NITROCELLULOSE
Health Advisory
Office of Drinking Water
U.S. Environmental Protection Agency
Washington, DC 20460

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PREFACE
This report was prepared in accordance vich che Memorandum of Understa-dirg
becveen che Department of che Army, Deputy for Environment, Safety and
Occupational Healch (OASAd&L)) and the U.S. Environmental Protection Agency
(EPA), Office of Drinking Water (ODW), Criteria and Standards Division, for the
purpose of developing drinking water Health Advisories (HAs) for selected
environmental contaminants, as requested by the Army.
Health Advisories provide specific advice on the levels of contaminancs in
drinking water at which adverse health effects would not be anticipated and which
include a margin of safety so as to protect the mosc sensitive member of the
population at risk. A Health Advisory provides health effects guidelines and
analytical methods and recommends treatment techniques on a case-by-case basis.
These advisories are normally prepared for One-day, 10-day, Longer-term and
Lifetime exposure periods where available toxlcological data permit. These
advisories do not condone the presence of contaminants in drinking water; nor are
they legally enforceable standards. They are not Issued as official regulations,
and they may or may not lead to the Issuance of national standards or Maximum
Contaminant Levels (MCLs).
This report Is the produce of the foregoing process. Available toxlcological
data, as provided by the Army, on the munitions chemical, nitrocellulose (NC),
have been reviewed, and the relevane findings are presented In this report in a
manner so as to allow for an evaluation of the data without continued reference to
che primary documents.
Significant chemical and physical properties of NC as well as potential
sources of exposure are provided as a means of identifying the subject chemical.
Data on the pharmacokinetic properties, although somewhat limited in scope, are
summarized. All available toxlcological data, including short-term, longer-term
and lifetime feeding studies in three species* as well as the results of
reproductive and genetic toxicology studies, have been reviewed, and those effects
deemed relevant to Che ingestion of NC are suaaarlzed. Results of Immunologic and
carcinogenic studies ars also Included.
This roport also includes a section describing state-of-the-art methods of
analyses foe BC In drinking water and Includes essential treatment techniques
appropriate for NC removal froa an affected water supply, should the levels of NC
in the drinking water reach a level considered unpalatable due to taste, clarity
or similar indicators.
This report has been submitted to a critical review by the EPA to include a
panel of Health Effects Branch (HEB) toxlcologlsts. Their comments, as
appropriate, have been Incorporated Into this report.
A companion document, "Data Deficiencies/Problem Areas and Recommendations

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for Additional Data Base Development for Nitrocellulose" ls included in ;-13
report under Appendix 2.
I would like to express ny thanks to Dr. John Glennon, Life Systems, Inc.,
who afforded valuable coordination and logistical assistance. I also thank
Dr. Janet Normandy, Ms. Lori Gordon and Dr. William Hartley who provided the
extensive technical skills required for the preparation of this report.
I am grateful to the following members of the EPA Tox-Review Panel,
Dr. Kenneth Bailey, Dr. Aablka Bathija, Dr. Bill Marcus and Mr. Bruce Mintz, who
took time to review this report and Co provide their invaluable input.
Finally, I would like to thank Dr. Edward Ohanlan, Chief, Health Effects
Branch, Dr. Joseph Cotruvo, Director Criteria and Standards Division, and Dr.
Penelope Fenner-Crlsp, Manager, Health Advisory Program, for providing me vich the
opportunity and encouragement to be a part of this project.
The preparation of this Advisory was funded in part by Interagency Agreement
(IAG) 85PP5869 between the U.S. EPA and the U.S. Army Medical Research and
Development Command (USAMKDC). This IAG was conducted with the technical support
of the U.S. Army Biomedical Research and Development Laboratory (USABRDL).
Krishan Khanna, Project Officer
Office of Drinking Water

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TA3L- Of CONTENTS
°3Ee
I.	INTRODUCTION		I-1
II.	GENERAL INFORMATION		II-l
III.	SOURCES OF EXPOSURE		Ill-1
IV.	PHARMACOKINETICS		IV-1
V.	HEALTH EFFECTS		v-l
A.	Short-Term Exposure	'		V-l
1. Primary Skin and Eye Irritation		V-l
B.	Longer-Term Exposure		V-l
1.	Thirteen-Week Studies		V-l
2.	Lifetime Exposure		V-3
C.	Genetic Toxicology		V-7
D.	Carcinogenicity		V-8
E.	Reproductive Effects		V-9
F.	Teratogenicity		 . . 		V-10
G.	Immunologic Effects		V-10
VI.	HEALTH ADVISORY DEVELOPMENT		VI-1
VII.	ANALYSIS		VII-l
VIII.	TREATMENT	VIII-1
IX.	CONCLUSIONS AND RECOMMENDATIONS		IX-1
X.	REFERENCES	'		X-l
LIST OF TABLES
Table
Il-i General Chemical and Physical Properties of Cellulose 		II-2
Trinitrate

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APPENDiC-S
Appendix
A1	Calculation Mechods	 AL-l
A2	Daca Deficiencies/Problem Areas and Recommendations for
Additional Data Base Development for Nitrocellulose	 a2-1

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I. INTRODUCTION
Th« Health Advisory (HA) Program, sponsored by the Office of Drir.kir.g
Water (ODW), provides information on the health effects, analytical
methodology and treatment technology that would be useful in dealing vie11 che
contamination of.drinking water. Health Advisories describe nonregulatorv
concentrations of drinking water contaminants at which adverse health effects
would not b« anticipated to occur over specific exposure durations. Health
Advisories contain a margin of safety to protect sensitive members of the
population.
Health Advisories serve as Informal technical guidance to assist Federal,
State and local officials responsible for protecting public health when
emergency spills or contamination situations occur. They are not to be
construed as legally enforceable Federal standards. The Advisories are
subject to change as new information becomes available.
Health Advisories are developed for One-day, Ten-day. Longer-term
(approximately 7 years, or 10Z of an individual's lifetime) apd Lifetime
exposures based on data describing noncarcinogenlc end points of toxicity.
Health Advisories do not quantitatively incorporate any potential carcinogenic
risk from such exposure. For those substances that are known or probable
human carcinogens, according to the Agency classification scheme (Croup A or
B), Lifetime HAs are not recommended. The chemical concentration values for
Croup A or B carelnogens are correlated with carcinogenic risk estimates by
employing a cancer potency (unle risk) value together vlth assumptions for
lifetime exposure and the consumption of drinking water. The cancer unit risk
is usually derived from the linear multistage model with 95 J upper confidence
limits. This provides a low-dose estimate of cancer risk to humans that is
considered unlikely to pose a carcinogenic risk In excess of the stated
values. Excess cancer risk estimates may also be calculated using the
One-hit, Weibull, Logic and Problt models. There is no current understanding
of the biological mechanisms involved in cancer to suggest that any one of
these models is able to predict risk more accurately than another. Because
e.ach model la based upon differing assumptions^ the estimates that are derived
can differ by several orders of magnitude.
I-i

