Evidence for a novel heme adduct generated by the in vitro reaction of 2,4,6-trinitrotoluene with human hemoglobin using electrospray ionization mass spectrometry

doi:10.1016/S0162-0134(97)00108-6

Journal of Inorganic Biochemistry

Volume 68, Issue 4, December 1997, Pages 273-278

https://doi.org/10.1016/S0162-0134(97)00108-6 Get rights and content

Abstract

The bioactivation of nitroaromatic compounds to highly reactive intermediates is responsible for the genotoxic and cytotoxic effects by reaction with DNA and proteins. Due to its continued use as a secondary explosive and its prevalence at contaminated sites, the mechanism of covalent binding of 2,4,6-trinitrotoluene (TNT), or its metabolites, to critical cellular proteins has been of interest. Herein, we report the in vitro reaction of TNT with human hemoglobin under anaerobic and reductive (using sodium bydrosulfite) conditions, yielding a novel adduct between a putative nitrosodinitrotoluene (MW = 211 Da) and the prosthetic beme group (iron protoporpbyrin-IX or beme b). While the covalent modification of hemoglobin polypeptide chains by TNT has been established, to our knowledge, this is the first example of a beme-TNT related adduct. This finding could be of relevance in investigation of biotransformation of TNT in subjects exposed to TNT via skin exposure or inhalation.

References (44)

J. Yinon et al.
J. Chromatogr.
(1985)
W.J. Buttner et al.
Analytica Cbimica Acta
(1997)
Y.-Y. Liu et al.
Chem. Biol. Interactions
(1992)
K.H. Leung et al.
Chem. Biol. Interactions
(1995)
Y.-Y. Liu et al.
Toxicol. Letters
(1995)
Y.-T. Li et al.
J. Am. Soc. Mass Spectrum.
(1993)
K.H. Leung et al.
Chem. Biol. Interactions
(1995)
G. Sabbioni et al.
J. Chromatogr.
(1996)
J.S. Olson
Methods in Enzymology
(1981)
P.R. Ortiz de Montellano
Biochimie
(1995)

L.C. Shrives-Lake et al.

Environ. Sci. Technol.

(1997)

A.Z. Li et al.

Environ. Sci. Technol.

(1997)

M. Vanderford et al.

Biotechnology Lett.

(1997)

B.H. Woolen et al.

British J. Industrial Medicine

(1986)

G. Sabbioni

Environ. Health Perspect.

(1994)

G. Sabbioni

Chem. Res. Toxicol.

(1994)

I. Zwimer-Baier et al.

Environ. Health Perspect.

(1994)

P.M. Wilson et al.

Arch. Toxicol.

(1996)

P. Eyer

Environ. Health Perspect.

(1994)

H. Savolainen et al.

British J. Industrial Medicine

(1985)

A. Zitting et al.

Arch. Toxicol.

(1982)

L.W. Chang et al.

Annu. Rev. Pharmacol. Toxicol.

(1994)

Cited by (10)

