possible biological experiment. 5. The Committee believes that the work with washing techniques and protective er0aas may nrovids intensely practical guides to refinery hygiene practice and suggests that this work bo pursued as new approaches appear to the laboratory. 6. The Coonittee recognises that the epidemiological studies cannot ba expected to produce significant results until at least 1956, and, therefore*, ofges that renewed efforts at adequate case and occupational history reporting be made during the next several years. It believes that adequate medical staffs can do most to insure the success of this program, and in this connection it acknowledges the stimulus that the General Committee, Division of Befining has given to the attainment of this objective. .7 The laboratory 1ms already determined that a number of refinery streams . ...at-*. ..1*?. *-:. . ,. and materials have a relatively high degree of carcinogenlelty. Although ! i-v fci' iii* '-K l. .. data obtained with agfc*>onaaet be directly transposed to humans, there is . the ec&tm': ` r. . sufficient dgljg lay eggjU^tlonal cancer experience to indicate that the petroleum Jfippta*y Hanoi take lightly the observations of the laboratory on the carcindiKloity of these refinery streams and materials. .8 The Committee reeognises that the laboratory at beat can determine from its biological and chemical work only the relative carcinogenicity of various refinery streams and materials. The determination of the hasard is API 05711 * " 10 - ^pendent upon factors of extent, frequency, duration, and length of exposure. w reconized by the laboratory in Dr. Kehoe'e talk to the members of the j.perBl Committee, Division of Refining the determination of the hazard ie at present a function of the individual petroleum companies. Adequate medical ggA industrial hygiene staffs are essential in these individual companies if the7 876 adequately to appraise the hazard. The Committee supports Dr. Kehoe5s plea that petroleum company managements recognize this truism and make plane for the development of such adequate medical and industrial hygiene staffs. 9. It is apparent that the laboratory hae accumulated sufficient data that publication of results will be deemed desirable in the near future. The Committee recognize* the right of the Laboratory to publish, sad urges that the mechanism tie established so that these publications can be reviewed in a finite period of time. At e suggestion, it It prtfpbsed that this mechanism be so established that review by API --nigemattt^ahd Suggestions and criticisms for the laboratory Area API management^# li^f^within' sixty (60) days. If no criticisms er suggestions are obtstfhWrHfr 'tle^fiiSoratbry at the expiration of this time, the laboratory c*M uiUWWVx forthcoming end proceed with publication. It should bo reed0'' tiiHn'tbr the'purpose of helpful critieisa k o whether1 or not there will be ory elearly states that the That a given occur*. <h? . pstant in - r expoa^:**-, API 05712 y I sU|fliARY OF RECENT PROGRESS IN THE INVESTIGATION OF THE CARCINOGENICITY I OF PETROLEUM INTERMEDIATES AND PRODUCTS 1 September 29, 1953 In the course of the biological program to assay the relative carcinogenic potencies of refinery streams, it has become apparent that some of the materials have a practically constant potency, PMC, over a wide range of conditions of exposure, with these oils, the time required for the induction of tumors is essentially the same whether a small dosage, e.g., 5 mg* per ap plication, or a very large dosage, 100 mg., is employed, so long as the same number of exposures per week is involved. When the frequency of application of such samples is varied, the mean time of appearance of tumors changes in just the manner expected for a solution of the synthetic carcinogen, methylcholanthrene, in a "non-accelerating" solvent such as benzene; hence the constancy of the values of P^g, since these are based upon experiments vlth such solutions as external standards of reference. Other materials from refining operations have exhibited potencies, P^g, which increase as much as five-fold with increasing severity of exposure. While a number of the catalytically cracked oils have fallen into this category, others do not. Hence it has become apparent that a given refinery stream cannot be class ified as variable or constant in its potency, PMg, with variations in conditions of exposure, simply on the basis of the unit from which it was