margosa extract product-type 18 (insecticides, acaricides ... · margosa extract product-type 18...

Directive 98/8/EC concerning the placing biocidal

products on the market

Inclusion of active substances in Annex I or IA to Directive 98/8/EC

Assessment Report

Margosa Extract Product-type 18 (Insecticides, Acaricides and Products to

control other Arthropods)

09/12/2011

Annex I - Germany

Margosa Extract Product-type 18 09/12/2011

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Margosa Extract (PT18)

Assessment report

Finalised in the Standing Committee on Biocidal Products at its meeting on 09/12/2011 in

view of its inclusion in Annex I to Directive 98/8/EC

CONTENTS

1. STATEMENT OF SUBJECT MATTER AND PURPOSE .................................. 4

1.1. Procedure followed.......................................................................................... 4

1.2. Purpose of the assessment report ................................................................... 5

1.3. Overall conclusion in the context of Directive 98/8/EC ............................... 5

2. OVERALL SUMMARY AND CONCLUSIONS................................................... 7

2.1. Presentation of the Active Substance............................................................. 7

2.1.1. Identity, Physico-Chemical Properties & Methods of Analysis....... 7

2.1.2. Intended Uses and Efficacy................................................................ 8

2.1.3. Classification and Labelling .............................................................. 9

2.2. Summary of the Risk Assessment ................................................................ 10

2.2.1. Human Health Risk Assessment...................................................... 10

2.2.1.1. Hazard identification ......................................................................... 10

2.2.1.2. Effects assessment ............................................................................. 15

2.2.1.3. Exposure assessment ......................................................................... 15

2.2.1.4. Risk characterisation ......................................................................... 18

2.2.2. Environmental Risk Assessment...................................................... 24

2.2.2.1. Fate and distribution in the environment........................................... 24

2.2.2.2. Effects assessment ............................................................................. 26

2.2.2.3. PBT assessment ................................................................................. 28

2.2.2.4. Exposure assessment ......................................................................... 29

2.2.2.5. Risk characterisation ......................................................................... 31

2.2.3. List of endpoints ............................................................................... 33

3. DECISION ............................................................................................................... 34

3.1. Background to the Decision.......................................................................... 34

3.2. Decision regarding Inclusion in Annex I..................................................... 35


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3.3. Elements to be taken into account by Member States when authorising

products .......................................................................................................... 36

3.4. Requirement for further information.......................................................... 37

3.5. Updating this Assessment Report ................................................................ 37

APPENDIX I: LIST OF ENDPOINTS ......................................................................... 38

APPENDIX II: LIST OF INTENDED USES ............................................................... 63

APPENDIX III: LIST OF STUDIES............................................................................. 64


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1. STATEMENT OF SUBJECT MATTER AND PURPOSE

1.1. Procedure followed

This assessment report has been established as a result of the evaluation of margosa extract as product-type 18 (Insecticides, Acaricides and Products to control other Arthropods), carried out in the context of the work programme for the review of existing active substances provided for in Article 16(2) of Directive 98/8/EC concerning the placing of biocidal products on the market1, with a view to the possible inclusion of this substance into Annex I or IA to the Directive.

Margosa extract (CAS no. 84696-25-3) was notified as an existing active substance, by Trifolio-M GmbH, hereafter referred to as the applicant, in product-type 18.

Regulation (EC) No 1451/2007 of 4 December 2007,2 which has repealed and replaced Commission Regulation (EC) No 2032/2003 of 4 November 2003,3 lays down the detailed rules for the evaluation of dossiers and for the decision-making process in order to include or not an existing active substance into Annex I or IA to the Directive.

In accordance with the provisions of Article 5(2) of Regulation (EC) No 2032/2003, Germany was designated as Rapporteur Member State to carry out the assessment on the basis of the dossier submitted by the applicant. The deadline for submission of a complete dossier for margosa extract as an active substance in Product Type 18 was 30.04.2006, in accordance with Annex V of Regulation (EC) No 2032/2003.

On 02.05.2006, German competent authorities received a dossier from the applicant. The Rapporteur Member State accepted the dossier as complete for the purpose of the evaluation on 10.11.2006.

On 26.11.2009, the Rapporteur Member State submitted, in accordance with the provisions of Article 14(4) and (6) of Regulation (EC) No 1451/2007, to the Commission and the applicant a copy of the evaluation report, hereafter referred to as the competent authority report. The Commission made the report available to all Member States by electronic means on 08.01.2010. The competent authority report included a recommendation for the inclusion of margosa extract in Annex I to the Directive for PT18.

1 Directive 98/8/EC of the European Parliament and of the Council of 16 February 1998 concerning the placing biocidal products on the market. OJ L 123, 24.4.98, p.1

2 Commission Regulation (EC) No 1451/2007 of 4 December 2007 on the second phase of the 10-year work programme referred to in Article 16(2) of Directive 98/8/EC of the European Parliament and of the Council concerning the placing of biocidal products on the market. OJ L 325, 11.12.2007, p. 3

3 Commission Regulation (EC) No 2032/2003 of 4 November 2003 on the second phase of the 10-year work programme referred to in Article 16(2) of Directive 98/8/EC of the European Parliament and of the Council concerning the placing of biocidal products on the market and amending Regulation (EC) No 1896/2000. OJ L 307, 24.11.2003, p. 1


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In accordance with Article 16 of Regulation (EC) No 1451/2007, the Commission made the competent authority report publicly available by electronic means on 12.01.2010. This report did not include such information that was to be treated as confidential in accordance with Article 19 of Directive 98/8/EC.

In order to review the competent authority report and the comments received on it, consultations of technical experts from all Member States (peer review) were organised by the Commission. Revisions agreed upon were presented at technical and competent authority meetings and the competent authority report was amended accordingly.

On the basis of the final competent authority report, the Commission proposed the inclusion of margosa extract in Annex I to Directive 98/8/EC and consulted the Standing Committee on Biocidal Product on 09/12/2011.

In accordance with Article 15(4) of Regulation (EC) No 1451/2007, the present assessment report contains the conclusions of the Standing Committee on Biocidal Products, as finalised during its meeting held on 09/12/2011.

1.2. Purpose of the assessment report

This assessment report has been developed and finalised in support of the decision to include margosa extract in Annex I to Directive 98/8/EC for product-type 18. The aim of the assessment report is to facilitate the authorisation in Member States of individual biocidal products in product-type 18 that contain margosa extract. In their evaluation, Member States shall apply the provisions of Directive 98/8/EC, in particular the provisions of Article 5 as well as the common principles laid down in Annex VI.

For the implementation of the common principles of Annex VI, the content and conclusions of this assessment report, which is available at the Commission website4, shall be taken into account.

However, where conclusions of this assessment report are based on data protected under the provisions of Directive 98/8/EC, such conclusions may not be used to the benefit of another applicant, unless access to these data has been granted.

1.3. Overall conclusion in the context of Directive 98/8/EC

The overall conclusion from the evaluation is that it may be expected that there are products containing margosa extract for the product-type 18, which will fulfil the requirements laid down in Article 10(1) and (2) of Directive 98/8/EC. This conclusion is however subject to:

i. compliance with the particular requirements in the following sections of this assessment report,

4 http://ec.europa.eu/comm/environment/biocides/index.htm


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ii. the implementation of the provisions of Article 5(1) of Directive 98/8/EC, and

iii. the common principles laid down in Annex VI to Directive 98/8/EC.

Furthermore, these conclusions were reached within the framework of the uses that were proposed and supported by the applicant (see Appendix II). Extension of the use pattern beyond those described will require an evaluation at product authorisation level in order to establish whether the proposed extensions of use will satisfy the requirements of Article 5(1) and of the common principles laid down in Annex VI to Directive 98/8/EC.


2. OVERALL SUMMARY AND CONCLUSIONS

2.1. Presentation of the Active Substance

2.1.1. Identity, Physico-Chemical Properties & Methods of Analysis

The active substance margosa extract is an extract derived from ground seed kernels of the tropical neem tree Azadirachta indica using the manufacturing method developed by the applicant. Due to the complex nature of the botanical insecticide margosa extract, the main constituent - azadirachtin A (AzaA) - is used as the analytical lead compound.

Margosa extract is a plant extract and consists mainly of the limonoids azadirachtin A, azadirachtin B, azadirachtin H, Desacetyl-Nimbin, Desacetyl-Salannin, Nimbin, Salannin together with co-extracted fatty acids and a small amount of water and other non-accounted materials. Margosa extract may contain traces of aflatoxins.

Margosa extract is thermally stable up to 200 °C, where it begins to decompose. The vapour pressure should be << 10-5 Pa based on the calculated vapour pressure of 3.6·10-13 Pa for azadirachtin A. The water solubility and the partition coefficient can not be determined for extraction mixtures.

Margosa extract is neither highly flammable (no relative self-ignition up to the melting point), explosive nor has oxidising properties. In conclusion, no hazard indication is required for the active substance margosa extract.

Sufficiently validated analytical methods for the determination of azadirachtin A, the other limonids and impurities in the technical material are available. The determination of azadirachtin A is considered as the enforcement method, but methods for determination of all limonoids are available.

Identity, Physico-chemical Properties and Method of Analysis of the Insecticide NeemAzal-T/S

The biocidal insecticide NeemAzal-T/S is an emulsifiable concentrate containing further ingredients beside the active substance margosa extract. Due to the nature of the biocidal product it is not expected to exhibit any hazardous physical-chemical properties. It was noted that due to the limited thermal stability of the limonoids in margosa extract, the product may show limited shelf life, which can possibly be solved by recommending storage conditions (maximum temperature specified) and/or end use date.

The analytical methods for NeemAzal-T/S in all relevant matrices are covered by the methods of azadirachtin A. Due to the complex nature of margosa extract, the choice of azadirachtin A as lead compound for residues in soil and water is accepted.


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2.1.2. Intended Uses and Efficacy

The assessment of the biocidal activity of the active substance demonstrates that it has a sufficient level of efficacy against the target organism and the evaluation of the summary data provided in support of the efficacy of the accompanying product, establishes that the product may be expected to be efficacious.

In addition, in order to facilitate the work of Member States in granting or reviewing authorisations, and to apply adequately the provisions of Article 5(1) of Directive 98/8/EC and the common principles laid down in Annex VI of that Directive, the intended uses of the substance, as identified during the evaluation process, are listed in Appendix II.

The aim of the treatment is to combat Thaumetopoea processionea caterpillars in order to prevent the formation of stinging and allergenic hair in the surrounding air which evoke allergic reaction in human and animal. As the dead caterpillars and the exuviae also contain the stinging hairs, margosa extract would not be able to directly reduce the number of hairs in the air, because it only kills the caterpillars. Direct reduction of the stinging hairs is only possible by physical methods. Nevertheless, larvae develop the stinging hairs in high amounts beginning with the 3rd larval instar. Therefore, it is expected that the treatment will lead to a significant prevention of the formation of hairs in the air, when it is performed at a time when the predominant part of the caterpillars is in the 1st or 2nd larval instar. The efficacy is therefore determined by the killing effect of margosa extract, which in consequence leads to a prevention of the formation of stinging hairs in the air provided that the treatment is performed at an early larval stage (1st or 2nd larval instar). Therefore, the use of margosa extract has to be restricted to professionals, because special expertise is needed.

The effectiveness of Neem extracts against different Order of insects is confirmed in the literature. The extract and the product deter feeding in insects and disturb the neurosecretory organs, inducing malformations and consequently death. The specific studies provided in the dossier are quite simple in methodology but confirm the efficacy in the laboratory and in the field of the product against different instar stages including 2nd instar caterpillars of one specific target organism for which the product is claimed, i.e., caterpillars of T. processionea.

