Nitrobenzene

General information
Key Points
Fire

Flammable
Violently reacts with strong oxidants, acids and nitrogen oxides
Emits toxic fumes of nitrogen oxides when heated to decomposition
In the event of a fire involving nitrobenzene, use fine water and liquid tight chemical
protective clothing with breathing apparatus

Health

Toxic by inhalation, ingestion and skin absorption

Possibly carcinogenic in humans
May cause reproductive toxicity
The onset of symptoms may be delayed 1-4 hours after exposure to nitrobenzene
Inhalation can cause irritation of the respiratory tract, nausea, headache, dizziness,
shortness of breath and in extreme cases could lead to coma and death
Ingestion of nitrobenzene may cause gastrointestinal irritation with nausea, vomiting
and diarrhoea, as well as symptoms similar to those for inhalation
Inhalation and ingestion may also cause vertigo and bluish colouration of the skin due
to a condition called methaemoglobinaemia, with drowsiness, high blood pressure,
convulsions, anaemia, jaundice and kidney failure
Skin contact with nitrobenzene may result in mild skin irritation and eye contact may
lead to mild eye irritation

Environment

Dangerous for the environment

Inform Environment Agency of substantial incidents

Prepared by J C Wakefield
CHAPD HQ, HPA
2008
Version 1

NITROBENZENE GENERAL INFORMATION

Background
Nitrobenzene is a colourless or yellow oily
liquid, with an odour of bitter almonds and is
flammable following moderate heating.
Nitrobenzene does not occur naturally and is
therefore only produced synthetically for
industrial applications. One of the major uses
for nitrobenzene is for the production of
aniline, which is a chemical intermediate
used
during
the
manufacture
of
polyurethane. Nitrobenzene is also used
industrially in the manufacture of some
pharmaceuticals, dyes and rubbers, as a
constituent in some polishes and paint
solvents and as a solvent in the refining of
petroleum.

nitrobenzene may cause jaundice (yellow

discolouration of the skin), failure of the
kidneys, coma and even death.

Contact of the skin or eyes with vapours or

liquid nitrobenzene can cause irritation at the
site of contact. Nitrobenzene can also be
absorbed through the skin and contact with
large quantities could result in adverse
effects similar to those seen following
inhalation or ingestion.
Repeated exposure to nitrobenzene by any
route may cause damage to the liver in
addition to the effects resulting from a single
exposure.

Exposure of the general public to

nitrobenzene is extremely unlikely as it is not
commonly used in the home in substantial
quantities. The most common source of
exposure to considerable amounts of
nitrobenzene is in the workplace, either
where it is produced, or during the
production of other materials.
Nitrobenzene is very toxic and irritant to
humans who are exposed to it either by
inhalation, ingestion or skin contact.
Exposure to vapours or splashes of
nitrobenzene will cause irritation of the skin,
eyes and respiratory tract. The main effect of
inhaling or ingesting nitrobenzene is a
condition
called
methaemoglobinaemia,
which affects the ability of the blood to carry
oxygen. The effect of this includes
headache, nausea, vomiting, weakness,
dizziness, vertigo, bluish discolouration of
the skin, rapid heart rate and breathlessness.
These symptoms may be delayed for up 1 to
4 hours following exposure. In some cases,

Children exposed to nitrobenzene are

expected to show similar adverse health
effects to those seen in exposed adults,
although the effects are expected to be more
severe, as children are particularly
susceptible to the adverse effects of
exposure to nitrobenzene. There is evidence
to suggest that exposure to nitrobenzene
during pregnancy, at concentrations that may
harm the mother, may possibly cause
adverse effects to the unborn child.
Nitrobenzene has been classified as possibly
having the ability to cause cancer in humans,
by the International Agency for Research on
Cancer.

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NITROBENZENE GENERAL INFORMATION

Production and Uses

Key Points

Nitrobenzene does not occur naturally in the environment

It is produced industrially by reacting benzene with sulphuric acid, nitric acid and
water
Nitrobenzene is used in the production of aniline, a chemical used to manufacture
polyurethane foams
Nitrobenzene is also used to manufacture pharmaceuticals, rubbers, dyes, polishes
and paint solvents

Nitrobenzene does not occur naturally in the environment. It is produced industrially by

reacting benzene with sulphuric acid, nitric acid and water.
One of the major uses for nitrobenzene is for the production of aniline, which is a chemical
intermediate used during the manufacture of polyurethane foams. Nitrobenzene is also used
industrially in the manufacture of some dyes, rubbers and pharmaceuticals (it is an important
component in the production of paracetamol). Nitrobenzene is also used as a constituent in
some polishes and paint solvents and as a solvent in the refining of petroleum.
Historically nitrobenzene was used in perfumes and as a flavouring known as artificial oil of
bitter almonds due to its strong odour. This use of nitrobenzene has long been discontinued
due to its toxicity.

