<b>Bijsluiter</b>. De hyperlink naar het originele document werkt niet meer. Daarom laat Woogle de tekst zien die in dat document stond. Deze tekst kan vreemde foutieve woorden of zinnen bevatten en de opmaak kan verdwenen of veranderd zijn. Dit komt door het zwartlakken van vertrouwelijke informatie of doordat de tekst niet digitaal beschikbaar was en dus ingescand en vervolgens via OCR weer ingelezen is. Voor het originele document, neem contact op met de Woo-contactpersoon van het bestuursorgaan.<br><br>====================================================================== Pagina 1 ======================================================================

<pre>2-Nitroanisole
Evaluation of the carcinogenicity and genotoxicity
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<pre>Gezondheidsraad                                          Vo o r z i t t e r
Health Council of the Netherlands
Aan de minister van Sociale Zaken en Werkgelegenheid
Onderwerp               : Aanbieding advies 2-Nitroanisole
Uw kenmerk              : DGV/MBO/U-932542
Ons kenmerk             : U-5135/JR/pg/246-H12
Bijlagen                :1
Datum                   : 1 april 2008
Geachte minister,
Graag bied ik u hierbij het advies aan over de kankerverwekkendheid van 2-nitroanisole.
Het maakt deel uit van een uitgebreide reeks waarin kankerverwekkende stoffen worden
geclassificeerd volgens richtlijnen van de Europese Unie. Het gaat om stoffen waaraan
mensen tijdens de beroepsmatige uitoefening kunnen worden blootgesteld.
     Het advies is opgesteld door een vaste subcommissie van de Commissie Gezondheid en
beroepsmatige blootstelling aan stoffen (GBBS), de Subcommissie Classificatie van carci-
nogene stoffen. Het advies is voorgelegd aan de Commissie GBBS en vervolgens getoetst
door de Beraadsgroep Gezondheid en omgeving van de Gezondheidsraad.
Ik heb dit advies vandaag ter kennisname toegezonden aan de minister van Volksgezond-
heid, Welzijn en Sport en de minister van Volkshuisvesting, Ruimtelijke Ordening en
Milieubeheer.
Hoogachtend,
prof. dr. J.A. Knottnerus
Bezoekadres                                                                 Postadres
Parnassusplein 5                                                            Postbus 16052
2 5 11 V X D e n          Haag                                              2500 BB Den            Haag
Te l e f o o n ( 0 7 0 ) 3 4 0 6 6 3 1                                      Te l e f a x ( 0 7 0 ) 3 4 0 7 5 2 3
E - m a i l : j o l a n d a . r i j n k e l s @ g r. n l                    w w w. g r. n l
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<pre>2-Nitroanisole
Evaluation of the carcinogenicity and genotoxicity
Subcommittee on the classification of carcinogenic substances of the
Dutch Expert Committee on Occupational Standards,
a committee of the Health Council of the Netherlands
to:
the Minister of Social Affairs and Employment
No. 2008/02OSH, The Hague, April 1, 2008
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<pre>The Health Council of the Netherlands, established in 1902, is an independent
scientific advisory body. Its remit is “to advise the government and Parliament on
the current level of knowledge with respect to public health issues...” (Section
22, Health Act).
     The Health Council receives most requests for advice from the Ministers of
Health, Welfare & Sport, Housing, Spatial Planning & the Environment, Social
Affairs & Employment, and Agriculture, Nature & Food Quality. The Council
can publish advisory reports on its own initiative. It usually does this in order to
ask attention for developments or trends that are thought to be relevant to gov-
ernment policy.
     Most Health Council reports are prepared by multidisciplinary committees of
Dutch or, sometimes, foreign experts, appointed in a personal capacity. The
reports are available to the public.
                 The Health Council of the Netherlands is a member of the European
                 Science Advisory Network for Health (EuSANH), a network of science
                 advisory bodies in Europe.
                 The Health Council of the Netherlands is a member of the International Network
                 of Agencies for Health Technology Assessment (INAHTA), an international
                 collaboration of organisations engaged with health technology assessment.
 I NA HTA
This report can be downloaded from www.healthcouncil.nl.
