Document VGN1Be2DXO3XdXm5L33LKBppo

TITS INDUSTRIES ykTo: Messrs. A. G. DiMarco A. T letzsch INTER-OFFICE CORRESPONDENCE Date: February 14, 1974 From: Zeb G. Bel3, vJr. Location: 19 East Subject: NIOSH Consultants' Meeting in Cleveland, February 12, 1974 I am enclosing a copy of Dow Chemical's Health Inventory Examina tion, which is followed by them in obtaining a history of employees who worked with vinyl chloride. You may wish to review the attached for comparison with our medical history questionnaire. At the NIOSH consultants' meeting held in Cleveland, Ohio, on February 12, 1974, there was some preliminary discussion on a medical examination protocol for workers manufacturing PVC. This is not necessarily a concensus of the group who had assembled, but it does reflect their preliminary thinking. 1) A physical examination and history to elicit information that might be helpful in identifying abnormalities that may suggest reasons for exempting certain individuals from working with chlorinated organics was proposed. 2) For poly cleaners, X-rays of the hands have been suggested. 3) The following tests were suggested: a) b) c) /V 0 d) Chest X-rays of the individual Pulmonary function tests (FEV-1 and total) Complete blood count (CBC) Platelet count 4) The SMA screening tests were discussed for liver function. Those thought of some value were: ^ a) (/b) \,U \j c) SGOT SGPT LDH t/tf) Alkaline Phosphatase , e) Bilirubin 5) It was suggested that a prothrombin test may be of some value as an indication of an abnormal liver. FORM 503-4 Rev, SL 027248 Messrs. DeMarco and Raetzsch February 14, 1974 Page 2 6) Some discussion was held concerning the alpha fetal protein. 7) It was brought to the group's attention that the publication in the January, 1974, AIHA Journal of "Serum Ornithine Carbamyl Transferase as a Liver Response Test for Exposure to Organic. Solvents" should be reviewed for application to PVC workers. (See attachment) In conclusion, NIOSH and its advisory group are attempting to evaluate more definitive diagnostic tests for the early detection of liver involvement and are beginning to focus on other areas, such as the lung. Attachments cc: W. R. Harris R. E. Widing L. B. Grant, M.D. H. B. Love-joy, M.D, L. F. Sargert SL 0272** ' I X Af-' I A I t Mi Die A< i:>,AM UA M r'i PAH I Ml IIT NA ML HCjMF Aliniil'-iA Tbuilding n imber HEALTH INVENTORY EXAMINATION master no. (AGE IBlRTHOll E JOB CLASSIFICATION START DATC-PEPT I- HOMI I'HONI i: A"l V(*M f91C fAN'i^ AOOReS i ' DO YOU WANT RESULTS OF THISS _____Q DATE i(UI YES SCREENING EXAM SENT TO YOUR P'HHYYSSICIAN [J NO YEAR OF YOUR LAST DOW PHYSICAL EXAM. OR HEALTH INVENTORY --------- SMOKING HISTORY FAMILY HISTORY __ _ ITEM Dill YOU EVER SMOKF CIGARETTES Hat any blood rotation (grandparent!, parents, brothers, sisi children, aunts, uncles, cousins) had: DO YOU SMOKF rIG ARE T TES NOW______________ ITEM HOW MANY cigareites per day DIABETES HOW MANY YEARS(totolf HAVE YOU SMOKED CIGARETTES IF YOU HAVE QUIT.HOW MANY YEARS AGO 00 YOU SMOKE A PIPE OR CIGARS Do you loot that exposure to plant chemicals has had any elfeci on your health in the post year. ["H Yes [~1 No IT YES, WHY: HEART TROUBLE CANCER STROKE GLAUCOMA TUBERCULOSIS ABNORMALITY AT BIRTH HOW MANY CHILDREN DO YOU HAVE HAS YOUR WIFE HAD ANY MISCARRIAGES DID ANY OF YOUR CHILDREN HAVE AN ABNORMALI TY AT BIRT H I Wi RE ANY ol YOUR CHILDREN STILLBORN o. DIE WITHIN 6 MO'S of RiRUI 1 SINCE YOUR LAST DOW PHYSICAL EXAM OR HEALTH INVENTORY, HAVE YOU ITEM T6S NO - ------------------------ rrm------------------------------- rtfi SERIOUS ILLNESS OR INJURY 12 EARS, NOSE OR THROAT TROUBLE 27 HOSPIT AI.I7AT ION major opcraiion n REACTIONS to DRUGS or CHEMICALS 2B H ALLERGIES 29 ( HANOI- IN ENERGY 1 EVbl IS WHEEZING IN CHEST 30 CHALK.1 IN I'A UNO HAIU1S 16 COUGH LASTING OVER 3 MONTHS 31 ! XCC-SS 1 HIRST 17 COUGH UP BLOOD 32 CHANGE IN WEIGHT IB NIGHT SWEATS 33 CHANGE IN SLEEPING HABITS 19 SHORT of BREATH WHEN WALKING 34 OF PRES ION 20 COUGH AT NIGHT 35 EXCESSIVE NERVOUSNESS LOSS OE MEMORY r ITS, CONVULSIONS or SEIZURES ?l 22 23 SWOLLEN ANKLES 36 CHEST DISfOMFORT OR COID WEATHER WITH EXERCISE 37 NUMBNESS IN HANDS OR FEET 38 RECURRING severe HEADACHES 24 DIFFICULTY IN WALKING 39 D!Z7'G'SS OR FAINTING SPELLS 25 BLOODY NOSE 40 b URRrD OR DOUBLE VISION 26 EASY BRUISING 41 H--A--D----A-N--Y----O--F---TrrHTTE1--F--O--L-L-O--W---IN--G------ FREQUENT INDIGESTION or HEARTBURN 42 INTESTINAL TROUBLE 43 YELLOW JAUNDICE 44 BONE OR .JOINT PROBLEMS BACK TROUBLE 4b At- SKIN TROUBLE ALBUMIN or SUGAR in URINE PROBLEMS WITH URINATION CHANGE in URINARY HABITS a; 48 49 50 SEE HALOS USE ALCOHOL & DRUGS in EXCESS CHANGE IN BOWEL HABITS BLOOD IN STOOLS TARRY STOOLS 51 57 f\ 5J 55 OTHER: 56 COMMENTS: r HAS A PHYSICIAN EVER ' 'f M ASTHMA ADVISED YOU THAT YOU HAVE OR , f* N " ITEM 57 PROSTATE PROBLEMS HAVE HEAR r TROUBLt 58 ARTHRITIS HIGH 1 'LOOD PRESSURE 59 CANCER or MALIGNANT GROWTH t.IVFR TROUBl E 60 X-RAY proven ULCER |K 1 CNfc Y ur BLADDER TROUBLE 61 X-RAY PROVEN GALL STONES COMMENTS. HAD: 7E, N C ITEM 62 TUBERCULOSIS -- ' f- $ 1 t 67 63 ANEMIA or BLOOD PROBLEMS 68 64 DIABETIC CONDITION or TENDENCY 69 65 OTHER: 66 H70 ___ 027250 SL MAS 1 EM NUMBF K I Nut A fJUMMl M FILM NO. X-RAY 1 <.G. ID ? 