Thus, in a laboratory study NeemAzal T/S is effective at a concentration of 0.3 % (corresponding to 0.009% margosa extract) which corresponds to the claimed application rate. 10L of the 0.3% diluted product are used per tree.

Occurrence of Resistance

Margosa extract is a plant extract from seed kernels of the Neem tree. Due to the complex composition of this plant extract and the complex mode of action which is not restricted to a single target site, the possibility of the development of resistance against this active ingredient is estimated as very low.


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2.1.3. Classification and Labelling

Classification and Labelling of Margosa extract

Up to now there is no legal classification. Evaluation of the submitted data under Directive 98/8/EEC resulted in the following proposal for classification and labelling:

Table 2-1 Proposed classification of margosa extract

Remark: Classification according to the new Regulation (EC) 1272/2008 on classification, labelling and

packaging of substances and mixtures (CLP): H317 (May cause an allergic skin reaction); H361d (Suspected of

damaging fertility or the unborn child); H411 (Aquatic Chronic), category 2

In deviation to participant’s proposal for classification and labelling, margosa extract is classified as sensitising by skin contact in view of the results in a Magnusson & Kligman Test. The observed malformations in a rat teratogenicity study are considered relevant for a classification as toxic to reproduction Category 3.

The environmental classification is derived from the available acute tests with fish with the product NeemAzal-T/S. According to the findings of these tests, and taking into account also the results of the chronic fish test with margosa extract, there is evidence that the LC50 to fish of margosa extract is in the range of > 1 mg a.s./L and < 10 mg a.s./L. margosa extract is degraded in aqueous systems by both biotic and abiotic processes, however, it does not fulfill the ‘readily biodegradable’-criteria. With regard to the environmental classification, the safety phrases ‘S60, S61’ are mandatory.

The compound is neither highly flammable (no relative self-ignition up to the melting point), explosive nor has oxidising properties. In conclusion, no hazard indication is required for the physical and chemical properties for the active substance.

Classification Proposed

Class of danger Xi Irritant Xn, Repr. Cat. 3 Harmful N Dangerous to the environment

R 43 May cause sensitisation by skin contact

R 63 Possible risk of harm to the unborn child

R51 Toxic to aquatic organisms.

R phrases

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

(S 2) Keep out of the reach of children S 36/37 Wear suitable protective clothing

and gloves S60 This material and its container

must be disposed of as hazardous waste.

S phrases

S61 Avoid release to the environment. Refer to special instructions / material safety data sheet.


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Classification and Labelling of NeemAzal-T/S

Table 2-2 Proposed classification of NeemAzal-T/S

The classification of NeemAzal-T/S is done on the basis of study results presented in the dossier, according to the principles of Directive 67/548/EEC and Directive 1999/45/EC.

Besides the active substance margosa extract (content in the preparation: up to 4% w/w), the preparation contains formulation additives, which have not to be classified with regard to human health and environment. Margosa extract is classified as sensitising by skin contact (R 43) and Repr. Cat. 3 R 63. NeemAzal-T/S is not classified as sensitising based on the results of a maximisation test and it is not classified as R 63 because the concentration of margosa in NeemAzal is below the concentration limit of > 5 % in preparations.

Although margosa extract is identified as dangerous to the environment (R51, R53), its concentration in the preparation is below the limit (≥ 25% w/w) that triggers the classification of the preparation.

This evaluation is also supported by the available ecotoxicological data of the preparation showing an LC50 value for fish of > 100 mg/L.

Summary and conclusion:

Classification and labelling of the insecticide NeemAzal T/S is not required.

2.2. Summary of the Risk Assessment

2.2.1. Human Health Risk Assessment

2.2.1.1. Hazard identification

Absorption, Distribution, Excretion and Metabolism

No studies on absorption, distribution, metabolism and excretion were submitted. Margosa extract is a plant extract and contains many known but also unknown constituents. The active compounds of the neem kernels are not known, the triterpenoids (known as limonoids) and among them the azadirachtins are considered as the most relevant in effectiveness against insects. Azadirachtin A is treated as the lead compound of margosa extract but it is unknown,

Classification

according Directive

1999/45/EC

Proposed

Indication of danger Not required

R phrases None Not required

S phrases None Not required


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if this substance is also the most relevant with regard to toxicological aspects. Neither azadirachtin A nor any other limonoid is available as radioactive compound. Therefore, based on lack of technical feasibility, it is acceptable that no studies on metabolism and toxicokinetics were submitted.

No guideline studies on dermal absorption were submitted. The submitted study (isolated perfused bovine udder model) fulfils the general criteria for scientific acceptance, and a low degree of dermal absorption was observed. However, as the extent of absorption for individual components of the extract was unknown, it was agreed at the TM-III-10 to use a value of 100 % dermal absorption for further risk assessment.

Acute Toxicity

Margosa extract is not acutely toxic when administered orally, dermally or by inhalation. It is neither irritating to the skin nor to the eyes of rabbits. Margosa extract is a skin sensitiser.

Classification/labelling for sensitisation according to Directive 67/548/EEC:

Xi; R43

Classification/labelling for sensitisation according to GHS, (EC) 1272/2008:

H317 (May cause an allergic skin reaction)

Short-term Toxicity

In a 28-d rat feeding study with margosa extract, the animals of all dosed groups showed histological changes in the liver (eosinophilia, hepatocyte hypertrophy) increased liver weight and, additionally, thyroid follicular epithelial hypertrophy. Therefore, a NOAEL could not be identified, the lowest dose tested was 3200 ppm (300 mg/kg bw/d).

In a 90-d rat feeding study with margosa extract, the top dose of 6400 ppm (490 mg/kg bw/d) induced hypertrophy of hepatocytes and thyroid follicular epithelial cells as well as an increase in liver and thyroid weight. Additionally, some changes in haematological parameters (prolonged APTT in males) were observed. At 1600 ppm (123 mg/kg bw/d), slight changes in a few clinical chemistry parameters and histological changes in liver histology (periportal fat deposition) in females were found. The NOAEL was 400 ppm (32 mg/kg bw/d).

An appropriate repeated dose toxicity study with margosa extract in a second (non-rodent) species is lacking. The applicant justified non-submission by presenting published reports on the administration of neem seed products (water-washed neem seed kernel cake) to farm animals where no toxic effects were observed. Although the data obtained from these publications are not suitable to replace standard toxicity studies and uncertainties remain, it may be concluded that there was no indication for a high sensitivity or different target organs in non-rodents following (sub)chronic exposure. Therefore, non-submission was accepted.


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Genotoxicity

In vitro and in vivo tests provided no evidence for a genotoxic potential of margosa extract.

Chronic Toxicity/ Carcinogenicity

No carcinogenic potential and no adverse effects were found in the rat long-term toxicity study for the margosa extract, leading to a NOAEL of 6400 ppm (equivalent to 448 mg/kg bw/d).

The mouse carcinogenicity study was conducted with the NeemAzal-F5 % formulation instead of margosa extract. This study presents several limitations and is therefore not fully acceptable. However, macroscopic and histopathological examinations did not reveal any substance-related increase in the occurrence of neoplastic lesions up to the highest dose tested (i.e. ca. 12.6 mg margosa extract/kg bw/d).

Based on the absence of any carcinogenic potential up to the highest dose levels in the long-term studies in mice and rats, the absence of any other treatment-related adverse effects in the long-term rat study as well as in subchronic studies in a variety of other mammalian species, on the absence of a genotoxic potential in in vitro and in vivo genotoxicity tests, on the current discussion of the usefulness of the species mouse in long-term carcinogenicity studies and, finally, on animal welfare considerations no further long-term study in a second species is required.

Reproduction Toxicity

The developmental toxicity of margosa extract has been evaluated in the rat (Trifolio GmbH, here referred to as: NeemAzal technical) and in the rabbit performed with a different margosa extract. The results of the study evaluating the developmental toxicity in rabbit are read across to evaluate NeemAzal technical based on the similar toxicological profile of the extracts in subchronic rat studies.

In the rat teratogenicity study with NeemAzal technical, an increased incidence of heart malformations in offspring were observed at 225 mg/kg bw/d and above. At 1000 mg/kg bw/d, maternal toxicity was evidenced by decreased body weight gain. The NOAEL for maternal toxicity was therefore 225 mg/kg bw/d. The NOAEL for teratogenicity and embryotoxicity was 50 mg/kg bw/d.

In the rabbit teratogenicity study, the top dose of 500 mg/kg bw/d induced an increase in abortions and resorptions, and the remaining few live foetuses showed an increased incidence of malformations. At the same dose level, dams showed severe body weight loss from gestation days 6 to 12, resulting in a net body weight loss at study termination. At 100 mg/kg bw/d, body weight of dams was also slightly reduced between gestation days 6 and 12, but subsequently body weight gain increased, resulting in a net body weight gain. At 100 mg/kg bw/d, no effect on offspring was observed. The NOAEL for maternal toxicity was 20 mg/kg bw/d. The NOAEL for teratogenicity and embryotoxicity was 100 mg/kg bw/d. Malformations were observed in the high-dose group only and may be related to maternal toxicity.


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Developmental toxicity was observed in both studies (rat and rabbit). In the rat study, specific malformations (affecting the heart) were found in the mid- and high-dose groups and an increased incidence of supernumerary rib was noted in the high-dose group. However, the heart malformations were also seen in the mid-dose group without clear adverse effects in the mothers; therefore they are not obviously produced by a non-substance-specific secondary mechanism.

In a rat two-generation study with margosa extract, up to the highest dose tested of 750 ppm (51 mg/kg bw/d) treatment-related effects were observed neither in parental animals nor with respect to reproductive parameters. Therefore, the NOAEL was the highest dose tested of 750 ppm (51 mg/kg bw/d).

In another supplementary (serious reporting deficiencies) rat two-generation study with NeemAzal-F 5 % (containing 5 % azadirachtin), increased relative testis and spleen weight was observed in all treated groups of the P0.generation. Additionally, body weight gain in the mid- and high-dose groups was reduced. No treatment-related effects on reproductive performance were observed.

Classification/labelling for reproduction toxicity according to Directive 67/548/EEC:

Toxic to reproduction category 3, substances which cause concern for humans owing to possible developmental toxic effects, Xn R 63: Possible risk of harm to the unborn child.

Classification/labelling for reproduction toxicity according to GHS, (EC) 1272/2008:

H361d (Suspected of damaging fertility or the unborn child)

Neurotoxicity

In a non-GLP study, three groups of six female chickens each were dosed daily by gavage with suspensions of margosa extract at dose levels of 0, 500, 750, and 1000 mg/kg bw/d for 21 days. The control group received distilled water. On day 22, 50 % of the birds were sacrificed and the remaining birds were observed for another 21 days. All birds were sacrificed on day 43. The following parameters of neurotoxicological relevance were investigated: a daily behavioural test for locomotive ataxia, activity of acetylcholine esterase in blood and serum on day 0, 23, and 43, histopathological examination of the brain (cerebrum, cerebellum, medulla oblongata), spinal cord (thoracic, cervical, lumbo-sacral) and sciatic nerve (proximal to distal length on either side) collected following sacrifice. In addition, animals were observed for clinical signs of toxicity, body weight and food consumption as well as number and weight of eggs laid. Blood was analysed for haematological and biochemical parameters. No treatment-induced effects were observed regarding neurotoxicity parameters or general toxicity. No treatment-related changes were noted up to the highest dose group and a NOEL of 1000 mg/kg bw/d was established in this study.


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As margosa extract is neither an organophosphorous compound nor was the nervous system a target in the repeated dose toxicity studies, no further study is required.

Mechanistic Studies

No studies have been submitted. Waiving is justified considering that no specific effects are reported which would require a mechanistic clarification for further risk assessment.

