S.I. No. 116/2003 - European Communities (Classification, Packaging, Labelling and Notification of Dangerous Substances) Regulations 2003

• (b) or when experimental data are available, according to the criteria described under Sections 3.1.2 excluding the carcinogenic, mutagenic and toxic to reproduction properties referred to under 3.1.2 (e) which must be evaluated by a conventional method referred to in Article 6 and Annex II, Parts A 7 - 9 and B 6 of Directive 1999/45/EC.

Note: Without prejudice to requirements of Directive 91/414/EEC, only where it can be scientifically demonstrated by the person responsible for placing the preparation on the market that the toxicological properties of the preparation cannot correctly be determined by the method outlined in paragraph 3.1.3 (a), or on the basis of existing test results on animals, the methods outlined in paragraph 3.1.3 (b) may be used, provided they are justified or specifically authorised under Article 12 of Directive 86/609/EEC.

Whichever method is used for the evaluation of the danger of a preparation, all the dangerous effects on health as defined in Annex II, Part B of Directive 1999/45/EC must be taken into consideration.

3.1.4. When the classification is to be established from experimental results obtained in animal tests the results should have validity for man in that the tests reflect, in an appropriate way, the risks to man.

3.1.5. The acute oral toxicity of substances or preparations placed on the market may be established either by a method permitting assessment of the LD50 value, or by determining the discriminating dose (the fixed dose method), or by determining the range of exposure where lethality is expected (the acute toxic class method).

3.1.5.1. The discriminating dose is the dose which causes evident toxicity but not mortality and must be one of the four dosage levels specified in Annex V (5, 50, 500 or 2 000 mg per kg body weight).

The concept ‘evident toxicity’ is used to designate toxic effects, after exposure to the substance tested, which are so severe that exposure to the next highest fixed dose would probably lead to mortality.

The results of testing at a particular dose following the fixed dose method may be either:

• less than 100 % survival,

• 100 % survival, but evident toxicity,

• 100 % survival, but no evident toxicity.

In the criteria in sections 3.2.1, 3.2.2 and 3.2.3 only the final test result is shown. The 2 000 mg/kg dose should be used primarily to obtain information on the toxic effects of substances which are of low acute toxicity and which are not classified on the basis of acute toxicity.

The fixed dose method requires in some cases testing at higher or lower doses, if not already tested at the relevant dose level. Refer also to the evaluation table in test method B.1 bis.

3.1.5.1. The range of exposure where lethality is expected is derived from the observed absence or presence of substance related mortality following the acute toxic class method. For initial testing one of three fixed starting doses (25, 200 or 2 000 mg per kg body weight) is used.

The acute toxic class method requires in some cases testing at higher or lower doses, if not already tested at the relevant dose level. Refer also to the test procedure flow charts in test method B.1 tris of Annex V.

3.2. Criteria for classification, choice of symbols, indication of danger, choice of risk phrases

3.2.1. Very toxic

Substances and preparations shall be classified as very toxic, and assigned the symbol ‘T+’ and indication of danger ‘very toxic’ in accordance with the criteria specified below.

Risk phrases shall be assigned in accordance with the following criteria:

R28 Very toxic if swallowed

Acute toxicity results:

• LD50 oral, rat ≤ 25 mg/kg,

• less than 100 % survival at 5 mg/kg oral, rat by the fixed dose procedure, or

• high mortality at doses ≤ 25 mg/kg oral, rat, by the acute toxic class method (for test result interpretation see flow charts in Annex 2 of test method B.1 tris of Annex V).

R27 Very toxic in contact with skin

Acute toxicity results:

• LD50 dermal, rat or rabbit: ≤ 50 mg/kg.

R26 Very toxic by inhalation

Acute toxicity results:

• LC50 inhalation, rat, for aerosols or particulates: ≤ 0.25 mg/litre/4hr,

• LC50 inhalation, rat, for gases and vapours: ≤ 0.5 mg/litre/4hr.

R39 Danger of very serious irreversible effects

• Strong evidence that irreversible damage other than the effects referred to in Chapter 4 is likely to be caused by a single exposure by an appropriate route, generally in the above-mentioned dose range.

In order to indicate the route of administration/exposure one of the following combinations shall be used: R39/26, R39/27, R39/28, R39/26/27, R39/26/28, R39/27/28, R39/26/27/28.

3.2.2. Toxic

Substances and preparations shall be classified as toxic and assigned the symbol ‘T’ and the indication of danger ‘toxic’ in accordance with the criteria specified below. Risk phrases shall be assigned in accordance with the following criteria.

R25 Toxic if swallowed

Acute toxicity results:

• LD50 oral, rat: 25 < LD50 ≤ 200 mg/kg,

• Discriminating dose, oral, rat, 5 mg/kg: 100 % survival but evident toxicity, or

• high mortality in the dose range > 25 to ≤ 200 mg/kg oral, rat, by the acute toxic class method (for test result interpretation see flow charts in Annex 2 of test method B.1 tris of Annex V).

R24 Toxic in contact with skin

Acute toxicity results:

• LD50 dermal, rat or rabbit: 50 < LD50 ≤ 400 mg/kg.

R23 Toxic by inhalation

Acute toxicity results:

• LC50 inhalation, rat, for aerosols or particulates: 0.25 < LC50 ≤ 1 mg/litre/4hr,

• LC50 inhalation, rat, for gases and vapours: 0.5 < LC50 ≤ 2 mg/litre/4hr.

R39 Danger of very serious irreversible effects

• strong evidence that irreversible damage other than the effects referred to in Chapter 4 is likely to be caused by a single exposure by an appropriate route, generally in the above-mentioned dose range.

In order to indicate the route of administration/exposure one of the following combinations shall be used: R39/23, R39/24, R39/25, R39/23/24, R39/23/25, R39/24/25, R39/23/24/25.

R48 Danger of serious damage to health by prolonged exposure

• serious damage (clear functional disturbance or morphological change which have toxicological significance) is likely to be caused by repeated or prolonged exposure by an appropriate route.