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NITROBENZENE GENERAL INFORMATION

Frequently Asked Questions

What is nitrobenzene?
Nitrobenzene is a colourless or yellow oily liquid, with an odour of bitter almonds and is
flammable following moderate heating. The main use of nitrobenzene is in the chemical
synthesis of aniline, which is a chemical used in the manufacture of polyurethane foams.
How does nitrobenzene get into the environment?
Nitrobenzene does not occur naturally in the environment and as such is most likely to enter
the environment from workplaces where it is manufactured or used.
How will I be exposed to nitrobenzene?
As nitrobenzene is only used industrially, it is unlikely that you will be exposed to significant
amounts unless you work with it. Nitrobenzene will form a vapour at room temperature, and
therefore anyone working with it is at risk of inhaling the vapours if adequate protective
equipment is not used. People working with nitrobenzene are also at risk of getting splashes
of it on their skin if they are not adequately protected.
If there is nitrobenzene in the environment will I have any adverse health effects?
The presence of nitrobenzene in the environment does not always lead to exposure. Clearly,
in order for it to cause any adverse health effects you must come into contact with it. You
may be exposed by breathing, eating, or drinking the substance or by skin contact. Following
exposure to any chemical, the adverse health effects you may encounter depend on several
factors, including the amount to which you are exposed (dose), the way you are exposed, the
duration of exposure, the form of the chemical and if you were exposed to any other
chemicals.
Nitrobenzene is an irritant and exposure to vapours or splashes will cause irritation of the
skin, eyes and respiratory tract. Nitrobenzene is toxic by inhalation, ingestion and skin
contact. The main symptom of exposure to nitrobenzene is a condition called
methaemoglobinaemia, which affects the ability of the blood to carry oxygen. This condition
is associated with headache, nausea, vomiting, weakness, dizziness, vertigo, bluish
discolouration of the skin, rapid heart rate and breathlessness. The symptoms may be
delayed for up 1 to 4 hours following exposure. In some cases, nitrobenzene may cause
jaundice (yellow discolouration of the skin), failure of the kidneys, coma and even death.
Can nitrobenzene cause cancer?
Nitrobenzene has been classified by the International Agency for Research on Cancer as
possibly causing cancer in humans.
Does nitrobenzene affect children or damage the unborn child?
Children exposed to nitrobenzene are expected to show similar adverse health effects to
those seen in exposed adults, although the effects are expected to be more severe, as
children are particularly susceptible to the adverse effects of exposure to nitrobenzene.
There is evidence to suggest that exposure to nitrobenzene during pregnancy, at
concentrations that may harm the mother, may possibly cause adverse effects to the unborn
child.

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NITROBENZENE GENERAL INFORMATION

What should I do if I am exposed to nitrobenzene?

It is very unlikely that the general population will be exposed to a level of nitrobenzene high
enough to cause adverse health effects.

This document from the HPA Centre for Radiation, Chemical and Environmental Hazards reflects
understanding and evaluation of the current scientific evidence as presented and referenced in this
document.

General information: Page 5 of 5

Nitrobenzene
Incident management
Key Points
Fire

Flammable
Violently reacts with strong oxidants, acids and nitrogen oxides
Emits toxic fumes of nitrogen oxides when heated to decomposition
In the event of a fire involving nitrobenzene, use fine water and liquid tight chemical
protective clothing with breathing apparatus

Health
Toxic by inhalation, ingestion and dermal absorption
The onset of symptoms may be delayed 1-4 hours after exposure to nitrobenzene
Inhalation can cause irritation of the respiratory tract, nausea, headache, dizziness,
shortness of breath and in extreme cases could lead to coma and death
Ingestion of nitrobenzene may cause gastrointestinal irritation with nausea, vomiting
and diarrhoea, as well as symptoms similar to those for inhalation
Inhalation and ingestion may also cause vertigo and cyanosis due to
methaemoglobinaemia with drowsiness, hypotension, convulsions, haemolytic
anaemia, jaundice and renal failure
Dermal exposure to nitrobenzene may result in mild skin irritation and ocular
exposure may lead to mild eye irritation
Environment
Dangerous for the environment
Inform Environment Agency of substantial incidents

CRCE HQ, HPA

03/2012
Version 2

NITROBENZENE INCIDENT MANAGEMENT

Hazard Identification
Standard (UK) Dangerous Goods Emergency Action Codes(a)
UN

1662

EAC

2X

APP

Class

6.1

Sub
risks

Nitrobenzene
Use fine water spray. Wear liquid-tight chemical protective
clothing in combination with breathing apparatus*. Spillages
and decontamination run-off should be prevented from
entering drains and watercourses.

Toxic substance

Hazards

HIN

60

Toxic or slightly toxic substance

UN United Nations number; EAC Emergency Action Code; APP Additional Personal
Protection; HIN - Hazard Identification Number
* Liquid-tight chemical protective clothing (BS 8428) in combination with self-contained open
circuit positive pressure compressed air breathing apparatus (BS EN 137).

Dangerous Goods Emergency Action Code List 2011. National Chemical Emergency Centre
(NCEC). The Stationary Office, London.

