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CALIFORNIA DEPARTMENT OF PESTICIDE REGULATION
MEDICAL TOXICOLOGY BRANCH
SUMMARY OF TOXICOLOGY DATA
DAMINOZIDE
(Studies with UDMH included)
Chemical Code: 000007, Tolerance # 246
SB #: 136
July 30, 1986
Revisions 7/16/87, 12/23/87, 10/27/88, 4/20/89, 11/29/89, 3/16/90, 11/16/90, 8/1/91
I. DATA GAP STATUS
Combined, rat: No data gap, possible adverse effect (oncogenicity)
Chronic toxicity, dog: No data gap, no adverse effect
Oncogenicity, mouse: No data gap, possible adverse effect
Reproduction, rat: No data gap, no adverse effect
Teratology, rat: No data gap, no adverse effect
Teratology, rabbit: No data gap, no adverse effect
Gene mutation: No data gap, no adverse effect
Chromosome effects: No data gap, no adverse effect
CDPR MEDICAL TOXICOLOGY DAMINOZIDE, SUMMARY OF TOXICOLOGY DATA T910801
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DNA damage: No data gap, no adverse effect
Neurotoxicity: Not required at this time
______________________________________________________________________________
Toxicology one-liners are attached.
In record/document number identifiers for one-liners below:
** indicates an acceptable study.
Bold face indicates a possible adverse effect.
File name: T910801
Revisions by Martz, 7/16/87; Gee, 12/23/87; Kishiyama & Davis, 10/27/88; Gee, 4/20/89; Silva,
11/29/89; Gee, 3/16/90; Aldous, 11/16/90; Kellner, 8/1/91.
Rectified with Library printout through volume 051, Record No. 097254. All relevant records
available as of 7/25/91 have been examined.
CDPR MEDICAL TOXICOLOGY DAMINOZIDE, SUMMARY OF TOXICOLOGY DATA T910801
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II. TOXICOLOGY ONE-LINERS AND CONCLUSIONS
COMBINED, RAT
NOTE: CDFA has considered the rat chronic and oncogenicity study requirements filled as of
October 1988 reviews by B. Davis. No single study was considered acceptable at that time,
however Dr. Davis considered the collective data to be adequate for oncogenicity and chronic
study data requirements, particularly on the strength of daminozide study 032:070327, which
was faulted only on the basis of dose level justification. An earlier NCI daminozide study
(005:026447) differed from current guidelines in several respects, but also provided useful
information. Since the 1988 CDFA reviews, the UDMH study was completed at IRDC (043:085116).
None of these studies provided definitive evidence of dose-related tumor effects. Each of the
three studies used Fischer 344 rats. The NCI study (026447) provided the strongest evidence
for a treatment effect on uterine adenocarcinomas. Both of the other studies provided weak
additional support for a treatment effect for such tumors. In addition, both daminozide
studies indicated low levels of uterine leiomyosarcomas as a possible treatment response: this
was not indicated in the UDMH study. A possible increase in hepatocellular tumors was
observed in females in the UDMH study only. These observations suggest marginal evidence of
oncogenicity in the rat. Aldous, 11/7/90.
246-032, Parts 1-6 070327 Johnson, D.E., "Two Year Dietary Toxicity and Oncogenicity Study in
Rats" (IRDC, Study No. 399-055, 8/25/88). Daminozide (Alar Technical, Lot No. M074023NB,
99.8%; UDMH content = 29 ppm) fed to Fischer 344 rats (60/sex/group) at 0, 100, 500, 5,000,
and 10,000 ppm for 2 years; interim sacrifice of 10/sex/group at 1 year. POSSIBLE ADVERSE
EFFECT (presence of comparatively uncommon uterine tumors: incidences in controls through
increasing dosage groups were 0, 1, 2, 1, and 1 for uterine adenocarcinomas; and 0, 0, 0, 2,
and 1 for leiomyosarcomas). A previous study (1978 NCI study at Litton Bionetics; Record
026447) also noted increases in these cell types. Chronic toxicity NOEL > 10,000 ppm (HDT);
UNACCEPTABLE-an MTD was not achieved. Davis, 10/20/88; 1-liner incidence details added
11/7/90 by Aldous.
CDPR MEDICAL TOXICOLOGY DAMINOZIDE, SUMMARY OF TOXICOLOGY DATA T910801
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246-035 067727 Supplement to 070327. Contains pages missing from the initial submission -
265, 273, 481-490, 492-498, 514, 515, 645 and 653. No worksheet. Gee, 3/13/89.
246-039 072778 Supplement to 070327. Contains replacement pages for tables containing
data in which one 0 ppm rat was incorrectly listed as having "malignant lymphoma, lymphocytic"
but should read "mononuclear cell leukemia." No worksheet. Gee, 3/13/89.
246-017, 050911: "Two Year Dietary Toxicity and Oncogenicity Study in Rats" (12 Month
Interim Report, Study #399-055) - see # 070327.
246-018, No record number; 3/11/87 letter from registrant with brief summary of new combined
rat study; Contains summary of #050911 and the EPA submission date (listed as 8/15/88). Martz
6/22/87.
246-037 072994 Partial duplicate of 032:070327, above.