Preferred citation:
Health Council of the Netherlands. 2-Nitroanisole; Evaluation of the carcinoge-
nicity and genotoxicity. The Hague: Health Council of the Netherlands, 2008;
publication no. 2008/02OSH.
all rights reserved
ISBN: 978-90-5549-687-7
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<pre>    Contents
    Samenvatting 9
    Executive summary 11
1   Scope 13
1.1 Background 13
1.2 Committee and procedures 13
1.3 Data 14
2   General information 15
2.1 Identity and physico-chemical properties 15
2.2 IARC classification 16
3   Carcinogenicity 17
3.1 Observations in humans 17
3.2 Carcinogenicity studies in animals 17
4   Mutagenicity and genotoxicity 21
4.1 In vitro assays 21
4.2 In vivo assays 21
    Contents                                    7
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<pre>5   Classification 23
5.1 Evaluation of data on carcinogenicity and genotoxicity 23
5.2 Recommendation for classification 24
    References 25
    Request for advice 29
    Annexes 29
A   The committee 31
B   Comments on the public review draft 33
C   IARC Monograph 35
D   Carcinogenic classification of substances by the committee 37
E   Guideline 93/21/EEG of the European Union 39
8   2-Nitroanisole
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<pre>Samenvatting
Op verzoek van de minister van Sociale Zaken en Werkgelegenheid evalueert en
beoordeelt de Gezondheidsraad de kankerverwekkende eigenschappen van stof-
fen waaraan mensen tijdens het uitoefenen van hun beroep kunnen worden bloot-
gesteld. De evaluatie en beoordeling worden verricht door de subcommissie
Classificatie van Carcinogene Stoffen van de Commissie Gezondheid en
Beroepsmatige Blootstelling aan Stoffen van de Raad, hierna kortweg aangeduid
als de commissie. In het voorliggende advies neemt de commissie 2-nitroanisol
onder de loep. 2-Nitroanisol wordt gebruikt bij de synthese van kleurstoffen en
als intermediair bij productie van geneesmiddelen.
Op basis van de beschikbare gegevens leidt de commissie af dat 2-nitroanisol
beschouwd moet worden als kankerverwekkend voor de mens. Dit is vergelijk-
baar met een classificatie in categorie 2 volgens de richtlijnen van de Europese
Unie. De commissie is verder van mening dat 2-nitroanisol een stochastisch
genotoxisch werkingsmechanisme heeft.
Samenvatting                                                                     9
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<pre>10 2-Nitroanisole</pre>

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<pre>Executive summary
At request of the Minister of Social Affairs and Employment, the Health Council
of the Netherlands evaluates and judges the carcinogenic properties of sub-
stances to which workers are occupationally exposed. The evaluation is per-
formed by the subcommittee on Classifying Carcinogenic Substances of the
Dutch Expert Committee on Occupational Standards of the Health Council, here-
after called the committee. In this report, the committee evaluated 2-nitroanisole.
2-Nitroanisole is used in the synthesis of azo dyes, and as an intermediate for the
preparation of pharmaceuticals.
Based on the available information, the committee is of the opinion that 2-
nitroanisole should be considered as carcinogenic to humans. This recommenda-
tion is comparable to the EU classification in category 2. The committee is fur-
thermore of the opinion that 2-nitroanisole acts by a stochastic genotoxic
mechanism.
Executive summary                                                                   11
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<pre>12 2-Nitroanisole</pre>

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<pre>Chapter 1
        Scope
1.1     Background
        In the Netherlands a special policy is in force with respect to occupational use
        and exposure to carcinogenic substances. Regarding this policy, the Minister of
        Social Affairs and Employment has asked the Health Council of the Netherlands
        to evaluate the carcinogenic properties of substances, and to propose a classifica-
        tion (see Annex A). The assessment and the proposal for a classification are
        expressed in the form of standard sentences (see Annex E). The criteria used for
        classification are partly based on an EU-directive (see Annex F). In addition to
        classifying substances, the Health Council also assesses the genotoxic properties
        of the substance in question.
        This report contains the evaluation of the carcinogenicity of 2-nitroanisole.
1.2     Committee and procedures
        The evaluation is performed by the subcommittee on Classifying Carcinogenic
        Substances of the Dutch Expert Committee on Occupational Standards of the
        Health Council, hereafter called the committee. The members of the committee
        are listed in Annex B. The first draft was prepared by I.A. van de Gevel and M.I.
        Willems, from the Department of Occupational Toxicology of the TNO Nutrition
        Scope                                                                               13
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<pre>    and Food Research, by contract with the Ministry of Social Affairs and Employ-
    ment.
         In 2007 the President of the Health Council released a draft of the report for
    public review. The individuals and organisations that commented on the draft are
    listed in Annex C. The committee has taken these comments into account in
    deciding on the final version of the report.
1.3 Data
    The evaluation and recommendation of the committee is standardly based on sci-
    entific data, which are publicly available. The starting points of the committees’
    reports are, if possible, the monographs of the International Agency for Research
    on Cancer (IARC). This means that the original sources of the studies, which are
    mentioned in the IARC-monograph, are reviewed only by the committee when
    these are considered most relevant in assessing the carcinogenicity and genotox-
    icity of the substance in question. In the case of 2-nitroanisole, such an IARC-
    monograph is available, of which the summary and conclusion of IARC is
    inserted in Annex D.
