Document Exb0JgbOOM0vn4vaQeYRNRL7x

VTJYLIDEhE C-JLCRIDE and vewlidbie o2xmi)e-'/biyl chlcrtde ccpcomers VinvLider.e chloride 1. Chemical and Physical Data 1.1 Synonyms and trade names j Chem. Abstr. Services Reg. No.: 75935-4 I Chem. Abstr. Name: 1,1-Dichloroethene 1,1-Dichloroethylane; ggyw-dishloroethylene Sconatex 1.2 Structural and molecular formulae and, weight Mol. wt: 97.0 1.3 Chemical and physical properties of the pure substance Fran Weast (1976), unless otherwise specified (a) Description: Clear liquid with a sweet odour (Hardie, 1964; Windholr, 1976) (b) . Soilina-point: 37C * (c) Melting-point: -122.1C (d) Density: d^ 1.218; vapour density (Anon., 1972) (e) Refractive index: 20 1.4249 3.4 (air = 1) (f) Spectroscopy data: X vapour <200 nm; infra-red, nuclear magnetic resonance, and mass spectral data have been tabulated (Grasselll & Ritchey, 1975) (g) Solubility: Insoluble in water (0.04% wt/vol. at 20C); miscible with most organic solvents (Hardie, 1964) XL 5uh"J~\+ ~ ?r ' ':-'ar in Ross_ v ` , " ',A' - 4337 14-h .3:1 :- " Calcasieu Parish, Louisiana CMA 01561? (h) volatility: Vapour pressure is 400 mn at 14.3C (Perry & Guitar., 1973) (i) St-ahllicy: Flash-point (closed cup], -17C (Anon., 1972); easily polymerized at temperatures above QC in the presence of oxygen or other catalysts (JVindholz, 1976) (j_) Conversion factor: 1 pan in air is approximately 4 ng/m3 1.4 Technical products and impurities Vinylidene chloride is ccrtmercially available in the US with the foliating typical specifications: vinylidene chloride, 99.6 wt %; acetylene, 25 mg/kg max.; other chlorinated hydrocarbons, none exceeding 0.25 wt %; acidity (as hydrogen diloride), 10 irgAg max.; peroxide (as hydrogen peroxide), 10 rng/kg max.; water, 50 mg/kg max.; and inhibitor (mcnorethyl ether of hydroquincre), 180 to 220 mg/kg (PPG Industries, 1975). Typical specifications fear vinylidene chloride produced in Japan are as follows: specific gravity (d^), 1.2129; melting-point, -122.1C; boiling-point, 33.4C; and refractive index (n^), 1.4249. 2. Production, Use, Occurrence and Analysis 2.1 Production and use (a) Production Vinylidene chloride was first prepared by Begnault in 1838 by the reaction of trichloroethane and alcoholic potassiirn hydrojdde (Beinhardt, 1943). Although vinylidene chloride may be prepared by several methods, it is catuercial ly produced in the 05 and Japan by the dehydrochlorination (using sodium hydroxide or liim) of 1,1,2-trichloroethane, derived frcm ethylene dichloride. The resulting crude vinyl idene chloride is purified by washing, drying, and fractional distillation. Inhibitors are normally added at this point (Wessling & Edwards, 1971). r----- -- -- --7 T iTi ' Subject^ - . _ ' Boss v. J_ ` i4th ; - - ` .. . Calcasieu Parisu, 'r in ' -37 ana CMA 015618 The oarmercial production of vinylidene chloride was dependent on the developnent and conreroialization of vinylidene chloride copolymers, and was first reported in the US in 1940 (US Tariff Conrission, 1941). It is estimated that tr*o US conpanies produced a cottoned total of 70 million kg vinylidene chloride in 1976 aoi that cne of these conpanies nanufactursd an additional 50 million kg for captive use as an unisolated intermediate in the production of 1,1,1-trichloroethane. US airports ard exports of vinylidene chloride are negligible. Vinylidene chloride has been produced carrrertialiy in Japan since 1951, In 1976, three conpanies produced a carbined total of 28.1 million kg. Japanese imports and exports of vinylidene chloride are negligible. (b) Use Excluding the amount used as an unisolated intermediate in the production of 1,1,1-trichloroethane, more than 90% of the vinylidene chloride produced in the US and Japan is used in the production of copolymers of high vinylidene chloride ocntent, the ether major monomer usually being vinyl chloride. For a discussion on the uses of vinylidene chloride-vinyl chloride copolymers and other vinylidene chloride-based polymers, see p. The remaining 10% or less of the vinylidene chloride produced is used in the manufacture of irrdacrylic fibres, which are largely based on acrylonitrile with snail amounts of vinylidene chloride and other monansrs. For a discussion on the uses of these fibres, see "Acrylic and modacrylic fibres", p. The American Conference of Governmental Industrial Hygienists (ACGIH) reaocnends that an eiployee's exposure to vinylidene chloride does not exceed an eight-hour time weighted average of 40 mg/in3 (10 peto) in the workplace air in any eight-hour work shift of a forty-hour writ vreek. During any fifteen minute period, the ACGIH proposes an absolute ceiling concentration limit of 80 mg/m3 (20 pfm) provided the daily threshold limit value (in forrwt of eight-hour time weighted values) is not exceeded (ACGIH, 1976) CONFIDENT I A.L Subject Or''4-!!1 in Boss v. C 14th J-:-: Calcasieu Farish, LLouisiana CMA 015619 2.2 Occurrence Vinylidene chloride is nor known to occur as a natural product, (a) Air in polymerization processes (e.g., polyvinyl chloride) have been reported to be exposed to vinylidene chloride concentrations (Jaeger, 1976; Ott si at., 1975) in amounts of less than 20 mg/fti3 (5 ppm) and most frequently to trace amounts (Kramer & Mutchler, 1972). Vinylidene chloride at levels of 3 mg/m3 (2 ppm) has also been reported to be a contaminant of submarine and at levels of 0-2 ppm of spacecraft atmospheres (Altman & Dittmer, 1966). Emissions of vinylidene chloride in the US in 1974 have been estimated at 1.52 million kg from monomer synthesis operations (reduced to 277 thousand kg by new control technology in late 1975), 308 thousand kg fron polymer synthesis operations, and 13.8 thousand kg fran polymer fabrication opera tions (Hushon & Kornreioh, 1976). (b) Water Vinylidene chloride has been detected in effluent discharged fran chemical manufacturing plants in The Netherlands at a concentration of 32 ug/1 (Eurocop-Cost, 1976) and in effluent discharged by chenical and latex manufacturing plants in the US. It has also been identified in well, river, and raw water in the US (Shackelford & Keith, 1976). file highest reported concentration of vinylidene chloride in the US finished drinking water was 0.1 yg/1 (US Oivironmental Protection Agency, 1975). (c) Other Vinylidene chloride has been found as an impurity in trichloroethylene mananer (limit of detection, 5 mg/kg) (Kiezel st al., 1975; sassu st at., 1968), and at a level of 0.011% in eecirercial chlorcprene (Kurginyan & Shirinyan, 1969). Ccmnercial household and industrial saran films have been analyzed for residual vinylidene chloride monomer. Six rolls of household film had in ' '337 ' Tt ana CMA 015620 monomer concentrations ranging from 6.5 to 10.4 mgAg, with an average of 8.3 mgAg* There vere no significant differences in sanplas taken from the beginning (outside) or the end (inside) of each roll. The industrial film shewed levels ranging from 10.3 to 26.2 mgAg with levels increasing fran. the beginning to the end of the roll CBirkel at at., 1977). 2.3 Analysis Vinylidane chloride has been identified in air by trapping in pyridine ard colorimetric determination of the cyanine obtained by reaction with barbituric acid cr aniline. The limit of detection was 10 mg/tn3 (2-5 ppm) (Grqnsberg, 1975). Gas chromatography has been used to determine vinylidene chloride as an ircpurity (1) in trichloroethylene (Vlasov & Bodyagin, 1970) and (2) in vinyl chloride mcnaner (Sassu at at., 1968) with a limit of detection of 5 mgAg (Kiezel at at., 1975). The same thod has been applied to the detection of free vinylidene chloride in latex (Bollini at at., 1974). Sampling techniques using activated carbon as adsorbed with subsequent solvent or thermal desorption and gas chromatographic analysis have been evaluated to determine vinylidene chloride in industrial atmospheres (Severs & Skary, 1975). A satrpling technique for concentration of air pollutants including vinylidene chloride on different gas chromatography has been evaluated. The ccnpounds were thermally desorbed and analyzed by gas chrcnatography using flams ionization detection; the limit of detection is 4 ug/m3 (1 ppb) (Russell, 1975). Vinylidene chloride has been detected in saran films by gas chromato graphy with electron capture detection and mss spectrometry confirmation. The limit of detection of the method is 5 mgAg (Birkel at at., 1977). CONFIDENTIAL Subject cc 7rotec: Ross v 14 V": ' Calcasieu Parian, CMA 015621 in i D istrict* Court P arish, Louisiana ef* a =) to 3. 3lological Data rtelevant to the Evaluation of Carcinogenic Sisk to Hunans 3.1 Carcinogenicity and related sf-sdi'es in animals1 (a) Oral administration Bar: In a preliminary report of an ongoing study, groups of 50 rale and 50 female Sprague-Dawlsy rats were adninistared 5, 10 or 20 rg/kg bw vinylidene chloride in olive oil by stonach tube once daily, 4-5 davs/week, far 52 weeks; 1 carcinana of the Zymbal gland was observed in a rat treated with the 10 mg/kg fcw dose, A ccntrol group of 100 male and 100 female rats were given olive oil alone. At the time of reporting, the rats had been observed for 93 weeks after the start of treatment (Maltcni et al., 1977). (b) Inhalation exposure Mouse: in an experinent still in progress at the time of reporting, groups of Swiss mice, 9 or IS weeks of age, were exposed 4 hours/day, 4-5 days/week, to vinylidene chloride vapours in air at concentrations of 800, 400, .200, 100, or 40 mg/m3 (200, 100, 50, 25 or 10 ppm). Due to toxicity, mice treated with 200 (60 tales and 60 fatales) or 100 ppm (30 males and 30 females) were e^osed for only 2 days and those treated with 50 pen (30 tales and 30 females) for cne week cnly . At the 25 pan level, 30 males and 30 fdales were initially exposed and a furtter group of 120 male and 120 faiale mioe was started at the same exposure level and treatment of both groups continued for 52 weeks; at the tiTM of reporting, 98 weeks, 24/150 tales and 1/150 females had developed adenocarcinoias of the kidney, often bilateral. Che male had a kidney aderocardraoa in the group treated with 50 ppm far one week. Mo such tumours had occurred in mice exposed to 10 jxm for 52 weeks or in 330 controls (both groups observed until 98 wreeks of age) (Maltcni, 1977; Maltoni et al., 1977). -r <(, VC- 'Aj- 1The Working Group was aware of ongoing studies to assess the carcino genicity of vinylidene chloride in mice by skin application1 and in mice and rats by oral adninistration (IARC, 1978). The Working Group was also aware of ongoing oral (drinking-water) and inhalation studies using rats in which the data analysis was not acnplete (Barrgy at al., 1977). Further inhalation studies on two strains of rats wrere also incarplete (Viola & Caputo, 1977). CMA 015622 A group of 36 male and 36 female CD-I mice, 2 monchs of age, were exposed to 220 ag/a3 (35 ppa) vinylidene chloride in air 6 hours/day, 5 days a week for 12 months, at which time the experiment was terminated. Two aales died early in the experiment and were replaced by healthy mice; 2 males were killed in the 9th month of treatment and one female during the 10th month of treatment. 3ronchiolo-alveolar adenomas occurred in 6 mice and angiosarcomas of the liver occurred in 3 mice created with vinylidene chloride; no such tumours occurred in controls. Three hepatomas and 2 skin keratoacanchomas were also reported to occur in treated mice (Lee st at., 1977, 1973) [The Working Group noted the short duration of the experiment]. Rat: A group of 36 mala and 36 female CD rats were exposed to vinylidene chloride in air at a concentration of 220 mg/m3 (55 ppm) 6 hours/day, 5 days a week for up to 12 months, at which time the experiment was terminiated and all survivors killed. Two rats developed angiosarcomas, one in the mesenteric lymph node and one in the subcutaneous tissue. Such tumours did not occur in controls (Lee st at., 1977, 1973) [The Working Group noted the short duration of the experiment]. One group of 60 male and 60 female Sprague-Dawley rats were exposed to 800 reduced to 600 mg/m3 (200 reduced Co L50 ppm), and 4 groups of 30 female and 30 male Sprague-Dawley rats were exposed to 40, 100, 200 or 400 mg/m3 (10, 25, 50 or 100 ppm) vinylidene chloride in air for 4 hours/day, 4-5 days a week for 52 weeks and observed for up to 82 weeks (time of reporting). The animals were 16- weeks old at the start of treatment. An Increased incidence of mammary fibroadenomas and carcinomas (40-60Z) was reported when compared with 100 male and 100 femala controls (32Z). No dose-response relation was found. In addition, one Zymbel gland carcinoma was seen in one rat treated with the 100 ppm dose (Maleoni at at., 1977). Hamster; In a study still in progress, no tumours had occurred at 74 weeks among a group of 30 male and 30 female Chinese hamsters, 28 weeks of age, exposed to 25 ppm vinylidene chloride in air 4 hours/day, 4-5 days a week for 52 weeks (Maltoni at at., 1977). ~ .~i -j r t ^ )T T TAT .4 "-1 Subject Boss 7 ` _ 14th Jui: Calcasieu . ~i3t Parish,, -- - 1n ... . ^- 4337 Court Louisiana CMfi 015623 3.2 Other relevant biological data The adverse biological effects of vinyiidene chloride have been reviewed (ERA, 1976; Haley, 1975; Warren 4 Ricci, 1978). (a) Experimental systems Toxic effects Results reported for acute toxicity studies on vinyiidene chloride have been highly variable, the lethal concentrations are dependent on the dietary parameters (fed or fasted animals) and on the hepatic glutathione content which exhibited significant variations in diurnal rhythm (Jaeger et al. , 1972a, 1974). The LCjo by inhalation for a 4 hour exposure was 40-60 mg/1 (10,000-15,000 ppm) in fed rats and 2-10 mg/1 (500-2500 ppm) in fasted rats (Jaeger et al., 1973b); the minimum lethal concentration was 10,000 ppm for a 24 hour exposure for fed rats (Jaeger et al. , 1974), The LCjo of vinyiidene chloride for rats following exposure for 4 hours and observation for 2 weeks was 25.4 mg/1 (6350 ppm) (Siegel et al., 1971). The oral LD50 in mice was 200 mg/kg bv (Jones 4 Hathvay, 1978a). The oral LDsa was 1500 mg/kg bw in normal rats and 30 mg/kg bw in adrenalectomized rats (Jenkins et al., 1972). In dog, the oral minimal lethal dose was 5750 mg/kg bv and the i.v. minimal lethal dose was 225 mg/kg bw. The s.c. minimal lethal dose in rabbits was 3900 mg/kg bw (Barsoua 4 Saad, 1934). Death was due to vascular collapse and shock (Jaeger et al., 1973b). Inhalation studies using rats, guinea-pigs, dogs, rabbits and monkeys with exposures at mean level of 189 mg/m1 (48 ppm) for 90 days produced significant mortality and liver damage but no changes in haematologlcal parameters (Prendergast art at., 1967). Inhalation of vinyiidene chloride for 20 6-hour exposures at 2 mg/1 (500 ppm) caused nose irritation, reduced weight gain and hepatic histopsthological changes in rats (Gage, 1970). In fasted rats exposed to 0.8 mg/1 (200 ppm) for 4 hours, liver parenchymal cell injury was observed (Reynolds et at., 1975). Minimal liver changes characterized by an increase in cytoplasmic vacuolization of occasional individual hepatocytes were noted in Spragueawley rats treated with 6-8 mg/kg bv/day for 90 days vinyiidene chloride CONFIDENTIAL Ro s j v 14 th Calcasieu Pari - \3Z? - ' urt L viloiana CMA 015624 given at a concentration of 200 mg/l in their drinking-water. In a status report an a two-year study with rale and female Sprague-Dawley rats receiving 16-40 mgAg bw/day (200-230 mg/l), 3-20 mg/kg bw/day (100-120 mg/l) and 5-12 mgAg hw/day (60-70 mgA) in their drinking-water, no toxicological effects were noted except for a non-dose related decrease in survival of male rats only at IB and' 24 months. ?ats exposed for 30 or 90 days to doses of 0.1-0.3 mg/l (25-75 pern) in air showed minimal toxicologi cal effects in the liver (Norris, 1977). Vinylidene chloride affects the activity of several liver enoyraes, notably it decreases hepatic glucose-6-phosphatase and increases serum alanine a-ketoglutarate transaminase (SAKT) . It