Substances and preparations are classified at least as toxic when these effects are observed at levels of one order of magnitude lower (i.e. 10-fold) than those set out for R48 in Section 3.2.3.

In order to indicate the route of administration/exposure one of the following combinations shall be used: R48/23, R48/24, R48/25, R48/23/24, R48/23/25, R48/24/25, R48/23/24/25.

3.2.3. Harmful

Substances and preparations shall be classified as harmful and assigned the symbol ‘Xn’ and the indication of danger ‘harmful’ in accordance with the criteria specified below. Risk phrases shall be assigned in accordance with the following criteria:

R22 Harmful if swallowed

Acute toxicity results:

• LD50 per oral, rat: 200 < LD50 ≤ 2000 mg/kg,

• discriminating dose, oral, rat, 50 mg/kg: 100 % survival but evident toxicity,

• less than 100 % survival at 500 mg/kg, rat oral by the fixed dose procedure. Refer to the evaluation table in the test method B.1 bis of Annex V, or

• high mortality in the dose range > 200 to ≤ 2 000 mg/kg oral, rat, by the acute toxic class method (for test result interpretation see flow charts in Annex 2 of test method B.1 tris of Annex V).

R21 Harmful in contact with skin

Acute toxicity results:

• LD50 dermal, rat or rabbit: 400 < LD50 ≤ 2 000 mg/kg.

R20 Harmful by inhalation

Acute toxicity results:

• LC50 inhalation, rat, for aerosols or particulates: 1 < LC50 ≤ 5 mg/litre/4hr,

• LC50 inhalation, rat, for gases or vapours: 2 < LC50 ≤ 20 mg/litre/4hr.

R65 Harmful: may cause lung damage if swallowed

Liquid substances and preparations presenting an aspiration hazard in humans because of their low viscosity:

• (a) For substances and preparations containing aliphatic, alicyclic and aromatic hydrocarbons in a total concentration equal to or greater than 10 % and having either

• a flow time of less than 30 sec. in a 3 mm ISO cup according to ISO 2431 (April 1996 / July 1999 edition) relating to ‘Paints and varnishes - Determination of flow time by use of flow cups’,

• a kinematic viscosity measured by a calibrated glass capillary viscometer in accordance with ISO 3104/3105 of less than 7 × 10^-6 m²/sec at 40° C (ISO 3104 1994 edition, relating to ‘Petroleum products - Transparent and opaque liquids - Determination of kinematic viscosity and calculation of dynamic viscosity’; ISO 3105, 1994 edition, relating to ‘Glass capillary kinematic viscometers - Specifications and operating instructions’), or

• a kinematic viscosity derived from measurements of rotational viscometry in accordance with ISO 3219 of less than 7 × 10^-6 m²/sec at 40° C (ISO 3219, 1993 edition, relating to ‘Plastics - Polymers/resins in the liquid state or as emulsions or dispersions - Determination of viscosity using a rotational viscometer with defined shear rate’).

Note that substances and preparations meeting these criteria need not be classified if they have a mean surface tension greater than 33mN/m at 25° C as measured by the du Nouy tensiometer or by the test methods shown in Annex V Part A.5.

• (b) For substances and preparations, based on practical experience in humans.

R68 Possible risk of irreversible effects

• strong evidence that irreversible damage other than the effects referred to in Chapter 4 is likely to be caused by a single exposure by an appropriate route, generally in the above-mentioned dose range.

In order to indicate route of administration/exposure one of the following combinations shall be used: R68/20, R68/21, R68/22, R68/20/21, R68/20/22, R68/21/22, R68/20/21/22.

R48 Danger of serious damage to health by prolonged exposure

• serious damage (clear functional disturbance or morphological change which has toxicological significance) is likely to be caused by repeated or prolonged exposure by an appropriate route.

Substances and preparations are classified at least as harmful when these effects are observed at levels of the order of:

• oral, rat ≤ 50 mg/kg (bodyweight)/day,

• dermal, rat or rabbit ≤ 100 mg/kg (bodyweight)/day,

• inhalation, rat ≤ 0.25 mg/l, 6h/day.

These guide values can apply directly when severe lesions have been observed in a sub-chronic (90 days) toxicity test. When interpreting the results of a sub-acute (28 days) toxicity test these figures should be increased approximately three fold. If a chronic (two years toxicity test is available it should be evaluated on a case-by-case basis. If results of studies of more than one duration are available, then those from the study of the longest duration should normally be used.

In order to indicate route of administration/exposure one of the following combinations shall be used: R48/20, R48/21, R48/22, R48/20/21, R48/20/22, R48/21/22, R48/20/21/22.

3.2.3.1. Comments regarding volatile substances

For certain substances with a high-saturated vapour concentration evidence may be available to indicate effects that give cause for concern. Such substances may not be classified under the criteria for health effects in this guide (3.2.3) or not covered by Section 3.2.8. However, where there is appropriate evidence that such substances may present a risk in normal handling and use then classification on a case-by-case basis in Annex I may be necessary.

3.2.4. Comments regarding the use of R48

Use of this risk phrase refers to the specific range of biological effects within the terms described below. For application of this risk phrase serious damage to health is to be considered to include death, clear functional disturbance or morphological changes which are toxicologically significant. It is particularly important when these changes are irreversible. It is also important to consider not only specific severe changes in a single organ or biological system but also generalised changes of a less severe nature involving several organs, or severe changes in general health status.

When assessing whether there is evidence for these types of effects reference should be made to the following guidelines:

• (a) substance-related deaths;

• (b) (i) major functional changes in the central or peripheral nervous systems including sight, hearing and the sense of smell, assessed by clinical observations or other appropriate methods (e.g. electrophysiology);

• (ii) major functional changes in other organ systems (for example the lung);

• (c) any consistent changes in clinical biochemistry, haematology or urinalysis parameters which indicate severe organ dysfunction. Haematological disturbances are considered to be particularly important if the evidence suggests that they are due to decreased bone marrow production of blood cells;

• (d) severe organ damage noted on microscopic examination following autopsy:

• (i) widespread or severe necrosis, fibrosis or granuloma formation in vital organs with regenerative capacity (e.g. liver);

• (ii) severe morphological changes that are potentially reversible but are clear evidence of marked organ dysfunction (e.g. severe fatty change in the liver severe acute tubular nephrosis in the kidney, ulcerative gastritis); or

• (iii) evidence of appreciable cell death in vital organs incapable of regeneration (e.g. fibrosis of the myocardium or dying back of a nerve) or in stem cell populations (e.g. aplasia or hypoplasia of the bone marrow).