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CZSI3AL INFORMATION
As described by Sullivan ec al. (1978), nitrocellulose, or cellulose
trinitrate. is a non-volatile, fibrous, cotton-like, white solid (specific graviv
1.66) consisting of chains of beta, 1 co 4 linked glucoside units in which the
hydroxy1 groups of the glucose subunits have reacted to form nitrate esters. The
chemical formula is approximately	(ONO^) ^ lfl» c^e °olecular weight depends
on chain length, and the degree of polymerization (DP) varies appreciably. Cotton
lmters and wood pulp used for the preparation of military grades of NC have a DP
of approximately 1,000 to 1,500. The fully nitrated form of NC has a nitrogen
concent of 14.14Z and a formula weight of 297.14. This degree of nitration is
difficult to accomplish (Department of the Army Technical Manual TM9-1300-214,
1967). Nitrocellulose is extremely flammable and has a flash point-of 12.8°C.
The melting point range Is 160* to 170'C — also its autolgnltlon temperature.
"Guncotton", military grade cellulose nitrate, contains 13.5Z nitrogen and is
the most highly nitrated form. Theoretically, mononitrated, dinitrated, and
trinitrated cellulose contain 6.8Z N, 11.1Z N, and 14.1Z N, respectively.
"Guncotton" is essentially fully nitrated and, therefore, can be considered to be
a crude cellulose trinitrate contaminated by traces of less completely nitrated
esters (Sullivan et al., 1978). The general properties of cellulose trinitrate
are susmarized in Table II-l.
The nitrate esters, in general, are soluble in esters, aldehydes, and
ketones. The more completely nitrated a cellulose is, however, the smaller the
range of materials in which It is soluble. The less nitrated forms of cellulose
nitrate are also very soluble In methanol, benzene, toluene, and mixtures of ether
and ethanol. These compounds (pyroxylin: 8Z to 12Z N and pyrocellulose: 12.5% N)
are forms of collodion and are used chiefly as lacquers, ink bases, and filter
membranes, and in veterinary medicine for wound closure. Cellulose trinitrate is
insoluble in water, ethanol, ethyl ether, and benzene but completely miscible in
acetone, methylethyl ketone, tetrahydrofuran, as well as nitrobenzene and ethyl-,
butyl-, and amylacetstes (Sullivan et al., 1978).
Nitrocellulose Is resistant to biological degradation and is a persistent
compound in ths environment. It has been suggested by Sullivan et al. (1978) that
cellulose trlscststs and cellulose trinitrate are reslstane to enzymatic attack.
These authors further ststed that although direct blodegradatlon of nitrocellulose
does not tees to bo feasible, alkaline hydrolysis yields material that can be
decomposed by microbial activity. Nitrocellulose, therefore, .an be characterized
as being persistent la the environment unless chemically altered.
Based on aa extensive volume of literature dealing with the adsorptlve
capacity of nitrocellulose for biologically Important macromolecules, Sullivan et
al. (1978) have Indicated that nitrocellulose strongly adsorbs and concentrates
DNA and various RNA molecules from solution, and adsorbs proteins, and polypeptides
under-various conditions. Proteins with an uncoiled structure or molecular weight
greater than 10 are strongly bound. Denatured proteins are also strongly
adsorbed, especially under acidic conditions.
II-1

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TABLE II-1 GENERAL CHEMICAL AND PHYSICAL PRCPERTIES
OF CELLULOSE TRINITRATE3
CAS Number
Names
Molecular velghc
Empirical formula
Structure
9004-70-0
Cellulose trinitrate; nitrocellulose;
guncotton
Varies with chain length
[C6H7°2(°N02)3]n
mjCONO,	HjCONO,	N,C0*0,
H	ONO.	m	OMO,	H	CO,
Color
Physical state
Specific gravity
Percent nitrogen
carbon
oxygen
hydrogen
Solubility characteristics
Melting point.
Autoignitlott teaperatux*
Flash polnfc
Whitish
Fibrous solid
b/
1.66
14.1
24.3
59.3
2.4
Iosoluabls in water* ethanol, benzene and most
other solvents. Soluble in 25 parts of
1:3 alcohol:ether; soluble in acetone in
all proporstions; soluble in methylethyl
ketone, nitrobenzene, tetrahydVofuran, and
ethyl, butyl and aaylacetate.
160-170* C
160-170* C
12.7* C
References: Sullivan et si., 1978; Havley, 1981; RTECS, 1986.
Approximate, depends on processing and chain length.
11-2

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III. SOURCES OF EXPOSURE
Nitrocellulose (NC) is a principal ingredient of propellants, smokeless
powder, rockee fuel, ball powder, mortar increments, and some explosives
(Rosenblatt et al., 1973). It is produced for military use at selected Arav
ammunition plants (AAP) by treating cotton linters or wood pulp with mixed nitric
and sulfuric acids at 308C. The resulting slurry is centrifuged to remove cost of
the acid, treated with several changes of boiling water, washed with a heavy
stream of water, and finally screened to remove most of the water (Department of
the Army Technical Manual TM9-1300-214, 1967). Production requires 16 to 22
gallons of process water per pound of NC produced. Most of this water is
discharged and contains, la addition to NC, 0.7 to 1.0 pound of sulfuric acid ar.d
0.3 to 0.4 pound of nitric acid per pound of NC produced, thus resulting in a low
pH (Rosenblatt et al., 1973; Helton, 1976). Nitrocellulose fines are found in
production wastewater because of settling pit overflow and some escape after
flowing through the waste acid neutralization process lines (Huff et al., 1975).
Helton (1976) analyzed samples of NC fines from wastewaters of an AAP and found
that more than 99Z of the particulate material was military grade NC with an
average nitrogen content of 12.9Z and particle size greater than 5 microns.
Sullivan et al. (1978) Indicated that these data suggested that suspended solids
below 0.8 microns contained significant portions of nonnltrated cellulose and
other materials while those greater than 44 microns were mostly NC.
III-l

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IV. PHAJ^-ACOKINETICS
There art currently no available daca co suggest chat nitrocellulose ca-i be
absorbed from any route. A study in rats has shown that NC appears to reaain
within the gastrointestinal tract until it is excreted in the feces. Ccher
available data are inconclusive.
^ Ellis ec al. (1976) studied the absorption, distribution, and excretion o£
C-nitrocellulose (prepared froo cotton grown in the presence of D-glucose-t'L- *C
and nitrated by standard procedures) in two male Charles River CD rats. Each rat
was fasted overnight before being given NC orally either in an aqueous suspension
or in a suspension of 0.2Z methyl cellulose-O.4Z Tween 80 (MC-TW80) at a volume cf
1 ml/100 gm (presumably bodyveight; about 20,000 dpm/ml). After dosing, each rat
was placed Immediately In a "Roth-Delmar" metabolism chamber. Expired CO , feces,
and urine vera each collected separately In the apparatus. To ensure that
sufficient radioactivity was administered, the dosing was repeated dally for 4
days. Twenty-four hours after the last dose, the animals were anesthetized and
aortic blood was collected. Liver, spleen, brain, kidneys* lungs and thigh muscle
along with the stomach, small Intestine, cecum, large intestine and their contents
were removed and each assayed for radioactivity via liquid scintillation
spectrometry.
After repeaeed oral doses of radiolabeled nitrocellulose, no detectable
radioactivity was found in any tissue or body fluid. Radioactivity was recovered
only in the various components of the gastrointestinal (GI) tract plus contents
and in the feces (percentage dpm recovered not specified). From these results,
the authors concluded that the nitrocellulose molecule was not absorbed by the
rat.
Ellis et al. (1980) conducted a mass-balance metabolism study of
nitrocellulose using a dog fed 90 g (mg/kg dose could noe be determined from data
provided) of wet nitrocellulose (27.9 g dry weight). After 24, 48, 72 and 96
hours, the feces wera collected and analyzed by the method of Sellg (1961) which
involves reduction of nitro groups and titration to a colorlmetrlc endpolnt.
During the fIrst~48-hour period, 9.5 g (dry weight) of NC were recovered
representing a recovery of 34Z (g recovered/g fed, dry weight). No further
nitrocellulose vu Identified In later fecal samples. U« can draw no useful
generalizations from these results since they could indlcaee either that the
method of analysis vu not particularly effective, that soma nitrocellulose may
have been abaorbad from Che dlgeatlve trace of this dog, or that NC was denitrated
by gut bactarla.
IV-1