increases the liver con tent of triglycerides and decreases that of glutathione. Decreased hepatic glutathione concentrations increase the lethality and hepatotaxicity of vinylidene chloride (Jaeger et zi., 1973a,c). The microsanal enzyme inducers, phenobarbital and 3-methylcholanthrene, increased the lethality of vinylidene chloride inhalation (Carlson & Puller, 1972). Vinyl chloride, when administered simultaneously with vinylidene chloride, prevented the injury associated with vinylidene chloride inha lation in fasted rats (Jaeger, 1975). Various epoxidps (1,1,1-trichloropropane-2,3-oxide; 2,3-epcxyprcpanl-ol; styrene oxide; butadiene monoxide; and cyclohexane oxide) enhanced the hepatoxicity of vinylidene chloride in male rats and decreased the acute oral ID (Andersen & Jenkins, 1977). S3 Sftfaryotoxicity and teratogenicity Pats were given vinylidene chloride either as 200 mg/l in the drinkingwater or as 20-160 ppm (80-640 mj/m3) 7 hours/day by inhalation an cays 6-15 of gestation. Rabbits were given the same dose by inhalation on days 6-18 of gestation. No teratogenic effect was seen both in rats or rabbits. Sane evidence of arbryutaxicity or foetotoxicity was observed in both rats and rabbits exposed to vinylidene chloride by inhalation; these effects were associated with maternally toxic levels of exposure (Norris, 1977). COKFIygNTIAL Subject to Protective Order la Boss v. Cono-o. Ir.G.. !To. 90-4837 CMA 015625 C O N F ID E N T IA L C:oO 4-> s- 1 3 I - d > Kp) wn o M, Absorption, distribution, excretion and aecabolism la rats, as the dose level of radioactive viaylidene chloride is increased from 1-50 ag/kg bv oral, or from 0.04-0.3 mg/1 (10-200 ppm) inhalacioa, the metabolic pathway becomes saturated so that percentage wise, less of the dose administered is metabolized and more is eliminated fit the lungs as viaylidene chloride. At the 1 mg/kg bv oral dose and the 10 ppm inhalation dose, there was no difference in elimination by fed versus fasted rats. At 50 mg/kg bv oral or 200 ppm inhalation, there was a significant increase in the excretion of vinylidene chloride via the lungs and decrease in urinary excretion of radioactivity in fed versus fasted racs (Norris, 1977). The main eliminative route for 1"c-vinylidene chloride after intragastric, i.v. or i.p. administration to rats is pul monary; both unchanged vinylidene chloride and related CO2 are excreted by that route and other vinylidene chloride metabolites via the kidneys. Biocransformation of viaylidene chloride gives chiodihydroxyacetic acid and an .V-acetyl-5-cysteiaylacecyl derivative as major urinary metabolites together with substantial amounts of chloroacetlc acid, dlthiohydroxyacetlc acid and thiohydroxyacetic acid (Jonas & Hathway, 1978b). Mice metabolize a greater proportion of an oral dose of 50 mg/kg bv vinylidene chloride than rats. Mice (but not racs) excrete a small amount of <V-aeecyl-5-(2-carboxymethyl)cysteine and they excrete more .V-acetyl-Scysteinylacetyl derivative chan do rats (Jones & Hathway, 1978a). Metabolic conversion of vinylidene chloride into an epoxide which can rearrange to the correspondent acyl chloride has been proposed (Henschler, 1978; `Jones & Hathway, 1978a,b). Mutagenicity and other short-term tests Vinylidene chloride at concentrations.of 22 and 202 (20,000-200,000 ppm) in air produced reverse mutations in Salmonella typhimurium TA100 and TA 1530 in the presence of 9000 g supernatant from liver, lung and kidneys of mice and rats (3artsch et al., 1975) and in one human liver specimen (3artsch et al., 1976). At a concentration of 52 (30000 ppm) in air, it is mutagenic in 5. typkimujnian TA1535 in tha presence of a S-9 mix of liver or kidney from mice and rats pretreated with Aroclor 1254. It was weakly mutagenic in the presence of a S-9 mix of liver from a human subject who had been receiving long-term phenobarbital medication (Jones & Hathway, 1978c). Reverse mutations were induced in E3eheriehia coli K 12 by vinylidene chloride solution in the presence of liver microsomes from urine precreated with phenobarbital (Greim et al., 1975). CMA 015626 Vinylidene chloride was not mutagenic in the dominant lethal test in tale Ol mice exposed by inhalation to 40, 120 and 200 mg/m3 (10, 30 and 50 pan) for 6 hours/day for 5 days (Anderson st cl., 1978). (b) Humans Acute exposure to high concentrations of vinylidene chloride in air results in central nervous systsn^ depression and narcosis. Repeated exposures to lew concentrations is associated with liver ar.c renal dysfunction. Skin contact with vinylidene chloride causes irritation which may be partly due to the hydroquinene mcnanethyl ether inhibitor. Contact with the eye causes conjunctivitis and transient corneal injury (Irish, 1963). 3.3 Case remits and eoidaniological studies Ott st cl. (1976) investigated the canoer risk among a cohort of L38 writers exposed to vinylidene chloride fchere vinyl chloride was not used as a copolymer. The authors reported that there were ro findings statistically related or individually attributed to vinylidene chloride exposure in this cohort [The working Group noted that because 27 workers were lost to follow-up but considered alive in the analyses, and that because 55 people had less than 15 years since first exposure and only 5 deaths were observed, these factors preclude any judgement on the findings]. Vinylidene chloride-vinyl chloride copolvers 1. qremical and Physical Gate 1.1 ^vnenyms and rirede names Chem. Abstr. Services Peg. Ho.: 9011-06-7 Cham, Abstr. Name: 1,l-Didiloroethene polymer with chloroethane Chlorcethylene-1,1-dichloroethylene polymer; 1,1-dichloroethylenenrrx^johloroethylene polymer; 1,1-dichloroethylene polymer with chlorcethylere; vinyl chloride copolymer with vinylidene chloride; vinyl chloride-1,1-dichloroethylene copolymer; vinyl chloridevinyl idpne chloride copolymer; vinyl chloride-vinyl idene chloride polymer; vinylidene chloride-vinyl chloride polymer ^ -|r Hess 7 14th i Calcasieu Parish, in 337 ' *rt Louisiana Ol5 627 Breon 202; 3raou CS 100/30; Daran; Daran CR 6795H; Dow 874; Dow Latex 874; ET 67; Geon 222; Geon 652; IXhS 1; KhS 596; Kurehaloa AO; Laplen; Latex 5VXh; Polyco 2611; QX 2163; Saraa 633; Saran 746; Saran Resin 683; S? 489 SVXh 1; SVXh 40; U? 925; Velon; VIKh 65; Vinidea 60; VKhVD 40; Viaidan 60 1.2 Structural and molecular formulae and molecular weight in combination with ^Wx- (C2H3C1)y Mol. wt: 10,000 to 100,000 1.3 Chemical and physical properties of the copolymer (a) Description: Crystalline or amorphous powder depending upon the content of vinylidene chloride (Anon., 1973) (b) Melting-point: 183-195C (Wessling & Edwards, 1971) (c) Solubility: Soluble in tetrahydrofuran, 1,4-dioxane, cyclohexanone, cyclopentanone, chlorobenzene, and dichloro benzene (Wessling & Edwards, 1971). (d) Stability: Resistant to sunlight and weathering; y-rays cause cross linking and chain scission (Wessling & Edwards, 1971) 1.4 Technical products and impurities Vinylidene chloride-vinyl chloride copolymers are commercially available in the US in several forms: resins, latexes, films, and fibres. The ideneley and amount of impurities in these products are noe available. In general, products obtained by emulsion polymerization (i.e., latexes) have varying amounts of additives used in the polymerization process, such as initiators, activators, and surface-active agents. CMA 015628 The vinylidene `dlloride-vinyl chloride copolymers ocorercially available in Japan are believed to be based on 80-90% vinylidene chloride units. 2. ?roduction. Use, Ccourrer.ee and Analysis 2.1 Production and 'use (a) Production The polymerization of vinylidene chloride was first observed by Begnault in 1838. During the 1930's in the US, the cairoerical development of the polymer began. The pure honopolymer is difficult to fabricate because its softening point is very close to its decarpcsitian point. The discovery that inclusion of small amounts of other monaners lowered the softening point led to the commercial introduction in 1940 in the US of the family of vinylidene chloride copolymers now known as saran (Gabfcett & Smith, 1964). The copolymers presently of earnercial interest in the US are: vinylidene chloride-vinyl chloride (Saran S), vinylidene chlorideaUcyl acrylate (Saran C), and vinylidene chloride-acrylonitrile (Saran F). Because saran has now cane to be a generic term in the US for copolymers of high vinylidene chloride content, and because these copolymers are ai often referred to as simply 'polyvinylidene chloride', the aanpositicn of the copolymers described in the literature is not always known. Vinylidene chloride-vinyl chloride copolymers are presently produced in the US by free radical processes emulsion or suspension. In Japan, they are* produced by an emulsion process. The amilsian process produces a polymer latex which can be used directly (usually with additional stabilizing ingredients) or the polymer can be recovered, usually by coagulation with an electrolyte, foHewed by washing ard drying. The emulsion process has the advantage of producing a higher molecular weight polymer than the suspension process and the disadvantage of the relatively high concentration of additives which may affect sore of the properties of the polymer. Suspension polymerization is usually used for oopolymars used as moulding and extruding resins. M? "37 Loui-3 j.a.n& CMA 015629 Four US manufacturers now produce vinylidene chloride-vinyl chloride copolymer resins, latexes, and films, and one manufacturer produces vinylicene chloride-vinyl chloride fibre. US production of virylidere chloride ccpolyrers in 1977 has been estimated at 68 million kg (Anon., 1977). Vinylidene chloride-vinyl chloride copolymers were first produced crxreroiaily in Japan in 1951. In 1976, four Japanese carparJ.es produced an estimated 31.9 million kg vinylidene chlcride-vinyl chloride copolymers, 2.7 million kg of which was fibre and the remainder, Latex and film. (b) Use vinylidene chloride-vinyl chloride copolymers have gained wide use due to their barrier properties to water and gases, their resistance to oil, grease, chemicals, and sunlight, their flexibility, and their heat sealability. In addition, the high chlorine content imparts fire retardancy to the product. Vinylidene chloride-vinyl chloride copolymers are used in the form of films for focd packaging (the largest use); coatings for cellophane, paper and other surfaces; fibres; and tubes and pipes. Applications for vinylidene chloride-vinyl chloride copolymers in food packaging include; household food wrap; industrial food wrap for dnro liners, cheese, luncheon meat, and sausage; shrink, film for beef, poultry, and cheese; and in Laminations for cap liners, cosmetics, and luncdiecn meat. A relatively new use is in a coextruded multi-layered film of polyethylene on a vinylidene chloride-vinyl chloride copolymer core <Both, 1976). Vinylidene chloride-vinyl chloride copolymers are used as coatings in many applications, and as such are used in two farms: solvent-soluble resins, and water dispersions or latexes. The solvent-soluble resins are used to coat other polymer films such as cellophane (the largest single application), paper drinking cups and plates, and paperboard cartons. Other uses include interior coatings for ship tanks, railroad tank cars, and fuel storage tanks, coatings for steel piles and structures, binders in coatings for magnetic tapes, audio tapes, video tapes, and ccnputer tapes (Both, 1976; Wessling & Edwards, 1971) CONFIDENTIAL Subject to Boss v. _C; ' 14th : Calc'f' ?"*' o-v - .. j. i:ia CMA 015630 The Latexes are used for seating paper for use in packaging potato chips, pretzels, cereal, and cake mixes, for coating polypropylene and ether plastics for packaging and for single serving containers; and for coating papertcard for packaging candy, baked goods, and frozen and refri gerated items. Other applications include use as binders for paints and roTwoven fabrics and as an additive to cement to rake high-strength mortars and concretes (Roth, 1976; Viessling & Edwards, 1971). Extruded fibres cade fran vinylidene chloride-vinyl chloride copolymer resins are used in a wide variety of applications where resistance to sunlight and chemicals is required (e.g., autanotive seat covers, outdoor furniture, agricultural shade cloth, and filter fabrics) (Vfessling & Edwards, 1971). Vinylidene chloride-vinyl chloride copolymer resins are alsn extruded into tubes, nods, pipes, and pipe liners for use in contact with chemicals and ether corrosive media fWessling & Edwards, 1971). The multi-layered film of polyethylene an a core of vinylidene chloridevinyl chloride copolymer reportedly has found use in ostary devices (Roth, 1976). In Japan, approximately 8% of the vinylidene chloride-vinyl chloride copolymers produced are in the fom of fibres used far fishing nets, interior furnishings, and construction appliances. Approximately 75% are used in film applications, 13% in latex applications, and 4% in other unspecified applications. The US Pood and Drug administration permits the use of vinylidene chloride copolymers (including the copolymers with vinyl chloride) as components of the following products when they are intended for use in contact with food: (1) adhesives; (2) resinous and polymer coatings; (3) paper and paperboard; (4) rigid and semirigid acrylic and modified acrylic plastics; (5) polyethylene phthalate polymers; and (6) packag ing material for use during the irradiaticn of prepackaged feeds (US Rood and Drug Administration, 1977). CONFIDENTIAL Subject ", ' * Boss v ' ' 17 2.2 Occurrence Vinylidene chloride-vinyl chloride ccpolymers are not known to occur as natural products. 2.3 Analysis The analytical chsnistry of vinyl polymers, including vinylidene chloride-vinyl chloride copolymers has been reviewed (Cobler si si., 1963). Methods of detecting surface finishing agents, such as vinylidene chloride polymers, on paper have been reviewed (Proksch, 1969). Methods of identifying polymer films, including vinylidene chloride copolymers, based an physical and chemical properties have been described (Sriston, 1974; Van Giesan, 1969). 3. Biological Cata Relevant to the Evaluation of Carcinogenic Bisk to Humans 3.1 Carcinogenicity and related studies in animals No data were available to the Working Group. 3.2 Other relevant biological data (a) Experimental systems Rats fed a diet containing 5% jvinyl chlorid^vinylidene chloride/ aopolyrrer for two years showed no toxic effects. Two dogs fed a diet containing 5% of the aopolyrrer were also without evidence of toxic effects (Wilson * McCormick, 1954) - T Rabbits treated for three months i.v. with 1 ml/kg bw 1% solution of vinylidene chloride-vinyl chloride copolymer exhibited hypertrophy of the reticulo-andothellal cells of the spleen, bone marrow, liver, lymphatic tissue and lungs (Miyasaki, 1959). (b) Humans Cne case of contact dermatitis, limited to the area of its application, has been reported from the use of Saran Wrap (a vinylidene chloride-vinyl chloride copolymer). A positive patch test to the copolymer was also obtained (Osbourn, 1964). CONFIDENTIAL Sub`s' t : Rc55 v - - - 3 1- " c~: ' ln , ;-0 - i337 - irt . 1 . Jiina CMA 015632 Two workers developed persistent cranial nerve disorders after cleaning-out tank cars in which an aqueous dispersion of vinylidene chloride copolymers had been transported- These were attributed to trace reaction products found in the copolymer (i.e., nonochloroacetylene and/or dichloroacetylene)- The trigeminal nerve was principally involved, and to a lesser degree the occipital auricular and cervical cutaneous nerves as well as the muscles of mastication, the eye muscles and the hypoglossus (Henschler et at., 1970). 3.3 Case reports and epidemiological studies No data were available to the Working Group. 