Medical Data

Evaluation of the literature on neem demonstrates evidence of poisoning incidents and side-effects in the use of neem products. "Margosa Oil" or "Neem Oil" is used as a traditional medicine in Asia and Africa. Case reports describe severe intoxications in children. Vomiting, drowsiness, convulsions, metabolic acidosis, and encephalopathy are among the reported signs of poisoning, autopsy of fatal cases revealed liver damage. According to some authors, the findings resemble those of Reye's syndrome.

It is not possible to conclude on the toxic principles underlying the reported cases. According to some reports, aflatoxin residues may explain some poisoning incidents with the oil, while other authors assume an inherent toxicity of neem oil constituents.

Anti-fertility (contraceptive and abortive) effects of oils and extracts are reported in studies with various mammalian species including humans (overview e.g. Schmutterer H., 2002, The Neem Tree, Mumbai).

All clinical cases are reported for neem-derived materials, specifically for leaves and oil, but not for the margosa extract of the applicant (NeemAzal technical). Medical observations of workers in the production of margosa extract did not show adverse health effects in the three years observation period.

Biocidal Product

NeemAzal-T/S exhibited low acute oral, dermal and inhalative toxicity to male and female rats. The product NeemAzal-T/S is considered to be non- irritating to the skin and eye. Two studies for evaluation of skin sensitising potential (Magnusson-Kligman-Test, Buehler Test) lead to the conclusion that NeemAzal-T/S is considered as not sensitising to skin.

No classification and labelling of the product NeemAzal-T/S is required.


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2.2.1.2. Effects assessment

Summarising the study results and all considerations above, margosa extract requires classification/labelling according to Directive 67/548/EEC as follows:

� Sensitisation by skin contact, Xi, R 43

� Developmental toxicity Cat. 3, Xn, R 63

Based on malformations observed the teratogenicity study in rats, the NOAEL of 50 mg/kg bw/d (LOAEL = 225 mg/kg bw/d) is regarded as the relevant starting point for setting a systemic reference dose for acute exposure.

By applying a default assessment factor of 100, a systemic acute Acceptable Exposure Level (AELacute) of

0.5 mg/kg bw/d

is proposed for acute exposure to margosa extract.

The NOAEL of 32 mg/kg bw/d from the 90-d toxicity studies in rats (LOAEL = 132 mg/kg bw/d) is regarded as the relevant starting point for setting a systemic reference dose for medium- and long-term exposure.

By applying a default assessment factor of 100, a systemic medium-/long-term Acceptable

Exposure Level (AELmedium-/long-term) of

0.32 mg/kg bw/d

is proposed for medium-/long-term exposure to margosa extract.

2.2.1.3. Exposure assessment

Exposure of Professionals

The active substance margosa extract is produced outside the EU. The biocidal product NeemAzal T/S (3 % a.s.) is produced within the EU. The occupational exposure is assessed for the formulation of the biocidal product. The results are not further evaluated for risk characterisation purposes under the requirements of the BPD.

The following scenarios are covered by the exposure assessment in this report:

• Occupational exposure from use of biocidal product (spraying)


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• Secondary professional exposure

The biocidal product is applied to control the oak processionary moth, Thaumetopoea

processionea, first and second caterpillar stage. The predominant aim of control of this pest is to avoid contact to the hairs of oak processionary moth caterpillars during outdoor activities. Therefore, oak trees located in amenity areas and bearing these caterpillars have to be treated. The product will be applied with a hand-held spraying device from a lifting platform on crown level of the treated trees. Users are e.g. landscape conservation units, forest service, fire brigades. Before spraying the biocidal product NeemAzal T/S (3 % a.s.) is diluted to a spraying solution of 0.009 % active substance.

No information is supplied by the participant how many days an individual is exposed to the biocidal product. The rapporteur uses the following worst case assumption: For the personnel of landscape conservation units of communities application is restricted to 6 weeks or at most 36 days per year.

In the case of specialized companies the period of application could be extended up to 8 weeks or up to 48 days per year.

Two calculation models, the German BBA and the UK POEM were applied to estimate operator exposure. Due to the low vapour pressure of the active substance inhalation exposure during mixing & loading and post-application phase of the biocidal product was excluded from the exposure calculation. Moreover the formation of aerosol is not expected during these phases. The resulting level of exposure calculated by German BBA and UK POEM model are in the same order of magnitude. For risk assessment purposes the result of the German BBA model is taken forward. The resulting inhaled amount is 0.014 mg a.s./person/day.

Dermal exposure is expected during all phases of exposure. The assessment of the dermal exposure is based on the German BBA model for the mixing & loading and application phase. The dermal exposure during the post-application phase is assessed by the Riskofderm model. The resulting level of potential dermal exposure is 10.7 mg a.s./person/day.

Exposure of bystanding professionals (secondary exposure) will be lower as compared to operator exposure. Since operator exposure was estimated to be well below the AEL even in the absence of PPE, no calculations or measurements are necessary to conclude that exposure of bystanding professionals will be below exposure limits.

For further details please see Appendix I - List of endpoints - Acceptable exposure scenarios.

Exposure of Non-Professionals

Primary Non-Professional Exposure

Primary exposure to non-professionals is not intended since the biocidal product in this dossier is foreseen for professional use only. However, it is likely that biocidal products for


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non-professional use others than those against the caterpillars of the oak processionary moth will be submitted for national authorisation according to directive 98/8/EC. Therefore, a realistic worst-case-exposure assessment for non-professionals for the use in home and garden according to the German model was performed, resulting in a systemic exposure estimate of 0.0732 mg/kg bw/d. Medium-term is assumed since the non-professional user may apply the biocidal product frequently over the summer season. Thus, this is equivalent to 22.9% of the AELmedium-term (0.32 mg/kg bw/d). Thus, exposure of non-professionals using a biocidal product equivalent to NeemAzal-T/S would be acceptable in relation to human health.

Secondary exposure to margosa-extract of the general public (consumer)

Secondary exposure of the general public / bystanders is expected during professional application of the biocidal product. After application dermal exposure to the biocidal product is possible if persons re-enter and stay for a longer time period in areas, in which trees have been treated. Chronic secondary exposure is not expected for the general public.

Table 2-3 Summary of secondary internal exposure of the general public (consumer) to

margosa extract from NeemAzal-T/S

Dermal exposure

(mg/kg bw[/d])

Inhalation exposure

(mg/kg bw[/d])

Modeled data

Acute/Medium-term exposure – internal

dose

Adults, bystanders 0.1460 0.00001

Adults, re-entry, residents 0.00533 not relevant

Infants, bystanders 0.2628 0.00004

Infants, re-entry, residents 0.01139 not relevant

Exposure via Residues in Food

No residues in food are expected with the intended use.

Combined Exposure

Margosa extract is used at most a few times during a period of some weeks per year. Exposure of the general public can be neglected; therefore no concern is anticipated for a professional and non-professional combined exposure. However, margosa extract is used also as a pesticide in agriculture. It is not probable that the same persons are involved in the use of margosa extract both as a biocidal product and as a plant protection product in agriculture. However, this possibility cannot be excluded and therefore, as a worst case, it can be assumed


18

that the same persons would be involved in agricultural and biocidal use of margosa extract. The frequency of these combined exposures is not expected to go beyond medium-term time-frames; therefore a potentially combined agricultural and biocidal use of margosa extract is also adequately addressed in the risk characterisation of professional use in this report.

2.2.1.4. Risk characterisation

Risk Assessment for Professionals

In a first step (Tier 1) occupational risk assessment is based on the internal reference dose (AEL medium-/long-term) of 0.32 mg/kg/day. A comparison with potential exposure gives a rough but cautious assessment to decide on concern (table 2-3). Under the specific conditions described there is no concern for the scenarios evaluated in this report.


19

Table 2-4 Risk characterisation for professionals using Tier 1

potential exposure

(external values) Exposure scenario Specific conditions

inhalation

mg/kg bw/d

dermal

mg/kg

bw/d

Total potential

internal dose

(mg/kg bw/day)

(1)

Total

internal

body

burden

divided by

AEL (2)

Application of the biocidal product

Mixing and loading

45 mL NeemAzal T/S (3 % a.s.) is diluted with water in batches of 15 L - final concentration is 0.009 % a.s.

Form of exposure: liquid (3 % a.s.)

Duration: 5-10 min

0.124 0.124 0.3875

Spray application

Spraying of 0.009 % a.s., 10 L per tree

Form of exposure: aerosol (0.009 % a.s.)

Duration: 30 – 60 min. per task

Frequency: Approximately 10 - 20 trees may be treated per day, 8 weeks or at most 48 days per year

0.00023 0.055 0.055 0.172

Post-Application

Reloading of the spraying device, cleaning of spraying device

Form of exposure: liquid (0.009% a.s.)

Duration: 30 – 60 min.

0.0008 0.0008 0.0025

Total 0.18 0.56 (1)

100% systemic availability after inhalation and dermal exposure (2)

AEL medium-/ long-term = 0.32 mg/kg/day

Summary and conclusion:

The following table summarise the risk characterisation outcome for professionals.


20

Table 2-5 Professional Users – Primary Exposure (margosa)

Estimated Internal Exposure

Exposure Scenario

(indicate duration)

estimated

oral

uptake

[mg/kg

b.w/day]

estimated

inhalation

uptake

[mg/kg

b.w/day]

estimated

dermal

uptake

[mg/kg

b.w/day]

estimated

total uptake

[mg/kg

b.w/day]

Relevant

NOAEL/

LOAEL

[mg/kg

b.w/day]

&

Reference

Value

e.g: AEL

(acute or

medium or

chronic)

AF

MOEre

f

MOE Exposure

/AEL

Mixing and loading 5-10 min./task max. 48 days /year

0.124 0.124 258 0.3875

Spray application 30-60 min/task max. 48 days/year

0.00023 0.055 0.055 582 0.172

Post-application 30-60 min./task max. 48 days/year

0.0008 0.0008 40000 0.0025

Tier 1

(no PPE)

Total 0.00023 0.1798 0.18

NOAEL=32 mg/kg/b.w./d

AEL medium-

/longterm=0.32 mg/kg b.w./d

100

178 0.56

Tier 2

(Refinement, PPE or other risk mitigation measures – Specify)

Tier 2 is not required

The occupational risk assessment for margosa extract is based upon the AEL approach and the estimate of potential occupational exposure. The risk assessment is considered to be sufficiently comprehensive and reliable for the purposes of Annex I inclusion of margosa extract.

For the application of the biocidal product the risk assessment does not lead to concern. It is essential to recognize that this conclusion only applies to the active substance (margosa extract) in the biocidal product. From the point of view of occupational safety and health there is no risk-related reason for conditioning the requested Annex I inclusion for margosa extract.


21

However, against the background of experimental evidence of developmental toxicity, it needs to be stressed, that the AEL for margosa extract should be adhered to on every single day of an exposure period.

Risk Assessment for Non-Professionals

Primary exposure

Since primary exposure to non-professionals is not expected a risk characterisation is not required. However, a realistic worst-case-exposure assessment for non-professionals for the use in home and garden according to the German model was performed, resulting in acceptable exposure estimates (table 2-5).

Table 2-6 Non-Professional User – Primary Exposure (margosa)


Exposure Scenario

(indicate duration)

estimated oral

uptake

[mg/kg

b.w/day]

estimated inhalation

uptake

[mg/kg

b.w/day]

estimated dermal uptake [mg/kg

b.w/day]

estimated total uptake

[mg/kg

b.w/day]

Relevant NOAEL/

LOAEL

[mg/kg

b.w/day]

&

Reference Value

e.g: AEL (acute or

medium or chronic)

AF

MOEref

MOE Exposure

/AEL

Tier 1

(no PPE)

Non-professional use, adult, Home & Gar-den, mixing & loading and application, repeated use, medium-term exposure

- 1.15 x 10-4 7.32 x 10-2 7.32 x 10-2 NOAELmedium

-term: 32

AELmedium-

term: 0.32

100 437 0.23

Tier 2

(Refinement or other risk mitigation measures – Specify)

Tier 2 is not required


22

Secondary exposure

For risk characterisation the systemic AELmedium-/long-term has been applied. This AEL derives from a 90-d rat study, which results in a NOAEL of 32 mg/kg/d. Applying an assessment factor of 100 leads to a systemic AELmedium-term of 0.32mg/kg bw/d.