collected. Rather it is necessary to gain an under standing of the relationships between the chemical composition of the oil and its effect on animals under various conditions of exposure. API 05713 ) -2 - Although some concern was expressed, when this problem was first discussed about a year ago, that its complexity was such a3 t0 render it insoluble, the experience of the succeeding months has already shown that such is not the case. There is a mounting record of evidence that the variability of the potency, p^, where it occurs, is related to the extent of the contribution 0f non-carcinogenic but accelerating constituents of the oils. Biological tests on fractions of oils such as the T.C.C. cracked residuum, API-71, and the Technical White Oil, API-80, have demonstrated further that the major part of the accelerators are found in the range, l450-650F., and are mainly saturated hydro carbons, some contribution also being made by alkyl (or alicyclic) derivatives of benzene and naphthalene of a restricted range of molecular lengths. Experiments with pure hydrocarbons furnished by other API Research Projects are being carried out to define the limits of this range more sharply. Chemical research on the carcinogenic constituents of the oils has led to the development of a method of analysis for benzo(a)pyrene, outlined briefly in Appendix I, The analyses for a number of typical refinery streams (Appendix II) show that this compound contributes importantly to the potencies of cracked residua, but to a much lesser extent to those of distillate gas oils. Current efforts are, therefore, being directed towards the characterizations of the carcinogens which are more volatile than benzo(a)pyrene, particularly those which are derivatives of the l;-ringed polycyclic aromatic hydrocarbons. API 05714 I -3- The compounds which were extracted from the catalytically cracked residuum, API-8, by repeated reaction with maleic anhydride jiave been tested on mice (as solutions in benzene). The potencies 90 determined indicate that approximately one-third of the carcino gens of the oil were removed by this method. By chromatographic fractionation, a concentrate (API-8-23d) of these "Diels-Alder" carcinogens was prepared, the potency of which indicates that it contains an equivalent of 18 percent benzpyrene. Further experi ments are in progress to identify the potent constituents of this fraction and to develop a method of analysis for them. The picture which has been developed of the interrela tionships between the five variables, 1. the sum of the effective concentrations of the carcinogens (three important classes), 2. the effective concentration of the accelerators, 3, the severity of exposure (dosage per application and frequency of application), 1;. the relative potency, PM(,, and $, the relative area of the skin covered by a standard weight of the oil in question (as compared to the coverage by the same weight of the standard of reference, methylcholanthrone in benzene), may be expressed algebraically by an equation of the following types PMC * /(0d) [KBPCBP + Vm * K*0*] The function, f(cad), has been found to be of sigmoid form, approaching the value, 1, as ca, the concentration of accelerators, becomes small. Thus, when accelerators do not make a significant contribution, the potency, PMC, is independent of the severity API 05715 I -4- f exposure and depends only upon the dosage of carcinogens |t-ne summation in brackets of the effective concentrations of benzpyrene, "Dlels-Alder" carcinogens, and the third class, as y0t undefined) per unit area of the skin. The relationships between concentration of carcinogen (in g.AOO ml.) and average latent period for induction of tumors by the standards of reference (methylcholanthrene in benzene), have previously been plotted as three parallel curves, one for each frequency of application. It has been found that, if the dosage, . of methylcholanthrene is expressed in the units, weight per unit area per week, the data from all of the experiments are related by the following hyperbolic functions (see Figure I): For the average rate of induction of papillomas. (xpap - 2.8)(d + 0.09) = 8.5 (1) For the average rate of induction of grossly malignant tumors. (xca - 15.5)(d + 0.07) r 6.3 (2) (3) where f = number of applications per week. The relative areas of the skin covered by solutions of methylcholanthrene in other solvents have been measured. The average latent period for induction of tumors by such solutions may be calculated from the above equations, making the proper correction of the dosage factor, d, for the solution in question, so long as the solvent is a non-accelerator, i.e., sec.