4. Summary of Data Reported and Evaluation 4.1 Experimental data Vinylidene chloride has been tested by oral administration in rats and by inhalation exposure in mlce^ rats and hamsters. When given by inhalation in rats and mice, it induced malignant tumours, including angio sarcomas. The preliminary results of another inhalation study in rats and mice indicate the induction of malignant tumours of the kidney in mice, mostly males, and of an increased incidence of mammary tumours in rats. No carcinogenic effect was observed in an on-going study in hamsters exposed to vinylidene chloride by inhalation. The oral study in rats is still underway and cannot be evaluated. Vinylidene chloride is mutagenic. JJo data on the carcinogenicity of vinylidene chloride-vinyl chloride copolymer were available to the Working Group. 4.2 Human data The production volume of vinylidene chloride is high, and the material is .utilized almost entirely In the production of the copolymers. This suggests that occupationally exposed groups might be identified for epidemiological investigation. Production of the vinylidene chloridevinyl chloride copolymer is extensive, and its use in consumer products (including food packaging) indicates the possibility of widespread human exposure. CONFIDENTIAL Ross v 14th . . ' Calcasieu Parish, :i Order in '.:o 10*4337 - '`' irt L-s-siana CMA 015433 The only epidemiological study available to the Working Group reported no tumours associated with exposure to vinylidene chloride, but the data were not adequate to permit an assessment of carcinogenicicy. No case reports or epidemiological studies relevant to the carcino genicity of vinylidene chloride-vinyl chloride copolymers were available to the Working Group. 4.3 Evaluation The single epidemiological study on vinylidene chloride available to the Working Group was not adequate to permit an assesst*nt of human carcinogenicicy. In view of the substantial volume of vinylidene chloride manufactured, the use of this compound in copolymers, the identification of the material in Industrial emissions and in drinking water, and its presence in some trichloroethylene, some chloroprena, and in household materials, the lack of human epidemiological studies is serious. The available experimental evidence of carcinogenicity of vinylidene chloride indicates that it produces malignant tumours in mice and rats and that some of the tumours are similar to those produced by vinyl chloride. This evidence is, however, limited by the fact that it is partly based on studies which were still in progress. An evaluation of the carcinogenicity of vinylidene chloride will be made upon completion of studies known to be underway. flOffFlD'BffTl AL Subject x.. Pro -estive 0; der In Boss v. 90-4"37 14th J-l ' Calcasieu 1 -- -- " ----- u - _.r.a CM 015634 L/> * --v 'j. Q= 1 * ^ 5. Peferances - ---^a ^ *-3 ACG3 (1976) TLVs Threshold Limit Values for Chemical Substances in Vfarktccn Air Adopted by ACC-IH, Cincinnati, Ohio, American Conference of Governmental Industrial Hygienists, n. 30 Aleman, ?.D. & Dittrer, D.S. (1966) Environmental 3iology, 3etiasda, Maryland, Federation of American Societies for Environmental Biology, P?. 326, 323 Anm. (1972) Fire Protection Guide cn Hazardous Materials, 4th ed., Boston, -Nan-cnal^Fire Protection Association^ o. 42-229 li t r'__ t ? -> ' ) '* 1 Anon. (1977) Vinylidene chloride linked to cancer. Chem. Ena. News, February 23, to. 6-7 * Andersen, M.S. & Jenkins, L.J., Jr (1977) Qihancsnent of 1,1-dichloroethylene hepatotaxicity by pretreatinent with low molecular weight epoxides (Abstract No. 41). Tbxicol. appl. ?h*rm*^ni., 41, 143 y Anderson, D., Hodge, M.C.2. & Purchase, I.F.H. (1973) Dominant lethal studies with the halogenated olfins vinyl chloride and vinylidene dichloride in male Ol mice. Environ. Health Perspect., 21, 71-78 BartschT^H., Malaveille, C,, Mcntesano, R. & Ttmatis, L. (1975) Tissue- mediated mutagenicity of vinylidene chloride and 2-chlorobutadiere in Salmonella typhzmurium. Nature (Lend.) , 255, 641-643 Bartsch, H., Malaveille, C. & Montesano, R. (1976) The predictive value of tissue-mediated mutagenicity assay to a33ess the'carcinogenic nsk of chanicals" In: Montesano, k., Bartsch, H. & Ttnatis, X., ecs. Screening Tests in Chemical Carcinogenesis, Lyon, IARC Scientific Publications No. 12, pp. 467-491 Birkel, T.J., Poach, J.A.G. & Sphon, J.A. (1977) Determination of vinyLi dene chloride in saran films by electron capture gas-solid chromato graphy and confirmation by mass soectrcretry. J. Assoc, off. anal. Chem., 60, 1210-1213 Boliini, M., Seves, A. & Focher, B. (1974) Determination of free monoiers in water mail signs of synthetic polymers and copolymers. Ind. Carta, 12, 234-240 [Chem. Abstr., 81, 121672b] Briston, J.H. (1974) Appendix 3. Identification of film mat^r-iaig. in: Plastics Films, New York, John Wiley and Sans, pp. 28^-293 Car.Ison, G.P. & Fuller, G.C. (1972) Interaction of modifiers of hepatic microsomal drug metabolism and the inhalation toxicity of 1,1-dichloro- ethylene. Pes. Conn. Chem. Pathol. Pharmaool,, , 553-559 .C ^ * w*. C_ t frC,- . 1st >* ^3:^. * i -- * _ ,, SjQ fehowicai '"arhrioiefy ' VQl. VI, New York, Barnes & Noble Books, ' " pp. 540-543 J / CMA 01563 ~ J- v / Cobler, J.G., Dang, M.W. & Owens, E.G. (1963) Analytical chemistry of vinyl form-forming polymers. Sci. Technol. Polvm. Films, 1, 703-S12 [Chen, jbstr., 70, 73537m] 2A (1376) Health and Environmental Iroacts. Task 1. Vinylidene Chloride, --A-550/6--6-023, Wasrungccn DC) L*S rjrvii'orsna.n-a 1, if-ar~-inn nam, , ea*, Lcraivtrerizarion, New Yorfc, Intarscienca Publisters, p. 609 Gage, J.C. (1970) The subacute inhalation toxicity of 109 industrial chemicals. Brit. J. ind. .Med., 27, 1-13 GrasselLi, J.G. & Ritchey, W.M. (1975) <3C Atlas of Spectral Data and Physical Constants for Organic Carcounds, ind ed.~, Voi. ttt, Chip, Chemical Rubber Co., p. 231 Greim, H., Bonse, G., Radwan, 2., Reichert, D. & Henschler, D. (1975) Mutagenicity in vitro and potential carcinogenicity of chlorinated ethylenes as a function of netabolic cxirane formation. 3iocbem. Pharmacol., 24, 2013-2017 Gronsberg, E.S. (1975) Detennination of vinylidene chloride in the air (Russ.). Gig, i Sanit., 1_, 77-79 Haley, T.J. (1975) Vinylidene chloride: A review of the literature. din. Ttaxiool., 3_, 633-643 Hardie, D.W.F. (1964) Chlorocarbons and chlorohydrocarbons. Didiloroethyler.es. In: Kirk. R.E. & Qtnmer, n.v~, *rlg, pvirryr-1i-!rofn n-4 Chemical Technology, 2nd ed., Vol. 5, New York, John Wiley'ard Sons, op. 178-lSiJ ienschler, D., Broser, F. & Hopf, H.C. (1970) "Polyneuritis cranialis" durdi Vejigiftung mit chlorierten Acetylenen beim Umgang mit Vinylidenchlorid-Oopolymeren ("Polyneuritis cranialis" following poiscring with chlorinated acetylenes while handling vinylidene copolymers). Arch. Ttodkol., 26, 62-75 Jenschlex, D. (1973) Metabolism and rrutageiicity of halogenated olefins. A acnparisen of structure and activity. Environ. Health Persoect., 21, 61-64 ` Husfaan, J. & Homreich, M. (1976) Air Pollution Assessment of Vinylidene Chloride, Springfield, Virginia^ US National Technical Information Service, FB 256 738, p. 40 COfl'FIflBNTI AL Sub;ect to Protects Rcss v : ~ ' -l CMA 015636 14th j ^ ch 7 ' -1 :i t ' , -. Caloasirtu h r io ii, IARC (1978) Information Bulletin on the Survey of Clericals_3eing Tested for Caroruiaaenicity, No. 7, Lyon, International Agency for Pes^artH** cn Cancer, pp. 70 / 257, 272, 278 . ! Irish, D.D. (1963) Aliphatic Halogenatad Hydrocarbons. Li: Patty, F.A., ed., Industrial Hygiene and Tcaacoloc/, Vol. II, 2nd revised ed., '5ew York, Interscience, op. 1305-1307 Jaeger, R.J. (1975) Vinyl chloride ircnaner: canrents on its hepato- toxicity and interaction with 1,1-dichloroethylane. Ann. N.Y. Acad. Sci., 246, 150-151 Jaeger, R.J., Conolly, R.B. & Murphy, S.D. (1973a) Diurnal variation of hepatic glutathione concentration and its correlation with 1,1- dichloroethylene inhalation toxicity in rats. Res. Corn, chan. Pathol. Pharmacol., 6_, 465-471 Jaeger, R.J., Trabulus, M.J. & Murphy, S.D. (1973b) The interaction of adrenalectary, partial adrenal replacement therapy, and starvation with hepatotoxicity and lethality of 1,1-dichloroethylane intoxication (Abstract No. 133). TQxicol. appl. Phasiaool., 25 , 491 Jaeger, R.J., Trabulus, M.J. & Micphy, S.D. (1972) Biochemical effects of 1,1-dichloroethylene in rats: ooncarison with carbon tetra- chloride and 1,2-dichlcroethylene. Tbadool. arol. Pharnacol., 4* 2. "b Sat.Slo " jaeger, R.J., Conolly, R.B. & Murphy, S.D. (1974) Effect of 18 hr fast and glutathione depletion of 1,1-dichlorcethylore-induced bepatotoxicity and lethality in rats. Exp, mol. Pathol., 20, 187-198 Jenkins, L.J., Trabulus, M.J. & Murphy, S.D. (1972) Biochemical effects of 1,1-dichlaroethylene in rats: oompariscn with carbon tetra chloride and 1,2-dichloroethylens. Tcxicol. arol. Pharmacol., 23, 501-510 Jones, B.K. i Hathway, D.E. (1978a) Differences in metabolism of vinylidene chloride between mice and rats. Br. J. Cancer, 37, 411-417 Jones, B.K. & Hathway, D.E. (1978b) The biological fate of vinylidene chloride in rats. Cbern.-biol. Interact., 20 , 27-41 Kiezel, L., Liszka, M. & Rutkowski, M. (1975) Gas chztoatographic deterruination of trace inpurities in distillates of vinyl chloride rrurxner (P61.). <2t. Anal. (Warsaw),. 20, 555-562 [Orem. Abstr., 83^, 212233s] Kramer, C.G. & textchler, J.E. (1972) TSe correlation of clinical and environmental measurements for workers exposed to vinyl chloride. Am. ind. Hyg. Assoc. J., 33, 19-30 | CMA 015637 } J M 'w 4 tu e.J) $r>Q -- \ *"7 ^'V ----------------- -V 7^ ^ i'u "Vj : a ^ 1 W.1 k* ** "T c_\\ { , ^, .. < * w s i - 4-*r Kurginyan, K.A. i Shirinyan, V.T. (1969) Identification and quantitative determination of same impurities in chloroorene. Arm. Khim. Zh., 22, 61-65 [Chem. Abstr., 71, 29974x] ~ lee, C.C., Bhandari, J.C., Winston, J.Mi,., House, W.B., Peters, P.J., Dixon, R.l. & Wbods, J.S. (1977) Inhalation toxicity of vinyl chloride and vinylidene chloride. Ehviron. Health Perspect., 21, 25-32 Ma.lt.oni, C. (1977) Recent findings on the carcinogenicity of chlorinated olefins. Brviron. Health Perspect., 21, 1-5 Maltsni, C., Cotci, G., Morisi, L. & Qiiect, P. (1977) Carcinogenicity bioassays of vinylidene chloride. Research plan and early results. Msd. Lavoie, 68, 241-262 Miyasaki, K. (1959) Experimental studies on the reticulcrendcthelial system by intravenous injection of high molecular synthetic vinyl caipounds in rabbits. Acta Pathol. Jpn., 9, 109-131 Norris, J.M. (1977) Tfaxicoloclcal and pharmacokinetic studies on inhaled and incested vinvLidene chloride ip i^ir?ratorv`ahuTaiJ'.'" gTS36flgad at the 1977 73PPI Paper Synthetics Conference in Cfucago, Illinois, September 26, 1977 Osbourn, R.A. (1964) Contact dermatitis caused by Saran Wrap. J. Am. med. Assoc., 188,`1159 Ott, M.G., Langner, R.R. & Solder, 3.S. (1975) Vinyl chloride exposure in a controlled industrial environment. A long-term mortality experience in 594 employees. Arch, environ. Health, 30, 333-339 Ott, M.G., Fishbedc, W.A., Tbwisend, J.C. & Schneider, E.J. (1976) A health study of enployees exposed to vinyLidene chloride. J. occuo. Med., 13, 735-738 " Perry, R.H. & Chilton, C.H. (1973) Chemical Engineers' Handbook, 5th ed., cNew York, McGraw-Hill Book Co., p. 3-61 ' PPG Industries (1975) Vinylidene chloride, Bulletin 120A, Pittsburgh, Ehiladelphia Prendergast, J.A., Jones, R.A., Jenkins, L.J., Jr & Siegel, J. (1967) Effects on experimental animals of long-term inhalation of trichloroethylene, carbon tetrachloride, 1,1,1-trichloroethane, dichlorodifluortrethane, and 1,1-dichloroethylene. Toxicol, apol. Phantacol., 10, 270-289 Prcksch, A. (1969) Determinaticn of chemical auxiliary and finishing agents [for paper] by rapid methods (Gar.). Allg. Pap.-Rmdsch., 35, 1207-1208 [Chan. Abstr., 72, 4461k] I c-> (f 1-i - CMA 015639 Ranpy, I*-W., Quast, J.F., Haniston, C.G., 3almer, M.P. & Schwetz, 3-A. (1977) Interim results of two-year toxicological studies in rats of vinylidene chloride incorporated in the drinking water or administered by reseated inhalation. Environ. Health Perspect., 21, 33-43 Peinhardt, R.C. (1943) Vinylidene chloride polvmers. Ltd. Ehg. Chen., 35_, 442-448 * ----------- ------------- Reynolds, E.S., Moslen, M.T., Szabo, S., Jaeger, R.J. & Murphy, S.D. (1975) Hepatotcodcity of vinyl chloride and 1,1-dichlaroethylene. Am. J. Pathol., 31, 219-236 ---------- Stoth, s.F. (1976) Saran _coatir.gs - latex or lacquer? In: Chemical Marketing and Sconcnucs Reprints, Staten IsiandT New York7""cSeinical Marketing and Economics Division of the American Chsmical Society, oc. 29-36 Russell, j.w. (1975) Analysis of air pollutants using sampling gas chromatography, Biviron. Sci. Technol., 9_, 1175-1178 and Sassu, G.M., Zilio-Grandi, F. & Conte, A. (1968) Gas-chromatographic determination of impurities in vinyl chloride. J. Chromatoqr., 34, 394-398 [Chem. Afcstr,, 69, 8297x] Severs, L.W. & Skory, L.K. (1975) Monitoring personnel exposure to vinyl chloride, vinylidene chloride and methyl chloride in an industrial vrerk environment. Am. ind. Hvg. Assoc. J., 39, 669-676 Siegel, J., Jones, R.A., Cbcm, R.A. & Lyon, J.P. (1971) Effects on experimental animals of acute, repeated and continuous inhalation exposure?to dichloroacetylene mixtures. Ttodcol. aool. Pharmacol., 18, 168-^174 " Shackelford, W.M. & Keith, L.H. (1976) Frequency of Organic Cgrpounds Identified in tetter. EPA-600/4-76-062Athens, Georgy, US Environ- nsntal ProtectionAgency, pp. 130, 133-^fc5S <2+ ia. US Ehvirannsntal Protection Agency (1975) Preliminary Assessment of Suspected Carcinogens in Drinking Water, Washington DC, p. II-3 . O la -i.. oi *^. ? h, US Food and Drug Administration (1977) '"F&Sd laid LSrugA .I'l'ltLS zi, frarts 175.105, 175.300, 175.320, 175.360, 17^.365, 176:17b, 176.180, 177.1010, 177.1630, 178.3790, 179.45, po. 438, 446, 452, 455, 465, 467-469, 471, 482, 486, 489, 496, 530, 596, 607-608 US Tariff Caitnissicn (1941) Synthetic Organic Chemicals, US Production and Salas, 1940, Report No. 148, Second Series," Washington tc, US Government Printing Office, p. 58 r* V SubJact_ro ?; ' BOSS V 14th Calcasieu Pa ; = ortter in , ho . 90-4337 )- ' 11 ? ; " art isiana CMA 015639 Van Gieson, ?. (1969) Here's a quick, easy way to identify films. Package Big., 14, 76-77 [Chea, Abstr,, 71, 71274u] Viola, ?.L. & Caputo, A. (1977) carcir.ogerJ.city studies on vinylidene chloride. Environ. Health Perspect., 21, 45-47 Vlasov, s.M. & Hodyagin, G.N. (1970) Gas chromatographic analysis of trichloroethylene (Puss.). Tr. Khim. Khini. lekhnol., 1, 161-162 [Chem. Abstr., 75, 71124c] " *>m. *<*. Wilson, R.H. & McConnick, W.E. (1954) ^toxicology of plasticsL. Plastaners and monarers. Ind. Med. Surg., 23, 479-486 * t Windholz, M., ed. (1976) Ove Merck Index, 9th ed., Rahway, N.J., Merck & Go., p. 1283 A L* OaK ^ 1^4 j "Tie i* Uj_ 0l_ Ga-ut." rU._k i to CMA 015640 Page 2 D^AAT 1. Introduction The present report deals primarily with the results of a project of integrated research designed to study the long-term effects (toxic and carcinogenic) of vinylidene chloride (VDC). The project included several experiments, in which the monomer was administered by different routes (inhalation and ingestion) at different concentrations, on three animal species (Sprague-Dawley rats, Swiss mice and Chine hamsters). To our knowledge, this project, sponsored by several European producers of vinylidene choride --European Study Group for VDC Toxicity (BASF AG, Dow Chemical Europe SA, I.C.I. Ltd, and Solvay et Cie) - represents the most comprehensive study available in this area. Moreover, in this report available studies on long-term carcinogenicity bioassays, as well as on other biological parameters concerning the behaviour and effects of VDC, performed in other laboratories and in ours, are reviewed for a better understanding of the biological action of the monomer. .2 Vinylidene chloride Vinylidene chloride (VDC), or 1,1-dichloroethylene, Q^CClj* is produced in the (J.S.A. , Japan, and several Western European countries. Vinylidene chloride is mainly employed for the production of poly(vinylidene chloride) (PVDC) resins and disper sions. Examples are Diofan (BASF), Ixan (Solvay), Saran (Dow), and Viclan (ICI). The most important property of PVDC is its low permeability to oxygen and moisture. CONFIDENTIAL Subject ^ Ross v T ; .; 1* V Caloas:e1 ' "' In : :'3. 