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NITROBENZENE INCIDENT MANAGEMENT

Chemical Hazard Information and Packaging for Supply Classification(a)

Carc. cat 3

Category 3 carcinogen

Repr. Cat 3

Category 3 reproductive toxin

Classification
T

Toxic

Dangerous for the environment

R23/24/25

Limited evidence of a carcinogenic effect

R40
Risk phrases

R48/23/24
R51/53

Safety phrases

Toxic by inhalation, in contact with skin and if swallowed

Toxic: danger of serious damage to health by prolonged
exposure through inhalation and in contact with skin
Toxic to aquatic organisms, may cause long-term adverse
effects in the aquatic environment

R62

Possible risk of impaired fertility

S1/2

Keep locked up and out of the reach of children

S28

After contact with skin, wash immediately with plenty of ... (to
be specified by the manufacturer)

S36/37
S45
S61

Wear suitable protective clothing and gloves

In case of accident or if you feel unwell seek medical advice
immediately (show the label where possible)
Avoid release to the environment. Refer to special
instructions/safety data sheet

Annex VI to Regulation (EC) No 1272/2008 on Classification, Labelling and Packaging of

Substances and Mixtures- Table 3.2.
http://esis.jrc.ec.europa.eu/index.php?PGM=cla (accessed 03/2012)

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NITROBENZENE INCIDENT MANAGEMENT

Globally Harmonised System of Classification and Labelling of Chemicals

)
(GHS)(a

Hazard Class
and Category

Hazard
Statement

Signal Words

Carc. 2

Carcinogenicity, category 2

Repr. 2

Toxic to reproduction, category 2

Acute Tox. 3

Acute toxicity (inhalation, dermal and

oral), category 3

STOT RE 1

Specific target organ systemic toxicity

following repeated exposure, category 1

Aquatic
Chronic 2

Chronic hazard to the aquatic

environment, category 2

H351

Suspected of causing cancer

H361f

Suspected of damaging fertility

H331

Toxic if inhaled

H311

Toxic in contact with skin

H301

Toxic if swallowed

H372

Causes damage to organs through prolonged or repeated

exposure

H411

Toxic to aquatic life with long lasting effects

DANGER

Implemented in the EU on 20 January 2009.

a

Annex VI to Regulation (EC) No 1272/2008 on Classification, Labelling and Packaging of

Substances and Mixtures- Table 3.1.
http://esis.jrc.ec.europa.eu/index.php?PGM=cla (accessed 03/2012)

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NITROBENZENE INCIDENT MANAGEMENT

Physicochemical Properties
CAS number

98-95-3

Molecular weight

123

Empirical formula

C6H5NO2

Common synonyms

Nitrobenzol

State at room temperature

Liquid

Volatility

Vapour pressure 0.3 mm Hg at 25C

Specific gravity

1.2 at 20C (water = 1)

Flammability

Flammable, after moderate heating

Lower explosive limit

1.8%

Upper explosive limit

40.0%

Water solubility

Low solubility in water, 1.8g L-1 at 25C. Soluble in ethanol,

acetone and benzene

Reactivity

Reactive. Nitrobenzene reacts violently with strong oxidants,

strong acids and nitrogen oxides, causing risk of fire and
explosion

Reaction or degradation
products

Releases toxic fumes of nitrogen oxides when heated to

decomposition

Odour

Bitter almonds

Structure

References(a,b,c)

Nitrobenzene (HAZARDTEXT Hazard Management). In: Klasco RK (Ed): TOMES System,

Thomson Micromedex, Greenwood Village, Colorado, USA. (electronic version). RightAnswer.com,
Inc., Midland, MI, USA, Available at: http://www.rightanswerknowledge.com/data/dt/dt193.htm
(accessed 01/2012).
b
The Merck Index (14th Edition). Entry 6588: Nitrobenzene, 2006.
c
The Dictionary of Substances and their Effects. Ed. S Gangolli. Second Edition, Volume 5, 1999.
a

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NITROBENZENE INCIDENT MANAGEMENT

Threshold Toxicity Values

EXPOSURE VIA INGESTION
g
4.3 -11

SIGNS AND SYMPTOMS

Unconsciousness, cyanosis, circulatory collapse,
rapid and shallow breathing and tachycardia

REFERENCES
a

International Programme on Chemical Safety, Environmental Health Criteria 230: Nitrobenzene,

2003.

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NITROBENZENE INCIDENT MANAGEMENT

Published Emergency Response Guidelines

Emergency Response Planning Guideline (ERPG) Values
Listed value
(ppm)
ERPG-1*
ERPG-2**
ERPG-3***

Calculated value
(mg m-3)

Data not available

* Maximum airborne concentration below which it is believed that nearly all individuals could be
exposed for up to 1 hr without experiencing other than mild transient adverse health effects or
perceiving a clearly defined, objectionable odour.
** Maximum airborne concentration below which it is believed that nearly all individuals could be
exposed for up to 1 hr without experiencing or developing irreversible or other serious health effects or
symptoms which could impair an individual's ability to take protective action.
*** Maximum airborne concentration below which it is believed that nearly all individuals could be
exposed for up to 1 hr without experiencing or developing life-threatening health effects.