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Currently available studies on the health effects of nitrocellulose suggest
that it is not "toxic" unless ingested in enoraous quantities such as 10" of c-e
total diet. In those cases, death from intestinal impaction may occur just as lc
does in animals fed the same dietary levels of non-nitrated cotton linters.
A. Short-Term Exposure
The acute oral toxicity of nitrocellulose is very low in mice and rats as
evidenced by LD 's of more than 5,000 mg/kg in both species. No acute toxicity
data of nitrocellulose are provided in the 1986 edition of the Registry of Toxic
Substances (RTECS, 1986).
Lee et al. (I97S) used male and female Charles River rats and male and female
albino Swiss mice (apparently 10/sex/dose; number and range of doses noe clearly
specified) to determine the acute oral LD of nitrocellulose. Each dose of
nitrocellulose, suspended in water ac a final concentration of SZ NC (dry weight
basis), was divided in half and given 30 minutes apart due to the large volume
necessary to administer a dose of 5,000 mg/kg (the authors did not indicate the
volume administered). Test animals were fasted for at least 16 hours prior to
oral dosing via gastric lntubaelon. After treatmene, the survivors were observed
daily for 14 days for delayed mortality or signs of toxicity. The LD,_ was
calculated by a computer program based on the method of maximum likelihood of
Finney (1971). The LD^'s for nitrocellulose in male and female racs and in male
and female mice were greater than 5,000 mg/kg. No toxic signs were observed in
animals receiving the highest dose of nitrocellulose. Two of 10 male mice given
5,000 mg/kg died without any apparent gross lesions. No other animals died at any
dose level.
I. Primary Skin and Eye Irritation
Using the modified Drslze method for skin and eye irritation, a 33Z
concentration of NC in vater was applied directly to the eye or shaved skin of the
rabbit and evaluated ac 24 and 72 hour* (Lee et al., 1975). This concentration of
NC was noc Irritating to either rabbit skin or eye.
B. Longer-Term Exposure
1. Thirteen-Week Studies
No advert* effaces that cam be related to ths chemical characteristics of
nitrocellulose wars Identified in the available thirteen-week studies In dogs,
rats, and mice (Ellis et al., 1976). Intestinal blockage was the cause of death
in high-dose mice (10Z NC) as wall as la control mice fed the same amount of
cotton linters (10Z of total diet).

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•His ec al. ( 1976) conducted thirteen-week studies of the oral tc~.i:.:
nitrocellulose using dogs, rats and mice. In the studies with dogs, 20 vour.g
healthy beagle dogs (7.2 to 13.6 kg) were divided into five groups of two a'aies
and two females.. Three groups were administered NC as IX, 32, or 102 of their
feed as dry weight. (Table Al-1, Appendix Al, describes method used to calculate
doses. Using the authors' data for body weight at week 4 and average intake data
for weeks 1 through 4, these doses were calculated to correspond to approximate iy
518, 1,900 and 6,890 og/kg/day, respectively, for males and 610, 2,976 and 8,485
rag/kg/day, respectively, for females.) The fourth group received a mixture of 10"
(approximately 6,181 and 8,627 mg/kg/day, males and females, respectively,
calculated as above) of cotton cellulose linters and served as a cotton control to
determine if any observed effects were due to the passage of non-nutritive bulk
through the gastrointestinal trace. The fifth group served as an untreated
control.
The tests included hematology, clinical blood chemistry and bromo-
sulfophthalein (BSP) retention tests. Ac termination, the dogs were sacrificed
and examined for treatment-related gross lesions and organs were weighed. Various
tissues were processed for microscopic examination of lesions.
Feeding NC to dogs at up to 10Z of their diets or cotton linters at 10Z for
13 weeks did not cause any adverse effects. Dogs fed 10Z NC or linters ate about
15Z more than the others* indicating the test materials were merely non-nutritive
bulk. All dogs, including the uncreated and coccon controls, shoved some
variations in body veighc, peripheral blood elemencs and various clinical
chemistry tescs but all data were reported to be within normal limits.
Nitrocellulose was not reported Co cause any ereacmenc-relaced gross or
microscopic lesions.
In thirteen-week studies vich racs, Ellis ee al. (1976) used 40 male and 40
female young healchy CD racs divided Into five groups, each consisting of eight
males and eighe females. Three groups of racs were fed NC ae 1Z, 3Z, or l'OZ in
the feed as dry veighc. The fourch group, che coccon concrols, vas fed cotton
linters as 10Z of che feed. (Ref. Table Al-1: Using che authors' data for the
body weights averaged from weeks 0, 4, 8 and 13 and che average intake over weeks
1 through 13* these dose* correspond to approximately 667 and 2,366 mg/kg/day for
1Z and 3Z» respectively, in che males and 820 and 2,673 mg/kg/day, respecclvely,
in che feaalsa. Measured incakes for che racs fed 10Z NC or 10Z cotton linters in
che diet war* more than double chose of che uncreaced concrol racs. The authors
indicated that at 10Z NC or 10Z coccon llncers in che dlec, enormous mounds of
whlce fluffy material were scattered all around the cage. Therefore, accual
lncake of NC or cotton linters at 10Z in che diet cannot be calculated with any
degree of accuracy.) Ths fifth group* the untreated controls* vas given the
povdered standard rodent chov without NC. Experimental procedures vere similar to
those used with dogs (Ellis et al., 1976) except that BSP retention tests were not
performed.
V-2

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Nitrocellulose arid the cotton linters at 102 in the feed apparent:-/ actec =s
a non-nutritive bulk ingredient which the rats attenpted to remove. These rats
gained less weight than did the untreated control rats. The authors concluded
that this was because they did not absorb enough of the nutritive portion o: the
feed. Rats fed the low or middle levels of NC apparently received enough
nutritional intake and gained weight comparably to the untreated controls.
Nitrocellulose administration did not result In any significant changes in
peripheral blood elements or clinical blood chemistry or any apparent gross or
microscopic lesions. Observed changes in the absolute organ weights of the liver,
kidney and spleen of male rats fed 10Z NC or cotton linters were statistically
significant when compared to untreated controls. These changes were attributed by
the authors to depressed body weight gain, as evidenced by comparable organ
weight/body weight ratios.
In the thirteen-week studies with mice, Ellis et al. (1976) used 40 male and
40 female young healthy albino Swiss mice divided into five groups, each
consisting of eight males and eight females. The treated groups were fed the same
diets as prepared for the rats: 1Z, 3Z, or 10Z of NC as dry weight in feed.
Control mice received either the powdered standard rodent chow alone (as untreated
controls), or 10Z of cotton linters (as cotton controls). (Ref. Table Al-l:
Using the authors' data for the body weights averaged froa weeks 0, 4, 8 and 13,
and the average intake for weeks 1 through 13, these doses correspond to
approximately 1,690 and 5,062 mg/kg/day for 1Z and 3Z, respectively, in males and
1,741 and 7,000 mg/kg/day, respectively, in females. As in the rat study
previously described, the actual Intake of 10Z NC or 10Z cotton linters in the
diet cannot be calculated with any degree of accuracy due to excessive scattering
of the fibers.) The procedures used were similar to those described for rats
except that clinical blood chemistry tests were not performed in mice. Blood
samples for terminal hematology were collected by heart puncture under ether
anesthesia.
Mice fed the low, middle or high levels of NC showed no apparent adverse
effects from the chealcal nature of this compound. In the first few weeks of the
study, there were weight losses and deaths, apparently due to the fibrous nature
of the substance. In mica fad the high level (10Z) of NC as wall as In mice fed
10Z of cotton llntera. The authors concluded that deaths were due to the blocking
of tha lover part of tha gastrointestinal tract by masses of the fibers,
particularly In tha regions where water is removed froa the chyme. Additional
animals fro* a chronic study (saaa shipment and identical levels of NC) were added
to this 13-vaek study to coapensate for tha early losses. No adverse effects or
changes vara obaerved In peripheral blood elements nor were any treatment-related
gross or microscopic lesions reported by the authors.
2. Lifetime Exposure
Long-term studies conductsd in dogs, rata and mica Indicated a dose-related
Increase in total feed consuaptlon and decreases in weight gain in high-dose
V-3

