Document 6wBvJoxZ0e9OomrODORxYkko

FILE NAME: Oil Industry and American Petroleum Institute (API) DATE: 1953 June-Dee DOC#: API031 DOCUMENT DESCRIPTION: Letters, Memos, Reports, Financial Reports & Other Relevant Documents from June-Dee, 1953  T T M I N c LAEORATORT S t OF KEOic in i-- IDCR * NUt ^ " NATI Oh io UNIVERSITY OF CINCINNATI DEPARTMENT OF FRCVENTIVE M EDICIN E ANO IHOUSTRIAL HEALTH June 26, 1953 CAELE A D O R ISI EETLAS. CINCINNATI TELEPH O NE' CAPITOL U H DMerp. aDrt.mVen. tSotfroToepc,hDniicraelctSoerrvices 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 Very truly yours, ef API 05619 A merican P etroleum Institute 50 WEST 50th STREET NEW YORK SO, N. Y. Mas 30, 1S$3 BUebaircdarosfit yDoirfecCtolnrsela m t! Cincinnati 19, Ohio s h o e w d M d iC i lo authorlty to eentlaue orli 00 lBveetiutloB o f th tee lirity ad nodo o f action c f oerU ia petrolotai jr o d r to fa r th jmt sedioc t a s 30, 195h, accordio to th terse a f th ocrea-- t deted Jaausry 8, 1906, sud on th bealo o f e eo rl-- badisi fo r emersi1ng osyw ois o f 878,283. Open re e d p t o f fowr approvai o f th eontlnaetion o f th projeet m th th lo -- t-- M f r t t Sh ell pa? yen th o r or IM^OQDb Ihe bai-- o f 1 3 6 ^ lee th n e u e n im bel-- la th f r d m imm 3, 1 $ , t * U he e li Jt h 7 % 195*. I t lo Tderstoed th* m j h alow e la th f r d on Ano 30, 195* i l i he g e tte r to th heerleaa Potrei-- Sostituto, end th* m eeeeeo ter th a m a i o f th badisi le to he obllcsted or peni ltheat e r l t t r rth o rla a tlo n . H a / truly 7-- s, AMXXCAI HBH0UUI aSTCUTS Accepted w i Approved1 u n vw sn r (v c z k h ia t z Barough ito Board e f Dlreetors API 05620 ,R< U 0 * T UNIVERSITY OF CINCINNATI DEPARTMENT OP PREVENTIVE M EDICIN E AND INDU STRIAL HEALTH July-9, 1953 CASCE AOORESS K E T L A I, CINCINNATI TELEPHONE. CAh toc U K Mr. D. V. Stroop, Director Department of Technical Services American Petroleum Institute 0 West 50th Street New York City 20 Dear Mr. Stro6p: I herewith acknowledge with thanks receipt of American petroleum Institute's check In the amount of $l|.0,000.00, covering expenditures to be made on their behalf. Very truly yours, ef API 05621 ; 0l L f 0f C|ACIN*TI UNIVERSITY OF CINCINNATI DEPARTMENT OF PREVENTIVE M ED IC IN E AND INDU STRIAL HEALTH August 26, 1953 CARLE ADDRESS: KET LAB, CINCINNATI TELEPH O NE: CAPITOL 1414 Mr. D. V. Stroop, Director Department of Technical Services American Petroleum Institute 50 West 50th Street New York City 20 Dear M r. Stroop: . I hasten to correct an error made in my letter of August 25. The ashmoouuldnthoafveAmreeardica$n40P, e0t0r0o.l0e0u.m Institute's check of July, 1953 Very truly yours, E. R. Portlage,/Secretary to Dr. Kehoe ef API 05622 UNIVERSITY OF CINCINNATI DEPARTMENT O r PREVENTIVE M EDICIN E AND IN D U STRIAL HEALTH August 25, 195 3 CAM LI ADDRESS: RET LAB. CINCINNATI TELEPHONE: CAPITOL M M Mr. D. V. Stroop, D irector Department of Technical Services American Petroleum Institute 50 West 50th Street New Ydrk City 20 4-r Dear M r. Stroop: 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 $JJJi UUU. DOT' 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, -E-. - Ry. F o r-t-l-a-g -e-/S-e-c-r-e-ta-r-y--toDr. Kehoe ef Enc. API 05624 UNIVERSITY OF C IN C IN N A T I KETTERING LABORATORY I A C C O U N T OF----THE AMERICAN PETROLEUM TWaTTTTPne I FOR 2nd Quarter 1953___________ SALARIES (Based on Proportion of Time Actually Spent on Project) Direct Salaries ............... -- ---- ------------------ ------------- .1 U 6 .4 5 *M . Indirect Salaries Histopathological Preparation...................... ____ .25S-X0.Q- Other Services.......................................... ........ 1 1 , 5 1 - 2 5 l 2 1 . Q 5 A . 7 ~ 3 m isc e lla n e o u s e x p e n s e Purchase o f Anim als-------------------------------------------------------------- . 2 7 8 . 4 0 Special Laboratory Supplies....................................................... .3.46 ..20_ Travel Overhead (Proportion of Heat, Gas, Electricity, Steam, Telephone, Genera! Laboratory Supplies, Postage, Annuities, Pensioes, Maintenance, etc____ 7 ,202.84 8 r53Q.QQ T O T A L .............. Balance A valable for Further W ork at End of_____________________________________ 2 7 ,0 4 4 .1 0 Balance Due Kettering Laboratory at End of._____________________________________ R eceip ts______________________________ Expenditures 2nd Quarter 1953 Balance Available for Further W ork at End of.______________________________ 30a494.zS Balance Due Kettering Laboratory at End o f..2 n d -Q.Uag.tflT.________ 5 ,4 5 0 .6 2 ,M * IT U t . I . I ) ) 5 0 ,4 9 4 .7 2 API 05625 EXPLANATORY NOTES CN TABULAR SUMMARY OP BIOLOGICAL EXPERIMENTS September 1, 1Q53 egree of confidence put by the investigators in the apparent potency of the material as indicated by the rate of induction of tumors. In the cases 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 "Final 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 from the growth or mortality data, that the experimental procedure as 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.Pmc A (normal growth and survival) B (some symptoms of abnormal conditions, but reliable estimate of potency possible) X (reliability of results uncertain) (____ ) (____ ) ____ (____ ) API 05627 I For Information Only - Not for Publication API Researcn frojecc i:C-x 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 Methyleholanthrene 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 1 2 6 26 30 31 32 33 35 37 1+3 1+6 1+7 API 05629 - 1- INDEX TO PROCESSES ______ ______________Process___________ ^-catalytic cracking of virgin gas oil Non-catalytic cracking of catalytic gas oil Steam cracking Viscosity breaking Dubbs coiting Batch coking Delayed .`king Maphtka .'forming polyforming ITydroforuiing Thermofor catalytic cracking (T.C.C.) Fluid catalytic cracking (F.CiC.) Houdry ' Cycloversion Solvent extraction . Fractional distillation Straight run distillation Lubricating oil Wax pressing MEK - Benzol solvent dewaxing Filtration dewaxing Desulfurization by catalytic hydrogenation Acid treating Viscosity effects Retorting of oil shale Fuel oil blending Diels-Alder reaction Solvent extraction with concentrated HaS04 Air oxidation Chromatography Appendix Page Number 6 6 9 9 9 10 10 11 II 11 11 . 13 16 16 1 17 18 1 7 , 1 9 , 2 2 , 2 3 , 2 5 19 20 20 20 20 20 20 20 21 22,23 23 21+ 21+ API 05630 V> fvj r\ rv* ' --' >-- ' >-- -- -- --* I--* * -- --* -- I--*-T> CBCH U D O J O j O J C D O . O j C X O l> O j 03CDOOCX)OOCX>COCI>Cj o u j JJ--J a vn.-p~ v_u tv} I--> rv.t--*o O vij OD-g o -Vjj r\}fv)i\jiv>r\ji\>h-o i i i i i i i i a iiiiii iiiiii API amp10 umber* 2 - index 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. C. C. F. C. C. . nitration dewaxing Dilution of No. 8 Chromatography of No. 8 of No. 8 of No. 8 of No. 8 of So. 8 of No. 8 of No. 8 Diels-Alder reaction of No. 8 Chromat ography of No. 8 Dilution of No. 8 of No. 8 Chromatography of No. 8 Solvent extraction with concentrated HgS04 Solvent extraction with concentrated HgS04 Dilution of No. 8 Diels-Alder reaction Chromatography 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 11b 1183 12b 11+ 20 11+ 22 22 22 22 22 22 22 22 22 21+ 22 2? 11+ 21+ 22 23 23 21+ 12 12 16 1215+ 25 25 25 23 11 17 17 9 11 b b 20 lb * Experiments which are complete, including micropathology, are indicated by an asterisk in the body of the tables. API 05631 -3 INDEX TO TABLE I Process t . c. c. Solvent extraction nith concentrated H*SO* Solvent extraction with concentrated Ha304 Non-catalytic cracking of catalytic gas oil Distillation . Dilution of No. 2$ 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 P. C. C. F. C. C. MEK - Benzol solvent dewaxing F. C. C. Non-catalytic cracking of catalytic gas oil P. C. C. Non-catalytic cracking of catalytic gas oil P. 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 Asia treating Houdry Non-catalytic cracking of catalytic gas oil Non-catalytic cracking of virgin gas oil Viscosity breaking Viscosity breaking P. C. C. Polyforming Solvent extraction Appendix Page Number 7,13 23 23 7 17 17 1^ 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 8 8 18 20 11 6 11 8 8,2106 i 8 6 9 9 11 17 API 05632 Y -k - INDEX TO TA3LE I API Sample Mufflber io 31 3C Procesa T. C. C. Chromatography Unromatography won-catalytic cracking of virgin gas oil F C C jeiayed coking solvent extraction .apctha reforming hydroforming + L C Straight run distillation Hmx oeu(wa*erycwe Air Oxidation Air oxidation Dileelar^yeedr e Coking 3team cracking natch coking F C C F. D. C. Delayed coking Catalytic feed to non-catalytic cracking .'Ion-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 uTaecrWiecateing oil X. C. c. ' Delayed coking F C b. Straight run distillation Non-eatalytic cracking of virgin gas oil T. C. C. Fractional distillation Fractional Distillation Fractional distillation Fractional distillation Dilution of No. 111-1+ Dilution of N t. 111-. Fractions! distillation Fractional disciiiation Puel oil olending Appendix Page Number 2112+ 2k b 15 10 1117 11 12 1 1120 221i+ 11+ 15 Qy 10 13 13 10 9,13 o 9 6 10 10 10 19 19 19 19 13 13 10 13 IS 6 12 19 19 19 19 19 19 19 API 05633 API Sampl wnmber lili Ilk* 11111186$ 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 2108 20 2211 ft - 6- TABLE I SKIN PAINTING EXPERIMENTS ON HIGH BOILING SAMPLES A 1I A L I T I C A I D A T A DISTILLATION [2mm. corr, to 76na, PROCESS API SAMPLE NUMBER PRODUCT GRAV ITY VISCOSITY 750 <7Scf 925 >925c E.P. (*) ($) {%) (F.) Non-catalytic 2 cracking of virgin gas oil n 6 n 59 65 n 72 it 9lj- !t 108 Cracked sidestream Cracked residuum Cracked residuum Cracked residuum Cracked residuum Cracked residuum Cracked residuum 21.2 33/100 ssu 5.2 33 26 l+l - 8.5 33/100 32 - ssu 6.6 16 3 .5/100 14-3.8 29.9 26.3 ssu 9.6 37.1/122 SSP 35 37.5 27.5 - 10.If 31.7/122 35 1+8 17 SSF 2.3 1530/130 SSU 32 25.5 1+1.9 Non-catalytic 20 Cracked cracking of residuum catalytic gas oil lf.7 132/122 51 33 16 SSP i Dilution of 20-1 50% oil No.20, No. 20 50$ dodecyl- benzene API 05635 J l - 6- TABLE I SKIN PAINTING EXPERIMENTS ON HIGH BOILING SAMPLES ^-- STRAIN OF MICE AND NUMBER OF API APPLICA 3AJ1PL3 TIONS fUJC3R PER WEEK S CU Q ca <--i O-i Eh < W o 'J H C< O P Oh O pH Q < (as.) 3 10 ORIGINAL NUMBER OF MICE L OG][ CAL DA FINAL EFFEC TIVE NUM3ER OF MICE MAXIMUM INCIDENCE OF TUMORS (T.I./wks.) T A AVERAGE LATENT PERIOD RELATIVE FOR TUMOR CARCINOGENIC INDUCTION POTENCY, (5/oEL. in wks.) PMC ! 2 C3H 1 100 20 7l 0/75 - - 6 * CFW 3 100 59 C3H 3 100 i 65 C3H 3 100 72* C3H 3 100 94 C3H 3 100 108 C3H 3 100 108 C3H 3 5 20 * CFW 3 100 20 C3H 3 100 20-1* CFW 3 100 20 (30) 20 18 (2 1 ) 11 20 9 20 15 6 5 (2 0 ) (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 20 17 100/25 (25.2*3.5) 0.06+*l (53.2*8.7) ( O . o ^ 1) (23.6*3.3) (o.isl;") (15.1-1.8) (31.0*13.6) to.09i;g|) (24.0*5.6) - (36.1*12.5) (1 5 .0*1 .8 ) (1 7 .ij*2 .1 ) 1 (16.1*2.7) 0 ic + a^7 o,15-.05 i Number alive after 63 weeks API 05636 jrr _ 7 _ TABLE I SKIN PAINTING EXPERIMENTS ON HIGH BOILING SAMPLES A N A L Y T I C A L D A T A --- DISTILLATION (2mm. corr. to 76011c, PROCESS API SAMPLE NUMBER PRODUCT GRAV <75 ITY VISCOSITY (*) 750a 925^ >925c E.?, (*) (%) (F. Non-catalytic 23 PCC gas oil cracking of feed catalytic gas oil 26.7 55.5/100 73.5 20.6 SSU .9 636 tl 24 Cracked 5-9 5 1 .9/210 32.3 34. b 33.1 - residuum SSU from No. 23 n 29 TCC gas oil 15.7 70/100 73 27 - 860 feed SSU ft 30 Cracked 8.5 10/210 3t> 38 26 - residuum SSP from Nc. 29 #1 34 Houdry gas 28.0 4 7 .5/100 9 515 oil feed SSU ti 35 Cracked 2.7 2320/100 *3 2G r m residuum SSU from No. 34 ti 38 Total cracking 16.1 2333/100 32 .2 23c3 43.1 charge; com SSU bination unit ?t 39 Cracked 7.8 177/122 3 1.0 19 .8 48.4 residuum SSF ' \ from No. 38 J it 44 PCC decanted 15.2 241/100 31 cl 54.2 33.6 cil feed LSU tt 45 Cracked -1.7 179/122 2o.i| 3 39 residuum SSF from Nc. 44 ! API 05637 u i yg I TABLE I r-- API T SWAuMMPBLEIF 23* 23* SKIN STRAIN OF MICE AND NUMBER OF APPLICA TIONS PER WEEK CFW 3 C3H 2 PAINTING EXPERIMENTS ON HIGH BOILING SAMPLES B I C L 0 0 I C A L DA T A 0 ta rH Oc Si tC0Oa0 J>oi<nH ORIGINAI 0 a< Q <ti NUMBER OF (mg J MICE FINAL EFFEC MAXIMUM TIVE INCIDENCE NUMBER OF OF TUMORS MICE (TI/weeks) AVERAGE LATENT PERIOD FOR TUMOR INDUCTION ( EL. in wksO 100 20 20 100 20 13 100/19 94/28 12,,5l,,3 (18.5x2.9) RELATIVE CARCINOGENIC PCTENCY; PMC o.2 ^ :11 0 .2 5 t\x | 2k* CFW 3 100 19 . 