Acute Exposure Guideline Levels (AEGLs)

10 min

30 min

ppm
60 min

4 hr

8 hr

AEGL-1
AEGL-2
AEGL-3

Data not available

The level of the chemical in air at or above which the general population could experience notable
discomfort.

The level of the chemical in air at or above which there may be irreversible or other serious longlasting effects or impaired ability to escape.

The level of the chemical in air at or above which the general population could experience lifethreatening health effects or death.

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NITROBENZENE INCIDENT MANAGEMENT

Exposure Standards, Guidelines or Regulations

Occupational Standards
LTEL (8 hour reference period): 0.2 ppm
(1 mg m-3)

WEL(a)
http://www.hse.gov.uk/

STEL(15 min reference period): No guideline value

specified

Public Health Guidelines

DRINKING
WATER QUALITY GUIDELINE

No guideline value specified

AIR QUALITY GUIDELINE

No guideline value specified

SOIL GUIDELINE VALUE AND

HEALTH CRITERIA VALUES

No guideline value specified

WEL Workplace exposure limit; LTEL - Long-term exposure limit; STEL Short-term
exposure limit

EH40/2005 Workplace Exposure Limits (second edition, published 2011).

http://www.hse.gov.uk/pubns/priced/eh40.pdf (accessed 01/2012)

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NITROBENZENE INCIDENT MANAGEMENT

Health Effects
Major Route of Exposure(a)

Toxic by inhalation, ingestion and dermal absorption

Immediate signs or symptoms of acute exposure(a,b,c)

After ingestion, inhalation and skin exposure the onset of symptoms may be delayed
1-4 hours. Symptoms include headache, vertigo and cyanosis due to
methaemoglobinaemia with drowsiness, shallow respiration, hypotension,
convulsions, haemolytic anaemia, jaundice and renal failure.
Inhalation causes irritation of the respiratory tract. Exposure to large amounts of
nitrobenzene can cause nausea, headache, dizziness, shortness of breath and in
extreme cases coma and death
Ingestion causes gastrointestinal irritation with nausea, vomiting and diarrhoea.
Ingestion of high levels of nitrobenzene can cause symptoms similar to those after
inhalation
Dermal exposure can cause skin irritation
Nitrobenzene can be irritating to the eyes causing an immediate stinging and burning
sensation with lacrimation

TOXBASE - http://www.toxbase.org (accessed 01/2012)

a
TOXBASE: Nitrobenzene, 2007.
b
TOXBASE: Nitrobenzene features and management, 2007.
c
TOXBASE: Eye irritants, 2002.

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NITROBENZENE INCIDENT MANAGEMENT

Decontamination and First Aid

Important Notes

Ambulance staff, paramedics and emergency department staff treating chemicallycontaminated casualties should be equipped with Department of Health approved,
gas-tight (Respirex) decontamination suits based on EN466:1995, EN12941:1998
and prEN943-1:2001, where appropriate.
Decontamination should be performed using local protocols in designated areas such
as a decontamination cubicle with adequate ventilation.

Dermal Exposure(a)

Remove patient from exposure.

The patient should remove all clothing and personal effects.
Double-bag soiled clothing and place in a sealed container clearly labelled as a
chemical hazard.
Gently blot away any adherent liquid from the patient.
Wash hair and all contaminated skin with copious amounts of water (preferably warm)
and soap for at least 10-15 minutes. Decontaminate open wounds first and avoid
contamination of unexposed skin.
Pay special attention to skin folds, axillae, ears, fingernails, genital areas and feet.

Ocular Exposure(b)

Remove patient from exposure.

Remove contact lenses if present and immediately irrigate the affected eye
thoroughly with water or 0.9% saline for at least 10-15 minutes.
Patients with corneal damage or those whose symptoms do not resolve rapidly
should be referred for urgent ophthalmological assessment.

Inhalation(c)

Remove patient from exposure.

Ensure a clear airway and adequate ventilation.
Give oxygen to symptomatic patients.
Apply other supportive measures as indicated by the patients clinical condition.

Ingestion(c)

Ensure a clear airway and adequate ventilation.

Give oxygen to symptomatic patients.
Apply other supportive measures as indicated by the patients clinical condition.

This document from the HPA Centre for Radiation, Chemical and Environmental Hazards reflects
understanding and evaluation of the current scientific evidence as presented and referenced in this
document.
TOXBASE - http://www.toxbase.org (accessed 01/2012)
a
TOXBASE: Nitrobenzene features and management, 2007.
b
TOXBASE: Eye irritants, 2002.
c
TOXBASE: Nitrobenzene, 2007.

