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Diphenylamine and derivatives in the

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Chemosphere 53 (2003) 809

818

www.elsevier.com/locate/chemosphere

Review

Diphenylamine and derivatives in the environment: a review

Oliver Drzyzga *
Department of Marine Microbiology, Center for Environmental Research and Environmental Technology (UFT),
University of Bremen, Leobener Strasse, 28359 Bremen, Germany
Received 3 May 2002; received in revised form 31 March 2003; accepted 12 June 2003

Abstract
Diphenylamine (DPA) is a compound from the third European Union (EU) list of priority pollutants. It was assigned by the EU to Germany to assess and control its environmental risks. DPA and derivatives are most
commonly used as stabilizers in nitrocellulose-containing explosives and propellants, in the perfumery, and as
antioxidants in the rubber and elastomer industry. DPA is also widely used to prevent post-harvest deterioration
of apple and pear crops. DPA is a parent compound of many derivatives, which are used for the production of
dyes, pharmaceuticals, pho- tography chemicals and further small-scale applications. Diphenylamines are still
produced worldwide by the chemical industries. First reports showed that DPA was found in soil and groundwater.
Some ecotoxicological studies dem- onstrated the potential hazard of various diphenylamines to the aquatic
environment and to bacteria and animals. Studies on the biodegradability of DPA and its derivatives are very
sparse. Therefore, further investigation is required to determine the complete dimension of the potential
environmental hazard and to introduce possible (bio)remediation techniques for sites that are contaminated with
this class of compounds. This is the rst detailed review on DPA and some derivatives summarizing their
environmental relevance as it is published in the literature so far and this review will recommend conducting further
research in the future.
2003 Elsevier Ltd. All rights reserved.
Keywords: Diphenylamine; Nitrodiphenylamine; Aminodiphenylamine; Stabilizer; Toxicity; Microbial degradation; Bioremediation

Contents
1. Introduction.......................................................................................................................................810
2. Application and occurrence of DPA and derivatives..................................................................810
3. Chemistry and toxicity of DPA and derivatives...........................................................................811
4. Environmental fate, microbial degradation, and metabolism of DPA and derivatives............812
5. Conclusions........................................................................................................................................813
Acknowledgements...................................................................................................................................815
References...................................................................................................................................................815

Tel.: +49-421-218-7234; fax: +49-421-218-7222.

E-mail address: drzyzga@biotec.uni-bremen.de (O. Drzyzga).

0045-6535/$ - see front matter 2003 Elsevier Ltd. All rights

reserved. doi:10.1016/S0045-6535(03)00613-1

81
0

O. Drzyzga / Chemosphere 53 (2003) 809818

1. Introduction
Diphenylamine (DPA; structure see Fig. 1A) is a
chemical compound of the third European Union list
of priority pollutants. The elaboration of
recommenda- tions to assess and control the
environmental risks originating from this compound
was assigned to Ger- many by the Commission of the
European Communities (Kommission der
Europaischen Gemeinschaften, 1997). While for
aniline (the simplest primary amine of pure aromatic
structure) numerous studies concerning toxi- city and
degradability are known since many years, the
simplest secondary amine of pure aromatic structure,
DPA, has been entered into the list of priority
pollutants quite recently. Because of its antioxidative
property, DPA is a stabilizing compound predominantly
included in mono-, di-, and tri-base propellants of
missiles and nitrocellulose-containing explosives. The
mostly utilized stabilizers are diphenylamine and its
urea-substituted derivatives, the so-called acardites
and centralites (see example given in Fig. 1D).
They can be distin- guished in: acardite I
(unsymmetrical diphenyl urea), acardite II
(methyldiphenyl urea), acardite III (ethyldi- phenyl
urea); centralite I (symmetrical diethyldiphenyl urea),
centralite II (symmetrical dimethyldiphenyl urea), and
centralite III (methylethyldiphenyl urea) (Meyer,
1977). Some of these stabilizers have the additional
function to be gelatinizing compounds, an advantage
for manufacturing processes of multi-base, solventfree
gunpowders and propellants (Meyer, 1977).
Apart from military utilization, diphenylamines were
and are still used in numerous further industrial appli-

H
N

Diphenylamine

Diphenylamidogen

(A)

(B
)
CH3
NH
O

H
N

CE

Diphenylamine derivatives
(C)

Acardite II
(D)