18 100/15 (9.5l.5) > 0.5 360 29* C3H 1 100 20 19 30* C3H 1 100 (2100) (138) 015 34* CFW 3 100 20 19 35 CFW 3 100 20 13 84/30 1(9020//3333) 84/32 100/20 ( 2 3 .3 3 o?) (24.5*4.8) (23o23.3) (16.7*1.5) o .4 o +%1?"l4 0 . 3C;'t'?r; '' -. 0 nq+-.0O2x Ool5Xo02 38 CFW 3 100 19 13 62/58 (4 5 .1 2.6 ) ( .o 5 :;g |) 39 CFW 3 100 29 18 56/24 (21,9X2.7) = hk C3H 1 100 20 16 1+5 C3H 1 100 10 (20) (98) 94/40 (8898//441D (31.1*3.8) (34o2X5.4) (o.25i;gS) (0.2iioOr{) API 05638 TABLE I SKIN PAINTING EXPERIMENTS Oli HIGH BOILING SAMPLES A I A L Y T I c a l 1 TTK" n r DISTILLATION (2mm. corr. to 7( PROCESS API SAMPLE NUMBER PRODUCT Non-catalytic cracking of catalytic gas oil FCC gas oil feed 1 k7 Cracked residuum from No. I4.6 5k Total crack ing charge It 55 Cracked residuimi from No. $k 61 Cracked residuum ft 63 Houdry gas oil feed GRAV I T Y VISCOSITY 750r <750 925 >925c {%) (%) 23.6 56.7/100 87.9 12.1 0 ssu 9.6 11*0/100 76 15.5 8. ssu 33.0 k.2 7 .8/122 93.9 4 1 SSF 18.8/122 55.9 26.1 SSF 2.C 18 6.3 25.6 73.6/100 66.8 18.2 SSU 2 5 .7/100 48.5 37.6 SSU 15 13.9 AF SA* 1Fi i 1 6^ Cracked 19.9 3 1 .8/100 52.5 31.8 15.7 residuum ssu from No. 63 API 05639 TABLE I SKIN PAINTING- EXPERIMENTS ON HIGH 30ILING SAMPLES BIO o q M CAL DATA 1 STRAIN OF MICE S M AND < O FINAL AVERAGE number O M EFFEC MAXIMUM LATENT OF 0<0 JOu ORIGINAL TIVE INCIDENCE PERIOD RELATIVE APPLICA O H SAAPMIPLE* TIONS Q < NUMBER OF NUMBER OF OF TUMORS FOR TUMOR CARCINOGENIC INDUCTION POTENCY, * \) VM3EP per week (mg, ) MICE MICE ;t I,/weeks) (5#F.L. in wk: . PMC Jo i|6# CF'.V 3 100 20 15 IOO/2I4. (17.9*1.5) 0 r + o02 u -.0 1 - k l CFW 3 100 20 li+ 93/17 (1 3 .0+1 ,6 ) C.22+^? 22 S k CFW 3 100 20 16 IOO/36 (25.02.9) 0 .0 7*.0 1 - S S * CFW 3 100 9 8 89/19 (1 3 .32 .2) 0 21+'^^ * -.07 (19) (15) (87/21) - 61 CFW 3 100 20 17 88/26 (15.2+3.8) (0.22+`J|) - 63* C3H 3 100 20 18 89/26 (20.311.9) (0.11+1,02) 63 C3H 3 100 63 C3H 2 100 12 (19) 20 12 (15) 18 100/19 (100/1 9 ) 100/33 (15.712.0) (2 3 .14+2.6 ) 0.19+-01+ "J3 -i7-:$3 - 61;# C3H 3 100 20 13 100/23 (16.811.9) API 05640  I 1I iI -9 - TABLE I SKIN PAINTING EXPERIMENTS ON HIGH 30ILING SAMPLES > A T T T T T T T T 1 c m "FT- DISTILLATION !2mm.corr. to 76c PROCESS API SAMPLE NUMBER PRODUCT GRAV ITY VISCOSITY <75cf (*) 750 925 >925c (*) (%) Non-catalytic 91 FCC gas oil cracking of feed catalytic gas oil ft 92 Cracked residuum from No. 91 tt 93 Cracked residuum 20.1; 57.8/100 80.1 17.9 2.0 ssu 6.1 102.5/100 70.5 27 2.5 ssu 7.9 i+9.14-/100 87 1+.5 8.5 ssu Steam cracking 86 Cracked residuum 3.5 5000/100 51.9 28.U 181 ssu Viscosity breaking ff ! Dubbs coking It 18 Cracked residuum from No. 17 3.1 1123/210 23.7 21+ ssu 52.3 66B Cracked residuum 6.5 18/210 22.1 29 1+8.9 SSF 6?8 Cracked side 1 5 .6 stream from fractionator producing No. 66 81/100 8i|.6 10.7 1+.7 ssu 52a Cracked sidestream a 53 Blowdown oil 21.0 10.5 14.3 .5/100 95.2 ssu 2.5 2.3 203/100 61.3 23.3 15.1+ ssu 1 Distillation stopped when cracking occurred. 8 Peed to unit included catalytically cracked components, API 05641 - ?- TABLE I KIN PAINTINj- ON HIOH 3CILK.0 SAMPLES 830 API SAMPLE fUMBSI STr.rtlX OF MICE AND NUM3ER OF applica tions PER V.EEK bio L C 0 I M Ch =-i < CRIOINAI QO <CL, NUMBER OF (mg.)| MICE FINAL EFFEC TIVE NUM3ER OF MICE C A L DA T A 1 MAXIMUM INCIDENCE OF TUMORS (T. I./weeks AVERA TE LATENT PERIOD FOR TUMOR INDUCTION [SfoEL. in wka, RELATIVE uARCaNC 3-EKICi POTENCY, PM.C 91* C3H 2 ICO 20 19 95/23 (16.4+2.1) i0 j 5 5 - 8801 9 - .92 C3N 2 100 93 C3K 3 100 I 3 (20) 20 11 (14) 18 91/23 (79/23) 94/40 (15-414=1) 1c,33^ 'r:h (29.0 12,8 ) <0.391;?;) 36 C3H 1 100 18* CFW 3 100 10 (20) 10 8 (17) 100/36 (100/4 9 ) 9 56/30 (35.011.2) O.I'Vt.Ol (30.1 +1 1 .3 ) (C.0?,03) 66 C3H 2 100 67 C3H 2 100 20 10 (20) 19 9 (13) 100/37 89/17 (77/17) 2 9 .3+2.0 11.313.2 0,321.01 - 1 - a6 "^'-.13 3 - 3 52 cfv: CFV.r 3 100/15 * 53 100 10 (20) 100 20 8 (15) 20 100/36 (87/36) (18.917.9) 10.211.2 0,1n+ ?7 7 0.53:.cl 1 API 05642 - 10 - TABLE I SKIN PAINTING EXPERIMENTS ON HIGH BOILING SAMPLES A N A L Y T I C A L DA t r ' DISTILLATION (2mm. corr. to 76 API 750 SAMPLE GRAV 750 925 >925 PROCESS NUMEER PRODUCT ITY VISCOSITY ( * ) ( * > (ST) ( I Batch coking 87 Cracked sidestream U+.9 500/100 SSU Delayed coking 28 Cracked sidestream 15.3 361/100 SSU 1+2 .1+ ft 71+ Total furnace 1 2 .5 198/122 22.3 32.5 1+5.2 charge(includ SSP ing recycle) m 33 Cracked 16 .8 1 3 1 .1+/100 60 3 8 .1 1.9 sidestream SSU from No. 7l+ n 81+ Cracked 30o2 55/100 79 19 =9 1 . 1 sidestream SSU n 90 Cracked 30 05 1+0.6/100 90.2 9 =2 .6 sidestream SSU n 96 Cracked 33 =2 1+3 .7/100 777 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 3 1.0 38.1/100 100 sidestream SSU from No = 97 ft 103 Cracked 2 5.2 135/100 1+3=9 35 =1 20.1+ sidestream SSU API 05643 Th l e i SKIN CsFir:;JI-CNE D NUMBER CF SNAAUMPMIP3ELF5 aPpETpRIlOiNWcSEaE-K PAINTIN r 5a ?EF.I..:ENTS ON HIGH SOILING SAMPLES 1 1 i?T M < Id O q m I 0 L 0 1 I C A L DA T A FINAL EFFEC- MAXIMUM AVERAGE LATENT COpO Pini < (&) ORIGINAL NUMBER OF MICE TIVE INCIDENCE PERIOD NUMBER OF FOR TUMOR OF MICE TUMORS 'T.I./weeks) ( INDUCTION 5/^EL. in wks.) RELATIVE CARCINOGENIC POTENCY, PMC 87 C3~ 2 20 20 11 91/30 (23.815.0) (oi0t:ofe) 2d* CFW 3 100 (2100) (1190) 1(80U0//1155) (10.81.6) - k0- . h 28 C3H 1 100 ' (2103.)- (1193) (88154/.3/531) (26.2il+.2) 7k C3H ? 100 (2I k0) (1162) 1(090k/1/ k17l ) (35.8-5*3) - i :l- I o 33* CFW 3 100 20 33* C3H 1 100 20 8U C3H 3 100 20 90 C3H 3 100 20 18 95/15 IO.3+I.3 0 ,^1 -0 .2 2 19 95/30 (19.1+3.U) -5i!:i8 19 95/21+ 16.82.0 *17+*no -.02 17 100/38 (23.3-3-3) 961 C3H 3 100 20 15 93/27 (1 7 .7 i3 .1 ) 97 C3H 3 100 20 15 8?/22 (1 7 .9 -2 .2 ) l o - i 7 * : o k ) C3H 3 98 100 20 11 73/39 (29.57.0) (oii!:Ss> 303 C3H 3 100 20 17 91+/23 (1 5 .9 .8 ) The. pattern of response to the irritational effects of this oil was similar to that observed with dodecylbenzene, API 05644 '( i _ TT TnBLH I SKIN painting EXPERIMENTS on high boiling samples ANA LY T]CAL D ATA DISTILLATION 2mm. corr. to 760a PROCESS API SAMPLE NUMBER PRODUCT GRAV ITY VISCOSITY <750 (JO 75O925>925 E, (50 (30 p Delayed coking 111 V.'ax tailings, $0% benzene 1.2 1 3 1 /21+0 9.6 1+9.8 +0.6 SSF Naphtha reforming t! 19 Cracked residuum 17.0 39.7/100 91 8.95 ssu 0 91 ` 58 ' Cracked 6.3 117/122 35 11+.3 0 8fi residuum from SSF kerosene feed 76 Cracked 30.6 31/130 rj 91 8 8i residuum ssu 1 Polyforming 13 Cracked 10.1 151+/100 63 15 22 3 t residuum ssu 1 f 37 Cracked 11.2 60.2/100 66 15 19 81 residuum SSU 69 Cracked residuum 15.9 1+3/130 75.1 8.3 16.6 SSU 00 I0-- 1 . CO r-- r^\ Hydroforming n 1+9 Cracked residuum 77 Cracked residuum 10.1 a,95 3.5 v9 22.8 A'28/100 100 SSU -3 0o o o 3 Cracked sidestream. 33.2 32.7/100 SSU tl 60 Cracked 21+.9 1+0/100 100 sidestream SSU Distillation stopped when cracking occurred. API 05645 T TABLE I s ^1 " - N ,L2o.pP..) - SKIT PAINTING EXPEPIIvSNTS ON HIGH BOILING SAMPLES S T R A IN CF MICE yjAnNrDarpp OF API APPLICA- TIC"-NUM3EP PFP WEEK cs 30 rr3 M =h E-i < +3 O c w <tO+CL4, O CL, Q < S I 0 Ij 0 G I C A L D A I ORIGINAL NUMBER CF FINAL EFFEC- TIVE NUMBER OF MAXIMUM INCIDENCE OF TUMORS (mg. ) MICE MICE (To I /weeks) A AVERAGE LATENT PERIOD FOR TUMOR INDUCTION (5SSEL. in RELATIVE i CARCINOGENIC POTENCY, ; pMC ! 1+1 C3H 2 100 20 13 85/25 (23.1 2 .0 ) 910 860 60 8901 362 6 - ^91$ 19* CF"' 3 100 10 (30) 0 GFW 3 100 20 (30) 76 CFV.' 3 100 10 (29) 13* CFW 3 100 13 CFW 2 100 37 CFW 3 100 69 CFW 3 100 20 (30) 30 20 (30) 10 (20) 1+9 C3H 1 100 20 77 CFW 3 100 10 (20) 3 C3H 1 100 20 7 (13) 18 (21+) 9 (28) 13 (16) 28 13 (17) 10 (18) 19 10 (19) o1 86/37 (92/37) 89/30 (88/31) 100/30 (96/52) 92/29 (9I+/38) 89/61 lOO/i+6 (91+/1+6) 80/29 (78/30) 81+Al 100/38 (89/1+1 ) 0/62 (26.9+6.1+) (0 ,0 7 1,02) ! (22ol+4.oO) 0,091.02 : 21+.7*3-5 0 .07 .01 (25.63o3) (31 7+54) (254*5.1) (0.07l80ow?J.) I1 o .lt;g j 0,071.02 1 (21+.15.1) (0 o09-03) (35-912-6) 0.18^^ ! (29.3*1+. 8) (0.07!:2) i - - : 60* CFW 3 100 10 8 i (20) (H+) Number alive after 63 weeks. 63/1+8 (57/1+8) (1+34*11.1) (o.oi+l.oi) ; API 05646 TABLE I SKIN PAINTING EXPERIMENTS ON HIGH BOILING SAMPLES -- T T A L Y T I D A L D A T A -- - _ DISTILLATION (2mm. corr. to 760m PROCESS API SAMPLE NUMBER PRODUCT 750 GRAV 750 925> 925* s.i ITY VISCOSITY (%) {%) (*) CP. \ To Co Co 4 ! 109 Cracked sidestream 7 Cra'cked residuum 9 Cracked residuum 2 k -1 U6.1/100 ssu 20.2 31;.5/100 * 1 0 0 SSU 1 8 .1; 37.1/100 SSU 97 2.33 - 780 ti 10 Cracked 11.6 1 1 7 /100 56.7 36.6 6.7 - ; residuum ssu Tt 31 Cracked 16.0 1 4 .8/122 63 35.6 87 residuum SSF n 36 Cracked 21.9 90/100 1+8 892 residuum ssu ti k o Cracked 19 .8 72.6/100 77.2 22.2 .65 3o : residuum SSU ti 71 Cracked 22.8 59/100 77 21.5 1 .5 84; ; residuum SSU * 78 Cracked 16.1 126.8/100 1+5.7 W+.3 10 0 ` residuum SSU Tt 82 Cracked 13 .6 126/130 2 5 .7 51.5 22.8 residuum SSU API 05647 T - 12 TABLE I SKIN PAINTING EXPERIMENTS CM HIGH SOILING SAMPLES -- B I O L 0 G I C A L DA T A STRAIN OF MICE AND number w s0 CL, H E" W c < 0 < M FINAL EFFEC MAXIMUM AVERAGE LATENT OF CO iJ O CL, ORIGINAL TIVE INCIDENCE PERIOD RELATIVE EO,P API APTIPOLNISCA Q P<s NUMOBFER NUOMFBER TUMOOFRS FIONRDUCTTUIMOONR CARPCOTIENNOCGYE,NIC F. sample NUMBER PER WEEK Eg.) MICE MICE (To I./weeks) 5/6EL. in wksj Pm c 109* C3H 3 100 6 (20) 6 (15) 100/21 (93/25) 14.0+4.9 0 pp+20 -.07 7 C3H 1 100 20 9 11/55 - - 780 9* C3H 3 100 10- 9 100A 7 (3602-4.5) 0.06*.01 9* CFW 3 100 10 (2 0 ) 10 (1 6 ) 100/40 (100/4 1 ) (28.4*4.1) 0 07'+-.'0^2 10* C3H 1 100 10 (2 0 ) 9 (17) 100/39 (94/39) (24.6*7.0) (o.4ot;?5) 10 CFW 1 100 10 (20) 8 (10) 88/20 (90/31) (17.3*3.8) 870 31 C3H 1 100 20 17 100/32 (22.0*2.8) 0.40+5)3 0 11 892 36 C3H 1 100 20 16 94/44 (34.9*4.3) 0o22-'.07 360 1+0 C3H 1 100 10 5 100/42 (34-1-7.4) (20) (9) (100A 2 ) 845 71 C3H 1 100 20 20 90/22 (16.6*1.7) 0,55to^ 71 C3H 1 20 71 C3H 1 5 78 C3H 1 100 82 C3H 1 100 20 (33) 19 9 (19) 10 (20) 19 (32) 15 (i h 10 (20) 90/37 (91/52) 0/9 89/19 (86/20) 90/33 (95/34) (27.1*4.8) - (14.3*2.9) (24.1 *6.41 0 .3 1 i:a - -33t l l ' i i The pattern of response to the lrrltatlonal effects of this oil wat similar to that observed with dodecylbenzene,, API 05648 1 - 13 - TABLE I SKIN PAINTING EXPERIMENTS ON HIGH BOILING SAMPLES A N A L Y T I C A L DATA' DISTILLATION (2mm. corr. to 76C PROCESS API SAMPLE NUMBER PRODUCT GRAV ITY VISCOSITY <750 9725500 (50 (50 F. C. C. tl K Cracked sidestream 23 Cracked sidestream 21.6 26.7 55.5/100 78.5 20.6 SSU It 1+6 Cracked 23.6 56.7/100 87.9 12.1 sidestream SSU 1t 88 Cracked 22.5 78/100 69.3 30.2 sidestream SSU H 89 Cracked 27.0 51/100 100 sidestream SSU tl 91 Cracked 20.4 5 7 . 8/ 100 80.1 17.9 sidestream SSU n 101 Cracked 21+.8 40.1 /100 89.7 9.1 sidestream SSU n 102 Cracked 23.5 53.1/100 99 1 sidestream SSU ti 104 Cracked 25.8 60.4/100 7.6 sidestream SSU API 05649 Y 13 TABLE I SKIN PAINTING EXPERIMENTS ON HIGH BOILING SAMPLES B I O L 0 G I C A L DA T A 60mm; E.P, i!p *) STRAIN OF MICE AND NUMBER OF applica API c t ions sflajmMBpEmF PER WEEK W H CtJ 0 O M w ORIGINAL O 04 0 <c 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 k C3H 1 100 20 01 0/69 - - 838 23* CFW 3 100 20 20 100/19 12.5*1.3 - +.11 0 ,2i+-.o5 23* C3H 2 100 20 18 914-/28 (18.5+2.9) 780 1*6# CFW 3 100 20 15 100/21; (17.9*1.5) O'11-:" 900 88 C3H 2 100 20 18 94/26 (17.0+2.4) (o .25!;J) 745 89 C3H 3 100 30 28 89/21 (1 3 .2+2 .0) 830 91* C3H 2 100 20 19 95/23 (1 6 .4+2 .1 ) (O.20i;0) 790 ioia C3H 3 100 20 6 100/12 (i0 .3i.fl) - 700 1 102* C3H 2 20 6 (18 ) 102* C3H 2 100 18 101;* C3H 2 100 20 101;" C3H 2 20 13 (20) 6 (16) 15 19 12 (12) 100/27 (69/43) 93/32 89/26 75/53 (75/53) 22.6+3 .3 (23.8*3.4) 22.4+1.6 (39.Oil0.1) 0 17+.02 u , 1 '-.oi 0 ll+*01 Ooll-.06 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 A N A L Y T I C A L DATA DISTILLATION (2mm. c o m . to 760 PROCESS F e C 'a C < API SAMPLE NUMBER PRODUCT 8 Cracked residuum GRAV <75c? ITY VISCOSITY (*) 11.1 110.3/ 100 IS S3U 9IS2O5 ->925 E. {%) (*) (OPr 22 Dilution of No. 8 8 --4. 