Annex B 

Spirotetramat 

B.4 Proposal for classification and labelling 

Reviewed by: 

AGES, Austria 
– 2 – 

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

B.4.1 Proposals for the classification and labelling 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-term 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 stuffs 

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/face protection 

S56 Dispose of this material and its container to hazardous 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 special instructions/safety 

data sheets. 

Justification for the proposal 

Xn Applicable as R36, R43, R62 and R63 has been assigned. 

N Applicable as R51/53 has been assigned. 

R36 Corneal, iridal and conjunctival effects which persisted partly through day 7 of the study 

(Leuschner J., 2002) 

R43 Sensitization in the Magnusson and Kligman maximization 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 and skeletal deviations at maternal toxic 

doses (Klaus A. M., 2004) 

R51/53 In ecotoxicity tests with standard species (fish, daphnia 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 sensitization 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 sensitization by skin contact. 

S27/28 Spirotetramat may cause sensitization by skin contact. 

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 classification and labelling April 2008

Further explanation for classification and labelling concerning toxicology: 

R 62: 

In addition to testicular histopathology observed following subchronic and chronic exposure of male rats to 

spirotetramat, evidence of male reproductive toxicity was provided in the 2-generation reproductive toxicity 

study. Abnormal sperm cells were reported in F1-generation male rats treated with 6000 ppm (419 mg/kg 

bw/day) spirotetramat in the diet, and decreased reproductive performance was also observed in one of 

these males. Similar results were obtained in the 1-generation reproductive toxicity range-finding study, 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 no fertility in parental generation animals. There were no 

implantation sites noted in the females due to treatment-related effects on sperm cells of males at this dose 

level (increased numbers of abnormal sperms, reduced epididymal sperm counts, decline in both motility and 

progression of epididymal sperm cells). Absolute and relative weight of the cauda epididymis was decreased 

in parental males. Histopathology showed abnormal sperm 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 paper are: 

• Only in rats and only at very high doses the male reproductive system was affected (reproductive 

performance, sperm count, morphology and motility, weight of cauda epididymis, testicular and/or 

epididymal morphology). This is a high-dose phenomenon with a steep dose-response curve and 

reversibility after cessation of exposure. The overall 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 reproductive system in the subchronic and chronic studies 

even at doses exceeding the limit dose of 1000 mg/kg bw/day. 

• From multiple special mechanistic studies, the pathomechanism and toxicokinetics in rats and the 

relevance for humans are well understood: 

1. More than 10 days of treatment with repeated daily 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 dysregulation. 

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 special 

toxicokinetic features: Due to the anionic state of the enol, hepatic and renal elimination of BYI 08330-

enol occur via very efficient active transport processes (against a concentration gradient) which can 

only be overwhelmed (saturated) by very high (approx. 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 study using hepatocytes from male rats, mice and 

humans revealed species differences in the proportion of spirotetramat metabolites formed. 

Specifically, mouse hepatocytes were better able than rat liver cells to metabolize BYI 08330-enol via

glucuronidation. The metabolic profile generated from the human hepatocytes was more comparable 
– 4 – 

Spirotetramat – Volume 3, Annex B.4, Proposal for classification 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 transformation noted in rats. Results obtained from rat 

hepatocytes corresponded to in vivo metabolism findings in the rat. 

8. This efficient metabolic elimination in mice is the obvious reason for the absence of testicular toxicity 

even at doses >1000 mg/kg bw. Based on these metabolic features, it can be expected that humans 

are, similar to mice, less sensitive than rats to effects of BYI 08330 on the male reproductive system. 

9. The rat is therefore not the relevant species for human risk assessment. But even if humans would be 

as sensitive as rats, humans will not develop testicular toxicity under practical conditions (>6000-fold 

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 repeated-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-finding study, decreased sperm motility and progression 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 generation 

animals. 

The NOAEL for reproductive toxicity was 70 mg/kg bw/day. So the effects on the male reproductive system 

are no “high dose phenomenon”.

The effects on testicular spermatogenesis were attributed to the BYI 08330-enol, because it is the main 

metabolite in the rat. But it can not be excluded that other metabolites can cause the same effects. 