246-013 036932: Oser, B.L. "Chronic (2-Year) Feeding Studies With B-995 in Rats and Dogs"
(FDRL, 11/15/66) Daminozide (no purity information) at 0, 300, 1000 or 3000 ppm for 2 years
to 25 rats/sex/group; 37 rats/sex/control group; UNACCEPTABLE, NOT UPGRADEABLE: dose levels
not justified (3000 ppm max); too few animals per group (25); no dose analysis; inadequate
histopathology. NO ADVERSE EFFECT reported. Gee 11/26/85.
246-037 Documents submitted in support of the position that daminozide should not be
considered a carcinogen until the UDMH studies are completed and reported. Comments were
submitted by several consultants to the registrant and were those presented at a meeting on
January 26, 1989, with CDFA. Gee, 3/10/89.
246-038 A compilation of references supporting the comments of Dr. Edward Ilgren as
discussed at the January, 1989, meeting. These have not been individually reviewed for SB950
but are considered in the risk assessment process. Some reports contain record numbers while
others do not. Gee, 3/10/89.
CDPR MEDICAL TOXICOLOGY DAMINOZIDE, SUMMARY OF TOXICOLOGY DATA T910801
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CHRONIC TOXICITY, DOG
** 036, 042 071340, 075754 "One year dietary toxicity study in dogs," (IRDC, 399-066,
11/14/88). Daminozide, technical (purity = 99%; 29 ppm UDMH and 0.06 ppm NDMA) was fed in the
diet for one year at 0 (diet), 300, 3000 or 7500 ppm; 6/sex/group. No effect on any parameter
measures; which included body weight, clinical observations, ophthalmology, microscopic
examination. NOEL > 7500 ppm. Previously reviewed as unacceptable (Gee, 3/10/89). Upon
receipt and evaluation of information regarding dose justification, the study has been
upgraded to acceptable. Silva, 11/15/89.
246-013 036934 Oser, B.L., "Chronic (2-Year) Feeding Studies With B-995 in Rats and Dogs"
(FDRL, 11/15/66). Daminozide (no purity information) fed to 4 dogs/sex/group at levels of 0,
300, 1000 or 3000 ppm for 2 years; UNACCEPTABLE-needs justification of dose selection,
description of a.i. purity, more details of protocol, histopathology. NO ADVERSE EFFECT
reported. Gee 11/26/85.
ONCOGENICITY, RAT (DAMINOZIDE STUDIES)
246-005, 026447: "Bioassay of Daminozide for Possible Carcinogenicity" (NCI/Litton Bionetics,
1978) Daminozide (greater than 99%) fed in diet to Fischer 344 rats for 2 years at 5000 and
10,000 ppm; 50/sex/group in treated rats, or 20/sex/group in concurrent controls. Possible
adverse effect (presence of the following comparatively uncommon uterine tumors): incidences
in controls through increasing dosage groups were 0/19, 5/50, and 3/50 for endometrial
adenocarcinomas and 0/19, 1/50, and 3/50 for leiomyosarcomas. Identification of 2 of the 4
leiomyosarcomas was later challenged on re-evaluation of the slides [see record 940, below].
Historical control incidences were 2/220 for adenocarcinoma and 0/220 for leiomyosarcoma.
UNACCEPTABLE; CANNOT BE UPGRADED-only 20/sex in control groups; two doses (based on a
subchronic study) with no evidence of MTD; higher mortality in control males may have
confounded statistics. Second opinion by Martz: agree. The occurrence of rare tumor types
only in treated animals indicates toxicologic significance in spite of other deficiencies.
CDPR MEDICAL TOXICOLOGY DAMINOZIDE, SUMMARY OF TOXICOLOGY DATA T910801
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Gee 5/6/85, with second opinion review by Martz 4/14/86, revised 5/14/86, and 11/5/90 (Aldous,
no new worksheet).
CDPR MEDICAL TOXICOLOGY DAMINOZIDE, SUMMARY OF TOXICOLOGY DATA T910801
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246-006, 940 [in Appendix II] Supplemental to 005, 026447. "Daminozide Study Carried Out
At Litton Bionetics, Inc., Rat Study," no date. Second opinion diagnoses by Dr. Stan D.
Vesselinovitch, pathologist, of uterine tumors in Litton/NCI study discussed above. Dr.
Vesselinovitch and the NCI pathologist differed on the diagnosis of two (of 4) leiomyosarcomas
[Litton pathologist’s diagnoses of "leiomyosarcoma" were changed to "sarcoma" for one low dose
female, and to "neurilemoma" in one of three affected high dose females]. Diagnosis of the
remaining 2, both in the high dose group, remained unchanged. Although the differences from
control were not significant, a treatment-related effect still cannot be discounted, because
this tumor type is rare and was found exclusively in the high dose group. Endometrial
adenocarcinoma diagnosis remained unchanged, and the incidence was statistically significant.
CDFA review by F. Martz, 4/14/86 (no supplemental information review worksheet prepared).
246-006, 038449: Supplemental to 005 026447. "Uniroyal Summary and Review of Daminozide
and UDMH Oncogenicity Studies,"--Rat; no date; Uniroyal’s summary /critique of daminozide
rodent studies. CDFA review by F. Martz 5/14/86 (no supplemental information review worksheet
prepared).
246-006, 000937: Supplemental to 005, 026447. "Overall Incidence of Relevant Neoplastic
Lesions Identified in the Daminozide Studies," no date. Summary table. CDFA review by F.