2D 3 3 EXPECTED r vi VITAL CAPACITY OBSERVED rvc FF V T IMF FOR I VC VISION GOTH =u<r) RIGHT LTFT I FAR i 2. NEAR *,tt I ilC A 1 IOM mi inr. l a k i m M t <.i ' t CHEMISTRY CODE '.L UCOSC-RANDOM 38 5 BUN URIC ACID CREATININI CALCIUM 3:1 325 370 030 I1IURUB-T OTAL 315 PROTEIN: TOTAL 352 ALBUMIN 350 r.GOT 395 PHOSPHATASf-ALK 341 LDH CHOLESTEROL 311 045 SllFil.lir.n1 Ui ll ------- --- blood " SYS pressure IJIAS ___________ RESULTS URINALYSIS CODE 4 M r i ii ic *,PA n \ Y 750 4 pH 755 3 PROT F IN 710 3 SUGAR J KETONL 715 800 3 CCCIJLT BLOOD 801 3 MICRO 3 LEUKOCYTES 720 4 ERYTHOCYTES 721 4 CASTS 722 5 L rJTMF UAI CEU j 723 5 SYS DIAS. RESULTS DFPT H intraocular TENSION REPEAT HEMATOLOGY 2 RBC 3 HEMOGLOBIN 5 HEMATOCRIT 5 WBC .1 NEUT. 3 STABS LYMPH 5 MONO 5 EOS 5 BAS 5 ATYP.LYMPH JUVENILES CODE 420 410 415 425 430 433 431 4 32 4 34 435 436 437 RESULTS ! I r 9 4 4 6 4 3 4 -i--..............., 3 3 11 3 3 4^. ___ _ ,, .. = _. rSICIAN'S EXAM. NORM. ABNORM. CHI CMOFACH ITFM OR WRItr N.I . i Upt 1 Mil.) 1. LN I NORM. ABNORM. CHFCKF7 )EACH ITEM OR WRITE "N.E.''Not Eval.) 11. HANDS 7 1 VS- GENERAL 3, F UNPUSLOPIC 12- UPPER t X TREMI M ES f s trr-n^th, rnncje of mot t on) 13 FEF1 4 LUNGS AND CHEST 5. HEAR! (thrust, rhythm, sp""ri*) 6 VASCULAR SYSTEM (Varicosities 'Me*/ 7. ABDOMEN AND VISCERA uncludr- Herntn* 8 ANUS AND RECTIJm(p) 14 LOWER EXTREMITIES 15. SPINE, OTHER MUSCULOSKELETAL 16 IDENTIFYING BODY MARKS, SCARS, TATOOS 17 SKIN LYMPHATICS 18 NEUROLOGIC 9 ENDOCRINF SYSTFM 10 G U SYSTEM 19 PSYCHIATRIC (specify any personality devigtion' JHY$ICIAN'S UMMARY: tFc - -I r, i i i t I/ AAn Iy'H T 1 141 I- < fs I or; c p-| * l I A T A N-M *. i\ l I AH p*l H fc I f m ' LJ V - f* / y FINDINGS OF OCCUPATIONAL INTEREST D" ` D FOLLOW UP REQUIRED \ / A Ml ri Nt M.U. (-pure IL L--J Af-f r. with D RECORDS D i I'- A Ml 1 y I'HYSKIAN SL 027251 p boss. Sleep; ! 50: MOL: tk-Jiesi i vation. Report r 5, 1973 Scrum Ornilhinc Carbamyl Transferase as a Liver Response TesL for Exposure to Organic Solvents G. D. DIVINCENZO, Ph.D., and W. J. KRASAVAGE Ih'iillh wnl Safety Laboratory, liastmati Kodak Company, Rochester, blew York 14650 Ornithine larlnnnyl lran.sFerti.se (OCT), an enzyme found predominantly in the liver, is released into the bloodstream when liver cells are ruptured. The measurement of scrum OCT activity is a convenient, specific, and sensitive assay of liver damage. This, test was used to evaluate the effect of several widely used solvents on the livers of guinea pigs. Each solvent was administered intraperitoneally, and 24 hours later serum OCT activity was measured. Many of the solvents tested failed to increase serum OCT activity even at near-lethal doses. Of the thirty-three solvents evaluated, two produced elevations in scrum OC'l' activity at relatively low doses (less than 50 mg/kg), five at moderate doses (50 to 500 mg/kg), and nine at high doses (greater than 500 mg/kg). Introduction ORNITHINE CARBAMYL TRANS FERASE (OCT), a mitochondrial en zyme, catalyzes the condensation of L-ornithinc and carbamyl phosphate to L-citrullin.c and inorganic phosphate.' This conversion lakes place in the urea cycle of the liver and is a step in the cyclic process which detoxifies ammonia. In mammals OCT is found almost exclusively in the liver where its predomi nance forms the basis of its utility in detect ing liver damage.- '* Advantages of the OCT assay over other clinical liver enzymic tests include case of performance, true organ specificity, and enhanced sensitivity. In nor mal serum the level of OCT activity is lower than the activity of several other common diagnostic enzymes. Consequently an in crease in enzymic activity due to incipient liver damage may be more easily detected by the OCT assay. There is frequently a need in occupational health surveillance to establish with greater certainty the extent of employee exposure to organic solvents. Tests that measure organ function may be useful in this regard, but they frequently lack specificity, as for ex ample serum glutamic oxaloacetic transa minase (SGOT). The sensitivity and speci ficity of the OCT assay as a measure of liver response suggests that it may have use in biological monitoring particularly in test ing organic solvents for acute