-amylbenzene API 05716 TI -5oV cottonseed oil. For example, a given weight of a solution of inethylcklantkrene in sec.-araylbenzene covers 1,7 times the area | c0Vered by the same weight of a solution of methylcholanthrene in benzane; hence the dosage of methylcholanthrene per unit area for the former would be only 0.6 that obtained with the latter, assuming that the concentration of methylcholanthrene in weight percent is the same in both cases. Note in Table II (page 30 of Tabular Summary) that, for experiments involving sec.-amylbenzene as the solvent, PMC is approximately equal to 0.6 of the concen tration of methylcholanthrene employed. When solutions of methylcholanthrene or benzpyrene in accelerating solvents, such as cetane or dodecylbenzene, are applied, an appreciably larger area is covered than is the case vith the non-accelerating materials. Hence the relationships between the surface activity of these solutions and their specific Irritating properties are being carefully examined to discover, if possible, something about the mechanism by which they accelerate Ithe rate of induction of tumors by polycyclic carcinogens. Apl 05717 E X P E R IM E N T S IN V O L V IN Q T H E A P P L IC A T IO N O F S O L U T IO N S O F W E T H Y LC H O LA N T H H E N E IN V iW V iK S K API 05718 vAraa covered by 100 mg. x , Average L a te n t P e rio d f o r In d u c tio n o f Tumors (weeks) I APPENDIX I ANALYSIS FOR BENZO(a)PYRENE IN REFINERY STREAMS \ (l) Prepare a chromatography tube by Joining a 300 ml. reservoir I t0 one end of a Corning 39570 HEYXY sealing tube containing a 20 mm., coarse, fritted disc. This should provide a smooth-bored tube, 22 nun. in internal diameter and more than 90 ram. long. ^2) On a 7*5 cm. column of alumina (Alcoa, activated alumina, grade F 20) in the above tube, chromatograph a 100 mg. sample of II oil to be analyzed (added to column as a solution in 10 ml. of ^0:5 benzene-isooctane). Wash first with 100 ml. additional 50:50 benzene-isooctane, yielding fraction No. 1; then wash with 250 ml. of a 70 percent benzene, 30 percent isooctane mixture, yielding fraction No. 2. Fraction No. 1 is discarded. Evaporate the solvent from fraction No. 2 under nitrogen and redlssolve in 10 ml. of C.P. benzene. With a pipette, divide this solution into two equal parts (5*00 ml. each), termed A and B throughout the I following procedures. (3) Iodinate A by this method: (a) To a 2.5 cm. column of alumina in the chromatography tube, add a solution of 0.5 g. of iodine in 5 ml. of benzene. (b) To A, add a solution of 0.5 g. of iodine in 5 ml. of benzene and pour the mixture on the column. (c) Elute the column with 90 ml. of benzene, collecting the total eluate. (d) Remove the free iodine from the eluate by washing with Jtqueous sodium thiosulfate. API 05719 $ APPENDIX I (Continued) jll) Chromatograph B similarly, without the use of iodine. (ij) Equalize the weights of A and B (to W grams) and determine {Ue spectrum (365-1+15 m+i ) of the iodinated A, using B as the blank. 4t wavelengths where B has the higher absorbance, balance the jpectrophotometer on A and plot the absorbance readings as negative fglue s (5) Draw a straight line intersecting the absorbance curve at and 330 mjx . Let the difference in absorbance between the straight line and the curve at 389 nyi be Then, BP s ADBP_ 35 W . Ad 2L w .87 30.5 tile it is obviously not necessary to determine the entire spectrum (difference in absorption between the iodinated A and the hcn-iodinated B) over the range, 365-1+15 to calculate /\Pgp, It is desirable in order to confirm qualitatively the presence of tanzpyrene by the negative peaks at about 370 and 390 mjjL and the Msitive peak of 6-iodobenzo(a)pyrene at 1+0$ mjx API 05720 T APPENDIX II TYPE op nrfTMWRY STREAM API SAMPLE NUMBER PERCENT BENZPYRENE (BP) PMC (100 mg.) catalytically packed Residuum 8 9 12 20 26 43-1 48 50 63 71 82 0.34 0,07 - 0.09 0.1+2 0.12 0.63 0.47 0.32 - 0.36 0,07 - 0.08 0.05 0.10 0.53 0.5 - 0.8 0.06 0.42-0.46 0.16 0.65 0.43 0.22 0.15 0.14-0.19 o.55 0.33 4-BP PMC 0.5 1.33 0.95 0.75 0.98 1.09 1.55 0.50 0.30 0.18 1.61 Thermally Cracked Residuum 6 65 113 0.02 0,06 0.09 - 0.10 0.08 0,12 0.18 0.25 0.50 0.53 Medium Distillate 23 28 33 79 91 102 105 0.02 - 0.03 0.02 0.01 <. 