Document ZBQxN0x3V7zrJj4g4nz4358G7

BOB WEST ASSOCIATES, INC. Uch'nfipc ^fivgu/afc nj n men 4//- %'a u> < ^'/wmicnh P.O.BOX 2001 STAMFORD, CONNECTICUT 06906 203-322-4466 CONFIDENTIAL PRIVILEGED Review and Audit of IBT No. 663-03222, Chronic Vapor Inhalation Toxicity Study with Vinyl Chloride (Ethylene Derived) in Albino Rats, Albino Mice and Golden Hamsters for the Manufacturing Chemists Association The review and audit of this study was conducted at IBT's offices in Northbrook, IL, April 14 to April 18, 1980. The review and audit on this study concentrated on the con duct and monitoring of the exposures. However, attention was also devoted to the source of the animals and body weight data as they affected fate and mortality. In addi tion, the review and audit included inspection of hand written observation books reflecting exposure concentrations, as well as gas chromatographic charts where they were avail able, hand written and computer body weight printouts, hand written mortality logs, hand written observations and cor respondence. CORRESPONDENCE, PROTOCOL AND TIMETABLE The timetable indicates the exposures were to be ini tiated on 9/10/73 for the T-I, T-II and T-III groups and on 10/1/73 for the T-IV groups. The termination of the exposures for the first three groups was to be at 9/8/74 and for the T-IV group at 9/27/74. The animals were then to be held for a year prior to terminal sacrifice. A letter dated June 15, Pth 0001297S CONFIDENTIAL IBT No. 663-03222 -2- PRIVILEGED 1973 from Dr. Torkelson of Dow reflects a site visit was made by the MCA Task Force, visiting the Decatur research labs of IBT where the study was conducted. Questions were raised about the adequacy of equipment, the exposures main tained, the exposure concentrations, the question of contam ination and record keeping. A major concern was raised about the adequacy of the analyses and record keeping. Hand written notes alongside the margin of this letter indicate that certain corrective actions were taken at IBT. I was unable to tell whether there were any other documented site visits. In addition, a progress report issued October 13, 1973 after about a month of exposure, reflects adjustments that were made to compensate for the issues that were raised during the MCA site visit. ANIMALS, HOUSING AND HUSBANDRY The animals for this study were supplied by the Charles River Breeding Laboratories. Unfortunately, we are not able to identify specific animal receipts with this project. However, we were able to identify from September invoices receipts of hamsters as replacements without charge for ani mals that were dead on arrival in the August shipment. We were able to identify an additional receipt of hamsters in early November, 1973, two months after the study started, which indicated 130 males and 130 females were received at that time. BOB WEST ASSOCIATES. INC . DTH 000012976 CONFIDENTIAL 1ST No. 663-03222 -3- PRIVILEGED Inspection of the body weights early in the study in dicates despite the report not reflecting the mortalities that there was a rather high mortality in the hamsters early in the study. At four and five months there were extensive replacements of the hamster groups. The animals were ear punched and the toes removed so the animals could be individually identified. This was to allow for identification for individual body weight determin ation and identification of when the animals died, because the animals were group housed in groups of 3-5 in stainless steel cages. Inspection of some of the hand written notes shows the layout of the rats in the rack and indicates that extra animals were to be spaced evenly throughout the rack. In addition, another hand written sheet shows the arrangement of the gang cages in the chambers. There were 18 gang cages, 8 rats each, to equal 144 rats. However, the protocol only calls for 100 rats at each level. This procedure allows for ample opportunities for replacement and substitution. Despite the fact that the animals were identified by ear punching and toe removal, most of the time the cages were not weighed individually but rather by the cage, e.g., 3-5 animals. Numerous hand written notes showing the arrangement of some of the animals in the racks reflect the animals excaped or were cannibalized, along with mortality dates for the remain ing individual animals. BOB WEST ASSOCIATES. INC. DTH 