Although repeated exposure is unlikely the AELmedium-/long-term has been used for risk characterisation instead of the AELacute representing worst case conditions. Additionally, the use of the AELacute, which was derived from a developmental study, is not applicable for infants. In Tier 1 risk assessment the exposure estimates are below the AELmedium-term. Thus it is concluded that secondary exposure of non-professionals by the biocidal product NeemAzal-T/S is acceptable in relation to human health.

Table 2-7 Summary – Tier 1 risk assessment for secondary exposure


Exposure Scenario

(indicate duration) estimated

oral uptake

[mg/kg

b.w./day]

estimated inhalation

uptake

[mg/kg

b.w./day]

estimated dermal uptake [mg/kg

b.w./day]

estimated total uptake

[mg/kg

b.w./day]

Relevant NOAEL/

LOAEL

[mg/kg

b.w./day]

&

Reference Value

e.g.: AEL (acute or

medium- or long.-term)

AF

MOEref

MOE Exposure

/AEL

Tie

r 1

(W

ors

t C

ase

)

Med

ium

-ter

m S

cen

ari

o

Bystander, adult, repeated exposure medium-term exposure

- 1.46 x 10-1 1.00 x 10-5 1.46 x 10-1 NOAELme-

dium-term: 32

AELmedium-

term: 0.32

100 219 0.46


23

Bystander, infant, repeated exposure, medium-term exposure

- 2.63 x 10-2 4.00 x 10-4 2.63 x 10-1 NOAELme-

dium-term: 32

AELmedium-

term: 0.32

100 122 0.82

Resident, re-entry, adult, repeated exposure medium-term exposure

- 5.53 x 10-3 - 5.53 x 10-3 NOAELme-

dium-term: 32

AELmedium-

term: 0.32

100 6004 0.02

Resident, re-entry, infant, repeated exposure, medium-term exposure

- 1.14 x 10-2 - 1.14 x 10-2 NOAELme-

dium-term: 32

AELmedium-

term: 0.32

100 2809 0.04


24

Table 2-8 Summary - risk assessment for secondary exposure

Exposure scenario Exposure

(mg/kg

bw/d)

AELmedium-term

(mg/kg bw/d)

Exposure

(% of AELmedium-term)

MOE

Acute/Medium-term exposure –

internal dose

Adults, bystanders, dermal, inhalation

0.1460 0.32 46 219

Adults, re-entry, residents, dermal 0.00533 0.32 1.7 6004

Adults, total 0.1513 0.32 47 212

Infants, bystanders, dermal, inhalation

0.2628 0.32 82 122

Infants, re-entry, residents, dermal 0.01139 0.32 3.6 2809

Infants, total 0.2742 0.32 86 117

The exposure estimates are below the AELmedium-term. Thus it is concluded that secondary exposure of non-professionals by the biocidal product NeemAzal-T/S is acceptable in relation to human health.

2.2.2. Environmental Risk Assessment

2.2.2.1. Fate and distribution in the environment

The use of a non-labelled test substance and the non-feasibility of synthesising reference compounds result in an unavoidable limitation of the findings concerning the fate and behaviour of margosa extract i.e., the impossibility to determine the pathway of degradation of the test substance and the quality and quantity of its degradation products in a number of environmental fate studies (e.g., soil and water/sediment degradation, hydrolysis, aqueous photolysis).

The findings for Aza A are considered in general to be representative at least for the other azadirachtins, i.e., Aza B and Aza H, of margosa extract, and therefore for a total portion of more than 40 % by weight of the active substance margosa extract.

Biodegradation

A total of five ready biodegradability tests were conducted: one with the formulated product NeemAzal-T/S, three with the active substance margosa extract (34 % azadirachtin A) and one with azadirachtin A. Only the product NeemAzal-T/S (containing 1 % azadirachtin A)


25

fulfilled the criteria of classification as ‘readily biodegradable’ (i.e., > 60 % degradation within 10 days), probably attributable to the properties of the formulation additives, whilst neither margosa extract nor azadirachtin A proved to be readily biodegradable.

No standard water/sediment study is available. Only literature data for the compound azadirachtin A were presented, which can only be regarded as additional information and will not be used for PEC estimation. Since radiolabelling of the main components of margosa extract was technically not feasible, the determination of bound residues, mineralisation rates and relevant metabolites was not possible. The extremely complex structure of azadirachtins also hampers the elucidation of metabolic and degradative pathways by exclusively analytical methods like HPLC, TLC, GC or spectroscopic methods.

The dissipation rates of azadirachtin A in the different water systems are rapid to delayed. It can be assumed that hydrolysis is the main degradation process.

The half-lives from literature data for azadirachtin A vary from 2.5 to 66 days converted to an average EU outdoor temperature of 12 °C.

Summarising the findings of three aerobic laboratory soil studies the DT50 values for azadirachtin A varies between 1.9 and 37.9 days in six soils at 20 °C resulting in a geometric mean of 5.45 days. Converted to an average EU outdoor temperature of 12 ºC the half-lives range between 3.6 and 71.9 days, the geometric mean is 10.34 days (n=6). The DT50 values of azadirachtin B amount to 39.7 and 99.0 days in two different soil types converted to 12 ºC. Since the tests were conducted with non-labelled material, neither mineralisation nor the formations of bound residues were quantifiable.

In general quantification of the intermediate fractions and mass balance, however, were not possible with regard to the technical limitations linked to the work with unlabelled material. The study results conducted with unlabelled test substance show that azadirachtins and proposed metabolites indicate a rapid to delayed primary degradation.

Abiotic Degradation

The hydrolytic stability of azadirachtin A is strongly pH-dependent as indicated by a significant increase in the rate of degradation with increasing pH. The extrapolation of the test results to a temperature of 12 °C using the Arrhenius equation yields a half-life of 117.7, 40.9, and 8.2 days at pH 4, 7, and 8, respectively. Hydrolysis products were not identified due to the technical limitations with regard to the impossibility of radiolabelling of the test substance and synthesis of reference substances.

Aqueous photolytic half-lives for margosa extract were calculated based on the quantum yield and UV/VIS data from the direct phototransformation study in water of margosa extract and


26

parameters included in the computer model “ABIWAS”. The average half-life was estimated as 1.8 months for January and 5.5 days for July.

With regard to the estimated half-life of 1.696 h (equivalent to 0.071 d) for azadirachtin A, long-term transport and accumulation in air are not to be expected. Furthermore, the tendency of azadirachtins, the major components of margosa extract, to enter the atmosphere is considered to be low taking into account both the vapour pressure of these compounds (3.6 x 10-13 Pa) and the Henry’s Law Constant (2.4 x 10-14 Pa m3/mol).

Distribution

Results from two adsorption/desorption studies with azadirachtin A are available and give a median Koc value of 75.2 mL/g. Due to the low Koc value in the tested soils, azadirachtin A is slightly adsorbable to soil.

Mobility

The high mobility of azadirachtin A in soil as indicated by the low KOC is confirmed under the stringent conditions of the laboratory column leaching test, i.e., highly exaggerated concentration of substance applied to soil, maximum water saturation of soils at test start, watering with 200 mm rain within two days following test substance application.

However, contamination of groundwater by azadirachtin A under actual use conditions seems to be unlikely taking into account its short degradation half-life in soil (see above).

Bioaccumulation

Bioconcentration factors for the aquatic (BCFfish = 2.5) and the terrestrial compartment (BCFearthworm = 1.1) were estimated on basis of the highest log Pow of 1.29 for azadirachtin B. The log Pow for azadirachtin A is 0.99 and the log Pow for azadirachtin H is analysed as 0.68. The calculated BCF values indicate that a main component of margosa extract has a low potential to bioaccumulate in aquatic and terrestrial organisms and would pose no unacceptable risk of biomagnification in the food chain of either compartment.

2.2.2.2. Effects assessment

Ecotoxicity tests both with the active substance margosa extract as well as the biocidal product NeemAzal-T/S are available. Tests with the product are considered as adequate for the effects assessment of the active substance as according to the available data on the formulation additives, the ecotoxicity of the b.p. is expected to be associated with the a.s. rather than any of those additives.


27

Aquatic Compartment

The database of acceptable laboratory tests that is available for margosa extract and the product NeemAzal-T/S comprises both acute and long-term toxicity tests with fish, invertebrates and algae. The lowest effect value from the acute studies was obtained for fish (96h-LC50 = 4.14 mg a.s./L). However, the effect value for Daphnia was only a factor of 2 higher (48h-EC50 = 9.69 mg a.s./L). Algae are by orders less sensitive (72h-ErC50 = 1041 mg a.s./L; 72h-ErC10 = 332 mg a.s./L). In long-term tests performed with the product NeemAzal-T/S Chironomus riparius was most sensitive (28d-NOEC = 0.006 mg a.s/L). For Daphina a 21d-NOEC of 0.1 mg a.s./L was derived while for fish the 28d-NOEC is 1.9 mg a.s./L.

With the available data a PNECwater for margosa extract of 0.00006 mg a.s./L (0.6 µg a.s./L) was derived by applying an assessment factor of 10 to the lowest NOEC, as long-term tests with species from three trophic levels are available.

Sediment

PNECsediment was derived from the PNECwater using the equilibrium partitioning method according to the TGD resulting in a PNECsediment of 0.00145 mg a.s./kg ww (1.45 µg a.s./kg ww).

Inhibition of microbial activity

In a standard activated sludge respiration inhibition test with sludge from domestic sewage treatment plant a NOEC of 1000 mg a.s./L and an EC50 of > 1000 mg a.s./L were determined. The risk to the micro-organism population of a sewage treatment plant can be characterised to be low to negligible regarding the results of these studies. From this data a PNECmicro-organisms

of 100 mg a.s./L (nominal) was derived.

Atmosphere

For azadirachtins, i.e. the major components considered also responsible for the biological activity of margosa extract, physico-chemical properties have been determined resulting in low vapour pressure (3.6 x 10-13 Pa) and the Henry’s Law Constant (2.4 x 10-14 Pa m3/mol). Thus, negligible volatilisation and transfer to air of azadirachtins are to be expected. Additionally, the chemical half-life of azadirachtin A in the troposphere was calculated to be 1.7 hours. According to these findings, accumulation and long-distance transport of margosa extract in the air followed by wet or dry deposition is not to be expected.

Terrestrial Compartment

All terrestrial tests were conducted with the product NeemAzal-T/S and the effect values are recalculated to the content of margosa extract. Acute tests with earthworms (14d-EC50 = 12 mg a.s./kg dw) and plants (22d-EC50 > 0.06 mg a.s./kg dw), a reproduction test with a


28

predatory mite (NOEC = 5974 mg a.s./kg dw) as well as a soil micro-organism study (28d-NOEC = 1.28 mg a.s./kg dw) are available.

The PNEC for the terrestrial compartment is based on the long-term effect value for soil microorganisms (28d-NOEC = 1.28 mg a.s./kg dw). A PNECsoil of 0.02 mg a.s./kg ww (normalized to standard soil) was derived using an assessment factor of 100.

Margosa extract is slightly toxic to honey bees by oral and contact exposure (72h-LD50 = >17.7 and 63 µg a.s./bee). Furthermore semi-field test with the product NeemAzal-T/S sprayed on foraging plants indicate no unacceptable risk to honey bee brood at use rates up to 6 L product/L (176 g a.s./ha).