Incident management: Page 10 of 10

Nitrobenzene
Toxicological overview
Key Points
Kinetics and metabolism

Nitrobenzene is readily absorbed following inhalation, ingestion and dermal contact

Dermal contact with nitrobenzene may give rise to systemic toxicity due to extensive
dermal absorption
Nitrobenzene is slowly excreted in the urine and faeces either unchanged or as the
major metabolites p-aminophenol and p-nitrophenol

Health effects of acute exposure

Acute exposure to nitrobenzene may cause methaemoglobinaemia, with symptoms

including headache, nausea, dizziness, fatigue, shortness of breath, cyanosis and
convulsions
The onset of methaemoglobinaemia may be delayed for 1 to 4 hours post exposure
Severe acute exposure to nitrobenzene can cause jaundice, renal failure, coma and
may be fatal
Dermal or ocular contact with nitrobenzene may cause mild irritation

Health effects of chronic exposure

Chronic exposure to nitrobenzene can cause methaemoglobinaemia similar to that

seen following acute exposure
Chronic nitrobenzene exposure may give rise to headache, nausea, vertigo,
confusion, hyperalgesia, haemolytic anaemia and toxic hepatitis
Nitrobenzene may cause adverse effects on male fertility due to testicular atrophy
Nitrobenzene is considered to be a possible human carcinogen

Prepared by J C Wakefield
CHAPD HQ, HPA
2008
Version 1

NITROBENZENE TOXICOLOGICAL OVERVIEW

Toxicological Overview
Summary of Health Effects
Nitrobenzene is readily absorbed following inhalation, ingestion and dermal contact [1].
There is no human information available regarding oral ingestion of nitrobenzene, however,
its absorption is expected to be extensive. Nitrobenzene can undergo extensive dermal
absorption, therefore dermal contact with nitrobenzene may cause considerable systemic
toxicity [1, 2].
The principal characteristic health effect following acute exposure to nitrobenzene is the
development of methaemoglobinaemia, which can give rise to symptoms including
headache, nausea, dizziness, drowsiness, shortness of breath, fatigue, cyanosis and
convulsions [1, 3, 4]. The onset of methaemoglobinaemia may be delayed for 1 to 4 hours
post exposure [4]. In some cases of acute nitrobenzene ingestion haemolytic anaemia may
develop approximately 5 days post-exposure [1]. Serious acute exposure to nitrobenzene
may result in jaundice, liver failure, renal failure, coma and can potentially be fatal [1-3, 5].
Dermal or ocular exposure to vapours or liquid splashes of nitrobenzene may cause mild
irritation [4].
Long term occupational exposure to nitrobenzene is associated with similar adverse effects
as those observed following a large single acute exposure, such as the development of
methaemoglobinaemia [1, 3]. Some symptoms related to chronic nitrobenzene exposure may
include headache, nausea, vertigo, confusion and hyperalgesia. Chronic exposure to
nitrobenzene may also give rise to the development of haemolytic anaemia and toxic
hepatitis [1, 5].
There is evidence from animal studies that nitrobenzene exposure may cause adverse
effects on male fertility due to testicular atrophy [1, 3, 5]. Studies in female animals suggest
that nitrobenzene does not cause embryotoxicity or teratogenicity at concentrations which do
not cause significant maternal toxicity [1]. However, no studies were located regarding
reproductive or developmental effects of nitrobenzene following chronic exposure in humans.
The International Agency for Research on Cancer (IARC) have evaluated that there is
insufficient evidence for the carcinogenicity of nitrobenzene in humans. However, there is
sufficient evidence that nitrobenzene is carcinogenic in experimental animals. Therefore,
based on the animal carcinogenicity data, nitrobenzene has been classified overall as
possibly carcinogenic to humans (group 2B) [5].

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Kinetics and metabolism

Nitrobenzene is readily absorbed following inhalation, ingestion and dermal contact [1].
Following a 6-hour inhalation exposure of human volunteers to nitrobenzene the pulmonary
absorption was found to be extensive, with uptake in the range of 73-87%. No human studies
have been located regarding oral exposure of humans to nitrobenzene, although based on
the effects of accidental poisonings, oral absorption is expected to be rapid and extensive.
Animal studies have shown significant absorption from the gastrointestinal tract following oral
exposure to nitrobenzene [1, 3]. The dermal absorption of nitrobenzene has not been well
studied, in either humans or experimental animals. However, there have been numerous
incidences of human poisonings following dermal exposure to nitrobenzene, suggesting that
dermal absorption is also of considerable importance [1, 2].
Studies involving oral administration of nitrobenzene to rabbits demonstrated distribution to
the blood, liver, kidney and lung, and in rats, nitrobenzene was detected in all tissues,
excluding stomach and intestines, particularly in kidney and intestinal fat and skeletal
muscle. Following accidental nitrobenzene poisoning in humans, the highest concentration
was found in the liver, brain, blood and stomach [1]. No studies were located regarding the
distribution of nitrobenzene following inhalation or dermal exposure.
There are few studies investigating the metabolism of nitrobenzene in humans. Guinea pigs
administered nitrobenzene intraperitoneally showed the main metabolite was p-nitrophenol
[1].
The main route of excretion of nitrobenzene is in the urine and to a lesser extent in the
faeces. In rats administered nitrobenzene as a single oral dose, approximately 65% of the
applied dose had been excreted in the urine and 15 % in the faeces by 5 days post-exposure
either as metabolites or unchanged [1, 3].
In cases of human toxicity following either dermal or inhalation exposure to nitrobenzene, the
major urinary metabolites identified were p-aminophenol and p-nitrophenol [3]. In a human
volunteer study following the oral administration of 30 mg nitrobenzene the urinary excretion
of p-nitrophenol was slow, with an initial elimination half-life of approximately 5 hours,
followed by a secondary slow phase with an elimination half-life of over 20 hours [1]. The
presence of p-aminophenol in urine has been used in biomonitoring studies as a marker of
occupational exposure to nitrobenzene [1]. Following exposure to nitrobenzene vapours, any
absorbed material was excreted in the urine as p-nitrophenol. Similarly, urinary excretion of
p-nitrophenol was reported in individuals following inhalation of nitrobenzene [1].