The above evidence will most usually be obtained from animal experiments. When considering data derived from practical experience special attention should be given to exposure levels.

The use of this risk phrase is restricted to ‘serious damage to health by prolonged exposure’. A number of substance-related effects may be observed in both humans and animals that would not justify the use of R48. These effects are relevant when attempting to determine a no-effect level for a chemical substance. Examples of well documented changes which would not normally justify classification with R48, irrespective of their statistical significance, include:

• (a) clinical observations or changes in bodyweight gain, food consumption or water intake, which may have some toxicological importance but which do not, by themselves, indicate ‘serious damage’;

• (b) small changes in clinical biochemistry, haematology or urinalysis parameters which are of doubtful or minimal toxicological importance;

• (c) changes in organ weights with no evidence of organ dysfunction;

• (d) adaptative responses (e.g. macrophage migration in the lung, liver hypertrophy and enzyme induction, hyperplastic responses to irritants). Local effects on the skin produced by repeated dermal application of a substance which are more appropriately classified with R38 ‘irritating to skin’; or

• (e) where a species-specific mechanism of toxicity (e.g. specific metabolic pathways) has been demonstrated.

3.2.5. Corrosive

The substance or preparation shall be classified as corrosive and assigned the symbol ‘C’ and the indication of danger ‘corrosive’ in accordance with the following criteria:

• A substance or a preparation is considered to be corrosive if, when it is applied to healthy intact animal skin, it produces full thickness destruction of skin tissue on at least one animal during the test for skin irritation cited in Annex V or during an equivalent method.

• Classification can be based on the results of a validated in vitro test, such as that cited in Annex V (B.40. Skin corrosion: rat skin transcutaneous electrical resistance assay and human skin model assay.)

• A substance or a preparation should also be considered corrosive if the result can be predicted, for example from strongly acid or alkaline reactions indicated by a pH of 2 or less or 11.5 or greater. However, where extreme pH is the basis for classification acid/alkali reserve⁶⁹ may also be taken into consideration. If consideration of alkali/acid reserve suggests the substance or preparation may not be corrosive then further testing should be carried out to confirm this, preferably by use of an appropriate validated in vitro test. Consideration of acid/alkali reserve should not be used alone to exonerate substances or preparations from classification as corrosive.

Risk phrases shall be assigned in accordance with the following criteria:

R35 Causes severe burns

• if, when applied to healthy intact animal skin, full thickness destruction of skin tissue occurs as a result of up to three minutes exposure, or if this result can be predicted.

R34 Causes burns

• if, when applied to healthy intact animal skin, full thickness destruction of skin tissue occurs as a result of up to four hours exposure, or if this result can be predicted,

• organic hydroperoxides, except where evidence to the contrary is available.

Notes:

Where classification is based on results of a validated in vitro test R35 or R34 should be applied according to the capacity of the test method to discriminate between these.

Where classification is based upon consideration of extreme pH alone, R35 should be applied.

3.2.6. Irritant

Substances and preparations shall be classified as irritant and assigned the symbol ‘Xi’ and the indication of danger ‘irritant’ in accordance with the criteria given below.

3.2.6.1. Inflammation of the skin

The following risk phrase shall be assigned in accordance with the criteria given:

R38 Irritating to skin

• Substances and preparations which cause significant inflammation of the skin which persists for at least 24 hours after an exposure period of up to four hours determined on the rabbit according to the cutaneous irritation test method cited in Annex V.

Inflammation of the skin is significant if:

• (a) the mean value of the scores for either erythema and eschar formation or oedema formation, calculated over all the animals tested, is 2 or more; or

• (b) in the case where the Annex V test has been completed using three animals either erythema and eschar formation or oedema formation equivalent to a mean value of 2 or more calculated for each animal separately has been observed in two or more animals.

In both cases all scores at each of the reading times (24, 48 and 72 hr) for an effect should be used in calculating respective mean values.

Inflammation of the skin is also significant if it persists in at least two animals at the end of the observation time. Particular effects e.g. hyperplasia, scaling, discoloration, fissures, scabs and alopecia should be taken into account.

Relevant data may also be available from non-acute animal studies (see comments on R48 section 2.d). These are considered significant if the effects seen are comparable to those described above.

• Substances and preparations which cause significant inflammation of the skin, based on practical observations in humans on immediate, prolonged or repeated contact.

• Organic peroxides, except where evidence to the contrary is available.

Paresthesia:

Paresthesia caused in humans by skin contact with pyrethroid pesticides is not regarded as an irritant effect justifying classification as Xi; R38. The S-phrase S24 should however be applied for substances seen to cause this effect.

3.2.6.2. Ocular lesions

The following risk phrases shall also be assigned in accordance with the criteria given:

R36 Irritating to eyes

• Substances and preparations which, when applied to the eye of the animal, cause significant ocular lesions which occur within 72 hours after exposure and which persist for at least 24 hours.

Ocular lesions are significant if the mean scores of the eye irritation test cited in Annex V have any of the following values:

• cornea opacity equal to or greater than 2 but less than 3,

• iris lesion equal to or greater than 1 but not greater than 1.5,

• redness of the conjunctivae equal to or greater than 2.5,

• oedema of the conjunctivae (chemosis) equal to or greater than 2,

or, in the case where the Annex V test has been completed using three animals if the lesions on two or more animals, are equivalent to any of the above values except that for iris lesion the value should be equal to or greater than 1 but less than 2 and for redness of the conjunctivae the value should be equal to or greater than 2.5.

