<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>Chloroform
Evaluation of the effects on reproduction, recommendation for classification
</pre>

====================================================================== Einde pagina 1 =================================================================

<br><br>====================================================================== Pagina 2 ======================================================================

<pre>Aan de Staatssecretaris van Sociale Zaken en Werkgelegenheid
Onderwerp       :  aanbieding advies
Uw kenmerk      :  DGV/BMO-U-932542
Ons kenmerk     :  U 949/AB/jt/543-Z2
Bijlagen        :  1
Datum           :  3 mei 2000
Mijnheer de Staatssecretaris,
Bij brief van 3 december 1993, nr. DGV/BMO-U-932542, verzocht de Staatssecretaris
van Welzijn, Volksgezondheid en Cultuur namens de Minister van Sociale Zaken en
Werkgelegenheid om naast het afleiden van gezondheidskundige advieswaarden ook te
adviseren over andere onderwerpen ten behoeve van de bescherming van beroepsmatig
aan stoffen blootgestelde personen. In 1995 heeft de Staatssecretaris van Sociale Zaken
en Werkgelegenheid besloten tot het opstellen van een zogenaamde niet-limitatieve repro-
tox-lijst. Op deze lijst komen stoffen die volgens de richtlijnen van de Europese Unie in-
gedeeld moeten worden in categorie 1 of 2 wat betreft effecten op de voortplanting. De
Gezondheidsraad is verzocht om voor stoffen een classificatie volgens de EU-criteria
voor te stellen.
In 1996 heb ik hiervoor de Commissie Reproductietoxische stoffen ingesteld.
Hierbij bied ik u - gehoord de Beraadsgroep Gezondheid en Omgeving - de publikatie
van de commissie aan over chloroform. Deze publikatie heb ik heden ter kennisname aan
de Minister van Volksgezondheid Welzijn en Sport en aan de Minister van
Volkshuisvesting, Ruimtelijke Ordening en Milieubeheer gestuurd.
Hoogachtend,
w.g.
prof. dr JJ Sixma
</pre>

====================================================================== Einde pagina 2 =================================================================

<br><br>====================================================================== Pagina 3 ======================================================================

<pre>Chloroform
Evaluation of the effects on reproduction, recommendation for classification
Committee for Compounds toxic to reproduction,
a committee of the Health Council of the Netherlands
to
the Minister and State Secretary of Social Affairs and Employment
No. 2000/07OSH, The Hague, 3 May 2000
</pre>

====================================================================== Einde pagina 3 =================================================================

<br><br>====================================================================== Pagina 4 ======================================================================

<pre>Preferred citation:
Health Council of the Netherlands: Committee for Compounds toxic to reproduction.
Chloroform; Evaluation of the effects on reproduction, recommendation for classifica-
tion. The Hague: Health Council of the Netherlands, 2000; publication no.
2000/07OSH.
all rights reserved
ISBN: 90-5549-312-0
</pre>

====================================================================== Einde pagina 4 =================================================================

<br><br>====================================================================== Pagina 5 ======================================================================

<pre>    Contents
    Samenvatting 7
    Executive summary 8
1   Scope 9
1.1 Background 9
1.2 Committee and procedure 9
1.3 Additional considerations 10
1.4 Labelling for lactation 11
1.5 Data 12
1.6 Presentation of conclusions 12
1.7 Final remark 12
2   Chloroform 13
2.1 Introduction 13
2.2 Human studies 13
2.3 Animal studies 15
2.4 Conclusion 19
    References 21
    Contents                       5
</pre>

====================================================================== Einde pagina 5 =================================================================

<br><br>====================================================================== Pagina 6 ======================================================================

<pre>  Annexes 24
A The committee 25
B Comments on the public draft 27
C Directive (93/21/EEC) of the European Community 28
D Fertility and developmental toxicity studies 34
E Abbreviations 38
  Contents                                           6
</pre>

====================================================================== Einde pagina 6 =================================================================

<br><br>====================================================================== Pagina 7 ======================================================================

<pre>Samenvatting
Op verzoek van de Minister van Sociale Zaken en Werkgelegenheid beoordeelt de Ge-
zondheidsraad de effecten op de reproductie van stoffen waaraan mensen tijdens de be-
roepsuitoefening kunnen worden blootgesteld. De Commissie Reproductietoxische
stoffen, een commissie van de Raad, adviseert een classificatie van reproductietoxische
stoffen volgens Richtlijn 93/21/EEC van de Europese Unie. In het voorliggende rapport
heeft de commissie chloroform onder de loep genomen.
De aanbevelingen van de commissie zijn:
    Voor effecten op de fertiliteit, meent de commissie dat er onvoldoende gegevens be-
    schikbaar zijn. Zij adviseert daarom chloroform niet te classificeren.
    Voor ontwikkelingsstoornissen, adviseert de commissie om chloroform in categorie 2
    (Stoffen die dienen te worden beschouwd alsof zij bij de mens ontwikkelingsstoor-
    nissen veroorzaken) te classificeren en met R61 (kan het ongeboren kind schaden)
    te kenmerken.
    Voor effecten tijdens lactatie, adviseert de commissie om chloroform niet te kenmer-
    ken wegens gebrek aan bewijs.
Samenvatting                                                                             7
</pre>

====================================================================== Einde pagina 7 =================================================================

<br><br>====================================================================== Pagina 8 ======================================================================

<pre>Executive summary
On request of the Minister of Social Affairs and Employment, the Health Council of the
Netherlands evaluates the effects on the reproduction of substances at the workplace.
The Health Council’s Committee for Compounds Toxic to Reproduction recommends to
classify compounds toxic to reproduction according to the Directive 93/21/EEC of the
European Union. In the present report the committee has reviewed chloroform.
The committee’s recommendations are:
    For effects on fertility, the committee recommends no classification of chloroform
    due to a lack of appropriate data.
    For developmental toxicity, the committee recommends to classify chloroform in ca-
    tegory 2 (Substances which should be regarded as if they developmental toxicity to
    humans) and to label chloroform with R61 (may cause harm to the unborn child).
    For effects during lactation, the committee is of the opinion that due to the lack of
    appropriate data chloroform should not be labelled with R64.
Executive summary                                                                         8
</pre>

====================================================================== Einde pagina 8 =================================================================

<br><br>====================================================================== Pagina 9 ======================================================================

<pre>Chapter 1
        Scope
1.1     Background
        As a result of the Dutch regulations on registration of compounds toxic to reproduction
        that came into force on 1 April 1995, the Minister of Social Affairs and Employment re-
        quested the Health Council of the Netherlands to classify compounds toxic to reproduc-
        tion. The classification is performed according to the guidelines of the European Union
        (Directive 93/21/EEG) by the Health Council’s Committee for Compounds Toxic to Re-
        production. The committee’s advice on the classification will be applied by the Ministry
        of Social Affairs and Employment to extend the existing list of compounds classified as
        toxic to reproduction (class 1 and 2) of the European Union.
1.2     Committee and procedure
        The present document contains the classification of chloroform by the Health Council’s
        Committee for Compounds Toxic to Reproduction. The members of the committee are
        listed in annex A. The first draft of this report was prepared by Ms ir IDH Waalkens-
        Berendsen, Mr drs WG Blijleven (deceased January 1997) and Ms dr AE Smits-van
        Prooije at the Department of Neurotoxicology and Reproduction Toxicology of the TNO
        Nutrition and Food Research Institute, Zeist, The Netherlands, by contract with the Mi-
        nistry of Social Affairs and Employment. The classification is based on the evaluation of
        published human and animal studies concerning adverse effects with respect to fertility
        and development and lactation of the above mentioned compound.
        Scope                                                                                     9
</pre>