Sources and route of human exposure

The major source of exposure to nitrobenzene is from occupational exposure, since its
principal use is in industrial processes, such as during the production of aniline [1, 3].
Nitrobenzene is unlikely to be present in significant quantities in domestic situations and
does not occur naturally but can be formed at low levels by the reaction of benzene and
nitrogen oxides in the atmosphere [1]. In the occupational context, in addition to controlling
airborne levels to the occupational exposure standard it is also necessary to protect against
skin absorption by the use of appropriate protective clothing [1, 2].
The major routes of occupational exposure to nitrobenzene are by inhalation of vapours or
dermal absorption. Nitrobenzene is a considerable occupational exposure hazard as it
readily forms a vapour at room temperature and also may undergo significant dermal
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NITROBENZENE TOXICOLOGICAL OVERVIEW

absorption. Ingestion of nitrobenzene may also occur, as the relatively pleasant almond
odour may not discourage individuals from drinking water contaminated with it [1, 3].
Nitrobenzene may be present in the environment close to industrial locations where it is
manufactured or used. It has also been reported as being detectable near hazardous waste
sites. However, the levels of nitrobenzene present in such locations would be expected to be
many times lower than those observed during occupational exposure [1, 3].

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Health Effects of Acute / Single Exposure

Human Data
General toxicity
Nitrobenzene is toxic via inhalation, ingestion and dermal absorption. The principal adverse
health effect following exposure to nitrobenzene is methaemoglobinaemia [1]. There are very
few accounts of human fatalities following nitrobenzene exposure, due to prompt medical
intervention in many case studies [3].
Inhalation
Acute inhalation of nitrobenzene can result in respiratory irritation, headache, nausea,
dizziness, drowsiness, shortness of breath, fatigue, cyanosis and convulsions, due to
methaemoglobinaemia [1, 3, 4]. The symptoms of methaemoglobinaemia may develop within
1 to 4 hours post-exposure [4]. Serious acute exposure to nitrobenzene may lead to
jaundice, renal failure, unconsciousness and can possibly be fatal [1-3, 5]. However, there
are few reports of acute inhalation exposure to nitrobenzene at levels sufficient to cause
fatality [1]. In an early study based on toxic symptoms in factory workers, it was considered
that exposure to 200 ppm would produce serious adverse effects after a 1 hour exposure, or
60 100 ppm for 6 hours [1].
Ingestion
There are numerous reports of poisoning from nitrobenzene in the first half of the 20th
Century when it was widely available as a substitute for oil of bitter almond or as a dye in
shoe polish. Acute oral ingestion of nitrobenzene results in gastric irritation, including nausea
and vomiting, the onset of which may be delayed for up to 12 hours post exposure [1].
Methaemoglobinaemia will develop 1 to 4 hours post-exposure, indicated by cyanosis, the
symptoms of which may be similar to those following inhalation [4]. Severe exposure to
nitrobenzene by ingestion can also give rise to progressive drowsiness and coma which may
prove fatal due to respiratory failure [1].
Dermal / ocular exposure
Dermal exposure to nitrobenzene either from splashes of liquid or contact from vapours may
cause mild irritation [4]. Nitrobenzene will readily undergo dermal absorption and there have
been numerous reports of nitrobenzene toxicity in humans, particularly infants, following skin
contact [1, 3, 4]. Following dermal absorption of sufficient quantities of nitrobenzene the main
adverse effect of nitrobenzene exposure is methaemoglobinaemia similar to that observed
following inhalation and ingestion [1, 2].
Delayed effects following an acute exposure
The onset of methaemoglobinaemia from all routes of exposure may be delayed for 1 to 4
hours depending upon the severity of exposure [1, 3, 4]. In some cases following severe
ingestion of nitrobenzene, haemolytic anaemia may develop around 5 days post exposure
[1].