Fig. 1. Molecular structures of diphenylamine and

derivatives. Panel 1B shows the radical structure of DPA. The
substituents AF in panel 1C are variable (see description in
the text) or covered by hydrogen atoms. Arcardite II

cations (see next section). DPA-containing stabilizers

usually have not been taken into consideration in analytical investigations of ammunition-contaminated sites.
The contamination of soils of former ammunition
plants with nitro- and amino-substituted compounds as
well as other additives (e.g., stabilizers) is a constant
source of hazard for the human health, especially if
these sub- stances pass into underground and surface
waters (Haas et al., 1990; Gorontzy et al., 1994;
Blotevogel and Gorontzy, 2000). Many of these
substances own muta- genic properties and
carcinogenic potential, which has been conrmed by
animal experiments for a large number of such
compounds (Dieter, 1994; Greim et al., 1998; Lachance
et al., 1999). It is known from nitro- aromatic
compounds that they can cause toxications in humans
connected with haemato- and hepatotoxical
alterations. Because of the multitude of substances
contained within ammunition wastes and because of
their even larger number of metabolites, a
toxicological eval- uation of the total contamination is
often very dicult to realize (Dieter, 1994; Greim et
al., 1998).

2. Application
derivatives

and

occurrence

of

DPA

and

The worldwide annual production of DPA in the

1980s was about 40 000 t of which nearly 4000 t were
produced in Germany (BUA, 1993; Rippen, 1997). The
compound still has an industrial signicance, so that
the todays annual production amount may be even
higher. This is especially supported by published values
showing high production rates for this compound in
the eastern countries of Europe (for example, the
Slovak Republic producing more than 10 000 t/year
(Murin et al., 1997)). DPA is predominantly used as
stabilizer for single- or
multi-base propellants and nitrocellulose-containing
explosives. It has the function of binding degradation
products, which develop during long-term storage (e.g.,
NO, NO2, HNO3) in order to prolong storage times.
Otherwise, the nitric degradation products would enhance further decomposition so that nally the
powder becomes useless (Espinoza, 1988; Espinoza
and Thorn- ton, 1994). During its stabilizing
reaction, DPA is
transformed to its nitrated derivatives (above all to
mono-, di- and trinitro-DPA) (Curtis and Rogasch,
1987; Espinoza, 1988; Haas et al., 1990; Espinoza and
Thornton, 1994). If DPA from ammunition waste
causes soil and water contamination, it always will be
accompanied by nitrated DPA derivatives. For example, at a

(methyldiphenyl urea) is one example for urea-substituted

derivatives of DPA (panel 1D).

O. Drzyzga / Chemosphere 53 (2003)

site of a former ammunition plant in Lower Saxony

(Germany), in which nitrocellulose had been
produced and mixed with DPA as stabilizer during
the second world war, Haas et al. (1990) could detect

809818

81
1

large fractions of contaminated material. Gunpowder

plates or discs were found at this site, which showed
high concentra- tions of DPA (1.42.9 g/kg), 2nitro-DPA (0.51.25