Several (laboratory, extended laboratory, semi-field and field) tests are available, evaluating the toxicity of NeemAzal-T/S to non-target arthropods.

Low toxicity of margosa extract to birds was found in an acute oral (14d-LD50 > 4000 mg a.s./kg bw) and a short-term dietary test (5d-LD50 > 5200 mg a.s./kg food). In a reproduction study a NOEC of 350 mg a.s./kg food was found for margosa extract. It has to be emphasized, that the applicant has no access to this study. Therefore, for product authorisation, a letter of access to this study needs to be provided.

Secondary Poisoning

Although margosa extract has only a low bioaccumulation potential, insectivorous birds and small mammals may be exposed to the substance when feeding on treated caterpillars in and beneath newly sprayed oak canopies. Therefore, an assessment of secondary poisoning for birds and mammals needs to be performed. A PNECoral,bird of 11.7 mg a.s./kg food corresponding to 0.82 mg a.s./kg bw/d was derived from the reproduction study using an assessment factor of 30. For mammals the PNECoral was derived from a rat developmental toxicity study using an assessment factor of 30, resulting in a PNECoral,mammal of 25 mg a.s./kg food corresponding to 1.7 mg a.s./kg bw/d.

2.2.2.3. PBT assessment

P-/vP-Criterion

Only literature data for the compound azadirachtin A in freshwater systems are available. Several outdoor studies indicate that azadirachtin A disappears rapidly from the water phase of natural waters with the highest dissipation rate of approximately 35 days at 20 °C (corresponding to 66 days at an average EU outdoor temperature of 12 °C).

Results of three degradation studies in soil indicate a rapid primary degradation of azadirachtin A and low persistence in soil with DT50 values ranging from 1.9 to 37.9 days at 20 °C (geo mean = 5.45 days). Converted to 12 °C the half-lives vary between 3.6 and 71.9 days at 12 °C (geo mean = 10.34 days). DT50 values for azadirachtin B taken from PPP regulation data amount to 20.9 and 52.2 days at 20 °C as well as 39.7 and 99.0 days at 12 °C.


29

The persistence criterion is not fulfilled for azadirachtin A and azadirachtin B.

B-/vB-Criterion

For azadirachtin A the calculated BCF in fish is 1.38 and 1.0 in earthworms, for azadirachtin B the BCFfish is 2.5 and BCFearthworm mounts up to 1.1. Therefore, neither the B- nor the vB-criterion are fulfilled for azadirachtin A and azadirachtin B, respectively.

T-Criterion

From a long-term test with Chironomus riparius performed with NeemAzal-T/S a NOEC value of 0.006 mg a.s./L related to margosa extract was derived. Therefore, the T-criterion is fulfilled.

Endocrine disrupting effects: Margosa seems to suppress hemolymph ecdysteroid and juvenil hormone titers on the neuroendocrine level. In the open literature, there are several papers dealing with endocrine effects of azadirachtin on various insect species. In aquatic studies with Daphnia magna a rather high acute-chronic-ratio of about 100 was found. In the long-term study with the benthic insect Chironomus riparius with emergence as endpoint, the NOEC of 0.006 mg/L indicates a specific effect on insect development that could be explained by the above mentioned effect.

No information is available on endocrine disrupting effects on further non-target organisms. Studies available in the human health risk assessment for margosa extract did not mention any indications for endocrine disrupting effects.

According to these considerations the main compound azadirachtin A of the a.s. margosa extract cannot be regarded as a PBT substance. Therefore, margosa extract itself is very likely no PBT substance.

2.2.2.4. Exposure assessment

The environmental exposures are assessed applying the EU Technical Guidance Document (TGD) on Risk Assessment (2003) and the OECD Emission Scenario Document (ESD) No. 18 for insecticides, acaricides and products to control other arthropods (PT18) for household and professional uses (2008) as well as the results of the “EU Workshop on Emission Scenario Documents for PT 18” held in Brussels (December 2007). For the environmental exposure assessment of direct release to surface water calculations according to FOCUS surface water were conducted.

The applicant has provided the data for one biocidal product (b.p.) NeemAzal-T/S which is only used by professionals for spraying onto infested solitary oaks. Emission scenarios for spraying in urban as well as in rural areas were taken into account.

For the environmental exposure assessment of margosa extract, the following life cycle stages are considered to be relevant:

� formulation of the b.p.,


30

� professional application of the b.p. (mixing/loading step, outdoor spray application as

well as wash-off from trees caused by rainfall),

All further life cycle stages (e.g. production of the a.s., release from private use, etc.) are not considered relevant in the present evaluation.

An environmental release estimation followed by an environmental exposure assessment is not performed for the life cycle stage “production” as margosa extract is produced outside the EU.

The formulation of NeemAzal-T/S takes place once a year in a manufacturing plant within the EU (in Germany). According to the applicant, the process occurs in a dedicated system which, following addition of the a.s. to the formulation mixture, continues in a closed system until the product is packaged and sealed into plastic containers. Waste cleaning fluids used during cleaning of equipment and process wastes are disposed of as dangerous wastes. These waste products are therefore not considered to be of concern for possible environmental exposures.

Furthermore, no emission to air and no accumulation of margosa extract in the atmosphere are to be expected due to the low vapour pressure of the major compounds of the a.s. and due to the formulation process not generating aerosols. In the national product authorisations it is recommended to re-check the formulation procedure of the product in a case-by-case manner whether the conditions of use described above are still maintained.

NeemAzal-T/S is intended for professional use, up to two applications per year with approx. two weeks between the applications. The product is used outdoors on oak trees to control caterpillars (1st or 2nd larval instar) of the oak processionary moth, Thaumetopoea

processionea. In the OECD ESD No. 18 (2008), emission scenarios for outdoor applications are developed for rural as well as urban areas which are both relevant for professional use of the b.p. NeemAzal-T/S.

Soil, groundwater, surface water and air compartment are at risk of direct exposure from the use of b.p. in rural areas, and STP and air compartment are directly affected from use in urban areas.

Mixing and loading as well as spray application including deposition and run-off during application together with wash-off from trees due to rainfall are addressed in emission calculations for urban as well as for rural environment. As a conservative approach, no degradation of the a.s. was taken into account as degradation studies were submitted for the main component azadirachtin A only.

The emission scenario “spray application (rural/urban)” was adapted to be in line with the specific biocidal application of the a.s. It is based on the emission scenario for flying insects; treated area and affected soil volume are adjusted.

The estimation of the local PECs for the aquatic compartment includes sewage treatment plant (STP), surface water, and sediment for the urban scenarios. For urban areas, 100 % of the releases end up to sewers. For the rural scenarios, direct release to the aquatic compartment is to be expected via spray drift.


31

The estimation of the local PECs for the terrestrial compartment includes soil and groundwater for the rural scenarios and groundwater for the urban scenario (exposure to soil via sewage sludge application).

For rural areas, releases will end up in soil (spray drift, run-off, wash-off) and surface water (spray drift). For the urban scenarios, release to paved ground will cause exposures of STP and the subsequent application of sewage sludge will lead to a certain exposure to soil. However, direct release to soil (considered in the rural scenario) will represent the worst-case for PECsoil, and additional exposure via sewage sludge underneath oak trees is not considered to be realistic. Therefore, PECsoil_sewage sludge is not calculated. PECgroundwater is calculated for both, rural and urban scenario.

No estimation of local PECs for the atmospheric compartment is performed as no enduring release to air is assumed because sprayed droplets are supposed to settle to the ground rapidly due to their size.

Insectivorous birds and small mammals may be exposed to the substance when eating treated caterpillars, and other insects, in and beneath newly sprayed oak canopies. Therefore, estimated theoretical exposures for insectivorous birds and mammals are calculated.

General risk management measures based on EU waste legislation constitute appropriate means for containment of substances ending up in landfill sites; this route of exposure is not considered to be of concern. Recovery of the active substance following disposal is not envisaged or deemed practicable.

Due to the low volume of the a.s. imported to the EU, no estimation of regional or continental PECs is conducted as they are deemed negligible compared to the exposure on local scale caused by professional use.

2.2.2.5. Risk characterisation

For margosa extract the only stage of the life-cycle that is regarded as relevant is ‘emission from professional use’, whilst the other stages, i.e., production (outside the EU), formulation (negligible emission taking into account the described manufacturing processes), private use (not intended), service life (does not apply in regard of the intended use), and waste disposal (covered by measures based on EU waste legislation), are considered not relevant in the present evaluation.

Emission scenarios for outdoor application of insecticides are developed for rural as well as urban areas in the OECD draft ESD. Releases occurring from professional use of the b.p. NeemAzal-T/S are relevant for both areas.

The following emission scenarios are addressed for urban as well as for rural environments:

• Mixing/loading

• Spray application including spray drift, deposition and run-off during application as

well as wash-off by rainfall


32

Aquatic Compartment

In urban areas 100 % of the release of the b.p. end up in the sewers and STP. Secondarily affected compartments in the aquatic environment are surface water and sediment.

In rural areas, spray drift to the aquatic environment is possible. Release to surface water was estimated according to FOCUS surface water.

Estimated PEC/PNEC ratios for surface water (rural as well as urban scenario) and for sediment (urban scenario) are above the trigger value. Therefore a risk from the professional use of NeemAzal-T/S to aquatic ecosystems might be expected. However the estimated risk is based on very conservative assumptions. Concerning risk mitigation measures please refer to section 3.3.

Terrestrial Compartment including Groundwater

The intended use of the b.p. leads to a direct release of the a.s. margosa extract to soil in rural areas and therefore into groundwater as well. For urban scenarios direct release to soil is not supposed to occur. Indirect exposure to soil und subsequently to groundwater is given by application of sewage sludge.

For the soil compartment a PEC/PNEC ratio of 1.04 was determined for the scenario run-off/wash-off. Although the trigger value of 1 is slightly exceeded, the risk is nevertheless regarded as acceptable, as the PNECsoil is a conservative value. The NOEC from the soil micro-organism study, which is the basic value for the PNECsoil, was the highest tested concentration. Therefore, it can be assumed that the “real” NOEC from this test might be higher. In addition, the PECsoil was calculated without considering any degradation thus also representing a highly conservative situation. For all other scenarios, no risk could be identified as the PEC/PNEC ratios are below the trigger value.

For the groundwater assessment, aerobic degradation of azadirachtin A in soil was taken into account. The DT50 values for azadirachtin A were calculated to range from 3.6 to 71.9 days at an average EU outdoor temperature of 12 °C; the geometric mean of 10.34 days was used for PECgroundwater calculation. With this assumption the concentration in groundwater falls below the trigger value of 0.1 µg/L as laid down in Directive 98/83/EC. Therefore, no risk for groundwater is expected from the intended use of NeemAzal T/S.

Only a qualitative assessment for both bees and non-target arthropods is performed as no quantitative risk assessment approach exists in the EC TGD on Risk Assessment (2003).

For honeybees a qualitative risk assessment was performed by comparing the intended application rate of margosa extract (14.13 g a.s./ha) with an application rate from a semi-field study resulting in no effects (176 g a.s./ha). From the factor of 12 between these values it can be assumed that the risk for honeybees resulting from the intended application of margosa is acceptable.


33

In several extended laboratory tests and (semi-)field studies with other non-target arthropods, Chrysoperla carnea, Cocinella septempunctata and Episyrphus balteatus were found to be most sensitive. In extended laboratory studies, the lowest effect value was found for Chrysoperla carnea with a LR50 of about 0.77 g azadirachtin A/ha; at 1.9 g azadirachtin A/ha, mortality was 100 %. Aged residue studies show high mortality with freshly applied test substance (48.4 g azadirchatin A/ha) but the mortality decreased to 10.3 % with residues aged for 14 d. For Episyrphus balteatus NOER values of < 57.6 g Aza A/ha and < 30 g Aza A/ha were obtained in semi-field studies.