0.01 0.03 - 0,04 0.01 ^.0.01 0.25 0.31 0.51 0.14 0.28 0.19 0.11 0.10 0.06 0.02 < 0.07 0.13 0.05 <0.09 ?avy Cracked Distillate Ui 1.03 0.34 3.03 API 05721 from the Kettering Laboratory, College of Medicine, University of Cincinnati, Cincinnati, Ohio investigative Team: Prank P, Cleveland, M.D. Ralph T. Denham, B.S. Mary Jane Graf, B.S. Francis F. Heyroth, M.D., Ph.D. A. Wesley Horton, Ph.D. Eldon Parkinson, B.S. John J. Phair, M.D., Dr.P.H. Ruth C. Pierle, Ph.D. Helen E. Plagge, B.S. Fred Shaffer, M.S. Charles Stevens, Ph.D. Dorothy A. Templeton, B.S. Russell Tye, M.S. Waldo J. Younker, -M.S. Otto Bufe Marlon Duvall Richard Graeschel Lenore Hull James Pancake Jean Rehm Elizabeth Rush Effie West Report: A. Wesley Horton, fch.D Approved: Robert A. Kehoe, M.D. Director Date: September 29. 1953 Copy from D. V. Stroop for Information of All Members and Associates nf the Medical Advisory Committee October 13, 1953 ESSO LABORATORIES Standard Oil Development Company P. 0. Box 51> Linden, N.J. Medical Research Division Industrial Hygiene Section October 8, 1953 I Mr. D. V. Stroop _ American Petroleum Institute 50 West 50tn Street New York 20, New York Dear Mr. Stroop: I am enclosing a revised copy of the report of the RP (MC-l) Advisory pnmwf **.< nnhnvft.tori on September 29, 1953 to the SuBdCSmTEtee"on Carcinogenicity of the API Medical Advisory Commit* tee. By verbal arrangement vith Dr. Davis it is my suggestion that you have this report duplicated and distributed to all members and associates of the Medical Advisory Committee. Sincerely yours, /s/ R. E. Eckardt B. B. ECKARDT, M. D. REE:11k Enclosure API 05723 REPORT OF RP (MC-1) ADVISORY COMMITTEE SUBMITTED ON SEPTEMBER 29, 1953, TO SUBCOMMITTEE ON CARCINOGENICITY. API MEDICAL ADVISORY COMMITTEE srrMMARY OF THE API PROJECT ON CARCINOGENICITY AT KETTERING LABORATORY The original program of carcinogenic work at Kettering Laboratory gyev out of the results obtained from testing a series of 8 oils. The con clusion of this work, reported on November 5, 19^8, was that all 8 oils affected the skin. Two of them produced mild irritation, two produced a few verrucous papillomata, one produced papillomas and two questionable cancers, two produced a definite cancer each, and one was highly carcinogenic. as a result of these preliminary studies, the Kettering Laboratory undertook a more extensive research program under the auspices of the API Medical Asvisory Committee designed to l) survey refining processes and materials which might represent potential hazards from the cancer standpoint, 2)biological testing of indicated oils and tars, 3) chemical research to determine the nature of the carcinogenic components and to develop analytical techniques for their determination, and 4) an epidemiological program to develop sound information regarding the actual incidence of cancer among employees of the petroleum industry and to determine the efficacy over a period of years of the various Bafety programs which have been or are being set up by individual oil companies. To implement this program, it was suggested that materials from a number of process be studied, in accordance with the outline on pages 2 and 3 of the Kettering report dated October 27, 1950. Son-Accelerating Solvents In order to have a reference standard to which the results of tests of these oils could be referred, the laboratory undertook a study of the carcinogenicity of solutions of methylcholanthrene in benzene. For reasons related to well known dose-response phenomena it was decided to use the average latent period for tumor production as a measure of carcinogenicity, the average latent period being defined as the length of time required to reach 50$ tumor indices. In the course of this work with reference standards, it soon became apparent that C3H mice gave fairly reproducible results, whereas CFW mice gave less reproducible results. It was therefore decided to standardize the mouse strain and to use only C3H mice in subsequent tests, even though a considerable number of tests on oils with CFW mice had already been started or completed. In additon, it became apparent that if too strong a solution of nethylcholanthrene is painted on the C3H mice too frequently, the health of the mice, as determined by non-tumor mortality and weight curves, is impaired, Mid, apparently, their ability to produce tumors is impaired at the same time. In addition, other factors which influence the health of the mice, such as infections, infestations with lice, etc., also