g/kg), and 4-nitro-DPA (0.41.2 g/kg). In another

study, Entenmann and Schacke (1994) reported on
a former military air base area in Lower Saxony
(Germany) on which a widely scattered, diuse
contamination with DPA, 2-nitro-DPA, and 4-nitroDPA in soil and in groundwater had been detected.
In general, the detection of DPA and derivatives
has not found analytical consideration in most cases
until now, although the analytical techniques are
established (see references below).
DPA is also widely used to prevent post-harvest
de- terioration (storage scald) of apple and pear crops
(Olek, 1988; Gutenmann et al., 1990; Ingle et al., 1990;
Johnson et al., 1997; Kim-Kang et al., 1998; Mir and
Beaudry, 1999). Another important application of
DPA is its use as an antioxidant for various polymers
and elastomers and as condensates for the insulation
of rubber. Some further applications include: (i) DPA
as precursor for the chemical synthesis of (azo-)dyes
such as Metanil Yellow and Orange IV (Layer and
Kehe, 1981; Shao and Young, 1994; Lye and Freeman,
1999), (ii) DPA as stabilizer in perfumery products
(Calnan, 1978; Bazin et al., 1986), (iii) its use for the
detection of oxidizers (Sugihara et al., 1993), (iv) its
use for the detection
of DNA (Decallonne and
Weyns, 1976; Gendimenico et al., 1988; Thompson
and Dvorak, 1989; Hauser and Karamata, 1992), (v) its
use as a biozid against body louse, chiggers and
housey (Layer and Kehe, 1981), and (vi) its use as a
precursor of non-steroidal anti- inammatory drugs
(Shaw et al., 1995; Masubuchi et al., 1999).
4-Amino-DPA (4A-DPA; see Fig. 1C with B
NH2) is used in the production of hair dyes and other
dyes (Singh et al., 1986, 1992), is a precursor and
intermediate for the synthesis of various chemicals
for photography and for pharmaceutical products
(Layer and Kehe, 1981; Srivastava et al., 1982; Lye and
Freeman, 1999), and is used in rubber compound
manufacture (Layer and Kehe, 1981). Additionally, 4amino-DPA is an azo reduction metabolite of the
widely used food dye Metanil Yellow (Raza et al.,
1982, 1983; Singh et al., 1986).
2-Nitro-DPA (2N-DPA; see Fig. 1C with A
NO2), 4-nitro-DPA (4N-DPA; see Fig. 1C with B
NO2), and 2,4-dinitro-DPA (24diN-DPA; see Fig. 1C
with A and B NO2) are used in dierent
dispersion colors and are components of several azo
dyes (Burkinshaw and Lu, 1993; Lye and Freeman,
1999). 4-Nitro-DPA is also used as intermediate for
the production of antioxidant ad- ditives for rubber
products (Layer and Kehe, 1981). 2-Nitro-DPA is a
component of Otto Fuel II used by the US Navy as
fuel for torpedoes and other weapon systems (Powell
et al., 1998). 2,20,4,40,6,60-Hexanitro-DPA (hexyl; see Fig.
1C with A, B, C, D, E, and F NO2) was used by
Navy military forces as underwater explosive (marine
bombs, marine mines, and torpedoes) in mix- tures
together with TNT (trinitrotoluene) and aluminium

powder and therefore represents a hazard for the

aquatic marine environment (Meyer, 1977; Urbanski,
1985; Schreiber, 1990). Nevertheless, hexyl-containing
explo- sives have not been produced anymore for
military ap- plications.
2,20,4,40-Tetrahalo-DPA (see Fig. 1C with A, B, D
and F Br or Cl) are used in biocidal reagents
(Layer and Kehe, 1981) and 2-carboxy-DPA (also
known as DPC; see Fig. 1C with A COOH) is used
as a non- specic Cl channel blocker (open pore
inhibitor) for physiological studies (Leung and Wong,
2000; Fortner et al., 2001; Fortuna et al., 2001; Kogan et
al., 2001; Trout et al., 2001).
Because of the manifold utilization of DPA and derivatives there is a lot of literature available for the
analytical detection of these compounds. Spectrophotometric, voltametric and amperometric, colorimetric
as well as numerous chromatographic methods have
been applied (TLC, HPLC, GC, and GC/MS) (Piorr
and Toth, 1967; Allen and Hall, 1980; Luke and
Cossens, 1980; Ryder and Knowlton, 1983; Curtis and
Rogasch, 1987; De Jong and Verweij, 1988; Espinoza,
1988; Olek,
1988; Ho et al., 1990; Wigeld and McLenaghan, 1990;
Bergens, 1995; Bergens and Danielsson, 1995;
Drzyzga, 1996; Drzyzga and Blotevogel, 1997; Yu et
al., 1997; Tong et al., 2001).
DPA is also described as a naturally occurring
compound in onions (Karawya et al., 1984), in leaves
of black and green tea and further plants (Nose et al.,
1971; Karawya et al., 1984; Wang et al., 2001) as well
as in the peel of citrus fruits (Piorr and Toth,
1967).
Therefore, the development of microbial
degradation strategies for this compound could have
occurred during microbial evolution.