====================================================================== Einde pagina 9 =================================================================

<br><br>====================================================================== Pagina 10 ======================================================================

<pre>    Classification and labelling was performed according to the guidelines of the European
    Community listed in annex C.
     Classification for fertility and development:
     Category 1          Substances known to impair fertility in humans (R60)
                         Substances known to cause developmental toxicity in humans (R61)
     Category 2          Substances which should be regarded as if they impair fertility in humans (R60)
                         Substances which should be regarded as if they cause developmental toxicity in hu-
                         mans (R61)
     Category 3          Substances which cause concern for human fertility (R62)
                         Substances which cause concern for humans owing to possible developmental toxic
                         effects (R63)
     No classification for effects on fertility or development
     Labelling for lactation:
                         May cause harm to breastfed babies (R64)
                         No labelling for lactation
    In April 1999, the President of the Health Council released a draft of the report for pu-
    blic review. The individuals and organisations that commented on the draft report are lis-
    ted in annex B. The committee has taken these comments into account in deciding on the
    final version of the report.
1.3 Additional considerations
    The classification of compounds toxic to reproduction on the basis of the Directive
    93/21/EEC is ultimately dependent on an integrated assessment of the nature of all pa-
    rental and developmental effects observed, their specificity and adversity, and the dosa-
    ges at which the various effects occur. The directive necessarily leaves room for
    interpretation, dependent on the specific data set under consideration. In the process of
    using the directive, the committee has agreed upon a number of additional considera-
    tions.
         If there is sufficient evidence to establish a causal relationship between human expo-
         sure to the substance and impaired fertility or subsequent developmental toxic ef-
         fects in the progeny, the compound will be classified in category 1, irrespective the
         general toxic effects (see Annex C, 4.2.3.1 category 1).
         Adverse effects in a reproductive or developmental study, in the absence of data on
         parental toxicity, occurring at dose levels which cause severe toxicity in other stu-
         dies, need not necessarily lead to a category 2 classification.
    Scope                                                                                                   10
</pre>

====================================================================== Einde pagina 10 =================================================================

<br><br>====================================================================== Pagina 11 ======================================================================

<pre>         If, after prenatal exposure, small reversible changes in foetal growth and in skeletal
         development (e.g. wavy ribs, short rib XIII, incomplete ossification) in offspring oc-
         cur in a higher incidence than in the control group in the absence of maternal effects,
         the substance will be classified in category 3 for developmental toxicity. If these ef-
         fects occur in the presence of maternal toxicity, they will be considered as a conse-
         quence of this and therefore the substance will not be classified for developmental
         toxicity (see Annex C, 4.2.3.3 developmental toxicity final paragraph).
         Clear adverse reproductive effects will not be disregarded on the basis of reversibili-
         ty per se.
         Effects on sex organs in a general toxicity study (e.g. in a subchronic or chronic
         toxicity study) may warrant classification for fertility.
         The committee not only uses guideline studies (studies performed according to
         OECD standard protocols*) for the classification of compounds, but non-guideline
         studies are taken into consideration as well.
1.4 Labelling for lactation
    The recommendation for labelling substances for effects during lactation is also based on
    Directive 93/21/EEC. The Directive defines that substances which are absorbed by wo-
    men and may interfere with lactation or which may be present (including metabolites) in
    breast milk in amounts sufficient to cause concern for the health of a breastfed child,
    should be labelled with R64. Unlike the classification of substances for fertility and de-
    velopmental effects, which is based on a hazard identification only (largely independent
    of dosage), the labelling for effects during lactation is based on a risk characterization
    and therefore also includes consideration of the level of exposure of the breastfed child.
         Consequently, a substance should be labelled for effects during lactation when it is
    likely that the substance would be present in breast milk in potentially toxic levels. The
    committee considers a compound as potentially toxic to the breastfed child when exposu-
    re to this compound via the milk results in an intake exceeding an exposure limit for the
    general population, eg the acceptable daily intake (ADI).
*   Organisation for Economic Cooperation and Development
    Scope                                                                                        11
</pre>

====================================================================== Einde pagina 11 =================================================================

<br><br>====================================================================== Pagina 12 ======================================================================

<pre>1.5 Data
    Literature searches were conducted in the on-line databases Toxline and Medline, star-
    ting from 1966 up to 1995. Literature was selected primarily on the basis of the ab-
    stracts. Publications cited in the selected articles, but not selected during the primary
    search, were reviewed if considered appropriate by the committee. In addition, hand-
    books and a collection of most recent reviews were consulted. References are divided in
    literature cited and literature consulted but not cited. Before finalising the public draft
    report, the committee performed an additional literature search in Medline and Toxline
    for the period 1995 to 1998. The results of this search were no reason for the committee
    to adjust the recommendations.
         The committee chose to describe human studies in the text, starting with review arti-
    cles. Of each study the quality of the study design (performed according to international-
    ly acknowledged guidelines) and the quality of documentation are indicated.
         Animal data are summarised in tables in annex D. Special attention was paid to re-
    versibility of observed effects and data on general toxicity and maternal toxicity.
1.6 Presentation of conclusions
    The classification is given with key effects, species and references specified. In case a
    substance is not classified as toxic to reproduction, one of two reasons is given:
         Lack of appropriate data preclude assessment of the compound for reproductive
         toxicity.
         Sufficient data show that no classification for toxic to reproduction is indicated.
1.7 Final remark
    The classification of compounds is based on hazard evaluation* only, which is one of a
    series of elements guiding the risk evaluation process. The committee emphasises that
    for derivation of health based occupational exposure limits these classifications should
    be placed in a wider context. For a comprehensive risk evaluation, hazard evaluation
    should be combined with dose-response assessment, human risk characterization, human
    exposure assessment and recommendations of other organisations.
*   for definitions see Tox95
    Scope                                                                                       12
</pre>

====================================================================== Einde pagina 12 =================================================================

<br><br>====================================================================== Pagina 13 ======================================================================

<pre>Chapter 2
        Chloroform
2.1     Introduction
        Chloroform is a colourless and odourless liquid.
         Name           :   chloroform
         CAS-no         :   67-66-3
         Synonym        :   trichloromethane
         Use            :   as solvent for fats, oils, waxes, resins; as cleaning agent; in fire extinguishers; as
                            flavouring substance and preservative; in manufacturing of dyes, drugs and pesti-
                            cides
         Mol weight     :   119.39
         Chem formula   :   CHCl3
2.2     Human studies
        Fertility
        The pregnancy outcome of female workers in the pharmaceutical industry was
        investigated in a case-referent study (Tas90): exposure to organic solvents was
        associated with the occurrence of spontaneous abortion in a dose-related manner.
        Chloroform was among these solvents, but individual relationships could not be proven.
        Chloroform                                                                                                 13
</pre>