In both cases all scores at each of the reading times (24, 48 and 72 hours) for an effect should be used in calculating the respective mean values.

• Substances or preparations which cause significant ocular lesions, based on practical experience in humans.

• Organic peroxides except where evidence to the contrary is available.

R41 Risk of serious damage to eyes

• Substances and preparations which, when applied to the eye of the animal cause severe ocular lesions which occur within 72 hours after exposure and which persist for at least 24 hours.

Ocular lesions are severe if the means of the scores of the eye irritation test in Annex V have any of the values:

• cornea opacity equal to or greater than 3,

• iris lesion greater than 1.5.

The same shall be the case where the test has been completed using three animals if these lesions on two or more animals, have any of the values:

• cornea opacity equal to or greater than 3,

• iris lesion equal to 2.

In both cases all scores at each of the reading times (24, 48 and 72 hours) for an effect should be used in calculating the respective mean values.

Ocular lesions are also severe when they are still present at the end of the observation time.

Ocular lesions are also severe if the substance or preparation causes irreversible colouration of the eyes.

• Substances and preparations which cause severe ocular lesions, based on practical experience in humans.

Note:

When a substance or preparation is classified as corrosive and assigned R34 or R35, the risk of severe damage to eyes is considered implicit and R41 is not included in the label.

3.2.6.3. Respiratory system irritation

The following risk phrase shall be assigned in accordance with the criteria given:

R37 Irritating to respiratory system

Substances and preparations which cause serious irritation to the respiratory system based on:

• practical observation in humans

• positive results from appropriate animal tests.

Comments regarding the use of R37

In interpreting practical observations in humans, care should be taken to distinguish between effects which lead to classification with R48 (see section 3.2.4.) from those leading to classification with R37. Conditions normally leading to classification with R37 are reversible and usually limited to the upper airways.

Positive results from appropriate animal tests may include data obtained in a general toxicity test, including histopathological data from the respiratory system. Data from the measurement of experimental bradypnea may also be used to assess airway irritation.

3.2.7. Sensitisation

3.2.7.1. Sensitisation by inhalation

Substances and preparations shall be classified as sensitising and assigned the symbol ‘Xn’, the indication of danger ‘Harmful’ and the risk phrase R42 in accordance with the criteria given below.

R42 May cause sensitisation by inhalation

• if there is evidence that the substance or preparation can induce specific respiratory hypersensitivity;

• where there are positive results from appropriate animal tests; or

• if the substance is an isocyanate, unless there is evidence that the specific isocyanate does not cause respiratory hypersensitivity

Comments regarding the use of R42

Human evidence

Evidence that the substance or preparation can induce specific respiratory hypersensitivity will normally be based on human experience. In this context hypersensitivity is normally seen as asthma, but other hypersensitivity reactions such as rhinitis and alveolitis are also considered. The condition will have the clinical character of an allergic reaction. However, immunological mechanisms do not have to be demonstrated.

When considering the evidence from human exposure, it is necessary for a decision on classification to take into account in addition to the evidence from the cases:

• the size of the population exposed

• the extent of exposure.

The evidence referred to above could be:

• clinical history and data from appropriate lung function tests related to exposure to the substance, confirmed by other supportive evidence which may include:

• a chemical structure related to substances known to cause respiratory hypersensitivity;

• an in vivo immunological test (e.g. skin prick test);

• an in vitro immunological test (e.g. serological analysis);

• studies indicating other specific but non-immunological mechanisms of action, e.g. repeated low-level irritation, pharmacologically mediated effects; or

• data from a positive bronchial challenge test with the substance conducted according to accepted guidelines for the determination of a specific hypersensitivity reaction.

Clinical history should include both medical and occupational history to determine a relationship between exposure to a specific substance or preparation and development of respiratory hypersensitivity. Relevant information includes aggravating factors both in the home and workplace, the onset and progress of the disease, family history and medical history of the patient in question. The medical history should also include a note of other allergic or airway disorders from childhood, and smoking history.

The results of positive bronchial challenge tests are considered to provide sufficient evidence for classification on their own. It is however recognised that in practice many of the examinations listed above will already have been carried out.

Substances that elicit symptoms of asthma by irritation only in people with bronchial hyperreactivity should not be assigned R42.

Animal studies

Data from tests which may be indicative of the potential of a substance or preparation to cause sensitisation by inhalation in humans may include:

• IgE measurements (e.g. in mice), or

• specific pulmonary responses in guinea pigs.

3.2.7.2. Sensitisation by skin contact

Substances and preparations shall be classified as sensitising and assigned the symbol ‘Xi’, the indication of danger ‘Irritant’ and the risk phrase R43 in accordance with the criteria given below:

R43 May cause sensitisation by skin contact

• If practical experience shows the substance or preparation to be capable of inducing a sensitisation by skin contact in a substantial number of persons, or

• where there are positive results from an appropriate animal test.

Comments regarding the use of R43

Human evidence

The following evidence (practical experience) is sufficient to classify a substance or preparation with R43:

• Positive data from appropriate patch testing, normally in more than one dermatological clinic, or

• Epidemiological studies showing allergic contacts dermatitis caused by the substance or preparation. Situations in which a high proportion of those exposed exhibit characteristic symptoms are to be looked at with special concern, even if the number of cases is small, or

• Positive data from experimental studies in man (see also 3.1.1).

The following is sufficient to classify a substance with R43 when there is supportive evidence:

• Isolated episodes of allergic contact dermatitis, or

• Epidemiological studies where chance, bias or confounders have not been ruled out fully with reasonable confidence.

Supportive evidence may include:

• data from animal tests performed according to existing guidelines, with a result that does not meet the criteria given in the section on animal studies but is sufficiently close to the limit to be considered significant, or

• data from non-standard methods, or

• appropriate structure-activity relationships.

Animal studies

Positive results from appropriate animal tests are:

• in the case of the adjuvant type test method for skin sensitisation detailed in Annex V or in the case of other adjuvant-type test methods, a response of at least 30% of the animals is considered positive;

• for any other test method a response of at least 15% of the animals is considered positive.