BYI 08330-enol is further metabolised by oxidation reactions to BYI 08330-desmethylenol, BYI 08330-enolalcohol and BYI 08330-ketohydroxy. Oxidation products accounted for approximately 14%. Conjugation was

not detected. In the mouse, conjugation of BYI 08330-enol with glucuronic acid accounted for approximately 

30%. In human hepatozytes, conjugation to BYI 08330-enol-glucuronic acid was 6%. The in vitro conjugation 

rate is dependent on the concentration used and declined in mice from 30% to 9% and in humans from 6% 

to 2% at liver concentrations of 19 μg/g and 190 μg/g BYI 08330, respectively. Glucuronidation of the BYI 

08330-enol in mice leads to much lower systemic levels of free BYI 08330-enol when compared to the rat. 

The conjugation enables the mouse to utilize separate active transport systems in the kidneys, thus avoiding 

a saturation of the elimination process. Thus, the utilization of different transport systems renders the mouse 

less sensitive to BYI 08330-mediated testicular toxicity 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 mouse 

(dependent on the concentration, in humans 6% respectively 2%, in mice 30% respectively 9%). Therefore a 

similarity in the metabolic pathway can not be followed. As for humans conjugation is only 2% at high doses, 

it can not be assumed that humans are less sensitive to Spirotetramat than rats. 

Due to abnormal sperm cells, decreased sperm motility 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 fertility“ according to Annex VI of the EC Council Directive 

67/548/EEC. 

R 63: 

In the developmental toxicity study in rats, toxicity 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 fetal 

weight and increased incidences of skeletal malformations and skeletal deviations were observed at 1000 
– 5 – 

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

mg/kg bw/day. Malformations at the high dose included one case of supernumerary lumbar vertebra, one 

case of cleft palate and one case of co-arctation of aortic arch. One case of atrial septal defect of the heart 

and microphthalmia were observed in the control, low 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 with 

pelvic shift (1.1 %) were observed in the high dose. Historical control data of the performing laboratory 

(Bayer HealthCare AG) for dysplastic forelimb bones in studies conducted in the years 1999 – 2004 showed 

26 affected animals out of 1975 animals, a percentage 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 concurrent 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 in 

historical controls in studies conducted 1999 – 2006 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 compared to 

concurrent and historical control values . Statistically significantly increased numbers of fetuses with 14th ribs 

were observed at a dose level of 1000 mg/kg/d spirotetramat. 

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] 

14th rib 15 10.7 12.6 45.9** (0.0 – 13) 

[1996 – 2000] 

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

Due to increased incidences of skeletal malformations and skeletal deviations and according to Annex VI 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 – Possible risk of harm to the unborn child“. 
– 6 – 

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

B.4.2 Proposals for the classification and labelling of the preparation (Annex IIIA 12.3 and 12.4) 

On the basis of the available data the following classification 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-term 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 stuffs 

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/face protection 

S56 Dispose of this material and its container to hazardous 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 special instructions/safety 

data sheets. 

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

clean application equipments near surface water/Avoid contamination 

via drains from farmyards and roads 

SPe3 To protect aquatic organisms respect an unsprayed buffer zone of 5 m 

(citrus) to surface water bodies 

SPe8 Dangerous to bees. To protect bees and other pollinating insects do not 

apply to crop plants when in flower or when flowering 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 and skeletal deviations at maternal toxic 

doses (Klaus A. M., 2004) 

R51/53 In ecotoxicity tests with standard species (fish and aquatic invertebrates), Onchorynchus 

mykiss and Chironomus riparius are the most sensitive species with a EC50 value of 

9.48 mg/l (Nieden 2005b) and 5.44 mg/L (Dorgerloh 2006d), respectively. The active 

substance spirotetramat is not readily biodegradable (Weyers, 2005). 

S2, S13, S20/21, 

S36/37/39 Spirotetramat may cause sensitization by skin contact and has possible risks of impaired 

fertility and harm to the unborn child. 

S24, S27/28 Spirotetramat may cause sensitization by skin contact. 
– 7 – 

Spirotetramat – Volume 3, Annex B.4, Proposal for classification 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 required to achieve a safe use in citrus. 

Spe8 Transient effects on honey bee larvae have been observed in semi filed trials, therefore 

precautionary risk mitigation measures are recommended. 
– 8 – 

Spirotetramat – Volume 3, Annex B.4, Proposal for classification 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 study 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 potential 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 classification 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 08330 - 

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 developmental 

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 08330 - 

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 - Investigation 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 classification 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