For a qualitative risk assessment, the dose on the oak leaves after application (as g a.s/ha leaf area) is compared with the available effect values for non-target arthropods. Doses of 9.49 g azadirchatin A/ha leaf area (mean) or 12.00 g azadirachtin A/ha leaf area (worst-case) were calculated.

A comparison of these exposure rates with the effect values for azadirachtin A results in a clear risk for sensitive non-target arthorpods living on the leaves of the treated trees. For the most sensitive species tested in extended laboratory test (Chrysoperla carnea, Cocinella

septempunctata), LR50 values are in the range of the exposure rates or even below. Therefore, a risk for non-target arthropods has to be assumed from the intended use of margosa extract.

However, a fast recovery of non-target arthropods is expected for the following reasons: Photodegradation, which is known to be fast for azadirachtin A, is not taken into account in the calculation of the exposure rates. In effect studies that used aged residues, it was shown that the toxicity of margosa extract respectively azadirachtin A significantly declines when arthropods are exposed to aged residues compared to freshly applied test substance. Therefore, after a certain period, the content of margosa extract on the oak leaves decreases to a level that does not pose a risk for non-target arthropods. Considering that normally not a total area but only single trees are treated with margosa extract, there seems to be a potential for recolonisation of treated oak trees from surrounding vegetation after short time. In addition, a recovery of non-target arthropod population in treated oak trees seems possible, as for the exposure calculations several conservative assumptions were made.

Secondary Poisoning

Although margosa extract has only a low bioaccumulation potential, insectivorous birds and small mammals may be exposed to the substance when feeding on treated caterpillars in and beneath newly sprayed oak canopies. The assessments of secondary poisoning for small, insectivorous birds and small, insectivorous mammals do not indicate a risk (PEC/PNEC < 1). Therefore the use of margosa extract does not present a risk for secondary poisoning.

2.2.3. List of endpoints

In order to facilitate the work of Member States in granting or reviewing authorisations, and to apply adequately the provisions of Article 5(1) of Directive 98/8/EC and the common principles laid down in Annex VI of that Directive, the most important endpoints, as identified during the evaluation process, are listed in Appendix I.


34

3. DECISION

3.1. Background to the Decision

Article 10 of the Biocides Directive 98/8/EC addresses the inclusion of an active substance in the Annexes I, IA or IB. For the decision of inclusion or non-inclusion, it has to be examined if the criteria of article 10 (1) are fulfilled.

The insecticide NeemAzal-T/S is an emulsifiable concentrate containing further ingredients beside the active substance margosa extract. Due to the nature of the biocidal product it is not expected to exhibit any hazardous physical-chemical properties. It was noted that due to the limited thermal stability of the limonoids in margosa extract, the product may show limited shelf life, which can possibly be solved by recommending storage conditions (maximum temperature specified) and/or end use date.

The analytical methods for NeemAzal-T/S in all relevant matrices are covered by the methods of azadirachtin A. Due to the complex nature of margosa extract, the choice of azadirachtin A as lead compound for residues in soil and water is accepted. The available data on analytical methods for determination of residues are considered sufficient to support an Annex I inclusion of margosa extract.

The studies provided to support the efficacy of margosa extract confirm the efficacy of margosa extract and margosa extract containing formulations against the caterpillars of Thaumetopoea processionea.

The effects on human health have been assessed, in accordance with the provisions of Article 10(1) of Directive 98/8/EC, for the uses proposed by the applicant. The available data on mammalian toxicology and mutagenicity are considered to adequately support the risk evaluation of margosa extract in humans. Concerning toxicology and metabolism the studies required by Directive 98/8/EC are available or statements for non submission have been accepted.

Based on the results observed in the developmental toxicity study, it is proposed to classify margosa extract as sensitising by skin contact (Xi, R 43) and as toxic to reproduction Category 3 (Cat. 3, Xn, R 63).

If the margosa extract containing product NeemAzal-T/S is applied in accordance with the intended use, no residues of margosa extract are to be expected in food of plant or animal origin. Furthermore, the use of the product is foreseen for professional use only. Therefore, primary exposure to non-professionals is not expected. For secondary exposure a human health risk assessment of margosa extract was performed. According to the outcome of the risk assessment, it can be concluded that secondary exposure to margosa extract of the general public (adults and infants including children) is considered acceptable.

The occupational risk assessment for margosa extract is based upon the AEL approach and the estimate of potential occupational exposure. The risk assessment is considered to be sufficiently comprehensive and reliable for the purposes of Annex I inclusion of margosa extract. For the application of the biocidal product the risk assessment does not lead to concern. It is essential to recognize that this conclusion only applies to the active substance


35

(margosa extract) in the biocidal product. From the point of view of occupational safety and health there is no risk-related reason for conditioning the requested Annex I inclusion for margosa extract.

However, against the background of experimental evidence of developmental toxicity, it needs to be stressed, that the AEL for margosa extract should be adhered to on every single day of an exposure period.

On the basis of the risk assessment done for the different environmental compartments, the conclusion was reached that the biocidal product Neem Azal-T/S containing the active substance margosa extract at a concentration of up to 3.19 % w/w may pose a risk to surface water, sediment and non-target arthropods if used as an insecticide to control early larval stages of the oak processionary moth, Thaumetopoea processionea, under outdoor conditions (MG03 / PT18). However the assumptions taken for the environmental exposure assessment are considered very conservative as, for example, no degradation was taken into account although azadirachtin A photodegrades very rapidly in air or the input parameter set in the emission scenarios represent altogether conservative exposure situations.

For the risk assessment of margosa extract, an active substance of natural origin that degrades rapidly and does not have the potential for bioaccumulation, it has to be taken into account that the treatment of infested trees is necessary to prevent allergic reactions of inhabitants to hairs of the oak processionary moth, i.e. protection of human health. It needs to be considered that the use of margosa extract against the oak processionary moth should be restricted to the 1st and 2nd larvae stage which limits the extent of application considerably.

Especially in large wood land areas an infestation is not treated but the access to the area is restricted. Therefore the influence of margosa extract to biodiversity is spatially restricted.

If a treatment of an oak processionary moth infestation with margosa extract is necessary for the protection of human health, adequate risk mitigation measures should be taken into account (see in detail in chapter 3.3). For margosa extract the persistence criteria of soil and surface water according to Annex VI of Directive 98/8/EC, further explained in Technical Notes for Guidance on Annex I Inclusion, chapter 5.3, are not fulfilled based on the degradation results of azadirachtin A and azadirachtin B, which are both major constituents of the a.s. However, due to the chemical nature of margosa extract the trigger values for non-extractable residues could not be checked. Considering all available environmental information margosa extract is in compliance with the environmental criteria for approval of active substance according to Annex VI of Directive 98/8/EC.

3.2. Decision regarding Inclusion in Annex I

The active substance margosa extract shall be included in Annex I to Directive 98/8/EC as an active substance for use in product-type 18 (insecticides, acaricides and products to control other arthropods), subject to the following specific provisions:


36

The active substance margosa extract, is defined as an extract of the kernels of Azadirachta

indica extracted with water and further processed with organic solvents. Minimum purity of margosa extract is 100 %.

When assessing the application for authorisation of a product in accordance with Article 5 and Annex VI, Member States shall assess, where relevant for the particular product, the populations and environmental compartments that may be exposed to the product and the use or exposure scenarios that have not been representatively addressed at the Union level risk assessment.

Due to the identified risks for surface water, sediment, and non-target arthropods, the product shall only be authorised applying several risk mitigation measures (for details see chapter 3.3).

Justification for the Provisions regarding the Inclusion on to Annex I

The composition of the margosa extract depends on the manufacturing method and the used solvents for extraction. However, this information is confidential. Therefore, the description of identity of margosa extract for Annex I inclusion has to be a more general term. Nevertheless, the evaluation of margosa extract is based on this specific composition and does not cover margosa extracts with basically other compositions.

Due to the nature of the extract, the whole extract is the active substance.

3.3. Elements to be taken into account by Member States when authorising products

Some information for the insecticide NeemAzal-T/S was not submitted by the participant within the procedure of inclusion of an active substance in Annex I of Directive 98/8/EG. For the decision of inclusion of margosa extract in Annex I, this information is dispensable, but for product authorization mandatory. In detail the following data/information have to be submitted for product authorisation:

1. Information submitted for the life cycle stages “production of the a.s.” and “formulation of the b.p.” is highly specific for the applicant Trifolio-M GmbH: The a.s. is produced outside the EU. The formulation of the b.p. is carried out in a dedicated processing system resulting in negligible releases to the environment; all processing wastes are disposed of as hazardous waste. Within the scope of the product authorisation it should be checked whether the production and formulation processes as described by the applicant Trifolio-M GmbH still apply, in particular if a different company submits an application for product authorisation.

2. The amount of aflatoxins (sum of B1, B2, G1 and G2) should not exceed 100 µg/kg margosa extract (based on the FAO specification of 300 µg/kg azadirachtin A).

3. The information provided regarding the effectiveness against Thaumetopoea

processionea is only sufficient to show a basic efficacy of margosa extract. Adequate statistically protected test reports regarding the effectiveness have to be provided at the


37

stage of product authorization. Also tests showing the reduction of stringing hairs from caterpillars in the air beneath treated oaks should be provided.

4. Should applications be made for authorisation of products containing margosa extract that may lead to residues in food or feed, Member States shall verify the need to set new or to amend existing maximum residue levels (MRLs) according to Regulation (EC) No 470/2009 or Regulation (EC) No 396/2005, and take any appropriate risk mitigation measures ensuring that the applicable MRLs are not exceeded.

Biocidal products to control the caterpillars of the oak processionary moth (Thaumetopoea

processionea) should only be authorized for professional use because special expertise is needed as the treatment has to be performed exclusively in the 1st or 2nd larval instar.

Due to the identified risks for surface water, sediment, and non-target arthropods, risk mitigation measures should be taken into account. In addition to the usual measures to protect the environment as precaution during handling and prevention of spilling the following specific risk mitigation measured should be complied with when using margosa extract against the oak processionary moth:

• No precautionary treatment of possible infestations with the oak processionary moth with products containing margosa extract,

• Treatment only by trained professionals after a severe infestation was identified that cannot be treated by alternative measures

• Treatment exclusively in the 1st or 2nd larval instar • Treatment only at adequate weather conditions (only light wind, no rain forecasted for

the next days) • Minimal distances from water bodies should be set • No spraying in the direction to surface water. • Considering the potential risk for non-target arthropods when using a product

containing the broad-spectrum insecticide margosa extract, the extent of the campaign needed to control the oak processionary moth should be assessed in relation to the health related nuisance.

3.4. Requirement for further information

It is considered that the evaluation has shown that sufficient data have been provided to verify the outcome and conclusions, and permit the proposal for the inclusion of margosa extract in Annex I to Directive 98/8/EC.

3.5. Updating this Assessment Report

This assessment report may need to be updated periodically in order to take account of scientific developments and results from the examination of any of the information referred to in Articles 7, 10.4 and 14 of Directive 98/8/EC. Such adaptations will be examined and finalised in connection with any amendment of the conditions for the inclusion of margosa extract in Annex I to the Directive.


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Appendix I: List of endpoints

Chapter 1: Identity, Physical and Chemical Properties, Further Information, and

Proposed Classification and Labelling

Active substance (Common Name, not an ISO name)

Margosa extract

Function (e.g. fungicide) Insecticide

Rapporteur Member State Germany

Identity (Annex IIA, point II.)