3. Chemistry
derivatives

and toxicity of

DPA

and

DPA is a very reactive compound. This is due to

the imine hydrogen atom, which can easily be
replaced electrophilically. For example, it can be
replaced by al- kali metals, a reaction which can be
used for the detec- tion of potassium (as N
-potassium-DPA). Also metals (e.g., aluminum) are
able to displace the hydrogen under formation of
metal (e.g., aluminum) diphenylamide. Nu- merous
other derivatives are known which are formed when
the N -hydrogen is replaced by alkyl-, aryl-, or acylgroups. A transformation to carbazole can happen via
dehydrogenation of the two carbon atoms in orthoposition. The reaction with nitric compounds (as
men- tioned above) leads rapidly to N -nitroso-DPA,
followed by complete nitration to various
nitrodiphenylamines. Espinoza and Thornton (1994)
summarized some plau- sible nitrosation and
nitration mechanisms of DPA, in- cluding the classical
explanation of DPA conversion into its various
derivatives by rearrangement reactions of

N -nitrosamines to the C-nitroso compounds (the

Fisher Hepp reaction) and its subsequent oxidation
to the cor- responding nitro compounds. Also
halogenations with chlorine or bromine to produce
polyhalogenated DPA (e.g., 2,20,4,40-tetrahalo-DPA as
mentioned above) can be obtained. A large number of
further derivatives is known, which will not be listed
here. Many of these derivatives are generated from
the so-called diphenyl- amidogen, the highly
reactive radical of the deproto- nated DPA (Fig. 1B).
This unstable radical is developed during oxidation of
DPA with dierent oxidizing agents and
demonstrates its utilization property as a highly
eective anti-oxidizing agent (Sugihara et al., 1993).
For further information about the chemistry of DPA
and other diarylamines see the review of Layer and
Kehe (1981).
Little is known about the toxicity of DPA and derivatives. The current knowledge is basically limited
on the parent compound DPA and 4-amino-DPA.
DPA can enter humans orally, percutaneously or by
inhala- tion, and accidentally it leads to dermatitis
eects (formation of vesicular and exudative eczemas)
when employees worked with DPA-containing
products (Raith, 1976; Bazin et al., 1986; Hemmer et
al., 1997). Ec- zema formation, hypertension,
tachycardia, and bladder diseases have been observed
when humans had been administered alcoholic DPA
solutions (GDCh, 1988). DPA treatment in short-time
studies and during long- term expositions in animal
experiments with dogs, rats, and mice showed an
increase of organ weights, methe- moglobinemia as
well as liver, spleen, and kidney dam- aging eects
(e.g., coagulations, necroses and lesions) (Alexander
et al., 1965; Thomas et al., 1967a,b; Crocker et al., 1972;
Sorrentino et al., 1978; Kronevi and Holmberg, 1979;
Powell et al., 1983, 1984; Alvarez et al., 1987; GDCh,
1988; Mallon et al., 1988; Lenz and Carlton, 1990; Das
et al., 1992; Rohrbach et al., 1993; Lenz et al., 1995;
Masubuchi et al., 2000). The oral LD50 for rat, mouse,
and guinea pig is reported to be 1165 2000, 1750, and
300 mg/kg, respectively (RTECS, 1995; GDCh, 1988).
Yoshida et al. (1989) reported on a NOEL on male and
female F344 rats of about 111 mg/ kg/day. After oral
intake and metabolism (e.g., oxida- tion to hydroxyDPA derivatives, glucuronidase reac- tions), the
excretion takes place via urine and faeces (Das et al.,
1992). A direct mutagenic or carcinogenic eect of
DPA could not be detected in diverse Ames tests with
dierent cultures of Salmonella typhimurium (TA 98, TA
100, TA 1535, TA 1538; with or without metabolic
activation), in several DNA tests (e.g., DNA
reparation test (SOS repair system), Unscheduled
DNA Synthesis (UDS), point mutation test) as well as
in studies with rats, mice, and dogs (GDCh, 1988;
Mallon et al., 1988; Das et al., 1992; Rippen, 1997).
Ardito et al. (1996) reported on a slight increase of
sister chromatid exchanges after a 48-h inuence of
DPA on cultured

