# Annex B



# Spirotetramat

B.4 Proposal for classification and labelling

Reviewed by:

AGES, Austria– 2 –

Spirotetramat – Volume 3, Annex B.4, Proposal for c lassification and labelling April 2008

B.4.1 Proposals for the classification and labellin g of the active substance (Annex IIA 10)

Classification proposed is in accordance with Annex VI to Directive 67/548/EEC.

Hazard symbol(s)

Indications of danger Xn Harmful N Dangerous for the environment Risk phrases R36 Irritating to eyes R43 May cause sensitisation by skin contact R62 Possible risk of impaired fertility R63 Possible risk of harm to the unborn child R51/53 Toxic to aquatic organisms, may cause long-t erm adverse effect in the

aquatic environment. Safety phrases S2 Keep out of the reach of children S13 Keep away from food, drink and animal feeding stu ffs S20/21 When using do not eat, drink or smoke S24/25 Avoid contact with skin and eyes S27/28 After contact with skin, take off immediately all contaminated clothing

and wash immediately with plenty of water. S36/37/39 Wear suitable protective clothing, gloves and eye/f ace protection S56 Dispose of this material and its container to ha zardous or special waste

collection point. S57 Use appropriate container to avoid environmental contamination. S60 This material and its container must be disposed of as hazardous

waste. S61 Avoid release to the environment. Refer to specia l instructions/safety

data sheets. Justification for the proposal

Xn Applicable as R36, R43, R62 and R63 has been assign ed. N Applicable as R51/53 has been assigned. R36 Corneal, iridal and conjunctival effects which pe rsisted partly through day 7 of the study

(Leuschner J., 2002) R43 Sensitization in the Magnusson and Kligman maximi zation study and the Local lymph node

assay (Vohr H. W., 2002; Esdaile D., 2004) R62 Evidence of male reproductive toxicity (Young A., 2006) R63 Increased incidences of skeletal malformations an d skeletal deviations at maternal toxic

doses (Klaus A. M., 2004) R51/53 In ecotoxicity tests with standard species (fish, d aphnia and algae), Lepomis macrochirus is the most sensitive species with a EC50 value of 2.2 mg/l (Dorgerloh 2005a). Spirotetramat is not readily biodegradable (Weyers, 2005). S2, S13, S20/21, S36/37/39 Spirotetramat is irritating to eyes, may cause sens itization by skin contact and has possible

risks of impaired fertility and harm to the unborn child. S24/25 Spirotetramat is irritating to eyes and may cause s ensitization by skin contact. S27/28 Spirotetramat may cause sensitization by skin conta ct. S56 and S57 Because of the toxicity of spirotetramat to aquatic organisms and potential long term

adverse effects to the aquatic environment S60 Applicable to all dangerous substances. S61 Applicable as R51/53 has been assigned.– 3 –

Spirotetramat – Volume 3, Annex B.4, Proposal for c lassification and labelling April 2008

Further explanation for classification and labellin g concerning toxicology:

R 62: In addition to testicular histopathology observed f ollowing subchronic and chronic exposure of male ra ts to spirotetramat, evidence of male reproductive toxici ty was provided in the 2-generation reproductive to xicity study. Abnormal sperm cells were reported in F1-gen eration male rats treated with 6000 ppm (419 mg/kg bw/day) spirotetramat in the diet, and decreased re productive performance was also observed in one of these males. Similar results were obtained in the 1 -generation reproductive toxicity range-finding stu dy, in which decreased sperm motility and progression and increased abnormal sperm cells in the epididymides were observed in F1 males at ≥ 6000 ppm (320 mg/kg bw/day. The highest dose level of 10000 ppm, equivalent to 538 mg/kg bw/d, was associated with n o fertility in parental generation animals. There w ere no implantation sites noted in the females due to trea tment-related effects on sperm cells of males at th is dose level (increased numbers of abnormal sperms, reduce d epididymal sperm counts, decline in both motility and progression of epididymal sperm cells). Absolute an d relative weight of the cauda epididymis was decre ased in parental males. Histopathology showed abnormal s perm cells of minimal to moderate severity in the epididymis and the cauda epididymis.

In February 2008 the notifier submitted a position paper ( High dose reproductive effects in male rats and their relevance to humans; Temerowski M., 2008 ).The main statements and conclusions from this pap er are:

• Only in rats and only at very high doses the male r eproductive system was affected (reproductive performance, sperm count, morphology and motility, weight of cauda epididymis, testicular and/or epididymal morphology). This is a high-dose phenome non with a steep dose-response curve and reversibility after cessation of exposure. The over all NOAEL for these effects in the reproductive, subchronic and chronic studies is 320 mg/kg bw/day.

• Mice are insensitive to effects on the male reprodu ctive system in the subchronic and chronic studies even at doses exceeding the limit dose of 1000 mg/k g bw/day.

• From multiple special mechanistic studies, the path omechanism and toxicokinetics in rats and the relevance for humans are well understood:
1. More than 10 days of treatment with repeated dai ly doses of 1000 mg BYI 08330 per kg bw were needed to cause spermatid degeneration in rats.
2. There was no evidence of a hormonal dysregulatio n. 3. These effects are obviously caused by disturbed metabolic support of germ cells by Sertoli cells.
4. As mentioned above, these effects are reversible after cessation of exposure.
5. These effects were caused by BYI 08330-enol, the main rat metabolite which predominates in blood ( ~ 90%;) at physiological pH in the anionic form (pKa of enol = 5.2).
6. The low toxic potency (high-dose phenomenon) and the delayed onset of effects are related to specia l toxicokinetic features: Due to the anionic state of the enol, hepatic and renal elimination of BYI 083 30- enol occur via very efficient active transport proc esses (against a concentration gradient) which can only be overwhelmed (saturated) by very high (appro x. 500 mg/kg bw/day) and repeated doses. Lower-dose (<320 mg/kg bw day) and short-term (<10 days) exposure can be efficiently handled by the active elimination.
7. Results from a comparative in vitro metabolism s tudy using hepatocytes from male rats, mice and humans revealed species differences in the proporti on of spirotetramat metabolites formed. Specifically, mouse hepatocytes were better able th an rat liver cells to metabolize BYI 08330-enol via glucuronidation. The metabolic profile generated fr om the human hepatocytes was more comparable– 4 –

Spirotetramat – Volume 3, Annex B.4, Proposal for c lassification and labelling April 2008

to that of the mouse than of the rat. In both mouse and human tissue, conjugation was prevalent compared to the unexceptional oxidative transformat ion noted in rats. Results obtained from rat hepatocytes corresponded to in vivo metabolism find ings in the rat.

8. This efficient metabolic elimination in mice is the obvious reason for the absence of testicular to xicity even at doses >1000 mg/kg bw. Based on these metabo lic features, it can be expected that humans are, similar to mice, less sensitive than rats to e ffects of BYI 08330 on the male reproductive system

. 9. The rat is therefore not the relevant species fo r human risk assessment. But even if humans would b e as sensitive as rats, humans will not develop testi cular toxicity under practical conditions (>6000-fo ld margin of safety between rat testicular NOAEL (320 mg/kg bw/day) and AOEL / ADI (0.05 mg/kg bw/day)). And even high-dose accidental exposure or self-poisoning will not cause testicular toxicity in humans due to the high-dose phenomenon, the repeate d-dose prerequisite, and the recovery of testicular toxicity: Additional Note: To reach the experimental NOAEL for testicular toxicity in rats, a 70 kg person would have to ingest 22,000 mg BYI 08330 for more than 10 consecutive days.

10. It is therefore not justified to classify BYI 08330 to any category of reproductive effects.

Comment (RMS): The statement of the notifier can not be agreed to. In the 1-generation reproductive toxicity range-fin ding study, decreased sperm motility and progressio n and increased abnormal sperm cells in the epididymides were observed in F1 males at ≥ 320 – 400 mg/kg bw/day and the highest dose level of 538 mg/kg bw/d was associated with no fertility in parental gener ation animals. The NOAEL for reproductive toxicity was 70 mg/kg bw /day. So the effects on the male reproductive syste m

are no “high dose phenomenon”. The effects on testicular spermatogenesis were attr ibuted to the BYI 08330-enol, because it is the mai n metabolite in the rat. But it can not be excluded t hat other metabolites can cause the same effects. BYI 08330-enol is further metabolised by oxidation reactions to BYI 08330-desmethylenol, BYI 08330-eno lalcohol and BYI 08330-ketohydroxy. Oxidation produc ts accounted for approximately 14%. Conjugation was not detected. In the mouse, conjugation of BYI 0833 0-enol with glucuronic acid accounted for approxima tely 30%. In human hepatozytes, conjugation to BYI 08330 -enol-glucuronic acid was 6%. The in vitro conjugat ion rate is dependent on the concentration used and dec lined in mice from 30% to 9% and in humans from 6% to 2% at liver concentrations of 19 μ g/g and 190 μ g/g BYI 08330, respectively. Glucuronidation of the BYI 08330-enol in mice leads to much lower systemic lev els of free BYI 08330-enol when compared to the rat

. The conjugation enables the mouse to utilize separa te active transport systems in the kidneys, thus av oiding a saturation of the elimination process. Thus, the utilization of different transport systems renders the mouse less sensitive to BYI 08330-mediated testicular tox icity when compared to the rat. In contrast to mice , for humans the ability to conjugate BYI 08330-enol with glucuronic acid is fivefold lower than for the mou se (dependent on the concentration, in humans 6% respe ctively 2%, in mice 30% respectively 9%). Therefore a similarity in the metabolic pathway can not be foll owed. As for humans conjugation is only 2% at high doses, it can not be assumed that humans are less sensitiv e to Spirotetramat than rats.

Due to abnormal sperm cells, decreased sperm motili ty and progression and decreased reproductive performance, spirotetramat should be classified to “category 3 of reproductive substances “ and labelled with the risk phrase “R 62 – Possible risk of impaired f ertility“ according to Annex VI of the EC Council D irective 67/548/EEC.

R 63: In the developmental toxicity study in rats, toxici ty to the offspring was observed in the presence of maternal toxicity, including decreased food consumption and body weight/gain, at 1000 mg/kg bw/day. Reduced fet al weight and increased incidences of skeletal malform ations and skeletal deviations were observed at 100 0– 5 –

Spirotetramat – Volume 3, Annex B.4, Proposal for c lassification and labelling April 2008

mg/kg bw/day. Malformations at the high dose includ ed one case of supernumerary lumbar vertebra, one case of cleft palate and one case of co-arctation o f aortic arch. One case of atrial septal defect of the heart and microphthalmia were observed in the control, lo w and high dose each, but not at the mid dose. Four cases of dysplastic forelimb bones (1.5 %) and three cases of malformed sacral vertebral arches w ith pelvic shift (1.1 %) were observed in the high dose . Historical control data of the performing laborat ory (Bayer HealthCare AG) for dysplastic forelimb bones in studies conducted in the years 1999 – 2004 show ed 26 affected animals out of 1975 animals, a percenta ge of 1.3 % [range 0.4 – 4.3% due to one study conducted in the year 2000, were 10 animals out of 232 were affected (4.3%)]. An incidence of 1.5 % in the study with spirotetramat is therefore outside the c oncurrent control (0.4%) and the historical control data (1.3%).

Statistically significantly increased incidences of sacral vertebral alterations (1.1 %) were observed at a dose level of 1000 mg/kg bw spirotetramat in comparison to the concurrent controls (0.0 %). The incidence i n historical controls in studies conducted 1999 – 200 6 showed 2 affected animals out of 6554 animals, a percentage of 0.03 % (range 0 – 0.4%).

Percentage of main skeletal observations in the rat developmental toxicity study with Spirotetramat (%)
Dose (mg/kg/d)

Observations 0 20 140 1000 % of Historical control data

Dysplasia of forelimb bones (%) 0.4 0.7 0.0 1.5** 1.3 (26 out of 1975 animals) [1999 – 2004] Altered appearance of sacral vertebral arch; pelvic shift (%) 0.0 0.0 0.0 1.1** 0.03 (2 out of 6554 animals) [1999 – 2006]
* p<0.05; **P<0.01 (Fisher’s exact test)

Statistically significantly increased incidences of wavy ribs were observed at all dose levels compare d to concurrent and historical control values . Statisti cally significantly increased numbers of fetuses wi th 14 th ribs were observed at a dose level of 1000 mg/kg/d spiro tetramat.

Percentage of observations at the ribs
Dose (mg/kg/d)

Observations 0 20 140 1000 Historical control data (range)

Wavy ribs 5.5 17** 17.8** 57.5** (2.7 – 15.1) [1996 – 2001]

14 th rib 15 10.7 12.6 45.9** (0.0 – 13) [1996 – 2000]

The maternal and the developmental NOAEL for this s tudy was set at 140 mg/kg bw/d.

Due to increased incidences of skeletal malformatio ns and skeletal deviations and according to Annex V I of the EC Council Directive 67/548/EEC, spirotetramat should be classified to “category 3 of reproductive substances “ and labelled with the risk phrase “R 63 – Possibl e risk of harm to the unborn child“.– 6 –

Spirotetramat – Volume 3, Annex B.4, Proposal for c lassification and labelling April 2008

B.4.2 Proposals for the classification and labellin g of the preparation (Annex IIIA 12.3 and 12.4) On the basis of the available data the following cl assification and labelling is proposed according to Directive 1999/45/EC:

Hazard symbol(s)

Indications of danger Xn Harmful N Dangerous for the environment Risk phrases R43 May cause sensitisation by skin contact R62 Possible risk of impaired fertility R63 Possible risk of harm to the unborn child R51/53 Toxic to aquatic organisms, may cause long-t erm adverse effect in the

aquatic environment. Safety phrases S2 Keep out of the reach of children S13 Keep away from food, drink and animal feeding stu ffs S20/21 When using do not eat, drink or smoke S24 Avoid contact with skin S27/28 After contact with skin, take off immediately all contaminated clothing

and wash immediately with plenty of water. S36/37/39 Wear suitable protective clothing, gloves and eye/f ace protection S56 Dispose of this material and its container to ha zardous or special waste

collection point. S57 Use appropriate container to avoid environmental contamination. S60 This material and its container must be disposed of as hazardous

waste. S61 Avoid release to the environment. Refer to speci al instructions/safety

data sheets. SP1 Do not contaminate water with the product or its containers (Do not

clean application equipments near surface water/Avoi d contamination via drains from farmyards and roads SPe3 To protect aquatic organisms respect an unspraye d buffer zone of 5 m

(citrus) to surface water bodies SPe8 Dangerous to bees. To protect bees and other p ollinating insects do not

apply to crop plants when in flower or when floweri ng weeds are present. Do not use when bees are actively foraging

Justification for the proposal:

Xn Applicable as R43, R62 and R63 has been assigned. N Applicable as R51/53 has been assigned. R43 Sensitization in the Buehler Patch Test (Vohr H. W., 2005) R62 Evidence of male reproductive toxicity (Young A., 2006) R63 Increased incidences of skeletal malformations an d skeletal deviations at maternal toxic

doses (Klaus A. M., 2004) R51/53 In ecotoxicity tests with standard species (fish an d aquatic invertebrates), Onchorynchus

mykiss and Chironomus riparius are the most sensitive species with a EC50 value o f 9.48 mg/l (Nieden 2005b) and 5.44 mg/L (Dorgerloh 2 006d), respectively. The active substance spirotetramat is not readily biodegradabl e (Weyers, 2005). S2, S13, S20/21, S36/37/39 Spirotetramat may cause sensitization by skin conta ct and has possible risks of impaired

fertility and harm to the unborn child. S24, S27/28 Spirotetramat may cause sensitization by skin conta ct.– 7 –

Spirotetramat – Volume 3, Annex B.4, Proposal for c lassification and labelling April 2008

S56 and S57 Because of the toxicity of spirotetramat to aquatic organisms and potential long term

adverse effects to the aquatic environment S60 Applicable to all dangerous substances S61 Applicable as R51/53 has been assigned SP1 Mandatory for all plant-protection products SPe3 Risk mitigation measures (buffer zone) are requir ed to achieve a safe use in citrus. Spe8 Transient effects on honey bee larvae have been o bserved in semi filed trials, therefore

precautionary risk mitigation measures are recommen ded.– 8 –

Spirotetramat – Volume 3, Annex B.4, Proposal for c lassification and labelling April 2008

References relied on

Annex point/
reference
number Author(s) Year Title Source (where different from company) Company, Report No GLP or GEP status (where relevant)
Published or not Data
Protection Claimed

Y/N-R/NR Owner

Annex II Data and Information IIA, 5.2.5 Leuschner, P 2002 Acute eye irritation stu dy of BYI 08330 by instillation into the conjunctival sac of rabbits LPT Laboratory of Pharmacology and Toxicology KG, Hamburg, Germany Bayer CropScience AG, Report No.: R8146, Edition Number: M-062864-01-3 Date: 25.04.2002 GLP, unpublished Y BCS

IIA, 5.2.6 Vohr, H 2002 BYI 08330 - Study for the skin sensitization effect in guinea pigs (guinea pig maximization test according to Magnusson and Kligman) Bayer AG, Wuppertal, Germany Bayer CropScience AG, Report No.: 32273, Edition Number: M-076253-01-2 Date: 29.07.2002 GLP, unpublished Y BCS

IIA, 5.2.6 Esdaile, D 2004 BYI 08330 - Evaluation of po tential dermal sensitization in the local lymph node assay Bayer CropScience SA, Sophia Antipolis, France Bayer CropScience AG, Report No.: SA 04120, Edition Number: M-090707-01-2 Date: 09.09.2004 GLP, unpublished Y BCS

IIA, 5.6.1 Young, A.D. 2006 Technical grade BY1 08330 - A dose range-finding reproductive toxicity study in the Wistar rat (revised report) Bayer CropScience, Stilwell, KS, USA Bayer CropScience AG, Report No.: 201300-1, Edition Number: M-273578-02-1 Date: 30.05.2006, Amended: 12.07.2006 GLP, unpublished Y BCS

IIA, 5.6.1 Young, A.D. 2006 Technical grade BYI 08330 (common name Spirotetramat): A two generation reproductive toxicity study in the Wistar rat Bayer CropScience, Stilwell, KS, USA Bayer CropScience AG, Report No.: 201426-1, Edition Number: M-274619-02-1 Date: 30.05.2006, Amended: 13.07.2006 GLP, unpublished Y BCS– 9 –

Spirotetramat – Volume 3, Annex B.4, Proposal for c lassification and labelling April 2008

Annex point/
reference
number Author(s) Year Title Source (where different from company) Company, Report No GLP or GEP status (where relevant)
Published or not Data
Protection Claimed

Y/N-R/NR Owner

IIA, 5.6.2 Klaus, A.M. 2004 BYI 08330, Synonym: FHN 0833 0 - Developmental toxicity study in rats after oral administration Bayer HealthCare AG, Wuppertal, Germany Bayer CropScience AG, Report No.: AT01413, Edition Number: M-086404-01-2 Date: 23.08.2004 GLP, unpublished Y BCS

IIA, 5.6.2 Klaus, A.M. 2001 BYI 08330 - Pilot study on d evelopmental toxicity in rats after oral administration Bayer AG, Wuppertal, Germany Bayer CropScience AG, Report No.: T3068559, Edition Number: M-021476-01-2 Date: 08.05.2001 Non GLP, unpublished Y BCS

IIA, 5.6.2 Klaus, A.M. 2004 BYI 08330 - Synonym: FHN 083 30 - Supplementary developmental toxicity study in rats after oral administration Bayer HealthCare AG, Wuppertal, Germany Bayer CropScience AG, Report No.: AT01512, Edition Number: M-091750-01-2 Date: 07.10.2004 GLP, unpublished Y BCS

IIA, 5.6.2 Temerowski, M. 2008 Spirotetramat (syn. BYI 08330) - Request for historical control data Bayer CropScience AG, Report No.: M-296860-01-1, Edition Number: M-296860-01-1 Date: 30.01.2008 Non GLP, unpublished Y BCS

IIA, 5.8.1 Kennel, P 2005 BYI 08330 - Evaluation of the potential reproductive toxicity in the male rat following daily oral administration by gavage Bayer CropScience SA, Sophia Antipolis, France Bayer CropScience AG, Report No.: SA 04181, Edition Number: M-252001-01-2 Date: 23.05.2005 GLP, unpublished Y BCS

IIA, 5.8.1 Tinwell, H 2006 BYI 08330-Enol - Investigat ion of the testicular/sperm toxicity in the rat following 21 days of exposure by gavage Bayer CropScience SA, Sophia Antipolis, France Bayer CropScience AG, Report No.: SA06011, Edition Number: M-273959-01-1 Date: 30.06.2006 Non GLP, unpublished Y BCS– 10 –

Spirotetramat – Volume 3, Annex B.4, Proposal for c lassification and labelling April 2008

Annex point/
reference
number Author(s) Year Title Source (where different from company) Company, Report No GLP or GEP status (where relevant)
Published or not Data
Protection Claimed

Y/N-R/NR Owner

IIA, 5.8.1 Temerowski, M. 2008 Spirotetramat (syn. BYI 08330) - High dose reproductive effects in male rats and their relevance to humans Bayer CropScience AG, Report No.: M-297775-01-1, Edition Number: M-297775-01-1 Date: 20.02.2008 Non GLP, unpublished Y BCS

IIA, 7.7 Weyers, A. 2005 BYI 08330 - Biodegradation Bayer Industry Services, Leverkusen, Germany Bayer CropScience AG, Report No.: 2005/0077/01, Edition Number: M-263287-01-1 Date: 16.11.2005 GLP, unpublished Yes BCS

IIA 8.2.1.2 Dorgerloh, M. 2005

a Acute toxicity of BYI 08330 (tech.) to fish (Lepomis macrochirus) Bayer CropScience AG, Report No.: DOM 24056, Edition Number: M-242689-01-2 Date: 12.01.2005 GLP unpublished Yes BCS

Annex III Data and Information IIIA, 7.1.6 Vohr, H 2005 BYI 08330 150 OD (Project: BYI 08330) - Study for the skin sensitization effect in guinea pigs (Buehler patch test) Bayer HealthCare AG, Wuppertal, Germany Bayer CropScience AG, Report No.: AT01873, Edition Number: M-246231-02-2 Date: 18.02.2005 GLP, unpublished also filed: KIIIA2 7.1.6 /01 Y BCS

IIIA, 10.2.2.1 Nieden, D. 2005 d Acute toxicity of BYI 08330 OD 150 to fish ( Oncorhynchus mykiss ) under static conditions Bayer CropScience GmbH, Monheim, Germany Bayer CropScience AG, Report No.: EBFNM008, Edition Number: M-253916-01-2 Date: 29.06.2005 GLP, unpublished Yes BCS

IIIA, 10.2.2.2 Dorgerloh, M. 2006 d Acute toxicity of BYI 08330 OD 150 to larvae of Chironomus riparius in a 48 h static laboratory test system Bayer CropScience AG, Report No.: EBFNM007, Edition Number: M-264018-01-2 Date: 19.01.2006 GLP, unpublished Yes BCS