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NC-created rats and mice. Histopathologic evaluations of dcgs, rats a-.i -_^e
created for up to 24 aonths showed naturally occurring (as oppcsed :o
treatment-related) lesions in the thyroid and lungs in dogs, the lur.gs in rats a-i
several organs in mice (kidneys, adrenals, liver and intestines). Adverse effects
causing early deaths due to intestinal impaction occurred in high-dose aice.
Ellis ee al. (1980) studied the effects of NC administered orally at dose
levels up to 10! NC in feed for dogs, rats and mice for 12 and 24 months.
Three groups of beagle dogs, six of each sex per group, were administered NC
at 1Z, 3Z or 10Z In the feed calculated on a dry weight basis for up to 24 months.
(Ref. Table Al-1: Using the authors' data for the average body weight of 2
dogs/sex at 24 months and the average consumption [mean of 22 monthly
measurements], these doses correspond co approximately 311, 1,013 and 4,070
tug/kg/day, respectively, for males and 344, 1,034 and 4,576 mg/kg/day,
respectively, for females). Two addielonal control groups were run concurrently
for each sex and were either untreated or received 10Z cotton linters (cellulose
linters; approximately 2,888 and 3,874 mg/kg/day, males and females, respectively,
when calculated as above) in their diets. No toxic signs related to NC intake
were seen in any of the dogs ac any time.
Following 12 months of treatment without a recovery period, one male and one
female per treatment group were sacrificed. Hlstopathological examination
revealed a variety of "minimal to mild lesions" in all. the dogs examined and
moderate thyroid hyperplasia in a control and a high dose female and moderate
thymus involution in a cotton control male. These lesions included microcalculi
in the medula of the kidney, parafollicular cell hyperplasia of the thyroid,
hemosiderosis of the spleen and axillary lymph nodes and involution of the thymus.
The lesions were considered by the authors to be typical of those naturally
occurring in dogs of the age tested and were, therefore, not considered
treatment-related. Since there were no effects related to NC treatment,
histologic data from the low and middle dose dogs were not evaluated, and the
necropsy on the dogs in the recovery group (one/sex/treatment group) was omitted.
No dose-related variations were reported for routine hematologic parameters nor in
fasting blood glucose. serua transaminases (SCOT, SGPT), cholesterol, alkaline
phosphatase sad blood urea nitrogen (BUM). The body weights, absolute and
relative orgaa weights were comparable to unexposed controls or cotton controls
(10Z linters). However* there was a dose-related Increase In feed consumption.
Similarly, dog* (2/sex/treatment group) adalnlstered NC (up to 10Z) for 24
months (without s recovery period) showed what the authors considered naturally
occurring histopathologic lesions. These lesions Included pituitary cysts, mild
to severe chronic lymphocytic thyroiditis, pleural fibrosis, muscular hypertrophy
of bronchioles, mild to moderate granulomatous pneumonia. renal calcinosis,
extramedullary hematopolesls of the spleen and hemosiderosis of the lymph nodes.
The absolute and relative orgaa weights had sons variations between individual
dogs, but no apparent differences between dogs in different treatment groups.
V-4

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3ecause there were no observable effeccs froa feedirg N'C for 24 re-.:-.s, :-e
necropsy on the dogs in the recovery group (2/sex/creacT.enc group) was cz.zzzd.
Male and female racs (32/sex/treatment group) were similarly exposed to NC ac
I", 3Z, or 10Z In the feed on a dry weight basis for up to 24 months (Ellis et
al.t 1980). Controls, similar to those used in dog studies, were run
concurrently. A separate group of raps, 8/sex/group, dosed as above, was started
6 months later and used for the one year necropsy and a 4-week recovery study.
(Ref. Table Al-l: Using the authors' data for average terminal body weight at 24
months and the average consumption over 24 months [mean of 23 monthly
measurements], these doses correspond to approximately 350 and 1,280 mg/kg/day for
1Z and 3Z, respectively, in males and to 373 and 1,422 mg/kg/day, respectively, in
females.) Apparent feed intake for rats given 10Z NC or 10Z cotton linters was
approximately twice by weight that of untreated controls. However, the authors
Indicated that these two groups had visible scattering of feed and fiber around
the cages. This scattering would account for part of the loss of weight in the
feeders (the parameter measured) and, therefore, the apparent increased intake
(actual intake of NC or cotton linters at 10Z in the diet) cannot be calculated
with any degree of accuracy.
Following 12 months of treatment, all exposed rats shoved a dose-related
increase in food consumption. High dose and cotton control (10Z linters) male
rats failed to gain weight or lose weight in the first week and, thereafter,
gained weight more slowly than the controls. The authors felt that this effect
was consistent with the face that NC serves as non-nutritive dietary bulk. Other
than normal species variations, no significant effects ware apparent for routine
hematologic parameters nor for fasting blood glucose, SCOT, SGPT, cholesterol,
alkaline phosphaeasa and BUN. Absolute and ralaclva organ wcighes were generally
comparable to controls with a few statistically significant differences (decrease
in testes ac 3Z and spleen ae 3Z and 10Z, linters and controls in males). The
authors reported thesa affects as normal variations. Upon histologic examination,
mild to moderate chronic murine pneumonia was found in all unereaeed control rats,
cotton control rats, and tha NC-treaced rata at ehe high dose levels. A variety
of other lesions was found in ehase rats including bile duct hyperplasia and mild
to severe nephritis In aalea only and cystic degeneration of the adrenal gland,
and excessive hesoslderin in tha spleen of the females only. ' Tracheitis was
wide-spread and slid to severe in nature. One 10Z cotton linters female displayed
a fibroadenoma of the aaomary gland. Since the authors found no apparent
relationahlp between NC feeding and the incidence or severity of any of these
lesions* histologic data for low and middle dose rats were not evaluated and the
necropsy after one month of recovery was omitted.
It was further reported that in rats treated for 24 months, there were no
statistically significant differencea in absolute and relative organ weights
between treatment groups or when compared to controls. Upon histopathologic
exaalnation, a large nuaber of lesions were found in various tissues of rats
fed NC for 24 months without a recovery period but none were reported to be
V-5