====================================================================== Einde pagina 13 =================================================================

<br><br>====================================================================== Pagina 14 ======================================================================

<pre>     Sullivan et al. (Sul93) concluded that data available are inadequate to assess the
potential of chloroform to induce spontaneous abortions or congenital malformations.
Developmental toxicity
Cardiac defects, neural tube defects, oral clefts and very low birth weight were studied in
a case-control study in 4 counties in northern New Jersey (Bov92). Statistically
significant associations were found between neural tube defects and concentrations of
total trihalomethanes (TTHM) higher than 80 ppb with an odds ratio (OR) of 4.5 and
concentrations of total trihalomethanes (TTHM) higher than 15 ppb with an OR of 3.8.
For oral clefts, cardiac defects and low birth weight no statistically significant
associations were found with TTHM. Mothers of cases and controls were interviewed by
telephone; a total of 563 mothers were interviewed.
     After adjustment for maternal age, parity, adequacy of prenatal care, marital status,
education and maternal smoking, Kramer et al. (Kra92) found an association between
chloroform levels higher than 10 ppb and an increased prevalence of smaller infants (for
the time of gestation) of non-hispanic, white women from Iowa. The major limitations of
this study involve the ascertainment and classification of exposures to trihalomethanes.
     Bove et al. (Bov95) conducted a cross-sectional study using environmental and birth
outcome data bases in 4 counties in northern New Jersey where some water supplies
were contaminated with total trihalomethane (TTHM). A total of 80,938 live births and
594 foetal deaths were studied. Odds ratios higher than 1.50 were found for the
following: TTHM with smaller infants (for the time of gestation), central nervous system
defects and oral clefts. TTHM levels greater than 100 ppb reduced birth weight among
term births by 70.4 g. The authors stated that this study cannot resolve whether drinking
water contaminants caused the adverse birth outcome.
     Savitz et al. (Sav95) evaluated the risk associated with water source, amount of
water ingestion and trihalothanes (THM) concentration using data from-a case-control
study of miscarriage, preterm delivery and low-birth weight in North Carolina. The data
from the study do not indicate a strong association between chlorination by-products and
adverse pregnancy outcome, with possible exception of an increased risk of miscarriage
in the highest sextile of THM concentration (adjusted odds ratio= 2.8, 95% interval =
1.1-2.7).
     Waller et al. (Wal98) examined the exposure to trihalothanes and spontaneous
abortion in a prospective study of 5,144 pregnant women in California. Of the four
individual trihalothanes, only high bromodichloro methane exposure (consumption of 5
glasses per day of cold tap water containing 18 µg per litre bromodichloro methane) was
associated with spontaneous abortion.
Chloroform                                                                                  14
</pre>

====================================================================== Einde pagina 14 =================================================================

<br><br>====================================================================== Pagina 15 ======================================================================

<pre>    Reiff et al criticized several of the above mentioned studies because of the lack of
    exposure data. They were of the opinion that the positive findings should be interpreted
    cautiously (Rei96). The committee concluded that these studies concern mixtures of
    chlorination products and/or that human exposures were derived from indirect estimates.
    This limits the establishment of a causal relation between (the single compound)
    chloroform and the birth defects that were found. Therefore, the committee is of the
    opinion that the studies of Kramer et al, Bove et al, Savitz et al and Waller et al do not
    provide sufficient evidence to propose a classification for chloroform.
    Lactation
    Lechner et al. 1988 compared the concentrations of chloroform, carbon tetrachloride and
    tetrachloroethylene in breast milk of 13 mothers from the surroundings of Innsbruck
    with breast milk of 20 mothers from Linz, a more industrialized area in Austria (Lec88).
    No elevation in chlorocarbon levels was detected in either group.
         Fisher et al. 1997 studied the human blood/air and milk/air partition coefficient in
    blood and milk samples donated by lactating women (n=9) (Fis97). The objective of this
    study was to evaluate the potential chemical exposure of a nursing infant by ingestion of
    contaminated milk from a mother who was occupationally exposed to vapours. To esti-
    mate infants’ exposure, a generic human pharmacokinetic (PB-PK) lactation model was
    developed. The model was based on a 8-hour exposure of the mother to a constant
    vapour concentration equal to the threshold limit value for chloroform (10 ppm) in
    drinking water. The experimentally determined blood/air and milk/air partition
    coefficient values were used in the PB-PK lactation model. The predicted amount of
    chloroform ingested by a nursing infant over a 24-hour period was 0.043 mg. However,
    this model has not been validated yet and the relevance of this exposure level to the deve-
    lopment of the human infant is unknown.
2.3 Animal studies
    Tables 1 and 2 (Annex D) summarize the effects of chloroform on fertility and
    development in experimental animals.
    Fertility
    Gulati et al. (Gul97, NTP summary) evaluated the effects of chloroform on fertility in
    CD-1 mice by using a continuous breeding protocol. Chloroform was administered by
    oral gavage using corn oil as a vehicle. Based on a 14-day dose-response study 8, 20,
    and 50 mg/kg body weight were chosen to test the effect on fertility and development.
    Chloroform                                                                                  15
</pre>

====================================================================== Einde pagina 15 =================================================================

<br><br>====================================================================== Pagina 16 ======================================================================

<pre>Based on a reference analysis of representative aliquots, it was estimated that the actual
doses received were 6.6, 16, and 41 mg/kg bw in the low, mid and high dose groups,
respectively. Both male and female mice (20 pairs per treatment group, 40 pairs for
control animals) were dosed daily for 7 days prior to and during a 98-day cohabitation
period. At the high dose chloroform treatment, body weight was not clearly affected in
the parental generation. No effects on fertility were observed.
     In 1992, The Nordic Council of Ministers group and the Nordic chemicals control
group (Dan92) reviewed the available literature on the toxicology and reproductive
hazards of chloroform. It was concluded that data concerning effects of chloroform on
fertility and gonadal function in humans, as well as effects on fertility in animals, are
either absent or inadequate for the evaluation of effects on fertility and gonadal function.
Developmental toxicity
Inhalation studies
Schwetz et al. (Sch74) exposed Sprague-Dawley rats to the subanaesthetic levels of 0,
147, 490 or 1470 mg chloroform/m3 (0, 30, 100 or 300 ppm) for 7 hours per day on
days 6 to 15 of gestation. Exposure to chloroform caused a decrease in the conception
rate, a high incidence of foetal resorption (300 ppm), and a retarded foetal development
(all dose groups). Examination of the foetuses of dams exposed to 100 ppm revealed a
significant incidence of acaudia (absence of tail) or short tails, imperforate anus,
subcutaneous edema, missing ribs and delayed ossification of sternebrae. The number of
foetuses in the 300 ppm group was too low for a meaningful statistical analysis.
Maternal toxicity was observed in the 100 and 300 ppm groups as concluded from
anorexia and changes in absolute and/or relative liver weights. At 30 ppm, a slight
reduction of maternal body weight gain was observed. True terata (acaudia and
imperforate anus) were observed in foetuses of rats exposed to 100 ppm chloroform.
These untoward effects were not attributable to anorexia, since the same degree of
starvation without exposure to chloroform was not embryo- or foetotoxic. In order to
determine whether anorexia was responsible for the embryonic and foetal effects, a
second ‘starved’ control group was included in the experiment. The decrease of foetal
body weight was not associated with developmental effects in this group.
     Dilley et al. (Dil77) exposed pregnant rats (number unknown) daily to 20.1 g/m3
chloroform during days 7 to 14 of gestation. All animals were sacrificed on day 20 of
gestation, and the dams and foetuses were examined for gross changes. Chloroform
caused increased foetal mortality and decreased foetal weight gain, but no teratogenic
effects. Possible maternal toxicity was not described.
Chloroform                                                                                   16
</pre>