90-4837 ' strict Court itish, Louisiana CMA 013641 Page 3 The major use of PVDC is in the manufacture of flexible packaging for food. The following population groups may be potentially expos ed to VDC: (1) workers engaged in the production of VDC monomers; (2) workers manufacturing and processing VDC polymers on an industrial scale; (3) residents around factories producing VDC and VDC polymers; and, to a much lesser extent, (4) poeple undergoing contact with resins made with VDC and with materials containing these resins and (5) consumers of food preserved in packaging materials coated with VDC resins. 3. History Up to 1974 no data were available on long-term effects of VDC. In October 1974, at the XIth International Cancer Congress, held in Florence, Viola reported that he had observed cases of abdominal reticulosarcomas in a group of Wistar rats exposed to VDC by inhalation 4 hours daily, 5 days weekly, at 200 ppm for 5 months and subsequently at 100 ppm for 7 months. At the "Conference on Comparative Metabolism and Toxicity of Vinyl Chloride Related Com) * pounds" held at the National Cancer Institute in 3ethesda in May 1977. Viola and Caputo (1977) reported in detail the results of this experiment and moreover the results of a further experiment in which VDC was tested on Sprague-Dawley rats, at 100 ppm and 75 ppm, under the same experimental conditions. The authors concluded: "A final statement of the pathological results must await the completion of the microscopic examination of the tissues and organs of all animals. Never theless, it seems clear that there is no grossly observable correlation between tumour formation and VDC inhalation." CONFIDENT! A.L Subject- :o . v 1 :-*.*? in Ross 7. 2-S-2!- -- ::-4337 l4"r. ' : Tourt Cq] - ? _ >__ Louisiana CMA 015642 On the basis of Viola's communication in 1974 The Eu ropean producers of vinylidene chloride asked our In stitute and Tumour Centre to undertake the present pro ject of long-term carcinogenicity bioassays on VDC, which was started in the spring of 1975. From the start of the experiment it was immediately apparent that mice were more sensitive to the acute effects of VDC than rats, and male more than female mice (Maltoni et al., 1975). At the beginning of 1977, in a series of reports (at the Accademia dei Lincei, Rome; at the International PVDC Seminar of TAPPI, Hamburg; at the Americn Cancer Society, New York; and at International Agency for Research on Cancer, Lyons), we reported that under our experimental conditions VDC produced kidney adenocarcinomas in male Swiss mice. These early results were then published in full (Maltoni et al., 1977 a, b). In 1977 the interim results of a two-year toxicological study on the effects on rats of vinylidene chloride incorporated in their drinking water or administered, by repeated inhalation, performec at the Toxicology Research Laboratory of Dow Chemical (Midland, USA), were given (Norris, 1977; Ramply et al., 1977). The research plan and results of the Dow studies were summarized as follows: "Male and female Sprague-Dawley rats were exposed to vinylidene chloride (VDC) orally or by inhalation in two-year toxicological studies. VDC was given in the drinking water at mean - S.D. concentrations of zero, 68 - 13 ppm, 106 - 22 ppm, and 220 - 35 ppm, which produced mean - S.D. dosage levels of zero, 5.9 - 0.6 mg/kg, 10.0 * 1.2 mg/kg, and 19.3 - 2.7 mg/kg for male rats and zero, 7.5 - 0.4 mg/kg, ' ' r. r in ) - -,337 - > _irt JjQu.3.3 l3na CMA 015643 3 age r- 12.6 - 1.1 mg/kg, and 25.6 - 2.4 mg/kg for female rats. Forty-eight rats/sex/VDC level and 80 rats/sex in the control group were used in the two year study with an interim kill of an additional 10 rats/sex/ level at 90 days. In the inhalation study, rats were exposed to 0, 10 or 40 ppm of VDC vapour 6 h/day, 5 days/week for 5 weeks, after which the exposure levels were changed to 0, 25, and 75 ppm of VDC. Exposure continued for a total of 13 months and the rats held for observation and additional 6 months. Interim kills occurred at 1, 6 and 12 months. A separate 90-day study using 20 rats/sex/level was conducted at 0, 25, and 75 ppm of VDC vapor. There were 86 rats/sex/level in the two-year portion of the sutdy. The parameters monitored were; body weight, food and water consumption (drinking water study only), haematology, clinical chemistries, cytogenetics of bone marrow cells (inhalation study only), mortality, terminal organ weights, and gross and histopathology. Based .on-interim.kills and.gross pathologic observations, the main conclusions are: increased dytoplasmic vacuolation of hepatocytes was seen in the livers of rats given 200 ppm VDC in drinking water or 25 or 75 ppm VDC vapor by inhala tion; based on gross tumor count, tumour incidence in VDC-exposed rats was not greater than controls." In 1977, in two similar articles, Lee et al. (1977 a, b) reported the results of a study, supported by the Natio nal Institute of Environmental Health Sciences (USA), on the carcinogenicity of vinyl chloride and VDC in Albino CD-I mice and CD rats. In this project, mice and rats of both sexes were exposed by inhalation to 55 ppm of VDC, 6 hours daily, 5 days weekly. Similar groups of animals were kept as controls. The study lasted 12 months. In this experiment the authors reported that they had ob- CC^FIP. 7 :t TI AD in Subject ..; -1337 BOSS-'7' :.c-- 14th -- . * Calcar r - CMA 015644 Toxicity and carcinogenicity bioassays of vinylidene chloride, II. Chronic toxicity and carcinogenicity. by C. Maltoni, G. Cotti, L. Morisi, and P. Chieco Institute of Oncology and Tumor Centre, Bologna, Italy c CV ` OHFIiEyJSK-TkIAX 50 - 4837 CMA 0154-45 Page 6 served 6 lung adenomas among 35 treated male mice versus 1 among 26 controls, 3 liver angiosarcomas (2 in males and 1 in females) and 3 hepatomas (2 in males and 1 in females) from 70 treated mice, versus none from 62 con trols. Two haemangiosarcomas (1 of the mesenteric nodes, l and 1 in subcutaneous tissue) were found in two of 71 treated rats versus none of the 70 controls. Concerning the bronchiolar adenomas and the hepatomas in mice, the authors concluded that their significance is questionable, since the tumours may accur also in untreated animals (and probably in consideration of the small number of tested animals). As far as angiosarcomas are concerned, on the basis of our experience with long-term effects of VDC, this result was unexpected by us (Maltoni ec al.f 1977 a, b). The possibility of unwanted exposure to vinyl chloride used in parallel experiments was discussed at Bethesda "Conference on Comparative Metabolism and Toxi city of Vinyl Chloride Related Compounds" (1977). The compatibility of a prolonged inhalatory exposure of mice to 55 ppm of VDC with a long survival does not parallel our experience on the acute toxicity in several strains of mice (Swiss, Balb/C, C3H, C57 Black), and on the chronic toxicity in Swiss mice, which appeared to bw much more sensitive to toxic effects of the monomer. 4. Planning, materials, and methods The chambers for inhalation exposure were basically built of stainless steel and glass. The determination of VDC concentrations was carried out by gas chromatography. For the ingestion experiments VDC was administered in olive oil by gavage with a stainless steel stomach tube. Subtfec Boss v 14th CalCaJ-^ parish, Louisiana CMA 01564& VDC was supplied and analyzed by the Central Laboratory of Solvay (Brussels). The impurities and their levels in the VDC employed were as follows: 1.1- dichloroethylene (VDC) 1.2- trans-dichloroethylene acetone methylene chloride mono and dichloroacetylene 999.5 g/kg 0.4 0.1 0.05 " 0.02 '* As stabilizer -methoxyphenol at 200 ppm was used. The experiments utilized Sprague-Dawley rats, Swiss mice and Chinese hamsters. All the animals, except the ham sters, had been bred in our Institute for years, and, whatever their use, were all examined at death by complete autopsy, giving us extensive information concerning their current pathology. .The animals were weaned, and classified by sex when 4-5 weeks old, at which time they were numbered by ear punch and divided into groups by litter distribution. After weaning, the animals were given drinking water ad libitum and an adequate commercial diet. The animals were kept in groups of five in Makrolon* cages with tops made of stainless steel wire, but during the period of inhalatory treatment they were housed in groups of ten in stainless steel wire cages with a solid bottom of the same metal. A shallow layer of white-wood shavings served as bedding. The animals were kept in a temperature-controlled labora tory at 19-20 C. Five experiments were started around the same period. The plan of the experiments is given in tables 1 to 5. *Polycarbonate Experiment 1 (BT401) studied the effects on rats of inhalation exposure to 200 (then reduced, after two exposures, to 150 because of the strong coxic effects), 100, 50, 25, and 10 ppm of VDC, 4 hours daily, 4-5 days weekly, for 12 months. One group of animals served as control. 150 ppm was the highest tolerable dose by Sprague-Dawley rats for long-term exposure. From time to time the treatment was reduced from 5 to 4 times weekly because of early toxic effects in animals exposed to 150 ppm. Experiment 2 (BT402) started with groups 1, II, III, IV, V, and VI, and it was planned to study the effects on mice of inhalation exposure to the same range of VDCdoses as BT401. 25 ppm was found to be the highest tolerable dose by Swiss mice for long-term exposure. Doses of 200 and 100 ppm for 2 days, and of 50 ppm for 4 days (4 hours daily) caused death of a proportion of animals. When it appeared that exposure to 200, 100, and 50 ppm had to be withdrawn because of high mortality and severe toxic effects, the number of animals exposed to 25 ppm was enlarged by adding a supplementary group (IV/2), with its proper controls (group VII). In conclusion only two dose levels (10 and 25 ppm) were selected for this long-term study. Experiment 3 (BT403) was designed to evaluate the effects of VDC on Sprague-Dawley rats by ingestion, at three dose levels: 20, 10, and 5 mg/kg body weight, once daily, 4-5 days weekly, for 52 weeks. Experiment 4 (BT 404) was started to complement experimen BT403 with a lower dose (0.5 mg/kg body weight) once daily, 4-5 days weekly, for 52 weeks. CMA 015648 Page 9 --- & ^ ^ Experiment 5 (BOT405) was included to study the effects of VDC in Chinese hamsters at 25 ppm, which is the highest possible concentration for long-term exposure of mice and also one of the exposure levels of rats. The animals were examined weekly, and weighed every 2 weeks during the period of treatment and monthly after the treatment was over. All the detectable gross patho logical changes were recorded during the examination. All the animals were kept under observation until spontaneous death. Moribund animals were isolated, in order to avoid cannibalism. A complete autopsy was carried out on each animal. Histo logical examinations were performed on the Zymbal glands, incerscapular brown fat, salivary glands, tongue, lungs, liver, kidneys, spleen, stomach, different segments of the intestine, bladder, brain, bone marrow (sternum) and any ocher organs with pathological lesions. Furthermore, cytological examinations were carried out on the bone ' marrow of the femur. Histological' sections were stained routinely with hematoxylin and eosin and, in particular cases, with Papanicolaou, Van Gieson, Mallory, and Gomory stains, silver stain, and Congo red. Cytological smears were stained with Giemsa and Papanicolaou stains. O Part of the results have been evaluated by statistical analysis (see Appendix). 5. Results 5.1. Chronic toxicity The toxic effect was studied by histological examination of tissues and organs in animals that had died spontaneous ly. Therefore the observed changes had to be considered Boss v ___ i Louisiana. "Calcasieu Parian. CMA 015649 -a ge J as partly due to the treatment and partly due to spon taneous pathology and to immediate causes of death. 5.1.1. Rats ; i I Regressive changes, such as hepatocytes vacuolisation, cloudy swelling, fatty degeneration (Fig. 1), necro biosis, and necrosis were found in some animals, in treated as well as in control groups. These changes were observed more frequently in rats exposed to 200-150 ppm by inhalation (57.6 %) than in controls (20.5 %). 5.1.2. Mice Pathological changes were found in the liver and in the kidneys, both of control and Created animals. The most frequent findings were regressive changes (hepa tocytes and vacuolisation, cloudy swelling, fatty degene ration, necrobiosis and necrosis) and amyloidosis in the liver, and regressive changes (cloudy swelling and necro sis of tubular cells), amyloidosis of glomeruli, and chronic nephritis in the kidneys. The incidence of these ) * lesions in the groups of animals treated with doses compatible with long survival (25 and 10 ppm) and con trols is given in Tables 6 and 7. From these results no correlation emerges between the abovementioned changes and exposure to VDC. A higher incidence of more pronounced regressive or phlogistic changes was found in residual parenchyma of kidneys with renal adenocarcinoma. In this situation it is not clear if these changes were due to treatment or to the presence of the tumor. CONFIDENTIAL Subject tc Bess v : J?r in - i 337 Calcasi :-'i I;;.. -I' ---ana CMA 015650 Page 11 Areas of fibrosis, with or without calcium deposits (Fig. 2), were found in Che liver of some of the few animals which survided treatment at the high doses of VDC. Such lesions were not noted in control animals. They should be interpreted as reparative scars, following severe necrosis of hepatic cells. 5.1.3. Hamsters No particular changes were found in hamsters, neither in treated nor in control groups. 5.2. Carcinogenicity 5.2.1. Rats Different kinds of tumours were observed in rats treated by inhalation and ingestion and in the control groups, the mammary ones being the most frequent. The incidence of the different types of neoplasias, and the distribution of the different histotypes of mammary tumours and of some biological parameters related to them, are given in Tables 3-22. Q In rats exposed to VDC by inhalation (Tables 3 & 11), and increase in the incidence of mammary tumours in general, but not of carcinomas, was observed among treated animals when compared with the control. Within the treated groups no- dose/response relationship effect was found. No in crease of mammary tumours was observed among rats to which VDC has been given by*stomach tube (Tables 13, 16, 13, and 21) (see statistical evaluation appendix). This phenomenon has been noted in other experiments studying compounds chemically related and also related to ' : n _ 337 Eoss '-T __ . - * ,"irt 14th * -4'.: Calcasieu Parish, Louisiana CMA 015651 VDC, suggesting that this increase of mammary tumours may be due to some "non-specific factors" connected to inhala tion (such as olfactory stimulation with consequent stimulation of pituitary gland). No other relevant findings have been recorded. 5.2.2. Mice Because of the high early mortality in the groups treated with 200, 100 and 50 ppm, the results are mainly based on the groups created at 25 and 10 ppm and on controls. w Various types of tumours were observed in mice, the most frequent ones being kidney adenocarcinomas, mammary tumours, pulmonary adenomas and leukemias. The incidence of these different types of tumours, and data on the histocypes of mammary neoplasias are given in Tables 23-25. 