         More recently published data were retrieved from the online databases Med-
    line, Toxline, Chemical Abstracts, and RTECS. The last updated online search
    was in June 2007. The new relevant data were included in this report.
14  2-Nitroanisole
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<pre>Chapter 2
        General information
2.1     Identity and physico-chemical properties
        2-Nitroanisole is primarily used as a precursor of o(rtho)-anisidine by nitro-
        reduction.1,2 As such, it is used for the synthesis of azo dyes, and as an intermedi-
        ate for the preparation of pharmaceuticals.1,2 Occupational exposure may occur
        during manufacturing of azo dyes and pharmaceuticals.
            Below is given the identity and some of its physical and chemical properties.1
        Chemical name              :  2-nitroanisole
        CAS registry number        :  91-23-6
        EEC number                 :  609-047-00-7
        EINECS number              :  202-052-1
        IUPAC name                 :  ortho-Nitroanisole
        Synonyms                   :  2-Methoxynitrobenzene; 2-methoxy-1-nitrobenzene; ortho-
                                      nitroanisole; ortho-nitrobenzene methyl ether; 2-nitromethoxy-
                                      benzene; ortho-nitromethoxybenzene; 1-nitro-2-methoxybenzene;
                                      ortho-nitrophenyl methyl ether
        Description                :  colourless to yellowish liquid
        Application                :  2-Nitroanisole is reduced (using a H2-catalyst or iron-formic acid)
                                      to ortho-anisidine or to ortho-dianisidine, both of which are
                                      important as dye intermediates. 2-Nitroanisole has also been used
                                      as an intermediate for various pharmaceuticals (IARC96)
        Molecular formula          :  C7H7NO3
        General information                                                                               15
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<pre>    Structure                  :
    Molecular weight           : 153.13
    Boiling point              : 277 °C
    Melting point              : 10.5 °C
    Relative density (air = 1) : 1.254 at 20 °C/4 °C
    Vapour pressure            : 4 Pa at 30 °C
    Solubility                 : moderately soluble in warm water (1.69 g/L at 30 oC); soluble in
                                 ethanol and diethyl ether
    Stability                  : Explosive reaction with sodium hydroxide and zinc
    Log Pow                    : 1.73
    Conversion factors (20°C)  : 1 ppm = 6.35 mg/m3 air
                                 1 mg/m3 = 0.15 ppm
    Risk and safety phrases    : R22: harmful if swallowed
                                 R45: may cause cancer
                                 S45: in case of accident or if you feel unwell, seek medical advice
                                 immediately (show label where possible)
                                 S53: avoid exposure – obtain special instructions before use
    EC classification          : Carcinogenic category 2
2.2 IARC classification
    In 1996, IARC concluded that there is inadequate evidence in humans for the
    carcinogenicity of 2-nitroanisole, but that there is sufficient evidence in experi-
    mental animals.1 Therefore, it classified the agent in group 2B, indicating that it
    is possibly carcinogenic to humans.
16  2-Nitroanisole
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<pre>Chapter 3
        Carcinogenicity
3.1     Observations in humans
        No data were available to evaluate the carcinogenicity of 2-nitroanisole in
        humans.
3.2     Carcinogenicity studies in animals
        Groups of 60 male and 60 female F344 rats were fed 2-nitroanisole-enriched
        diets for 2 years.2,3 The diets contained concentrations of 2-nitroanisole of 0, 222,
        666, or 2,000 ppm (mg/kg diet). Feed and water were freely available. Ten rats
        per group were killed at week 65 for interim evaluation. In males receiving 2,000
        ppm in the diet, the survival rate and the mean body weights were lower than
        controls; for all other groups these were comparable with controls. No increases
        in incidences of neoplasms were observed in exposed animals. However, 2-
        nitroanisole was associated with nonneoplastic lesions of the kidneys and fore-
        stomach.
            In a second experiment by the same research institute, groups of 60 male and
        60 female F344 rats were given 2-nitroanisole-enriched diets for only 27 weeks,
        after which they received control diets for up to an additional 76 weeks (total
        study duration, 2 years).2,3 The diets contained concentrations of 2-nitroanisole of
        0, 6,000, or 18,000 ppm (mg/kg diet). During the experiment four interim evalu-
        ations were performed, leaving 20 animals per group per sex alive for the total
        Carcinogenicity                                                                       17
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<pre>   duration of the experiment. Survival rates, mean body weight and food consump-
   tion of exposed animals were significantly reduced compared to controls. Fur-
   thermore, increased incidences of neoplasms in the urinary bladder, kidneys,
   forestomach, and large intestines were found in animals fed 6,000 and 18,000
   ppm 2-nitroanisole. Details on the type of lesions are shown in Table 3.1 on the
   next page.