====================================================================== Einde pagina 16 =================================================================

<br><br>====================================================================== Pagina 17 ======================================================================

<pre>     Murray et al. (Mur79) exposed mice (number unknown) to 0 or 147 mg/m3 of
chloroform for 7 hours/day from day 1 to 8, 6 to 16 or 8 to 16 of gestation. Maternal
toxicity was observed among the dams exposed to chloroform from day 6-16 of
gestation as a slight decrease in body weight gain, and as significant decrease in body
weight gain among the dams exposed from day 1 to 8 and from day 8 to 16. The ability
of the females to maintain pregnancy was significantly decreased after exposure to
chloroform from day 1 to 8 or 6 to 16 of gestation. A significant increase in the mean
number of resorptions per litter was observed among the mice exposed from day 1 to 8.
Mean foetal body weight and crown-rump length were decreased significantly among the
offspring of mice inhaling chloroform from day 1 to 8 or 8 to 16 of gestation. Cleft
palate occurred significantly more often among the litters of mice exposed to chloroform
from gestation day 8 to 16. Compared to the control group, the incidence of delayed
ossification of skull bones was significantly increased among each of the experimental
groups. In addition, delayed ossification of the sternebrae occurred significantly more
often among the litters of dams that inhaled chloroform from day 1 to 8 or from day 8 to
16, but not from day 6 to 16 of gestation.
     Baeder and Hofmann (1988) exposed mated female Wistar rats by inhalation to 0,
30, 100 or 300 ppm (0, 147, 490 or 1470 mg/m3) from gestation day 7 to 16 during 7
hours per day (Bae88). In the 0, 30, 100 and 300 ppm treated groups 0, 2, 3 and 8 dams
with resorptions only were observed. Foetal weight was decreased in all chloroform
treated groups. Decreased maternal body weight and food intake was observed in the
100 and 300 ppm groups.
     In a supplementary study, Baeder and Hofmann (1991) exposed mated female
Wistar rats by inhalation to 0, 3, 10 or 30 ppm (0, 14.7, 49 or 147 mg/m3) from
gestation day 7 to 16 during 7 hours per day (Bae91). The percentage of small foetuses
was statistically significantly increased in the 10 and 30 ppm groups. In the 30 ppm
group 1 dam with resorptions only was observed. In this group a slightly reduced food
intake was observed as well.
Oral gavage studies
The effect of orally administered chloroform on embryonic and foetal development in the
rat was evaluated by Thompson et al. (Tho74). Rats received doses of 0, 20, 50 or 126
mg/kg bw/day on days 6-16 of gestation. Body weight gain of dams administered 50
mg/kg bw/day was significantly lower in comparison with controls, and dams from the
126 mg/kg bw/day lost weight. Food consumption was significantly decreased in the 126
mg/kg bw/day group. At 126 mg/kg bw/day statistically significantly decreased body
weight of foetuses was observed after administration of chloroform. Teratogenic effects
were not observed.
Chloroform                                                                               17
</pre>

====================================================================== Einde pagina 17 =================================================================

<br><br>====================================================================== Pagina 18 ======================================================================

<pre>         Ruddick et al. (Rud83) administered chloroform by gavage to Sprague-Dawley rats
    from day 6 to 15 of gestation. Chloroform was administered at levels of 100, 200 and
    400 mg/kg. Maternal weight gain was depressed in all groups receiving chloroform.
    Chloroform administration caused decreased maternal haemoglobin and haematocrit
    values at all dose levels and also produced increased serum inorganic phosphorus and
    cholesterol at the highest dose. Liver enlargement was observed at all dose levels of
    chloroform. Evidence of a foetotoxic response (not specified) was observed with
    chloroform, but teratogenic effects were not observed.
         The effect of orally administered chloroform on embryonic and foetal development
    in the rabbit was evaluated by Thompson et al. (Tho74). Rabbits received doses of 0,
    20, 35 or 50 mg/kg bw/day on days 6-19 of gestation. Body weight gain of dams
    administered 50 mg/kg bw/day was significantly decreased in comparison to controls.
    Foetotoxic or teratogenic effects were not observed.
         Gulati et al. (Gul97, only NTP summary) evaluated the effect of chloroform on
    fertility and reproduction in the first and second generation of CD-1 mice by use of a
    continuous breeding protocol at levels of 8, 20, and 50 mg/kg body weight (actual doses
    were 6.6, 16, and 41 mg/kg bw in the low, mid and high dose groups, respectively). Both
    male and female mice (20 pairs per treatment group, 40 pairs for control animals) were
    dosed daily for 7 days prior to and during a 98-day co-habitation period. The offspring
    from the control and from the high dose group, the F1 generation, was also evaluated. In
    the high dose chloroform group, no effects on body weight of either the parents or the
    offspring were observed. F1 generation males in the high dose group showed significantly
    increased epididymal weights. However, the epididymal sperm motility, count and
    morphology were not affected. F1 females in the high dose group showed increased liver
    weight and there were signs of hepatocellular degeneration.
2.4 Conclusion
    No relevant data were found concerning the effects of chloroform on human or animal
    fertility. The potential effect of chloroform as a cause of abortion in humans could not
    be proven (Tas90). In conclusion, a lack of appropriate data precludes the assessment of
    chloroform for effects on fertility. Therefore chloroform should not be classified with
    respect to effects on fertility.
    The data concerning the developmental effects in humans (Bov92, Kra92, Bov95,
    Sav95, Wal98) were insufficient to classify chloroform.
         In two inhalatory studies with animals, developmental effects were observed in rats
    and mice, in the presence of maternal toxicity (Sch74; Mur79). These developmental
    effects (eg acaudia and imperforate anus) were considered not secondary to the maternal
    Chloroform                                                                               18
</pre>

====================================================================== Einde pagina 18 =================================================================

<br><br>====================================================================== Pagina 19 ======================================================================