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Animal and In-Vitro Data

General toxicity
The acute toxicity of nitrobenzene in experimental animals resembles that seen in humans,
with the development of methaemoglobin being the characteristic adverse effect [1, 3, 5, 6].
Inhalation
There are only limited data on acute exposure to nitrobenzene via inhalation. Exposure of
rats to an atmosphere saturated with nitrobenzene vapour for 3 hours resulted in no deaths
over the 14 day observation period. However, exposure for 7 hours resulted in 25% of the
exposed animals (3/12) dying. No information was available on the actual exposure levels
[1].
Ingestion
The oral LD50 for nitrobenzene administered to rats is 600 mg kg-1 body weight [1, 3]. In a
study in which female rats were orally administered 640 mg kg-1 nitrobenzene the percentage
of methaemoglobin formed 30, 60 and 120 mins post-exposure were 11, 19 and 28%,
respectively [1]. Histological changes in the liver and testes were noted at autopsy in rats 2
or 5 days after single doses of nitrobenzene at 200 mg kg-1 and above [1].
Dermal
The dermal LD50 of nitrobenzene in rats has been reported to be 2100 mg kg-1 body weight
[1]. In this dermal LD50 study, the percentage of methaemoglobin formed following an LD50
dose, was found to be 16%, 25% and 35% at 30, 60 and 120 minutes, respectively. The
dermal LD50 of nitrobenzene in rabbits was found to be 760 mg kg-1 body weight [1].
Skin and Eye Irritation
Nitrobenzene only produces slight and transient skin and eye irritation. The skin irritation
potential of nitrobenzene was investigated in male rabbits using the Draize method and was
found to produce only very slight erythema at 24 hours post exposure [1]. Studies
investigating ocular irritation in male rabbits using the Draize method found nitrobenzene to
produce slight irritation at 1 and 24 hours post-exposure, but had resolved completely at 48
hours [1].

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Health Effects of Chronic / Repeated Exposure

Human Data
Inhalation
Long term occupational exposure to nitrobenzene may be associated with similar symptoms
as those observed following excessive levels of acute exposure. The most common adverse
effect following occupational inhalation of nitrobenzene is the formation of
methaemoglobinaemia [1, 3]. A worker exposed to nitrobenzene (no details available on
exposure levels) for 17 months developed severe methaemoglobinaemia, with headache,
nausea, vertigo, confusion and an increased sensitivity to pain (hyperalgesia) from pin-prick.
The worker also had enlarged and tender spleen and liver and abnormal results from liverfunction tests. The exposure and absorption of nitrobenzene were confirmed by the presence
of p-nitrophenol and p-aminophenol in the urine [1]. Chronic exposure to nitrobenzene may
also give rise to the development of haemolytic anaemia and toxic hepatitis [1, 5].
Dermal / Ocular
Very little information was identified relating to chronic occupational dermal exposure of
nitrobenzene. However, as nitrobenzene will undergo rapid dermal absorption the symptoms
are expected to be similar to those observed following chronic inhalation.
Genotoxicity
No studies were identified regarding genotoxic effects following chronic exposure to
nitrobenzene in humans [1, 3]
Carcinogenicity
No data was located regarding the carcinogenicity of nitrobenzene in humans. The
International Agency for Research on Cancer (IARC) has concluded that there is insufficient
evidence for the carcinogenicity of nitrobenzene in humans. However, based on animal
carcinogenicity data, nitrobenzene has been classified overall as possibly carcinogenic to
humans (group 2B) [5].
Reproductive and developmental toxicity
No studies were located regarding reproductive or developmental effects of nitrobenzene in
humans [1, 3].