3.2.7.3. Immunological contact urticaria

Some substances or preparations, which meet the criteria for R42 may in addition cause immunological contact urticaria. In these cases, information concerning contact urticaria should be included by the use of appropriate S-phrases, usually S24 and S36/37, and in the Safety Data Sheet.

For substances or preparations, which produce signs of immunological contact urticaria which do not fulfill the criteria for R42, consideration should be given to classification with R43.

There is no recognised animal model available to identify substances which cause immunological contact urticaria. Therefore, classification will normally be based on human evidence which will be similar to that for skin sensitisation (R43).

3.2.8. Other toxicological properties

Additional risk phrases shall be assigned in accordance with the following criteria (based on experience obtained during compilation of Annex I) to substances and preparations classified by virtue of 2.2.1. to 3.2.7. above and/or chapters 4 and 5:

R29 Contact with water liberates toxic gas

For substances and preparations which in contact with water or damp air, evolve very toxic/toxic gases in potentially dangerous amounts, e.g. aluminium phosphide phosphorus pentasulphide.

R29 Contact with acids liberates toxic gas

For substances and preparations which react with acids to evolve toxic gases in dangerous amounts, e.g. sodium hypochlorite, barium polysulphide. For substances used by members of the general public, the use of S50 (do not mix with … (to be specified by the manufacturer)) would be more suitable.

R32 Contact with acids liberates very toxic gas

For substances and preparations which react with acids to evolve very toxic gases in dangerous amounts; e.g. salts of hydrogen cyanide, sodium azide. For substances used by members of the general public, the use of S50 (do not mix with … (to be specified by the manufacturer)) would be more suitable.

R33 Danger of cumulative effects

For substances and preparations when accumulation in the human body is likely and may cause some concern which, however, is not sufficient to justify the use of R48.

For comments on the use of this R-phrase see Section 4.2.3.3 for substances and Annex V, Part A.3. of Directive 1999/45/EC for preparations.

R64 May cause harm to breastfed babies

For substances and preparations which are absorbed by women 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.

For comments on the use of this R-phrase see Section 4.2.3.3 for substances and Annex V, Part A.4. of Directive 1999/45/EC for preparations.

R66 Repeated exposure may cause skin dryness or cracking

For substances and preparations which may cause concern as a result of skin dryness flaking or cracking but which do not meet the criteria for R38 based on either:

• practical observation after normal handling and use, or

• relevant evidence concerning their predicted effects on the skin.

See also paragraphs 1.6 and 1.7.

R67 Vapours may cause drowsiness and dizziness

For volatile substances and preparations containing such substances which cause clear symptoms of central nervous system depression by inhalation and which are not already classified with respect to acute inhalation toxicity (R20, R23, R26, R68/20, R39/23 or R39/26).

The following evidence may be used:

• (a) Data from animal studies showing clear signs of CNS depression such as narcotic effects, lethargy, lack of co-ordination (including loss of righting reflex) and ataxia either:

• at concentrations/exposure times not exceeding 20 mg/1/4h or,

• for which the ratio of the effect concentration at ≤ 4 h to the saturated vapour concentration (SVC) at 20°C is ≤ 1/10

• (b) Practical experience in humans (e.g. narcosis, drowsiness, reduced alertness, loss of reflexes, lack of co-ordination, vertigo) from well documented reports under comparable exposure conditions to the effects specified above for animals.

See also Paragraphs 1.6 and 1.7.

For other supplementary risk phrases see Section 2.2.6.

4.1. Introduction

4.1.1. This Chapter sets out the procedure for the classification of substances which may have the effects mentioned below. For preparations see Section 4.2.4.

4.1.2. If a manufacturer, distributor or importer has information available which indicates that a substance should be classified and labelled in accordance with the criteria given in Section 4.2.1, 4.2.2 or 4.2.3, he shall provisionally label the substance in accordance with these criteria, on the basis of the assessment of the evidence by a competent person.

4.1.3. The manufacturer, distributor or importer shall submit as soon as possible a document summarising all relevant information to one Member State in which the substance is placed on the market. Relevant information in this context comprises in particular all available published and unpublished information required for appropriate classification of the substance in question, on the basis of the intrinsic properties according to the categories laid down in Article 2 (2) and in accordance with the criteria in this Annex. The submitted summary document should include a bibliography containing all relevant references, including any relevant unpublished data.

4.1.4. Furthermore, a manufacturer, distributor or importer who has new data which are relevant to the classification and labelling of a substance in accordance with the criteria given in Section 4.2.1., 4.2.2. or 4.2.3., shall submit this data as soon as possible to one Member State in which the substance is placed on the market.

4.1.5. To obtain as quickly as possible a harmonised classification for the Community by the procedure defined in Article 28 of Directive 67/548/EEC, Member States which have relevant information available justifying the classification of a substance in one of these categories whether submitted by the manufacturer or not, should forward such information together with suggestions for classification and labelling, to the Commission as soon as possible.

The Commission will forward to the other Member States the classification and labelling proposal that it receives. Any Member State may ask the Commission for the information it has received.

Any Member State which has good reason to believe that the suggested classification and labelling is inappropriate as far as the carcinogenic, mutagenic or reproductive toxicity effects are concerned shall notify the Commission thereof.

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.

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 assessment. 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:

Categories 1 and 2:

Substances classified as carcinogenic category 1 or 2 shall be assigned the symbol ‘T’ and the risk phrase

R45 May cause cancer

However, 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), shall be assigned the symbol ‘T’ and the risk phrase

R49 May cause cancer by inhalation

Category 3:

Substances classified as carcinogenic category 3 shall be assigned the symbol ‘Xn’ and the risk phrase

R40 Limited evidence of a carcinogenic effect

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 genotoxicity data, metabolic or biochemical studies, induction of benign tumours, structural relationship with other known carcinogens, or data from epidemiological studies suggesting an association.