50% oil No. 8 , $0% (white oil + West Texas residu 8-16 8-21 50$ oil No. 8, $0% sec-amylbenzene %0% oil No, 0% colorless fraction from No. 8 obtained by chromatography on silica gel________ . API 05651 r - ii+. TA 3LE I .i E.p, SKIN PAINTING EXPERIMENTS ON HIGH 30ILINS SAMPLES -- API samp^ tfljV3ER p m RAIN 0? MICE AND NUMBER OF AP? LICA- TIONS PER V.EEK s 03 O Cd H 04 Eh w o < 1-3 CO Ph O Ph p < (rrtg,,) 3 I 0 L 0 G I C A L D A r a ORIGINAL NUMBER OF MICE FINAL 5FFEC- TIVE NUMBER OF MICE AVERAGE MAXIMUM LATENT INCIDENCE PERIOD RELATIVE OF FOR TUMOR CARCINOGENIC TUMORS INDUCTION POTENCY, !T.Io /weeks) (5#EL. in wksj PMC 8 C3H 3 100 10 9 89/7 (5.1+ii.o) 1 "06-:lo 8 C3H 2 100 20 17 9U/11+ (8.5*2.!+) (.77!^9) 8 CPW 2 100 19 17 100/11+ 7.5-1.8 >0.6 8 C3H 2 20 8 C3H 1 100 8 C3H 1 100 8* C3H 1 100 15 10 (20) 12 (18) 20 13 10 (16) 10 (16) 17 92/20 100/32 (100/3 2 ) 100/25 (100/31) 9 1+/21+ (16.6+1.8) (21.1+5.2) 0.28:;g (0.l+7t"01.) (18.9*3.8) (o.i+i+i;io) (15.9-2.7) 8 , C3H 1 20 8 C3H 1 20 8 C3H 1 50 13 (20) i+o 1+0 12 (19) 32 3i+ 100/50 (100/50) 97/1+2 9iAo (36.l+6.5) 0.2o :;2 (3l.7i2.2) (27.52.2) '23-:o 5 o.29i;!f iduum) 5 8 C3H 1 8 C3H 1 50 - 5 3-4 C3H 3 100 3-16 C3H 1 100 8-21 C3H 1 100 ii+ (20) 33 (1+0) 20 (30) 20 13 (20) 12 (18) 27 (33) 14 (20) 19 11 (15) 100/32 (100/3 2 ) 96/48 (88/1+8) 100/23 (90/23) 100/52 100/27 (67/1+3)__ 2 0 .3 *3 .9 (35-0*3.1) (I8.3 . 9 ) (34.1+3.9) (20.9*3.0) o.20i;i (0 17+0^ ) U#^ - . 0 7 --:?2 API 05652 TABLE I SKIN PAINTING EXPERIMENTS ON HIGH BOILING SAMPLES A NT lT tT : a l D A T A DISTILLATION (2mm. corr. to ' PROCESS F. C. C. API SAMPLE NUMBER PRODUCT 12 Cracked residuum GRAV ITY VISCOSITY 1 8 .1 71/100 SSU 750 <750* 925* >925 (*) {%) {*) ff 26 Cracked 7.3 341;.2/100 6I.3 34.1 4.6 residuum SSU H 42 ' Cracked 14.2 139.8/100 28.6 63.6 7.8 residuum SSU fl ij-3 Cracked 21|.2 Pour Point 24.5 6 7.8 7.7 residuum 110 ft (44) Cracked 15.2 241/100 31.1 54.2 13 .6 residuum SSU W 1*8 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 f ' 85 Cracked 8.1 216/100 42.6 47.8 9.6 residuum SSU API 05653 - 15 - >25` E.Pj TABLE I SKIN PAINTING EXPERIMENTS ON HIGH 30ILING SAMPLES API sampls STRAIN OF MICE AN D number OF applica- TIONS P5R 'VEEK BIOL gB cd u< a m CO A h ORIGINAL O CU o < NUMBER OF (mg.)| MICE 0 G I C A L DA'r a FINAL EFFEC- TIVE NUMBER OF MICE AVERAGE MAXIMUM LATENT INCIDENCE PERIOD RELATIVE OF FOR TUMOR CARCINOGENIC TUMORS INDUCTION POTENCY> (To 1 /weeks' (5#EL. in wks] pMC 1 12 CFW 1 100 18 17 83/3i+ (19.0+5.3) 0oi+6-,21+ i 12 C F V 3 100 15 Ik IOO/U4. (107io9) 26 C3H 1 100 20 19 100/20 (11+,3*1.3) 0 .65^ $ 00 KjJ 01CD0+ (--10 O fV) k2 C3H 1 100 20 18 9k/3k (23.3*2.7) k3 C3H 1 100 20 15 100/37 (33025) *2o-:Sf (W) C3H 1 100 20 16 91+A0 (31.1*3.8) (o.ss!;^) C3H 1 100 20 13 IOO/36 (3 1 .5*2 =2 ) (o .22!;OJ) CO 68 C3H 1 100 20 15 100/17 (U+.8+.9 ) (0,62t*g|) v-n 73 C3H 1 73 C3H 1 C3H 1 100 100 100 10 (2 0 ) 20 20 7 (1 2 ) 19 18 100/22 (83/2 2 ) 100/31 100/25 (18619) (21+.l+*2.5) (1 8 ,1 *1 .6) 0ok5t% l 0.1+7 * .06 _________________ 7 / Hair clipped API 05654 TABLE I SKIN PAINTING EXPERIMENTS ON HIGH BOILING SAMPLES A NALY TI DISTILLATION (2mm. corr. to PROCESS Houdry f! ft API SAMPLE NUMBER PRODUCT 11 Cracked residuum 31+ Cracked re s iduum 50 Cracked residuum 51 Cracked residuum 63 Cracked re siduum GRAV ITY VISCOSITY C 7S 0 (%) 750 925 >925 (*) (*) 1+.3 7 1 /100 SSU 81+ 16 28.0 1+7 .5/100 9 SSU 21.3 61+/100 85 1 5 0 SSU 25.7 67/100 8 3 17 0 SSU 2 5.6 2 5.7/100 U8.5I 37.6 13.9 SSU ti 81 Cracked 22.7 1+0/210 1+0.8 52.6 6.6 residuum SSU Cycloversion 22 Cracked residuum 25.6 814-/100 1 + 9 .5 1+1+.2 6.3 SSU API 05655 T - 10 - TABLE I SKIN PAINTING EXPERIMENTS ON HIGH BOILING SAMPLES -- API sam ple DUMBER STRAIN OF MICE AND NUMBER OF APPLICA TIONS PER 'VEEK 0 r3 w y <03 O M < J CO CL O CL (31g) BIOL ORIGINAL NUMBER OF MICE 0 G I C A L DA T A FINAL EFFEC TIVE NUMBER OF MICE MAXIMUM INCIDENCE OF TUMORS (To Io /weeks) AVERAGE LATENT PERIOD RELATIVE FOR TUMOR CARCINOGENIC INDUCTION POTENCY, (5$SL. In wks,' PMC 880 11* C3H 3 100 20 18 9U-/19 (1 5 .0+2 .2 ) (0.22i;3) 615 3 ll* CFW 3 100 20 19 8U/32 (23o23o3) 820 50 CFW 3 100 20 16 100/21 (15.0*1.7) (.i5t;gf) an 51 CFW 3 100 20 18 83/19 (1 3 .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 (100/1 9 ' 100/33 (20.3*1.9) (0.11io02) (15.7*2.0) (2 3.11*2.6 ) 0.19 ! ; ^ o.i7!f 6 91 C3H 1 100 10 10 100/35 (25.9*11.9) (20) (20) (100/36) 31 C3H 1 - 100 3b 32 91/U1 (30.7*2.2) 81 C3H 1 20 31 28 97/52 (37.7*3.1) 3 22 CFW 3 100 (48> (3D . 7 (10.8*3.1) (20) (Hi) (100/17) ___ API 05656 ji ! I - 17 - TABLE I SKIN PAINTING EXPERIMENTS ON HIGH BOILING SAMPLES ru A l Y T I C A t D A r r DISTILLATION (2mm. corr. to 7 PROCESS API SAMPLE NUMBER PRODUCT GRAV ITY 750 c750 925 >925 VISCOSITY OS) OS) OS) Solvent 16 Phenol extraci 9.7 352/210 62 extraction from Coastal ssu 900-X n ti tt Distillation Dilution of No. 25 17 27 70 75 25 25-1 Duosol extract from Califor nia waxy residuum, in sec.-amylbenzene 7.1 2276/210 SSU 3 28.7 66.3 Phenol extract 1 1 .0 173/100 80 19 0 from San SSU Joaquin naphthenic distillate Nitro-benzene extract from Barbers Hill 60/80 15.7 99 .1+/210 SSU 5 85.3 11+.2 SOa extract from TCC gas oil U+.1+ 38.3/100 100 ssu 700+ bottoms 21+.3 1 1 5 /100 1+9 1+5 6 from F.C.C. ssu heavy cycle gas oil $0% oil No.25 $0% (White oil + West Texas residuum) 2 1.1 3 2 0 .6/ 1 2 2 SSU API 05657 f TABL I SKIN PAINTING EXPERIMENTS ON HIGH BOILING SAMPLES -- API SAMPLE kltIiSB STRAIN OF MICE AND number OF applica tions FER '.VEEK C2 a O a, m e-i -JC o o C M oGOJa, Q a. < frag,) I C L ORIGINAL TOMBER CF MICE 0 0 I cj A L D A T A FINAL EFFEC TIVE NUMBER C 7 MICE AVERAGE MAXIMUM LATENT INCIDENCE PERIOD RELATIVE OF FOR TUMOR CARCINOGENIC TUMORS INDUCTION FCTETOF, (T. I./weeks' (5/tFL. in wksj P m c l6* CFW 3 100 10 6 67/73 (68.3*16.0) (C.03,0 1 ) 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) (0.0;:2+) 27* CFW 3 100 10 (20) 9 (12) 89/17 (83/17) (il,,4+3.0) '<L 70 C3H 3 100 (20) 5 (11*) 100/42 (93/1*4) (31.2+11,5) r, Oo+.04 ' *-.05 75 CFW 3 100 20 13 77/51 (38.67.2) (0.0?t,01) 25* C3-: 3 '100 30 in 95/19 (13.4*1.4) -+.03 J 05 25-1* C3H 3 100 30 26 92/40 (27.03.7) 0 '0+"01 u,J-~,03 API 05658 TABLE I SKIN PAINTING EXPERIMENTS ON HIGH BOILING SAMPLES A N A L Y T I CAL D A T A^ PROCESS API SAMPLE NUMBER SPECTRUM TYPE OR PRODUCT DISTILLATION (2mm. corr< to ye 750 GRAV <750 925 >925 ITY VISCOSITY (%) (?) (?) ( Straight run distillation n 1 Distillate # 11896 31.9 56 Waxy Midcon 29.9 tinent dis tillate, Type .B ( .5) 37/100 SSU 87/100 52.u U1.6 (2.0) SSU ft 79 San Joaquin 16.6 63.9/210 rJ 13 86.2 naphthenic SSU distillate, Type C (2.2) *T 105 West Texas 28.9 5U.3/100 72.9 2 3.U 3.3 paraffin SSU distillate, Type B (. 5) tt H U Type B (.63) 2U.6 235/100 SSU 1 115 Type C (.58) 2 3 .2 23U/100 SSU tt 5 Residuum from 12.8 572/122 26 26 U8 Wilmington SSP crude # 11885 See page 8 of Section A of report dated April 5? for discussi of the classification of Straight Run Distillates by Spectrum TyP API 05659 T LASLE I 76P ^ J ( #EP0.p Sr I>' PAINT 7 ` ' - -> I'- 2 ' :-:gh .jcil i:;g s.'.i.FLxs f-- a:.1 D NUMBER OF API a p p l i c a sample tio ns JIJM3ER PEP '-EZK r I 0 l c : : r*- - -1 J ?A 1 ^ c5 sa &0 Ph ORIGINAL 0 9, NUMBER c < OF (ng.) MICE FINAL Er FEC- TIVE NUMBER OF MICE : l da MAXIMUM INCIDENCE OF TUMORS (To I,,/weeks T A AVERAGE LATENT PERIOD RELATIVE FOR TUMOR CARCINOGENIC INDUCTION POTENCY, (5/EL. in wks,) PMC 1 C3H 1 100 20 1 0/72 - - 830 855 56 CFVV 6 56* cfv; 3 56z C3H 3 56 C3H 3 79* C3H 2 100 20 100 10 (20) 100 ` 20 * 20 20 100 10 (20) 15 8 (17) 20 20 7 (14) 87/22 100/20 (88/22) 5/ll 0/1$ <?$?, (15.3*2.9) (15.4*3.2) - - 0 - + 0 11 u " lS-.05 - - - (39.2+15. It) ( o . i k l - X l ) 105s C3H 3 100 14 (20) 13 (17) 85/33 (83/33) (2 3 .2+4 .4 ) 111; C3H 3 100 20 14 0/23 - - m C3H 3 20 20 14 7/43 - - 115 C3H 3 20 20 20 5/26 - 5 :3H 1 100 20 o1 0/56 Number of animals alive after 63 weeks s Hair clipped The pattern of response to the irritational effects of this oil was similar to that oLserved with dodecylbenzene API 05660 - 19 - TABLE I SKIN PAINTING EXPERIMENTS ON HIGH BOILING SAMPLES A N A L Y T I C A L D A r a ** DISTILLATION (2mm, corr . to PROCESS SPECTRUM API TYPE1 GRAV 750 <750` 925 >925 SAMPLE OR SA NUMBER PRODUCT ITY VISCOSITY ($) ($) ($) N Fractional 110-1 Type 3 (.001*) 61.0 .95/70F.a 100 distillation ft 111-1* Type B (1.62) 23.0 17/11*0F.a n 111-1*6 75$ No. 111-1* 22.7 2.5%. Benzene n 111-5* Crude residuum 15.1* 112/100f ! 11 (5$ Benzene) Dilution of No, 111-1* 111-6 50$ No. 110-1 50% No. 111-1* 11 111-7 50$ No. 111-1* 1 50$ No. 112-1 Fractional 112-1 Type B (.016) 65.9 ,896/70oF.8 100 1 distillation 112-3 Type B (.01*5) 39-7 3 .i*/ll*0Fs Lubricating 99 oil rt . 100 i* 100-1 Type C (.5) 20.0 Type B (.3) 29.5 0.2$ Antiox idant3 in NalO! l(1*5725//120100 n) 38 V ssu f307/100 <53/210 V 0 ssu 4 100-2 0.5$ Antiox idant, 2,6-di- tert. -butyl-1*- See page 8 of Section A of report dated April 5? 1952 for discussi<*| 2 of the classification of Straight Rum Distillates by spectrum Kinematic viscosity (centistokes) 3 Sams type additive as that contained in API-99o API 05661 T - 19 TABLE I SKIN PAINTING EXPERIMENTS ON HIGH BOILING SAMPLES API sample tfUMBER STRAIN OF MICE AND NUMBER OF APPLICA TIONS PER WEEK os < uj CO &H O Oh Q < (mg.) BIO L 0 G I C A L DA ORIGINAL NUM3ER OF MICE FINAL EFFEC MAXIMUM TIVE INCIDENCE NUM3ER OF OF TUMORS MICE [T.I./weeks) T A AVERAGE LATENT PERIOD RELATIVE FOR TUMOR CARCINOGENIC INDUCTION POTENCY, :5EL. in wksj PMC C3H 3 20 20 191 0/55 tm IT iii-U*C3H 3 20 19 17 8 8 /2 2 (1 7 .6lo6) 0 i 7+ 02 U#-L '-.03 111 C3H 3 5 20 20 0/3 - - m-a C3H 3 5 20 20 o /5 - tm I 111-6*C3H 3 20 111-7tC3H 3 20 112-1 C3H 3 20 . 112-33 C3H 3 20 99 C3H 3 100 100 C3H 3 100 100-1 C3H 3 100 C3K 3 100 13 (19) (2 70 ) 20 20 (2I k0 ) 20 20 20 13 (19) 7 (17) 171 181 11 (16) 191 20 19 77/31 (58/31) 100/27 (76M) 0/58 0/58 8 (26.0iij.o5) 22.i;3.5 = - 91/36 (9k/k7) 0/80 (3loOi;.5) tm 0/3 - 0/1+2 0 ol0io03 0 8ll-+#.0<^2 - mt tm .1 1 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 DCILING SAMPLES A i: A L Y T I C A I D A I A ' DI STILLATION (2mm. corr 0 76 PROCESS Wax pressing n MEK - Benzol solvent dewaxing API SAMPLE NUMBER PRODUCT GRAV ITY 32 Dewaxed paraf fin distillate fraction from Lima crude 57 Dewaxed oil from No. $6 28.7 4-3-1 Dewaxed oil from F.C.C. decanted oil No. 1+3 12.9 VISCOSITY <.750 (%) 750 925 >925 E (%) (%) ( 9 1 .3/100 80 195 ssu 195/100 29.5 57 ssu = 5 13=5 Filtration dewaxing of No. 8 Desulfuriza tion of 8-3 by catalytic hydrogenation 8-3 Filtrate oil, 11.3 85^ of orig inal (o9k% S) 65 21.1+ 13 =1+ 21 Hydrogenated 13.1 50.1/100 7i+ heavy gas oil ssu ( . 1 5 % s) 20 6 Acid treating 62 Hydrolyzed 11+.2 7 2 7 7 /1 0 0 58 acid sludge ssu Effect of 111+ viscosity on ' tumor induction Straight run distillate, see page 18 of Appendix 2i+. 0 235/10 0 ssu Retorting of oil shale 134-1 95% API-111+, 5>% alkylpolystyrene 116 Crude shale oil 25=1+ 1 7 5 6 /1 0 0 ssu 19 =1+ 291+=9/l00 ssu API 05663 A :c n 125 E.p, >) TABLE I SKIN PAINTINN EXPERIMENTS CN HIGH BOILING SAMPLES ,, T OF VICE . AND NUMBER OF applicaAPI tions flTMPEH PEP '7EEK j '; P-, tc < ,e E IC j C G I a]^ Pu O H -1 ORIGINAL 5 < <i 0 NUMBER CO M 0 -3 OF Q pL, MICE FINAL EFFEC- TIVE NUMBER OF MICE C A L DA T A MAXIMUM INCIDENCE OF TUMORS ( T d o / w k s .) AVERAGE LATENT PERIOD RELATIVE FOR TUMOR CARCINOGENIC INDUCTION POTENCY, !5/&FL. in wks,! Pmp 32 CFV(73-6 )1 10 0 10 (2 0 ) 6 (15) 1 0 0 /1+0 (87/1+1+) (31.5*1+.3) ,5 57 CFW 6 d9a| l .5 C3H 1 C3H 2 100 20 17 100 10 ` 10 (2 0 ) (19) 20 20 19 # 1--11 1--a1 r^v -=* - 1 3-3* C3H 3 100 20 20 3 - 1 21 C3H 3 100 20 20 100/25 (16,9*2,0) - 1 0 0 /21+ (95/25) o/5 (1 9 ,5 +2 ,2 ) 0 .1+3 * ,0 8 a 90/17 (11.2+1.7) (.36:;07) 100/13 ( 9oi+*lol) (0.1+3*??) 0^ X 350 62 C3H 3 Ilk C3E 3 100 10 (2 0 ) 20 20 9 (16) 11+ 33/79 (25/79) 7/1+3 (98,3*26.5) - Ilk C3H 3 100 20 11+ 0/28 - - 114-1 C3H 3 20 20 15 33/1+6 - - 114-1 C3K 3 100 20 13 0/28 - L116 C3H 3 100 30 20 25/192 - - Three applications per week for 11+ weeks, *eek- t h e r e a f t e r , Pant ing d i s c o n t i n u e d a f t e r 1i 3' wv,,-eee.kirss,. six applications per API 05664 - 21 TABLE I SKIN PAINTIN''- EXPERIMENTS ON HIGH BOILING SAMPLES ANAL YTICAL DATA DISTILLATION (2mm0corr. to 76g. PROCESS Fuel oil blending API SAMPLE NUMBER PRODUCT 113 Industrial fuel oil GRAV ITY 8.3 750 750d 925 >9251] VISCOSITY (%) (*>) (%) 30/122 U3 37 SSF 116 Jatalytically cracked resid uum component of API-119 119 10j API-118, 23.3 9 0 % light cat alytic cycle cil (b,;+50 600) 35. 