5.2.2.1. Kidney adenocarcinomas The most important finding was the onset of kidney adenocarcinomas in mice exposed to 25 ppm of VDC. This c kind of tumour was also observed in one of the surfivors of the 50 ppm group. Such tumours wer3~>-found^neither in the 10 ppm group(aoc=tn^the control groups, and they have >>> been noted in untreated Swiss mice of our breeding. V^ *)rl &^ The tumours of the kidneys were almost exclusively in males (29 of Che 30). The kidney adenocarcinomas were found in left and right kidneys. In some cases they were bilateral (Fig. 3). Often in the same kidney they appeared to be multi- i ' \ `J Suu ECS? . 14th J-ii:: - : j37 ........ ;'t Calcasieu Parish,, Louisiana CMA 015632 Page 13 centric. At the autopsy the tumours were often quickly detected as nodules of different sizes. Often they were observed in uniformly enlarged kidneys, and also in kidneys without any peculiar gross change. Following different histological parameters, these tomours could present all the patterns occurring in current human pathology. The three different basis classical histotypes were observed, i.e. clear, granular and dark cell carcinomas in dark cell carcinomas, the dytoplasm could be basophilic or strongly eosinophilic (Fig. 4-12). While some tumours were rather monomorphic, others presented pleomorphism of different degree: large cells with very voluminous nuclei were frequently found (Figs. 13-15). The adenocarcinomas were found to show tubular, papillary, cystopapillary, solid, and uniform arrangements (Figs. 16-18). Several of these different patterns in different combinations could be found in the same tumour. Two of these tumours were found with metastases, one in the lung, and one in the liver. Hyperplasia and dysplasia of tubuli were found in the kidneys of some treated male mice (Fig. 19 & 20). 5.2.2.2. Hammary tumours and lung adenomas O Mammary tumours, almost exclusively carcinomas, were found both in treated and in control groups. They were of the type usually observed in the uncreated animals of our breed of mice. A higher incidence of these tumours was observed in the treated groups, without a dose/response relationship (Tables 23 & 25). * Pulmonary adenomas, some with cellular atypias, were found in treated and in control animals, with a higher incidence in the treated ones, without dose/ response relationship (Table 23). Sut: Boss " 14th ' Calcast- j. Parish, Louisiana CMA 015653 ?age ' Statistical analysis was performed to assess the signi ficance of the differences in the incidence of mammary tumours and pulmonary adenomas in mice exposed to 25 and 10 ppm and in controls, also in relation to sex and age. The important of analyzing these factors emerged from the following considerations: (1) the spontaneous incidence of these tumours varied in the two sexes; (2) these tumours were age-correlated, as are many other spontaneous tumours; (3) in experiment BT402 the survival of mice exposed to 25 and 10 ppm was higher than survival of the con trol groups. The data on mortality rates and the results of statisti cal analysis are fully reported in the Appendix. They show: (1) There were some significant higher rates of mammary and pulmonary tumours in VDC-exposed groups when c compared to the controls. <2) When the incidence of these tumours was adjusted for survival rate, the significance of the difference between treated and control groups was reduced. (3) No dose/response relationship could be calculated, either during the total time of the experiment or in different time intervals. (4) Overall there was a clear fluctuation and imbalance of this trend in the different groups. Q '' '" *y Suttee4: - `n Boss v j7 14th J'-, 'i = *: -t- -'-vArz Calcasieu Farish, Louisiana CMA 015A54 Calcasieu Page 15 In conclusion the direct relation of this effect to VDC treatment remains open. 5.2.2.3. Leukemias No correlation between leukemias and VDC treatment was found. 5.2.3. Hamsters Tumours (mainly uterine carcinomas) were found in treated as well as in the control groups. Their incidence was not increased by VDC treatment. 6 6.1. Related research in our and other laboratories Acute toxicity In acute experiments, when VDC was administered at rela tively high doses to rats, it caused changes in the liver and plasma enzymes (Jenkins et al., 1972; J3ger et al., 1972 and 1973). In rats exposed by inhalation to 200 ppm of VDC, J3ger et al. (1975) observed a massive mid-zonal hepatic necrosis with hepatic thrombosis and chromatolysis within 2 h after a 4-h exposure. Serum transaminase and sorbitol dehydrogenase were greatly elevated at 6 h. This effect in unfed rats was associated with glutathione (GSH) depletion. An extensive study of acute toxicity was performed in our Institute (Maltoni et al.* 1975, in press), on SpragueDawley rats and 4 strains of mice (Swiss, Balb/C, C3H, C57 Black). From this study it emerged that mice are more sensitive than rats, and that Swiss and Balb/C mice are ' in CMA 015455 Page 15 r*- more sensitive than C3H and C57 Black mice. Apart from C3H mice, females appeared to be more resistant than males. Among the various effects observed the most pro minent was necrosis, with cariorexis, in liver and in kidney. Such lesions appeared very marked and extensive in Swiss and Balb/C male mice. 6.2. Mutagenic effects Greim et al. (1975) and Bartsch et al. (1975) obtained positive results with the Ames test. These authors point ed out some species-specific differences. VDC given alone showed no mutagenic effects on Salmonella strains. In adding a metabolic system of rat liver enzymes, small mutagenic effects could be seen. Using mouse-liver microsome s , the effects were several times greater. These results parallel certain metabolic differences in the animal species. The ability of monoxygenases to form epoxides is twice as high in mice as in rats. In one of the further metabolic steps the epoxides can be trans formed, by means of epoxide hydratases to diols or other compounds: this enzyme system is seven times more reac tive in rats than in mice (Oesch and Glatt, 1976); there fore, mice seem to produce much higher concentrations of O reactive epoxides or their subsequent products of meta bolism, which are postulated to be the possible direct active toxic components. 6.3. Cytogenetic effects Balmer et al. Dow/MCA, 1975) could not detect any cyto genetic effects in rats exposed 6 weeks by inhalation to VDC in air at concentrations of 25 and 75 ppm. This was confirmed by BASF (1977), where in Chinese hamsters, exposed 5 days by inhalation to 100 ppm VDC, no increase Subject " Ross v, > r. j, 14th 1 * .Calcasieu Pariah, ~ rt Loui; iana CMA 015656 of chromosome aberrations in the bone marrow test could be found. After long-term exposure of 12 months to a concentration of 55 ppm Lee et al. (1973) could not find changes in the cytgenetic analysis of bone marrow test on rats as well as mice. After a single oral dose of 217 mg/kg b.w. of VDC no statistically significant increase of the chromosome aberration rate was found (BASF, 1976). On the basis of these results no cytogenetic effect of VDC could be found, either by long-term or by acute application of a high dose, in inhalation and oral stu dies with rodents. 6.4. Pharmacokinetic and metabolic studies Jaeger et al. (1974) reported that the hepatotoxic effect of VDC is much higher in unfed rats than in fed rats, and that there is an inverse correlation between hepatotoxicity and the concentration of glutathione. These findings led to the hypothesis that the detoxification of VDC is primarily dependent upon the glutathione concentration. Jaeger et al. (1977) pointed out that in unfed rats and C t^ fed rats exposed by inhalation to high doses of C-VDC, 14C radioactivity was found in various organs, particular ly in kidney and in liver. The largest amount of total radioactivity was detected in the kidney of unfed rats. 14C activity in liver cells was detected in mitochondrial, microsomal, and cytoplasmatic fractions. Significantly more radioactivity was found in all liver fractions of unfed rats. The administration of trichloropropane epoxide, an epoxide hydrase inhibitor, seemed to increase the toxicity of VDC significantly in unfed rats, the enhance- C O'M7 QMMTT A-ti ;, - gr in r 90 - 4837 - " i irt . ,'__-iana CMA 015457 Page 13 ment of toxicity being proportional to the concentration of the inhibitor. This last result supports the hypothe sis of the role of epoxide derivatives in toxicity of VDC. McKenna et al. (1977) showed that in rats and mice exposed to 14C-VDC, 14 C was covalently bound to macromolecules in liver and kidney of both species. The j.4 C acti vity was higher in liver and in kidney of mice, in compa rison to rats. The authors concluded that these data indicate an enhanced production of reactive metabolites of VDC in mice. The data of Jaeger et al. and McKenna et al. bring further evidence that VDC toxicity does not correlate directly with the exposure per se, but is reflected in the balance of the amount of toxic metabolites and detoxification processes, which in turn are dependent upon many biologi cal factors. 6.5. Epidemiological studies In a recent study, published in November 1976, Ott et al. examined the mortality and the health control findings among 138 employees exposed to measured levels of VDC, O ranging from 5 ppm to 75 ppm TWA (time weighted average), where vinyl chloride was not used as a second monomer. There were no findings in that study significantly rela ted to VDC. Hepatic damage was observed in 2 individuals with a positive history of ethanol consumption. The authors recommended "that additional epidemiologic data should be developed where exposure has taken place, with careful consideration given to obtaining an accurate history of alcohol consumption". In TTNT I A-Tj Sub'e" loss ; _ U"'" ` _ Caloasl -n r . - In -;37 a CMA 015458 ?age 19 ~2s s'c-' In a similar study Thiess et al. (1977) reported mortali ty rates and health control findings of 629 employees exposed to VDC as well as to vinyl chloride and acrylo nitrile. The VDC working-area concentrations were about 50 ppm (1955-1965), about 10 ppm (1965-1975), and 5-10 ppm after 1975. No significant increases in mortality or tumour incidence were found. Two lung carcinomas occurred at a very young age but with an exposure period of 14 and 25 months only (smoking habits unknown). 7 Conclusions (1) Kidney adenocarcinomas were observed in Swiss (almost exclusively male) mice exposed by inhalation to VDC at 50 and 25 ppm. Under our experimental conditions, 50 ppm VDC was already a near lethal dose, and 25 ppm was the highest dose compatible with long survival of the treated animals. No renal tumours were observed in mice exposed to 10 ppm VDC or in controls. (2) Kidney adenocarcinomas were not observed in rats exposed to VDC by inhalation at doses from 150 to 10 ppm, or by ingestion (gavage) at doses from 20 mg to O 0.5 mg/kg b.w., or in hamsters exposed by inhalation to 25 ppm. (3) The increase of mammary tumours in rats exposed to VDC by inhalation was not dose-related, and there fore it is difficult to evaluate its specific rela tion to the administration of the monomer. This phenomen has been observed in other long-term inha lation bioassays. pr>v S\:v` - ' ' B; 3; ; __ 14th Jaileiil ` Calcasieu Parish, 4.T. - ,r\ ;J37 - ' -art Louisiana CMA 015659 ? age Z> s -- (A) The dependence of the increase of mammary tumours and lung adenomas in mice on VDC treatment still remains open and needs further clarification. (5) In our experiments we did not observe liver angio sarcomas, either in treated rats or in mice. The animals used were of the same breed that is highly susceptible to induction of angiosarcomas by vinyl chloride. (6) Our results have failed to confirm Viola's prelimi nary findings, which suggested that VDC was causing reticulosarcomas in rats. (7) Under our experimental conditions, adenocarcinoma of kidney in mice was the only tumour specifically linked to VDC exposure. (8) __________ ___ __ As far as this effect was concerned, there was a strong host influence: mice were more susceptible than rats and hamsters, and male mice much more than female mice. (9) There was a clear cut direct relationship between the degree of susceptibility to acute toxid and carcinogenic effects in the different animals spe cies and sexes studied. (10) As a result of in vivo and in vitro studies it is probable that VDC is metabolized into more reactive compounds responsible for the toxic, mutagenic and carcinogenic effects of the monomer. The rate at which the accice metabolic compounds are formed and metabolized depends on the concentration of admini stered VDC and on Che metabolic pathways in the tested' animals, which appear to be strongly influenced by species and sex, as well as by other factors. C Subj * ^ Hess v 14tn J -1i -al Calcasieu Parish, 57 Louisiana CMA 015660 (11) The mice used had the appropriate metabolic condi tions for indicating the oncogenic potentiality of VDC. (12) From a scientific point of view, the results of the studies on toxic, mutagenic, and carcinogenic effects of VDC and on its metabolism represent an exceptional integration of experimental multidisciplinary infor mation. (13) We think that further investigations are needed in biological, biochemical, epidemiological, and medi cal fields, in order to (a) further investigate the roles of species, strain, and sex on the pathological effects of VDC; (b) learn more about the metabolism, with particular regard to active metabolic compounds, and the bio logical factors influencing the metabolic pathways; (c) collect all possible epidemiological and medical data on occupationally exposed groups, which to gether with experimental results will provide the background for risk assessment. O 8 Summary The present report deals with the results of a wide array of correlated and integrated experiments aimed at the study of the long-term effects of VDC, with particular regard to carcinogenicity. The monomer is employed in the production of polymers whose major use is in the manu facturing of flexible packaging for food. CONFIDENTIAL Subject vc -- Ross v * ' . 14th CalcasisU j-*---- CMA 015AA1 D age Boss i The monomer was rested by inhalation, in Sprague-Dawley rats (200-150, 100, 50, 25, 10 ppm), Swiss mice (200, 100, 50, 25, 10 ppm), and Chinese hamsters (25 ppm), and by ingestion in rats (at the doses o 20, 10, 5, 0.5 g/kg body weight, in olive oil, by stomach tube). The treatment was carried out 4-5 times weekly, for 1 year, with the exception of the groups of mice, treated by inhalation, at 200, 100 and 50 ppm, in which it was withdrawn after a few days, because of the excessive acute toxicity. 25 ppm is therefore the highest tested dose, compatible with a long-term treatment, by inhala tion in mice. The most important result was onset of kidney adenocar cinomas in mice exposed by inhalation at 50 and 25 ppm. Male mice appeared to be more susceptible than females. This tumour was not observed in mice exposed by inhala tion at 10 ppm VDC or in the other treated animal species. Increase of mammary tumours in rats and mammary tumours and lung adenomas in mice were found following inhalatory exposure to VDC. The lack of dose/ response relationship, the results of statistical analysis, and biological information from these as well as from other correlated experiments leave the problem of the correlation betw en these tumours and inhalation exposure to VDC or other monomers still open. The available data based upon the studies performed in our and other laboratories show good correlation among the results of studies on carcinogenicity, toxicity, mutagenicity, and metabolism of VDC. On the basis of our present knowledge on VDC toxicity and carcinogenicity, the need for further experimental and D3NTIAL CMA 0154*2 ?age 23 epidemiological investigations for risk assessment has been suggested. 9 References Balmer, M.F., Ramply, L.V. , &Quast, J.F. (1975). Dow Chemical Co., unpublished data. Bartsch, H., Malaveille, C., Montesano, R., & Tomatis L. (197 Tissue-mediated mutagenicity in vinylidene chloride and 2-chlorobutadiene in Salmonella thyphimurium. Mature, 225, 641-643. BASF (1976). Unpublished results. Greim, H., Bonse, G. , Radwan, Z., Reichert, D., & Henschler, D. (1975). Mutagenicity in vitro and potential carcinogeni city of chlorinated ethylenes as a function of metabolic oxirane formation. Biochemical Pharmacology. 24, 2013-2017. Jaeger, R.J., Conolly, R.B., & Murphy, S.D. (1974). Effect of 18th fast and glutathione depletion on 1,1-dichloroethylene-induced hepatoxicity and lethality in rats. Experimental Molecular Pathology. 20, 187-198. Jaeger, R.J., Conolly, R.B., Reynolds, E.S., & Sheldon D.(197 Biochemical toxicology of unsaturated halogenaced monomers. Environmental Health Perspectives, 11, 121-128. Jaeger. R.J., Shoner, L.G., & Coffman, L. (1977). 1,1-Dichloroethylene hepatoxicity: proposed mechanism of action and distribution and binding of 14C radio activity following inhalation exposure in rats. Environmental Health Perspectives, 21, 113-119. c_; Sub.,'?'- ' Ross v 14th Calcasieu Paris -1 \ rJ " : t in '-4337 ina CMA 0156&3 Page 24 Jaeger, R.J., Trabulus, M.J., & Murphy, S.D. (1972). Vinylidene chloride hepatoxicity and dissociation from a liperoxidative mechanism. Toxicology and Applied Pharmacology. 22, 310. Jaeger, F.J., Trabulus, M.J., & Murphy, S.D. (1973). Biochemical effects of 1,l-dichloroethylene in rats: dissociation of its hepatoxicity from a lipoperoxidative mechanism. Toxicology and Applied Pharmacology. 24, 457-467. Jenkins, L.H. , Trabulus, M.J., & Murphy, S.D. (1972). Biochemical effects of 1,l-dichloroethylene in rats: comparison with carbon tetrachloride and 1,d-dichloroethylene Toxicology and Applied Pharmacology. 23, 501-510. Lee, C.C., Bhandari, J.C., Winston, J.M., House, W.B., Dixon, R.L., & Woods, J.S. (1978). Carcinogenicity of vinyl chloride and vinylidene chloride. Journal of Toxicology and Environmental Health, 4, 15-30. Lee, C.C., Bhandari, J.C., Winston, J.M., House, W.B., Peters, P.J., Dixon, R.L., & Woods, J.S. (1977). Inhalation toxicity of vinyl chloride and vinylidene chloride. Environmental Health Perspectives, 21, 25-32. Maltoni, C. (1977 b). Recent findings on the carcinogenicity of chlorinated olefins. Environmental Health Perspectives, 21, 1-5. Maltoni, C., Cotti, G., & Chieco, P. (1975). Tossiciti acuta da cloruro di vinilidene: l'effetto della specie e del sesso. Gli Ospedali della Vita, 2/6, 105-106. --1--Subject^ t Boss v 14th Calcasieu Parian, in ;-0 -4837 _j1ana CMA 015664 Page 25 Maltoni, C. , Cocci, G., Morisi, L. & Chieco, P. (1977 a). Carcinogenicity bioassays of vinylidene chloride (research plan and early results). La Medicina del Lavoro, 68, 241-262. Maltoni, G. , Patella, V. , & Maioli, P. (In press). Toxicity and carcinogenicity bioassays of vinylidene chloride: acute toxicity. McKenna, K.J., Watanabe, P.G., & Gehring, P.J. (1977). Pharmacokinetics of vinylidene chloride in rats. Environmental Health Perspectives. 21, 99-105. Morris, J.M. (1977). Investigation of the potential toxicological effects and pharmacokinetics of inhaled and ingested vinylidene chloride in laboratory animals (MCA-Toxicology Programme for vinylidene chloride). Paper presented at European TAPPI Meeting, Hamburg. Oesch, F., & Glatt, H.R. (1976). Screening tests in chemical carcinogenesis. IARC Scientific Publications. 12, 255-274. Ott, M.G. , Fishbeck, W.A.Townsend, J.C., & Scheider, .E.J. (1976). A health study of employees exposed to vinylidene chloride. Journal of Occupation Medicine. 18, 735-738. Rampy, L.W., Quast, J.F., Humiston, C.G., Balmer, M.F., & Schwetz, B.A. (1977). Interim results of two-year toxicological studies in rats of vinylidene chloride incorporated in the drinking water or administered by repeated inhalation. Environmental Health Perspectives. 21, 33-43. Viola, P.L. (1974). Communication at the XI International Cancer Congress, Florence. Unpublished data. flrumPEKTIAL Boss v. Ccmm 14th Cains"--' CMA 015465 Page 26 ^ 's' VUU, P.L., S, Caputo, A. (1977). Carcinogenic!cy studies on vtnylidene chloride. Environmental Health Perse-.......... 21, 45-47. *--------- I o ^roJ:a"tiv&..r*yC'4337 Subject t0 I' ' - * .' .. , ?.0 S 3 _ V - -' 14tb Calcasieu r c... CMA 015444 TAV.^F 1 t-rin;rr:t Si-lOl fx.'c by inh.-ilattcn to Vinyl loana Cill c>ri J i (VOC) ir. air at 100, 30. 2>, 10 tpm, A hours daily, 1-3 d.'.j-.s t.'ckly, lot t2 vs.-td:". A'tlHALS GVO'j?c ' * co;:CKNTR/:riOK* ) (loy rats, 16 veclts old at start) _______________________ l :!f-0 ( i) - 130 mm: 0 9 ____ '"_______ 1 Total I II j 100 !>p:n j -ISO jCr i so ; 1u .......... ............. 1 i 1 to ITT i Vj ` 30 | 30 ' .............. ' i i IV 2:j pm.: 30 30 60 V 10 ppia VI 1 JTotal "? trMt (Control*:) 1 j 1 J 30 < 100 j | 1 230 j 30 ,w 230 r, o 200 I 55C (a) TVo treatrcuts only bsc&uso of the high toxicity of this dose level. r>>so0 111 c oX. & * * Ti mr TABLE 2 i,B''$xJ>^rimcnt BT402 : Exposure by inhalation to Vinyl id one Chloride (VDC) " in air at 200, 100, 50, 25, 10 ppm, 4 hours daily, 't 4-5 days weekly, For 52 weeks. v'.' > *>j g.iciips ;;o. Cone cntiMti on ANIT'.a1.S (Swiss mica 16 weeks old (group I, II, III, IV, V, VI) and 9 week:: old }"'} pp.it l 50 rp.n 1 reeks(d; 30 | i...................... " 30 I 60 | IV 25 ppm 52 weeks 30 30 60 IV bis (a) 25 ppm 52 weeks 120 120 240 V 10 ppi . 52 waeks , 30 30 CO VI ilo treatment (Cciurols.)tb) _ 100 100 200 VII No trea tr.ien t (Controls)(c) 90 90 130 Total 490 490 980 (a) The treatment started two weeks later than in other groups. (b) Controls to the groups I, II, III, IV, V. (*') Controls to the group IV bis. (d) Tr.e treatment was interrupted because of the high toxic effects and high mortality. "w1 CO >0 -0 in < u r 'to-e r. 1 a <> r, 4 o ,, <4 n E-1 fZ5 -SM' f=) H ia)i o ri o o o<pD , i-oH >a ^ s S TABLE 3 Exrerin.ent E74C3 : Exposure by ingestion (stonach tube) to Vinylidtne Chloride (VDC) in olive oil at 20, 10, 5 ^.g'/Kg tody veigiit, onca daily, 4-5 days veelrly, for 52 reexs. 0 VOLTS xc. T ; i: i 1 111 IV Total ' COPCEHTPATICH 20 mg/Kg 10 sir/Kg 5 mg/JCg None (olive oil alone) (Controls) /*!!* '.11*1.1 ("Spragiie-Darley rats, !) rae3:s ole at sea:*!.) r 0* ? Tot?. 1 i j { 50 50 100 50 50 ICO i 50 50 100 too 100 200 250 250 500 CS >0 ID r a r I' J t - nl C O N F ID E N T IA L o0) r^> +O-> Or cfl CO tC9ofD)li Q 1 >1 > 'I, (1 V 0 ( TABLE 4 / V v% siV"r,* c- \or irnon t BT404 : 5r.poi.tvrR by inge'stion (stomach tube) to Vinylidene *Y Chloride (VDC) in olive oil at 0.5 rcs/Kg body weight, 'I wvcs Sjj or.ee daily, 4-5 (lays weekly, for 52 veeks. CROLTS NO. I II Total COltCSNTSV.TICl' c,5 ir.g/Kg Hone (olive oil alone) (Contrcls) A.nuw, (Spragus-Dav;! ;y rats, 10 week a* 5 50 50 old at. start) Total 100 j1 92 . 132 77 127 159 259 >UoT xo. tacffll @ t rr\jI ft- \ TAELE 5 A,;>." s,'^enniGnt 9T405 : Exposure by inhalation to Vinylidcne Chloride (VDC) f*/ ^ c\ Uto' lOlCCHt 3= ,J in sir at 25 ppm, 4 hour3 daily, 4-5 days weekly. for 52 weeks. GROUPS NO. I 11 Total CONCENTRATION 25 Ppm No treatment (Controls) AMIMAI-S {Ch'inese hamsters, 23 weeks old at start) tf1 9 Total 30 30 60 1 18 17 35 48 47 95 I rv o HIO o Xu C alcasieu lir U : . , ^-L -cia n a `o fj 1 r ri-- jn o f'4 - Or ?o OP V * t> to 5 6 7 2 ------ /A / 9 ,\ 1 , ' >0,CCKI Lj*?/] A >'/ TAHLK 6 Experiment DT402 : Exposure by inhalation to Vinylidene Chloride (VDC), in air at 25, 10 ppm., 4 hour3 daily, 4-5 days weekly, for 52 weeks. R.es.u.lt.a aTter 121 woolca (end of experiment). * KON NE0PLA31TC LESIONS OF liv-ti CROUPS no. TREATfGfiT Concentration Length AHIEAL3 (Swiss nice 16 vd.s old(Crs IV, VT.VJ) and 9 w'-at (IV'JitOut nVa-.'t AWIEALS 171TH NON NEOPLASTIC LESIGtS OF LIVES Examined animals Rogrs ftsive changes Amyloidosis ] I ;v 1 r> r\t IV bis 1 j 25 ppn. 1 ( 25 ppm. *t ) V \ 10 ; pr.. Sex No. c.t Mo. start Me. * 'j? __________ 0 V d1 Fc 0 a* .10 rr.O0 91*3 0 JO -W 100.0 ..... r 6.7 cn | \ 60 50 ' 3.3 120 107 89.2 7 6.5 52 reokn 9 ! 20 111 92.5 9 3.1 j Ail _____________\_________ t__ 240 2 i 6 90.8 16 7.3 a" .10 26 86.7 0 - 52 '.veclrr V o'* o ........ t.. a1' 10 60 ICO 27 50.0 53 83.3 85 85.0 3 11.1 3 5.7 5 5.9 Total Mo. * 8 28.fi + i ea) No. ( fi [' 3 | 23.6 ! u> No , | ^ 1 '!- J 1A.0 | i * 11 19.0 11 | 19,0 ! 0 j 27 25.2 \ 22 t j` 20.b j 4 > 39 35.1 | 27 | 24.3 j 12 1,f -0.3 66 30.3 | 49 7 >.5 r i V, A 7 26.9 6 23.1 i 6 22.2 4 1.1.8 'i 3.6 i 13 24.5 73 35.9 10 11 18.9 12.9 3 62 5.7 '2.9 VI |Total Mo treatment (controls) 0+ d& o 4 100 91 91.0 3 3.3 63 69.2 18 19,6 :*y i9.fi 200 176 08.0 8 4.5 136 -77.3 29 16.5 107 60.6 560 5C5 * i i o c r u * * m :r............ ..................T-r iBpt. ut' ; 'J ' ` 1 - l'-'1 kk,jjf 'i nl V'^fqng i ! ^TJO TABLE 7. r* cnours j;c. TfCM'HEHT E'tperlnont HT402 i RcnanBra by inhjlaiicn to VinyllAsno Chlerl4-i (VDC) in nil* t WO, 100, 50, 23, 10 p?*, 1 hourr felly, (-5 fe.V-3 w-tekly. for 51 Kooks, Heetilt- ertor 121 weoka (oni of experiment). KCK nmr.USTT-1 I.FSTW3 0* SOWKT /.MIMICS {Swiss rplca 1C *:re old (Cro T, II,III,lV,V,Vl) o:id 9 wkfl old (C ro IV bio, VIT) nt ulr.rt ) Exav-lned AitnoiLS -riTK :iou rsopustio IESI0HS OP KIDffSV Rcgroor.lxs chanJ[0S Total Amyloidosis (a) ++ (0! Chronic nephritis Total A- <*> , ++ M Concentration Lervyth Sox Ho.at Ko, ft Ho.` 1 Ko, St Ho. ?. Ko. Jt Ko. * Ko. :> Ho. jt stort \ tv 2fi njvt. 52 ttorVr P* <1 30 29 53.3 0 - 05 07.3 12 42.6 13 46.4 16 64,3 30 30 100.0 1 3.3 2d 70.0 12 40.0 12 49.0 19 63.3 5 17.5 13 46..*. 9 >4.0 10 31.3 IV Me <5 pm V 10 ppci _______ Ko troitxonfc VI (Contn'o) 0) I rL.TJt31 5e treatment {Cc~*.r;Zsr 1 C* oral 0 CO 36 9C.7 1 1.7 O' 120 110 91.7 - 5? wool:: 9 O" enti J 1.70 112 9313 240 222 92.7 00- * 52 woote a' 9 0" and <J 30 23 76.7 O' - 30 73 03,3 0 CO 45 60.0 0 ; - O' 9 O' end 9 100 02 G2.0 100 09 69.0 2C0 171 05.5 00- 0- /^ O' 50 0 9"' ? 1 it; 61 90.0 0 ' - n3 r?.r l 3.1 M4 30.C | _li 1.4 |l _i 930 j 1 d9 67 .5 2d 70 70.9 43 77 66.7 12 155 59,6 53 21 91 0 11 41.4 39.1 .10.7 24.6 47.5 35 63 100 10 'i.1 -J2_ f ,1.9 ji.n 63 31.3 56.0 39 37.7.- 45.0 122 34.9 ~\r43.3 73.9 ' 19 76.0 4 16.0 15 CO.O 11 44.C do Cl. 3 13 J1.2 85 57.1 26 56.3 ' 59 71T9 22 "m'.T 37 ^ 45.1 57 69." 59 G(S# J 22 24.7 .17 41.6 61 66.S 116 69.0 44 25,7 74 11>3 118 69.0 44 51.3 29 '2 0* , * *- 1C To 52.6 43 35,3 * * 15 3 !f .7 | 23 13.3 49 59.2 11.4 ** 3T*T 15.5 7) 50.7 14 19 40 4 J 7 20 24 44 34 *< 5? f 1.1 JK 19.4 *7 36.2 17.0 *c 35.7 19.0 C2 |36.9 17.4 12.0 14.6 24.4 13 36.5 0 32.0 21 31.7 V U5.1 24.0 37 41.C 25.7 74 lf^-3 j'.r fiTh:.r r;.J| r * '* * 36.l| 21 14.5 11 {*} * 1 ailil ) narked. BUHTBT'nO'T `nrTTr.r t--;i i'fBJ ij. i 'im LV Ul . |. >vqns ' j V J . .1 I a j i'I ti 0 0 CMA 0 1 5 6 7 4 (e.) Alive titr.Lln after 1'J ne:re, when the fli-at txucour (a leukaemia) me oj.Mrv.:c . (;.) Tm> or r.or tu-'ourr of tho s inn cnj/or different typea (ftbrondenomna, cs .-cinema, nrconn, carcinosarcoma a) ma/ bo preaent in the Sana ani ral. l'*/ Ton rtncnlnnno are referred to tha corrected number. (d) Average up* ot the onaat of the first mcer'.rr tucour par a-iir.i"., de tected at tha periodic oontro\ or at eutopry, (e) Avercc* tlT.a from tha start of tha erporliter.t to tho do sc cilia Jet tha perintio oontrol or at ajtopay). | (f) Several anlsale with two or acre tunoure, j ____________ .` J BtreTBTT10! *qsT*I1B<I Ti0IS'eoT1B3 mm,, -purKl WI___ A SSOH i v-ifq^s $ D dp cV Rxperi-r'n; ftr>,'' 'pOv : by '.'..alaiicn '/Ir.yli:, Chloride (vuc) m *i; at liu, K`>, f.', 25, ic pr*, 4 c?-.i " daily. 4-5 d,,yi ..cct'.i fcr ;a v. Vcsulr S :i .r.' ' j'.Os-Vt (*i;d af d. ~ ?xpri:it). sis t*j. r - .it :o:s ,, r th; :c CQTir.-i r.-o';--.: Gzourz KO. sex * 9nitfn JMo. Distribution o:' i.; j teiypei ctt:... C'jr.s A.J ignant Ii : -liucion of his to tyros ! ' a" 1 lyr.bal gland adansa* 5 3 derratof ibrcras 1 adranal pfccaeP.i'unocy tor-a i suh.-utaaeous sorcor.* 2 1 ivi toneal T.asoiP.elics* i 1 2ymbal 31*04 afeitcn* 91 1 istestrnal adenocarcinoma i i rvarv fibrosarcoma 3 1 tarcir.ana of the uterus i iUtrnodoainal sarcoma i Il'Tbnod* ar.giosarcor.a 3 dermacgfibrcians cf 6 1 forestoaach p* pi Hoc* 1 Leyilig cell* luscur 1 lymphnod* ang; era 1 subcutaneous sarcon* 2 1 Whereas adeiocarciaom* ii 9 2 1 adrenal eortic.il adenosa 1 urinary bladder rapines;* 2 1 i.i tribdanir.al sarcoma i tt-'clacra of the uterus I j r I. subcutaneous lipoma 1 intrabdoamal llpora 1 skin carcinoma | incrabdcainal liposarcona ' 0* 5 1 forestanach.pauiiioe* . 2 1 spleen angicr.r. .i 1 urinary bladder pupillcsA in 1 92 1 2yt.l>al gland ader.ca* 1 adrenal phaechrarecytar:* 2 1 o, --. '/ arrhenablastoma 1 cticiiirss of 'he uterus 2 dermacofibrosus 4 1 subcutaneous i i>ar,a 1 Leydig cells t-inter IV 9 1 1 fortstemach ; :c.v.a 0 0 <t 0 V 1 subeucaneous 11;otts 9 1 2 ! :"uS'.'".lancou5 'arcana : i-'^or.ai pheoahroKoblas tear 3 * ir . raedeoinal s-irtojia 1 lei ir-i/osarco.-.*. n? the utiru. 1 darmacofitm-:'.:, P ;'HbCil*-i*suS Sin'mall * 1 subcutaneous ip''cl 1 s.:i v-'ory glare C-ireinosa J 1 subcutaneous fibresegieci. 3 l . isl*noc*rcinor:a 1 forcstonach 11 era i L-.. .'.'bicarnal '.arsotu 1 adrenal con.--- iiifti----i 1 r-aal. phe oeii roir.cbtas toil* 1 r.ii rj outer rrsei'ieiiov.a i ) 1 CiMa t ;:eu i'i isoisa VI 1 Zyiital 0":. 1 . ;I eorti-A l earcirr.or 9 3 1 caTt' ' I?*!!?.*** A f. , ;m35 of ut-.-'"ii Subject t- ' ' Fdss v. C~ 14th Jjii - ' ; Calcasieu Parish, - 3i i J7 Louisiana CMA 015675 \ wtccit r' Experiment DT401 i Exposure by Inhalation to VJnytldcnn Chloride (VIK!) In air at >50, 100, 50, 25, 10 ppn., 4 hours dally, 1-5 dcyo aookly, for 52 wools. It caul t* aTtor 1J7 weoks (end of experiment), . .. . MO. I I: T*T IV / THAT IOM 150 ppm i 100 ppm J 50 p?n 2> Pf" DISTBITIUTIOH Of fHF. EIFFEBE3T TT?E3 OF HAMTAnr TUIMUBS JU1* 4L3 { Spraj\:e-Davrt cy rets. 1C wnnki old at stort) Corrected Sox Ho.at etnrt iHinter u) To tal Ho. Avonrfl lit) p 01 latency tuuourc/ i. tin# (0) (week#) bsoring (j) nnInals O' Cford (? fO 60 TJO JO 60 3 ij.j 97 + K 1.0 60 u 73.3 821 J 1.5 ,s ^ IPO *i.l 02 J 1.4 JO > 16.7 104 i: 9 1.0 n.VWJIT TO HOURS (bj Histologically examined !!lstotype Total Ifo. Jt (> 8 100.0 1J 97.7 51 98.1 5 100.0 fibrondenoxas Caretnoraa Average Avorugo latency latency Ho. Jf ' tine Do. C time <r> {treat*} (f) (nooks) Ti) (<1) 6 7> *0 1091 8 1 12.5 26 33 88.4 83+ J 9 20.9 78* 8 E6.J 76+ J 10 19.6 73* 8 .. . 5 1C0.0 104 + 9 0 - Saveokns Average Intoney Ho. Jt ' tin* (f) (necks) (61 0- - 0- 0- - 0- - ----- - ---i __________Ii J I * Carddoji. .-e< r.-.ifl Ai (.r-j. -j *1 1 Pitney llo; t! ^.e (fj i -. \ * '.1 76 0 -11 - 1 7-S c- q JO JO y/*3s 8J.3 B2t 4 1.7 2J 92.0 21 91.J 8J 5 3 13.0 102 t 10 0 - O' nnt* 0 oO _C0 50.0 05* d 1.6 28 9 J. J 26 92.8 871 4 3 10.7 102 110 0 - '0 JO " J.I 10C+ ; 1.0 7 100.0 7 100.0 105 + 5 0 - - 0- - 0- - c- r__. 1 '~0~ - ? cf a-.d 9 O' JO CO JO a [ ^ 76.7 79; 1 60 fltj ^0.0 061 1 u 28 14.) 10JJ10 1.9 1.7 t.0 22 95.6 21 95.4 821 4 t 4.5 C8 29 96.7 28 96.3 801 4 T 3.4 68 4 100.0 4 100.6 10JS 10 O - - 000- - c- c- 0- -; *t 1 0 (/ and 0 30 60 JO 50 70.0 86+1 4J.* E81 4 1.6 1.5 20 95.2 20 100.0 871 4 4 20.0 82 * 10 0 24 96.0 24 100.0 90 4 .4 16*7 62 4 10 0 - - 0 '* - c- -1 j V Vi Vlu l 0* 10 P|IT* 0 , O' niJ 9 i;o tresj: tf meat fCon- n t roii) Cf and 0 L__ JO JOCO TOO 100 ?00 SCO 25 v j 10.) eu2j JO r 50 93.J 8 Ji 1 52,5 81+ 4 07 | Ti >2.6 1151 6 99 i 6' 106/ f7' 51J ^ 61.6 07* 2 JO .7 $1* 3 .. 1.0 1.6 1.5 1.0 1.5 ......... J 100.0 J 100.0 81 23 0 - - 24 85.7 24 100.0 85 4 5 20.0 90 14 27 C7.) 27 100.0 331 4 5 '0.5 90 * 14 1 1 100.6 11 100.0 115 6 0 - - 5G 91.8 44 78.6 88* J 16 20.6 93* 5 67 9J.0 55 02.1 93 J 16 2J.9 95* 5 0 0 0 0 0 0 - - 0- c- -j - 0 - ~~1 - 6 -. - 0- - - 0- - (n) Alive enlmnls after 10 wreka, when the flret tumour (a leucaemia) nao observed. (d) Averse* *S* * the oneet of the first manner/ tumour por or.lr.nl, detected t the perlodie control or nt autopsy. (nj Two cr more tumourn of the name and/or different types (fibroadenomas, () The percentages nr* referred to total number of animals bearing mammary corelno-sae, e.irconaa, carcinosarcomas) may be present In the same animal. tumours. tel Th* oircrti,. icl .he prrcr.tnreo ., referred to the oorreot.d number. (f) The poreentages ars'rsferred ta total number of ar.lnalabearing namery tu noUre, hlstologloally examined. ...... s. 1.. .......