<pre>toxicity. In addition, Baeder and Hofmann (Bae88 and Bae91) observed an increased
number of dams with resorptions only, a decreased foetal weight and an increased
number of small foetuses at dose levels at which no or only slight maternal toxicity was
observed.
     In conclusion, the committee is of the opinion that the effects on development were
specific (acaudia and imperforate anus in rat (Sch74) and pregnancy maintenance in
mice (Mur79)) and could not be explained by maternal toxicity. The studies of Baeder
and Hofmann (Bae88 and Bae91) support the findings of Murray and Schwetz (Muy79;
Sch74) that inhalatory exposure to chloroform causes an increased number of resorp-
tions.
     Therefore, in view of the animal data, the committee recommends to classify
chloroform in category 2, substances which should be regarded as if they cause
developmental toxicity in humans. Chloroform should be labelled with R61 (may cause
harm to the unborn child).
No data were available concerning the transfer of chloroform to human breast milk, but
due to its physicochemical properties (Fis97) it may be assumed that chloroform passes
to breast milk. However, the committee is of the opinion that this assumption is not
sufficient for labelling chloroform for lactation. No data were available concerning
effects during lactation.
     In conclusion, a lack of appropriate data precludes the labelling of chloroform for
effects during lactation. Therefore chloroform is not labelled with respect to effects
during lactation.
Proposed classification for effects on fertility
A lack of appropriate data preclude the assessment of chloroform for fertility.
Proposed classification for developmental toxicity
Category 2, R 61.
Proposed labelling for effects during lactation
A lack of appropriate data precludes the assessment of chloroform for effects during lac-
tation.
Chloroform                                                                                19
</pre>

====================================================================== Einde pagina 19 =================================================================

<br><br>====================================================================== Pagina 20 ======================================================================

<pre>For the committee,
The Hague, 3 May 2000
dr ASAM van der Burght, dr BJ Blaauboer,
scientific secretary    chairman
Chloroform                               20
</pre>

====================================================================== Einde pagina 20 =================================================================

<br><br>====================================================================== Pagina 21 ======================================================================

<pre>      References
Bae88 Baeder Ch, Hofmann Th. Chloroform. Prüfung auf embryotoxische Wirkung an Wistar-Ratten bei inhala-
      tiver Verabreichung. Hoechst Bericht Nr. 88.0609 13. Mai 1988 Seite 1-68. (Confidential report submit-
      ted to the committee by Elf Atochem and to be consulted at the Health Council of the Netherlands, Den
      Haag)
Bae91 Baeder C, Hofmann T. Chloroform. Supplementary Inhalation embryotoxicity study in Wistar rats.
      Hoechst Report No. 91.0902 September 12. 1991 Page 1-38 (39-365; individual data not available),
      (Confidential report submitted to the committee by Elf Atochem and to be consulted at the Health Coun-
      cil of the Netherlands, Den Haag)
Bov92 Bove FJ, Fulcomer MC, Klotz JB, et al. Public drinking water contamination and birth weight and selec-
      ted birth defects: A case control study. Trenton NJ: New Yersey Department of Health 1992.
Bov95 Bove FJ, Fulcomer MC, Klotz JB, et al. Public drinking water contamination and birth outcomes.Am J
      Epidemiol 1995; 141: 850-862.
Dan92 Danielsson BR. Effects on reproduction of chloroform (trichloromethane). Nordic chemicals control
      group. Keml Report 1992; 7/92: 9-40 (only abstract available).
Dil77 Dilley JV, Chernoff N, Kay D, et al. Inhalation teratology studies of five chemicals in rats. Toxicol. Appl.
      Pharmacol. 1977; 41: 196 (conference abstract).
Fis97 Fisher J, Mahle D, Bankston L, et al. Lactational transfer of volatile chemicals in breast milk. Am Ind
      Hyg Ass J 1997; 58:425-431.
Gul97 Gulati DK, Hope E, Mounce RC, et al. Chloroform NTIS#PB89 14868639/AS Environ. Health Perspec.
      1997; 105, Suppl 1:285-286 (summary only).
Kra92 Kramer MD, Lynch CF, Isacson P, et al. The association of waterborne chloroform with intrauterine
      growth retardation. Epidemiology 1992; 3:407-413.
      References                                                                                                   21
</pre>

====================================================================== Einde pagina 21 =================================================================

<br><br>====================================================================== Pagina 22 ======================================================================

<pre>Lec88 Lechner W, Bonn G, Thews G, et al. Chlorierte Kohlwasserstoffe in der Muttermilch. Wien Klin Woch
      1988; 100(18); 622-624.
Mur79 Murray FJ, Schwetz BA, McBride JG, et al. Toxicity of inhaled chloroform in pregnant mice and their
      offspring. Toxicol. Appl. Pharmacol 1979; 50: 515-22.
Rei96 Reiff JS, Hatch MC, Bracken M, et al. Reproductive and developmental effects of disinfection by-pro-
      ducts in drinking water. Environ. Health Perspec. 1996; 10: 1056-1061.
Rud83 Ruddick JV, Villeneuve DC, Chu I. A teratological assessment of four trihalomethanes in the rat. J. Envi-
      ron. Sci. Health 1983; B18(3) 333-49.
Sav95 Savitz DA, Andrews KW, Pastore LM. Drinking water and pregnancy outcome in Central North Caroli-
      na: source, amount, and trihalothane levels. Environ. Health Perspec. 1995; 103: 592-596.
Sch74 Schwetz BA, Leong BKJ, Gehring PJ. Embryo and fetotoxicity of inhaled chloroform in rats. Toxicol
      Appl Pharmacol 1974; 28: 442-51.
Sul93 Sullivan FM, Watkins WJ, van der Venne MTh. Reproductive toxicity 1993; 1: 81-92. Summary reviews
      of the scientific evidence. Commission of the European Communities.
Tas90 Taskinen HK. Effects of parental occupational exposures on sponteneous abortion and congenital malfor-
      mation. Scand J Work Environ Health 1990; 16: 297-314.
Tho74 Thompson DJ, Warner SD, Robinson VB. Teratology studies on orally administered chloroform in the rat
      and rabbit. Toxicol Appl Pharmacol 1974; 29: 348-57.
Tox95 Niesink RJM, de Vries J, Hollinger MA, eds, Toxicology, Principles and Applications, Boca Raton: CRC
      Press, 1995:385
Wal98 Waller K, Swan SH, DeLorenze G, et al. Trihalomethanes in drinking water and spontaneous abortion.
      Epidemiology 1998; 9: 134-140.
      Literature consulted but not used
Cor90 Corley RA, Mendrala AL, Smith FA, et al. Development of a physiologically based pharmacokinetic mo-
      del for chloroform. Toxicol Appl Pharmacol 1990; 103:512-527.
EPA92 EPA/OTS. Initial submission: inhalation embryotoxicity study with chloroform in Wistar rats. EPA/OTS
      Doc #88-920005781 1992 (final report EPA/OTS Doc #88-920001208) (only abstract available).
Gul89 Gulati DK, Hope E, Mounce RC, et al. Chloroform Reproduction and Fertility Assessment in CD-1 Mice
      When Administered by Gavage. Govt Reports Announcements & Index (GRA&I), Issue 10 1989 (only
      abstract available).
Mer84 Mercier M, Lans M, de Gerlache J. Mutagenicity, carcinogenicity and teratogenicity of halogenated hy-
      drocarbon solvents. Mutagen Carcinog Teratog Ind Pollut 1984; 281-324.
Nor92 Nordic Council of Ministers. Effects on reproduction of chloroform and 1,2-dibromo-3-chloropropane.
      1992; Swedish National Chemicals Inspectorate, P.O. Box 1384, 171 27 Solna, Sweden.
Smi86 Smith MK, Zenick H, George EL. Reproductive toxicology of disinfection by-products. Environmental
      Health Perspectives 1986; 69: 177-82.
      References                                                                                                22
</pre>