reflected in non-tumor mortality Mid growth rates, and will seriously affect tumor production. Thus, unhealthy Miimals frequently do not produce tumors as readily as healthy animals, and the API 05724 -2- average latent period is prolonged in the unhealthy animals. Since the average latent period is taken as a measure of carcinogenicity, it is obvious tbat the apparent carcinogenicity of a meterial could be profoundly influenced by the health of the animals, and misleading results thereby obtained. The above observations led to the necessity of painting mice once, twice or three times per week, depending on the strength of solution being used, and to the establishment of three or more separate standard curves, depending on the frequency of painting. The most recent report from the Kettering Laboratory, however, has shown that these three curves can be united into a single curve when they are expressed in terms of the dose of carcinogen applied per unit area of skin per week. This has necessitated measuring the area of skin covered by a solution of methylcholanthrene in a solvent. Within limits this has been found to be constant for each solvent tested. The solvents thus far tested have been benzene, hydro-peroxide-free amyl benzene, cotton seed oil, technical white oil and fractions of refinery streams. The effect of these solvents is their effect on the spreading of the solution. Thus, if the same amount of the same concentration of methyl cholanthrene in two different solvents were to be painted on the skin 3 times a week, the dose per unit area need not be the same. If one solvent caused the solution to spread over twice the area of the other solvent, the dose per unit area in the .first case would be half that in the second case, and the average latent period for the two solutions would be different, leading to different values for the two solutions. The PwC value refers only to concentration of methylcholanthrene in benzene, and serves only as a reference standard. What this work implies is that it will be necessary in carcinogenic work to determine not only the frequency of application and the amount applied per application, but also the area covered by this amount. Accelerating Solvents What has been said above applies only to solutions of pure carctasgens I in non-accelerating solvents. In the course of the work at the Kettering Laboratory it was observed that certain solvents, in addition to having an effect on the area of spread of the solution, also have a specific effect on the carcinogenicity of the solution. Thus it was found that different concen trations of methylcholanthrene in Borne solvents, even though they spread over the same area, have essentially the same average latent period. According to the above, this should not be the case since the doee per unit area should be different. In this case the area remains constant, the amount applied per application remains constant; but the concentration varies, and hence dose per unit area varies. Theoretically, therefore, the average latent period should vary, but actually this was not found to be the case. The explanation for this effect is that the solvent exerts some specific accelerating action on carcinogenesis. To summarize these two concepts, it may be well to give some equations. Thus, dose per unit area per week may be defined as follows: d = ngm x c x f a where mgm mgm of solution applied per application c - concentration f number of applications per week a area covered API 05725 -3- In the first case of 2 non-accelerating solvents with different spreading, mgm, c, and f were constant, hut a varied, thereby influencing d. rn the second case, mgm, f, and a were constant, but c varied, and hence d also was variable; but this did not influence average latent period- Hence two situations exist, one in which the rate of tumor formation is related directly to d, the other in which it is not. To date, dodecyl benzene, normal hexadecane, methyl naphthalene, cyclohexyl decane, phenyl dodecane, cetane and phenyl decane have been found to be accelerating solvents, although gasoline is not. An interesting observation recently made is that if dodecyl benzene is applied once to the skin, and then serial biopsies taken, holes in the epithelium can be observed microscopically but not grossly. It has been postulated by the Kettering Laboratory that these holes provide foci of rapidly regenerating epithelium which may be responsible for the accelerating action