--_................... ............ . !. -------- . fS S) If) U t Q butt r,rnci7 l.'i *' T ' T r H't ! r: TT; > j-pljj L?r 1 < r inn ut n ssojf 1 i,:'>'qag 8^9510 VW3 ijlblb 12 i o' Sotcc, Experiumr.t BT401 t Exposure by inhalation to Vinylidere Chloride (VDC) in sir at 150, 100, 50, 25. 10 ppm. 4 hours dally, 4-5 days weekly, for 52 weeks. ffgsults after 1J7 weeks (end of the experiment). BTCLOGICAL PABMfSTTSS RSLATCD TO HAHXAEY TUMOURS a * r: 15 {3pra^ce-tU rlcy rats, realms, 16 wteks old tt stirt) t Zcz *ec^ v-ir:Uc `"ale A t ei rs. v; th v! "kc : r > r.t.irt (*', X^~ nu; n I ,, > ;;1 ' v.j | be c - AVERAGE ACE AT THE ONSET OK THE T01?OUH^ Total Fibroa dmi>o- nK) Carci noma* 3*rc. Carci net nosar comas *; : 3 3>r 3 7 - 1 - : KCAtf VKICIIt (g) \ eiontns X> souths V/i i r e 1 s Anlraln .5.1 trail Animals wi th without lth rlthcut tumours tumours turrotifs tunours 34B 967 394 9 go AVERAGE AGC at ns.\rr n ,,n* :'*v.i::a'y a dt::t Gvc*r; w.% n:y.\M *;; :i .? v;r-. rci A` V :vjc 13 r.c:.13-J 1 ir,inlz Ari-alt. A:ir.j.!;. `.'.lijit riifl .-..'.r.i.t with ;Ithou: tlrh :! the Jt rj-y;i! j v:t i tunpurs tumours tu"!our'.i tumours ! " I -l .* t * . 3S<? .f 3S5 f iff :;i s: - ? s - 1 ` i |,or; :c 3C |:.5 ' j - j 5 U.7 B!l i tj! J icj iio - 9*3 7C3 337 309 339 19* 1C9 lA 9016 t "FP........ s . i* ' ; K , in jo rv t :.~j j 53 tr.ii) i ~T .... . . , t . ; ; 1 70.0 ' 9 30.0 Ktt -5 B7i 4 *a;io v,l iJ 0.7 jjt *' SOT 1 *1 G! 51.6 33 D>.4 37i f Ml ] 95l 3 - - - - :'7S 2C0 1>1 :( 3G1 33-. i 179 975 330 336 379 1(0 105 t -1 j Tl;i: 1 763 975 11* 335 339 i::7 105 Id | sc:2t 1 1j ' 1T :: | ' 1 J9(8 . 974 39! 115 371 3(4 107 i 9 7; ; * 13.1 ; ?.s i 't '.live arural a after i0 reoke, when the firot tuaour (a leukaemia) was observed. 1 o VT- )' W< : UJ J ` 'Jin ''1 ' 1J ft "ssoy 1 ';1 _V>o{*qng fi V I ,!, 1.1,: a J ,=T jj rj ') lAtiw: t j CMA 015679 (b) T*-o or T.cro tuTOuro of th* non* rind/or different types (flbroadono. ra, carol noons, earooTjie, carolnoonrconne) may be present in th* tan# nnlrial. (c) Th* porc*nt#c*s nr0 referred to th# oerr*ot#d number (#) Avernro tin# fro* th# start of the experiment to th# detection (at the poriodio oontrol or at autopsy). (f) Several animal* with two or more tumour#* T Q 1 zm- r- . U(. .1.., ' 'it/r A ^;;og ' i l A-ji qns IV I J. t'i.sfii j V 3 G ^ *o SABLE 1 4 Exparir.er. t 3TAQ3 : Z<: o V-J r ? by ingestion (stsmach tube) `3 Vinylidcne Chloride (/DC) in Oliva oil at go, ig, j mg/tg tody vmg'-.t, once daily, 4-5 days veckly, for 52 vecks. kmvi 1 :s after 147 vecns (end of me ox penmen c). Miwm Dmimrnrsv-fiori ' ck1 " oi1r1?o?.z"'r": "r? ?~i" o'"r .1 (rscsui .-A.ivn sc--ys iiCi ooiut TMit-) "ru11fousa" GROUPS XO. see NO * ANIMALS 3ZAkINt3 QtHEP rLKC'JKS Benign Distribution of histotypes Malignant "o. Distribution of histotypes 2 1 skin acanthoma 1 dermatofibroma 1 i 3 3 forestoaach papillomas 2 1 subcutaneous sarcoma 1 intestinal adenocarcinoma 1 2 1 ovary carcinoma 1 carcinoma of the uterus o' 3 darma toflbrcmas XI 1 hepatoma 0* i 1 adrenal cortical adenomas 4 4 subcutaneous sarcomas 1 intrabdominal sarcoma 3 1 adenocarcinoma of the uterus 1 pericytosarcoma of a lymphnode o' 2 2 dermatofibromas III 1 adrenal cortical adenoma K 0* 1 1 1 thymus parleytosarcoma 2 subcutanaous sarcomas 4 1 adrenal cortical carcinoma 1 adenocarcinoma of the uterus 1 skin sebaceous adenoma 0* . 7 2 dermatofibromas 1 subcutaneous lipoma 1 2 forestomach papillomas 1 Leydig cells tumour IV 1 1 Zynbal gland acanthoma 1 intrabdominal lipoma 1 hepatoma i 40> t 1 forestorach papilloma 1 adrenal cortical adenoma 1 adrenal phcoehromocytoma 1 adrenal gland ganglioneuroma 1 leiomyosarcoma of the utarus 3 subcutaneous sarcomas 3 1 liver angiosarcoma i adrenal pheochromcblastcaa 1 ovary carcinoma 2 adenocarcinomas of the uterus 3 Toca: * 5 7 5 3 5 TO 13 J 4), TPKTITT AL -d! CMA 015680 f ft 1^1 " S 'd ? ' TAJ3LE 15 ;erinent BT403 Exposure by ingestion (stomach tube) to Vir.ylidene Chloride (VDC) in olive oil at 20. 10. 5 mo/Kg body weight, once daily, 4-5 days weekly, for 52 weeks. Results after 147 weeks (end of the experiment). INCIDENCE OF PITUITARY ADENOMATOUS GROWTH GROUPS NO. I r n CONCENTRATION 20 mg/lCg 10 v.g/Kg ANIMALS (Sprague-Dawley rats, 9 weeks old at start) Corrected Sex No. at start number (a) CT* 50 An 9 0T and Q 50 100 45 97 V" 50 4 9 (f and IJ 50 100 50 93 PITUITARY ADENO * MATOUS GROWTH No. % 2 4.1 5 10.4 7 7.2 3 | 6.2 0 |3 j 3.1 0* 50 49 1 2.0 iii 5 mg/.tg 9 50 30 O' and 0f 100 99 7 14.0 3 8.1 (f 100 100 IV Olive oil (Controls) 9 ICO 99 rf* ar.d (j> 200 199 4 4.0 9 9.1 13 6.5 (a) Alive animale after 14 woelcs, when the firot tumour (a mammary carcinoma) was observed. .. . .......... ........... . . ... O t A H J i h* IO CMA 0 1 S A 8 2 (n) ^tlvo .* Tt?r Id neot:, whn the first tunnur' (& Bamarjr eart n*T,i) nil obrvrvjd, (tj Tvt3 or Mil Ui%>ur of th* otjto and/or different tjrpeo (f4 broadenoaas, tr.ruinonaot nr.rcoMit, cnrcinoaareonas] nay be pront in thi eun tnl* nrtl, fc^ Tha poreantcgee nra referred to tha corrected. ntrebor. (i!) A;#rsje fi(,9 at tha on**: of tha first irnwrjr tunour par sulcal , detected at tha periodic control or at autopsy. T (a) Tho pcrctntnj** tr nfm4 i total nuefoor of anirslo bttrrlr.j \3u2rA?;.' tumours* (f) The peroartagsa nra referred to total ittwbor of anlcsle biitritf tnaar; tvaours, hi otologic ally ixulmd. ({) 1 f lbrooarcoro. (h) 1 flbrosaroonaj 1 perioytooaroeoa. v O Tt-t't <> irmT . -TM' ___ T fv SBOH v ." *C<?' 'f 7 totccxt r~\ Ir1 TABLR 1 7 E*fr|jfnt BMC! : Cxat tsre by ingestion (stomach tube) to VinyliJene Chloride (VDC) in olive oil at 20, 10, 5 "?/!Tg body eelght, once daily, <-5 days veeVly, for 5! seelt*. 7tult; after 147 veeVt (er.d of the expert nett). BI31P3ICU Fh3.4i1ETSB3 BElATriD TO KAKH.tiir TO^OMBS n GOO m < U !' 1i i; i I ,1 t ! 1 9 i ' I- r*;=.rt I 1 1 _, l :>' ; C.;rr*ct U) i i V lit X r" r; " Ar. lirAl ^ jit hout m O'lVe ToU! "ti. *rt jl. .'..I.* rtV >Tj .-r ;ii: vuxsius Fihro H,.t 1 >ircl- Circi- far-co rtGKJt5 p.ar ros it rtlO.-fil 4 ROi trlS 6 months \niral i Allmtli Anirjlt Antval* * 1 r ii without *i tii vi thvjt tlKXrUl-g tuueurit rmt)Uk> tunour1; ` '2 wonth9 **11* si', Ar 1 tfi X + wi th ji l*K!Ut .u.runnti tu.jur". S''V ; i . . ArlTAlf e'jliTJlI* 1 1 til * '.i et-.-ur-: i *,v- Is - ' ! ;_3ir n * '] -7.9 }r* 53.7 71 i 6 73 i 12 - - 177! 23 33< 312 303 375 J4.| 951 5 75 1 5 * J i l_ ri-t* | 50 ,0 5'i 'i3 .0 if 57.0 6 71 5 sj: j 7# i 7 - - 26] 251 3M 302 363 346 111 t 5 721 6 ' - i i r ! i 7> 5*.0 AA.O 90 i e 38 1 4 ssi io 17* - 2J* 257 301 296 33* 341 110 L 4 59 i 5 5 1/.5 ** : i i i 1 "* t j 'CC t^o J_ _ _ 57 57-6 <3 4: ,4 Si- J c;i 3 SI -6 8 Til 6 59 251 251 314 314 557 343 1041 3 SOI 4 5 a.t i.i. | ilin inlanl! efter 1* nteho, irhtn the first tumour (e --n-y serolnme) neo obeervsd --TP' I m;' i 'l.t . ,1 )' ! i ........ 'T^O i ! 11 Hi__ iee^ O'. !! ' Ut J = '[-u0 ! i r' M r,'l TV IJ.NEai.il NOD lAHI-t IO Experiment BT404 I Bxnoanro by Ingestion (etomnch tuba) to Vinyl!done Chloride (VDC) In ullra oil at 0.5 rnff/Va body nalght, onco dally, <-5 deye noakly, for 52 nooke. He'.iultB after 1J6 (end of experiment). DISTRIBUTION OF THE DlffEHEH? TYPES OF TUK3URS , "PCJPIJ 1 CCMCE.1- \ l ->0< JlltlW 1 A>'ir.1At5 (Sprague-itetflcy nt9, 10 wcekJ old ftt.s^art). Sex :fo .nt rtart Corrrototi nunbsr (a) I '5 ^; J $ 1 Uf (*;>n (j> -i ii Olive oil (Controls) Jo'* ftnd ^ 50 . iff IOC 97 Ssf "t T 77 77 *59 158 . T*itr.l , 255 *55 (fe-rjeary tumours (b) Avon To- < latency tut Ho. (c) tins '"UT' * 4.1 K>5.;y,1 t. /. 45.3 C<i 3 4 24.7 8C1 4 e *.5 ! 75113 35 44.2 03 3 36 22.8 ej+ j - AHIHAI5 WITH TU'TOUnS 2ynb.il Gl=-id ccrclnorma Average Totil no. * (o) latency time (^yo) Totut Ko. 00- -* -1 leukaemias A/ora,;.' ~1 Other tiknouro 1 ~l 1 Total (f: ` >, t; a- r- S! Itteniy Total 3P"lfjr. ,:ail.r <i v..-j (c) time Ho. ho. TJo. (c) i savor../ no'j.v t t'lr:. -l is) r rt.V:: i`:t! 4.1 or-it n2 0 -- i f e. !' 2.1 in r4 0 j ; ... t ' 1.3 i 0- - 5 3.1 100 7 6 0 Sa 70.5 j1.2 0 _ _ 4 4*<> HP-1- 1 1 1 1.3 90 2 2.6 8220 1 0.6 50 6 3.8 66*14 ------- 4 5 1 3 38 ;9*o. 3 1.1 -- 2 3 45 |75.T 1.1 i"` ' i {a) AIItj nr.innln nftcr 70 reek*, when the flrot tumour (a l.iukaenla) iraa obeonrad. ->0 or nor* turcurr of tho cai-.o and/or dlfferast typas ({.'.broad an ana a, carcinoma, aarooma, oarolnoaaraoaaa) nay bo praaant In tha air aiAtal. ( = ' The percentages ara referred to the corrected nmbar, <!) .'.verogo age at toe oneit of the first naxnnry tumour par anlaal, dataoted at tho periodic control or at autopay. () nreroga tin# from the start of the experiment to tha dataotlon (at tho periodic control or at autopey). tr) 3<i--eri anlnals with two or aora turnoura. ' I to>lHirool o < U i f 1 >- 5 prrr' [ r -i.u'j `UfiTOTJ TieiSVOIBO ,1 * - i T-rmrinp qq^ a psog J' [ '^g ri V J J S 8 9 S T 0 VW3 TADLE 1.9 Experiment 3T404 : exposure by ingestion (stomach tube) to Vinylidene Chloride (VDC) in olive oil at 0.5 mg/kg body veirjVit, once daily, 4-5 days weekly, for 52 weeks, al ter 135 weeks (endj of the experiment/. i?TsT.;i5;irtor or ths Dirrs.tsaT types of miscellaneous ("ot'cr") tumours : CrOUPS NO. se:^ 1 I II 9 &' 0 t ANIMALS PEAS'tNO OV.ILE TUK:>'.;R3 ------------------ ------1------------------------ .--- Ber.ign Malignant t!c. Mo. 0 2 1 skin carcinoma . 1 subcutaneous sarcoma 1 subcu tanaous sarcoma 0 4 1 liver angiosarcoma 2 intrabdominal sarcomas I 1 adrenal cortical adenoma 0 Total 4 \ 1 1 forcstomach papilloma 1 1 kidney adenocarcinoma 3 1 adenocarcinona of the uterus -1 1 fibrosarcoma of the uterus t;uu i*1 : 1 ' rtf' ? T1VI o I- !' I . i ( 'M'L____ im-oe ` ; "1 '>'Xc-'H ur aet.,io f' 1 i ' *1 !nS ji n ft a i I no d TABLE 20 Expert irent 1JT-104: HxriosuJjl y ingestion (stomach tube} to Vinyl ider.e Chloride (VDC) in olive- oil at C.5 rce/Xij body weight, once daily, 4-5 days weekly, for 52 weeks. Results after 136 weeks (end of the experiment) TECIi-EWC-i. O" PITWITATiY ADd,7C:'AVJl'.;;_ GRQ'.;IP * GROUPS WO. cc?xsi:tratio?i .ViZ.Y. (Cpi\:ijuc-i\v/l sy rat, i0 weeks old at start) Sex Wo . 2 t start Corrected r.arbor (a)... ,, 0* 50 49 oin i .i c, j i____ 1 1I I o.5 rcgAg Aft tf ar.d Q 100 97 i 1 11 Olive oil (Controls) o* 9 rf and ^ .82 77 159 81 77 159 1 :c j KA'IOUS GF:0y 1H | 1 11 h'o. | % o 11 __ _ c 12.5 6 6.1 0- 2 2.6 2 1.3 (a) Alive animals after 20 weeks, when the first tumour (a leukaemia) vias observed. >C0O o iHn <ro - @ , r ,r noisKoi^O -,rr UU'T__ *" LSR-fid. ' ` ut uey.io "MJ- '1 Rf?oa t .tig "'fviARSUl; l .-1 He.- 'v TABLE 21 roso >ti0l o <ru *Mve tnm,il9 fifter 20 vtecSo, when the fir at turjour (i loub&wxla) was observed. >' cr r:crt iuv.o;ir< of the znr.o tni/ov different types {flbroadeanae, oarelAOfaast ureoiut,oarclnoenrcomas) uy bo present In the cstm onLail. (<) r.ii pro(Rt'ijD ere referred to the oorreotod .number, * Av^rnpe net at the onset of the first xAfonarjr tumour per anirtel, detected at the perlodlo control or at eutoper* <oi Tito ptreentd^it are referred to total number of animale ^innj naieniry turnours. (f) aThe perccntcges are referred to total r.uabor of animals boaring mammary twourt,bletologioally examined. * a \ i n ntn svo 1 . 'HH___ A ssoa AEUi" nr, ut ;1' i ' i '-tf-ins 1aMO3 TABLt 22 I 03 00 >0 tri43 o X. o - -!1v> ir.mli after 20 wo*a, when tha flrat tuaour (a louVatala) m obaarrel. t 9 " 'T-'r T T''''TfTT:o^B3 Q L,-'[ ' Utft A SKoy J. . ' 1 i v, "H'qnjj fl VI lL W ,>J 0 / ,ti 4(i 3 o yV .: 23 m&s!i,/?.7fI'i\ieporirien: ZT'.O'i : r~;*-. rif*.~" i*/ ; st . *.G !.'*** *, <M -t'k * *..> Vvt'./T -jc n-iar^ 'inl/# n (e*v* >f 'ji.'.oriilc (v;c) 4 5 lays r.eel;V, **.^r t). in iir t fir 5? ..c SQtCCNi .Tv- c/rjtvvjKS TKi-A o* K ft K<* 5H *fVh o= o `4OUo I PP* days II PP3 days III 50 1 ppra TrtCk IV 25 52 ppm weeks IV hie 25 PPO 52 neoke V :o 52 pjxw weeks VI VII Uo treat ntnt(Con trol*) uo tr*at went(Con trols) Total A. ..t wi . , ,* t <` f t 4 mil 1;. r' *'. ?lw (Ors 1, TI, v,/i) a.-: '? ..C.'ra old (5:-t I .* v* *. n:> tt " .-M, -** S*x utirt (fir.' A.'-1," tL* " i:k iur ?LMS ft u'\ *,* ^U;f* ' C . T'21. , , ; Jt I'ir-nrj- V^Y.OUi y *o* Vi 'r r* o^ X t n a*-" . -* n 2 * A - >t X u C C?2 -3t * (i -- S I * .5 w oo % 0- uO 'l 0 - $ 0- 9 0* er.d 9 60 2C 2rd 0 2-3 - - 53 1 l - 9 59 1 1.7 0* 0 - 9 Wn0" - O' el <1 60 23 0 - - a' M ^7 1 "Ts" 21 23 3 10.7 49 3 3.1 27 0 - 9 H -- O' and 0 CO 3) M 3-2 :* 23 53 TM-y :o 21 W' t ;.3 7' I 5 29 2 7.1 2 3-6 0 9 O' er.d 9 30 so )0"" 2<> n A 3i "rT' O4 ' 2?. 5 - 30 7> t 5 59 752~ Vi"" X" "T : 13.3. 318 ,Q-8 9 uo 112 1 0.9 77 118 12_ ; 10.2 O' and 9 or" 9 O' and f; 9- o* iin-i : ~~ ?-o 30 33 10 MO tec 1 9 0* and 0 C-t 210 26 25* _ 0 26 0 51 0 53 1 <r* "" T> Q 0 ir*. 0 155 0 12. , 7Si 2 235 - 30 _ 30 -- ' ' - * 60 92 97 r: * 159 80 88 T\ ! ... . *,n i 13 U 5.5 0 >l:r 6 ,20.0 s ioTc> 1 >rl 2 , 2-1 3 T.6 0 1 1.1 1 0.6 (a) AJire animals rh;n On firn*. V.-..- ' .tbs or.,:.rad . r J\lty i:"iunnnr-itiois.'i : 55 ws*l:s; unary tusour : 77 w*ska; fi1". mry .id e **::: i : Id -..>.; i sv'.-**. na : 12 vree>.j.` The pareanta-ien r.m rtferril l i-:w curruf.Jii- nti-r - . ThO pcrrentflija:' of (.oral rn-.vni t.i : !> -ini'..-In .`-.jli -lira at the !2lh >-J*k , when the firnt tuB'",r - wan h.;i"vad. (h) Avara-jr Use f.--:.- Un star trol oi* fct auto. ijl. rap'sr..tc 11 ! .-:.. c-.n (r.t the p/"iniia eer.- rmnPTDEHTlAL. ----------- Protective urler in Subject to 'To. 50 --133^ Boss, 7. ^i-L-V" . j,laT_H-,i-t Court- 141h 1 ,,J ^ "Y^- - 3iana. . Cal' asr - P-` i-"- CM* 015689 0 o 23 :;?<C2 : ^L'lTS ^ *q I'l.iylideTo Chloride (VT>C) n K,- .. 2C^ >> '* * 23. ' - *;-- d.in-, 4-5 dv, *a.kl/t iUV ** 30LCSHA | 7 .fj/ * .V : . :t ri?:s ci- rnv^s sO o u I A.>:r Vulnon'u/ adeaena: ( ; ) <3 O2UilOa*. ^t*tve.l wt a3s Cl :n --0 V. O1L>CV*<J. W!*zNv JV-_CuC S 0- - 46 1 2.2 57 53 1 1.9 57 a | IOCld-l C4 s - ` 1 ' S = j 9i 57 l 66 1 d 3| j| r.'ci;?.s ~ JT ' #>;; ^?c';3- w:v*9r - 1- 5.3 | 3, 416 4.5 j 3; 016 hWC,| 3| Others * *OI-f* --A<&Hd*. Sa Cotil (4) %O (V ^e O 3 0 Cc 0* i .a =- ,, ^ "<i UC c a 0 0 0- 0 2 2 0 6 10.5 1 2 2 0 6 9.1 1 10 2 11.1 621 7 23 | 2 | .7 j 23 1 7 0 0 0 4 17.4 1 II 26 2 7.7 53 i 2 30 i 2 | 5.7(2-316 3 \ 2 8 21 .7 1 44 4 9.1 531 4 53 i ; 7.5 27 . * 3 1 2 12 22.6 1 i 'i: 26 1 3.3 62 29 | 0 ~ - 0 0 0 2 6.9 1 ! i III 27 3 11.1 80 i 2 29 j ; | 10.3 ( 110 3 ! 2 1 10 34.5 1 53 4 7.5 751 7 53 i 3 } 5.2 j t 4 110 3 | 2 1 12 20.7 1 i1 28 7 25.0 7315 30 j 2 6.7 j -.9 ! 20 I G 1 1 1 36.7 1 IV __29 7 24.1 C5 i i 30 | i 3.3 | 72 9| 7 2 17 56.7 1 * 57 14 24.6 sc; 4 60 j 3 5.0 ! 5* S14 .10 | 7 3 23 46.7 1 l 113 16 14.2 7'' I 3 120 j 3 "i. 5 [7*TT 5- 2 "43" 35.3 1 i ft IV 8ia 118 231 11 9.3 73! 5 119 j :J ! 3.4 1 52 Z 5 "1 7 27 11.7 77! 3 239 j 1; 5.4 4 1 & | 12 4 40 33.5 6 83 34.7 1 1 28 11 39. J 711 5 30 ! 1 "").3|t'.' 0 * 1' ( 35.7 1 \ V 30 3 16.0 i8i4 30 | 6 ii-j ''~j i 7 6 3 3 15 50.0 1 58 14 24.1 70; * SO . 7 | 11.7; Tii i 7 1 J 4 25 43-3 1 i\VI 80 92 3 3.7 c - 7 99 i j 4 4.3 561 V3 i 7 - i- * 7.; ! 41 r 5 i | 1 1 7 7.1 1 15 >5.5 1 1 * 172 7 4.1 ijL' - 7 1S7 | 7 3-5 .!= 3 | 3 2 22 11.2 1 vn total 73 n 3 86 2 159 5 _47l" 5v:; 2.3 7f :i2 3.1 C'-i i 87 ; - 39 j 7 17o j t. tlHHVi * * 9 1 j 3.-, MI 5 7.: ; 4 8 *-"i :i - 7 k, -- i '1 3 4j 3 5 n> 1 7 .0 1 12 13.5 2 19 10.5 1 1 1 () Saaa of which with calm-.:- atypin.