Martz, 5/14/86 (no supplemental information review worksheet prepared).
ONCOGENICITY, RAT (UDMH AND OTHER SUBSTITUTED HYDRAZINES)
**246-043 085116 Goldenthal, E. I., "Two year oncogenicity study in rats", IRDC Study 399-
062, 9/28/89. Charles River Fischer-344 rats were supplied UDMH (major metabolite of
Daminozide) in pH-buffered drinking water at 0, 1, 50 or 100 ppm. Seventy/sex/group were
initiated on study. Of these, 20/sex/group were sacrificed at 12 months: the remainder were
maintained for 2 years. Purity of UDMH was not specified, however solutions were
satisfactorily assayed against a commercial standard. NOEL = 1 ppm (decreased water
consumption in females, possibly due to smell or taste aversion to test article). A practical
CDPR MEDICAL TOXICOLOGY DAMINOZIDE, SUMMARY OF TOXICOLOGY DATA T910801
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level for which no apparently meaningful toxicity is observed would be 50 ppm in males and
females. At 100 ppm there were modest, but consistent body weight decrements in both sexes,
in conjunction with reduced water consumption (especially in females). Possible adverse
effects were noted: a modest increase in incidence of hepatocellular tumors in 50 and 100 ppm
females, also the presence of two high dose females with comparatively rare uterine
adenocarcinomas, provide marginal evidence of treatment effects. Aldous, 11/16/90.
246-049 073813 A copy of R. A. Squire’s analysis of hepatocellular tumors and related
lesions in rat study 043:085116, above. The same analysis is found in part 6 of the final
report in Vol. 043.
ONCOGENICITY, MOUSE
**246-031, Parts 1-5 070326 Johnson, D.E., "Two Year Dietary Oncogenicity Study in Mice"
(IRDC, Study No. 399-054, 8/29/88). Daminozide (Alar Technical, Lot No. M074023NB, 99.8%:
UDMH content = 29 ppm) fed to CD-1 mice (50 mice/sex/group) at 0, 300, 3,000, 6,000, and
10,000 ppm for 2 years. This study did not demonstrate an MTD, however dose levels are
justifiable on the basis of earlier studies (see 005:026448). Brown pigmentation in livers of
6000 and 10000 ppm males suggested a modest treatment effect, with a NOEL of 3000 ppm for
males. Apparent NOEL for non-neoplasia effects in females was 10000 ppm. A "possible adverse
effect" was equivocal evidence of a high dose effect on vascular tumor (hemangioma plus
hemangiosarcoma) incidences in livers of 10000 ppm males and in uteri of 10000 ppm females.
Evidence of a pulmonary tumor effect was insufficient to warrant a "possible adverse effects"
call. (The 1988 CDFA review had considered the pulmonary tumors to indicate a "possible
adverse effect"). Acceptable. Davis, 10/20/88; Aldous, 11/7/90. [Note: high doses of UDMH
predictably elicit vascular tumors and pulmonary tumors. See individual 1-liners in the
"substituted hydrazines" section, below].
246-037 072993 Partial duplicate of 031:070326, above.
CDPR MEDICAL TOXICOLOGY DAMINOZIDE, SUMMARY OF TOXICOLOGY DATA T910801
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019, 057774: "Two Year Dietary Oncogenicity Study in Mice," IRDC, 10/30/86; one year
interim report cited in the letter of 11/24/86.
029, 065260: Interim pathology report for Record 070326. Davis 10/7/88.
018, No record number: Letter from registrant dated 3/11/87 with brief summary of new
studies in progress including the repeat mouse study [see 070326] Martz 6/22/87.
017, No record number; Cover letter from registrant dated 11/24/86: similar information to
that given above in 018. Martz 7/16/87.
246-005, 026448: "Bioassay of Daminozide for Possible Carcinogenicity"--Mouse, NCI/Litton
Bionetics, 1978; daminozide (greater than 99% purity) to B6C3F1 mice at 5000 and 10,000 ppm in
feed, 2 years. 50/sex/group, 20/sex/group for controls; ONCOGENIC EFFECT - hepatocellular
carcinomas in males, dose-related trend with significance at high dose level. Lung tumors
(alveolar/bronchiolar adenomas and carcinomas) in females, not significant by Fisher’s. Note
that lung tumors were also reported in Swiss mice by Eppley (see below, # 023502) and in CD-1
mice by IRDC (see above, # 070326). UNACCEPTABLE & CANNOT BE UPGRADED: dose selection (no
evidence of toxicity), too few controls, two doses only. Gee 5/6/85.
246-038 072541 Duplicate of 026447 and 026448 (above).
246-006, 038450: Supplemental to 005, 026448. "Daminozide Study Carried Out At Litton
Bionetics, Inc., Mouse Study," no date, Second opinion review by Dr. Stan D. Vesselinovitch,
pathologist, of selected liver tumors in male and female mice of the Litton/NCI study
discussed above. There were no substantial differences in tumor diagnoses between Dr.
Vesselinovitch and the NCI pathologist. Martz 5/14/86 (no supplemental information review
worksheet prepared).