3. d/uo ssu c ,,c Id,; API 05665 TABLE I SKIN PAINTING EXPERIMENTS ON HIGH BOILING SAMPLES API SAtfPLi \TJM3EP STRAIN O_h 3O CP MICE W M Oh H AND NUMBER S 5 OP w S APPLICA O cu TIONS o < PER WEEK (mg.) B IOLOGICAL DATA ORIGINAL NUMBER OF MICE PINAL EFFEC TIVE NUMBER OF MICE AVERAGE MAXIMUM LATENT INCIDENCE PERIOD OF FOR TUMOR TUMORS INDUCTION T.I./weeks) (5#EL. In 1131 C3H 3 20 2 113 C3H 3 20 3 113 C3F 3 20 3 113 C3H 2 20 3 113 C3H 1 20 3 113 C3K 3 100 3 113 C3H 3 5 3 113 C3K 5 1 7 (27) 11* (27) 20 (27) 7 (27) 27 27 ( 1*0 ) 26 (39) 20 7 (22) 10 ( 16) 20 (25) 7 (22) 20 27 (36) 26 (38) 18 100/25 (77/27) 100/28 (83/28) 100/25 (100/25) 100/ 21* (91/1*0) 95/1*5 100/23 (100/ 25) 100/26 (92/31*) 78/51* (1 8 .83 o8) (2 2 .1 *2 .5 ) lm - M ((1188.. 11+11..65)) 22,,5-6 (26.6+3.5) (35.C2.6) (16.3*1.3) (17.5*1.3) (16.9+1.5) (13.9*2.1) 1*6.1 RELATIVE CARCINOGENIC POTENCY, pMC 0 . 16+ . 01* n -,k +*6| 0s1D-.03 0. 16*.02 o.i9.+:gl n ifl+ Ool8-.01 0.17*.02 Ayo.Ol* 3 113 C3H 3 20 20 4 113 C3H 3 20 20 16 100/27 (19.2*2.1) n0 1)77-+,,0011* 20 100/26 (1 8 .3 *2 ,1 ) QL "i-^+".0032 118 C3H . 100 1 2/month) 21* (30) ( 2211+) ICO/50 (100/50) (36-.9l*.3) a i + .02 Ool+b" .17 -19 ( C2 .3/ HT O r.th) 100 119 C3H 100 (2/we ek) 29 22 (27) 25 19 (23) l*o/5i* 95/27 (96/30) 21.6*1.5 O.I8 + .02 ,01 , Approximate age of mice at start s Approximate age of mice at start Approximate age of mice at start ' aximate age of mice at start of experiment, of experiment, of experiment. of experiment. 27 weeks, 23 weeks. 20 weeks, 11+ weeks. API 05666 - 22 TABLE I SKIN PAINTING EXPERIMENTS ON HIGH BOILING SAMPLES PROCESS AFI SAMPLE NUMBER PRODUCT Fractional distillation ft 1 tt -61 0 - 9 .3% fraction from API-8 b-71 8-8 1 8-9 1 9 .3 - I9 .I4.J0 fraction from API-3 IQ.I4. - 3O.35& fraction from API-8 30.3 - 39.8/0 fraction from API-8 (i rt-1 0 1 39.8 - l4.9 6# fraction from API-9 ESTII/ATa CORRECT^ o CILLjg RrtuGE (Id F. a ~'0<> 3Z k25 - 5 8- 560 - b 63O - 95 - 7 710 - 7 M 8-1 1 1 I4.9 . 6 - 59.2$ fraction from API-8 765 - 3 8-1 2 1 39.2 - 69.156 fraction from API-8 800 - 8 Diels-Alder reaction Fractional distillation ft 8.1 2 a1 non-adduct from reaction (9 5.1$ of 8-12) 8-1 3 1 69,1 - 73.5/6 fraction from API-8 J30 - 8-llj.* 78 .5 - 80.8$ fraction from API-8 650 - 3 n 8-1 5 1 Residue; 30.8 - 100$ y 075 ti 3-18 Proportionate reblend of distillation fractions, 3-6 through 8-15 O- solution in benzene. 3 3 .3f; solution in benzene, API 05667 1 - 22 = O 0 ED EO'D Cl 95 710 :r 830 -50 j- h - < J? *17 Cs TABLE I SKIN PAINTING EXPERIMENTS ON HIGH 30ILING SAMPLES STRAIN OF MICE AND NUMBER CF sUaAmMPBpIElHe APPETPRILCICM"ASC-ZK 0PCaLd5. EHO5*H! to<Oo tOP+LJ<. (mg.) B I O L 0 G I C A L DA ORIGINAL NUMBER OF MICE FINAL EFFEC- TIVE NUMBER OF MICE MAXIMUM INCIDENCE OF TUMORS [To I ./weeks) T A AVERAGE LATENT PERIOD RELATIVE FOR TUMOR CARCINOGENIC INDUCTION (5$F,,Loin wksj POTPEMNCCY, 8-6 C3H 2 100 20 H+1 0/66 - 8-7 C3H 2 100 20 161 0/71+ 8-8 C3H 2 100 20 lb 1 0/71+ 8-9* C3H 2 100 20 18 56/62 (58.0+13.1+) - 0 8-10* C3H 2 100 20 16 91+/1+1+ 31.2*3.4 0 0 3-11* C3H 2 100 20 20 90/33 (21+.62.5) 0 00 0 0 1 0 f+'V-l1--1 0 0 0 0 +xnI1--1 8-12* C3H 2 100 8-12a* C3H 2 100 3-13* C3H 2 100 3-11+* C3H 2 100 3-15* C3H 2 100 3-18 C3H 1 100 20 16 20 20 20 10 (15) 11+ 16 11+ 12 13 10 (11+) 93/28 100/26 79/25 25/21 92/35 90/18 (87/39) (19.5^3.1) (17.6*2.1) (22.1+2.5) (26.1*4.7) (28.0+2 .6 ) 13.32.5 0 2 l"i"<1^ Oo3-.06 0 2c+0^3 Uo5=.05 (0.20^q |) - 0 72+"2^ U o 7 " .16 / o o L Nuaber of animals alive after 1+5 weeks API 05668 - 23 TABLE I SKIN PAINTING EXPERIMENTS ON HIGH BOILING SAMPLES PROCESS Diels-Alder reaction #1 If ft tt ft fl tl API SAMPLE NUMBER PRODUCT 8-23 a Crystalline material from 1st cnromatographic frac tion of Class A 1* adduct regenerate, ,3k% in benze o-23b non-crystalline residues from 1st and 2nd chronat graphic fraction, 1 .$% in benzene 3-23c Crystalline material from 2nd chromatographic fra tion of Class A 1 adduct regenerate, in benze 8-23d 3rd chromatographic fraction of Class A1 adduct regenerate, 0.8# in benzene 8-23e 4.th chromatographic fraction of Class A1 adduct regenerate, .$1% in benzene 8-23f 5th chromatographic fraction of Class A 1 adduct regenerate, ,,57J6 in benzene 3-23g Class B adduct regenerate, 1.5/6 in benzene 1 , 8-23h Total Class A adduct regenerate, 2 ,2 % in benzene Solvent extra tion with cone. HaS0* 1 8-19 8-20 Extract from API 8-10 and 8-11, $0% benzene Raffinate from API 8-10 and 8-11, $0% benzene tf 23-1 Extract from API-23, 20% benzene tf 23-2 Raffinate from API-23, 20j 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.), 70# benzene 1 Class A aiduct was tnat normally soluble in aqueous NaOHj Class adduct was not soluule in either $% aqueous NaOH or benzene s^er hydrolysis. API 05669 - 23 - TABLE I SKIN PAINTING EXPERIMENTS ON HIGH BOILING SAMPLES API AMPL3 timber STRAIN OF MICE AND number OF a pplica tion s PER WEEK b i o :1 0 G I a 05 o ril M s* -i <! W O O M < J ORIGINAL w a, C A* NUMBER Q < OF (mg.) MICE FINAL EFFEC TIVE NUMBER OF MICE CAL DA T A MAXIMUM INCIDENCE CF TUMORS (T -Io/w eeks] AVERAGE LATENT PERIOD FOR TUMOR INDUCTION ( S e/oEL. I n w k s RELATIVE CARCINOGENIC POTENCY, P mc 5-2 3 a C3 H 3 15 20 17 o/Sk* - 3 - 2 3 8 C3H 3 3 -2 3 c C3 H 3 6 2 3 d C3 H 3 8-23e C3 H 3 8 -2 3 f C3 H 3 is 20 11 12 10 _ 6 15 Ik (20) 15 9 15 20 13 (16) 8 16 100/51 o/ 5k a 100/36 (100/38) 13A 6 4 o/ Sk4 AJ 3 8 . I - 2 9 . 1^ 2 . 7 - a) O oOS - 0o08o01 > = - S-2 3 g C3 H 3 15 lij- 13 92/50 k 3 S~k2: O . O l + l . O l 3- 23h C3 H 3 15 13 12 100/1+3 ' 3 5 o2 3 , U - o7 - : 2 3 - 1 9 C 3 H 2 100 9 9 22/32x - - 8-20 C3 H 2 100 6 6 ( 2 2 n8 5 c 0 ) 0 .1s: $ (9 ) (9 ) M) \J s 23- 1 * C3 H 2 100 20 19 90/26 (18 .2 *3 .1) 0 o 2 5 -,,,0 9 3 - 2 * C3 H 2 100 20 19 100/1+1+ 1 ( 2 5 7 3 l ) o a 6 -!o i ` 2 -7 C3 H 2 20 20 18 33A 6* -------- t Painting discontinued after 2o weeksi 8 30 weeks; 32 weeks; 5 jjj weeks 0 - - y? weeks; API 05670 - 2k - J TABLE I SKIN PAINTING EXPERIMENTS ON HIGH BOILING SAMPLES PROCESS API SAMPLE NUMBER PRODUCT Air oxidation (cobalt naphthenate catalyst) 83 83-1 Two hour oxidation product Four hour oxidation product Chr oraatography 3-5 Aromatics of No. 8 from Silica Gel a .1 ,, ft 8-1 3 a First aromatic fraction, 2% , of API 3-13 tf d-1 3 b Second aromatic fraction, 2.k%1 of API 3-13 If 8 -1 3 c Third aromatic fraction, 8.7%*, of API 8-13 8-1 3 d Fourth aromatic fraction, Z.6%1, of API 8-13 tl 8-l3e Fifth aromatic fraction, 5.6%*, of API 8-13 It 8-13f Sixth aromatic fraction, 2% 1, of API 8-13 tt 8-13g Seventh aromatic fraction, 16.9% , of API 8-13 It 8-1 3 h Eighth aromatic fraction, 2.k% , of API 8-13 1? 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 3k of Section A--1+ of report dated April 1 9 5 2 *for description tt 71-2 Proportionate reDiend of all fractions from cnromatograpny of .PI-71 It 8-21+ Non-aromatics of No- 3 from alumina Tested at this percentage by weight in benzene, 2 Proceeding colorless non-aromatic fraction, k8% of API-0-3, proved to bo a non-accelerating solvent when diluted 50% with benzene, See API-266 on page 37. API 05671 w^ ... TABLE I SKIN PAINTING EXPERIMENTS ON HIGH BOILING SAMPLES TRAIN OF MICE AND number OF applica- API TIO:IS 5A.PI-E iOMSER PSR WEEK BI O L 0 G I C A L DA a, 0 , a m E<h eq 0 C5 H < ORIGINAL op a. NUMBER a < OF (mg. ) MICE FINAL EFFEC- TIVE NUMBER OF MICE MAXIMUM INCIDENCE OF TUMORS (T.I./weeks) T A AVERAGE LATENT PERIOD RELATIVE FOR TUMOR CARCINOGENIC INDUCTION POTENCY, EL. in wksi PMC 33 C3H 2 100 20 11* 100/17 (12.71.7) ( ^ 2!:o^) oved A 33-1 C3H 2 100 20 16 94/17 3-5 C3H 1 100 3-5 CFW T 100 3.13a C3H 3 20 3.13b C3H 3 20 3-13c C3H 3 20 20 ' 12 20 19 15 12 15 12 15 13 92/22 79/28 0/30 0/30 69/30 3-13^ C3K 3 20 15 8 87/29 8-13 C3H 3 20 15 7 43/29 8-131* C3H 3 20 15 12 25/25 3-13g C3F. 3 20 15 9 100/27 8-I3I1 C3H 3 20 15 12 92/29 3-131 C3H 3 20 5 10 70/29 3-17 C3H 1 100 10 (20) 8 (17) 100/17 (94/30) 12-2 c?v: 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 23 93/59 L C3H 3 50 30 22 o/i+o i -- 2 Number of animals alive after 33 weeks Painting discontinued after 1+1 weeks. (1 2 .4+1 .6 ) (o.i*ot;gg) (I8.7 +I08) (2 1 .2+5 .6 ) - ~ 27.4 24,,1 - - (2 1 .0*2 .8) 21+,,7 ^ 25.3 (1 2 .4*2 .3 ) (39.0*5.6) (11.1*1.5) (34.6*5.0) m v 0.09 AJ0 .10 - - ^ 0,10 r-l 0.10 -eo-:i6 - -i 6-:S6 Oo93! ^ | 0.22+03 uo API 05672 T TABLE I ^ s k i :; p a i n t i n g on hic-:-: c o i l i n g s a m p l e s ^-- API _ ?u VUPCHF <2-k* TR.n lit CF MICE AND ITi-____i.rc CF ; s PER ..TEK <JA * - _L s CC O W H eu H <4 m o O M C CO fi 04 ORIGINAL O Pu NUMBER fi < CF (mg. ) MICE 100 10 (20) CLCC FINAL EFFEC TIVE NUMBER CF 5 T JitXUtu 10 (16) I 0 A L D ATA AVERAGE .IAa IKUM LATENT litwxLEitCE PERIOD RELATIVE 0F FOR TUMOR CARCINOGENIC TUMORS INDUCTION POTENCY, (Toi,/weeks) (5#FLoin wk$ Pmc 0/31+ (56/31+) (27.2+13.2) (0.09!;^) 12-5 cf v: 3 100 12-6 CF''.' 3 100 10 (20) 10 (20) 6 (1 2 ) 8 (1 2 ) 100/31+ (92/31+) 75/39 (7/33) (23.6+5.8) (32o2il2ol) o oe'1'0^ #u^-o01 API 05674 ) ) TAULE II CONCENTRATION STANDARDS WITH METHYLCHOLANTURENE IN BENZENE API SAMPLE NUMBER CONCEN TRATION (g./l00g.) DOSAGE PER APPLI CATION (mg.) STRAIN OP MICE AND NUMBER OP APPLICA TIONS PER WEEK DOSAGE oF ETHYL0H0LANTHRENE (mg./standard. area^/weelc) ORICINAL NUMBER OP MICE FINAL EFFECTIVE NUMBER OP MICE MAXIMUM INCIDENCE OF TUMORS (T. I./weeks) AVERAGE LATENT PERIOD FOR TUMOR INDUCTION STARTING 5% EL. in wks.) DATE 22 0 100 C3H 3 0 20 1 2 0/101 - 32 /4/50 201 0 .0 1 1 100 C3H 3 0.033 10 203 0.086 100 C3H 3 0 .2^8 10 9 44/71 7 2 .6*9 .2 7/20/49 ro O" 9 100/34 2 6 .2* 3.4 5/20/49 203 0.086 100 C3H 3 0.256 20 20 85/35 29.9*2 .2 10/0/50 2 19 # 0.114 100 C3H 3 0.342 30 28 96/35 24.0*2 .4 4/14/50 243* 0 .14 8 100 C3H 3 o.^l|4 20 2lj3 0 .l48 20 C3H 3 0.444 20 20 100/24 iy 100/26 17.9*2.0 18.2*2.1 7/ 2 8 / 5 1 10/ 1 / 5 1 204 0.171 100 C3H 3 0.^3 10 10 100/21 15.7*1.6 5/20/49 J . Area covered by 0.1 gram Benzene, 2. Number of animals alive after 30 weeks. i API 05675 ) ) TABLE II CONCENTRATION STANDARDS WITH METHYLCHOLANTHRENE IN BENZENE API SAMPLE NUMBER CONCEN TRATION (g./lOOg.) DOSAGE PER APPLI CATION (mg.) STRAIN OF MICE AND NUMBER OF APPLICA TIONS PER WEEK DOSAGE OF METHYLCHOL ANTHRENE img ./standard area1/week) FINAL ORIGINAI EFFECTIVE NUMBER NUMBER OF OF MICE MICE MAXIMUM AVERAGE INCIDENCE LATENT PERIOD OF FOR TUMOR TUMORS INDUCTION STARTING (T.I./weeks) (5 EL^ln wks.l DATE 20l+ 0.171 20 C3H 3 0.513 20 2 100/21 16 .6-1 ,2 9/26/52 20+ 0 .1 7 1 20l+ 0.171 20 C3H 3 20 C3H 3 O.513 O .513 20 20 100/21 l6.1+ti.i 9/26/52 rv> 20 20 100/21 1+.1-1.8 IO/28/52 -j 20$ 0 .31+2 100 C3H 3 1.026 10 10 100/11+ 11.0-1.9 5/20/1+9 20b 0.685 100 C3H 3 2.055 10 10 IOO/ 1 6 6.7-1+.2 7/20/1+9 P H8 0.228 20 C3H 2 0 .1+56 268 0.286 20 C3ii 2 0.572 20$ 0.3+2 100 C3H 2 0.681+ 1. I Area covered by 0.1 gram Benzene. 20 20 100/27 18.7*2.3 IO/2/51 20 19 IOO/2I (17.6*1.1) 2/13/53 20 20 100/20 15.1*1.5 6/9/50 API 05676 ) ) ) TABLE II CONCENTRATION STANDARDS WITH METHYLCHOLANTHRENE IN BENZENE API SAMPLE NUMBER CONCEN TRATION (g./lOOg.) DOSAGE PER APPLI CATION (mr.) STRAIN OP MICE DOSAGE AND OP NUMBER OP METHYLCHOL ORIGINAL APPLICA ANTHRENE NUMBER TIONS |mg./standard OP PER WEEK area^/week) MICE PINAL EFFECTIVE NUMBER OP MICE AVERAGE MAXIMUM LATENT INCIDENCE PERIOD OF FOR TUMOR TUMORS INDUCTION STARTING (T.I c/weeles) (5#EL. in wks .) DATE 227* 0 .1114- 100 C3H 1 O.llij. 20 15 07/1*9 I4.62.5 U/22/50 205* 209 0 .3il2 0 .5 14 100 C3H 1 20 C3H 1 0.3^ o.5iii- 1$ (2 0 ) 201 10 (19) 20 100/28 (100/35) 100/214- 2i.dt3 .l4, (22.1|2.7) l8.3ti.9 6/L3/50 10/1)/5 1 220 1.027 100 C3H 1 1.027 20 20 100/13 9.7*1.6 5/3/50 API 05677 1. Area covered by .1 ',i*ajn Benzene ) ) TAP. LE I I AJI SALILE i 'lm1CENTN A :.lON GTAn:A.cDS Vi/lTii MRTiIYLCU- .LALTRiiAiE IN BENZEl STRAF OF D SAJE .LA-IL MICE ARO or FILAI: NU1IEER 01' jFTHYlC.L-L 0i. ..1 AL EJFECT1'/E CORCEli- A. 1*'. -A- Al i .1CA- AlrPIL.idlE TLATiJ 1ir,t. no-is (mfc./sU; . .) .) HS . //2111area1/wk.) 1LI.1L EH *'O'F/1 T* tA Lb i;Uiotii OF :i ce- LAXILUI.; INCIDENCE OF TULORS (T.I./weeks) AVEHAuE LATENT PERIOD F jR TTILOR INDU CTl Oil (5F.E in w ELU 0 100 CFW 3 0 20 162 0/62 - STARTING DATE 12/1^/50 210 203 203 21 )a 2d! ! 