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 date 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 arguments, 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 dose levels exceeding the ‘maximal tolerated dose’. The maximal tolerated dose is characterised 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 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 extrapolated 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 occurrence of neoplasms at sites and in strains where they are well known to occur spontaneously with a high incidence.

4.2.2 Mutagenic substances

4.2.2.1. For the purposes 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 mutagenic to man.

There is sufficient evidence to establish a causal association between human exposure to a substance and heritable genetic damage.

Category 2

Substances which should be regarded as if they are mutagenic to man.

There is sufficient evidence to provide a strong presumption that human exposure to the substance may result in the development of heritable genetic damage, generally on the basis of:

• appropriate animal studies,

• other relevant information.

Category 3

Substances which cause concern for man owing to possible mutagenic effects. There is evidence from appropriate mutagenicity studies, but this is insufficient to place the substance in Category 2.

4.2.2.2. The following symbols and specific risk phrases apply:

Categories 1 and 2:

Substances classified as mutagenic category 1 or 2 shall be assigned the symbol ‘T’ and the risk phrase

R46 May cause heritable genetic damage

Category 3:

Substances classified as mutagenic category 3 shall be assigned the symbol ‘Xn’ and the risk phrase

R68 Possible risk of irreversible effects

4.2.2.3. Comments regarding the categorisation of mutagenic substances

Definition of terms:

A mutation is permanent change in the amount or structure of the genetic material in an organism, resulting in a change of the phenotypic characteristics of the organism. The alterations may involve a single gene, a block of genes, or a whole chromosome. Effects involving single genes may be a consequence of effects on single DNA bases (point mutations) or of large changes, including deletions, within the gene. Effects on whole chromosomes may involve structural or numerical changes. A mutation in the germ cells in sexually reproducing organisms may be transmitted to the offspring. A mutagen is an agent that gives rise to an enhanced occurrence of mutations.

It should be noted that substances are classified as mutagens with specific reference to inherited genetic damage. However, the type of results leading to classification of chemicals in Category 3: ‘induction of genetically relevant events in somatic cells’, is generally also regarded as an alert for possible carcinogenic activity.

Method development for mutagenicity testing is an ongoing process. For many new tests no standardised protocols and evaluation criteria are presently available. For the evaluation of mutagenicity data the quality of the test performance and the degree of validation of the test method have to be considered.

Category 1

To place a substance in Category 1, positive evidence from human mutation epidemiology studies will be needed. Examples of such substances are not known to date. It is recognised that it is extremely difficult to obtain reliable information from studies on the incidence of mutations in human populations, or on possible increases in their frequencies.

Category 2

To place a substance in Category 2, positive results are needed from assays showing (a mutagenic effects, or (b) other cellular interactions relevant to mutagenicity, in germ cells of mammals in vivo, or (c) mutagenic effects in somatic cells of mammals in vivo in combination with clear evidence that the substance or a relevant metabolite reaches the germ cells.

With respect to placement in Category 2, at present the following methods are appropriate:

2 (a) in vivo germ cell mutagenicity assays:

• specific locus mutation test,

• heritable translocation test,

• dominant lethal mutation test.

These assays actually demonstrate the appearance of affected progeny or a defect in the developing embryo.

2 (b) in vivo assays showing relevant interaction with germ cells (usually DNA):

• assays for chromosomal abnormalities, as detected by cytogenetic analysis, including aneuploidy, caused by malsegregation of chromosomes,

• test for sister chromatid exchanges (SCEs),

• test for unscheduled DNA synthesis (UDS),

• assay of (covalent) binding of mutagen to germ cell DNA,

• assaying other kinds of DNA damage.

These assays provide evidence of a more or less indirect nature. Positive results in these assays would normally be supported by positive results from in vivo somatic cell mutagenicity assays, in mammals or in man (see under Category 3, preferably methods as under 3 (a)).

2 (c) in vivo assays showing mutagenic effects in somatic cells of mammals (see under 3 (a)) in combination with toxicokinetic methods, or other methodologies capable of demonstrating that the compound or a relevant metabolite reaches the germ cells.

For 2 (b) and 2 (c), positive results from host-mediated assays or the demonstration of unequivocal effects in in vitro assays can be considered as supporting evidence.

Category 3

To place a substance in Category 3, positive results are needed in assays showing (a mutagenic effects or (b) other cellular interaction relevant to mutagenicity, in somatic cells in mammals in vivo. The latter especially would normally be supported by positive results from in vitro mutagenicity assays.

For effects in somatic cells in vivo at present the following methods are appropriate:

3 (a) in vivo somatic cell mutagenicity assays:

• bone marrow micronucleus test or metaphase analysis,

• metaphase analysis of peripheral lymphocytes,

• mouse coat colour spot test.

3 (b) in vivo somatic cell DNA interaction assays:

• test for SCEs in somatic cells,

• test for UDS in somatic cells,

• assay for the (covalent) binding of mutagen to somatic cell DNA,

• assay for DNA damage, e.g. by alkaline elution, in somatic cells.

Substances showing positive results only in one or more in vitro mutagenicity assays should normally not be classified. Their further investigation using in vivo assays, however, is strongly indicated. In exceptional cases, e.g. for a substance showing pronounced responses in several in vitro assays, for which no relevant in vivo data are available, and which shows resemblance to known mutagens/carcinogens, classification in Category 3 could be considered.

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

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 as 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 results 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 occuring at around the same dose levels as other toxic effects, but which is 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.

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.

4.2.3.2. The following symbols and specific risk phrases apply:

Category 1

for substances that impair fertility in humans:

Substances classified as toxic to reproduction category 1 shall be assigned the symbol ‘T’ and the risk phrase

R60 May impair fertility

for substances that cause developmental toxicity:

Substances classified as toxic to reproduction category 1 shall be assigned the symbol ‘T’ and the risk phrase

R61 May cause harm to the unborn child

Category 2

for substances that should be regarded as if they impair fertility in humans:

Substances classified as toxic to reproduction category 2 shall be assigned the symbol ‘T’ and the risk phrase

R60 May impair fertility

for substances that should be regarded as if they cause developmental toxicity in humans:

Substances classified as toxic to reproduction category 2 shall be assigned the symbol ‘T’ and the risk phrase

R61 May cause harm to the unborn child.