Accumulation of trinitrotoluene (TNT) in aquatic organisms: Part 1 - Bioconcentration and distribution in channel catfish (Ictalurus punctatus)
2005, Chemosphere
Little is currently known regarding the toxicokinetics of TNT in fish. In the present study, the bioconcentration and distribution of trinitrotoluene (TNT) and TNT biotransformation products was investigated in juvenile channel catfish by exposing catfish to ¹⁴C-labeled TNT in water. Uptake experiments showed relatively fast rates (k_u = 10.1 ml g⁻¹ h⁻¹) for TNT from the water; however, bioconcentration factors for TNT were low (0.79 ml g⁻¹) due to rapid biotransformation and potential elimination of TNT. Accumulation of extractable radioactivity (TNT and all extractable biotransformation products) was much greater (BCF = 10.5 ml g⁻¹) than that for parent compound. TNT (parent compound) bioconcentrated to the greatest extent in the gills of the fish, while total radioactivity bioconcentrated to the greatest extent in the viscera. Residual portions of the fish that contained muscle and skin had lower concentrations of TNT than the whole fish, indicating that ingestion of fish filets would result in decreased exposure to human consumers. Although the bioconcentration potential of TNT is very low, future research needs to be conducted to identify the biotransformation products that make up most of the radioactivity in exposed fish and evaluate their potential to promote toxicity.
Preliminary kinetics and metabolism of 2,4,6-trinitrotoluene and its reduced metabolites in an aquatic oligochaete
2004, Aquatic Toxicology
We examined the toxicokinetics and metabolism of 2,4,6-trinitirotoluene (TNT) and four of its major reduced metabolites (2-amino-4,6-dinitrotoluene (2ADNT), 4-amino-2,6-dinitrotoluene (4ADNT), and 2,4-diamino-6-nitrotoluene (2,4DANT)) in the freshwater, aquatic oligochaete Tubifex tubifex exposed to spiked, reconstituted water. In uptake experiments with each compound, steady state concentrations were reached within 1 h, and all absorbed compounds were completely eliminated in 0–3 h. The appearance of 2ADNT and 4ADNT (from metabolism of absorbed TNT) was five times slower, reaching 95% of steady state in 14.2–16.1 h. Approximately, 82% of absorbed TNT was metabolized to ADNTs; metabolism to 4ADNT was favored over 2ADNT by a factor of 3. No further metabolism of ADNTs to DANTs was detected. After a loss of 29–50% of metabolically-generated ADNTs during the first hour of the TNT depuration experiment, Tubifex ADNT concentrations remained constant throughout the 53 h depuration period. This suggests differences between the toxicokinetics of absorbed ADNTs and the toxicokinetics of metabolically-generated ADNTs. Experiments using radiolabeled ( $^{14} C$ ) TNT revealed that only 15–47% of $^{14} C$ -TNT equivalents were identified as TNT, 2ADNT, or 4ADNT, indicating significant metabolism and/or binding to biomolecules. Of unidentified $^{14} C$ -TNT equivalents, 28–38% remained unextractable. Both unextractable and extractable unidentified substances increased throughout the 54 h $^{14} C$ -TNT uptake experiment. The unidentified portions of the radiolabel were not eliminated during a 53-h depuration experiment. Bioconcentration factors (BCFs) for HPLC-detectable compounds were found to be linearly related to log K_OW (r² = 0.9963). BCFs for 2ADNT, 4ADNT, and 2,4DANT were 10.22, 12.41, and 2.75, respectively. The BCF for TNT was 12.25, based on a molar sum of total TNT and its metabolites (∑TNT + 2ADNT + 4ADNT), and 2.53 based on TNT only. Compound hydrophobicity predicted the toxicokinetics and bioconcentration of compounds absorbed from water, however, the large discrepancy between the toxicokinetics of absorbed versus metabolically-generated ADNTs and the bioconcentration and toxicokinetics of the unidentified substances warrant further investigation.
Bioremediation of soils contaminated with explosives
2004, Journal of Environmental Management
The large-scale industrial production and processing of munitions such as 2,4,6-trinitrotoluene (TNT) over the past 100 years led to the disposal of wastes containing explosives and nitrated organic by-products into the environment. In the US, the Army alone has estimated that over 1.2 million tons of soil have been contaminated with explosives, and the impact of explosives contamination in other countries is of similar magnitude. In recent years, growing concern about the health and ecological threats posed by man-made chemicals have led to studies of the toxicology of explosives, which have identified toxic and mutagenic effects of the common military explosives and their transformation products (Bruns-Nagel et al., 1999a, Fuchs et al., 2001, Homma-Takeda et al., 2002, Honeycutt et al., 1996, Rosenblatt et al., 1991, Spanggord et al., 1982, Tan et al., 1992, Won et al., 1976). Because the cleanup of areas contaminated by explosives is now mandated because of public health concerns, considerable effort has been invested in finding economical remediation technologies. Biological treatment processes are often considered, since these are usually the least expensive means of destroying organic pollution. This review examines the most important groups of chemicals that must be treated at sites contaminated by explosives processing, the chemical and biological transformations they undergo, and commercial processes developed to exploit these transformations for treatment of contaminated soil. We critically examine about 150 papers on the topic, including approximately 60 published within the past 5 years.
Bioremediation of 2,4,6-Trinitrotoluene explosive residues
2014, Environmental Science and Engineering (Subseries: Environmental Science)
Ion spectrometric detection technologies for ultra-traces of explosives: A review
2011, Mass Spectrometry Reviews
Bioconcentration, bioaccumulation, and biotransformation of explosives and related compounds in aquatic organisms
2009, Ecotoxicology of Explosives

View all citing articles on Scopus

View full text

Evidence for a novel heme adduct generated by the in vitro reaction of 2,4,6-trinitrotoluene with human hemoglobin using electrospray ionization mass spectrometry

Abstract

J. Chromatogr.

Analytica Cbimica Acta

Chem. Biol. Interactions

Chem. Biol. Interactions

Toxicol. Letters

J. Am. Soc. Mass Spectrum.

Chem. Biol. Interactions

J. Chromatogr.

Methods in Enzymology

Biochimie

Environ. Sci. Technol.

Environ. Sci. Technol.

Biotechnology Lett.

British J. Industrial Medicine

Environ. Health Perspect.

Chem. Res. Toxicol.

Environ. Health Perspect.

Arch. Toxicol.

Environ. Health Perspect.

British J. Industrial Medicine

Arch. Toxicol.

Annu. Rev. Pharmacol. Toxicol.