Animal and In-Vitro Data

Inhalation
The toxicity of nitrobenzene following chronic inhalation was studied in two strains of rat
(Fisher-344 and Sprague Dawley) and B6C3F1 mice, exposed to nitrobenzene at
concentrations up to 125 ppm for 6 hours day-1, 5 days week-1 for 2 weeks. The SpragueDawley rats exposed to 125 ppm nitrobenzene showed severe adverse clinical signs
including rapid shallow breathing and wheezing with 40% lethality at the fourth day of
exposure. All of the mice exposed at the same concentration showed morbidity which
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necessitated the animals being sacrificed on the fourth day of exposure. However, the
Fisher-344 rats tolerated this dose for 2 weeks and showed no adverse clinical signs,
indicating marked strain differences in susceptibility to nitrobenzene [1]. The study also
reported significant concentration-dependent increases in liver, spleen and kidney weights
[1]. Rats exposed to nitrobenzene by inhalation at just 5 ppm (25.15 mg m-3) for 6 hours day1
, 5 days week-1 for 90 days displayed symptoms of methaemoglobinaemia [3].
Ingestion
A study of Fisher-344 rats orally exposed to nitrobenzene at 125 mg kg-1 body weight day-1
for 28 days, showed decreased movement, pale skin, gait abnormalities and decreases in
body weight or body weight gain. In this study, increases in the weights of the liver, spleen
and kidneys were observed along with reductions in the weight of the thymus and the testis
in the males. An increase in liver weights of males was also observed at a low dose of 5 mg
kg-1 body weight day-1 [1]. In another study in Fisher-344 rats, nitrobenzene was
administered (by gavage) for 13 weeks at doses of up to 150 mg kg-1. Some lethality
occurred at the top dose. Clinical signs seen at 75 and 150 mg kg-1 body weight day-1
included ataxia, head tilt, lethargy, trembling, circling, dyspnoea as well as cyanosis of the
extremities. Marked brain lesions were noted at autopsy [1]. A study in which B6C3F1 mice
were administered nitrobenzene orally at up to 300 mg kg-1 body weight day-1 for 13 weeks
showed signs of toxicity including ataxia, lethargy, ataxia, dyspnoea, convulsions, irritability
and rapid head-bobbing movements, some lethality was noted at the highest dose [1].
Dermal
The effects of repeated skin painting of nitrobenzene have been investigated in a number of
studies. In 14 day studies in B6C3F1 mice and Fisher-344 rats, similar effects were seen in
both species. Dose levels in the range of 200 3200 mg kg-1 body weight day-1 were used.
Dose levels of 1600 mg kg-1 body weight day-1 and above resulted in lethality. Clinical signs
reported included ataxia, prostration and dyspnoea. Significant depression of weight gain
was noted in mice at all dose levels. At autopsy there was histological evidence of damage to
the brain, liver, spleen and testes, with mice being less affected than rats [1].
Genotoxicity
Nitrobenzene has been fairly extensively investigated for its genotoxic potential using both invitro and in-vivo studies. The data suggest that it is not a genotoxic carcinogen and does not
possess any significant mutagenic effects in vivo [1].
Nitrobenzene has been investigated for its ability to produce mutations or DNA lesions in two
well established assays. There are numerous reports of studies to investigate the potential of
nitrobenzene to produce gene mutations in vitro in the Ames test using strains of Salmonella
typhimurium, either in the presence or absence of metabolic activation with liver S9 fraction.
Negative results were consistently obtained [1].
Nitrobenzene was also found to be negative for unscheduled DNA synthesis (UDS) when
tested using primary hepatocytes cultured in vitro from either humans or rats [1].
The ability of nitrobenzene to induce chromosome damage has been investigated in vivo.
Negative results were obtained in a micronucleus study using mice and the intraperitoneal
route of exposure. Dose levels of up to 250 mg kg-1 body weight were used, with bone
marrow harvested after 24 and 48 hours and examined for micronucleated polychromatic
erythrocytes. Nitrobenzene has also been investigated in the in-vivo UDS test with rat liver
cells. The compound was given orally at dose levels up to 500 mg kg-1. The study was limited
by the use of a single harvest time, but there was no increase in UDS [1].
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Carcinogenicity
The carcinogenicity of nitrobenzene has been investigated in chronic bioassays using the
inhalation route in rat and mice studies. In one study in rats an increase in the incidence of
hepatocellular neoplasms, thyroid follicular-cell adenomas and adenocarcinomas and renal
tubular-cell adenomas was seen in the treated males. In the treated females there was an
increase in hepatocellular neoplasms and endometrial stromal polyps. In a second study in
which only male rats were used, an increase in the incidence of hepatocellular neoplasm was
seen [5]. In male mice exposed to nitrobenzene an increase in alveolar-bronchial neoplasms
and thyroid follicular-cell adenomas was observed [5].
Overall, the IARC has concluded that there is sufficient evidence for the carcinogenicity of
nitrobenzene in experimental animals [5].
Reproductive and developmental toxicity
Numerous studies have established that nitrobenzene is a testicular toxicant. The most
sensitive end points were sperm count and motility, followed by progressive motility, viability,
presence of abnormal sperm and finally fertility index [1]. In repeated dose studies (13 week)
testicular atrophy was seen in mice at daily doses of 19 mg kg-1 body weight and above, and
in rats at 150 mg kg-1 body weight and above. In a 2 generation reproductive toxicity study in
rats using the inhalation route, 40 ppm (6 hours day-1, 5 days week-1) produced a marked
decrease in fertility index, but was not seen at 20 ppm [1]. These studies suggest that
nitrobenzene may cause reproductive toxicity in males due to testicular atrophy [1, 3, 5].
Studies investigating the developmental toxicity of nitrobenzene in rats and rabbits using the
inhalation route have not observed fetotoxic, embryotoxic or teratogenic effects at
concentrations associated with maternal toxicity [1].

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References
[1]

International Programme on Chemical Safety (IPCS) (2003). Nitrobenzene.

Environmental Health Criteria 230. WHO. Geneva.

[2]

International Programme on Chemical Safety (IPCS) (1993). Nitrobenzene.

International Chemical Safety Card: 0065. WHO. Geneva.

[3]

Agency for Toxic Substances and Disease Registry (ATSDR) (1990). Toxicological
Profile for Nitrobenzene. US department of Health and Human Services. Atlanta, US.

[4]

National Poisons Information Service (NPIS) (2007). Nitrobenzene. TOXBASE.

[5]

International Agency for the Research on Cancer (IARC) (1996). Nitrobenzene. Vol
65. IARC. Lyon.

[6]

Weisburger, E. K. and Hudson, V. W. (2001). Aromatic nitro and amino compounds,

Chapter fifty-seven. John Wiley & Sons, Inc. New York, NY.

This document from the HPA Centre for Radiation, Chemical and Environmental Hazards reflects
understanding and evaluation of the current scientific evidence as presented and referenced in this
document.

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