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significantly different froa controls. Some lesions, such as chronic r_r.-s
pneumonia, were found in most rats. Other lesions were found less _f requer.:. •
the variety and incidence of these lesions attributed co being t>pical of
geriatric rata. The lesions included bile duct hyperplasia, foci or areas o:
hepatocellular alteration, testicular degeneration and/or atrophy, senile
nephropathy, nephritis or focal tubular nephrosis and mammary gland fibroadenoma,
adenoma or adenocarcinoma. Hemosiderosis and/or extramedullary hematopoiesis of
the spleen occurred almost twice as frequently in females than in males while
myocardial degeneration and/or fibrosis were twice as frequent in males. Mo
lesions were reported to be related, in Incidence or severity, to the NC dose
administered. Similarly, lesions from rats fed NC for 24 months and allowed to
recover for 1 month vera not considered to be NC-related. Also, in rats dying
before the scheduled termination of the experiment (averaging between 40Z to 457.
in males and females, respectively), there were no NC-related lesions reported.
The cause of these early deaths was not addressed by the authors.
Ellis et al. (1980) also studied the effects of NC administration in male and
female mice (CD-I strain; 58/sex/group) at 1Z, 3Z or 10Z in the feed on a dry
weight basis for up to 24 months employing procedures similar to those used for
dogs and rats. Controls received either 10Z cotton llnters in the diet or regular
chow. (Ref. Table Al-l: Using the authors' data for average terminal body weight
at 24 months and the average consumption over 24 months [mean of 18 monthly
measurements; measurements for months 4 to 7 vara omitted due to excessive
scattering of feed ae all levels], these doses correspond to approximately 1,814
and 4,866 mg/kg/day at 1Z and 3Z, respectively, for males and 1,767 and 6,056
mg/kg/day, respectively, for females). As In the rat study previously described,
the actual Intake of 10Z NC or 10Z cotton llnters in the diet could not be
calculated with any degrea of accuracy due to what was described as an excessive
scattering of tha fibers.
As reported In tha previous study by Ellis et al. (1976), some early deaths
(in tha first 3 weeks of the study) dua to Intestinal impaction by the fibrous
material also occurred In miea fad tha high dosa (10Z) of NC and cotton control
(10Z llnters) for up to 24 months. Gross necropsy found emboli of white fibrous
material blocking tha inesstinas at various sites from tha jojunum downward.
Additional mica, fad ths respective diets from tha start of tha study, were
substltutad for th« dead mica.
Tha hltffc flbsr contane of tha 10Z diets (cotton lintars and NC) was also
reported am chs probable causa of an apparene irritation effece in these animals
that firsr occurred la week 18 and decreased by tha end of month 10. This effect
was charactarlxed by hyperemia of tha ears and eyelids, and subsequent edema of
tha genitalis* fast and call accompanied by contlnuad scratching. This effect was
not correlaced with any other toxic sign end was almosc non-existent in the lower
treatmane levela and untreated controle. The effect was, therefore, attributed to
tha physical, not chemical, nacura of the dlee.
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In aice fed N'C for 12 -norths, the only scaciscical differences a-o-g ;<-e
treatment groups (4/sex/level) were significant increases in weight of cne spLeen
and ovaries, both absolute and relative to body and brain weight, in che cocton
control females. Since similar increases were not seen at other necropsies nor
were pathological lesions found, these increases were presumed by the authors to
be normal variations, unrelated to treatment. Animals fed the 10Z diets (NC or
linters) lost weight during the first week of the study but quickly began to gain,
leveling off by A months, with average weights of all dosage groups converging
during the second year. Upon histologic examination, male and female mice fed N'C
at all levels for 12 months shoved a variety of lesions, usually degenerative or
inflammatory. As is commonly seen in older mice, widespread amyloidosis of most
tissues was observed, with deposits most heavily concentrated in the intestine and
kidney. However, the authors indicated there was no consiseene relationship
between any of the lesions and the NC treatment. Also, no treatment-related
variations in the results of routine hematologic parameters, SGPT or BUN, were
reported.
In mice fed NC for 24 months with or without a recovery period, there were no
treatment-related differences in organ weights, routine hematof&glc parameters,
SCOT, SGPT nor BUM. Tissue lesions were found in mica fed the untreated control
diet or high (10Z) NC dose and sacrificed after 24 months feeding, as veil as in
those animals dying before that time. A variety of lesions, usually degenerative,
were seen. Amyloid deposles, often heavy, were widespread. These were noe
reported to be related to NC exposure. No histologic data were provided for
control mice fed 10Z cotton linters. Due to a lack of ereatment related effects,
no histologic data were provided for mice fed NC at 1Z and 3Z; nor were these data
provided for treated mice allowed to recover for one month following NC treatment.
At about month 9, many deaths (approximately 2SZ of the original number) with
no apparent cause occurred in the high dose mice with a smaller number occurring
in the cotton controls. A physical fiber effect was considered; however, since
the death rata was three times higher In the NC-treatad mice, the authors felt
that a compound effect of unknovn mechanism could not be dismissed. Rapid
autolysis characteristic of aice precluded histopathological pxamlnatlon.
C. Genetic Toxicology
Studies have been conducted to determine the mutagenic potential of
nitrocellulose employing bacterial assays as well as cytogenetic tests. Mutagenic
activity was not detected using these bloassay systems under conditions of the
tests.
Ellis et al. (1978) conducted an Ames test exposing typhlmurlum tester
strains TA1535, TA1537, TA1538, TA98 and TA100 to nitrocellulose at 100, 1,000 or
5,000 mlcrograas/plata for 48 hours. No statistically significant Increase in the
number of revertant colonies was observed when compared to levels of spontaneous
revsrtants either with or without S9 metabolic activation (added rodent liver
homogenate).
V- 7

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Ellis et al. ( 1976) studied the cytogenetic effects of \C on sonacii. cel.
chromosomes employing peripheral blood lymphocyte and kidney cultures from racs
fed NC at 10Z (mg/kg intake cannot be calculated due to excessive scatterirg of
fibers) for 13 weeks. Treated rats did not show any changes in chromosome
frequency distribution, number of tetraploids, or frequency of chromatid breaks,
gaps or translocation in the test cultures when compared to cotton control (10"
linters) cultures. No data on untreated controls was available for comparison.
Ellis et al. (1980) reported that kidney and bone marrow cells from rats exposed
to NC or cotton linters in the diet at 10Z (exact dose not calculated due to
excessive scattering) for 2 years showed no numerical or morphological changes in
chromosomes.
D. Carcinogenicity
Long-tern studies conducted in dogs, rats and mice indicated that the
administration of NC in Che diet for up to 24 months did not significantly affect
the incidence of tumors in various organs/systems in dogs, rats and mice when
compared to control animals.
Ellis et al. (1980) conducted 12 or 24 month feeding studies to determine the
incidence of tumor formation following exposure to NC at up to 10Z in the diets of
dogs, rats and mice. The studies described belov are the same as those described
under "Lifetime Exposure" and are discussed more fully therein.
Beagle dogs (6/sex/dose) were administered 1Z, 3Z or 10Z NC (Ref. Table Al— L:
These doses correspond to approximately 311, 1,013 and 4,070 mg/kg/day,
respectively, for males and 344, 1,034 and 4,576 mg/kg/day, respectively, for
females, as described under Lifetime Exposure) for 12 to 24 months. Two
additional groups were run concurrently for each sex and either remained untreated
or received 10Z cotton linters in their diets. Following treatment, the animals
were sacrificed and tissues were examined for occurrence of tumors in various
organ systems. The authors reported no biologic variations or tumors in the test
dogs.
Rats, 32/sex/dose* were fed NC for up to 24 months at II, 3Z or 10Z in the
diet (Ellis et al., 1980). (Ref. Table Al-1: These levels correspond to
approximately 330 and 1,280 mg/kg/day for 1Z and 3Z, respectively, in males and
373 and 1,422 mg/kg/day, respectively, In females. Intakes for the 10Z level
could nor be calculated.) Two additional groups were run concurrently for each
sex and either vara untreated or received 10Z cotton linters. Following
treatment, tha animal* ware sacrificed and evaluated histologically for tumor
formation in various organ systems.
The authors rsported the occurrence of pituitary chromophobe cell adenomas
and various mammary tumors. Including fibroadenomas, adenomas, and
adenocarcinomas/carcinomas at all dose levels. Thssa tumors also occurred in
untreated and cotton controls. There waa a scattered incidence of many other
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naturally occurring tumor types In rats in all treatment and control grcu;s.
lesions found ia various organ systems in SC-treated rats were attributed b .• ;-e
authors to nitrocellulose.
Ellis et al. (1980) also fed three groups of CD-I mice (58/sex/group) sc at
IZ, 3Z or 10Z as dry weight in the feed for up to 24 months. (Ref. Table a 1-1:
These levels correspond to approximately 1,814 and 4,866 mg/kg/day at IZ and 37.,
respectively, for males and 1,767 and 6,056 mg/kg/day, respectively, for females.
Intake for the 10Z level could not be calculated.) Two additional groups were run
concurrently for each sex and either were untreated or received 10Z cotton
linters. Ac the end of the treatment period, the control and treated mice were
sacrificed and tissues ware examined for occurrence of tumors in various organ
systems.
The authors reported the occurrence of bronchloalveolar carcinomas in control
animals, but none vera observed In mice administered 10Z NC for 24 months. The
absence of such tumors was statistically significant in males but not in females
(P ¦ 0.004 for males, P ¦ 0.29 for females, P • 0.002 for the combined sexes, by
exact analysis of the contingency table). However, the authors concluded that the
difference probably represents natural variation and is toxlcologlcally
meaningless. The slides from other dosage groups were not examined.
E. Reproductive Effects
Three-generation studies in rats indicated that NC administered alone in the
diet at dose levels up to 10Z did not adversely affect reproduction. However, the
increased inert bulk of 10Z fibers (NC or linters) may reduce the lactation index
and pup velghe.
Ellis et al. (1980) administered nitrocellulose in the diet of rats at 1Z, 3Z
or 10Z in a three-generation study. (Ref. Table Al-1: These doses correspond to
approximately 350 and 1,280 mg/kg/day for 1Z and 3Z, respectively, in males and
373 and 1,422 mg/kg/day, respectively, for females. The actual intake for the 10Z
dose level cannot be calculated.) The initial groups of rats used as the parental
(F ) generations were started at the sama time as the chronic study. Rats of each
subsequent group* parents and offspring, received the sama control or
NC-containlag dlata as their original PQ generation. First mating occurred when
rats wera 8- aontha of aga. Following treatment, all offspring were examined at
birth for gross physical abnormalities and the numbers of live and dead pups of
each litta* vara racordad. Survival and body weighta wara recorded at 0, 4 and 21
days. Reproductiva performance for each parental and offspring generation was
determlnad.
The authora reported that the mean body weights at the time of first matlngs
for males of all parental generations given diets containing 10Z NC were
significantly reduced whan compared to males given untreated control diet. This
reduction in body weight waa also significant in males and females fed cotton
control (10Z linters) diets.
V-9