0.023 0.08 A O.O86 J.lll]. J.I7 I ivi;/ 0?':/ 3 0 -o CF'.V 3 LOO CFW 3 ]00 crvv 3 1. t i r v i 2 i.O39 0.250 0.258 0.3l3 o .l v ; 10 (20) 10 10 (20) 10 (20) iO n (l/'l-'i 1 y 7 (1 6 ) io (1 6 ) O 06/U7 (79/53) 100/23 iOO/U (75/3D 100/20 (iOu/3 3 ) T O O 16 (,>5.9-11.3) 2/3/50 (17.1-3.5) 5/20/59 rv> vO (22.5-1; .3 ) 1-0/9/50 (I.I2 .3 ) 1 /15/50 (11.9-2.7) 5/2o/l|9 20 r J.3J|2 1.j-i or.; 3 1.026 10 20 9 . 3U2 LOO CF'.V 1 o.7ij: 19 10 i.00/2br 17 m o /30 u4.3* 5 . 1 (21.9*3.0) 3/2u/J.| 9 V 13/50 J'J'I .0 5 lu1- 1:\ i >v. ` 20 A m a .:.'"'ored by i.i 2. ,i h u r .'I a n i m l a al lei issile. o " f o I'-; u ^ e k o , 17 100/27 (l.i.5-2.5) '7i3/:.o API 05678 T \BLE II EXPERIMENTS INVOLVING THE APPLICATION OF SOLUTIONS OF METHYLCHGLANTHRENE IN SEC.-AMYLBENZENE TO THE SKIN OF MICE API CONCEN STRAIN SAMPLE TRATION OF NUMBER (g./lOOg.) MICE NUMBER OF applica7 TIONS1 PER WEEK ORIGINAL NUMBER OF MICE FINAL MAXIMUM EFFECTIVE INCIDENCE NUMBER OF OF TUMORS MICE (T.I./weeks) AVERAGE LATENT PERIOD FOR TUMOR INDUCTION (5# F.L. in wks.] RELATIVE CARCINOGENIC POTENCY, PMC 22*|.* 0 CFW 3 30 23 17/59 - - 223 0.087 CFW 3 223 0.087 C3H 3 10 (2 0 ) 20 (il) 16 89/^6 (83/I4.6 ) 81A 6 3 2 .3*1 0 .2 14-0.8^3-8 - Z f o l V*J o 0.05 .0 1 222 0.17*1- CFW 3 19 19 100/33 2 3 .6-2 .2 222 0.17*1- C3H 3 10 (2 0 ) 10 (2 0 ) 100/28 (100/1*4 ) 23.9*2.5 0 .10+' 02 " .01 API 05679 1. Dosage per application was 100 mg. TAliLK IX EXPERIMENTS INVOLVIN'! TIIK APPLICATION OP SOLUTIONS OP METHYLCHOLANTHRHNE IN VARIOUS SOLVENTS TO THE SKIN OP MICE I API IsAMPLE STARTING CARCIN NUMBER DATE OGEN1 SOLVENT a-- go 2 > WO h\ h0 3 n OO ^bo 1 STRAIN OP MICE AND NUMBER OF APPLICA TIONS PER WEEK DOSAGE PER APPLI CATION (mg. ) DRIGINAL NUMBER OF MICE AVERAGE FINAL LATENT EFFEC MAXIMUM PERIOD TIVE INCIDENCE FOR TUMOR NUMBER OF INDUCTION OF TUMORS (5% F.L. MICE (T.I./wks in weeks) RELATIVE CARCINO GENIC POTENCY, PMC I 210 2/3/50 MC API-52 0.306 C3H 3 100 20 17 100/30 (21) .9-1.1* o.ioi.oi 218 2/3/50 MC API-5 0.306 cpw 3 100 10 (20) 21*6 3/22/52 MC API 112-13 0.209 C3H 3 20 20 (1?) 20 09/25 (81/29) 100/22 (21.6-3.8) 1 5 .1* 1 .2 0 .08:;<a o .2o !;03 21*6 i*/n/52 MC API 112-1 0,209 C3H 3 20 20 18 100/21 15.0*1.1* o .2o !;3J 2l*0 l*/ll/52 MC API 110-13 0.201* C3H 3 20 281 11/12/5 . MC Cottonseed 0.01*1 C3H 3 100 oi I 20 38 20 100/22 (lo.7*1.2) 0.i8-+.*092i 37 62/69 1*9. o il* . I* o .o 3 .o i API 05680 1. 20-Methylcholanthrene (MC) . 2. Wilmington Residuum; see page 18 of Table I for description. 3. See page 19 of Table 1 ior description. ) ) ) T A U L E XI CONTROL EXPERIMENTS WITH VARIOUS SOLVENTS API SAMPLE NUMBER SOLVENT 80 95 221* 3 221J 22k* 228 228 2lj.l 2ifl White oil White oil Dodecylbenzene Dodecylbenzene Sec.-amylbenzene Benzene Benzene Distilled water Distilled water NUMBER OP APPLICA TIONS H- PEJJ WEEK 3 3 3 3 3 3 3 3 3 STRAIN OP MICE ORIGINAL NUMBER OF' MICE FINAL EFFECTIVE NUMBER OF MICE MAXIMUM INCIDENCE OF TUMORS (T.I./weeks) INCIDENCE OF CROSS' CARCINOMA STARTING (percent/weeks) DATE C3H 30 CPW 33 C3H 20 261 0/78 262 0/78 18 33/37 0/78 0/78 * 0/52 8/7/51 7/20/51 5/5/50 C3H 30 CFW 30 0 0/lk 23 17/59 o/ik 0/71 2/27/53 VjJ f\j 7/U|./5o C3H 20 CPW 20 C3H lio 181 0/101 162 0/62 301 0/78 0/101 0/62 0/78 12A /50 12/V50 7/28/51 CPW 17 122 0/70 0/70 7/28/51 1. Number of animals alive after 36 weeks. 2. Number of animals alive after 33 weeks. 3. Age of animals at start of experiment, 5k weeks. l|. Dosage per application was 100 mg. API 05681 SOLUTIONS CP SYNTHETIC C A P O I M O G E N S 1- IN DOEEG iLLENZENE* THE SKIN OP MICE API SAMPLE NUMBER CONCEN TRATION g./l00g.) STRAIN OF MICE NUMBER OF APPLICA TIONS PEN WEEK DOSAGE PER APPLICA TION (mg.) ORIGINAL NUMBER OF MICE FINAL EFFEC TIVE NUMBER OF MICE MAXIMUM AVERAGE INCIDENCE LATENT PERIOD RELATIVE OF FOR TUMOR CARCINOGENIC TUMORS INDUCTION POTENCY, (T.I./wksJ (5#F.L. in wksj fMC 221* 0 C3H 0 100 20 18 33/37 - 3 221 ^ o- C3H . J"> 100 \ 30 0 0/l/j - 226 0.006 CFW 3 100 20 17 1*1/51* - 226* 0.006 C3H 2 100 10 8 63/^38 35.2-11.1 (20) (12) (50A0) - - - V .> V) 225-::- 0.011 CFW 3 100 10 8 1*3.3*16.1 (20) (Ilf) <79%$ 225* 0.011 C3H 3" 100 213* O.Olj.6 CFW 3 . 100 211 0 . Lllp CFW 3 100 211 O.lll* C3H 3 100 208 0.ll*9 C3H 3 100 20 15 07/1*3* 3l*.oil*.o 1.061.01 20 15 100/18 10.5-1.7 0 ! r+ ? *~-.i9 8 7 100/17 11.5*2.9 0.33.t 20 20 100/22 1 0 .9*2 .0 - OC)*.n u -32-.07 30 30 100/15 10.3*.7 e TC'+.Of; -'^-.03 API 05682 1. The carcinogen was methylcholanthrene in every case except API-21J.5 in which case 3,l|.-bonzpyrene was used. 2. Product obtained by alkylation of benzene with the propylene-tetramer (aluminum chloride catalyst). . 3. Age of mice nt start of rjx eriment, 51* weeks. j. v., f'io'-i <^r nirror-.r i. < v iden of of Invasive malxgiv^ricy. - ) r.\iiLii. i n H | IMGNT3 INVOLVING TNC APPLICATION OF SOLUVLC.l OF CARCINOGENS* IN DCDECYLBEUZENE* TO THE SKIN OF MICE API CONCENSAMPLE TRATION NUMBER ig./lOOg.) STRAIN OF MICE NUMBER DOSAGE OF PER APFLICA- APPLICA- TI0N3 TION PER WEEK {mg.) ORIGINAL NUMBER OF MICE FINAL EFFEC- TIVE NUMBER OF MICE MAXIMUM AVERAGE INCIDENCE LATENT PERIOD RELATIVE OF FOR TUMOR CARCINOGENIC TUMORS INDUCTION POTENCY, (T.I./wks.) (5$F.L.in wks.] PMC 2^9 O.I72 C3H 3 20 20 18 89/34 10.lj.-i.lj. 2i|-9 0.172 C3H 3 100 13 (20) 13 (lb) 100/12 (100/1 2 ) 9 .1 -1 .2 0 . k^2 +- =0n7l 212 0.3*4 C3H 3 100 10 10 100/ 1I1 10 .2-2 .1 (20) (19) (100/1 8 ) -35- : i P M 1 0.172 C3H 3 100 20 18 100/19 11.3-2.0 21,5 1 0.172 C3H 3 20 15 13 100/31 17.0^3.7 214-51 0 .17 2 C3H 3 5 10 9 100/2i 16.1i2.7 nc) (11) (100/2 3 ) 5* ni n 895 - " -- .--- --O ---- J ~ 3,^-benzpyrene was used. --- j ---- -- r *- 2. Product obtained by alkylation of benzene with propylene tetramer (aluminum chloride catalyst) DESCRIPTION OF EXPERIMENT MATERIALS APPLII TO MICE DURING API EXPERI FIRST SIX WEEKS MENT OF NUMBER EXPERIMENT SEVENTH WEEK TO END OF EXPERIMENT STRAIN OF MICE NUMBER OF ORIGINAL \PPLICA- NUMBER TIONS6 OF PER WEEK MICE FINAL EFFEC TIVE NUMBER OF MICE AVERAGE MAXIMUM LATENT INCIDENCE PERIOD OF FOR TUMOR TUMORS % INDUCTION 3 (T.I./weeks j (5/FL in wks, 221-::9-::- DDB API-912 9* 221-9221-9 API-9 DDB DDB 253-9 ' PDD 57-::221-57* 208 221-208 a p i -572 *^ DDB API-208^ DDB DDB API-9 API-9 API-9 API-9 API-9 API-57 API-57 API-208 API-208 C3H 3 20 18 33/37 - CFW 3 io 10 lOO/lpO 28.l4. A a (20) (16) (100A D C3H 3 10 9 iooA 7 . 36.2A.5 C3H 3 20 20 100/21 14-0-2.1 VUAj C3H 3 34 11 100/22 A . 22 .6 (2 0 ) (lip) (100/22^ 6 C3H 3 7 100/20 1 2 .8A .8 (20) (17) (9/26) CFW 6 20 17 100/25 16.9*2.0 CFW 6 20 18 100/16 8.oil.9 C3H 3 30 30 100/15 10.3^.7 C3H 3 20 20 10 0 /llf 7 .2 -l.k API 05684 1. Dodecylbenzenes obtained by alkylation of benzene with 12-carbon olefins; DDB obtained with propylene tetramer; 2-phenyldodecane (PDD) obtained with 1-dodecene. 2. See Table I for description. i 3. Time measured from date of initial painting with carcinogenic material. 1|. Hair clipped. . ^ . See Table II for description. ' . Dosage per application was 100 mg. TABUS XIX EXPERIMENTS ON ACCELERATION OF CARCINOGENESIS BY SPECIFIC SOLVENTS API EXPERI MENT NUMBER DESCRIPTION OF EXPERIMENT MATERIALS APPLIED TO VIICE DURING FIRST SINGLE ? NINTH WEEK SIX WEEKS APPLICATION^ TO END OF IN SEVENTH OF XPERIMENT WEEK XPERIMENT STRAIN OF MICE NUMBER OF ORIGINAL APPLICA NUMBER TIONS8^ OF PER WEEK MICE FINAL EFFEC TIVE NUMBER OF MICE AVERAGE MAXIMUM LATENT INCIDENCE PERIOD OF FOR TUMOR TUMORS _ INDUCTION - (T.I./weeks / 5#FL in wks.J 229 Nil API-2293*^ Nil C3H - pO ko 12/13 - 23O * Nil API-229^ DDB C3H 3 lp 36 9 k /3 k 12.8-2.0 231 Nil API- 8 ^ 5 Nil C3H - 3k6 0 /b k - 232 Nil API-8^ DDB C3H 3 3^ 56/26 236 DDB API-8 DDB C3H 3 30 26 31/30 - - OOj' 237 DDB API-229 DDB C3H 3 ko 38 87/22 11.2^2.5 239 Nil API-8^ water C3H 3 30 216 0/79 - 2lp0 Nil API-8^ white oil, C3H 3 30 186 0/79 API-80 API 05685 1. Dodecylbenzenes obtained by alkylation of benzene with 12-carbon olefins; DDB obtained propylene tetramer; 2-phenyldodecane (PDD) obtained with 1-dodecene. 2 Single application followed by two week interval before further treatment of mice, 3, Solution of 0.5 g. 9 ,10-dimethy1-1,2-benzanthracene in 100 ml. sec.-amylbenzene. Ip. Hair clipped, 5. See Table I for description, 6. Number of animals alive after 63 weeks. 7. Time measured frcm date of initial painting with carcinogenic material. 8. Dosage per application was 100 mg. with API SAMPLE STARTING MBdJEi; DATE SOLVENT 2 !,), /iip/51 Benzene 9/1953 Naiizeif Ti' THE SKIN OP C3H MICE DOSAGE irOliEER OF op SOLUTION PER ORIGINAL CONCEN APPLICA TRATION TIONS (g./ iC":rui') PEL WEEK APPLICA NTOPER TION Op' (mf..) illCE O.iO ',Ji 100 7 (2 0 ) - 0.15 3 20 20 AVERAGE FINAL LATENT EFFEO- MAXIMUM PERIOD TJVE INCIDENCE FOR TUMOR NUMBER OF INDUCTION OF TUMORS [5% F.L. MICE (T.I./wksJ In weeks) RELATIVE CARCINO GENIC POTENCY, PMC 7 (2 0 ) 100/33 (100/3 3) 22.1*5.8 (25.1+2.5) n0 .1n2o_+.- 006^ o.io+.oi -.0 2 26)1. 12/19/52 APi-Ou1 0.15 3 261*. 12/1.9/52 a p i -Go o.i5 -*J1 J1/6/52 API-GO 0.2U 1 100 20 19 1 0 0 / 3 1 (25.5*1.3) 0 .1 0 .0 1 20 20 10 1 0 0 / 3 1 [25.k*i.5) 0U' J1`0U"+ .*02P 50 30 20 05A-1 /v/3 6 .0 'O 0.17 257 11/6/52 API-3- 2?; 0 .2 0 l (fraction^ of 2 .>6 1/16/53 frac tion ' o.i5 3 Df APT-G-1? diluted 50% with benzene 5o 20 16 83/kl ^ 35.7 ^ 0 . 1 7 100 15 13 62/30 ^ 2 3 . 7 0.08 API 05686 1. Teclmical white oil, vlscos ity <v!00 SSU/100, 2. Colorless fraction from chromt orraphy; see pages 21* and 22 for description. API SAMPLE STARTING DUMBER DATE SOLVENT STRAIN OF MICE AND NUMBER OF APPLICA TIONS TER WEEK DOSAGE OF SOLUTION1 PER ORIGINAL APPLICA NUMBER TION OF ' (mg.) MICE FINAL EFFEC TIVE NUMBER OF MICE AVERAGE LATENT MAXIMUM PERIOD INCIDENCE FOR TUMOR OF INDUCTION TUMORS 5% F.L. T .I ./wks.! In wee les) RELATIVE CARCINO GENIC POTENCY, PMG 270-1 2/13/53 Fraction b.520-600 C3H 3 (2 0 -5 0 )2 (1 2 -6).2 6 F./760 mm. from dis tillation of APIp80 10 0 /2 2 (16.63.1f) 2 7 6 -2 2/13/53 Fraction b.600-630 C 3N a (2 0 -5 0 ) 2 (1 2 -6 ) 2 6 F./760 mm. from dis tillation of Al'I-00 10 0 / 2 2 (I7.1f*if.5) o.20 j;*o 276-3 2/13/53 Fruction b.630-6lf5 C'3N 3 (30-50) 2 (1 2 -6 ) 2 5 F./760 mm. from dis tillation of APlr-80 10 0 / 2 2 (1 8 .i 3 .3 ) 2 7 6 -if 2/13/53 Fraction b.6lf5-655 C3H 3 (30-50) 2 (1 2 -6 ) 2 6 F./7 6 O mm. from dis tillation of API-00 67/26 a * 25.5 0 .0 9 27-S 2/13/53 Fraction b.655-700 C3H 3 100 15 F./7 6 O mm. from dis tillation of API-00 15 87/27 /\j22 .9 v 0 .11 API 05687 276-5 6/12/53 Silica gel chroma- C3H 3 50 3.5 15 0/10 tograpny of distil lation residue b. > 720F./760 mm. of APl-CO 1. Concentration of benzpyrene in solution was 0.15 g./lOO ml. solvent. 2. After If weeks the dosage was increased to 50 m g . and the numiaer of mice was reduced to 6 .i.i. l i t KXI'JSHXMii-N T S I N V . 'l . V i l l : . i'lil'J A i I 1.1 ' :AT J ON UK iiO L iU T lcniV , OK 1.liU 'A \'f\i liU K 1U \Ji\U 1 0 \lC . S U L V E U 'E J'O THE 3Kill OF C3H MICK API SAMPLE STARTING IUMBER DATE SOLVENT SATURATED FRACTIONS FROM CATALYTICALLY CRA(tOLD RESIDUUM NUMBER OF APPLICA TIONS i':5R WEEK DOSAGE OF . SOLUTION' PER ORIGINAL APPLICA NUMBER TION OF fog *).. MICE FINAL EFFEC TIVE NUMBER OF MICE AVERAGE LATENT MAXIMUM PERIOD INCIDENCE FOR TUMOR OF INDUCTION TUMORS (S o F .L . T.I./wks,: in wks.) RELATIVE CARCINO GENIC POTENCY, ... pwn 277-1 3/9/53 Fraction b .1|_30-500CF . 3 /760mm. from distil lation of colorless fraction of API-71- 30 0 ' 7 100/17 [13.5+2.2) 0 23+"07 277-3 3/9/53 Fraction b.55J_565cfc 3 20 6 6 83/I8 12.7-8.7 0 .26+ -?, /760mm. from distil -.14 u> -XD lation of colorless fraction of API-712 277--i|- 3/9/53 Fraction b.530-600F. 3 /760mm. from distil lation of colorless fraction of API-712 )|_0 8 8 88/19 A1 1 6 . 1 sv 0 .I8 277-5 3/9/53 Fraction b.600-620^, 3 /76(tani. from distil lation of colorlesg fraction of API-73. 50 0 0 75M rJ 20.5 0 .1 3 API 05688 1. Concentration of benzpyrene in solution was 0.15 g./100 ml. solvent. 2 . Colorless fraction of API-71 obtained by chromatography on silica gel. ) ) TAB L E 111 E X I '.HKIMENTS INVOLVING THE APPLICATION OP SOLUTIONS OF .ENZPYREHE IN VARIOUS SOLVENTS TO THE SKIN OP C3H MICE API SAMPLE STARTING DUMBER DATE SOLVENT PARAFFINS NTJMBER OF APPLICA TIONS PER WEEK DOSAGE OF SOLUTION1 PER ORIGINAL APPLICA NUMBER TION OF (mg.) MICE FINAL EPFEC- TIVE NUMBER OF MICE AVERAGE LATENT MAXIMUM PERIOD INCIDENCE FOR TUMOR OP INDUCTION TUMORS (5$ P.L. T.I./wks.' ln wks.) RELATIVE CARCINO GENIC POTENCY, PMC 269 3/9/53 Cetane (chromato 3 50 15 .