Chemical name (IUPAC) Not applicable (active substance is an extract)

Chemical name (CA) Margosa, extract.

CAS-No 84696-25-3

EC No 283-644-7

Other substance No Not applicable (active substance is an extract)

Minimum purity of the active substance as manufactured (g/kg or g/l)

Active substance is an extract, purity hence 100%.

Identity of relevant impurities and additives (substances of concern) in the active substance as manufactured

Aflatoxines B1 (main compound), B2, G1, G2

≤ 100 µg/kg

Molecular formula Not applicable (active substance is an extract)

Molecular mass Not applicable (active substance is an extract)

Structural formula Not applicable (active substance is an extract)


Physical and chemical properties (Annex IIA, point III, unless otherwise indicated)

Melting point (state purity) Margosa extract partially liquifies above 120 °C and decomposes above 200 °C (purity 100 % margosa extract)

Boiling point (state purity) The boiling point of margosa extract cannot be observed since decomposition occurs already during melting.

Temperature of decomposition Margosa extract partially liquifies above 120 °C and decomposes above 200 °C

Appearance (state purity) Margosa extract technical is a pale yellow to light brownish powder with garlic like odour (purity 100 % margosa extract )

azadirachtin A is a white odourless powder.

Relative density (state purity) D204 = 1.340 at 20 °C (purity 100 % margosa

extract)

Surface tension Test not applicable because no saturated test solution with the same ratio of components as in margosa extract could be produced.

Vapour pressure (in Pa, state temperature) No test conducted (extraction mixture). Based on the calculated vapour pressure of 3.6·10-13 Pa for azadirachtin A the vapour pressure of the extraction mixture should be << 10-5 Pa

Henry’s law constant (Pa m3 mol -1) Not applicable (extraction mixture)

Solubility in water (g/l or mg/l, state temperature)

Test not conducted (extraction mixture)

solubility of azadirachtin A: 2.9 g/L at 20 °C

Solubility in organic solvents (in g/l or mg/l, state temperature) (Annex IIIA, point III.1)

acetone: >200 g/L dichloromethane: >200 g/L

n-hexane: 1.83 g/L 2-propanol: 51.5 g/L

ethylacetate: >200 g/L toluene: 15.5 g/L

at 20°C; values to be considered as indicative only.

Stability in organic solvents used in biocidal products including relevant breakdown products (IIIA, point III.2)

Stability tests suggest the active substance to be acceptably stable.


40

Partition coefficient (log POW) (state temperature)

Test not applicable (extraction mixture)

Hydrolytic stability (DT50) (state pH and temperature) (point VII.7.6.2.1)

pH 4: 60.5 hours at 40 °C (112.7 d at 12 °C)

pH 7: 30 hours at 40 °C (40.9 d at 12 °C)

pH 8: 5 hours at 40 °C (8.2 d at 12 °C)

Dissociation constant (not stated in Annex IIA or IIIA; additional data requirement from TNsG)

Test not required (extraction mixture)

UV/VIS absorption (max.) (if absorption > 290 nm state ε at wavelength)

Plateau of absorbance between 200 to 220 nm with an extinction coefficient of about 11.5 x 106 cm2 mole –1

Photostability (DT50) (aqueous, sunlight, state pH) (point VII.7.6.2.2)

The half-life for margosa extract was not determined. It was calculated to be 1.8 months and 5.5 days for solar radiation intensities in January or July, respectively, assuming standard degradation behaviour at 55 ° latitude.

Quantum yield of direct phototransformation in water at Σ > 290 nm (point VII.7.6.2.2)

5.55 x 10-4

Flammability Margosa extract is not a highly flammable solid. No self-ignition temperature was observed up to a maximum test temperature of 403 °C.

Explosive properties Margosa extract is not explosive


Classification and proposed labelling (Annex IIA, point IX.)

with regard to physical/chemical data None required

with regard to toxicological data According to the criteria in Dir. 67/548/EEC:

Xi R43 (May cause sensitisation by skin contact)

Xn Repr. Cat. 3 R 63 (Possible risk of harm to the unborn child)

According to the criteria in Reg. 1272/2008:

H317 - May cause an allergic skin reaction

H361d - Suspected of damaging fertility or the unborn child

with regard to fate and behaviour data R 53 (may cause long-term adverse effects in the aquatic environment)

with regard to ecotoxicological data R 51 (toxic to aquatic organisms)


Chapter 2: Methods of Analysis

Analytical methods for the active substance

Technical active substance (principle of method) (Annex IIA, point 4.1)

The methods for determination of azadirachtin A is based on HPLC separation with UV detection.

Impurities in technical active substance (principle of method) (Annex IIA, point 4.1)

The impurities are analysed by HPLC/fluorescence detection of their derivatives. The methods are described in Document III-A 4.1.

Analytical methods for residues

Soil (principle of method and LOQ) (Annex IIA, point 4.2)

Azadirachtin A HPLC-UV 0.02 mg/kg (standard soil) The method is based on liquid extraction, followed by clean up and determination using HPLC with UV detection.

LC-MS/MS 0.02 mg/kg (standard soil) Soil samples were extracted with methanol and cleaned up via solid phase extraction on C18 phase and amino phase. Final determination was performed by LC/MS/MS, monitoring two parent-daughter ion transitions for quantitation and confirmation (703 � 585 amu; 703 � 685 amu).

Air (principle of method and LOQ) (Annex IIA, point 4.2)

Azadirachtin A LC-MS/MS 3 µg/m³ (ambient air, warm humid air)

Air is drawn through XAD adsorption tubes. The adsorption material is extracted with acetonitrile. Final determination was performed by LC/MS/MS, monitoring two parent-daughter ion transitions for quantitation and confirmation (703 � 567 amu; 703 � 585 amu).

Water (principle of method and LOQ) (Annex IIA, point 4.2)

Azadirachtin A HPLC-UV 1 µg/L (surface water) The method is based on solid-phase extraction followed by clean up and determination using HPLC with UV detection.

LC-MS/MS 0.05 µg/L (drinking water, surface water) The method is based on solid-phase extraction. Final determination was performed by LC/MS/MS, monitoring two parent-daughter ion transitions for quantitation and quantitative confirmation (703 � 567 amu; 703 � 585 amu).

Body fluids and tissues (principle of method Not relevant as not classified as toxic or highly


43

and LOQ) (Annex IIA, point 4.2) toxic

Food/feed of plant origin (principle of method and LOQ for methods for monitoring purposes) (Annex IIIA, point IV.1)

Not relevant as no residues expected

Food/feed of animal origin (principle of method and LOQ for methods for monitoring purposes) (Annex IIIA, point IV.1)

Not relevant as no residues expected


Chapter 3: Impact on Human Health

Absorption, distribution, metabolism and excretion in mammals (Annex IIA, point 6.2)

Rate and extent of oral absorption: No data, technically not feasible

100 % oral absorption by default

Rate and extent of dermal absorption: 100 % dermal absorption by default

Distribution: No data, technically not feasible

Potential for accumulation: No data, technically not feasible

Rate and extent of excretion: No data, technically not feasible

Toxicologically significant metabolite No data, technically not feasible

Acute toxicity (Annex IIA, point 6.1)

Rat LD50 oral > 5000 mg/kg bw

Rat LD50 dermal > 2000 mg/kg bw

Rat LC50 inhalation > 0.72 mg/L (4-h exposure, whole body, max. attainable conc.)

Skin irritation Not irritating

Eye irritation Not irritating

Skin sensitization (test method used and result)

Sensitising (M+K)

Repeated dose toxicity (Annex IIA, point 6.3)

Species/ target / critical effect Rat: liver and thyroid (organ wt and histological changes)

Lowest relevant oral NOAEL / LOAEL NOAEL 13-wk, rat: 32 mg/kg bw/d (400 ppm)

Lowest relevant dermal NOAEL / LOAEL No data, not required

Lowest relevant inhalation NOAEL / LOAEL No data, not required

Genotoxicity (Annex IIA, point 6.6) No evidence of a genotoxic potential in humans


45

Chronic toxicity and Carcinogenicity (Annex IIA, point 6.5/6.7)

Target / critical effect No adverse effect reported in rats up to the highest dose tested

Lowest relevant NOAEL / NOEL NOAEL 2-yr, rat: 448 mg/kg bw/d (6400 ppm)

Carcinogenicity No evidence of a carcinogenic potential in rats

Mouse: no data, justification accepted

Reproductive toxicity (Annex IIA, point 6.8)

Species/ Reproduction target / critical effect No effects on reproductive performance up to the highest dose tested

Lowest relevant reproductive NOAEL / LOAEL

NOAEL parental: 50 mg/kg bw/d (750 ppm )

NOAEL reproduction: 50 mg/kg bw/d (750 ppm )

NOAEL offspring: 50 mg/kg bw/d (750 ppm )

Species/Developmental target / critical effect Rat: heart (malformations: interventricular septal defects and malrotation) and ↑ incidence of supernumerary ribs

Rabbit: ↓ number and size of live litters, ↑ resorptions at maternal toxic doses

Lowest relevant developmental NOAEL / LOAEL

NOAEL maternal: 225 mg/kg bw/d (rat)

NOAEL maternal: 20 mg/kg bw/d (rabbit)

NOAEL developmental: 50 mg/kg bw/d (rat)

NOAEL developmental: 100 mg/kg bw/d (rabbit)

Neurotoxicity / Delayed neurotoxicity (Annex IIIA, point VI.1)

Species/ target/critical effect Chicken: no evidence for delayed neurotoxicity (non-guideline study)

Lowest relevant developmental NOAEL / LOAEL.

NOAEL: 1000 mg/kg bw/d

Other toxicological studies (Annex IIIA, VI/XI)

No data available, not required


46

Medical data (Annex IIA, point 6.9)

Open literature and medical surveillance report in production of margosa extract in India

Intoxications and mortalities are reported with neem derived material, specifically with oil and leaves, but not with margosa extract.

No effects in manufacturing staff in three year observation period reported.


Summary (Annex IIA, point 6.10)

Margosa extract Value Study Safety factor

AEL acute*

0.5 mg/kg bw /d

Teratogenicity, rat

100

AEL medium-term*

0.32 mg/kg bw /d

90-d, rat

100

AEL long-term*

0.32 mg/kg bw /d

90-d, rat

100

Drinking water limit Not necessary, not allocated

ADI (if residues in food or feed) Not allocated, no residues in food or feed

ARfD (acute reference dose) Not allocated, no residues in food or feed

Professional user

Reference value for inhalation (proposed OEL)

not determined

Reference value for dermal absorption not determined

* AEL: Systemic (= Internal) Acceptable Exposure Level

Azadirachtin A Value Study Safety factor

AEL acute* 0.2 mg/kg bw /d$ Teratogeni-

city, rat 100

AEL medium-term* 0.09 mg/kg bw /d$ 90-d, rat 100

AEL long-term** 0.09 mg/kg bw /d$ 90-d, rat 100

Drinking water limit Not necessary, not allocated

ADI (if residues in food or feed) Not allocated, no residues in food or feed

ARfD (acute reference dose) Not allocated, no residues in food or feed

Professional user

Reference value for inhalation (proposed OEL)

not determined

Reference value for dermal absorption not determined

* AEL: Systemic (= Internal) Acceptable Exposure Level


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$ The test material used for the teratogenicity study in rats contained 36.6 % azadirachtin A, while for the 90-d study in rats material containing 27.6 % azadirachtin A was used (analytical findings from the respective study reports, otherwise both test materials were generated by the identical technical procedure).


49

Acceptable exposure scenarios (including method of calculation)

Professional users

Production of active substance: n.a.

Formulation of biocidal product Based on exposure evaluation with EASE Model (EASE for Windows Version 2.0, 1997) the exposure estimate is below the AEL. The results of the exposure assessment are not further assessed in the risk characterisation under the requirements of the BPD.