000012977 CONFIDENTIAL IBT No. 663-03222 -4- privilegEd VINYL CHLORIDE, SOURCE, IDENTITY Vinyl chloride (ethylene based) appears to have been supplied as, e.g., containers in excess of 500 pounds, by PPG Industries, Industrial Chemical Division, Lake Charles, LA. A letter from W. B. Braybill of PPG to Dr. John Goode of Bio-Test in Decatur indicates that one cylinder would be required per month. Heel sample analyses from the Bio-Test residues were also performed by PPG at the Lake Charles, LA operation and those that were available appeared to be in good agreement with the new sample analyses supplied to BioTest. Unfortunately, there was no inventory log at IBT for vinyl chloride. PPG personnel should be asked for account ability of the vinyl chloride sent to IBT. In addition, they would also have records of the returns for heel analyses. GENERATION OF VINYL CHLORIDE VAPORS The vinyl chloride supply tanks were kept in an explos ion room. The material was piped to the test chambers through thin-walled stainless steel tubing.. Before the thin-walled tubing delivered the vinyl chloride to the chambers, it was connected to a micrometering valve and a glass rotometer to control and monitor the supply of vapor to each chamber. Vapor-air mixture entered the top of the inhalation chamber, circulated through the chamber and was exhausted at the bot tom. Unfortunately, we were unable to find a daily log for BOB WEST ASSOCIATES, INC DTH 000012978 CONFIDENTIAL IBT No. 663-03222 -5- the weighing or measuring of the vinyl chloride that was dispensed during each daily run. CHAMBER MONITORING There are records of standardizations being conducted in September of 1973, covering the range from 20 to 100 ppm standardization and from 50 to 400 ppm for the 200 ppm standardization and from 400-4000 ppm for the 2500 ppm level. Standardizations were performed at approximately weekly in tervals prior to that point where the automatic sampling system was hooked up to the on-line gas chromatograph record er. The initial samples from the chambers for the first part of the study (six weeks) were obtained by use of grab samples which were then analyzed by the gas chromatograph. We were unable to find any record of daily sample analyses being taken in the research books for the month of October, 1973, the second month of the study, other than ranges and mean concentrations. There also was no evidence of chamber temp erature' or humidity being recorded. General Observations The following observations were obtained by inspection of research log books which present means and ranges of exposure concentrations, time interval sampling for the gas chromatographic concentrations and charts, as well as the hand written observations in the research log books. BOB WEST ASSOCIATES. INC'. DTH 000012979 CONFIDENTIAL IBT No. 663-03222 -6- PRIVILEGED 1. Most of the time the chambers were not on for six hours. 2. Most of the time the chambers were not even monitored for five hours, in some instances they were only moni tored for one to two hours. 3. In some instances there was selected data for the daily exposure concentration, e.g., values on the low side may have been eliminated or values on the high side may have been eliminated. 4. Periodically, there were mechanical problems associated with the rotometer, e.g., broken or clogged or unclean lines. 5. Some days there was no exposure because of equipment failure or TV filming or only a one-hour exposure was conducted. 6. There was evidence that the chambers did not have an adequate clearance time, since measurements made 45 minutes after termination of the exposure still showed rather high levels of vinyl chloride present. 7. There was evidence of contamination in the solvents room and the pilot plant which were in close proximity to the exposure room. 8. Some days there was no analytical data recorded for the exposure concentrations. 9. In some cases, the research log book showed means and ranges for exposure concentrations, with no raw supportive BOB WEST ASSOCIATES. INC'. DTH 000012980 CONFIDENTIAL IBT No. 663-03222 -7- PRIVILEGED data shown for periodic sampling with the gas chromato graph. 10. In some instances, the 50 ppm nominal concentration may have reached a concentration as high as 100 ppm and, in one instance, as high as 400 ppm which is not reflected in the means or ranges presented in the interim report. 