> 1 1 36/10 -- -- graphed on si'lica gel) 273 7/2I1/53 n-decane 3 50 20 20 o /k - - CYCL0PARAFFI1JS 263 12/19/5* Cyclohexyldecane, 3 API-256 20 20 19 95/20 *0 1 3 .I ^ 0 .2 5 o 279 9/1953 7-cyclohexyltri- 3 decane, PSC-50^ ALKYLNAPliT HALENES 265 2/5/53 Dibutylmethyl- 3 naphthalene ko 8 20 15 11 100/22 (1 6 .3 2 .1) (20) (16 ) (100/25) 2/2 6/12/53 Monoamvlnaphthalenes , 3 liO 6 6 0/10 - - b. 570F./760 mm. P-73 6/12/53 Diamylnaphtlmlones, 3 50 9 7 Ii|/10 b.630-690F./760 mm. 1. Concentration of benzpyrene in solution was 0.15 g./lOO ml. solvent. API 05689 1 I XM `hlIi liN 11.V;11A'JN (! Tjlli Ai'l']_,1(JATHUl UF I'A I M .iKjOl1A1J1TloNU OF tiliNZPYKEULs, IK VAillOUA oOLVtii'.To TO THE SKIN OF C 3 II MICE API SAMPLE STARTING NUMBER DATE SOLVENT NUMBER OP APPLICA TIONS PER WEEK DOSAGE OF SOLUTION1 PER APPLICA TION (mR. ) ORIGINAL NUMBER OP MICE PINAL EFFEC TIVE NUMBER OP MICE MAXIMUM INCIDENCE OF TUMORS T.I ./wlcs. AVERA IE LATENT PERIOD FOR TUMOR INDUCTION ( 5 / f .l . in weeks! RELATIVE CARCINO GENIC POTENCY, PMC ALn tTL.JENZENES 275 6/12/53 Triisopropylbenzene 3 50 11 8 0/9 - ' - 262 ' /I9 /52 A1 -PpIh-2e5n5yIdocane, O 2l|.5 12/3/51 Dodecylbenzene P-hB 1 1 / 2 V 5 2 Dodecy1benzene 3 20 19 19 95/16 (9 .8 -1 .5 ) 0. 37! ; ^ 3 100 20 18 1 0 0 / 1 9 (11.3*2.0) I 3 20 15 13 100/31 (17.0*3.7) P- 2l|5 1 1 / 2 1 / 5 2 Dodecylbenzene 3 259 1 ;/2 1/ 5 2 20% Dodecylbenzene 3 in API-UO 260 l 1/21/52 5/6 Dodec; ibenzene 3 In API-BO 5 10 9 100/23 (l6.i|2.7) 0 ]7'*'o6 (15) (I D (100/23) x '-.03 100 15 13 1 0 0 / 3 2 (26.7*1.9 o u o r g i 100 15 15 100/33 2 0 .1 1 . 6 0 .0 8 1 . 0 1 API 05690 1. Concentration of benzpyrene in solution was 0,l5 g./lOO ml. solvent. 2 . 1'roduct from alkylai ion of benzene with propylene tetramer (aluminum chloride catalyst). API SAMPLE STARTING JMBER DATE SOLVENT TO THE SKIN OF C3H MICE NUMBER OF APPLICA TIONS PER WEEK DOSAGE OF SOLUTI ON1 PER ORIGINAL APPLICA NUMBER TION OF (mg.) MICE FINAL EFFEC TIVE NUMBER OF MICE AVERAGE LATENT MAXIMUM PERIOD INCIDENCE FOR TUMOR OF INDUCTION TUMORS (5% F.L. 'T.I./wks.] in wlcs.) RELATIVE CARCINO GENIC POTENCY, PMf! ALKYLBENZENES 27i|.-8 6/12/53 Fraction b.]j_50-500Ct 3 /760.B11. from -'iotil- lation of alkyl- benzene^ ko- 8 7 0/9 27JI--13 6/12/53 Fraction b.555-t>6Cf$\ 3 /760mm. from distil lation of alkyl- benzene 50 9 9 78/9 n i 0.6 'O 0.1*6 267-1 1/31/53 Fraction b.603-61*2$\ 3 /760ntri. from distil lation of alkvl- benzone(PDDB)3 20 15 11 91/27 ^ 22.5 sO 0.11 267-3 1/31/53 Fraction b.655-68 3 20 15 12 1*2/20 - - /760mm. from distil lation of PDDB API 05691 267-5 1/31/53 Fraction b.661*-73(ft1. 3 20 15 12 33/29 - - /760mm. from distil lation of PDDB j 267-8 1/31/53 Fraction b.659~76(ft'1 20 15 13 . i*6/28 - - /760mm. from distilJ 3 lation of PDDB I,. -- . . Concentration of benzpyrene in solution was 0.15 g./l00 ml. solvent. , Product of alkylation of benzene rrith propylene polymer. . Residual by-product of commercial alkylation of benzene with propylene tetramer. API SAMPLE HUMBER T A B l .k ; I X EXPiSRIMUNTd TO DKTEIMINE WHETHER CERTAIN MATERIALS HAVE IRRITATION AL PROPERTIES OP THE TYPE EVIDENCED BY DODECYLBENZENE DESCRIPTION OP EXPERIMENT IRRITATION LIKE . DODECYL BENZENE SEVERITY OP EARLY ACANTHOSIS AND IYPERKERATOSIS ---------------STRAIN OP MICE AND NUMBER OP APPLICA TIONS PER WEEK J DOSAGE PER ~ APPLICATION NUMBER CP MICE 221 Dodecy lboJiZene3 yes ' 221 Dodecylbenzene yes 2 1 - 1 1 1 -l\2% fraction obtained from chromatography yes ' ' ef Ji. 2 2 1 on alumina 221-2 i|.2-8i$ fraction obtained from chromatography yes of No. 221 on alumina 2 221-3 209 Doderylbenzene In API-UO no 221 -ij. l]0% Dodo-lbenzene in API-80 no 2 2 1-5 b0% Dodecylbenzene In API-00 yes 2 21-6 li0% Dodecylbenzene In API-80 . yes 253 2-plienyldor ecane yes 25)f. 3-plxeny ldodecane yes heavy heavy heavy heavy nil nil medium heavy medium slight C3H 3 C3H 3 C3H 3 100 20 100 301 20 3 -P- C3H 3 20 3 C3H 3 100 6 l C3H 3 100 7 C3H 3 100 7 C3H 3 ' 100 6 C3H 3 100 20 C3H 3 20 7 API 05692 1. Ape of mice at start of expex-iment, 5>ij-*week3. - 2. See page 32 of Table II for description. _) Fr.vluct from alkylation of benzene with propylene tetramor (aluminum chloride catalyst). EXPERIMENTS TO DETERMINE WHETHER CERTAIN MATERIALS HAVE IRRITATIONAL PROPERTIES OF TILE TYPE EVIDENCED BY DoDECYLBENZENE F DOSAGE PER " APPLICATION API SAMPLE NUMBER DESCRIPTION OF EXPERIMENT 255 256 280 71-3a 71-3b 1-phenyldecane 1-cyclohexyldecane e*-methylnaphthalene Material from cold trap of distillation of API-71-31 (0-2.5/i) Fraction b.l|65-530F./76o mm. from distil lation of API-71-32.5-6#) IRRITATION LIKE ' DODECYL BENZENE ' ) yea SEVERITY OF EARLY ACANTHOSIS AND HYPERKERATOSIS medium yes alight yea medium yea medium STRAIN OF MICE AND NUMBER OF APPLICA TIONS PER WEEK NUMBER OF MICE C3H 3 20 7 C3H 3 20 7 C3H 3 100 5 C3H 3 100 3 yea heavy C3H 3 100 3 71-ic Fraction b530-6lOF./76o mm. from distil yea lation offc API-71-3(6-ll^) 71-3d Fraction b.bl0-670F./760 mm. from distil no lation of API-71"3(11-10.7%) 71-3e Fraction b.670-690F./700 m m from distil no lation of API-71-3(l0.7-26?6) 221 |Dodecylbenzene yes medium nil nil heavy8 C3H 3 100 3 C3H 3 100 3 C3H 3 100 3 rabbit 3 300 2 Colorless fraction obtained by d-,r,'V"atrrraphy of API-71, TCG residuum. Severe crusting comparable to that in mice but occurring uetween the 9th and 15th week iupprox- A imately double the time required for the mice). API 05693 ) ) ) API SAMPLE NUMBER l i x j ri n Hi/jura t o TA1UJ! 1 I X d u t u h m i n k w u r -t h a u c u r t a i n .u a t k u i a l s iiavu i u r i t a 'I'i o n a u vkoturtiuu ,,, r ,;i -pYl'W EVIDENCED BY OOpZOYLUENTiENE DESCRIPTION OP EXPERIMENT IRRITATION LIKE D0DECYLBENZENE SEVERITY OF EARLY ACANTHOSIS AND HYPERKERATOSIS ST'.Ali: oi' ;..icz A*nD NUMBER OP APPLICA TIONS /s re .o-I i -T, i -i , 1 *mj i1. HUMBER n OF PER WEEK (n. .1 r.lICE ?76-L,2 Solutions of benzpyrene in fractions b. 520-' 30 of technical v/hl te oil 27b-J Solution of benzpyrene in fraction b,,6306^5 of technical white oil 3 7 6-[| 27 b-5 B7"-L 3J/j Solution of benzpyrene in fraction b c6lj.5055 "i technical white "11 Solution .-.f benzpyrene In fraction b,,655" 700 of technical white -..1.1 Solution of benzpyrene In fraction b e >720 of technical white oil Solution of benzpyrene in Cetane yes , s 1 ip.ht C3H ?. IB yes very slight CRT 3 30 IB 4= Ui no (scaling only) CJH 3 ;,0 IB no nil C3P 3 OvJ 15 no nil C3H 3 l.O i? yes heavy c in j 50 15 API 05694 ) API SAMPLE NUMBER TA.LK TV EXPEHIMENTS INVOLVIN':} A LIMITED NUMBER OF APPLICATIONS OF CARCINOGENIC MATERIALS ----------------- 1 STRAIN OP.MICE \ DESCRIPTION AND NUMBER DOSAGE OF PER ORIGINAL APPLICA APPLICA NUM3ER TIONS TION PER WEEK (mg.) OF MICE FINAL EFFEC TIVE NUMBER OP MICE /.VERAGE MAXIMUM LATENT INCIDENCE PERIOD OF FOR TUMOR TUMORS INDUCTION T.I./wks.) (9#F.L. In wks. n-1 1 Dewaxed oil from MEK-Btnzol C3H 2 20 20 19 0/6 - dewaxing of F.C.C. decanted oil No. U3 U3-12 , 3 U3-1 X 71 2 71 3 71 It n T.C.C. cracked residuum II n C3H 2 20 C3II 2 20 C3H 2 20 C3H 2 20 C3H 2 20 20 18 0/6 20 20 0/6 20 20 0/6 19 19 0/6 20 20 0/6 - - 4=" O' - - - 03-2 Aromatics bv chromatography from API-834 plus 30 >/! by weight of sec.-ainylbonzene C3II 1 100 13 (20) 12 (18 ) 83A95 (83/45) 3lj..6--t.Ij. 276-1, 0 .1 5 g. benzpyrene /100 ml. of C3H 3 ,3.A fractions of`ATT-00 b. 920 to j 695 ' _______________________________ 1 Painting continuf.il until last animal dies, a Painting of all animais discontinued at. time 3 Tainting of each animal discontinued at time 4 See page 2lj. for description, 5 Painting, discontinued after 19 treatments, 6 Painting discontinued after 11 treatments. 20 21 i 13 (6 each) 0/28 6 _____________________J first animal in group developed a papilloma. of appearance of papilloma In the Individual ' animal. i f' Ml API 05695 ') ) ) T ABUS V EX PER IM EN TS ON THE RETARDATION OF TUMOR FORM ATION BY W ASHING - CFW M IC E API EXPERI MENT 1 NUMBER 8 CARCIN OGENIC OIL APPLIED B1 NUMBER OF APPLICA TION# HAIR PER WEEK CLPPED * 1 no PERIOD OF EXPOSURE TO OIL BEFORE REMOVAL BY WASHING Control (no washing) WASHING AGENT none ORIGINAL NUMBER OF MICE 10 (2 0 ) 300 8 1 no 10 min. soap 9 solution ' > 30] 8 1 no 1 hour soap 10 solution 302* 8 1 no If hours soap 9 solution 12 121 3 yes Control none 15 (no washing) 303 12 3 yes 1 hour soap 15 solution 30lf* 12 3 yes 9 hours soap 15 solution FINAL EFFECTIVE NUMBER OF MICE MAXIMUM INCIDENCE OF TUMORS [T. I. /weeks) AVERAGE LATENT PERIOD FOR TUMOR INDUCTION :5#F.L. in wl^ 9 (13) . 0 7 100/26 (92/2 6 ) 0/81 llf/ 3 6 13.3-lf.6 -p-J - 8 63/70 (6 1 .62 6 .2 ) 1*4- 100/llf 1 0 .7-.9 lb 100/lf0 27.3-3.6 11 100/21 17.1-1.3 1. See Table I for description 2. Dosage per application was 100 mg. API 05696 4 ) ) ) TABLE V E X PE R IM E N T S ON T H E R E T A R D A T IO N OP TUMOR FORM ATION BY W A SH IN G - CFW M IC E API EXPERI MENT NUMBER NUMBER CARCIN OP OGENIC APPLICA- OIL t i o n s 3 HAIR APPLIEE PER WEEK CLIPPED PERIOD OP EXPOSURE TO OIL BEFORE REMOVAI BY WASHING WASHING AGENT ORIGINAL NUMBER OF MICE FINAL MAXIMUM AVERAGE EFFECTIVE INCIDENCE LATENT PERIOI NUMBER OP FOR TUMOR OF TUMORS INDUCTION MICE (T.I./weeks) (5#FJL. in wksj 305 81 3 306 8 3 307 8 3 308 8 3 113 1 1 3 1 3 309 113 3 310 113 3 311 113 3 yes 20 min. detergent 20 solution yes 1 hour detergent 20, solution yes 1 hour tiite oil- 20 detergent2 yes 20 min. white oil 20 yes Control none 20 (no washing) yes 1 hour detergent 19 solution yes ij. hours detergent 20 solution yes l;. hours shite oil- 20 detergent 2 Hi 93/51 U.'9*3.8 17 9ii/Mi 3li.O2.9 4- CD 20 85/33 26.62.3 15 100/52 31.9^.9 18 9 b / 36 (16,3i50) Hi ii3/67 - 13 5ii/57 - 12 83/52 k S . 3*k.6 1. See Table I for description 2. White oil rinse followed by washing with detergent solution, 3. Dosage per application was 100 mg. API 05697 ') ) ) EXl'ERIFEJiTS ON TIN1; iiETAlDATJwll OP TUMOR FORMATIO1 BY -WASH LH - C3-1 MICE NUMBER API CANC1N- OF EXPERI OGElilC API'LlCA MENT OIL TIONS1 HAIR NUMBER AiTLIEI PER WEEK CLIPPED PERloD OF EXP -SURE TO OIL BEFORE REMOVAL BY WASHING FASHING AGENT ORIGINAL NUMBER OF MICE AVERAGE FINAL MAXIMUM LATENT EFFECTIVE INCIDENCE FERIOD HUMBER OF FOR TUMOR or YUI.0HS INDUCTION MICE (T .I,/wee 1rs)(b/F.L.In wks) S6? 563 3 56 56 3 3lil 56 3 113 xo3 3 315 113 ** 3J6 none 3 m none 3 yes Control none 20 (no washing) yes Control (no washing) none 20 ' ye3 7 hours detergent 20 solution no Control none (no washing) 27 (1|0) yes 7 hours detergont 20 solution yes none detergent 20 solution yes none white oil- 20 ietorgent'-l- 20 0 /16 20 i.S/ 1 6 19 o/i6 27 (36) 20 1.00/2 3 (100/25) 0/16 20. o/i 20 o/:i6 - P" vO - 1 6 , it1.3 (1 7 5*13) -* - !.. Dosage- per application was 100 ,7ig. except where otherwise indicated, 2 Dosage per application was 20 mg. 3. See Table I for description. K- White oil rinse followed by washing with detergent solution. API 05698 ) ) ) TABLE V E X PGR TMEN ri 'S ON T H E 1i ET A RD A T 1 ON O F TUMOR FO R M A T IO N 13Y W A S H IN G - CFW M IC E API EXPERT- MENT NUMBER DESCRIPTION OP EXPERIMENT NUMBER OP ORIGINAL A P PLICA- NUMBER TI0NS1 HAIR OP PER WEEK CLIPPED MICE PINAL MAXIMUM EFFECTIVE INCIDENCE NUMBER OF OP TUMORS MICE (T.I./weeks) AVERAGE LATENT PERIOD FOR TUMOR INDUCTION C$EL. in wka) 312 Barrier cream applied 9:00 AM 3 API-8 applied 11:00 AM Removal of oil by by washing with detergent solution 12:00 AM yes 10 (20) 7 (10) 71AS (70A5) (1*2.3-6.0) 313 Barrier cream applied 9:00 AM 3 yes 20 API-0 applied 11:00 AM API-8 applied 2:00 PM Removal of oil by washing with detergent solution i+-:30 PM 13 100/31 214..6-2.7 O API 05699 I. Dosage per application was 100 mg, * memorandum RECEIPTS, EXPENDITURES, AND AlLCC-TIONS September 1, 1953 API MEDICAL 7S5: AP.CH EITTO /^nr.tributiona Received 1945 1947 1948 11995409 1199551? 1953 TOTAL $ 28, 900.00 27.300.00 98.300.00 64.340.00 94.840.00 109l7+., 426805..0000 79.056.00 $591+. 981.00 Expenditures 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 ^36,506.91 14,000.00 12.500.00 Philip Drinker Miscellaneous TOTAL EXPENDITURES BALANCE ON DEPOSIT SEPTEMBER 1, 1953 7 4 2 ,7 5 3 *7 3 463,006.91 15.300.00 ____ 512.17 521,572.31 $ 73,403.19 API 05700 9 P *=r E S S O L A B O R A T O R I E S STANDARD OIL DEVELOPMENT COMPANY P. O. BOX 51, LINDEN, .J. n|CAU RESEARCH DIVISION - E C K A R D T . PH D.