human lymphocytes. A further study indicated oxidative

DNA injuries within liver cells of rats caused by DPA
(Lodovici et al., 1997).
Contact with 4-amino-DPA leads to skin irritations
(Raza et al., 1983) and shows interaction with gut mucosal epithelium of rats causing alterations (Raza et
al., 1982). Srivastava et al. (1982) reported on binding
ca- pacity of 4A-DPA to proteins of the liver tissue
(espe- cially binding to aspartic and glutamic acid).
Binding anity of 4A-DPA to serum proteins was also
shown by Raza et al. (1983). These authors showed
the covalent binding of 4A-DPA to aspartic acid,
especially to the globulin fraction. The oral LD50 for
4A-DPA is 464 mg/ kg for rats and about 847 mg/kg
for male and female albino rats (RTECS, 1995). These
values are much lower than the LD50 value measured
for 4-nitro-DPA of above 7940 mg/kg for rats (RTECS,
1995), clearly demon- strating that the aromatic
amines in general are more toxic than the nitrated
(parent) compounds.
DPA is hazardous to organisms of the aquatic environment as shown by the EC 50 values of Kaiser and
Ribo (1988) for the bioluminescent bacterium Photobacterium phosphoreum. After 30 min contact time, a
value of about 4.75 mg/l was determined. This value
was conrmed by Drzyzga et al. (1995a) with Vibrio
scheri as model organism (~5.5 mg/l) and therefore,
the au- thors classied DPA as very toxic for aquatic
organ- isms by using a widely accepted classication
scheme. More ecotoxicological eects of various
diphenylamines are summarized in Table 1.
DPA also shows inhibitory eects on the photosynthesis of phototrophic bacteria. Goodwin (1980) reported on the inhibition of the carotenoid synthesis of
a few photosynthetic bacteria by DPA. Another study
demonstrated the negative eect of DPA on the
photo- synthetic apparatus of Rhodopseudomonas
sphaeroides. DPA treatment led to a decrease of the
spheroidin synthesis and to the accumulation of
reduced precursors (Michalski et al., 1985). Sharp et
al. (1999a,b) found that DPA is a binding QO site
inhibitor of the cytochrome bc1 of Rhodobacter
capsulatus.
Additional toxicological data of DPA are summarized in the data compilation of Rippen (1997).

4. Environmental fate, microbial degradation,

and meta- bolism of DPA and derivatives
The water solubility of a compound is decisive for
its bioavailability and eventual toxic eects.
Depending on the temperature, DPA shows a water
solubility of around 3545 mg/l (Meyer, 1977; Layer
and Kehe, 1981;
GDCh, 1988; Drzyzga, 1996; USEPA, 1998). DPA has
been classied in Germany as strongly water
endanger- ing (water endangering class 3) (Rippen,
1997). The available data indicate that DPA is stable
towards hy-

Table 1
EC values of DPA and some derivatives
Compound

EC (eective concentration) values

References

DPA

EC50 and EC100 Daphnia magna: 2.3 and 7.0 mg/l, respectively
EC50 Photobacterium phosphoreum: ~4.75 mg/l
EC50 Vibrio scheri: ~5.5 mg/l

GDCh (1988)
Kaiser and Ribo (1988)
Drzyzga et al. (1995a)

4A-DPA

EC50 Photobacterium phosphoreum: ~0.33 mg/l

EC50 Vibrio scheri: ~1.1 mg/l

Kaiser and Ribo (1988)

Drzyzga et al. (1995a)

2A-DPA

EC50 Vibrio scheri: ~1.5 mg/l

Drzyzga et al. (1995a)

4N-DPA

EC50 Vibrio scheri: ~1 mg/l

Drzyzga et al. (1995a)

2N-DPA

EC50 Vibrio scheri: ~7.7 mg/l

Drzyzga et al. (1995a)

24diN-DPA

EC50 Vibrio scheri: ~5.3 mg/l

Drzyzga et al. (1995a)

Hexyl

EC50 Vibrio scheri: ~6.3 mg/l

Drzyzga et al. (1995a)

drolysis at pH values of 5, 7, and 9 (USEPA, 1998).