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There were no indications chat the treacrents adversely a::e;ced :~a
fertility of the males or females in the mating or pregnancy ratios. Fcr :-e "
generation, most of these parameters actually suggested that either 102 NC or M1"
cotton linters in the diet increased the fertility of rats given these treatrencs.
However, this effect appears to be caused by the decreased fertility of the
control Fq females. The fereilicy of the F^ females was similar to that of
controls For groups given 1Z and 3Z NC (approximately 373 and 1,422 mg/kg/day).
This decreased fertility in the control and low dose groups was attributed by
the authors to both the age at time of first mating (8 months for all F parental
generations) and body weights greater than that expected to give optimal
reproductive performance.
No treatment-related effects were apparent at any dose or in any generation
on litter size, liveborn Index, birch weight, viability index, or the ratio
of males to total offspring. Significant reductions In the lactation index and
the weight of pups at weaning occurred with 10Z NC and 10Z cotton llnter controls.
Reductions in these parameters were observed chiefly with the F through F
litters for both groups but were not observed with the subsequent litters. 2^he
authors attributed these reductions during lactation to relatively large amounts
of inert bulk in a diet of 10Z fibers (NC or llnters).
F.	Teratogenicity
Ellis et al, (1980) did not report conducting teratology studies.
G.	Immunologic Effects
Ellis et al. (1976) studied the potential adverse effects of NC exposure on
the immunologic response based on serum concentration of immunoglobulin (IgE)
measured by the Immunodiffusion technique of Manclnl et al. (1964). The
administration of 10Z NC la the feed to dogs or rats (this dietary level can be
calculated, as described under Thirteen-Week Studies( to correspond to an average
of approximately 7,690 mg/kg/day for dogs but could not be calculated for rats due
to excessive scattering of fibers) for up to 13 weeks did not alter their serum
concentrations of IgE*
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VI. health advisory development
Nitrocellulose, at doses averaging 4,300 ag/kg/day in dogs and in excess c:
approximately 1,400 mg/kg/day in racs and 6,000 ng/kg/day in mice, was not coxic
when fed for up to 24 months. The actual intakes in mg/kg/day in rats and mice
fed 10Z NC could not be determined with any degree of accuracy due to visible
scattering of the fibers around the cage. Assuming, however, that these animals
consumed only tvice the amount of NC as those receiving the 3Z diet, a reasonably
conservative estimate, Intake would be, at a minimum, approximately 2,800
mg/kg/day in rats and 12,000 mg/kg/day in mice. The only treatment-relaced
effects in these high-dose,, long-term feeding studies were early weight loss in
rats and mice and subsequent lower average body weights in the mature rat,
hyperemia with edema of the ears, eyelids, genitalia, feet and tall apparently due
to physical-mechanical effects of the fibers and death due to intestinal impaction
in mice fed the 10Z diets (NC and cotton linters).
The weight effect was attributed to the non-nutritive bulk of the fibrous
diet and was confirmed by similar findings In control animals fed 10Z cotton
linters (cellulose linters), the material that Is nitrated to form NC. This lower
body weight, however, is not necessarily an adverse effect as the rodents fed the
10Z diets were characterized by less fat, not less lean body mass, when compared
to untreated anlaals of the same species. This condition can, however, be
detrimental If there la a very high body demand for nutrients, as during pregnancy
and lactation. This condition occurred, to soma degree, in the early litters of
the three-generation reproduction study in rats, as evidenced by a decreased
lactation index and weight at weaning, but not in later generations, possibly
indicating adaptation.
The hyperemia with edema that occurred in mice was not life-threatening,
resolved spontaneously and was of unknown cause. As this effect occurred in mice
fed either 10Z NC or cotton linters and was accompanied by continuous scratching,
a direct physical effect dua to irritation from contact with the fibers seems
likely.
Tha deaths dua to Intestinal impaction vara attributed 'to the relatively
large slzs of eh* fibers In relation to tha size of tha luaen of the Intestines,
allowing eh* asssas fomed by tha fibers to completely obstruct the gut. This
effect occusrad la mica fad both the 10Z NC and cotton linters. Unresolved is a
spate of death* of unknown causa that occurred at 9 months in both tha 10Z NC and
cotton llat*rs groups but was at a higher level in the NC group. This may be due,
In pare* e© tha chemical nature of NC.
In 13-week feeding seudles ae tha eeae levels. Intakes averaged 7,700
mg/kg/day In dogs and were In exceaa of approximately 2,700 mg/kg/day in rats and
7,000 mg/kg/day In mica. Calculation of tha 10Z levels to approximates, as in the
long-term study, would yield values, et e minimis, of 5,400 ng/kg/day la rats and
14,000 mg/kg/day In alee. There waa no evidence of toxicity. The Increased food
VI-1

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consumption/decreased weight gain pattern in rats and 21.ce was similar :o z-.az -
the Z^-month study, occurred at the 10T level in amr.als fed either S'C or c::ccr
linters and was attributed to the non-nutritive bulk of the fibers.
Death due to intestinal impaction also occurred in the high-dosed mice in
this study and was.attributed to the physical nature of the fibers. Since the
treated animals, like humans, cannot digest cellulose, passage of these fibers
through the digestive tract would be expected.
This non-digestion of the fibrous diet was confirmed in absorption studies.
After repeated oral doses (once daily for four days) of radiolabeled
nitrocellulose to rats, no detectable radioactivity was found in'any tissue or
body fluid but was recovered only from various components of the GI tract,
contents and feces.
Nitrocellulose was noe mutagenic in either bacterial bloassays nor did it
produce chromosome abnormalities in mammalian cells after in vivo exposure. It
did noc significantly affect tumor incidence in dogs, rats and mice and produced
only some adverse effects la rats with high nutrient requirements during pregnancy
and lactation.
In view of the non-eoxic nature of NC ae all doses seudled and its failure to
be digested and absorbed In ch« species tested, health advisory (HA) values for
1-Day, 10-Days and Longer-term appear to b« unnecessary. Ie seems probable that,
due to the fibrous nature of Che substance and Its insolubility in water, clarity
or turbidity of tha water would be the only guideline necessary.
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VII. ANALYSIS
Rosenblatt et al. (1973) in summarizing literature on the analysis of \C
indicated that all analytical procedures probably would begin with collection of
NC fibers from the water on filters. Weighing the filter is cited as a method co
roughly estimate NC, but its accuracy would be limited by the presence of ir.erc
solids.
Other methods cited include:
o Ferrous-titanous titration
o Ferrous sulfate titration
o Liberation of NO^ gas
o Analysis of NH^ after reduction by Devarda's alloy
o Transnitratlon of salicylate or citrate followed by
ferrous-titanous titration
o Chromous chloride-ferric ammonium sulfate micro-determination
o Zinc dust reduction of the nitrate ester
o Hydrolytlc liberation of nitrite ion in acetone
Of the detection methods listed above, the last one is considered to be the
most effective for detecting lov levels of NC In the environment (Sullivan et al.,
1978; Rosenblatt et al., 1973). It Is a colorlmetrlc method based on hydrolytic
liberation of nitrite by OH from acetone solutions of nitrate esters. The
resulting NO^ is then dlazotlzed with either alpha-(naphthyl)-ethylenedlaolne
hydrochloride or alpha-naphthylamine and the absorbance of the solution determined
at 520 to 530 nm. The reaction Is not specific for NC; however, the insolubility
of this compound In water allows quantitative separation from NO^, NO and
other soluble nitrate esters or nitrocompounds In mixed wastewater by filtration
or dialysis. Berkley and Rosenblatt (1978) adapted this procedure to the
Technlcon Autoanalyzer. The procedure involves aspiration of a stirred NC
suspension, dialysis against 9Z saline, and hydrolysis with 5N NaOH at 70s C for
10 minutes to release nitrite ion. Sulfanllle acid Is dlazotlzed by the nitrite
ion at lov pH. The resulting dlaxonlua salt Is coupled with the
N-(l-naphthyl)ethyleaediaaina, and the color produced Is measured at 520 nm. The
limit of detactability la 0.4 ag/L of nitrocellulose.
Sullivttet al.(1978) outlined two methods for determining NC in sediment.
The first aathod Involves solvent extraction of the dried sediment In acetone or
ethylacetaeas. The procedure la sensitive to as little as 0.5 mg/kg of nitrate
ester; hovavar, It is not specific for NC because le also extracts other organic
nitrate esters and nitrocompounds. The second method Involves an initial acetone
extraction to reaove the nitrate ester. Nitrate Is then determined
colorimetrlcally since le will oxidize ferrous Iron to ferric Iron after treating
the extract with acetic acid, ferrous sulfate In sulfuric acid, and sodiua
sulfite. Absorbance of the resulting yellow color is determined at 500 nm. This
procedure, however, lacka sensitivity since the minimum detectable concentration
in a 10 g sediment sample is 140 ag/kg.
VII-1

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VIII. TREATMENT
Wastewater from NC production facilities is neutralized, then settled,
centrifuged, and/or screened to recover NC fibers (Sullivan et al., 1978).
Cencrifugation leads to more efficient and constant recovery due to the high
specific gravity of NC (Rosenblatt et al., 1973).
The US Army Natick Laboratories (Massachusetts) has developed a chemical and
microbiological process for the degradation of NC in wastewater (Rosenblatt et
al.t 1973). The process Involves membrane ultrafiltration of the wastewater to
concentrate the suspended NC. A 200-fold concentration, to a 3Z to 5Z NC
suspension, was desired; however, only a 10-fold concentration, to 0.2Z NC, has
been obtained by this process. The suspension is then treated with 3Z NaOH ac 90°
to 95* C for 20 minutes to yield a soluble material containing little nitrate
ester. After acid neutralization, nitrate escer content of the filtrate is
determined by IR analysis. An extract of tha filter is similarly analyzed to
detect any undissolved nltrata ester, presumably NC. The neutralized solution is
mixed with domestic waste and fermented anaeroblcally to denitrify thus producing
gaseous nitrogen as a produce. Methanol could be added as a nutrient in this
step. The next step is an aerobic activated sludge treatment and finally, a
second denltrification, again using methanol nutrient. The final product was
reported to be nitrocellulose free.
VIII-1

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IX. CONCLUSIONS A.VD RECOMMENDATIONS
Based on available toxicity data and the chemical and physical properties of
the compound* NC is apparently non-toxic to dogs, rats and mice and is noc
digested nor absorbed in these species. These data, along with the relative
insolubility of NC in vater, suggest that Health Advisory values for NC in
drinking vater are unnecessary. The physical characteristics of the drinking
vater as they relate to turbidity, clarity, caste and similar indicators of
palatability appear to be the only guidelines necessary.
A companion report, "Data Deficiences/Problem Areas and Recommendations for
Additional Data Base Development For Nitrocellulose" (Appendix 2) summarizes the
scope and adequacy of existing data reviewed for this HA and delineates those
areas where additional data, if any, are deemed necessary.
IX-1

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X. REFERENCES
Barkley, J.J. and Rosenblatt, D.H., 1978. Automated nitrocellulose analysis.
U.S. Army Medical Bioengineering Research and Development Laboratory Technical
Report 7807, AD A067081, Fort Detrick, MD.
Department of the Army Technical Manual TM9-1300-214, 7-7. 1967. Departments of
the Army and the Air Force. Military Explosives. Nitrocellulose.
Ellis, H.V., III et al., 1976. Mammalian toxicity of munitions compounds Phase
II: Effects of multiple doses Part IV: Nitrocellulose. Report No. 5. 'Midwest
Research Institute. Kansas City, MO, Contract No. DAMD-17-74-C-4073, AD A062016.
Ellis, H.V., III et al., 1978. Mammalian toxicity of munitions compounds Phase I:
Acute oral toxicity, primary skin and eye irritation, dermal sensitization,
disposition and metabolism, and Ames tests of additional compounds. Report No. 6,
Midwest Research Institute. Kansas City, MO, Contract No. DAMD-17-74-C-4073, AD
A069333.
Ellis, H.V., III et al., 1980. Mammalian toxicity of munitions compounds Phase
III: Effects of life-time exposure Part III: Nitrocellulose. Report No. 9.
Midwest Research Institute, Kansas City, MO, Contract No. DAMD-17-74-C-4073.
Finney, O.J., 1971. Probit Analysis, Cambridge University Press.
Hawley, G.G., 1981. The condensed chemical dictionary. Tenth edition. Van
Nostrand-Reinhold.
Helton, D.O., 1976. Chemical and physical characterization of nitrocellulose
fines. Special Report. Midwest Research Institute, Kansas City, MO, Contract No.
DAMD-17-74-C-4073.
Huff, B.L. et al., 1975* Aquatic field surveys at Radford, Holston, Volunteer,
and Milan Army anunition plants. Volume 1-Radford. Final Report. Wapora, Inc.,
Washington, D.C., Contract No. DAMD-17-74-C-4138.
Lee, C.C. et al., 1975. Maanallan toxicity of munition compounds: Phase I. Acute
oral toxicity* primary skin and eye irritation, dermal sensitization, and
disposition and metabolism. Report No. 1. Midwest Research Institute, Kansas
City, MO» Contract No. DAMD-17-74-C-4073, AD B011150L.
Mancini, G. et al., 1964. Immunochemical quantitation of antigens by single
radial immunodiffusion. Isaunocheaistry, 2:235.
Rosenblatt, D.H. et al., 1973. Munitions production products of potential concern
as waterborne pollutants-Phase I. U.S. Army Medical Environmental Engineering
Research Unit Report No. 73-07, Edgewood Arsenal, MD.
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RTICS, 1986. Registry of Toxic Effects of Chemical Substances, Natioral Ir.sc:t-:e
for Occupational Safety and Health (NIOSH). National Library of Medicine Online
File.
Selig, V., 1961. Microdetermination of aromatic nitro compounds, nitrocellulose,
and cyclic nitramines. AEC Report l'CRL-6639:20-28.
Sullivan, J.H. et al., 1978. A summary and evaluation of aquatic environmental
data in relation to establishing water quality criteria for munitions unique
compounds. Part I. Nitrocellulose. Final Report. U.S. Army Medical Research
and Development Command Contract Number DAMD-17-77-C-7027, Water and Air Research,
Inc., Gainsvlllc, FL.
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APPENDIX 1
CALCULATION METHODS
AI — 1

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IA3LE A1- 1
Calculation of Intake of Test Material (mg/kg/day)
Intake • (a)(b)(1000)
(100)(c)
where:	a •	X NC In dlec
b -	intake of feed In grama/day*
1000 ¦	conversion factor, grams to milligrams
100 ¦	conversion factor, Z to grams/100 grams
c -	animal weight in kilograms*
*Intake/animal weight data used may vary for each calculation depending upon
information available in report. Text will describe exact conditions for
determining these calculations for each experiment.
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APPENDIX 2
DATA DEFICIENCIES/PROBLEM AREAS AND RECOMMENDATIONS FOR
ADDITIONAL DATA BASE DEVELOPMENT FOR NITROCELLULOSE
A2-1

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TA3LE OF CONTENTS
P-.CZ
INTRODUCTION 	 A2-3
OBJECTIVES	A 2-3
BACKGROUND	A2-3
DISCUSSION	A2-4
CONCLUSIONS/RECOMMENDATIONS 		A2-5
REFERENCES	A2-6
A2-2