observed. Refinery Streams As a result Qf the above information of a fundamental nature developed over the past several years, the Kettering Laboratory now believes that carcinogenic refinery streams may be divided into two main classes: 1). Those streams or materials in which Pj is related to d. 2). Those streams or materials in which is not related to d. In the first group fall those materials such as catalytically cracked residua in which carcinogen concentration is so high or accelerators relatively so low that the carcinogen concentration becomes of first importance as a measure of tumor activity in mice. In the second group, characterized by medium distillates, the carcinogen concentration is so low and the accelerator concentration relatively so high that the accelerators are more important in the rate of tumor production of the mouse skin than the carcinogens. The laboratory has developed information which indicates that the accelerators are present in those refinery streams boiling from U50-650F., while carcinogens are present in those streams boiling above 650-700F. Thus in mixtures which have a wide boiling range, say from 450-1000F., the carcino genicity of the mixture may not be dependent alone on the amount of 700F.+ material present, but also on the amount and nature of the 450-650P. fraction. In partial support of this concept, the laboratory has analyzed a series of refinery streams for their benzpyrene content, in accordance with a method developed by the laboratory, and have related the determined benz pyrene content to the PMC (appendix II of January*September, 1953 Progress. Report). These determinations indicate that the # BP/Pjc ratio essentially divides into two groups, those that run from 0.30 and higher, and those that run from 0.10 and lower. The first group of materials falls in the first class of refinery materials, and the second group into the second class. API 05726 - k- jables * ^mc In the newly distributed tables of PKC values distributed on September 1, 1953, essentially three types of results are given: 1). Those in which both the average latent period and the values are given in parentheses. It is the feeling of the laboratory that these results are open to serious errors in their'accuracy dud to uncontrollable variables in the biologieal experiments sueh 'as poor health, toxicity of applied materials, etc. 2). Those in which the average latent period is enclosed in parentheses but the Pjg, value is not. These are pretty good experiments from a biological standpoint, but the laboratory still has some reservations about the accuracy. 3). Those in which neither the average latent period nor the PMC 7611168 are enclosed in parentheses. These experiments are believed to be as good a biological experiment as it is possible to obtain. Of the samples in these tables for which a Pj^n value is given, 57 fall into the first group, 93 fall into the second group, and 27 fall into the third group. It is the feeling that the 57 samples which fall into type 1 results should be examined critieally and some of them repeated and brought into a type 2 or 3 experiment. Since much of the validation of the theories of the laboratory is dependent upon accurate Pj-, values, it is highly desirable that these PMC value*.be made as saurate.as.biological experimentation vllL permit. Another serious defect of these tables is that in some experiments, animals have been eliminated from the experiment because the laboratory believed that biologically it was unsound to include them. In several cases the results are based on as few as 6 animals. It is to be desired that no results will be finally reported unless a minimum number of 20 animals are included in the experiment. The Committee recognizes the validity of throwing out some animals, but feels that where this has been done, the experiment should be repeated to Include at least 20 animals. Other Observations The laboratory has determined that washing is an effective way of reducing the carcinogenicity of an oil to the skin of the mouse. In general, the sooner the oil is washed off, the better. Limited experiments with barrier creams are difficult to interpret but the laboratory feels the use of barrier creams in accordance with recognized practice followed by washing may offer advantages greater than washing alone. The epidemiological data is accumulating in the laboratory, with *oae 1200 cases now having been reported. No definite trends are as yet API 05727