-. M DD. ______ D IR E C T O R WG E R A R D E . PH D .. M O. HEAD TO XIC OLO G IST .C_V>C; 2 INDUSTRIAL HYGIENE SECTION N A T H A N V. H E N D R IC K S . CH .E . HEAO OF SECTION F R A N K L IN W . C H U R C H . M S. G EO R G E M. W IL K E N IN G . M .S . INDUSTRIAL H YG IENISTS -^trolsv.n Gonpsi*7 '/ills Oklahc-'na jr-ar Dr. Davis: ,s ncw ChalrriTR of the SPA Conaittee, I have prepared the attached report summariz irtngwtohueldabcetimviytiiensteonftitohnetoKeptrteesreinngt Laboratory as I see then this to the Subcommittee to on the present tire, Carcinogenicity feting in Houston as a forrjil report of cur RPA ConcitT.ee to the Sufccocxittee. Because of the unavailability of tins to cbtair. approval from each of the RPA jo-rdttee uenbers, I am unable to claim this as a full RPA Committee report. However. I plan to arrive ir. Houston at about two o`clock Ilonday afternoon, S ep tem b e r 26th. and have a reservation at the Shamrock Hotel beginning in the evening. I would be happy to go over this report with m y and/or all members of the RPA Committee that evening in order to introduce any corrections or sugges tions that night seem vrsrranted, or, if the Committee so decides, to abandon this r e p o r t altogether. For any members of the RPA Committee who will not be in Houston voaday 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 Comittee will hold that evening. Iiiff ahhn.o?shderin.sd3aingagrneyetshcroweniectehnctoospuiwreshiiocnfthehtrehpirafeetreaeltpsioornItosfthootuhDlred.hwaHovroekrteionfncutlihuetdheKdtehtietnetrshiuingsggeLrsaetbpiooorrnat,ttohraoytr, lie vd11 be in a position to advise me of the opinion of the Laboratory Konday svsm:ng in ITeuctcn. The two extra copies of the report being sent to Dr. Hcrton ere ior Dr. Kohoe and Dr. Phair.I 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 hsve btherent it represents, at least in ay mind, the important made thus far by the Kettering Laboratory, and in observations which the summary emphasizes the important points that should be pursued in the work of the Laboratory. It is h:pe that our RPA Committee. if not wishing to mcept this report, will deem it dvissble to prepare a report somewhat similar to this one which can be sub- itted to the Subcommittee cn Carcinogenesis at the Houaton meeting. If it is agreeable with thi3 Committee and with you, it would be ray plan to read this report API 05701 9 J *. (jgjaiLttee with the suggestion that it be forwarded tc tlie Safety Committee of tbe Board of Directors of the API. s i have indicated above, the report which I am sending to you now should be considered 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 Committee. j trust that our RPA Committee will be able to cos to some agreement on this report I'onday evening in Houston prior to the meeting of the Subcommittee on Carcinogenesis. Sincerely'; R. E. ECKARDT, I*. D. SEEsilk Attachment ~ with attachment! Dr* C. Hi-Hiae O tf E. K. Linder 9. Newqodet Mr. 5* J Adam* flr U O. -BWrci; Jlrv5 M M i * H. nsodaraca 'W z m H t M V (3) ) z ? K ' com mitids U M ir r a on ssp t b k b ir 29, 1953, TO U3C0MMITT5E ON CARCINCGIN2SIS, API diCAL ADVISORY COMMUTSS s"*3.---Y 0? TRY API PROJECT 3K CAP.CiyCGBNRSIS aT KSTTERUjG LABORATORY The original program of carcinogenic work at Kettering Laboratory grew out %f the r e s u lt s obtained from testing a series of 8 oils. The conclusion of t!l3 work, reported on November 5. 19^8, was that all 8 oils affected ths ekin. tv0 of them produced mild irritation, two produced a few verrucous papillomata, one produced papillomas and two cuestionable cancers, two produced a definite oncer each, and one wae 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 totting of indicated oils and tours, 3) chemical research to determine ths retare of the carcinogenic components end to develop analytical techniques for their determination, and, h) 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 safety 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 processes bm studied, in accordance with the outline on pages 2 and 3 of the lettering report dated October 27. 1930. API 05703 1 - 2- gnr.-acceieratinr: Solvants Ir. order to hs7o a rsi'erenc* 3ta-,dard tc which the results of tests of ths* oils could be referred, the laboratory undertook & study of the carcino genicity ?f solutions of ethyl cholanthrere in benzene. For reasons related to veil 'c.owr. dose-res :onss phenomena it wae decided to use the averrge latent oeriod for tumor production as a meaoure of carcinogenicity, the average latent period being defined as the length of time rec.uired oo reach 5056 tumor indices. In the coarse 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 standardise the mouse strain and to use only C3H mice in subsequent tssts, even though a considerable number of tests on oils with CFV mice had already been started or completed. In addition, it became apparent that if too strong a solution of methyl chelanihrene is painted on the C3H mice too frequently, the health of the mice, as determined by non-tumor mortality end weight carres, is impaired, end, apparently, their ebility to oroduce tumors is impaired at the same time. In adoition, other factors which influence the health of the mice, such as infections, infestations with lice, etc., are also reflected in non-tumor mortality end growth rates, and will seriously effect tumor production. Shoe unhealthy animals from any cause do not produce tumors as readily ae healthy animals, and the oreregs latent period is prolonged in the unhealthy animals. Since the aTerage latent period ie taken ae a measure of carcinogenicity, it ie 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 1 The f.C`r 3 c iE a rr/tic n s la? cT ,,hreo timee ct?r c: di lie 3iier.^th of s e c tio n beir-c: use--, me T however. h : ehovn that "a?e vhi'is carves c.ar c-e anttec. ir.` ;- a air*.'. curve *hen ih-ij* c.re expressed in ter.na of the dise of carcinogen applied per TMni; i-ea of skin per week. This has necessitated neaeurlng the area ci skin covered b7 a solution of aethyl cholanthrcne in a 6olver.t. A'lthir Units ft-s ' - ---- jnstant for each solvent tested. The solvents thus far ie, hydro-peroxide, free aayl benzene, cotton seed oil, 1 fractions of refinery streams. The effect of these ;t'on the spreading of the solution. Thus if the sail sentration of methyl cholaathrene in tvo different Lnted on the skin ? times a week, the dose per unit tae. If one solvent caused the solution to spread over ither solvent, the doss per unit area in the first cass he second case, and the average latent period for the ^iffe^rsnt, leading to different values for the two J^jSKfjpys pnl^ tp, concentration of methyl cholanthrene a yo^rence standard. What this work implies is tTtfyjf*3T work to determine not only the fra ction, and. the.amount applied per application, but also the area API 05705 1 Solvent wr.fti -i8 "`iS snid above apvlieB only to solutions of pure carclnogenB in ,,.r.-.-ceol^tins solvent. In the course of the wort: at the Kettering Laboratory .tt3 .-rterved that certain solvents, in addition to having an effect on the ..jo. :f --tr^Ed of the solution also have a specific effect on the carcinogenicity ,f the solution. Thus it was found that diffaran: concentrations of methyl .-olrntfcrese in some solvents, even though they spread over the same area, have ajFentiEllr the saB average latent period. According to the above, this should ,,}t 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, vjt the concentration varies, and hence does 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 this effect Is that the solvent exerts iose specific accelerating action on carcinogenesis. To summarise these two eoacepts, it may be well to give some equations. uy be defined at follow: 4. Thus dote per unit area per week where aga mgn of solution applied per application e * eoecentration U s *. V if .Pfllcatlo.. p.r ..dc covered nee-accslemtlag solvents with different spreading, Constant hat a varied, thereby influencing d. In the second a were constant but c varied and hence d also was variable. API 05706 I t,ut this did not influence average latent period. Hence two situations exist, ^ in which the rate of tumor formation is related directly! ) d, the other in ^jjich it is nt* To date dodecyl benzene and normal hexadecaae have been found t0 be accelerating solvents, although gasoline is not. An interesting observation recently made it that if dodecyl benzene is applied snce to the skin, and then serial biopsies taken, holes in the epithelium egg be observed microscopically but not grossly. It has been postulated by tbs Kettering Laboratory that these holes provide foci of rapidly regenerating spithelium which may be responsible for the accelerating action observed.*1 Incry Streams As a result of the above information of a fundamental n; tore developed over tbs past several years, tks fettering Laboratory now believes that carcinogenic refinery streams may ba divided into two main classes: 1). Those streams or materials in which PMC is related to d. 2). These streams or materials in which F^g la net related to d, In the first group fall' those materials such as catalytically cracked residua in which carcinogen.concentration K'tfd"high or aecslsratora relatively io low that the carela^m* ccnasalnmfiou becomes of first importance as a measure of tumor activity thauamamml groan, characterised by medium distillates. the accelerator concentration Important, in producing tuaors bam developed information which indicates that the accelerators tfs present in those, refinery streams boiling from 450-650*?. * while carcinogens trs present in those streams boiling above 650-700*?. Thns in mixtures which API 05707 I a vide toiling range, say from ^50-1000*1., the carcinogenicity of the pasture nay not he dependent alone on the amount of 700F. + material present, ,,at 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 in Pittsburgh, and have relt ted the determined benzpyrene content to the PMC (appendix II of January - September, 1953, Progress Report). these dtterminations indicate that the % BP/P^g ratio essentially divides into two group 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 (AFl^, 71 and 91) this ratio lay between 0.1 and 0.3, and these three oils aay arrant further study. table* of PMC In the newly distributed tablet o f Pjg. values distributed on September 1, 1953. issentially three types o f result are givens 1). Those in vhidifboth the average litent period end the PMC values v s. do W naei|jnall s variable ia the biological experiments such m poor toxioity of applied matrials, etci p S the valoo to not. Those are pretty good experiments from a biological otandpoint, but tho laboratory still has some reservations about the accuracy. API 05708 3). Thoee la which neither the avert-ge latent period nor the value are enclosed in parentheses These experiments 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 fall 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 highly desirable that these PKC values be made ae accurate as biological experimentation will permit. Another serious defect of these tables is that in some experiment*, anlhals 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. TIT is to he desired that no results will be reported unless a minimum numibeif'of 2C animal* are included in the experiment. The Committee recognise* the validity of throwing out some animals, but feels that where this has least 20 animals't.hin S&neAh* experiment should be repeated to include at . Other Ob Tha ilki fe&isg is an effective way of reducing the care iMj^ah cd^Vb tha skin of tha mouse. In general, the sooner the oil wHftd'off the better. Limited experiments with white oil wash and barrier cream* are difficult to interpret, but the laboratory feels that a white nil rinse prior to washing, or the use of barrier creams in association with washing, may offer advantages greater than with washing alone. API 05709 3 3 I 5fce epidemiological data ie accumulating in the laboratory, with some 1200 ^ge now haring been reported. No definite trends are as yet discernible but ^ was not anticipated th:t any such trends would be discernible prior to at 1<aSt 5 7ars experience. It is hoped that the supoort which the General g^it.tee, Division of Refining has given to the epidemiological study will jtisulate renewed reporting of cases with sufficient medical and occupational biitories to permit sole early correlations to be made about 1956. gsssa 1. The RPA Committee believes that the Kettering Laboratory is performing uportont work in the field of carcinogenesis of refinery streams and materials. Qiey believe that although a great many practical answers hare already been obtained, the work should be pursued because the work of the laboratory promises ,teprovide many fundamental answers to questions which now exist concerning trial and occupational caroinoganeaia. 