       The same research group used also male and female B6C3F1 mice under the
   same experimental conditions as in the first two-year rat study.2,3 The mice were
   given 2-nitroanisole in the diet at doses of 0, 666, 2,000, or 6,000 ppm. No treat-
   ment related changes in survival were observed. However, the body weights of
   animals receiving 2,000 and 6,000 ppm were significantly reduced compared to
   controls. Significantly increased incidences of nonneoplastic lesions of the liver
   were found in all exposed groups compared to controls. The only neoplastic
   lesions were found in the liver of male mice. It concerned hepatocellular ade-
   nomas and carcinomas in males that received 2,000 or 6,000 ppm in their diet
   (control, 21/50; 666 ppm, 33/50; 2,000 ppm, 46/50; 6,000 ppm, 34/50; ade-
   nomas, carcinomas and hepatoblastomas taken together). In female mice, no neo-
   plastic lesions were observed.
       No animal data were available on inhalation exposure or other routes of
   exposure.
   Table 3.1 Neoplastic lesions in F344 rats, which were fed 2-nitroanisole enriched diets for 27
   weeks. Animals were examined microscopically after an additional 76 weeks.2,3
   Dose administered                           0 ppm             6,000 ppm         12,000 ppm
   Males
   Urinary bladder
     No. of animals examined                   21               27                 34
     Transitional epithelium carcinoma          0                23**              33**
     Transitional epithelium papilloma          0                 3                 1
     Squamous cell carcinoma                    0                 0                 5
     Squamous cell papilloma                    0                 0                 4
     Sarcomaa                                   0                 1                 7**
   The kidneys
     No. of animals examined                   21               27                 34
     Transitional cell papilloma                0                 0                 3
     Transitional cell carcinoma                0                 1                 6*
   Large intestine
     No. of animals examined                   21               27                 34
     Adenomatous polyp                          0                21**              24**
     Carcinoma                                  0                 0                 4
18 2-Nitroanisole
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<pre>Table 3.1 Continued.
Dose administered                         0 ppm           6,000 ppm        12,000 ppm
Females
Urinary bladder
  No. of animals examined                 20              20               34
  Transitional epithelium carcinoma         0             18**             32**
  Transitional epithelium papilloma         0              1                 1
  Squamous cell carcinoma                   0              0                 1
  Squamous cell papilloma                   0              0                 4
  Sarcoma                                   0              2               12**
The kidneys
  No. of animals examined                 22              20               34
  Transitional cell papilloma               0              0                 1
  Transitional cell carcinoma               0              0                 1
Large intestine
  No. of animals examined                 22              20               34
  Adenomatous polyp                         0              5*              17**
  Carcinoma                                 0              0                 2
a   Includes a leiomyosarcoma in one 6,000 ppm and two 18,000 females, and a fibrosarcoma in
    one 18,000 ppm female.
*   p< 0.05 versus controls; ** p< 0.01 versus controls.
Carcinogenicity                                                                              19
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<pre>20 2-Nitroanisole</pre>

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<pre>Chapter 4
        Mutagenicity and genotoxicity
4.1     In vitro assays
        2-Nitroanisole was tested in the Salmonella mutagenicity bioassay.1,4,5 Variable
        responses were found depending on the type of strain used and doses applied.
        Overall, positive outcomes were observed using TA100 (with and without an
        exogenous metabolic system), and in some but not all tests using TA1535,
        TA1538, and TA98. Furthermore, 2-nitroanisole induced mutations in Bacillus
        subtilis rec H97 and M45 strains, in the absence of a metabolic system.1,4 Muta-
        tions at the TK locus were also found of mouse lymphoma L1578Y cells.1,4
             Using human hepatic cytosolic samples, the 32P-post-labeling technique, and
        3
          H-labeled 2-nitroanisole, Stiborová et al. (2004) showed that 2-nitroanisole was
        activated to form DNA-adducts in vitro.6
             Concerning the clastogenic potential, 2-nitroanisole increased the frequency
        of chromosomal aberrations and sister chromatid exchanges in Chinese hamster
        cells.1,4,7
4.2     In vivo assays
        Hengstler et al. (1995) found increased levels of DNA single-strand breaks in
        mononuclear blood cells of 16 fire fighters, who were exposed to a mixture of 2-
        nitroanisole and other chemical agents.8 Three months after the accident, levels
        of single-strand breaks were decreased to reference levels. Since during the fire
        Mutagenicity and genotoxicity                                                      21
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<pre>   accident also other chemicals were released in the environment, it is difficult to
   assess whether 2-nitroanisole was the only agent that was responsible for the
   observed effects.
        Stiborová et al. (2004) also investigated the capacity of 2-nitroanisole to
   form DNA adducts in vivo.6 Six male Wistar rats were given intraperitoneal
   injection of 2-nitroanisole (0.15 mg/kg bw), once a day for five consecutive days.