Category 3

for substances which cause concern for human fertility:

Substances classified as toxic to reproduction category 3 shall be assigned the symbol ‘Xn’ and the risk phrase

R62 Possible risk of impaired fertility

for substances which cause concern for humans owing to possible developmental toxic effects:

Substances classified as toxic to reproduction category 3 shall be assigned the symbol ‘Xn’ and the risk phrase

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 headings 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 interfere 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 development, 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 (teratogenic effects), functional defects, peri-postnatal defects, and impaired postnatal mental or physical 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 animal 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 administered, 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 similar reasons it may be that classification in Category 3, or even no classification, will be warranted.

Annex V to Directive 67/548/EEC 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 levels 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 necessarily 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 anti-fertility agents or other information from humans which would 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 appropriate.

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 evidence 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 studies 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 associated 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 results of such studies must be interpreted with caution and on their own would not normally lead to classification.

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 possibility that the effects may have been due to non-specific influences such as generalised toxicity cannot be excluded.

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.).

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 reproduction’, 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 potentially 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 adverse 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 into milk during lactation may be labelled with R33 and R64.

4.2.4. Procedure for the classification of preparations concerning specific effects on health

If a preparation contains one or more substances classified with respect to the criteria laid out above, it must be classified according to the criteria referred to in Annex II, Part A. 7. — 9. and Part B. 6. of Directive 1999/45/EC (the concentration limits are either in Annex I of this Directive, or in Annex II, Part B. 6. of Directive 1999/45/EC where the substance or substances under consideration do not appear in Annex I or appear in it without concentration limits).

5.1. Introduction

The primary objective of classifying substances and preparations dangerous for the environment is to alert the user to the hazards these substances and preparations present to ecosystems. Although the present criteria refer to aquatic ecosystems it is recognised that certain substances and preparations may simultaneously or alternatively affect other ecosystems whose constituents may range from soil microflora and microfauna to primates.

The criteria set out below follow directly from the test methods set out in Annex V in so far as they are mentioned. The test methods required for the ‘base set’ referred to in Annex VII are limited and the information derived from them may be insufficient for an appropriate classification. Classification may require additional data derived from Level 1 (Annex VIII) or other equivalent studies. Furthermore, classified substances may be subject to review in the light of other new data.

For the purposes of classification and labelling and having regard to the current state of knowledge such substances and preparations are divided into two groups according to their acute and/or long-term effects in aquatic systems or their acute and/or long-term effects in non-aquatic systems.

5.1.1. The classification of substances is usually made on the basis of experimental data for acute aquatic toxicity, degradation, and log Pow (or BCF if available).

5.1.2. The classification of preparations shall normally be carried out on the basis of a conventional method referred to in Article 7 and Annex III, Parts A and B of Directive 1999/45/EC. In this case, the classification is based on the individual concentration limits

• in Annex I to Directive 67/548/EEC

• or in Annex III Part B to Directive 1999/45/EC where the substance or substances do not appear in Annex I to Directive 67/548/EEC or appear in it without concentration limits.

5.1.3. Normally, the classification of a preparation is made on the basis of a conventional method. However, for the determination of the acute aquatic toxicity, there may be cases for which it is appropriate to carry out tests on the preparation. The result of these tests on the preparation may only modify the classification concerning acute aquatic toxicity which would have been obtained by the application of a conventional method. If such tests are chosen by the person responsible for the placing on the market, it must be ensured that the quality criteria of the test methods in Part C of Annex V to Directive 67/548/EEC have been complied with. Furthermore, the tests are to be carried out on all three groups of species in conformity with the criteria in this Annex (algae, daphnia and fish), unless the highest hazard classification relating to acute aquatic toxicity has been assigned to the preparation after testing on one of the species or a test result was already available before Directive 1999/45/EC entered into force.

5.2. Criteria for classification, indication of danger, choice of risk phrases

The classification criteria for substances in Section 5.2.1. only apply to preparations where they have been tested in accordance with 5.1.3.

5.2.1. Aquatic environment

5.2.1.1. Substances shall be classified as dangerous for the environment and assigned the symbol ‘N’ and the appropriate indication of danger, and assigned risk phrases in accordance with the following criteria:

R50 Very toxic to aquatic organisms

and

R53 May cause long-term adverse effects in the aquatic environment

Acute toxicity: 96 hr LC50 (for fish)	≤ 1 mg/l
or 48 hr EC50 (for Daphnia)	≤ 1 mg/l
or 72 hr IC50 (for algae)	≤ 1 mg/l

and

• the substance is not readily degradable

or

• the log Pow (log octanol/water partition coefficient) ≥ 3.0 (unless the experimentally determined BCF ≤ 100).

R50 Very toxic to aquatic organisms

Acute toxicity: 96 hr LC50 (for fish)	≤ 1 mg/l
or 48 hr EC50 (for Daphnia)	≤ 1 mg/l
or 72 hr IC50 (for algae)	≤ 1 mg/l

R51 Toxic to aquatic organisms

and

R53 May cause long-term adverse effects in the aquatic environment

Acute toxicity: 96 hr LC50 (for fish)	1 mg/l < LC50 ≤ 10 mg/l
or 48 hr EC50 (for Daphnia)	1 mg/l < EC50 ≤ 10 mg/l
or 72 hr IC50 (for algae)	1 mg/l < IC50 ≤ 10 mg/l

and

• the substance is not readily degradable

or

• the log Pow ≥ 3.0 (unless the experimentally determined BCF ≤ 100).