CDPR MEDICAL TOXICOLOGY DAMINOZIDE, SUMMARY OF TOXICOLOGY DATA T910801
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246-006, 031139 & 038451: Supplemental to 005, 023502 & 026448. "Overall Incidence of
Relevant Neoplastic Lesions Identified in the Daminozide Studies" in mice, no date. Summary
comparison table. Martz, 5/14/86 (no supplemental information review worksheet prepared).
246-006, 945 & 946: Supplemental to 005, 023502 & 026448. "Uniroyal Summary and Review of
Daminozide and UDMH Oncogenicity Studies,"--Mice; no date; Uniroyal’s summary/critique of
daminozide rodent studies. Martz, 5/14/86 (no supplemental information review worksheet
prepared).
246-005, 023502: Toth, B., Wallcave, L., Patil, K., Schmeltz, I., and Hoffmann, D., "Induction
of Tumors in Mice With the Herbicide Succinic Acid 2,2-Dimethylhydrazide," Cancer Research 37,
3497-3500, from Eppley Inst., 10/77; Oral dosing in drinking water at 2% concentration,
equivalent to 3000 mg/kg/day, in Swiss mice, 100/sex in controls and 50/sex in treatment
group. Contained 0.002% UDMH (40 ppm), known to be tumorigenic in mouse lung (see #23503
below), equivalent to 6 mg/kg/day; ONCOGENICITY in lungs (primarily adenomas, with some
adenocarcinomas), blood vessels (particularly angiosarcomas, with some angiomas: these were
most abundant in the liver), and kidney (adenomas). Lung tumors probably due to UDMH
contaminant rather than test article itself. Treatment relatedness of kidney tumors is
subject to question (see below). Nevertheless, incidences of vascular tumors merit attention
in spite of scientific shortcomings. Study is UNACCEPTABLE for regulatory purposes due to
multiple deficiencies, such as grossly exceeding the MTD so that survival was severely
reduced: study CANNOT BE UPGRADED. Gee 5/6/85, with second opinion Martz 5/14/86.
246-038 072765 Duplicate of 005:023502, above.
246-006, 000941 & 000942: Supplemental to 005, 023502 above. "Daminozide Study Carried Out
at Litton Bionetics, Inc.," mice, no date. Second opinion diagnoses by Dr. Stan D.
Vesselinovitch, pathologist, of tumors in Eppley study discussed above. Dr. Vesselinovitch
and the Eppley pathologist differed on numerous diagnoses such that the incidence of vascular
tumors in liver was reduced (but still significant), and kidney tumor incidence was reduced
and no longer significant. Lung tumor incidence remained unchanged from original diagnoses.
CDPR MEDICAL TOXICOLOGY DAMINOZIDE, SUMMARY OF TOXICOLOGY DATA T910801
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The consultant audited the data files and found substantial evidence of poor study conduct.
Consequently, the results are subject to question. Nevertheless, the significantly elevated
incidences of lung and vascular tumors can not be discounted in spite of this study’s
deficiencies. Martz 5/14/86 (no supplemental information review worksheet prepared).
246-006, 000938: Supplemental to 005, 023502; "Development of Blood Vessel Tumors in Swiss
Mice (Eppley Subline) as Reported by Bela Toth," no date. Incidence of vascular tumors in
male and female controls from 18 references. Martz 5/14/86 (no supplemental information
review worksheet prepared).
ONCOGENICITY, MOUSE (UDMH AND OTHER SUBSTITUTED HYDRAZINES)
NOTE: Taken together, the two IRDC studies below (Volumes 44 and 48) show clear potential for
UDMH to elicit bronchiolar/alveolar tumors as well as vascular tumors (particularly
hemangiosarcomas, commonly observed in liver) in both sexes at or above 40 ppm. In addition,
the lung tumors were observed in 20 ppm females. Minimum doses associated with tumor
development approximated the MTDs in respective sexes. Although the "possible adverse effect"
of UDMH cannot be ignored, the findings below should be considered in the context of possible
excessive toxicity. Aldous, 10/22/90.
**246-048 090438 Goldenthal, Edwin I., "Two year oncogenicity study in mice", IRDC, 1/31/90.
CD-1 mice were supplied UDMH (major metabolite of Daminozide) in pH-buffered drinking water at
0, 40, or 80 ppm. Out of 90/sex/group originally assigned, 20/sex/group were sacrificed at 8
months and an additional 20/sex/group at 12 months: the remainder were maintained for 2 years.
Purity of UDMH was not specified. Possible adverse effects were noted: vascular tumors
(particularly hemangiosarcomas in liver) and alveolar/bronchiolar tumors (primarily adenomas)
were each elevated in both sexes at both dosages. The high doses (80 ppm) in both sexes
clearly exceeded the MTD, based on survival criteria. Reduced survival at the 40 ppm dose in
females may also have represented a treatment effect, hence tumor effects must be interpreted
in the context of high toxicity. Liver toxicity (and/or adaptive responses) was demonstrated
CDPR MEDICAL TOXICOLOGY DAMINOZIDE, SUMMARY OF TOXICOLOGY DATA T910801
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by hypertrophy, necrosis, and brown pigmentation in both sexes at both dosages. In addition,
an inflammatory response was evident in males at both doses. Serum chemistry findings of
elevated alanine aminotransferase (SGPT), and sorbitol dehydrogenase at both dose levels in
both sexes substantiate other indications of a liver response. Liver pigment analyses
indicated possible hemolysis, cholestasis, and tissue damage and repair: these findings were
more prominent in males than in females. Extramedullary hematopoiesis (especially in spleen)
was consistent with hemolysis. No unique lung toxicity appeared to be related to lung tumors.
This study should be considered along with the related low dose study, 044:085117 (IRDC study
no. 399-063: completed 9/28/89). Acceptable in conjunction with study in Vol. 44. Aldous,
11/16/90.
246-045 085118 Preliminary pathology report for 048:090438, above. This is preceded by a
letter from R. Cardona, who provided evidences that the above study had employed excessively
high dose levels, and thus should not be used for risk assessment. (No CDFA worksheet).
Aldous, 11/2/90.
246-033 070816 12--month interim report for 048:090438, above.
**246-044 085117 Goldenthal, E. I., "Two year oncogenicity study in mice", IRDC Study 399-
063, 9/28/89. CD-1 mice were supplied UDMH (major metabolite of Daminozide) in pH-buffered
drinking water at 0, 1, 5, and 10 ppm (males) or 20 ppm (females). Ninety/sex/group were
initiated on study. Of these, 20/sex/group were sacrificed at 8 months and an additional
20/sex/group at 12 months: the remainder were maintained for 2 years. Purity of UDMH was not
specified. A possible adverse effect was noted: increased incidence of alveolar bronchiolar
adenomas and carcinomas in 20 ppm females. NOEL = 1 ppm in males (pigment accumulation in
liver) and females (single cell necrosis in liver). It appeared that survival was reduced in
20 ppm females during months 19-21: this was the main indication that the MTD may have been
exceeded. There were no (non-neoplasia) treatment effects in lungs corresponding to observed
tumors. Liver findings at 10 or 20 ppm included hypertrophy, hyperplasia, and pigmentation:
none of these changes were marked in degree or incidence. Acceptable (study should be
considered in light of the higher dose study, 048:090438). Aldous, 11/16/90.
CDPR MEDICAL TOXICOLOGY DAMINOZIDE, SUMMARY OF TOXICOLOGY DATA T910801
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246-038 072545 Toth, B., "Hydrazine, methylhydrazine and methylhydrazine sulfate
carcinogenesis in Swiss mice. Failure of ammonium hydroxide to interfere in the development
of tumors." Int. J. Cancer:9:109-118 (1972). C3H mice were given solutions of above
materials in drinking water over their entire lifetimes. Dose levels were 0.001% for
hydrazine, 0.01% for methylhydrazine and 0.01% for methylhydrazine sulfate. Author indicated
that hydrazine and methylhydrazine sulfate as administered increased incidence of lung tumors
(adenomas and adenocarcinomas), and that latency of such tumors was shortened for
methylhydrazine (which markedly increased mortality at the 0.01% level administered). Study
is unacceptable (very limited protocol, only one dose level, insufficient information for
independent assessment), but provides limited information related to neoplasia potential of
UDMH. Aldous (no CDFA worksheet), 11/1/90.
CDPR MEDICAL TOXICOLOGY DAMINOZIDE, SUMMARY OF TOXICOLOGY DATA T910801
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246-038 072548 Toth, B., "The large bowel carcinogenic effects of hydrazines and related
compounds occurring in nature and in the environment". Cancer 40:2427-2431 (1977). A 0.1%
level of 1,1-dimethylhydrazine [= UDMH] in drinking water administered over the lifetime to
Syrian golden hamsters was stated to have reduced survival and to have elicited tumors of the
cecum (including polypoid adenomas and adenocarcinomas). Study is unacceptable (very limited
protocol, only one dose level, insufficient information for independent assessment), but
provides limited information related to neoplasia potential of UDMH. Aldous (no CDFA
worksheet), 11/1/90.
246-038 072764 Toth, B. and Wilson, R.B., "Blood vessel tumorigenesis by 1,2-
dimethylhydrazine dihydrochloride (symmetrical)". Amer. J. Pathol. 64:585-600 (1971).
Administration of 0.001% 1,2-dimethylhydrazine dihydrochloride (symmetrical) in drinking water
of randomly bred Swiss mice reduced survival. There was a remarkable increase in incidence of
angiosarcomas in both sexes, for which background incidence was very low, with apparent
reduction of latency. There was also an increase in comparatively common lung tumors
(primarily adenomas), with a corresponding decrease in latency. Study is unacceptable (test
article is not the isomer of concern, very limited protocol, only one dose level, insufficient
information for independent assessment), but provides limited information related to neoplasia
potential of the related compound, 1,1-dimethylhydrazine (UDMH). Aldous (no CDFA worksheet),
11/2/90.
246-005, 023503: "Carcinogenicity of Hydrazine and 1,1-Dimethylhydrazine for Mouse Lung,"
(l967, publ. in Nature, from Chester Beatty). 1,1-dimethylhydrazine (UDMH, daminozide
contaminant and metabolite), 0.5 mg/mouse, presumed equivalent to about 20 mg/kg, by gavage to
female Swiss mice 5 days a week for 40 weeks. Dose >>>MTD, based on mortality (meaning??);
ONCOGENICITY - Significant increase in lung tumors, alveolo/bronchiogenic adenomas or
adenocarcinomas, at multiple sites. Study UNACCEPTABLE & CANNOT BE UPGRADED for regulatory
purposes, but sheds some light on the tumorigenic properties of daminozide contaminated with
UDMH reported by Eppley. Gee 5/6/85 with second opinion review by Martz 5/14/86.
CDPR MEDICAL TOXICOLOGY DAMINOZIDE, SUMMARY OF TOXICOLOGY DATA T910801
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006, 000939: "1,1-Dimethyl Hydrazine Study Carried Out at Eppley Cancer Institute" 1973
publication; 1,1-Dimethylhydrazine (UDMH -no purity information), 0.01% in drinking water
until 100% mortality; Report consists of a second opinion diagnosis of hepatocellular tumors
by pathologist Dr. Stan D. Vesselinovitch (see letter dated 4-10-84 in same volume) of slides
previously diagnosed by principal investigator Bela Toth. Many liver vascular tumors were
rediagnosed as being non-neoplastic hepatotoxic lesions. Discrepancy is moot because new
studies with daminozide and UDMH are in progress, see -018 above. Martz 7/15/87.
REPRODUCTION, RAT
** 023, 059837: "Two Generation Reproduction Study with ALAR in Rats (One Litter per
Generation)" (Hazleton, HLA Study No. 6111-102, 5/26/87) Alar (Lot No. MO 55030, 99% pure
daminozide) fed to 25 Sprague Dawley rats/sex/dose at 0, 100, 1000 or 10,000 ppm in diet in F
0
and F generations; F and F generations exposed in utero and via lactation; study terminated
1 1 2
when F reached 21 days of age; NO ADVERSE EFFECT; initially reviewed as not upgradeable with
2
insufficient dose selection rationale - no evidence of toxicity (in any parameter measured) in
F and F rats fed the highest dose. NOEL > 10,000 ppm (HDT). Becker 10/28/87, Kishiyama &
0 1
Davis 10/11/88. Document 246-047, Record 088035, contains a rebuttal and a review by FAO/WHO
panel accepting the study with a NOAEL = 1000 ppm. Re-consideration of the total data base
and the use of a high dose of 10,000 ppm, the study is ACCEPTABLE. Gee, 3/16/90.
246-037 072998 Partial duplicate of 023:059837, above.
030, 066544: "Data Evaluation Record. Alar. Two-Generation Reproduction Study in Rats"
Contract review by Dynamac Corporation for EPA. EPA set parental toxicity NOEL = 1,000 ppm &
LEL = 10,000 ppm based on reduced body weight in F1 parental males; reproductive/developmental
toxicity NOEL = 100 ppm & LEL = 1,000 ppm based on delayed mating in treated males. EPA
accepted the study as Core-Minimum. CDFA review did not alter previous review. Kishiyama &
Davis 10/11/88.
CDPR MEDICAL TOXICOLOGY DAMINOZIDE, SUMMARY OF TOXICOLOGY DATA T910801
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013, 036933: "Chronic (2-Year) Feeding Studies With B-995 in Rats and Dogs -Reproduction and
Lactation," FDRL, 11/15/66; daminozide (no purity information) fed to 20 rats/sex at 0 or 300
ppm in F generation and to 10/sex/group for F and F parental generations, F part of
0 1 2 0
chronic combined study (record #036932), NO ADVERSE EFFECT reported, INCOMPLETE (missing
individual data), UNACCEPTABLE, CANNOT BE UPGRADED (too few animals, single dose (300 ppm)
with no justification, no analysis of diet). Gee 11/26/85.
TERATOLOGY, RAT
** 246-004, 000950: "Teratologic Evaluation of Alar Technical in Sprague-Dawley Rats" FDRL,
3/6/79; daminozide (Alar technical, no purity information) at 0, 85, 390 and 1800 mg/kg by
oral gavage days 6-15 of gestation; Positive control was aspirin at 1800 mg/kg; NO SIGNIFICANT
ADVERSE EFFECTS in absence of maternal toxicity; NOEL = 390 mg/kg for developmental and
maternal effects. ACCEPTABLE. Gee 5/3/85 with second opinion by Parker on 3/26/86.
246-014, 036937: "Teratogenic Study with ALAR in Albino Rats," IBT, 11/28/72; Invalid IBT
study, IBT No. B1708, Dose levels were 250, 500 mg/kg; days 6-15 to rats. Gee 5-3-85
246-005, 023505: "Teratologic Assessment of Maleic Hydrazide and Daminozide, and Formulations
of Ethoxyquin, Thiabendazole and Naled in Rats," [J. Env. Sci. Health B14(6), 563-577, 1979],
Daminozide (99%) at 0, 200, 600 or 1000 mg/kg days 6-15 of gestation by oral gavage; NO
ADVERSE EFFECT reported; INCOMPLETE (No individual data, no analysis of dosing solution);
UNACCEPTABLE (need justification of dose-dose levels not high enough, justification &
description of use of water as a vehicle) Could be upgraded. Gee 5/6/85.
TERATOLOGY, RABBIT
** 246-013, 036935: "Teratology Study in Rabbits (Alar)" IRDC, 8/7/85; daminozide, 99% pure,
by oral gavage in CMC at 0, 50, 150 or 300 mg/kg/day to 16 rabbits/ group, days 7-19 of
CDPR MEDICAL TOXICOLOGY DAMINOZIDE, SUMMARY OF TOXICOLOGY DATA T910801
Page 17
gestation; maternal effects were 1 death and reduced weight gain at 300 mg/kg; NO ADVERSE
DEVELOPMENTAL EFFECTS, developmental NOEL > 300 mg/kg. ACCEPTABLE with minor variances. Gee
11/26/85.
246-007, 000957: "Toxicological Studies on Succinic Acid 2,2-dimethyl Hydrazide (ALAR)--
Somer’s Test"; Brown Biological Laboratories, no date; daminozide (no purity information) at
0, 42 or 126 mg/kg by oral gavage days 7-16 of gestation, half of animals sacrificed day 28,
remainder allowed to go to term; INSUFFICIENT INFORMATION FOR ADVERSE EFFECTS ASSESSMENT;
INCOMPLETE (missing some individual data). UNACCEPTABLE AND CANNOT BE UPGRADED (no toxicity
at high dose). Gee 5/3/85.
GENE MUTATION
007, 000951: "Mutagenicity Evaluation of B995 (ALAR)" Litton Bionetics, 5/18/77; Ames test
with Salmonella strains TA1535, TA1537, TA1538, TA98, and TA100 and Σαχχηαροµψχεσ χερεϖισιαε
strain D4; Daminozide (no purity information) at 0.1, 1, 10, 100, or 500 ug/plate with and
without activation, No effects reported, but has insufficient information for assessment;
UNACCEPTABLE, CANNOT BE UPGRADED (too few plates, dose level too low, and no repeat test for
confirmation). Gee 5/3/85
006, 000935: "P7642: Investigation of Mutagenic Activity in the TK+/- Mouse Lymphoma Cell
Mutation Assay;" Life Sci. Res., 9/82); Mouse lymphoma L5178Y cells; Daminozide technical (no
purity information) at 0, 1500, 2000, 2333.3, 2666.7 or 3000 ug/l with and without activation;
NO ADVERSE EFFECT indicated; originally accepted on 5/7/85 (J.R. Gee), but downgraded to
UNACCEPTABLE on basis of no repeat trial. Gee 5/7/85 and 7/2/87.
006, 000943: "Uniroyal Summary and Review of Daminozide and UDMH Oncogenicity Studies"--
Mutagenicity Studies, no date; Brief literature review of daminozide and UDMH mutagenicity
studies, Cites ADVERSE EFFECT in a mouse lymphoma assay for UDMH and states that UDMH was
CDPR MEDICAL TOXICOLOGY DAMINOZIDE, SUMMARY OF TOXICOLOGY DATA T910801
Page 18
inactive or marginally active in several Ames (Salmonella) assays, Very Brief Summary,
SUPPLEMENTAL.
** 246-051 097254 "Salmonella/Mammalian-Microsome Plate Incorporation Mutagenicity Assay
(Ames Test) with a Confirmatory Assay", (Richard H.C. San, Ph.D. and J. Blair Shelton, B.S.,
Microbiological Associates, Inc., 9900 Blackwell Road, Rockville, MD, Report # T9292.501014,
3/21/91). Daminozide, lot #806M006NB, 99.8% purity, was tested in the reversion assay with
Salmonella strains TA98, TA100, TA1535, TA1537 and TA1538 with and without metabolic
activation by Aroclor 1254 stimulated rat liver S-9 fraction with 3 plates/strain/dose in 2
experiments at concentrations of 0 (DMSO control), 667, 1000, 3333, 6667 and 10000 ug/plate.
No adverse effects were noted (no increase in the number of revertant colonies). Acceptable.
(Green, Kellner and Gee, 7/24/91).
246-050 096353 Partial duplicate of 051:097254, above.
CHROMOSOME EFFECTS
246-007, 000953: "Dominant Lethal Assay of Alar in the Male Mouse" Huntingdon Res. Ctr.,
6/22/73; daminozide (no purity information) at 0, 10, 300 or 10,000 ppm to 20 CFLP mice/group
(3 doses based on preliminary study); MTD not achieved; NO ADVERSE EFFECT indicated;
INCOMPLETE (no individual data); UNACCEPTABLE, CANNOT BE UPGRADED (no concurrent positive
control, too few females, only four weeks of testing). Gee 5/3/85.
CDPR MEDICAL TOXICOLOGY DAMINOZIDE, SUMMARY OF TOXICOLOGY DATA T910801
Page 19
** 246-051 097253, "Chromosome Aberrations in Chinese Hamster Ovary (CHO) Cells", (Donald L.
Putman, Ph.D. and Marcia J. Morris, B.A., Microbiological Associates, Inc., 9900 Blackwell
Road, Rockville, MD., Report # T9292.337, 2/20/91). Daminozide, lot #806M006 NB, 99.8%
purity, was tested for chromosomal aberrations in Chinese Hamster ovary cells (CHO-K ) with
1
and without metabolic activation by Aroclor 1254 stimulated rat liver S-9 fraction with 2
cultures/dose at levels of 0 (DMSO), 250, 500, 1000, and 2000 ug/ml. No adverse effects were
noted (no increase in the proportion of aberrant metaphases). Acceptable. (Green, Kellner
and Gee, 7/25/91).
246-050 096354 partial duplicate of 246-051:097253, above.
DNA DAMAGE
** 246-006, 000934: "P7642: Assessment of Its Ability to Induce Primary DNA Damage in Strains
of Εσχηεριχηια χολι" Life Sci. Res., 7/16/82, Ε. χολι with and without activation; daminozide
(no purity information) at 250, 1000, 2500 and 10,000 ug/ml; NO EVIDENCE OF DNA DAMAGE;
COMPLETE, ACCEPTABLE. Gee 5/7/85.
** 246-006, 000936: "P7642: Assessment of its Ability to Induce Genetic Damage in
Σαχχηαροµψχεσ χερεϖισιαε" Life Sci. Res., 1/21/83, Σαχχηαροµψχεσ χερεϖισιαε strain D6;
Daminozide (no purity information) at 1-2000 ug/ml (11 concentrations) with and without
activation; NO EVIDENCE OF ADVERSE EFFECT (mitotic crossing over); COMPLETE, ACCEPTABLE. Gee
5/6/85.
NEUROTOXICITY
Not required at this time.
GENOTOXICITY STUDIES WITH UDMH OR RELATED HYDRAZINES
CDPR MEDICAL TOXICOLOGY DAMINOZIDE, SUMMARY OF TOXICOLOGY DATA T910801
Page 20
GENE MUTATION
019 057778 "Ames/SALMONELLA Plate Incorporation Assay" (Pharmakon Research International,
PH 301-UN-005-086, 12/9/86) Unsymmetrical dimethyl hydrazine (UDMH), lot JS-34, E732-086, no
purity stated; tested with Salmonella strains TA1535, TA1537, TA1538, TA98 and TA100 at 0, 25,
83.3, 250, 833, 2500 and 5000 ug/plate in triplicate, one trial with cytotoxicity (reduced
colonies or change in background lawn) at 5000 ug/plate; NO EVIDENCE FOR AN INCREASE IN
REVERSION RATE; conducted in accordance with EPA guidelines as modified in May, 1987.
SUPPLEMENTAL DATA (derivative of daminozide). Gee 12/23/87.
019 057777 "CHO/HPRT: Mammalian Cell Forward Gene Mutation Assay" (Pharmakon Research
International, 1/16/87, PH 314-UN-001-086) Unsymmetrical dimethyl hydrazine (UDMH), lot JS-
34, E732-086), purity not stated; tested with Chinese hamster ovary cells for gene mutation of
the HGPRT locus to thioguanine resistance at 0, 50, 100, 250, 500, 750 or 1000 ug/ml with no
cytotoxicity, the high concentration limited by the solvent (HCl); no consistent effect on
mutation frequency although some samples were statistically significant - RESULTS CONSIDERED
EQUIVOCAL; SUPPLEMENTAL DATA (derivative of daminozide). Gee 12/23/87.
CDPR MEDICAL TOXICOLOGY DAMINOZIDE, SUMMARY OF TOXICOLOGY DATA T910801
Page 21
034 070968 "Unsymmetrical Dimethylhydrazide (UDMH) CHO/HPRT Mammalian Cell Forward Gene
Mutation Assay" (Pharmakon Research International, Inc., Laboratory Project ID: PH 314-UN-
001-88, 9/12/88) Duplicate cultures of Chinese Hamster Ovary cells (CHO-K1-BH4) were exposed
to UDMH = 1,1-Dimethylhydrazine at 0, 50, 100, 250, 500, 1000, 2500, 3750, and 5000 ug/ml + S9
in one assay and at 0, 50, 100, 250, 500, 1000, 1500, 2000, 2500, 3000, 3500, 3750, 4000,
4500, and 5000 ug/ml + S9 in a second assay; NO ADVERSE EFFECT-A 3-fold increase in mutation
rate in the first assay at 3750 ug/ml + S9 was not confirmed in the second assay; cytotoxicity
was demonstrated; SUPPLEMENTAL STUDY; Davis 10/26/88.
CHROMOSOME EFFECTS
019 057775 "In vitro Chromosome Aberration Analysis in Chinese Hamster Ovary (CHO) Cells"
(Pharmakon Research International, 12/6/86, PH 320-UN-002-86), UDMH (25,000 ppm in stock - no
solvent given); tested in CHO cells stated in the report to be 0, 500, 1500 or 5000 ug/ml, but
from raw data, high concentration is 500 ug/ml, 5-hour exposure (6 hours according to raw
data) with and without Aroclor-induced rat liver followed by 18 hours in BrdUrd; duplicate
cultures, scored 2 x 50 cells per concentration; solvent was 0.25 N HCl at 1 or 2% final
concentration (unclear which from report); NO INCREASE IN ABERRATIONS reported; SUPPLEMENTAL
DATA (derivative of daminozide). Gee 12/22/87. EPA: UNACCEPTABLE. See Pesticide and Toxic
Chemical News, June 17, 1987.
DNA DAMAGE
019 057776 "Rat Hepatocyte Primary Culture/DNA Repair Test" (Pharmakon Research
International, Inc., PH 311-UN-0001-86, 12/6/86) Unsymmetrical dimethyl hydrazine (UDMH),
lot JS-34, E732-086, 25 mg/ml; tested with rat hepatocytes at 0, 8.3, 25, 83 and 250 ug/ml, 18
- 20 hours; scored 150 cells by autoradiography per concentration; 0.25 N HCl, the vehicle; NO
INCREASE IN UNSCHEDULED DNA SYNTHESIS reported; SUPPLEMENTAL DATA (derivative of daminozide).
Gee 12/23/87.