   An additional two animals served as vehicle-controls. Animals were killed 24
   hours after the last injection, and various organs were removed and prepared for
   further analysis. Using the 32P-post-labeling technique, in exposed animals, the
   authors found the highest levels of treatment-related DNA-adducts in the urinary
   bladder, followed by the liver, kidney and spleen. The types of adducts found in
   animals were similar to those formed in vitro. No adducts were found in the
   lungs, brain and heart, nor were there adducts found in the organs of control ani-
   mals.
        Esmaeili et al. (2006) used a host-mediated in vitro/in vivo system to study 2-
   nitroanisole-induced transformations of peritoneal macrophages.9 Groups of five
   male NMRI mice were given a single intraperitoneal injection of 0, 1.3, 65 or
   130 mg/kg bw (0%, 0.1%, 5%, or 10% of LD50, respectively). The transforming
   potential of the agent increased dose-dependently. Moreover, the investigators
   could establish an immortal cell line from the peritoneal macrophages, obtained
   from mice at the highest dose group. The immortal cell line was further tested for
   its tumour-inducing potency by subcutaneous injection of these cells on nude
   mice (nu/nu; number of animals not specified). This resulted in visible tumour
   formation, three weeks after the injection. Microscopic examination of the
   tumour tissue indicated typical signs of malignancy of mesenchymal origin.
   The cell line was also tested on activity of the proto-oncogenes c-jun, c-myc, and
   c-fos. Dysfunction of these proto-oncogenes has been shown in diverse human
   tumours, like lymphomas and sarcomas. The transformed cells showed a signifi-
   cant down-regulation of c-myc and c-fos, and an overexpression of c-jun.
22 2-Nitroanisole
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<pre>Chapter 5
        Classification
5.1     Evaluation of data on carcinogenicity and genotoxicity
        No data on the genotoxicity and carcinogenicity of 2-nitroanisole in humans
        were available, nor were there any data available on inhalation exposure in ani-
        mals.
             Oral administration of 2-nitroanisole for up to two years increased the inci-
        dence of tumours in the urinary bladder (rats), large intestines (rats), the kidneys
        (rats), and the liver (mice) compared to nontreated animals. These findings give
        sufficient evidence that oral exposure to 2-nitroanisole results in tumour devel-
        opment.
             The number of studies on the mutagenic and genotoxic potential of the agent
        is limited. Overall, 2-nitroanisole induced mutations in bacteria and in mamma-
        lian cells, and showed to be clastogenic in vitro. It was also able to transform nor-
        mal peritoneal macrophages into immortal cells. These immortal cells had
        altered expression of proto-oncogenes and, furthermore, formed tumours at the
        injection site. Based on these mutagenicity and genotoxicity data, the committee
        considers 2-nitroanisole as a genotoxic carcinogen that acts by a stochastic
        mechanism.
             The committee did not find indications that the observations in animals, and
        the proposed carcinogenic mechanism would not occur in humans.
        Classification                                                                        23
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<pre>5.2 Recommendation for classification
    The committee is of the opinion that 2-nitroanisole should be considered as car-
    cinogenic to humans. This recommendation is comparable to the EU classifica-
    tion in category 2. The committee is furthermore of the opinion that 2-
    nitroanisole acts by a stochastic genotoxic mechanism.
24  2-Nitroanisole
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<pre>  References
1 International Agency for Research on Cancer. 2-Nitroanisole. IARC Monogr Eval. Carcinog. Risks
  Hum., Volume 65: pp 369-380; 1996: 65.
2 National Toxicity Program. NTP Toxicology and Carcinogenesis Studies of o-Nitroanisole (CAS No.
  91-23-6) in F344 Rats and B6C3F1 Mice (Feed Studies). Natl.Toxicol.Program.Technical Report
  Series, Volume 416; 1993: 416.
3 Irwin RD, Chhabra R, Eustis S, Pinter A, Prejean JD. Tumors of the bladder, kidney, and intestine of
  F344 rats and liver of B6C3F1 mice administered o-nitroanisole in feed. Fundam Appl Toxicol 1996;
  30(1): 1-12.
4 Deutsche Fortschungsgemeinschaft. Gesundheitsschädliche Arbeitsstoffe; Toxikologisch-
  arbeitsmedizinische begründung von MAK-Werten. VCH; 1994.
5 European Chemicals Bureau (ECB). IUCLID dataset, CD-ROM edition 2000: 2-Nitroanisole (202-
  052-1). Brussels: European Commission; 2000.
6 Stiborova M, Miksanova M, Smrcek S, Bieler CA, Breuer A, Klokow KA et al. Identification of a
  genotoxic mechanism for 2-nitroanisole carcinogenicity and of its carcinogenic potential for humans.
  Carcinogenesis 2004; 25(5): 833-840.
7 Galloway SM, Armstrong MJ, Reuben C, Colman S, Brown B, Cannon C et al. Chromosome
  aberrations and sister chromatid exchanges in Chinese hamster ovary cells: evaluations of 108
  chemicals. Environ Mol Mutagen 1987; 10 Suppl 10: 1-175.
8 Hengstler JG, Fuchs J, Bolm-Audorff U, Meyer S, Oesch F. Single-strand breaks in deoxyribonucleic
  acid in fire fighters accidentally exposed to o-nitroanisole and other chemicals. Scand J Work
  Environ Health 1995; 21(1): 36-42.
  References                                                                                           25
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<pre>9  Esmaeili A, Schlatterer K, Demirhan I, Schlatterer B, Nauck M, Chandra P et al. Tumorigenic
   potential and the molecular mechanism of the carcinogenic effect exerted by 2-nitroanisole.
   Anticancer Res 2006; 26(6B): 4203-4212.
26 2-Nitroanisole
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<pre>A Request for advice
B The committee
C Comments on the public review draft
D IARC Monograph
E Carcinogenic classification of substances by the committee
F Guideline 93/21/EEG of the European Union
  Annexes
                                                             27
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<pre>28 2-Nitroanisole</pre>

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<pre>Annex A
      Request for advice
      In a letter dated October 11, 1993, ref DGA/G/TOS/93/07732A, to, the State
      Secretary of Welfare, Health and Cultural Affairs, the Minister of Social Affairs
      and Employment wrote:
      Some time ago a policy proposal has been formulated, as part of the simplification of the governmen-
      tal advisory structure, to improve the integration of the development of recommendations for health
      based occupation standards and the development of comparable standards for the general population.
      A consequence of this policy proposal is the initiative to transfer the activities of the Dutch Expert
      Committee on Occupational Standards (DECOS) to the Health Council. DECOS has been established
      by ministerial decree of 2 June 1976. Its primary task is to recommend health based occupational
      exposure limits as the first step in the process of establishing Maximal Accepted Concentrations
      (MAC-values) for substances at the work place.
      In an addendum, the Minister detailed his request to the Health Council as fol-
      lows:
      The Health Council should advice the Minister of Social Affairs and Employment on the hygienic
      aspects of his policy to protect workers against exposure to chemicals. Primarily, the Council should
      report on health based recommended exposure limits as a basis for (regulatory) exposure limits for air
      quality at the work place. This implies:
      •    A scientific evaluation of all relevant data on the health effects of exposure to substances using a
           criteria-document that will be made available to the Health Council as part of a specific request
      Request for advice                                                                                        29
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<pre>       for advice. If possible this evaluation should lead to a health based recommended exposure limit,
       or, in the case of genotoxic carcinogens, a ‘exposure versus tumour incidence range’ and a calcu-
       lated concentration in air corresponding with reference tumour incidences of 10-4 and 10-6 per
       year.
   •   The evaluation of documents review the basis of occupational exposure limits that have been
       recently established in other countries.
   •   Recommending classifications for substances as part of the occupational hygiene policy of the
       government. In any case this regards the list of carcinogenic substances, for which the classifica-
       tion criteria of the Directive of the European Communities of 27 June 1967 (67/548/EEG) are
       used.
   •   Reporting on other subjects that will be specified at a later date.
   In his letter of 14 December 1993, ref U 6102/WP/MK/459, to the Minister of
   Social Affairs and Employment the President of the Health Council agreed to
   establish DECOS as a Committee of the Health Council.
30 2-Nitroanisole
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<pre>Annex B
      The committee
      •   G..J. Mulder, chairman
          emeritus professor of toxicology, Leiden University, Leiden
      •   P.J. Boogaard
          toxicologist, SHELL International BV, The Hague
      •   Ms. M.J.M. Nivard
          molecular biologist and genetic toxicologist, Leiden University Medical Cen-
          ter, Leiden
      •   G.M.H. Swaen
          epidemiologist, Dow Chemicals NV, Terneuzen
      •   R.A. Woutersen
          toxicologic pathologist, TNO Quality of Life, Zeist
      •   A.A. van Zeeland
          professor of molecular radiation dosimetry and radiation mutagenesis, Uni-
          versity Medical Center, Leiden
      •   E.J.J. van Zoelen
          professor of cell biology, Radboud University Nijmegen, Nijmegen
      •   J.M. Rijnkels, scientific secretary
          Health Council of the Netherlands, The Hague
      The committee consulted an additional expert, Prof. dr. G. Mohn, working at
      Department of Radiation Genetics and Chemical Mutagenesis of the University
      of Leiden, with respect to the genotoxic data.
      The committee                                                                    31
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<pre>   The Health Council and interests
   Members of Health Council Committees are appointed in a personal capacity
   because of their special expertise in the matters to be addressed. Nonetheless, it
   is precisely because of this expertise that they may also have interests. This in
   itself does not necessarily present an obstacle for membership of a Health Coun-
   cil Committee. Transparency regarding possible conflicts of interest is nonethe-
   less important, both for the President and members of a Committee and for the
   President of the Health Council. On being invited to join a Committee, members
   are asked to submit a form detailing the functions they hold and any other mate-
   rial and immaterial interests which could be relevant for the Committee’s work.
   It is the responsibility of the President of the Health Council to assess whether
   the interests indicated constitute grounds for non-appointment. An advisorship
   will then sometimes make it possible to exploit the expertise of the specialist
   involved. During the establishment meeting the declarations issued are dis-
   cussed, so that all members of the Committee are aware of each other’s possible
   interests.
32 2-Nitroanisole
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<pre>Annex C
      Comments on the public review draft
      A draft of the present report was released in 2007 for public review. The follow-
      ing organisations and persons have commented on the draft document:
      • E. González-Fernández, Ministerio de Trabajo y Asuntos Sociales, Spain;
      • R.D. Zumwalde, National Institute for Occupational Safety and Health, the
          USA.
      Comments on the public review draft                                               33
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<pre>34 2-Nitroanisole</pre>

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<pre>Annex D
      IARC Monograph
      Vol.: 65 (1996) (p. 369)1
      CAS No.: 91-23-6
      Chem. Abstr. Name: 1-Methoxy-2-nitrobenzene
      Summary of Data Reported and Evaluation
      Exposure data
      2-Nitroanisole is produced by the reaction of methanolic sodium hydroxide with
      2-chloronitrobenzene. It is mainly used in the production of the dye intermedi-
      ates ortho-anisidine and ortho-dianisidine. Human exposure may occur during its
      production and use.
      Human carcinogenicity data
      No data on the carcinogenicity of 2-nitroanisole in humans were available to the
      Working Group.
      Animal carcinogenicity studies
      2-Nitroanisole was tested for carcinogenicity by oral administration in one study
      in mice and in two studies in rats. In mice, the incidence of hepatocellular ade-
      IARC Monograph                                                                    35
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<pre>   nomas was increased in males and females, and that of hepatoblastomas was
   increased in males. In one study in rats, the incidence of mononuclear-cell leu-
   kaemia was increased in males and females. In the second study, which used a
   shorter duration of treatment but higher doses, increases were seen in the inci-
   dences of tumours of the urinary bladder, the large intestine and the kidney.
   Other relevant data
   No human data were available on the metabolism of 2-nitroanisole.
       In rats, 2-nitroanisole is absorbed after oral administration, and the major
   route of its rapid elimination is the urine. The predominant metabolic pathway
   involves the formation of 2-nitrophenol, with its subsequent conjugation with
   sulfate and glucuronic acid. 2-Nitroanisole causes methaemoglobinaemia follow-
   ing dietary administration of high doses to rats and mice. Pathological lesions
   observed in rats occurred in the urinary bladder, spleen, kidney and liver. In
   mice, 2-nitroanisole causes hypertrophy in the liver.
       2-Nitroanisole is mutagenic in bacteria. In single studies, it induced muta-
   tions, sister chromatid exchange and a low frequency of chromosomal aberra-
   tions in cultured mammalian cells.
   Evaluation
   There is inadequate evidence in humans for the carcinogenicity of 2-nitroanisole.
       There is sufficient evidence in experimental animals for the carcinogenicity
   of 2-nitroanisole.
   Overall evaluation
   2-Nitroanisole is possibly carcinogenic to humans (Group 2B).
36 2-Nitroanisole
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<pre>Annex         E
              Carcinogenic classification of
              substances by the committee
The committee expresses its conclusions in the form of standard phrases:
Judgment of the committee                                                                       Comparable with EU class
This compound is known to be carcinogenic to humans                                             1
•   It is stochastic or non-stochastic genotoxic
•   It is non-genotoxic
•   Its potential genotoxicity has been insufficiently investigated. Therefore, it is unclear
    whether it is genotoxic
This compound should be regarded as carcinogenic to humans                                      2
•   It is stochastic or non-stochastic genotoxic
•   It is non-genotoxic
•   Its potential genotoxicity has been insufficiently investigated. Therefore, it is unclear
    whether it is genotoxic
This compound is a suspected human carcinogen.                                                  3
•   This compound has been extensively investigated. Although there is insufficient evidence    (A)
    for a carcinogenic effect to warrant a classification as ‘known to be carcinogenic to
    humans’ or as ‘should be regarded as carcinogenic to humans’, they indicate that there is
    cause for concern.
•   This compound has been insufficiently investigated. While the available data do not war-    (B)
    rant a classification as ‘known to be carcinogenic to humans’ or as ‘should be regarded as
    carcinogenic to humans’, they indicate that there is a cause for concern.
This compound cannot be classified                                                              not classifiable
•   There is a lack of carcinogenicity and genotoxicity data.
•   Its carcinogenicity is extensively investigated. The data indicate sufficient evidence sug-
    gesting lack of carcinogenicity.
              Carcinogenic classification of substances by the committee                                             37
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<pre>Annex F
      Guideline 93/21/EEG of the European
      Union
      4.2            Criteria for classification, indication of danger, choice of risk phrases
      4.2.1          Carcinogenic substances
      For the purpose of classification and labelling, and having regard to the current state of knowledge,
      such substances are divided into three categories:
      Category 1:
      Substances known to be carcinogenic to man.
      There is sufficient evidence to establish a causal association between human exposure to a substance
      and the development of cancer.
      Category 2:
      Substances which should be regarded as if they are carcinogenic to man.
      There is sufficient evidence to provide a strong presumption that human exposure to a substance may
      result in the development of cancer, generally on the basis of:
      •    appropriate long-term animal studies
      •    other relevant information.
      Guideline 93/21/EEG of the European Union                                                             39
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<pre>   Category 3:
   Substances which cause concern for man owing to possible carcinogenic effects but in
   respect of which the available information is not adequate for making a satisfactory assess-
   ment.
   There is some evidence from appropriate animal studies, but this is insufficient to place the substance
   in Category 2.
   4.2.1.1       The following symbols and specific risk phrases apply:
   Category 1 and 2:
   T; R45 May cause cancer
   However for substances and preparations which present a carcinogenic risk only when inhaled, for
   example, as dust, vapour or fumes, (other routes of exposure e.g. by swallowing or in contact with
   skin do not present any carcinogenic risk), the following symbol and specific risk phrase should be
   used:
   T; R49 May cause cancer by inhalation
   Category 3:
   Xn; R40 Possible risk of irreversible effects
   4.2.1.2       Comments regarding the categorisation of carcinogenic substances
   The placing of a substance into Category 1 is done on the basis of epidemiological data; placing into
   Categories 2 and 3 is based primarily on animal experiments.
   For classification as a Category 2 carcinogen either positive results in two animal species should be
   available or clear positive evidence in one species; together with supporting evidence such as geno-
   toxicity data, metabolic or biochemical studies, induction of benign tumours, structural relationship
   with other known carcinogens, or data from epidemiological studies suggesting an association.
40 2-Nitroanisole
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<pre>Category 3 actually comprises 2 sub-categories:
a    substances which are well investigated but for which the evidence of a tumour-inducing effect is
     insufficient for classification in Category 2. Additional experiments would not be expected to
     yield further relevant information with respect to classification.
b    substances which are insufficiently investigated. The available data are inadequate, but they
     raise concern for man. This classification is provisional; further experiments are necessary
     before a final decision can be made.
For a distinction between Categories 2 and 3 the arguments listed below are relevant which reduce
the significance of experimental tumour induction in view of possible human exposure. These argu-
ments, especially in combination, would lead in most cases to classification in Category 3, even
though tumours have been induced in animals:
•    carcinogenic effects only at very high levels exceeding the 'maximal tolerated dose'. The maxi-
     mal tolerated dose is characterized by toxic effects which, although not yet reducing lifespan, go
     along with physical changes such as about 10% retardation in weight gain;
•    appearance of tumours, especially at high dose levels, only in particular organs of certain species
     is known to be susceptible to a high spontaneous tumour formation;
•    appearance of tumours, only at the site of application, in very sensitive test systems (e.g. i.p. or
     s.c. application of certain locally active compounds);
•    if the particular target is not relevant to man;
•    lack of genotoxicity in short-term tests in vivo and in vitro;
•    existence of a secondary mechanism of action with the implication of a practical threshold above
     a certain dose level (e.g. hormonal effects on target organs or on mechanisms of physiological
     regulation, chronic stimulation of cell proliferation;
•    existence of a species - specific mechanism of tumour formation (e.g. by specific metabolic
     pathways) irrelevant for man.
For a distinction between Category 3 and no classification arguments are relevant which exclude a
concern for man:
•    a substance should not be classified in any of the categories if the mechanism of experimental
     tumour formation is clearly identified, with good evidence that this process cannot be extrapo-
     lated to man;
•    if the only available tumour data are liver tumours in certain sensitive strains of mice, without
     any other supplementary evidence, the substance may not be classified in any of the categories;
•    particular attention should be paid to cases where the only available tumour data are the occur-
     rence of neoplasms at sites and in strains where they are well known to occur spontaneously with
     a high incidence.
Guideline 93/21/EEG of the European Union                                                                 41
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<br><br>