5.2.1.2. Substances shall be classified as dangerous for the environment in accordance with the criteria set out below. Risk phrases shall also be assigned in accordance with the following criteria

R52 Harmful to aquatic organisms

and

R53 May cause long-term adverse effects in the aquatic environment

Acute toxicity: 96 hr LC50 (for fish)	1 mg/l < LC50 ≤ 100 mg/l
or 48 hr EC50 (for Daphnia)	1 mg/l < EC50 ≤ 100 mg/l
or 72 hr IC50 (for algae)	1 mg/l < IC50 ≤ 100 mg/l

and

the substance is not readily degradable.

This criterion applies unless there exists additional scientific evidence concerning degradation and/or toxicity sufficient to provide an adequate assurance that neither the substance nor its degradation products will constitute a potential long-term and/or delayed danger to the aquatic environment. Such additional scientific evidence should normally be based on the studies required at Level 1 (Annex VIII), or studies of equivalent value, and could include:

• (i) a proven potential to degrade rapidly in the aquatic environment,

• (ii) an absence of chronic toxicity effects at a concentration of 1.0 mg/litre, e.g. a no-observed effect concentration of greater than 1.0 mg/litre determined in a prolonged toxicity study with fish or Daphnia.

R52 Harmful to aquatic organisms

Substances not falling under the criteria listed above in this chapter, but which on the basis of the available evidence concerning their toxicity may nevertheless present a danger to the structure and/or functioning of aquatic ecosystems.

R53 May cause long-term adverse effects in the aquatic environment

Substances not falling under the criteria listed above in this chapter, but which, on the basis of the available evidence concerning their persistence, potential to accumulate, and predicted or observed environmental fate and behaviour may nevertheless present a long-term and/or delayed danger to the structure and/or functioning of aquatic ecosystems.

For example, poorly water-soluble substances, i.e. substances with a solubility of less than 1 mg/l will be covered by this criterion if:

• (a) they are not readily degradable; and

• (b) the log Pow ≥ 3.0 (unless the experimentally determined BCF ≤ 100).

This criterion applies to substances unless there exists additional scientific evidence concerning degradation and/or toxicity sufficient to provide an adequate assurance that neither the substance nor its degradation products will constitute a potential long-term and/or delayed danger to the aquatic environment.

Such additional scientific evidence should normally be based on the studies required at Level 1 (Annex VIII), or studies of equivalent value, and could include

• (i) a proven potential to degrade rapidly in the aquatic environment;

• (ii) an absence of chronic toxicity effects at the solubility limit e.g. a no-observed effect concentration of greater than the solubility limit determined in a prolonged toxicity study with fish or Daphnia.

5.2.1.3. Comments on the determination of IC50 for algae and of degradability

• where it can be demonstrated in the case of highly coloured substances that algal growth is inhibited solely as a result of a reduction in light intensity, then the 72h IC50 for algae should not be used as a basis for classification.

• Substances are considered readily degradable if the following criteria hold true.

• (a) If in 28-day biodegradation studies the following levels of degradation are achieved

• in tests based upon dissolved organic carbon: 70%,

• in tests based upon oxygen depletion or carbon dioxide generation: 60 % of the theoretical maxima.

These levels of biodegradation must be achieved within 10 days of the start of degradation, which point is taken as the time when 10% of the substance has been degraded.

or

• (b) if in those cases where only COD and BOD5 data are available when the ratio of BOD5/COD is greater than or equal to 0.5;

or

• (c) if other convincing scientific evidence is available to demonstrate that the substance can be degraded (biotically and/or abiotically) in the aquatic environment to a level of > 70% within a 28-day period.

5.2.2. Non - aquatic environment

5.2.2.1. Substances and preparations shall be classified as dangerous for the environment and assigned the symbol ‘N’ and the appropriate indication of danger, and assigned risk phrases in accordance with the following criteria:

R54 Toxic to flora

R55 Toxic to fauna

R56 Toxic to soil organisms

R57 Toxic to bees

R58 May cause long-term adverse effects in the environment

Substances and preparations which on the basis of the available evidence concerning their toxicity, persistence, potential to accumulate and predicted or observed environmental fate and behaviour may present a danger, immediate or long-term and/or delayed, to the structure and/or functioning of natural ecosystems other than those covered under 5.2.1 above. Detailed criteria will be elaborated later.

5.2.2.2. Substances and preparations shall be classified as dangerous for the environment, and assigned the symbol ‘N’ and the appropriate indication of danger, where applicable, and assigned risk phrases in accordance with the following criteria:

R59 Dangerous for the ozone layer

Substances which on the basis of the available evidence concerning their properties and their predicted or observed environmental fate and behaviour may present a danger to the structure and/or the functioning of the stratospheric ozone layer. This includes the substances which are listed in Annex I to Council Regulation (EC) No 2037/2000 on substances that deplete the ozone layer (OJ No L 244, 29.9.2000, p.1) and its subsequent amendments.

Preparations shall be classified on the basis of a conventional method referred to in Article 7 and Annex III, Parts A and B of Directive 1999/45/EC.

6.1. Introduction

Safety advice phrases (S-phrases) shall be assigned to dangerous substances and preparations in accordance with the following general criteria. In addition, for certain preparations, the safety advice listed in Annex V of Directive 1999/45/EC is mandatory.

Whenever the manufacturer is mentioned in Chapter 6 it refers to the person responsible for placing the substance or preparation on the market.

6.2. Safety phrases for substances and preparations

S1 Keep locked up

• Applicability:

• very toxic, toxic and corrosive substances and preparations.

• Criteria for use:

• obligatory for those substances and preparations mentioned above if sold to the general public.

S2 Keep out of the reach of children

• Applicability:

• all dangerous substances and preparations.

• Criteria for use:

• obligatory for all dangerous substances and preparations sold to the general public except for those only classified as dangerous for the environment.

S3 Keep in a cool place

• Applicability:

• organic peroxides,

• other dangerous substances and preparations having a boiling point ≤ 40° C.

• Criteria for use:

• obligatory for organic peroxides unless S47 is used,

• recommended for other dangerous substances and preparations having a boiling point ≤ 40°C.

S4 Keep away from living quarters

• Applicability:

• very toxic and toxic substances and preparations.

• Criteria for use: