DTIC ADA545795: Review of Ammonium Dinitramide Toxicity Studies

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by  Defense Technical Information Center

AFRL-RH-WP-TR-2011-0059

Review of Ammonium Dinitramide Toxicity Studies

David R. Mattie
Biosciences and Performance Division
Applied Biotechnology Branch

Teresa R. Sterner
Henry M. Jackson Foundation for the
Advancement of Military Medicine
Wright-Patterson AFB, OH

January 2011

Interim Report for October 2009 to November 2010

Air Force Research Laboratory
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711th Human Performance Wing
release; distribution unlimited.
Human Effectiveness Directorate
Biosciences and Performance Division
Applied Biotechnology Branch
WPAFB, OH 45433-5707

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01-01-2011 Interim 1 Oct 2009 – 30 Nov 2010
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Review of Ammonium Dinitramide Toxicity Studies
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Mattie, David R.*, Sterner, Teresa R.** OAFW
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HJF**
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9. SPONSORING/MONITORING AGENCY NAME(S) AND ADDRESS(ES) 10. SPONSOR/MONITOR'S ACRONYM(S)
Air Force Materiel Command* 711 HPW/RHPB
Air Force Research Laboratory
Human Effectiveness Directorate 11. SPONSORING/MONITORING
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Applied Biotechnology Branch
AFRL-RH-WP-TR-2011-0059
Wright Patterson AFB OH 45433-5707
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Distribution A: Approved for public release; distribution unlimited. Cleared 11 Jul 11, 88ABW-2011-3818.
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14. ABSTRACT
Ammonium dinitramide (ADN) is a class 1.1 explosive oxidizer being considered for use in solid rocket propellant
mixtures. Studies performed evaluating the toxicity of ADN have included an acute and subacute toxicity screen, a 90-
day reproductive screen and three follow-on reproductive studies, electron paramagnetic resonance (EPR) spectroscopy
studies, mutagenicity assays and a study evaluating the effects of ADN on hepatocytes in vitro. The LD in rats is 823
50
mg/kg, indicating ADN is moderately toxic. The results also indicated that ADN is a female reproductive toxicant in rats,
causing implantation failure in early gestation; follow-on studies implied that ADN is embryotoxic. EPR studies indicated
that ADN can decompose to form reactive nitrogen metabolites which can be harmful in biological systems. The
hepatocyte studies suggested that ADN has the potential for directly affecting cellular DNA in vitro; these results are
supported by traditional genotoxicity assays which reported that ADN is mutagenic.
15. SUBJECT TERMS
Ammonium dinitramide, ADN, toxicity, reproductive, embryotoxic, genotoxicity
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U ABSTRACT OF PAGES Timothy W. Bucher

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TABLE OF CONTENTS

List of Tables ................................................................................................................................. iv
Preface..............................................................................................................................................v
Acknowledgements ........................................................................................................................ vi

1.0 Executive Summary ..................................................................................................................1
2.0 Introduction ...............................................................................................................................2
3.0 Toxicity Studies ........................................................................................................................2
3.1 Acute Toxicity ..............................................................................................................2
3.2 Repeated Exposure Study .............................................................................................3
3.3 Reproductive Studies ....................................................................................................4
3.4 Genotoxicity Studies .....................................................................................................7
3.5 Potential Mechanism .....................................................................................................8
4.0 Discussion .................................................................................................................................9
5.0 Conclusions ..............................................................................................................................10
6.0 References ...............................................................................................................................10

List of Abbreviations .....................................................................................................................12

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LIST OF TABLES

Table 1. Results of standard acute toxicity tests with ADN ...........................................................3
Table 2. Results of repeated dose drinking water studies with ADN .............................................3
Table 3. Results of reproductive and developmental studies with ADN ........................................6
Table 4. Results of genotoxicity studies with ADN .......................................................................8

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PREFACE

This review was conducted under the Henry M. Jackson Foundation for the Advancement of
Military Medicine, Inc. (HJF) contract, FA8650-05-2-6518. The program manager for the
contract was Mark Hoffman of the Air Force Research Laboratory, 711 Human Performance
Wing, Human Effectiveness Directorate, Biosciences and Performance Division (711
HPW/RHP). The technical manager for the project was David Mattie, Ph.D. in the Applied
Biotechnology Branch of 711 HPW/RHP.

The animal studies were approved by the Air Force Surgeon General’s Research Human &
Animal Research Panel and the Wright-Patterson Air Force Base Institutional Animal Care and
Use Committee. The studies were conducted in a facility accredited by the Association for the
Assessment and Accreditation of Laboratory Animal Care, International, in accordance with the
Guide for the Care and Use of Laboratory Animals (NRC, 1996).

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ACKNOWLEDGEMENTS

A number of studies for this effort was conducted under the Department of Air Force Contract
No. F33615-90-C-0532. Lt Col Terry A. Childress served as the Contract Technical Monitor for
the U.S. Air Force, Armstrong Laboratory, Toxicology Division. Darol E. Dodd, Ph.D. was the
ManTech Environmental Technology, Inc. program manager for the contract. The efforts were
cosponsored by the U.S. Army Medical Research Detachment, Walter Reed Army Institute of
Research (WRAIR) under the direction of LTC Roland E. Langford, Detachment Commander.
LTC Daniel J. Caldwell served as the technical manager for the Army Medical Research
Detachment. For the studies conducted under this contract, the authors would like to
acknowledge the technical assistance of Marcia L. Freedman, Richard J. Godfrey, Willie J.
Malcomb, Jerry W. Nicholson, Margaret A. Parish, Stephanie A. Salins, SrA Stacie L.
Southwell, SPC Mark Slovik, Major Donald R. Tocco, and Merry J. Walsh.

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1.0 EXECUTIVE SUMMARY

Ammonium dinitramide (ADN) is a class 1.1 explosive oxidizer, which has been repeatedly
evaluated for use in solid rocket propellant mixtures. A number of studies were performed
evaluating the toxicity of ADN, but a review of its toxicity has never been published. Toxicity
studies have included an acute and subacute toxicity screen, a 90-day reproductive screen and
three follow-on reproductive studies, electron paramagnetic resonance (EPR) spectroscopy
studies, mutagenicity assays and a study evaluating the effects of ADN on hepatocytes in vitro.
The LD in rats is 823 mg/kg, indicating that ADN is moderately toxic. No evidence of dermal
50
irritation or general toxicity was found. ADN is a female reproductive toxicant in rats, causing
implantation failure in early gestation. The follow-on reproductive studies, reproduction and
fertility, pre-implantation and post-implantation studies, implied that ADN is embryotoxic. A
mouse embryo toxicity study also showed ADN affects the embryo. The in vitro Hydra
attenuata developmental toxicity screen did not support the rodent developmental toxicity data.
EPR studies indicated that ADN can decompose to form reactive nitrogen metabolites which can
be harmful in biological systems. The hepatocyte studies suggested that ADN has the potential
for directly affecting cellular DNA in vitro. The EPR and hepatocyte results are supported by
traditional genotoxicity assays, including the Ames test, the mouse lymphoma cell mutagenesis
test, and the in vivo mouse bone marrow micronuclei assay, which all reported that ADN is
mutagenic.

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2.0 INTRODUCTION

Ammonium dinitramide (ADN) is a class 1.1 explosive oxidizer which has been considered for
use by DoD and NASA in solid rocket engine propellant mixtures and explosives (Kinkead et
al., 1995). ADN is a potential replacement for ammonium perchlorate (AP) in rocket propellant
formulations. ADN is predicted to provide better performance (between 5 and 15 percent) and a
potential 8 percent payload increase (Borman, 1994). Burning AP produces a heavy smoke trail
with high hydrochloric acid content. Chlorine is a major contributor to ozone depletion in the
stratosphere and the highly visible contrail makes rockets more vulnerable to detection and
tracking (Berty et al., 1995). ADN should not generate a heavy contrail. In 1990, the Armstrong
Laboratory, now Air Force Research Laboratory, was tasked by the USAF with assessing the
occupational and environmental risk of ADN (Steel-Goodwin et al., 1995a). Recent re-interest
in ADN as a propellant revealed that its toxicity in the occupational setting had never been
summarized in accessible literature.

ADN (NH N(NO ) ) is a light sensitive, white, water soluble powder which must be stored in
4 2 2
protective vials in an enclosed cabinet. The test compound, as commonly available through SRI
International (Menlo Park, CA), is known to be contaminated with 1 to 2 percent ammonium
nitrate (Graeter et al., 1998). ADN in aqueous solution is stable in the absence of ultraviolet light.
In the presence of UV light, ADN decomposes, accompanied by a decrease in solution pH
(Kinkead et al., 1995; Steel-Goodwin et al., 1997). Breakdown products in water include the free
radical NO-, as well as NO and NO (Steel-Goodwin et al., 1997).
2 3

3.0 TOXICITY STUDIES

Prior to the studies initiated by Armstrong Laboratory in 1990, no toxicological information was
available for this compound. Anecdotal field reports of ADN causing numbness of the fingers
indicated that the oxidizer is readily absorbed into the skin (Kinkead et al., 1995). The routes of
exposure considered most viable were dermal absorption and accidental ingestion of ADN.
Environmental exposure through drinking water contamination also constitutes a potential
exposure route. Since ADN is not volatile (calculated vapor pressure of 1.7x10-12 at 25°C)
(Clausen et al., 2007), inhalation exposure to the oxidizer would be minimal.

3.1 Acute Toxicity

ADN was tested in standard acute studies. The oral LD in young adult male F-344 rats was
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determined to be 823 mg/kg (Table 1); the mean time to death was less than one hour. This
result classifies ADN as “moderately toxic.” All deaths were preceded by convulsions; systemic
vasodilation typically caused by nitrates was also evident (Kinkead et al., 1994).

No evidence of dermal irritation or toxicity was found when ADN was tested in male New
Zealand white rabbits, the most common model for human dermal irritation. ADN was applied
at a dose of 2 g/kg topically to the backs of the rabbits (approximately 10 percent of the rabbit’s
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