DPA appears as a lipophilic substance with a distribution coecient (n-octanol/water) log Pow of 3.42 (Rippen, 1997). The vapor pressure of DPA was
determined to be around 6.39 104 Torr (USEPA,
1998). In ex- periments with the fathead minnow
(Pimephales prom- elas), a bioconcentration factor
(BCF) of 3070 was found (GDCh, 1988; Rippen,
1997). Also plants are able to enrich DPA, as for
example barley with an enrich- ment factor of 4
(Mallon
et
al.,
1988).
According
to
adsorption/desorption eects, the mobility of DPA
ranges from slightly mobile (Kads 151:57 in clay soil)
to mobile (Kads 21:43 for loamy sand, Kads 13:79
for
loam, Kads 4:92 for silt loam, and Kads 16:44 for
silty clay loam sediment (USEPA, 1998)).
The physico-chemical and biological degradation
of DPA has only been investigated to a little extend.
Thus, photolytical half-life times of DPA in water
during solar radiation of about 233 h (depending on
the season) were determined (BUA, 1993). An
irradiation of an aqueous DPA solution with a
wavelength of 313 nm at pH 7 and 25 C led to a
photolytic (oxidative) trans- formation to carbazole,
and under oxygen exclusion to a disproportionation
to carbazole and tetrahydrocarb- azole (GDCh, 1988;
Mallon et al., 1988).
The biological degradability of DPA and
substituted diphenylamines (nitrated, aminated,
methylated, halo- genated, etc.) has scarcely been
investigated until today. A former publication of
Gardner et al. (1982) refers to an aerobic metabolism
14
of C-labelled diphenylamine. After incubation of
14
C-DPA in a sewage sludge exper- iment, the
radioactivity was recovered within the me- tabolites
4-hydroxy-DPA, an unknown isomer, aniline, and
indole. These compounds were identied also in
studies on the microbial degradation and metabolism
of DPA and some derivatives under anoxic conditions
(Drzyzga, 1996; Drzyzga et al., 1996; Drzyzga and
Blotevogel, 1997). The few existing data about
products

and metabolites of the biological degradation of DPA

and derivatives are summarized in Table 2. In nearly
all studies not all metabolites have been extracted or
chromatographically identied. Gardner et al. (1982)
showed that more than 35% of the initial
14
concentration of C-labelled DPA remained as a
residual concentra- tion after 6 h of incubation in a
model sewage sludge experiment. Consequently, a
retention time of DPA- containing waste water of 26
h within the aerobic treatment complex of a sewage
plant is in any case too short, if the complete
degradation of DPA in waste water will be
guaranteed (Gardner et al., 1982).
Nevertheless, the data obtained from the available
literature allow the conclusion that a complex mixed
culture should be able to mineralize DPA and DPA
derivatives. As shown in Table 2 and in Figs. 2 and 3,
the central main product of the anaerobically,
microbi- ally mediated DPA degradation is always
aniline, a compound which is completely degradable
under oxic and anoxic conditions (e.g., Aoki et al.,
1983; Schnell and Schink, 1991; Zissi and Lyberatos,
1999; Kahng
et al., 2000). Also the aerobically,
microbially mediated DPA degradation seems to
proceed via aniline (Gardner et al., 1982) and can
therefore lead to total mineraliza- tion to CO2, NH
and H2O. In addition
to complete mineralization of
4
DPA and derivatives, side reactions leading to more
complex condensation products (e.g., carbazole,
phenazines, acridines, and benzothiazoles) have been
observed (Table 2).

5. Conclusions
DPA and derivatives still enter the environment
and these compounds represent a group of
problematic substances from the ecotoxicological
point of view. For this reason, their integration into a
controlling ana- lytic at suspicious sites with DPA
contamination is

A)

Table 2
Microbial degradation and transformation of DPA and some derivatives
Compound

Products and metabolites

References

DPA

4-Hydroxy-DPA, aniline, indole (and some unidentied metabolites

resulting from the oxidative cleavage of the aromatic ring system)
Aniline as product of DPA cleavage under anoxic conditions (by
anaerobic enrichments and sulfate-reducing bacteria)
Decrease of DPA concentration without identication of intermediates
and degradation products (DPA was used aerobically as carbon source
by three Pseudomonas species and by activated sewage sludge)
Biological and abiotic transformation of DPA to 2-OH-, 3-OH-, 4-OHand 2,40-diOH-DPA and to glycosyl conjugates of the hydroxylated
intermediates
99.6% decrease of DPA from water in an enzymatic system with
horse radish peroxidase (HRP)

Gardner et al. (1982)

Drzyzga (1996), Drzyzga and
Blotevogel (1997)
Christodoulatos et al. (1997)
Kim-Kang et al. (1998)
Verschueren (1996)

4N-DPA

4A-DPA as product of reduction under anoxic conditions (4-OH-DPA

also found)

2N-DPA

2A-DPA as product of reduction under anoxic conditions

Decrease of 2N-DPA concentration (no identication of
intermediates and degradation products; cometabolic degradation
by Clostridium species)

Drzyzga et al. (1995b, 1996)

Powell et al. (1998)

4A-DPA

Aniline as main product after cleavage; no condensation products

Drzyzga et al. (1996)