hepatotoxicity. A variety of organic solvents was tested in guinea pigs in a study of the correlation be tween serum OCT activity and histopathological liver changes. The results of this study are reported in this manuscript. Methods Each solvent tested was injected intra peritoneally into mature naive male guinea pigs. Prior to injection, the solvent was dissolved in corn oil, or, if the volume neces sary to deliver the calculated dose was too large, it was injected undiluted. Animals were kept separately and fed ad libitum Purina lab chow and water fortified with ascorbic acid. Fresh lettuce was provided twice weekly. Twenty-four hours after the solvent was injected, at least 2 ml of blood was withdrawn by heart puncture with the animal under ether anesthesia. The blood was centrifuged, and the serum was removed and immediately frozen until the time of SL 027252 O') analysis. OCT is stable at -- 15C for up to one year without any loss of activity.1 He molysis does not interfere with the deter mination, since OCT is not found in red blood cells.4 After the blood was collected, each guinea pig was sacrificed with an overdose of diethyl ether, the liver was removed, and pieces were fixed in buffered 10% formalin. Some of these were embedded in paraffin, sectioned at 7 microns, and stained with hematoxylin and cosin for routine microscopic examination. Other pieces of tissue were fixed, frozen, and stained with Oil Red O to detect lipid deposition. Serum OCT activity was measured on a Technicon Autoanalyzer by the automated spectrophotomctric procedure of Strandjord and Clayson.5 All solvents were ACS reagent grade. Solvents dissolved in corn oil prior to injection included carbon tetrachlo ride, bromotrichloromcthanc, trichloroethyl ene, methylene chloride, chloroform, tetrachlorocthylene, 1,1,2,2-tetrachlorocthanc, 1,2-dichlorocthanc, 1,1-dichlorocthanc, 1,1,1 -trichloroethanc, 1,1,2- tricholorcthanc. Freon 1 1305, Freon 1 12(fo, carbon disulfide, benzene, and toluene. The remaining sol vents were injected undiluted: ethanol, meth anol, isopropanol, /t-butanol, diethyl ether, acetone, dioxanc, //-amyl acetate, methyl iso butyl ketone, tetrahydrofuran, methyl ethyl ketone, //-butyl acetate, xylene, //-hexane, di methyl sulfoxide, A',/V-dimcthyl formamidc, and vinyl acetate. Results Prior to evaluating the hepatotoxic prop erties of organic solvents, the normal levels of serum OCT activity were measured in several species of laboratory animals. Table 1 lists the OCT activities found in animals screened in our laboratory. The mean OCT activities for the dog, cat, and rat were nearly equivalent, approximately 1 international unit (IU) of enzyme activity. The guinea pig and rabbit had slightly higher values, ap- Jannni v. I{}7'1 i a ni r l Scrum OCT Activity in Healthy Animals Species Hop Cal Rai Guinea pip Rabbil OC T Activity tr' I mean S.D.) Range 24 0.82 A 0.68 0 2.9 6 0.80 -_L 0.52 0 1.7 20 0.09 0.19 0.2 1.6 1J 1 17 1.61 0 8 9 24 2.20 t 0.9.S 0,9 4.4 '// is the number of an'inals tested. pmximately 2 IU. The range of serum OCT activity found for guinea pigs was from 0 to 8,9 IU. Values as high as 8.9 suggest that some incipient liver damage was present in a presumably healthy animal; nevertheless, the mean OCT activity was relatively low-- 2.02 rt 1.61 S.D. On the basis of results from 117 control guinea pigs, any consistent increase in serum OCT activity over 5.0 IU is assumed to indicate hepatocellular injury. Table 11 lists the solvents investigated, the doses administered, the mean scrum OCT activity 24 hours after injection, and the microscopic evaluation of liver tissue. A positive sign ( + ) for hematoxylin and eosin stained tissue sections indicates liver dam age. A negative sign ( -- ) signifies the ab sence of histopathology. For Oil Red O stained tissue sections ( + ) indicates a mod erate accumulation of lipid in liver cells. With either staining technique a () sign suggests an equivocal observation. Halogenated Methane Derivatives Carbon tetrachloride, a documented hepatotoxin, was employed as a positive control for these studies. Any solvent-induced ele vation in serum OCT activity was compared to the dose-related increases found with car bon tetrachloride. At a dose of 5 mg/kg of carbon tetrachloride the serum OCT activity was not sufficiently elevated to suggest hepa tocellular damage; however, as the dosage was increased to 25 through 150 mg/kg, the scrum OCT activity was substantially in creased. At 25 and 50 mg/kg the serum OCT activity increases dramatically, and at higher doses, the increase is not propor- SL 027253 tih'i h Sulven Cai'bm iCt riiioi. (C'l Meih\ (C'l llionii (Up I on in K1 I eliin it I 1.1.1 It I It I 11 tell it I l.l-D if , I. 2-1 It "n i.| . *( (IA A ./ .Is \V 0 1.7 i.6 o x.9 o ') 4.4 n OCT 411 0 Ul ost t hat sent in tiieless, low-- results nsistenl 5.0 1U injury. led.ihe n OCT old the r dtimMe ;toRed O modr cells. ) sign . d liepacontrol .ced elcimpared \itli car g/kg of activity si hepa dosage : Tii, the tallv ino siTum ., and at propor- Imriiciw liitliiwitil Uyywnc A\smuilinii Join mil 23 TAB IT. II I-vahialton ol the Acme Hepatoutxic Properties of Organic Solvents Solvent " Dose Mean OCT _________ Hixtopmhology //" (mg/kg) Activity Hematoxylin & Kosin1' Oil RedO` C'arhon tetrachloride (CCI,) Iltlhw lulled Melluuu Dcrivotivr.'i 4 5 3.X 4 25 37.1 X 50 63.5 4 75 53.5 4 150 64.4 (-) (+) (+) (+) (+) (-) (+) (+) (+) t+) Clilorofomi (CHCI;,) 4 150 26.4 4 300 65.7 (-) ( :) () () Methylene chloride KTCCI.,) 4 100 4 250 4 500 4 1000 4.X 1.9 2,3 4,4 (-) (-) (-) () (-) (-) (-) (-) 1J Bromoirichloromethanc ( HrCl.iC I 4 6.9 4 5 8.6 4 10 26.3 (-) (-) (+) (-) (-) <-) 1 cti'iiclllotolUIijiik''1 (CUTICHCI.,) I etraehloroethylene (CI..C = CCI.,) 1,1.1 -Trichloroethnnc (ChjCCH;) 1,1,2-T richloroethane (CICTLCHCh) Trichloroethylene (CICH = CCI,.) 1,1 -Dichloroelhane (CI.CHCH.,) 1, 2-Divliloioelhjne'' tCKTUTUI) J/iiltiL'i'iiiiit il Eihniit' IhTieaiive.'i 4 75 X 150 4 300 2.9 3.2 4.5 4 200 X.9 4 400 29.7 4 75 4 150 4 300 4 600 0.9 0.9 0.9 1.6 4 200 47.3 4 400 55.9 4 250 4 500 4 750 4 1000 4 150 4 300 4 500 4 750 3.4 3.6 4.2 2.5 1.3 1.8 1.2 3.2 4 150 3.0 4 300 3.1 4 600 34.6 (-1 <-) (-) (+) (-) (-) (-) (-) (+) (+) (-) (-) (-) (-) (- ) <-) r-) (-) <-) (-) (-) (-) (+) (-) (-) (-) (-) (-) (-) (-) (+) (-). (-) () () (-) (-) (-) (-) (-) (-) (-) 'ii is the number ol animals tested. ' ( + ) indicate hepatocellular damage. *(+) iiulic;ili:s nuKlci'iilc lipid deposition in hepaloeyles. At 500 mg/kg, all 4 animals tested died. `At 600 mg/kg, 1 of 4 annuals tested died. SL 027254 Jiwiwiv, !'>/} TAUI.li II (continued! evaluation of the Acute Hepatoloxic Properties of Organic Solvents (nig/kgi 400 800 1600 Mcttn OCT Activity 1.9 1.6 1.2 Histopalhology Hematoxylin & Hosin'1 Oil kcdO' (-) <- ) (-) (-) (:!') (+) 400 600 800 1600 2.7 3.6 3.1 2.1 (-) (-) (-) (-) ( -1 t-) ( ') (-) Aliphatic Alcohols 1000 2000 3.7 5.5 4.3 < -) 500 2.5 (-) t - ) 1000 3.2 t -) t-) 600 1200 2.6 9.8 Aliphatic Tistcrs and Ethers 1 non' 18.7 250 3.7 500 7.6 <-) (-1 () (-) (-) ( .A-) (-) (-) (+ ) 750 1500 750 1500 6.80 23.9 10.2 11.7 (-) (-) (-) (-1 0-iO (-C) (+) (-) rHistopathological data were not available. At 2000 nig/kg, I of 4 animals tested died. BAt 4000 mg/kg, all 4 animals tested died. 'At 1200 mg/kg, I of the 4 animals tested died. At 2000 mg/kg, all 4 animals tested died. JAt 500 mg/kg, 3 of 4 animals tested died. kAt 1500 mg/kg, 2 of 4 animals tested died. 'At 1500 mg/kg, 3 of 4 animals tested died. . \ nit n Solver Aeeto (Cl Mel In (Cl Metln I (Cl Hcn/c fev I nine (C, Xj let K, 1 lexa (C Oii'\ .V ii Cl N. V(ll Tctr: (f Metl (( Carl' O "/ '/ V 1 Red 6" i - )" 1 ') t+) i-) i-) (1) i-) 1-) l-i i -> ( i (. i i -1 i+) <'.) < >. i i +) (-) ,, - -...... American liultoiriiil llyytene .4ssonation Juinnnl 25 Solvent TAllI,I! II (continued) hvalualion of the Acute Hepaloloxic Properties of Orynnic Solvents Dose Mean OCT Histopathology // (mg/kg! Activity Hematoxylin & Eosinb Oil Red1 Acetone"1 <CH;lCOCII:.) Aliphatic Ketones 4 1500 4 3000 1.1 2.3 (-) (-) (+) (+ ) Methyl ethyl ketone" (CHj;COCH.,CH.,) 4 750 4 1500 4 2000 1.5 4.5 10.8 t, -) (-) (-3 ! ) (+) 1+) | Methyl isobutyl ketone" (CHyCOCH2CH (CH;,)n) 1 Benzene . fC0H,,) 4 500 4 1000 4.0 6.4 A ronuuic Hydrocarbons 4 200 4 400 4 800 4 1200 0.9 1.2 0.6 3.1 (-) (-) ( -) <-) ( -) () (-) ( =t ) (-) <-) (-) (-) Toluene (C|jHrjCTI:l t 4 150 4 300 4 600 4 1200 1.0 0.9 0.6 1.3 (-) (-) (- ) (-) (-) (-) (-) (+) Xylene" (C(iH,(CH:l)2) 4 1000 4 2000 18.4 25,2 (-) (-) (+) (+) Hexane (CII;i(CH2),CH!I) Miscellaneous Solvents 44 500 J000 13.1 21.1 Dioxanc CH.--O--CH., 4 1000 4 2000 1.1 4.1 CH .--O--CH-j N./V-Dimethyl form.imide" (HOON(CH.j12) 4 1000 4 2000 4 4000 0.9 1.6 2.2 Telrahydroftlran' (C.H,).,() 4 500 2,2 Methyl sulfoxide (C:H;,)oS() 4 750 4 1500 4 3000 1.0 2.8 0.8 Carhon disulfide' , C'S,. \ 4 100 4 200 4 400 1.0 1.3 1.3 '"At 3000 mg/kg. 2 of 4 animals tested died. "At 3000 mg/kg, I of 4 animals tested died. "At 1000 mg/kg, I of 4 animals tested died. "At 2000 mg/kg, 3 of 4 animals tested died, "At 4000 mg/kg, t of 4 animals tested died, "At 500 nig/kg, I of 4 animals tested died, sAl 400 mg/kg, 3 of 4 animals tested died. {-) (- ) <-) (-) (-) (-) (-) (-) <-> (-) <-) (-) (-) (.- ) () (+) <) (+) (-) <) (+) (+) (-) (- ) <-) (-) (+) (+) SL 027256 "hf. 26 tionatc with the dose. The elevation in scrum enzymic activity correlated well with the his tological confirmation of liver damage. Chloroform at 150 mg/kg caused a sig nificant elevation in OCT activity, and at 300 mg/kg the OCT activity doubled, sug gesting a linear dose-response relationship between scrum OCT activity and the obser vation of liver damage. The histological evaluation was. somewhat equivocal with hematoxylin and cosin stained sections at the higher dose. In regard to lipid deposi tion, equivocal results were found at both doses. Methylene chloride was studied at four dose levels from 100 to 1000 mg/kg. The OCT and the micropathological data were normal. These results arc thus consistent with the evidence that methylene chloride is not an hepatotoxin. Bromotrichloromcthanc administered at 2.5 mg/kg produced a significant increase in serum OCT activity. A dose of 5 mg/kg caused an elevation in enzymic activity that was slightly more than twice that found for an equivalent dose of carbon tetrachloride. Histological confirmation of tissue necrosis was found at 10 mg/kg with hematoxylin and eosin stained sections. Oil Red O slides were negative at all doses. On a comparative basis, this solvent appears to have slightly greater hepatotoxic properties than carbon tetrachloride. Halogenated Ethane Derivatives Eight representative halogcnatcd ethane derivatives were studied. 1,1,2,2-Tctrachlorocthanc produced no in crease in OCT activity at 300 mg/kg. At 500 mg/kg, all the animals died. Tissue sec tions at 150 mg/kg showed a moderate lipid accumulation. Liver damage was not ob served in the hematoxylin and cosin sections. These data suggest that 1,1,2.2-tctmchloroclhane hicks hepatotoxic properties even at near-lethal doses. Tetrachlorocthylcne at 200 and 400 nig/ kg produced a significant elevation in OCT Jnntnn i, / )? i activity (N.9 and 29.7, respectively). The presence of liver damage at the higher dose was confirmed by hematoxylin and eosin sections. No lipid accumulation was noted at cither dose. 1.1.1- Trichloroethanc at doses up to 600 nig/kg failed to increase serum OCT ac tivity, suggesting that this solvent has no acute hepatotoxic properties. The micropathological examination was normal. 1.1.2- Trichloroethane at 200 and 400 mg/kg produced elevations in serum OCT activity that were not dose-related. A dose of 400 mg/kg caused a 1.2-fold increase in scrum OCT activity. Apparently, doubling the dose to 400 mg/kg did not appreciably induce more liver damage. Tissue necrosis was observed at both doses, and substantial lipid deposition was found at the higher dose by staining techniques. Trichloroethylene did not . cause an in crease in scrum OCT activity even at 1000 nig/kg, thereby substantiating the fact that this solvent lacks hepatotoxic properties. Liver abnormalities were not seen at any dose; however, the Oil Red O stained tissues suggested minimal fat deposition at 750 and 1000 mg/kg. 1,1-Dichloroethane failed to elicit a sig nificant change in serum OCT activity even at the highest dose (750 mg/kg). The his tological findings were also normal. At 600 mg/kg, the injection of 1.2-dichlorocthane caused an increase in scrum OCT activity (34.6 1U). Liver damage was not confirmed by histological examination. Freon 1 12^ did not alter the scrum OCT activity even at 1600 mg/kg. Although tis sue damage was not apparent at this dose, lipid deposition was observed. The administration or Freon 113(p) at 1600 nig/kg did not elevate OCT; the ab sence of hepatocellular damage was also con firmed histologically. Ali[/Italic Alcohols When methyl alcohol was injected at a dose of 1000 mg/kg. no change in serum SL 027257 . in, OC mg res. abl } ap wh Ne llie pyl act mu i OC it I big ani int hot Ah ! sig; (l: sm bin las Ci >1 ert set of sui gir wa lip en foi (I tiv va ge 75 eh ag ,1 nwrivnn Industrial llvtticiw Ati'aniaiion Join mil OCT activity was noted; however, at 20f)0 mg/kg an insignificant increase in activity resulted. Histological data were not avail able on these animals. Neither an increase in enzymic activity nor a positive histological findings was observed when 2000 mg/kg of ethyl alcohol was in jected into guinea pigs. At 4000 mg/kg all the animals (.lied. At doses up to 1000 mg/kg with isopro pyl alcohol, no increase in serum OCT activity was noted. The histological exami nations were normal. n-Butanol productcd a slight elevation in OCT activity at 1200 mg/kg, suggesting that it possesses weak hepatotoxic properties at high doses. At 1200 mg/kg one of the four animals tested died. A slight increase in intracellular lipid deposition was noted at both dose levels. Aliphatic Esters anil Ethers Diethyl ether at 1000 mg/kg produced a significant increase in serum OCT activity (18.7 IU). Tissue necrosis was present to'a small extent, but no lipid deposition was found. At 2000 mg/kg all four animals tested died. Therefore diethyl ether can be considered to have weak hepatotoxic prop erties at high doses. For vinyl acetate at 250 mg/kg, all ob servations were normal. At 500 mg/kg three of the four animals tested died, and in the surviving animal the OCT activity was mar ginally elevated at 7.6 IU. Liver necrosis was absent, but a moderate accumulation of lipid was noted. //-Butyl acetate caused a slight increase in enzymic activity at 750 mg/kg. Two of the four animals tested died at the higher dose ( 1500 mg/kg). The mean serum OCT ac tivity of the surviving guinea pigs was ele vated at 23.9 IU. Histological results sug gested a slight accumulation of lipid. When //-amyl acetate was administered at 750 mg/kg, the serum OCT activity was elevated to 10.2 IU. Histological liver dam age and.lipid deposition were not observed. 27 At 1500 mg/kg three of the four animals tested died. The serum OCT activity of the remaining animal was elevated to about the same extent as that seen with the lower dose; although histological liver damage was absent, moderate lipid deposition was found. Aliphatic Ketones When acetone was injected at 1500 mg/ kg, no increase in serum activity was noted. At 3000 mg/kg two of the four animals tested died; nevertheless, the scrum OCT activity was still within normal limits. His tological examination revealed that no liver abnormalities were present, but moderate lipid deposition was found at both doses. Methyl ethyl ketone produced an increase in enzymic activity at 2000 mg/kg (10.8 IU). At this dose, one of four animals died. No tissue damage was seen in hematoxylin and cosin stained slides. Lipid deposition was clearly present at the two higher doses. Methyl isobutyl ketone at 500 mg/kg caused a slight but insignificant increase in scrum OCT activity. At 1000 mg/kg one animal died, and the enzymic activity was comparable to that at the lower dose. His tological liver damage and lipid deposition were not observed. A romatic Hydrocarbons Benzene or toluene administered in doses up to 1200 mg/kg produced no alterations in serum OCT activity. Liver abnormalities and lipid accumulation were not observed upon histological examination, with the ex ception of toluene at the highest dose. Xylene at 1000 mg/kg caused an increase in serum OCT activity (18.4 IU). A mod erate accumulation of lipid was noted; how ever, no tissue necrosis was found. At 2000 mg/kg, three of four animals tested tlied. The serum OCT activity in the sur viving animal was 25.2 IU. Misceliuneous Solvents Hexane when injected at 500 mg/kg caused a moderate elevation in serum OCT SL 027258 I. K.t+M.J**'.. -- .-- 28 activity (13.1 1U). At 1000 mg/kg the serum activity increased nearly twofold to 21.1 IU. Hematoxylin and cosin stained tis sues revealed no liver damage; Oil Red O staining revealed a slight accumulation of lipid at the lower dose and a moderate accumulation at the higher dose. Liver damage was absent when animals were injected with the following solvents: dioxanc (200 mg/kg), /V.^V-dimcthylformamidc (4000 mg/kg), tetrahydrofuran (500 nig/kg), dimethyl sulfoxide (3000 mg/kg), and carbon disulfide (400 mg/kg). Moder ate lipid deposition was found with N,N~ dimethylformamide, tetrahydrofuran, and carbon disulfide. The relative acute response of the solvents investigated in this study are compared in Table III. By the criteria established for this table, a low order of response is ascribed to any solvent that can cause an elevation in scrum OCT activity at a dose greater than 500 mg/kg. Moderate and high responses are ascribed to solvents that can cause an increase in serum OCT activity when the dose is between 50 and 500 mg/kg and less than 50 mg/kg, respectively. Only two sol vents caused an elevation in scrum OCT activity at low doses--bromotrichloromcthane and carbon tetrachloride; five solvents at moderate doses--chloroform, //-hexane, tetrachloroethylcne, 1.1,2-trichlorocthanc, and vinyl acetate; and nine solvents at high doses --//-amyl acetate, /i-butyl acetate, //-butyl Jtiiiuarv, 19/1 aicohol, 1,2-dichloroethane, diethyl ether, methyl alcohol, methyl ethyl ketone, methyl isobutyl ketone, and xylene. With many sol vents in this last group a dose of 1000 mg/kg or greater was necessary to produce an ele vation in serum OCT activity. The remain ing solvents did not cause any perceptible liver damage as determined by the OCT assay or by histological examination. Discussion It is well documented in the scientific literature that the measurement of scrum OCT activity can be employed as a sensitive and specific liver enzyme test. The results obtained here have demonstrated with the use of a known hepatotoxin. carbon tetra chloride, that, as the solvent dosage was in creased, hepatic tissue was damaged, and this resulted in a concomitant increase in scrum OCT activity. For many solvents good agree ment was found between the increase in scrum OCT activity and the histological verification of liver damage. With sonic organic solvents, it seemed that the initial insult to the liver was sufficient to block the occurrence of additional liver damage by higher doses of solvent. For example, when carbon tetrachloride was administered at a dose above 50 mg/kg, no increase in serum OCT activity was observed. It has also been shown with some solvents that an increase in serum OCT activity often precedes the histological verification of liver TABLK lit Relative Order of Hepalotoxicity" Low Moderate High Amyl acetate Butyl acetate Butyl alcohol 1,2-Dichlorocthnnc Diethyl ether Methyl alcohol Methyl ethyl ketotic Methyl isohulyl ketone Xylene Chloroform Hexane Tctrachlorocthylcne I, 1, 2-Trichlorocthanc Vinyl acetate Bromoirichloromethane Carbon tetrachloride aLo\v indicates an elevated OCT at a dose greater than 500 mg/kg. Moderate indicates an elevated OCT at a dose between 50 and 500 nig/kg. High indicates an elevated OCT aL a dose less than 50 mg/kg. SL 027259 .I d. n d. di w ll H Ci P h j* V p ( r American Industrial Hygiene Association Journal damage. This demonstrates that the sensi tivity of the OCT assay nitty be adequate to detect incipient liver injury; a particularly desirable feature when evaluating low level exposures to hepatotoxins. In most cases, when the serum OCT activity was greater than 5 IU, there was a good agreement be tween the enzymic assay and the histological confirmation of liver damage. An increase in lipid deposition in liver parenchyma cells was often observed with high doses of several solvents. In the ma jority of cases no additional liver pathology was apparent in these tissue sections. In conclusion, it has been demonstrated that the OCT assay has high specificity and senistivity for detecting hepatocellular injury. Of the thirty-three solvents evaluated, two produced an elevation in serum OCT activity at relatively low doses, five at moderate 29 doses, and nine at relatively high doses. Acknowledgment We wish to thank Mr. Daniel S. Thomas for his technical assistance throughout this study. References 1. Harman, T. E.: Enzyme Handbook, Vol. I, p. 280, Springer-Verhig New York (1969). 2. Reichard, P.: Ornithine Carbamyl Transferase from Rat Liver. Acta Chein. Scand. ii:51l (1957) 3. Reichard, P.: Ornithine Carbamyl Transferase Activity in Human Tissue Homogenates. J. Lab. Clin. Med. 56:218 (I960). 4. Reichard, H., and P. Reichard: Determinations of Ornithine Carbamyl Transferase in Serum. J. Lab. CUn. Mad. 52:709 (1958), 5. Strandjord, P. E., and K. J. Clayson: An Auto matic Method for the Determination of Orni thine Carbamyl Transferase Activity. J. Lab. Clin. Med. 67:1.54 (1966). ReceiveJ June 13, 1973 Recent Actions on ANSI Standards The billowing standards have been acted upon recently by the American National Standards Institute and will be of interest to'industrial hygienists: S3.6-1969 Specifications for Audiometers. This standard was reaf firmed, November 15, 1973. 737.12-1973 Acceptable Concentrations of Toluene. This standard is a revision of the 1967 standard. 7.37.I6-1967 Acceptable Concentrations of Formaldehyde. This standard was reaffirmed, November 12, 1973. 7.37.3S-I973 Acceptable Concentrations of (Fluorocarbon 12) Dicliloroilijlitoronie/hane. This new standard was approved November 12, 1973. N 13.8-1973 Radiation Protection in Uranium Mines. This new stand ard has been published recently, Bibliography of Metric Standards Companies considering the use of the metric system can obtain guidance on the availability of metric standards from a recent publication of the American National Standards Institute. Single copies of A Bibliography of Metric Standards, ANSI Special Publication II, may be obtained without charge from the American National Standards Institute, 1430 Broadway, New York, New York 100IS. Sh 027260 Enclosure 1 NIOSH Recommended Precautionary Monitoring and Control Procedures for Polymerization Processes Involving Vinyl Chloride I. General Housekeeping Procedures A. The spillage of Vinyl Chloride and Polyvinyl Chloride in and around the production facilities should be controlled as follows: 1. Housekeeping procedures should be implemented to assure immediate removal of VC and PVC material around polymerization operations including drying, packaging and loading operations. 2. Recovered PVC material which is to be packaged should be stored in closed containers. 3. Waste PVC material should be stored in closed containers and consideration should be given to the adequacy of its disposal and/or destruction. Care should be taken in the storage of closed containers to insure that unsafe conditions do not result from an internal build-up of pressure in the container. B. Inventories of beginning and recovered quantities of VC and quantities of PVC produced, packaged and recovered should be made to determine losses and probable areas affected. C. PVC material should be removed from overhead structures and conduits where it tends to collect. D. Consumption of food should be permitted only in separate facilities provided for this purpose, and no food products should be permitted elsewhere in the polymerization facility. II. Protective Clothing A. A daily change of protective clothing including full coveralls, or the equilvalent, should be provided each employee in areas where possible exposure to VC PVC could occur. Clothing contaminated hy accidental spills should be changed as soon as feasible. B. Protective gloves and footwear, or footcovers, should be worn as appropriate in those PVC operations where exposure to PVC material is possible. C. Protective head covers should be worn during PVC operations as appropriate (e.g. hard hats in those areas where physical protection of the head is necessary and hair coverings, or the equilvalent, in SL 027261 operations such as cleaning of polymerization reactor tanks and packaging operations). D. Where emoloyees are engaged in maintenance or cleaning operations of polymerization reacrors tames they should wear full impervious suits to guard against skin contact of PVC material and VC vapors. Procedures for safe entry to confined spaces should be observed. III. Showers Showers at the termination of an eight-hour work shift should be mandatory for all workers with possible contact with VC-PVGt IV. Monitoring A. Environmental Monitoring 1. In-plant environmental monitoring programs should be implemented; and where workers are required to enter polymerization tanks, a survey of the VC concentrations should be made in the reactors immediately after opening, immediately prior to entry of maintenance personnel and during the tank cleaning operation. 2. Data obtained from the environmental monitoring program should be used to indicate those areas where efforts should be directed to reduce airborne levels of 'VC. 3. Positive programs to initially control VC levels well below the present Federal Standard of 500 ppm should be developed Csorae companies are targeting at 50 ppm) and efforts to further reduce levels should be given consideration concomitantly with the necessary modifications in engineering and design controls. 4. The concentration of VC in exhaust ventilation should be determined to estimate the amount of VC lost during operations and the possible exposure of personnel immediately outside the facility. 5. Monitoring for PVC particulate material should be accomplished to estimate the degree of exposure to this substance throughout the facility. B. Personal Monitoring 1. Integrated, eight-hour personal monitoring samples should be obtained from those employees considered to receive the greatest exposure to YC or PVC. SL 027262 2. Breathing zone samples should be obtained to complement environmental monitoring program for VC, and similar samples should be obtained for PVC. Respiratory Protection Because VC is a gas under ambient conditions and PVC is a solid under these conditions, it is recommended that respiratory protection for employees take these circumstances into consideration. A. Where employees are engaged in cleaning and maintenance operations inside polymerization reactors they should be equipped with an atmosphere-supplied respirator in order to protect against both VC vapor and pVG parfcieulates. B. During housekeeping procedures and packaging operations where the possibility of PVC dust inhalation is a factor, it is recommended that an air-purifying respirator equipped with a mechanical filter designed to remove particulate material be worn. Respiratory protective devices which meet this requirement, as well as protect against VC vapors in concentrations less than 0.1% have been approved by the NIOSH Testing and Certification Laboratory and bear the numbers: TC-23C-4Q, TC-23C-47, TC-23C-48. C. Where employees are engaged in transfer operations of VC from railway tankcars to storage facilities or at any similar transfer point which requires manual operations, they should wear a selfcontained breathing apparatus SCBA) during such operations to guard against unexpected release of VC during such operations. D. Other operations involving possible exposure to VC or PVC should be evaluated as the individual situation exists and respiratory protection provided as appropriate. SL 027263