2. It is believed that the work with noiwacceIsrating solvents which relates the carcinogenic reepottSe'of a eolation of pore carcinogen to the doee applied per unit area par vaeS^W'tltt ekin la of extreme fundamental importance. The Committee urges W `conducted to establlah firmly the curve t a*-* b W V c f i c with accelerating solvents le also of :*(8af this should be pursued vigorously in eosquste understanding of the relative role of carcinogens m ihTreflnery streams and materials. , a- API 05710 - 0 U, Since much of the correlation of chemical analyses with biological -^tency i* pendent upon the accuracy of PM{, values, the Committee Ljnento the fiCt thct nany of the -valueb thUB far reported are based on data which tbe ls'ccratory recognises as not having been obtained from the beBt biological eXperlScnt possible. The Committee suggests, therefore, that tha laboratory gvote considerable effort to repeating those experiments in an effort to develop ?MC valueB that are based, on the soundest possible biological experiment. 5 . The Committee believes that the work with washing techniques and protective cream* may nrovide intensely practical guides to refinery hygiena practice and suggests that this work be pursued as new approaches appear to the laboratory. 6. The Cosmittee recognises that the epidemiological studies cannot be expected to produce significant results until at least 1956, and, therefore*, urges that renewed efforts at adequate case and occupational history reporting U mads 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 Coamiittee, Division of Befining has given to the attainment of this objective. 7. The laboratory has already determined that a number of refinery streams . ..let *-i . . . . sad materials have a' sr-ve*lat<i&vely high dagree ter ji of . carcinogenicity. . . Although data obtained with be directly transposed to humans, there is sufficient data in' ** if alti,lomvavl:cancer experience to indicate that the pstroleomifipttsjy csnno$ take lightly the observations of the laboratory on the carclmegnleity of these refinery streams and materials. 8. The Committee reoognises that the laboratory at beat can determine from its biological and chemical work only the relative carcinoganleity of various refinery etreams and materials. Tha determination of the hasard is API 05711 ? 10 -- ^apend8r.t upon factors of extent, frequency, duration, and length of exposure. w reconlzed *>7 the laboratory in Dr. Kehoe5e talk to the members of the .>6?b1 Committee, Division of Refining the determination of the hazard ie at industrial hygiene staffs are essential in these indiTidu&l companies if they 870 adequately to appraise the hazard. The Committee supports Dr. Kehoe1s Dla that petroleum company managements recognize this truism and make plans for the development of each adequate medical and industrial hygiene staffs. 9. It is apparent that the laboratory has 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, and urges that the mechanism be established so that these publications can be reviewed in a finite period of time. As a suggestion, it is proposed that this mechanism be so established that review by API ssatgemsnt*and digestions end criticisms for the laboratory froa API management^ within sixty (60) days. If no criticisms or suggestions are attbs expiration of this time, the laboratory cAM forthcoming and proceed with publication. Itshould be rsc^i^^'^^pajl^^rl^^^0r the'purpose of helpful criticisms elK or not there will be ioryelearly states that the r expose:*-, * < API 05712 r1 SUMMARY of I ) recent progress in the investigation of the carcinogenicity OF p e t r o l e u m i n t e r m e d i a t e s a n d p r o d u c t s w 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^, since these are based upon experiments with such solutions as external standards of reference. Other materials from refining operations have exhibited potencies, Pjgg, 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, PjjC, with variations in conditions of exposure, simply on the basis of the unit from *hlch 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 as to render it insoluble, the experience of the succeeding n0nths has already shown that such is not the case. There is a fflounting 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 1 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 nolecular 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 i;-ringed polycyclic aromatic hydrocarbons. API 05714 - 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 80 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), l;. the relative potency, PMq , 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, methylcholanthrene in benzene), may be expressed algebraically by an equation of the following type: PMC * / (cd) !>rea] [KBPCBP `W 'DA Kc`] 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, Pjjq, is independent of the severity API 05715 I - k- f exposure and depends only upon the dosage of carcinogens |t-ne summation in brackets of the effective concentrations of benzpyrene, "Diels-Alder" carcinogens, and the third class, as y0t undefined) per unit area of the skin. The relationships between concentration of carcinogen (in g . / 1 0 0 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, (x pap - 2 . 8 ) (d + 0 .0 9 ) = 8 .5 ( 1) For the average rate of induction of grossly malignant tumors, (x ca - 1 5 . 5 ) (d + 0 .0 7 ) = 6.3 (2) (3) where f z number of applications per week. The relative areas of the akin 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, ao long as the solvent is a non-accelerator, i.e., sec.-amylbenzene API 05716 T I - 5 - oT cottonseed oil. For example, a given weight of a solution of jnethylcholanthrene in sec.-araylbenzene covers 1,7 times the area | c0Vered by the same weight of a solution of raethylcholanthrene in benzene; 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, PM(, 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 the rate of induction of tumors by polycyclic carcinogens. Ap l 05717 Doaage (tag, of Methylcholanthrene/Standard Area*/Week) ) EXPERIMENTS INVOLVINO THE APPLICATION OF SOLUTIONS OF HCETHILGHOLANTHHENE IN UPON THE SKIN OP C3H MICE I APPENDIX I ANALYSIS FOR BENZO(a)PYRENE IN REFINERY STREAMS (1) Prepare a chromatography tube by joining a 300 ml. reservoir 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 mm. 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 oil to be analyzed (added to column as a solution in 10 ml. of 50:50 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 redissolve 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 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 tqueous sodium thiosulfate. API 05719 APPENDIX I (Continued) jjj_) Chromatograph B similarly, without the use of iodine. Ijj) Equalize the weights of A and B (to W grams) and determine {110 spectrum (365-1+15 myt. ) 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 (6) Draw a straight line intersecting the absorbance curve at my and 380 m y . Let the difference in absorbance between the straight line and the curve at 39 he Z^Dgp. Then, % BP s A g B E . x -- i -- a ^ DBf M L 35 .87 3 0 .5 tile it is obviously not necessary to determine the entire spectrum (difference in absorption between the iodinated A and the jcn-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 my/ and the jositive peak of 6-iodobenzo(a)pyrene at 1+05 my/ T type OP STREAM catalytically packed Residuum API SAMPLE NUMBER 8 9 12 20 26 43-1 48 50 63 71 82 APPENDIX II PERCENT BENZPYRENE (BP) 0 .3 4 0,07 - 0.09 0.42 0.12 0.63 o.47 0.32 - 0.36 0,07 - 0.08 0.05 0.10 0.53 v0n 1 o . CD Pup (100 m g . ) 0.06 0. 42- 0,46 0.16 0 .6 5 0.1*3 0.22 0 .1 5 0. 14- 0.19 0.55 0.33 % BI ~^MC 0.5 1.33 0.95 0.75 0.96 1.09 1.55 0.50 0.30 0.18 1.61 6 Thermally Cracked Residuum 65 U3 Medium Distillate 2 3 28 33 79 91 102 105 0.02 0.06 0.09 - 0.10 0.02 - 0.03 0.02 0.01 <. 0.01 0.03 - 0, 01*. 0.01 0.01 0 .0 8 0,12 0.18 0.25 0.31 0.51 0.1; 0.28 0.19 0.11 0 .2 5 0.50 0.53 0.10 0.06 0.02 < 0.07 0.13 0.05 < 0 .0 9 ?avy Cracked 41 ^stillate 1.03 0 .3 4 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 Marion Duvall Richard Graeschel Lenore Hull James Pancake Jean Rehm Elizabeth Rush Effie West Report: A. Wesley Horton, fch.D Approved: -- ^1 Robert A. Kehoe, M.D. Director Sate: September 29. 1953 y.wo 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 f l n n m v i u n h r M o n September 29, 1953 to the Subioftunittee-on Carcinogenicity- of the API Medical Advisory Commit* tee. By verbal arrangement with 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. E. ECKARDT, M. D. REE:ilk Enclosure API 05723 REPORT OF RP (MC-1) ADVISORY COMMITTEE SUBMITTED ON SEPTEMBER 29, 1953, TO SUBCOMMITTEE ON CARCINOGENICITY. API MEDICAL ADVISORY COMMITTEE fltlMMARY OF THE API PROJECT ON CARCINOGENICITY AT KETTERING LABORATORY The original program of carcinogenic work at Kettering Laboratory gj-ew out of the results obtained from testing a series of 8 oils. The con clusion of this work, reported on November 5, 19L8, 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 U) an epidemiological program to develop Bound information regarding the actual incidence of cancer among employees of the petroleum industry and to determine the efficacy over a period of yearB 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 oethylcholanthrene is painted on the C3H mice too frequently, the health of the mice, as determined by non-tumor mortality and weight curves, is impaired, and, 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 Miiaals 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 that the apparent carcinogenicity of a meterial could be profoundly influenced ty 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 doee 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 t What has been said above applies only to solutions of pure earctasgens 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 some 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 ease 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 ease. 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 = mgm 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, but a varied, thereby influencing d. m 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 erectly 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 of 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 Fjg 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 650-700P. Thus in mixtures which have a wide boiling range, say from k50-1000F., 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 l+50-650F. 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 (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 * p.yc In the newly dlstrlhuted 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 PMC 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 Pj^ values 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 critically 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 P,_ values, it is highly desirable that these. ?HC value*,be made as SCiurate.es.biological experimentation vUl 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 fev 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