*. (d) Sovoral amelia wita *.t"i *cr<i 1 :"v r'VJP'IDENTI AL_ Subject " ^active Or'ier in -.337 Boss v ~ ' 14th Ic -:: ' ^ . Calcasieu P'-ri--. - pama^vn W CMrt 015690 69ST0 VWD L M < 7 'inr.Trj Q ' i` 'im ITS [ ( i ' f';i ` .11 ` o, u.r) a ssoh ^-'0 i.Mqoa-icMH n( jaefqng IVIiMSQJ 1U00 TABLE 25 Experiment BT402 : Exposure by inhalation to Vinylidene Chloride {VDO) in air at 200, loo, 50, 25, 10 ppn, 4 hours daily, 4-5 weekly, for 52 weeks. Results after 121 weeks {end of tho eT.psrir.VRt). DISTRIBUTION OP THE DIFFERENT TYPES OP SUJfi'S/.BY TCKOURS (eouond part) MO* >1101 <r. o (a) Alive animals after 27 v.'eeks, when the first tumour (a Kiaaraary carcinoma) was observed. (b) Two or nore turncars of the sans and/or different typo (fibroadenomas, carcinomas, sarcomas a.id careinosarcomas) may bo present in tho sarro animal, (c) The percentages arc referred to the corrected number. (d) Average ago sc the onset of the first mammary tumour per animal detected at tho periodic control or at autopsy. {e) Tho percantajue are referred to total number of animals bearing mammary tumours. (f) The perconteson are referred to total number of animals bearing mammary tumours, hi3tologicsily examined. * ncr* ! ' . * * s * . E-< (P! W Pi M Pu fe o 'I s:;! ^ " o +0*,-> I, , ^ 3,^,J 1 IIs'5 w| O CMA 0 1 5 6 9 4 TABLE 26 Experiment BT405 : Exposure by inhalation to Vinylidens Chloride (VDC), in air at 25 ppm, 4 hourB daily, 4-5 days weekly, for 52 weeks* Results aftor 157 weeks (end of the experiment). cncurs CONCENTRATION HO. ANIMALS (Chinose hamsters, 28 weeks old at start) Sex . No. at start Corrected number (a) ANIMALS WITH TUMOURS Carcinomas of the uterus Others tumours Average No* * latency time No. (b) (weeks)(o) No. Total Mean No. of different tu- * mours/tumours bearing ani (b) mals I 25 ppm d 9 (3 and 30 30 60 17 22 39 0 0- 6 27*3 108 + 8 2 (d) 7 3i#a 2 7 17.9 0 1.1 1.1 6 18 No treatment 11 (Controls) 9 17 6 and 5 35 16 11 27 0 01f 6 54.5 95 + 12 1 (e) 7 63.6 1 7 25.9 0 1.0 1.0 Total 95 66 V. t a) Alive animalo after 63 weeks, when the firflt tumour (a carcinoma of uterus) was observed* b) The percentages are Inferred to the corrected number* K c) Autopsy finding (from start of experiment to death)* ) d) 1 skin carcinosarcoma; 1 ovary thecoma* c) 1 skin carcinoma. eucTetno'i 'i|ST.ie,j najaeoiej i ' i < ; ` " WV Afi't -t.'G *:i ' :! 'A ssod u; aepjo ->i loefqns 'zv i j, wTdTa i --f nod oo BASF Aktiengesellschaft Un.-K- IfWl iFi Tgigfofl .C62D $01 ^V^pT-iiT'ungj Tii -164911 baaf d iZentraie) Tolggrammg: BASF L*dwiqsnafeflr"girt BiAkvefbmdung. UiAdgSierMraitar* 67Q0 LudwjgsfiifSA. Gironorio 545 C7300 Mr. Professor Dr. Cesare Maltoni Direttore Istituto di Oncologia Viale Ercolani, 4/2 1-40133 Bologna / <=c J c.c P J cc /.< kO'>/z <fo*8 h't'Ls /$*>* - - t*4<0rt<0weAwAi c 9*trtf FLD/M he-mi (04311 OO 21391 Statistical evaluations of the VDC studies Sent, 26th, 197 Dear Professor Maltoni, as determined at our latest meeting we are now enclosing the statistical evaluation of the experiment 2 (BT 402) . This paper has been discussed with Dr. Paddle from ICI. The tables start with number 24 up to 39; additionally, the tables 30 up to 39 (survival curves) are enclosed on a film-strip. Your3 sincerely BASF Aktiengesellschaft copy to: Dr. Paddle,ICI South Lodge PO 3ox 3, Ful3haw Hall, Wilmslow Cheshire SK9 1QB Dr.T.D.Browne,ICI Castner Kellner Works, Runcorn Cheshire WA7 4JS Dr.J.H.Wilson,ICI Plastics Division,Welwyn Garden City,Hertfordshire Dr.A.Collard,SOLVAY & Cie.S.A.Administration General,Rue du Prince Albert 33, B-1050 Briissel Dr.J.Helfenstein,DOW Chemical (Europe), Comp.S,A. Bachtobelstr.3,CH-8810 Horgen Dr.L.Rintema,DOW Chemical (Europe),Comp.S.A. Bachtobelstr. 3,CH-r3810 Horgen WNT - J 560, Herrn Dr. H.-J. Klimisch CONFIDENTIAL FuTi'Jgct Boss v. r-c , 14th :' vfftrtd Bomlmd Timm 3*Wdr Vor*4e*rar*, Qrtbu+jht Stttatrf Re* Horst Pommor; nWc#i iHw*ortorto;t; CMA 015695 $R4ozodttrrqorAt; Awi4o7r00M L-0*r.snal#n. 4/OnI3QM1M tg Ht A?P ENDI X STATISTICAL ANALYSIS OF THE EXPERIMENT 2 (3T 402) Material and Methods The data available for statistical analysis are to be seen in table 23. That is, for each group, the number of - animals alive after 12, 27, 36 and 55 weeks - animals with mammary adenocarcinomas (=MA) animals with pulmonary adenocarcinomas (=PA) are known. In addition to this, the age (in weeks) at death and for MA the time of tumour occurrence were known for each animal in the groups IV, IV bis, V, VI and VII. These figures are available upon request. The conditions at the beginning of the experiment are listed in table 2. Using these data the following statistical evaluations were carried out. 1. The survival rates at the intervals given in table 23, and the mean values of life time were calculated to evaluate differences between treated and control groups. 2. Tumour incidences at the end of the experiment were calculated. Additionally tumour incidences in different time intervals of the experiment were calculated to see if there are significant effects which depend on survival rates. 3. Calculations were carried out to test if a dose response relationship can be found. * BASF AJ^^^gesellscha ft, ICI Ltd -V-` ' ' - c7^r in , ;o. 30-4837 -1 0 ourt : -uisiana CMA 015&9A 2 The comparison of mean was made with a rank test of Krauth (/!/) Incidences were examined with the "Exact Test by Fisher" (/2/) and the Logrank test (/3/) . For calculation concerning dose response relationship see /4/. All statistical tests were carried out with type 1 error a of 0.05 and 0.01. A significance for a = 0.05 is marked with *, a significance for a =0.01 is marked with x. Because of two control groups a C identifies the group (either VII or VI) with which the dose groups are compared. Analysis 1.1 Survival rates for 12, 27, 36 and 55 weeks after start of treatment The comparison of survival rates of dose groups to control groups VI and VII shows that only the groups I, II and III have significantly lower survival rates (table 24). Thisindicates a strong toxic effect of VDC at these doses. Therefore, the further evaluation does not include groups I, II and III. The survival rates of the remaining dose groups are generally higher in comparison with the control groups VI and VII. 1.2 Mean value of life time As expected from the results of the survival rate evaluations, the mean of life time of the animals in the treated groups (both mala and female) is greater than in the control groups (table 25). With one exception, significant differences are only to be seen between groups which began their exposure at different ages, for instance the mean value of groups IV and V is significantly greater than the mean of the control group .VII. Even when comparing the means of groups iv bis and IV (which have the same concentration but different ages at CC.: r. P.C 3 - \T - in CMA 01SA97 the beginning of the exposure), there is a significant difference. For these reasons, the groups were divided into two sets for further statistical evaluations: dose groups IV, v and control VI, dose groups IV bis and control group VII. The survival functions of different groups are plotted in tables 30 up to 39. These diagrams show that there exists an interval for each set where the graphs have the same shape. In particular these are the intervals: male female groups IV, V, VI groups IV BIS, VII 90 onwards 72 onwards 90 onwards 9 3 onwards The tables 26 and 27 include the statistical results of a comparison of the mean value of life time as well as the incidences of MA and PA separated for each interval. The intervals above reflect the same mean value of life time, but the earlier intervals show a lower mean (significant as a rule) for the control groups in comparison with the dose groups due to an increased incidence of early deaths 2.k Comparing tumour incidences, two tests were carried out: The Exact Fisher Test and the Logrank test. The first test compares the tumours frequencies at the end of the experiment; whereas the second test takes into account differences in the survival curves. 2.1 Tumour incidences at the end of the experiment The number of animals with any tumours in dose groups are significantly greater than in the corresponding untreated groups (table 28). EoSS ? 5 i ina CMA 015698 n. The tumour incidence for male animals with PA is significantly greater in all dose groups than in the corresponding control groups. In female animals there is only a significantly higher tumour incidence in the two 25 ppm groups. The tumour incidence for female animals with MA is significantly greater in all treated groups than in the corresponding control groups. 2.2 Tumour incidences in different intervals of the experiment At each interval the incidences of MA or PA respectively in dose groups are greater than the corresponding incidences in control groups, at some intervals significantly greater (tables 26,27). 2.3 Relationship between tumour incidence and survival times Fewer of the differences between dosed and control groups are significant when the incidences of tumours are compared with the Logrank test (tables 28, 29). However the continued existence of several significant differences shows that the increases in tumour incidences in dosed animals cannot be wholly attributed to their increased life time. 3. Evaluation of dose response relationship k The PA rate of male group V (10 ppm) is higher than both 25 pom groups IV and IV bis, in fact, in comparison to group IV bis the PA rate of male group V is significantly greater with a 0.01. For that reason, a reliable dose response function cannot be established. A similar result holds for female animals concerning the MA incidences. For female PA incidences a dose response relation (probit model) could be established for the groups IV, V and VI but there is the following limitation that at zero dose C'' ' - ; ~ Tin j - 4337 TUft - .,,3iana CMA 015699 there is an effect which must be considered (key-word: Abott's formula; see /4/). That means three parameters have to be fitted to three points so that the validity of the relationship cannot be tested. Summary There are significantly higher pulmonary tumour incidences in male and female mice in both of the 25 ppm dose groups. At 10 ppm only in male mice a significantly higher pulmonary tumour rate can be found with no corresponding significance in females. There are significantly higher mammary tumour incidences in female mice exposed to 10 or 25 ppm VDC. As .the dosed animals survive, on the whole, longer than the control animals, these conclusions based on crude total percentages could give a misleading impression of the effect of exposure. The use of techniques which allow for tumour incidence being age-related showed that the effects of exposure are exaggerated by the crude percentages, but that significant effects still exist between dosed and control animals. In general no dose response relationship can be established for either the increased mammary or pulmonary tumour incidences. The lack of a clear cut effect of VDC either in different intervals of exposure or at the completion of exposure underlines the lack of a dose response relationship.. 'r in , j. -J- 4337 i 1 ` - ` "virt Cal-'` s ' "'i t luisiina 25.9.78 CMA 0i5700 3. References /I/ Krauth, J.: A Locally r.csz powerful tied rank test in a Wilcoxon Situation; Ann, Math. Statist. 42, 1949 - 1956, 1971 /2/ Witting, H.: Mathematische Statistik; 2. Aufl. Teubner Stuttgart 1974; 173 - 177 /3/ R. Peto, M. C. Pike, ?. Amitage, N. E. Breslow, D. R. Cox, S. V. Howard, M. Mantel, X. McPherson, J. Peto and ?. G. Smith: Br. J. Cancer 35, l - 39, 1977 /4/ Finney, D.: Probit analysis, Cambridge University Press, 1971 n ,47 rt Louisiana Parish. Ca CMA 015701 ; t Bosa v . ; 1n u . Jn . , i.o. ifO-4837 14th Judif'i'i] ni.'itri -t Court Calcasir>u Parish, Louisiana Group Animals at start VII ! VI V IV bis IV III II I 90 100 30 120 30 30 30 60 male Alive animals after (week) 12 27 ' 36 55 Animals at start 87 C 80 C 73 C 99 C 92 C 80 C 30 30 28 120 117 113 30 29 28 29 27 26 23 x X 21 x x 18 x X 9 xX 6xx 5xX 70 C 90 56 C 10O 25 30 96 120 21 17 * 30 ' 30 12 X 30 1 XX 60 female Alive animals after (week) 12 27 36 55 89 C 88 C 86 C 85 C 98 C 97 C 92 C 73 c 30 30 30 26 119 118 118 112 30 30 29 26 29 28 27 ) 4 X X 30 28 26 1 3 X X 57 53 * A 46 x x 28 X X Table 24: Survival rates after 12, 27, 36 and 55 weeks after start of treatment (exact Fisher Test,one-sided) n > LI OSi w p a ris h , Louisian* r* opIDPW--* .1 CPD r* j male female Group VII Animals at start Lbfe time Mean Stand.Dev. Animals at start Life time Mean Stand.Dev. 90 70.7 C 22.58 90 81.0 C 18.51 VI IOO 75.4 c 22.08 100 83.1 C 20.32 V 30 83.7 x * 15.77 30 87.5 * 12.97 IV bis 120 75.0 15.85 120 84.3 17.41 IV 30 79.6 * 17.03 30 90.0 x * 18.40 Table 25:Mean and Standard Deviation (in weeks)of life time (Test of Krauth, one - sided) n 3>: 0H1 0X1l ow TMiI r,) o -i [. li 1 01 ^u<.0 ii S c+ n 33>; O LNOIl C O N F ID E N T IA L male female Group VI V IV VI V IV Animals at start IOO 30 30 IOO 30 30 interval N n.ma n.pa Mean ( 1, 89) 63 19 O0 .2 7 x 61.5 74.9x 20 0 3 71 .lx < 1, 89) 54 15 2 4* 43 69.0 77.5x 13 2 1 73.1 interval N n.ma n.pa Mean (90,150) 37 11 O0 1 4x 99.0 98.6 10 O 4x 96.7 (90,150) 46 15 17 O22 O O 6x 99.6 97.4 102.9 Table 26: Mean of life time and tumour incidence for MA and PA for special intervals; N ist the number of animals died in the interval; N.MA (respectively N.PA)is the number of animals with MA (respectively (Lxact Fisher test one-sided for tumour incidence, Test of Krauth one PA) s tiled for mean) male female i | Grouo VII IV bis VII IV bis Animals at start interval N ` ; N.MA 1 1 N.PA ! I Mean i1 I interval 1N N. MA 90 120 ( 1, 71) Hin O 0 1 SO. 8 3 60.0 x (72,150) SO 69 00 90 120 ( 1, 71) 25 31 ! ! | 03 0 60.4 2 63.1 ! : | (72, 92) 41 56 05 ! ; l 1 ! 1 ) j 111is la n a CMA 015705 N.PA Mean 2 86.7 13 * 1 86.1 80.4 3` l 84.5 x i interval N N.MA N.PA Mean : 1 11 1 i 1 i 1 ! (93,150) i ! 24 l * :i 1 1 i1 ! 1 103.7 33 4 6 103.8. ! ; ; 1 Table 27:Mean of life time and tumour incidence for MA and PA for special intervals; N is the number of animals died in the interval; S.MA (respectively N.PA) i3 the number of animals with MA (respectively PA) (Exact Fisher Test one-sided for tumour incidence, Test of Krauth one - 1 4 th Ju - ' C a lca sie u Paris'; Group Animals at start male 0 N.TU N.MA N.PA female Animals at start N.TU N.MA N-PA VI IOO 7 0 3 IOO 15 2 4 V 30 11 X 0 11 X 30 15 x 6 x 3 IV 30 11 X O 7x 30 17 x 4 * 7 x VII IV bis 90 120 70 43 x 1 3' 16 x 90 120 12 1 2 40 x 12 x 11 * Table 26: Tumour Incidence for HA and PA at the end of experiment; N.TU (respectively N.MA, N.PA) is the nunher of animals with tumours (respectively HA, PA) (Exact Fisher Test, one - sided). o3>: o tfl & 1 | Group i 71 i V :v mala ! female Animals at start N. V!A N. PA Animals at start n.ma n.pa 100 03 100 2 30 0 11 X 30 6x 3 30 0 7 x 30 4 7 * VII IV bis .-.90 120 03 90 1 16 * 120 12 12 * 11 Table 29: Tumour incidence for and PA at the end of experiment; N.MA (respectively N.PA) is the number of animals with MA (respectively PA) (Logrank test) Table 30 GflCLP : VI t SURVIVRL-flflTt tVCCl f*V i*;*3i ; s r <rs*r ,31 COrriD'iSNI 1AL Subject to Protect'." - c.^r Boss v Crt: 0 , !q. ^D-433Z 14th : ;h". h "^urt Calcasieu Parish, Lou^Lan* CMrt 015708 Table 31 CrOCP : VI SURVIVAL-RATE (VQD =N(1-^.5 -r 5T<SHT -11 ;C,s3 CONFIDENTIAL Subject tc ?:':" Ross v C:r,; - o 14th a Xalt . 1 r-.-i ' L -c . ;I y > in^ CMA 015709 Table 32 CflCL'P : 7 i SL'flVlVRL-RflTE CVQC1 Hfll_ Z*R:NZ<CK?^rtL'tJt-*M*ia?tT:?T3*7-eSs*rr ^exsj t,: .;33s3 CONFIDENTIAL Subject -Boss v i-iMa * Car"'"'. ' Pnt--' - - -- ' J'-r m !37 z ;.r,i CHA 015710 able 33 CHCUP : V SUHVIVfiL-RflTE (VCC) F"AL 'I'l: **n eni^.3 ar jrar i, -0i ,M CJ Su^c\ ' Boss v ' n. ,37 .na CMA 015711 Table 34 GRCwP : I / SUflVlVSL-r>fiTc (VGCJ :*1T -iHli snifsts -t s;s*r i s m .^1 ' 3Ur1 oasiana CMA 015712 Table 35 SURV[VPL-RfiTc CVCC1 GflCCP : [V FMfl_ >!*i; ;r i:ar -UAV , !;Js CO^IS'- Sub i- Bess 7 1` Cal': \o eu Parish, n ;a37 irt LouislahE CMA 015713 Table 36 GflCUP : V11 I SURViVflL-PRTE (VQCJ R_ ,:.w esr:*<if Kt*SJ ,> i j all^.S IT }T5*T , *J Eos_5_ V j}: " 44 - - I Cal isieu Parish. "i 1331 Hour6 Louial4flft CMA 015714 Table 37 CPCLF : VI; SURVIVai-RfiTE CV0C3 ' FSMfiU l :5C*Nrar*^ ppm ,3 *M**T'-5Nf *;* . % wtw^.3 if S7z*T f 93 ^ ^ - r -l "f m Y * Tj Suu - * . ' ' rc* in Boss 7 Conoco, I", ; . Co. CO* 433? 14th Judicial District Court Calcasieu Parish, Louisian* CMA Oi*"15 Table 38 CPCLP : [V 315 SUflVlVGL-flflTa CVCC) .'`Sit r f:s.'eiriv`*;tsj"`;Tjnv:tNr*f*i i. 15i :ri T VTj cosfB?- Sub3ect,, _ , .1 - in i. -i$34 l-itb p.-iju, '- Calc-VJ ? CMA 015716 Table 39 SURVlVfil-RATc tVDCJ caaf : IV 315 r*.VCLi * *'.MT 'eST;***Mt anc**.! at 3:r , ,3 , i2g 'T 1^- _ yi V * is ' co^ $-X-J Vo^3 '.<n 3 Im ^ar CMA 015717