====================================================================== Einde pagina 22 =================================================================

<br><br>====================================================================== Pagina 23 ======================================================================

<pre>WGD87 WGD. Rapport inzake grenswaarde chloroform. Gezondheidskundig advies van de Werkgroep van Des-
      kundigen ter vaststelling van MAC-waarden. 1987, Directoraat-Generaal van de Arbeid, RA 7/87.
      References                                                                                     23
</pre>

====================================================================== Einde pagina 23 =================================================================

<br><br>====================================================================== Pagina 24 ======================================================================

<pre>A The committee
B Comments on the public draft
C Directive (93/21/EEG) of the European Community
D Fertility and developmental toxicity studies
E Abbreviations
  Annexes
                                                  24
</pre>

====================================================================== Einde pagina 24 =================================================================

<br><br>====================================================================== Pagina 25 ======================================================================

<pre>Annex A
      The committee
         BJ Blaauboer, chairman
         Toxicologist; Research Institute of Toxicology, Utrecht
         JN van den Anker
         Professor of pediatrics and Neonatology; Erasmus University, Rotterdam
         AM Bongers, advisor
         Ministry of Social Affairs and Employment, The Hague
         HFP Joosten
         Toxicologist; NV Organon, Department of Toxicology and Drug Disposition, Oss
         D Lindhout
         Professor of Clinical Genetics/Teratology; Erasmus University, Rotterdam
         JHJ Copius Peereboom-Stegeman
         Toxicologist; Catholic University Nijmegen, Nijmegen
         AH Piersma
         Reproductive toxicologist; National Institute of Public Health and the Environment,
         Bilthoven
         A Stijkel
         Toxicologist; Environmental Awareness Foundation, ’s-Graveland
         PJJM Weterings
         Toxicologist; Weterings Consultancy BV, Rosmalen
         ASAM van der Burght, scientific secretary
         Health Council of the Netherlands, The Hague
      The committee                                                                          25
</pre>

====================================================================== Einde pagina 25 =================================================================

<br><br>====================================================================== Pagina 26 ======================================================================

<pre>The first draft of the present document was prepared by AE Smits-van Prooije, WG
Blijleven and IDH Waalkens-Berendsen, from the TNO Nutrition and Food Research In-
stitute in Zeist, by contract with the Ministry of Social Affairs and Employment.
Secretarial assistance: E Vandenbussche-Parméus.
Lay-out: J van Kan.
The committee                                                                      26
</pre>

====================================================================== Einde pagina 26 =================================================================

<br><br>====================================================================== Pagina 27 ======================================================================

<pre>Annex B
      Comments on the public draft
      A draft of the present report was released in 1999 for public review. The following orga-
      nisations and persons have commented on the draft document:
          RJ Millischer, Elf Atochem SA, France.
      Comments on the public draft                                                              27
</pre>

====================================================================== Einde pagina 27 =================================================================

<br><br>====================================================================== Pagina 28 ======================================================================

<pre>Annex C
      Directive (93/21/EEC) of the European
      Community
      4.2.3         Substances toxic to reproduction
      4.2.3.1       For the purposes of classification and labelling and having regard to the present
                    state of knowledge, such substances are divided into 3 categories:
      Category 1:
      Substances known to impair fertility in humans
      There is sufficient evidence to establish a causal relationship between human exposure to the substance
      and impaired fertility.
      Substances known to cause developmental toxicity in humans
      There is sufficient evidence to establish a causal relationship between human exposure to the substance
      and subsequent developmental toxic effects in the progeny.
      Category 2:
      Substances which should be regarded as if they impair fertility in humans:
      Directive (93/21/EEC) of the European Community                                                         28
</pre>

====================================================================== Einde pagina 28 =================================================================

<br><br>====================================================================== Pagina 29 ======================================================================

<pre>There is sufficient evidence to provide a strong presumption that human exposure to the substance may
result in impaired fertility on the basis of:
     Clear evidence in animal studies of impaired fertility in the absence of toxic effects, or, evidence of
     impaired fertility occurring at around the same dose levels as other toxic effects but which is not a
     secondary non-specific consequence of the other toxic effects.
     Other relevant information.
Substances which should be regarded if they cause developmental toxicity to humans:
There is sufficient evidence to provide a strong presumption that human exposure to the substance may
result in developmental toxicity, generally on the basis of:
     Clear resuts in appropriate animal studies where effects have been observed in the absence of signs
     of marked maternal toxicity, or at around the same dose levels as other toxic effects but which are
     not a secondary non-specific consequence of the other toxic effects.
     Other relevant information.
Category 3:
Substances which cause concern for human fertility:
Generally on the basis of:
     Results in appropriate animal studies which provide sufficient evidence to cause a strong suspicion
     of impaired fertility in the absence of toxic effects, or evidence of impaired fertility occurring at
     around the same dose levels as other toxic effects, but which is not a secondary non-specific conse-
     quence of the other toxic effects, but where the evidence is insufficient to place the substance in Ca-
     tegory 2.
     Other relevant information.
Substances which cause concern for humans owing to possible developmental toxic effects:
Generally on the basis of:
     Results in appropriate animal studies which provide sufficient evidence to cause a strong suspicion
     of developmental toxicity in the absence of signs of marked maternal toxicity, or at around the same
     dose levels as other toxic effects but which are not a secondary non-specific consequence of the
     other toxic effects, but where the evidence is insufficient to place the substance in Category 2.
     Other relevant information.
Directive (93/21/EEC) of the European Community                                                              29
</pre>

====================================================================== Einde pagina 29 =================================================================

<br><br>====================================================================== Pagina 30 ======================================================================

<pre>4.2.3.2       The following symbols and specific risk phrases apply:
Category 1:
For substances that impair fertility in humans:
T; R60: May impair fertility
For substances that cause developmental toxicity:
T; R61: May cause harm to the unborn child
Category 2:
For substances that should be regarded as if they impair fertility in humans:
T; R60: May impair fertility
For substances that should be regarded as if they cause developmental toxicity in humans:
T; R61: May cause harm to the unborn child.
Category 3:
For substances which cause concern for human fertility:
Xn; R62: Possible risk of impaired fertility
For substances which cause concern for humans owing to possible developmental toxic effects:
Xn; R63: Possible risk of harm to the unborn child.
4.2.3.3       Comments regarding the categorisation of substances toxic to reproduction
Reproductive toxicity includes impairment of male and female reproductive functions or capacity and the
induction of non-inheritable harmful effects on the progeny. This may be classified under two main head-
ings of 1) Effects on male or female fertility, 2) Developmental toxicity.
1    Effects on male or female fertility, includes adverse effects on libido, sexual behaviour, any aspect
     of spermatogenesis or oogenesis, or on hormonal activity or physiological response which would in-
Directive (93/21/EEC) of the European Community                                                            30
</pre>

====================================================================== Einde pagina 30 =================================================================

<br><br>====================================================================== Pagina 31 ======================================================================

<pre>      terfere with the capacity to fertilise, fertilisation itself or the development of the fertilised ovum up
      to and including implantation.
2     Developmental toxicity, is taken in its widest sense to include any effect interfering with normal de-
      velopment, both before and after birth. It includes effects induced or manifested prenatally as well
      as those manifested postnatally. This includes embrytoxic/fetotoxic effects such as reduced body
      weight, growth and developmental retardation, organ toxicity, death, abortion, structural defects (te-
      ratogenic effects), functional defects, peri-postnatal defects, and impaired postnatal mental or physi-
      cal development up to and including normal pubertal development.
Classification of chemicals as toxic to reproduction is intended to be used for chemicals which have an
intrinsic or specific property to produce such toxic effects. Chemicals should not be classified as toxic to
reproduction where such effects are solely produced as a non-specific secondary consequence of other
toxic effects. Chemicals of most concern are those which are toxic to reproduction at exposure levels
which do not produce other signs of toxicity.
The placing of a compound in Category 1 for effects on Fertility and/or Developmental Toxicity is done
on the basis of epidemiological data. Placing into Categories 2 or 3 is done primarily on the basis of ani-
mal data. Data from in vitro studies, or studies on avian eggs, are regarded as ‘supportive evidence’ and
would only exceptionally lead to classification in the absence of in vivo data.
In common with most other types of toxic effect, substances demonstrating reproductive toxicity will be
expected to have a threshold below which adverse effects would not be demonstrated. Even when clear
effects have been demonstrated in animal studies the relevance for humans may be doubtful because of
the doses administrated, for example, where effects have been demonstrated only at high doses, or where
marked toxicokinetic differences exist, or the route of administration is inappropriate. For these or simi-
lar reasons it may be that classification in Category 3, or even no classification, will be warranted.
Annex V of the Directive specifies a limit test in the case of substances of low toxicity. If a dose level of
at least 1000 mg/kg orally produces no evidence of effects toxic to reproduction, studies at other dose le-
vels may not be considered necessary. If data are available from studies carried out with doses higher
than the above limit dose, this data must be evaluated together with other relevant data. Under normal
circumstances it is considered that effects seen only at doses in excess of the limit dose would not neces-
sarily lead to classification as Toxic to Reproduction.
Effects on fertility
For the classification of a substance into Category 2 for impaired fertility, there should normally be clear
evidence in one animal species, with supporting evidence on mechanism of action or site of action, or
chemical relationship to other known antifertility agents or other information from humans which would
Directive (93/21/EEC) of the European Community                                                                 31
</pre>

====================================================================== Einde pagina 31 =================================================================

<br><br>====================================================================== Pagina 32 ======================================================================

<pre>lead to the conclusion that effects would be likely to be seen in humans. Where there are studies in only
one species without other relevant supporting evidence then classification in Category 3 may be appropri-
ate.
Since impaired fertility may occur as a non-specific accompaniment to severe generalised toxicity or
where there is severe inanition, classification into Category 2 should only be made where there is eviden-
ce that there is some degree of specificity of toxicity for the reproductive system. If it was demonstrated
that impaired fertility in animal studies was due to failure to mate, then for classification into Category 2,
it would normally be necessary to have evidence on the mechanism of action in order to interpret whether
any adverse effect such as alteration in pattern of hormonal release would be likely to occur in humans.
Developmental toxicity
For classification into Category 2 there should be clear evidence of adverse effects in well conducted stu-
dies in one or more species. Since adverse effects in pregnancy or postnatally may result as a secondary
consequence of maternal toxicity, reduced food or water intake, maternal stress, lack of maternal care,
specific dietary deficiencies, poor animal husbandry, intercurrent infections, and so on, it is important
that the effects observed should occur in well conducted studies and at dose levels which are not associa-
ted with marked maternal toxicity. The route of exposure is also important. In particular, the injection of
irritant material intraperitoneally may result in local damage to the uterus and its contents, and the re-
sults of such studies must be interpreted with caution and on their own would not normally lead to classi-
fication.
Classification into Category 3 is based on similar criteria as for Category 2 but may be used where the
experimental design has deficiencies which make the conclusions less convincing, or where the possibili-
ty that the effects may have been due to non-specific influences such as generalised toxicity cannot be ex-
cluded.
In general, classification in category 3 or no category would be assigned on an ad hoc basis where the
only effects recorded are small changes in the incidences of spontaneous defects, small changes in the
proportions of common variants such as are observed in skeletal examinations, or small differences in
postnatal developmental assessments.
Effects during Lactation
Substances which are classified as toxic to reproduction and which also cause concern due to their effects
on lactation should in addition be labelled with R64 (see criteria in section 3.2.8).
Directive (93/21/EEC) of the European Community                                                                32
</pre>

====================================================================== Einde pagina 32 =================================================================

<br><br>====================================================================== Pagina 33 ======================================================================

<pre>For the purpose of classification, toxic effects on offspring resulting only from exposure via the breast
milk, or toxic effects resulting from direct exposure of children will not be regarded as ‘Toxic to Repro-
duction’, unless such effects result in impaired development of the offspring.
Substances which are not classified as toxic to reproduction but which cause concern due to toxicity
when transferred to the baby during the period of lactation should be labelled with R64 (see criteria in
section 3.2.8). This R-phrase may also be appropriate for substances which affect the quantity or quality
of the milk.
R64 would normally be assigned on the basis of:
a    toxicokinetic studies that would indicate the likelihood that the substance would be present in po-
     tentially toxic levels in breast milk, and/or
b    on the basis of results of one or two generation studies in animals which indicate the presence of ad-
     verse effects on the offspring due to transfer in the milk, and/or
c    on the basis of evidence in humans indicating a risk to babies during the lactational period.
     Substances which are known to accumulate in the body and which subsequently may be released in-
     to milk during lactation may be labelled with R33 and R64.
Directive (93/21/EEC) of the European Community                                                             33
</pre>

====================================================================== Einde pagina 33 =================================================================

<br><br>====================================================================== Pagina 34 ======================================================================

<pre>Annex        D
             Fertility and developmental toxicity
             studies
Table 1 Fertility studies with chlorofom.
authors       species        route       experimen-   dose          findings                                    remarks
                                         tal period
Gulati et al. CD-1 mice      oral (gava- continuous   8, 20, 50     Body weight at the high dose chloroform Only abstract
(1997)                       ge)         breeding     mg/kg/bwb     treatment had no apparent effect on either of the study
                                         protocola.                 parental (F sub 0) or F sub 1 generation. F available.
                                                                    sub 1 generation males in the high dose
                                                                    group showed significantly increased epi-
                                                                    didymal weights. However, the epididy-
                                                                    mal sperm motility, sperm count and
                                                                    sperm morphology were not affected. F
                                                                    sub 1 females in the high dose group sho-
                                                                    wed increased liver weight and there were
                                                                    signs of hepatocellular degeneration.
a
    Both male and female mice (20 pairs per treatment group, 40 pairs for control animals) were dosed daily for 7 days prior
    to and during a 98-day cohabitation period. The F sub 1 generation from the control and high dose groups were also
    evaluated.
b
    It was estimated that the actual doses received were 6.6, 16, and 41 mg/kg bw in the low, mid and high dose groups, res-
    pectively.
             Fertility and developmental toxicity studies                                                                    34
</pre>

====================================================================== Einde pagina 34 =================================================================

<br><br>====================================================================== Pagina 35 ======================================================================

<pre>Table 2.1 Developmental toxicity studies with chlorofor.
authors        species     route      experimen- dose        findings                                         remarks
                                      tal period
Schwetz        Sprague-    inhalation days 6-15  0, 147, 490 At all dose levels statistically significant de- At 1470 mg/m3 the
et al. (1974) Dawley                  of gesta-  and 1470    crease in maternal weight gain and food con-     high level of early
               rat                    tion for 7 mg/m3       sumption.                                        embryonic death
                                      h/day                  At 147 mg/m3 retarded foetal development.        may mask malfor-
                                                             At 490 mg/m3 a significant number of litters     mations.
                                                             containing pups (number unspecified) with
                                                             missing or shortened tail and imperforate
                                                             anus, retarded foetal development.
                                                             At 1470 mg/m3 the number of litters available
                                                             for analysis was severely reduced because of a
                                                             decrease in the rate of conception and a high
                                                             incidence of foetal resorption. Terata were not
                                                             observed in this group.
Thompson       Sprague-    oral       days 6-15  0, 20, 50,  At 50 and 126 mg/kg bw decreased body
et al. (1974) Dawley                  of gesta-  126 mg/kg   weight gain. At 126 mg/kg bw decreased food
               rat                    tion       bw/day      consumption, maternal kidney and liver da-
                                                             mage, and statistically significantly decreased
                                                             foetal weight.
                                                             No teratogenic effects.
Thompson       rabbit      oral       days 6-18  0, 20, 35,  At 50 mg/kg bw decreased body weight gain,
et al. (1974)                         of gesta-  50 mg/kg    maternal kidney and liver damage, and statis-
                                      tion       bw/day      tically significantly decreased foetal weight.
                                                             No teratogenic effects.
Dilley         rat (speci- inhalation days 7-14  20.1 g/m3   Increased foetal mortality and decreased foe-    Study was poorly
et al. (1977) es ?)                   of gesta-              tal weight. No teratogenic effects.              reported.
                                      tion for ?
                                      h/day
              Fertility and developmental toxicity studies                                                                      35
</pre>

====================================================================== Einde pagina 35 =================================================================

<br><br>====================================================================== Pagina 36 ======================================================================

<pre>Table 2.2 Developmental toxicity studies with chloroform.
authors       species    route      experimen- dose           findings                                         remarks
                                    tal period
Murray et al. CF-1       inhalation days 1-7, 0, 147          Maternal toxicity consisted of a decrease in     Cleft palates were
(1979)        mouse                 6-15, 8-15 mg/m3          body weight gain. Also slightly less food and    seen predominant-
                                    of gesta-                 water consumption was observed.                  ly in foetuses with
                                    tion 7                    In all groups decreased ossification (not signi- retarded growth.
                                    h/day                     ficant)
                                                              Exposure from day 8 to 15 resulted in a signi-
                                                              ficant elevation in the incidence of cleft pala-
                                                              te, and significant decreased foetal weight
                                                              and rump-length.
                                                              Exposure from day 1 to 7 reduced litter size
                                                              (increase of resorptions) but no malforma-
                                                              tions. Decreased mean foetal body weight and
                                                              crown-rump length were also observed from
                                                              day 8 to 15.
Ruddick et    Sprague-   oral       days 6-15    0, 100, 200, Maternal weight gain was depressed in all        The observed foe-
al. (1983)    Dawley                of gesta-    400 mg/kg groups receiving Chloroform. Chloroform ad-         totoxicity was con-
              rat                   tion         bw/day       ministration caused decreased maternal hae-      sidered to be the
                                                              moglobin and haematocrit values at all dose      result of the mater-
                                                              levels and also produced increased serum in-     nal toxicity (WGD
                                                              organic phosphorus and cholesterol at the hig-   1987). Only ab-
                                                              hest dose. Liver enlargement was observed at     stract available.
                                                              all dose levels of Chloroform. Evidence of a
                                                              foetotoxic response (not specified) was obser-
                                                              ved with Chloroform, but teratogenic effects
                                                              were not observed.
Baeder and    Wistar     inhalation days 7-16    0, 30, 100,  30 ppm: 2 dams with only resorptions (vs 0 in
Hoffman       rats                  of gesta-    300 ppm      control); decreased foetal length
(1988)                              tion for                  100 ppm: gestation day 7-17 decreased body
                                    7h/day                    weight and food intake, 3 dams with only re-
                                                              sorptions, decreased foetal length.
                                                              300 ppm: gestation day 7-17 decreased body
                                                              weight and food intake; 8 dams with only re-
                                                              sorptions; decreased foetal length and weight.
             Fertility and developmental toxicity studies                                                                        36
</pre>

====================================================================== Einde pagina 36 =================================================================

<br><br>====================================================================== Pagina 37 ======================================================================

<pre>Table 2.3 Developmental toxicity studies with chloroform
authors      species    route       experimen- dose           findings                                               remarks
                                    tal period
Baeder and   Wistar     inhalation days 7-16     0, 3, 10, 30 0 ppm: no dams with resorptions, 3.2% small foetuses
Hoffman      rats                   of gesta-    ppm          3 ppm: 0 dams with only resorptions; 14.2% small foe-
(1991)                              tion for                  tuses (not statistically significant)
                                    7h/day                    10 ppm: 0 dams with only resorptions, 24% small foetu-
                                                              ses.
                                                              30 ppm: slightly reduced food intake; 1 dams with only
                                                              resorptions; 26.9% small foetuses.
           Fertility and developmental toxicity studies                                                                    37
</pre>

====================================================================== Einde pagina 37 =================================================================

<br><br>====================================================================== Pagina 38 ======================================================================

<pre>Annex E
      Abbreviations
                 Abbreviations used:
                            body weight
                            day
                            female(s)
                            intraperitoneal
                            intravenous
                            male(s)
                            number
                            no adverse effect level
                            Organisation for Economic Cooperation and Development
                            postnatal
      Abbreviations                                                               38
</pre>

====================================================================== Einde pagina 38 =================================================================

<br><br>