2A-DPA

Aniline as main product after cleavage; further products:

phenazine, 4-aminoacridine, indole, and methylaniline

Drzyzga et al. (1996)

24diN-DPA

2A4N-DPA as primary and 24diA-DPA as nal product of

reduction under anoxic conditions
Aniline as main product after cleavage; further products: 2aminophen- azine and three benzothiazole derivates

Hexyl

Rapid decrease of hexyl concentration (no identication of

intermediates or degradation products)

H
N

+ 2[H]

+ H2 O

HO

B)
Aniline

Benzene

Aniline

Drzyzga et al. (1995b)

Drzyzga et al. (1996)
Drzyzga (1996)

not problematic, if the DPA is totally removed when

the fruits are washed before consumption, so that it
will not pose a direct danger for the human health.
Nevertheless, the cleaning of the fruits shifts the
environmental threat of DPA and derivatives to the
wastewater side. Because of the lack of toxicity data
for diphenylamines especially with regard to human
toxicology, it becomes obvious that there is still a need
for research, particularly since these compounds have
already been detected in soil and

NH2

NH2

Drzyzga et al. (1995b, 1996)

Phenol

mineralization

Fig. 2. Microbially mediated hydrolytic cleavage (B) and reductive cleavage (A) of DPA in anaerobic enrichments and
by bacterial pure cultures (adopted and changed from
Drzyzga, 1996, 1999).

recommendable. The application of DPA to help

control apple and pear scald and to prolong their
storability is

groundwater. Various remediation techniques have already been applied for several xenobiotic compounds
and should be investigated and tested in more detail
also for DPA and its derivatives in future studies.
Aromatic amines are known to be mutagenic and
carcinogenic and their detection as well as their controlling in the environment has found an increasing
at- tention during the last years (Schmidt et al., 1998;
Rao et al., 1999, 2001). Because of the manifold
application of diphenylamines and the worldwide
diusion into the environment, the ecotoxicological
evaluation of this group of compounds should not be
ignored. As an antioxidant and therefore with the
function as a stabi- lizer, DPA will open up new elds
of application in the

NO2

H
N

NO2

H
N
O2 N

2-Nitrodiphenylamine

H
N

O2N

4-Nitrodiphenylamine

2,4-Dinitrodiphenylamine

+ 6[H] - 2 H2O
NH

+ 6[H]

- 2 H2O

+ 6[H]

- 2 H2O

H
N

O2N
2-Amino- 4-nitrodiphenylamine

- 2 H2O

+ 6[H]

NH

Alvarez, F., Diaz-Alferez, F.J., Perez-Munoz, P.J., ArteagaSerrano, F., Martin-Rodriguez, A., 1987. Cystic kidney
disease induced by diphenylamine in rats. Actas Urol.
Esp. 11, 656659.
Aoki, K., Ohtsuka, K., Shinke, R., Nishira, H., 1983. Isolation
of aniline-assimilating bacteria and physiological characterization of aniline biodegradation in Rhodococcus erythropolis AN-13. Agr. Biol. Chem. 47, 25692575.
Ardito, G., Bramanti, B., Bigatti, P., Lamberti, L., Dolara, P.,
1996.
Cytogenetic
eect
thiabendazole
and
diphenylamine
on cultured human lymphocytes: sister chromatid exchanges
and cell cycle delay. Boll. Soc. Ital. Biol. Sper. 72, 171178.
Bazin, B.H., Foussereau, J., Cavelier, C., 1986. Allergy to
diphenylamine from an industrial grease. Contact Dermatitis 14, 116.
Bergens, A., 1995. Decomposition of diphenylamine in nitro-

NH

H
N

H
N

H2N

H2N
2-Aminodiphenylamine

H
N

4-Aminodiphenylamine

+ 2[H]

2,4-Diaminodiphenylamine

+ 4[H]
(- NH3)

+ 2[H]
NH2

2 Aniline
mineralization

condensation reactions

Fig.
3.
Microbially
mediated
reduction
of
nitrodiphenylamines and subsequent reductive cleavage of
the intermediates to ani- line in anaerobic enrichments and
bacterial pure cultures. The formation of 1,3diaminobenzene was never observed and the release of
ammonia is hypothetical, but was not determined (adopted
and changed from Drzyzga, 1996, 1999).

future and eventually increasingly develop to a chemical

more enriching in the environment.

Acknowledgements
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