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INTRODUCTION
The Office of Drinking Water (ODW), Environmental Protection Agency (EPA), .a
conjunction with the Department of the Army, has reviewed the available data on
nitrocellulose (NC) for the purpose of developing a Health Advisory (HA) useful -.a
dealing with contamination of drinking water, to include "state-of-the-art"
information on health effects, analytical methodology and treatment technology.
This information is contained in detail in the report entitled "Health Advisory on
Nitrocellulose."
OBJECTIVES
The objective of this document is to provide an evaluation of data
deficiencies and/or problem areas encountered in the review process for NC and to
make recommendations, as appropriate, for additional data base development. This
document is presented as an independent analysis of the currene status of NC
toxicology, as related to its possible presence in drinking water, and includes a
summary of the background information used in the development of the HA. For
greater detail on the toxicology of NC, the original "Health Advisory on
Nitrocellulose'* should be consulted.
BACKGROUND
Nitrocellulose is a non-volatile, fibrous, cotton-like, white solid used as a
principal ingredient of propellents, smokeless powder, rocket fuel, mortar
increments and some explosives (Sullivan et al., 1978). It is produced for
military use at selected Army ammunition plants (AAP), and NC fines are found in
production wastewater (Huff et al., 1975). It Is resistant to biological
degradation and is* therefore, persistent In the environment and deemed a
potential, if not known, contaminant in drinking water.
The metabolism of NC as studied In rats (Ellis et al., 1976) Indicated that
the compound remained in the gastrointestinal (GI) tract and was excreted
unchanged.
j
Acute toxicity studies in rats and mice (Lee et al.* 1975) indicated that NC
was nontoxic* with Upvalues greater than 5*000 mg/kg reported In both species.
Thlrtcm-iraek (Ellis et al.. 1976) and 24-month (Ellis et al., 1980) feeding
studies is threo specie* (dogs* rats and mlcs) gave no evidence of adverse or
toxic effect* related to the chemical characteristics of NC; nor was NC found to
be carcinogenic. The fibrous physical nature of NC. however* was the cause of
early deaths duo to intestinal impaction In mice fed 10X NC or cotton linters in
both tho 13-week and 24-aonth studies and was associatsd with an Increesed food
lntske-decreaaed weight gain pattern in high dosed and cotton control rats and
mica. The fibers were also reported to be the ciuse of a physical Irritation of
the extremities of mica fed 102 NC or cotton llnter* aa evidenced by hyperemia and
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edema. A spate of deaths of unknown cause occurred in aice after nine -c-:-s o:
feeding NC or cotton linters at 10Z in che diet. The number of deaths was his-er
in che NC treated mice and may, therefore, be related to the chemical nature of
the compound via a mechanism, as yet, undetermined (Ellis ec al., 1980).
Nitrocellulose was reported to be non-mutagenlc in various indicator systems
(Ellis et al., 1976, 1978, 1980). Three-generation reproduction studies in rats
indicated thac NC did not adversely effect reproduction but that che non-nutritive
bulk of the 10Z diet may cause an adverse effect during periods of high
nucritlonal demand, such as pregnancy or lactation, as evidenced by a decrease in
lactation index and weight at weaning (Ellis et al., 1980).
No teratogenic studies were reported. Exposure to NC did not affect the
immunological response of dogs or rats (Ellis «t al., 1976).
Methods of analysis (Barkley and Rosenblatt, 1978) and treatment (Rosenblatt
et al., 1973) adequate for detection and removal of NC at levels which mighc be
deemed hazardous to health have been presented in detail in "Health Advisory on
Nitrocellulose."
The lack of toxicological indicators, along with the apparent
non-absorptlon/non-dlgestion of tha NC fibers, led to the conclusion that HA
values for NC in drinking water wera unnecessary.
DISCUSSION
Available data on the pharmacokinetics, health effects, analysis and NC
wastewater treatment have been reviewed.
While the available data on tha metabolism of NC are limited in scope, the
chemical and physical nature of NC generally supports tha finding of tha one
available study that NC passes through tha G1 tract apparently unchanged and
unabsorbed. Additional studies would, therefore, ba unnecessary.
Tha available studies on tha toxicity of NC include LD 's in rats and mice
and short-tars (13 week) and longer-term (24 month) atudles in dogs, rats and mice
that included asaesaaents for possible carcinogenicity. Three-generation
reproductionatudle* in rats, mutagenicity aasaya in bacterial and cytogenetic
systems an£lMBmologlcal studies in dogs and rats have also bean reported. All
studies appear*adequate for use in HA development.
Further lnveetigatloa of the causa of the spaea of deaths in mice occurring
at nine months in the longer-ten feeding study sees warranted but, in vi*w of che
otherwise low toxicity of >C along with the other available data, would not be
necessary for HA developaea*. Teratogenic studies were not reported; however, tha
apparent inability of MC to be absorbed would preclude its acting as a potential
teratogen. Therefore, additional studies seem unnecessary at this time.
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Primary skin and eye irritation tests in rabbits were negative (Lee a: * •_
1975). However, mice fed 10X N'C or cotton linters for up to 24 concns de-.el—ad
edema and hyperemia of the extremities, apparently due to the fibrous nature of
NC. More sensitive dermal studies, if available, would be appropriate but vculd
~ot be deemed necessary as regards contamination of the drinking water by NC.
Several methods of analysis of NC in wastewater have been reported including
a method adapted co the Technicon Autoanalyzer. The limit of detectability,
0.4 mg/L, appears adequate for determining drinking water contamination relative
to palatability concerns.
Methods for the treatmene of wastewater by chemical and microbiological
degradation have been developed. The insolubility of NC in water and its ability
to be removed by coagulation and/or filtration make it unlikely that more
extensive treatment measures would be required.
CONCLUSIONS/RECOMMENDATIONS
Based on the above discussion, the following conclusions/recommendations can
be made:
1.	The available studies on NC toxicity are adequate for development of a
HA useful in dealing with contamination of drinking water.
2.	No significant data gaps or problem areas relative to NC in drinking
water exist.
3.	No further studies on NC, as relates to its possible presence in
drinking water, are necessary at this time.
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RETICENCES
Barkley, J.J. and Rosenblatt, D.H., 1978- Automated nitrocellulose analvsis.
I'.S. Army Medical Bioenglneering Research and Development Laboratory Technical
Report 7807, AD AQ67081, Fort Detrick, MD.
Ellis, K. V., III ec al., 1976. Mammalian toxicity of munitions compounds Phase
II: Effects of multiple doses Part IV; Nitrocellulose. Report No. S. Midwest
Research Institute, Kansas City, MO, Contract No. DAMD-17-74-C-4073, AD A062016.
Ellis, K.V., III et al., 1978. Mammalian toxicity of munitions compounds Phase I:
Acute oral toxicity, primary skin and eye irritation, dermal sensitization,
disposition and metabolism, and Aaes tests of additional compounds. Report No. 6,
Midwest Research Institute, Kansas City, MO, Contract No. DAMD-17-74-C-4073, AD
A060333.
Ellis, H.V., III et al., 1980. Mammalian toxicity of munitions compounds Phase
III: Effects of life-time exposure Part III: Nitrocellulose.p Report No. 9.
Midwest Research Institute, Kansas City, MO, Contract No. DAMD-17-74-C-4073.
Huff, E.L. et al., 1975. Aquatic field surveys at Radford, Holston, Volunteer,
and Milan Army ammunition plants, Volume I-Radford. Final Report. Vapora, Inc.,
Washington, D.C., Contract No. DAMD-17-74-C-4138.
Lee, C.C. et al., 1975. Mammalian toxicity of munition compounds: Phase I.
Acute oral toxicity, primary skin and ey« irritation, dermal sensitization, and
disposition and metabolisa. Report No. 1. Midwest Research Institute, Kansas
City, MO, Contract No. DAMD-17-74-C-4073, AD B011150L.
Rosenblatt, D.K. et al., 1973. Munition* production products of potential concern
as waterborae pollutants-Phass I. U.S. Army Madlcal Environmental Engineering
Research Unit Report No. 73-07, Edgewood Arsenal. MD.
Sullivan, J.H. et al., 1978. A suanary and evaluation of aquatic environmental
data in relation to establishing water quality criteria for munitions unique
compounds* Part I. Nitrocellulose. Final Report. U.S. Army Medical Research
and nautili—ill fiwuil Contract Number DAMD-17-77-C-7027, Water and Air Research,
Inc., Gain«viXl«, FL.
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