11. In some instances, the control chamber indicated levels of vinyl chloride in a range of 5-11.5 ppm. 12. In September of 1974, near the end of the year of expos ure, there was some research book data reflecting exposure concentrations but no supportive gas chromato graphic data. 13. In some cases there is no data available for one or two levels out of the four exposure chambers being run. 14. There were periodic problemswith the gas chromatograph itself, either with the Ignitor, Detector or the Recorder. 15. In some instances there were means and ranges presented in the research book but no interval data were available. 16. In some instances, the chamber may have been on for four or five hours yet gas chromatographic readings were only taken for two hours. 17. There were a number of instances where the data presented in the research book do not agree with the data shown on the gas chromatographic chart. BOB WEST ASSOCIATES. INC. DTH 000012981 CONFIDENTIAL IBT No. 663-03222 -8- PRIVILEGED Specific Exposure Observations The following notations were associated with the spe cific exposures and may also be used together with the inhalation chamber concentrations presented in Table I. The exposure concentrations presented in Table I are those presented by IBT and one can see with the use of these spe cific notations that the values presented in Table I are not an accurate reflection, in many instances, of the exposure concentrations or are based on minimal supportive analytical data. Exposure Number 5 20 & 35 53 56 80 83 Comment The values for the T-I mean and the range for the raw data in the report do not agree. In the daily intervals, there were problems with sampling and the curve does not appear to be of value. 200 ppm equaled 400 ppm. The gas chromatographic charts are no good and the research book concentration is not shown or recorded. The gas chromatographic chart agrees with the data in the research book. There are no gas chromatographic charts but there is data in the research book. BOB WEST ASSOCIATES. INC. DTH 000012982 CONFIDENTIAL I3T No. 663-03222 -9- PRIVILEGED Exposure Number 85 86 108-110 134&135 141 144 145 Comment There are no T-I or T-II data on the chart or in the book. T-III data is present in the research book but not on the gas chromatographic chart. The gas chromatographic chart and the data in the research book agree but the exposure concentra tions are on the low side. The gas chromatographic charts do not match the data in the log book. Observations were noted that some of the data was off the GC chart and there were some operator errors and missing data. There were no GC charts to go with the data pre sented in the research books. No charts available for the exposures 110-136. A line was drawn through the first hour values and apparently discarded. There are gas chromatographic charts and research book data available but another notebook says that exposures were not run. In addition, the control line shows contamination. A notation was made that some of the readings were off the chart -- wide excursions on both the low and high sides. In addition, the controls were contaminated. BOB WEST ASSOCIATES. INC. DTH 000012983 CONFIDENTIAL IBT No. 663-03222 -10- PRIVILEGED Exposure Number 146 150 152 158 159 165 Comments Some of the data from the charts has been dis carded. The T-II value at the first sampling interval does not agree with the data presented in the research book. The negative control line also shows con tamination. The T-III value is on the high side and reaches 3420. A broken rotometer on line T-I. The values for the T-III line indicate they are not working and they have been deleted. The control background is high. Selected data has been used for the tabulated data presented. Analytical samples are present for four hours. There are missing data when one compares the GC charts versus that present in the research book. The chambers were only on for two hours. There are no exposure concentrations for T-II, T-III and T-IV at the first sampling. There are incomplete values for the second sample at two hours. The chamber was operated for 4 1/2 hours. Zero time was taken ten minutes after the start of the exposure, at which time the gas chromatographic readings were taken. Environmental samples on the BOB WEST ASSOC IATES. INC. DTH 000012984 CONFIDENTIAL I3T No. 663-03222 -11- PRIVILEGED Exposure Number 165 (cont'd) 166 169 180 196 197 Comments roof appeared high. Hood efficiency, where the syringes were cleaned was poor. Vinyl chloride was still detectable in the T-IV chamber 45 min utes after the exposure was terminated. The solvent room and the pilot plant both showed vinyl chloride contamination. The first reading on the gas chromatographic chart is 3:55 pm. The readings continue until 4:40 pm at sharply lower concentration. The chamber was turned on at 10 o'clock in the morn ing. Sampling was only conducted for 4 hours. The T-III value was 3710 ppm. For the T-I and T-II lines, there was no sample taken for the second sampling interval or the fourth interval. Basically no analytical data on exposure concen tration. Exposures were sampled for five hours. Selected data in the daily range and no noon samples. The control line was contaminated. The chambers ran for five hours and GC data are available for three. BOB WEST ASSOCIATES. INC. DTH 000012985 CONFIDENTIAL IBT No. 663-03222 -12- PRIVILEGED Exposure Number 201 206 210 218 224 225 Comment A seven-hour exposure is indicated as having been run. The sampling for GC analysis was run for somewhat less than five hours. Selected data has been taken at the initial reading and then spor adic data is missing throughout the run. The chambers were not monitored and no data are available. No sampling. Exposures were only on for one hour with a notation of TV filming. There was one reading for each line for the first two hours, obtained manually. Half of the samp lings were done on automatic, but two, rather than three, samples were obtained. Notation is made of CBS filming tomorrow; expos ures were run for 4 1/2 hours. The line control had a reading of 6. The GC chart shows no agree ment with the research book, which is sparse. However, it does show T-III range from 3-320 ppm. The T-I 50 ppm range equals 445 ppm. No gas chromatographic charts to support the mini mal data in the research book. The chambers were turned on at 9:00 am and were off at 11:40 am. GC readings were taken only at 10:00 and 11:00 am. BOB WEST ASSOCIATES. INC. DTH 000012986 CONFIDENTIAL IBT No. 633-03222 -13- PRIVILEGED Exposure Number 228 231 263S237 238&239 243 244 251 255 Comment For T-I and T-II, there are missing data. For T-III, there are no values for the first and second samplings. One-third of the values which were on the low side were deleted from the calculation of the averages, one of which was off the graph. Se lected data is shown. The control shows a peak height of 7.5, indicating some contamination. The gas chromatograph was not working; no data but means have been reported for one value at one interval. The research book shows a mean and ranges but no supportive data were available. The line control shows a peak height of 8. Onethird of the data from the GC charts were not used in the calculations or averages in the research book. Samples are only shown for three hours. The GC chart data is sparse. The line control is shown as 6.5. The 50 ppm equaled 400 ppm and a notation of major problems. Air recirculated was causing high concentrations in the control chamber. The chambers were turned BOB WEST ASSOCIATES, INC'. DTH 000012987 CONFIDENTIAL IBT No. 663-03222 -14- PRIVILEGfeD Exposure Number 255 (cont'd) 257 258 Comment off. The control chamber had a level equal to 3.4 ppm of vinyl chloride. A notation of definite leakage into the room was recorded. Selected data only for two hours of the exposure was used'. The T-IV chambers were turned on two hours after the rest of the chambers. The line control shows vinyl chloride contamination. One of the 50 ppm values is equal to at least 100 ppm. In addition, one-third of the values were unavailable and se lected data were used. The first exposure concentration readings were taken at 1:15 pm and also at 2:15, despite the fact the chambers were turned on at 11:30 in the morning. Most of the data were incomplete. This appeared to be the last exposure where gas chromatographic chart data were available.. The research book has data for exposures 259, 260, 261, 262, but there are no sup portive gas chromatographic data. BOB WEST ASSOCIATES, INC. DTH 000012988 CONFIDENTIAL IBT No. 633-03222 -15- PRIVILEGED BODY WEIGHTS Body weights were available, either as hand written calculations or as computer printouts. Unfortunately, many of the computer printout body weight tabulations reflected the animals being weighed on a gang-cage basis. Therefore, it becomes more difficult to determine substitutions. A tabulation beginning with month 5 and proceeding through month 13 is shown below for the body weight data. This com pares the data presented in the interim IBT reports versus that obtained when one inspects the computer printout itself. These data have only been examined for the T-III group. MEAN BODY WEIGHTS FROM PRINTOUTS VINYL CHLORIDE RATS, T-III - 2,000 PPM Month Male Female Month Male Female 5 Report 549 321 Printout 550.5 312 9 Report Printout 681 359 627 359 6 Report 567 329 Printout 567.3 339 10 Report Printout 663 356 629 356 7 Report 565 Printout* 714 691 702 606 346 436 11 Report Printout 12 Report Printout . 696 680 736 718 374 374 378 378 8 Report 585 Printout** 581 359 359 13 Report Printout 740 420 729 420 *A11 values shown on printout **Subtracted 329 grams from printout mean Inspection of these data indicates that during the seventh month there is a major difference between the body weights in the printouts and those presented in the final report, suggesting replacements. BOB WEST ASSOCIATES. INC. DTH 000012989 CONFIDENTIAL IBT No. 633-03222 -16- PRIVILEGED MORTALITY AND FATE Hand written sheets reflecting tabulations of mortali ties are available. These sheets bear various dates at periodic intervals. Additional tabulation reflects animals that are missing or cannibalized, based upon the gang hous ing. However, there are numerous errors present and inconsis tencies in these tabulations. A chart of missing rats, pre pared near the end of the third month of the study, shows numerous animals not accounted for in Dr. Shah's tabulation. These differences are as follows: Untreated control male rats - 2 animals missing; none shown on Dr. Shah's tabulation. Untreated control females - 3 missing; none shown on Dr. Shah's tabulation. T-I male rats - 7 missing; 1 shown on Dr. Shah's tabulation. T-II male rats - 6 missing; none shown on Dr. Shah's tab ulation . T-III male rats - 4 missing; none shown on Dr. Shah's tab ulation. T-l female rats - 6 missing; none shown on Dr. Shah's tab ulation. T-II female rats - 2 missing; none shown on Dr. Shah's tab ulation. T-III female rats - 4-5 missing; none shown on Dr. Shah's tabulation. BOB WEST ASSOCIATES, INC'. DTH 000012990 CONFIDENTIAL IBT No. 633-03222 -17- PRIVILECED There are numerous evidences of errors in the animal numbers or the animals being missexed,e.g., animal 751 female shouldbe 638 male in the T-III group; animal 787 female shouldbe male 691 in the T-III group; animal 788 female shouldbe 692 male in the T-III group; animal 689, T-III male rat should be 686; 605, T-III female rat, is a male. Autopsy sheets that reflect extra animals were avail able . A mortality tabulation prepared from the animals avail able in each group, as determined by the body weight print outs, beginning with month five and continuing through month 13 is shown in the accompanying Table. You will note a sub stantial difference in the numbers available as tabulated between the printout and that presented in the report for the male T-III group, month 7, 12 and 13. Similarly substantial differences in the females in months 12 and 13, as well, also suggests substitution of the animals was occurring. Close attention was paid to determining the specific fate of the five animals examined in detail by Dr. Bussey in the T-III male group, as reported to have brain tumors. Inspection of the hand written post mortem tabulations by IBT staff, as opposed to the data presented on the autopsy sheets and the mortality data presented in the final report indicates substantial discrepancies. BOB WEST ASSOCIATES, INC. DTH 000012991 CONFIDENTIAL IBT No. 633-03222 -18- PRIVILEGED Animal #641, T-III male, was found dead at 369 days, according to the report. Hand written P.M. tabulation sheet shows the animal to be dead by 8/6/74, which is about 330 days into the study. The necropsy sheet shows the animal's date of death as 9/3/74 (359 days). These differences have not been explained. In the case of animal #649, the necropsy sheet and the report shows that the animal died at 402 days. The sheet presenting post mortem tabulations, dated 9/11/74, however, shows the animal as being dead. This date would be approxi mately one year into the study. Animal #664 was reported as having died at 373 days. This animal is shown as being dead 7/17/74 on the necropsy sheet and as being dead on a hand written tabulation or. 8/9/74. Therefore, the 373 day mortality presented in the report is incorrect and was probably 311 days. Animal #672, shown in the report as having died at 310 days, is also shown on a post mortem mortality tabulation dated 7/26/74 as being dead by that time. A necropsy date of 7/16/74 for this animal is noted. Therefore, the fate of this animal appears to be correct. In the case of animal #667, the report indicates days on test as 383 but the necropsy sheet is dated 6/27/74, re flecting less than a year on test. A post mortem log dated 6/26/74 also shows the animal as being dead. Therefore, the date presented as days on test in the report is in error. BOB WEST ASSOCI ATES, INC. DTH 000012992 CONFIDENTIAL IBT No. 633-03222 -19- PRIVILEGED CONCLUSIONS An intensive audit of certain aspects of IBT No. 633-03222, chronic vapor inhalation toxicity study with vinyl chloride, devoted to inspection of exposure concentrations and pertin ent body weight and mortality logs indicates the following: (a) There are numerous evidences of replacement of animals in this study. There appears to be an exceptionally high replacement of the hamsters. Also, because of the dif ferences in body weights and mortalities noted in the T-III rats, it is possible that there was major substi tution of the animals between the sixth and seventh months of the study and perhaps at 12 and 13 months. There are evidences of extra animals being available for the various species in the study. The accountability for the animals and the mortalities show substantial discrepancies. (b) The monitoring of the exposure concentrations and the conduct of the exposures shows poor technique and selec tion of data in many instances but does seem to indicate that a three-level study was conducted, even though the swings in exposure concentrations periodically may have been 50-100%. The instances of actual overlap between exposure concentrations at the T-I level and the T-II level were relatively infrequent and there do not appear BOB WEST ASSOCIATES. INC'. DTH 000012993 CONFIDENTIAL IBT No. 633-03222 -20- PRIVILEGED to be any real instances of overlap between the T-II and T-III levels. (c) The animals reported to have brain tumors indicate discrepancies as to their actual dates of death versus those reported in interim reports and in some cases on the necropsy sheets. In summary, the study was conducted in an extremely sloppy fashion and one would have to look at any of the re sults more from a qualitative standpoint than a quantitative standpoint. Bob West, Ph.D. BW/am Date: May 10, 1980 BOB WEST ASSOCIATES. INC'. DTH 000012994 DTH 0 0 0 0 1 2 9 9 5 c to s03 0) S-l * 0) a > 03 H e1 to a X Eh 0) 3> cc 0 (0 ViH X 0 o T3 C to *-N 0 03 U 0> H s *-l c to s0) M o O CM co iH in o a in co ID c- CO O in r-H o CO o o CO o CM pH O in in h VO o CO in CO in nr ON O iH in CO ID o CM CO nr tN CO m in CM in nr o m nr H in 00 in m c* in in in ID in CM in in in CM CM CM CM iH CM (N fN (N n CM CM CM CM CM CM CM CM CM CM fM CM CM fM tN CM <N CM u0 CD c 1 Me a ooOo O O o Q H Eh rH iH nr o CM CM CM CM CM CM pH X3 0 rH to 0 ID ID iH pH nr r- c- rH nr c* rH r- nr CM r* 3 CM CO CM m ny CM CM CO CM CO CM CM CM CO CM co CM CO CM CO CO CM CO CM CM co C 0 pH < te. 4J to D y-v c 10 a O Ooo o >i 03 X 4J c E a. 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HJ HJ 3 XI 0 r- G 0 CN rH 4J 4H G 0 Vh 4J cC G0 00 Ec 0 G0 r> 4J H H 1 a0 E-t U * 0 0X r- X 4J r- -P o CN i HH 0 o 0X r- 4-> 0 rH G3 X G -P w *H G 0 EH iH 0 m 0G Hs c 00 J3 C -H 0 -u V H G 0 P >H 4J u x> U 0c 0 a0 cu 0 U 0 CG 1h rH O U >1 0 iH 10 to 0 0 -H 3 iH X 0 Cm +J H > 0 rH 4J u C H CH O 2 He * O CP 33 rH O o o o % TABLE II IBT No. 663-03222 VINYL CHLORIDE RATS - SURVIVAL T-III - 2,000 ppm Month 5 Report Printout 6 Report Printout 7 Report Printout* 8 Report Printout 9 Report Printout 10 Report Printout 11 Report Printout 12 Report Printout 13 Report Printout** Male 97 96 95 96 94 89 or 92 90 89 84 82 76 74 66 62 42 37 27 23 Female 96 95 96 94 95 92 92 90 85 81 72 68 ' 48 46 34 15 (?) 17 5 (?) * None of #'s or weights agree with report ** All over the lot, 5 body weights shown DTH 0000129S7