Intended uses Control of 1st and 2nd larval stage (caterpillar) of the oak procession moth, Thaumetopoea processionea, at 1 to maximum 2 applications per year targeted onto infested foliage and branches using a hand-held pump spray device.


50

Mixing & loading:

45 mL NeemAzal T/S (3 % a.s.) is diluted with water in batches of 15 L - final concentration is 0.009 % a.s.

Form of exposure: liquid (3 % a.s.)

Duration: 5-10 min

Frequency: 8 weeks or at most 48 days per year

Model: German Model BBA

Application:

Spraying of 0.009 % a.s., 10 L per tree

(approximately 10 - 20 trees may be treated per day)

Form of exposure: aerosol (0.009 % a.s.)

Duration: 30 – 60 min. per task


Model: German Model BBA

Post-application:

Reloading of the spraying device, cleaning of spraying device

Form of exposure: liquid (0.009% a.s.)

Duration: 30 – 60 min.


Model: Riskofderm (deo unit 1)

Inhalation exposure

Dermal exposure

(all phases)

Not applicable

(no aerosol formation)

10.7

Secondary exposure Since exposure was estimated to be well below the AEL no calculations or measurements are necessary to conclude that exposure of bystanding professionals will be below exposure limits.

Non-professional users Not applicable, however, if comparable products for non-professionals will be placed on the market exposure is considered acceptable (German Model, House & Garden).

Indirect exposure as a result of use General Public, Bystander: Exposure was estimated according to Martin et al. (2008). Exposure of bystanding infants and adults was below AELmedium-

term.

General Public, Re-entry: Exposure was estimated according to Martin et al. (2008). Exposure of re-entering infants and adults was below AELmedium-


51

term.

No residues in food or feed expected.


Chapter 4: Fate and Behaviour in the Environment

Route and rate of degradation in water (Annex IIA, point 7.6, IIIA, point XII.2.1, 2.2)

Hydrolysis of active substance and relevant metabolites (DT50) (state pH and temperature)

Azadirachtin A, measured values at 40 °C

pH 4 : 60.5 hours (117.7 d at 12 °C)

pH 7: 30 hours (40.9 d at 12 °C)

pH 8: 5 hours (8.2 d at 12 °C)

Metabolites are not detectable due to technical limitations

Photolytic / photo-oxidative degradation of active substance and resulting relevant metabolites

Photolytic half-life calculated by means of ABIWAS (latitude 55°N) based on quantum yield (Φc

E = 5.55 x 10-4) and UV/VIS data from direct photo transformation test in water of margosa extract :

January : 1.8 months

July : 5.5 days

Readily biodegradable (yes/no) No

Biodegradation in seawater Not relevant

Non-extractable residues Not feasible

Distribution in water / sediment systems (active substance)

Not feasible

data from literature available for unlabelled azadirachtin A, but only regarded as additional information

Distribution in water / sediment systems (metabolites)

Not feasible

Route and rate of degradation in soil (Annex IIIA, point VII.4, XII.1.1, XII.1.4; Annex VI, para. 85)

Mineralization (aerobic) Not feasible

Laboratory studies (range or median, with number of measurements, with regression coefficient)

DT50lab (20°C, aerobic):

Azadirachtin A:

German standard soils 2.1, 2.2 & 2.3 (sand, loamy sand, sandy loam)


53

Single 1storder: DT50: 1.9 – 4.0 d; n = 4, 20°C )

r2 (range) = 0.955 – 0.997

DT50 geo-mean: 2.6 d (n=4)

Converted to average EU outdoor temp. of 12 °C:

DT50 geo-mean: 4.9 d (n=4, 3.6 –7.6 d)

A further degradation study submitted under PPP legislation was taken into account resulting in a summarised geo mean DT50 of 5.45 days (n=6) at 20°°°°C, corresponding to 10.34 days (n=6) at 12°°°°C.


Azadirachtin A:

Single 1storder: DT90: 6.5 – 13.4 d; (n = 4, 20°C )

r2 (range) = 0.955 – 0.997

geo mean DT90: 8.6 d (n=4)

A further degradation study submitted under PPP legislation was taken into account resulting in a summarised geo mean DT90 of 18.15 days (n=6) at 20°°°°C.


Not applicable

DT50lab (20°C, anaerobic):

This is not expected to be a relevant route of degradation.

Degradation in the saturated zone:

Not applicable

Field studies (state location, range or median with number of measurements)

DT50f:

Not relevant in regards of the laboratory findings.

DT90f:

Not relevant in regards of the laboratory findings.

Anaerobic degradation This is not expected to be a relevant route of degradation.

Soil photolysis Not feasible;


54

In literature a DT50 value of 3.87 d on glass is published.

Non-extractable residues Not applicable

Relevant metabolites - name and/or code, % of applied a.i. (range and maximum)

Not applicable

Soil accumulation and plateau concentration Not required

Adsorption/desorption (Annex IIA, point XII.7.7; Annex IIIA, point XII.1.2)

Ka , Kd

Kaoc , Kdoc

pH dependence (yes / no) (if yes type of

dependence)

Azadirachtin A:

Ka : 0.373, 0.405, 0.479 (n = 3)

Kaoc: 20.6 mL/g, 30.6 mL/g, 65.4 mL/g (n = 3)

No

A further study submitted under PPP legislation was taken into account resulting in a median value of 75.2 mL/g (ntotal = 7).

Fate and behaviour in air (Annex IIIA, point VII.3, VII.5)

Direct photolysis in air Not relevant

Quantum yield of direct photolysis Not relevant

Photo-oxidative degradation in air Atkinson method, AOP model (24-h average concentration of 5.0 x 105 OH radicals cm-3)

Azadirachtin A, half-life in air : 0.071 d or 1.696 h

Volatilization Not expected in regards of vapour pressure of 3.6 x 10-13 Pa for azadirachtins

Monitoring data, if available (Annex VI, para. 44)

Soil (indicate location and type of study) No data available

Surface water (indicate location and type of study)

No data available


55

Ground water (indicate location and type of study)

No data available

Air (indicate location and type of study) No data available


56

Chapter 5: Effects on Non-target Species

Toxicity data for aquatic species (most sensitive species of each group) (Annex IIA, point 8.2, Annex IIIA, point 10.2)

Species Time-scale Endpoint Toxicity (mg a.s./L) 1)

Fish

Oncorhynchus mykiss 96-h, semi-static,

Mortality, LC50 4.1 2)

Oncorhynchus mykiss 28-d, flow-through

Mortality, growth,

NOEC / LOEC

1.9 / 4.4 2)

Invertebrates

Daphnia magna 48-h, static Immobility, EC50 9.69

Daphnia magna 21-d, semi-static

Reproduction, NOEC / LOEC

0.1 / 0.222)

Chironomus riparius 28 d static Emergence, development rate NOEC

0.0062)

Algae

Scenedesmus subspicatus 72-h, static Growth rate, ErC50

Biomass, EbC50

Growth rate, ErC10

1041

482

332

Microorganisms

Activated sludge predominately from municipal wastewater treatment plant

30 min, static

Respiration inhibition (oxygen consumption),

NOEC

1000

1) a.s. refers to margosa extract; 2) test conducted with NeemAzal-T/S containing 4% margosa extract

Effects on earthworms or other soil non-target organisms

Acute toxicity to earthworms (Annex IIIA, point XIII.3.2 ; Annex IIIB, point XIII.3.4)

LC50 > 12 mg a.s./kg dry soil

(test conducted with product NeemAzal-T/S containing approx. 1.2% margosa extract)

Reproductive toxicity to earthworms or other soil non-target macroorganisms

Hypoaspis aculeifer : 14d-NOEC = 5974 mg a.a./kg


57

(Annex IIIA, point XIII.3.2) dry soil

Terrestrial plants (Annex IIIA, point XIII.3.4; Annex IIIB, point XIII.3.6)

EC50 > 3.0 L NeemAzal-T/S / ha equivalent to

> 94.5 g a.s./ha corresponds to a soil concentration of > 0.06 mg a.s./kg dw using a soil depth of 10 cm and a soil density of 1500 kg/m³ dry soil

Effects on soil micro-organisms (Annex IIA, point 7.4; Annex IIIB, point XIII.3.5)

Nitrogen mineralization NOEC 39.2 mg NeemAzal-T/S / kg dry soil eqv. to

1.28 mg a.s./kg dry soil

converted to standard soil (2% organic carbon)

NOEC = 2.332 mg a.s./ kg dry soil, equivalent to 2 mg a.s./ kg wet soil

Carbon mineralization NOEC 39.2 mg NeemAzal-T/S / kg dry soil eqv. to

1.28 mg a.s./kg dry soil

converted to standard soil (2% organic carbon)

NOEC = 2.334 mg a.s./ kg dry soil, equivalent to 2 mg a.s./ kg wet soil

Effects on terrestrial vertebrates

Acute toxicity to mammals (Annex IIIA, point XIII.3.3)

refer to chapter 3

Acute toxicity to birds (Annex IIIA, point XIII.1.1)

LD50 > 4000 mg a.s./kg bw

Dietary toxicity to birds (Annex IIIA, point XIII.1.2)

LC50 > 5200 mg a.s./kg feed

Reproductive toxicity to birds (Annex IIIA, point XIII.1.3)

NOEC = 350 mg a.s./kg food*

*The applicant has no access to this study. Therefore, for product authorisation, a letter of access to this study needs to be provided.

Effects on honeybees (Annex IIIA, point XIII.3.1; Annex IIIB, point XIII.3.2)

Acute oral toxicity LD50 > 590 µg NeemAzal-T/S / bee equivalent to


58

> 17.7 µg a.s./bee

Acute contact toxicity LD50 > 2100 µg NeemAzal-T/S / bee equivalent to

> 63 µg a.s./bee


59

Effects on other beneficial arthropods (Annex IIIA, point XIII.3.1; Annex IIIB, point XIII.3.3)

Exposure Species

Design Duration

Endpoint Toxicity (g Aza A/ha) 1)

Laboratory tests

Aphidius

rhopalosiphi

Glass plate test

48 h (M), 11 d (R)

Mortality, LR50

Reproduction, NOER

< 57.6

< 57.6

Typhlodromus pyri Plastic plate test

6 d (M), 12 d (R)

Mortality, LR50

Reproduction, NOER

<30

30

Coccinella

septempunctata

Glass plate test

28 d Mortality, LR50

Reproduction, NOER

> 12

12

Poecilus cupreus Glass plate test

14 d Mortality, LR50

Sublethal effects, NOER

> 8

8

Orius majusculus Plastic plate test

5 d (M), 10 d (R)

Mortality, LR50

Reproduction, NOER

> 50

50

Extended-laboratory tests

Aphidius

rhopalosiphi

Oat seedlings 48 h (M), 11 d (R)

Mortality, LR50

Reproduction, NOER

> 57.6

57.6

Chrysoperla

carnea

Broad bean seedlings

47 d (M, R) Mortality, LR50

Development, NOER

> 60

30

Coccinella

septempunctata

Sweet pepper leaves

19 d (M) 14 d (R)

Mortality, LR50 15.8

Chrysoperla

carnea

Apple leaves 34 d Mortality, LR50, NOER

0.77 < 0.77

Chrysoperla

carnea

Sweet pepper leaves, fresh

and aged residues

38 d Mortality LR50 Reproduction NOER

< 48.4 (freshly applied)

in the treatments

with 14 and 28 days aged

residues mortality was 10.3 % and

3.3 % and no effects on

repro


60

Chrysoperla

carnea

Sweet pepper leaves, fresh

and 21 d aged residues

49 d Mortality LR50

Reproduction NOER

< 20.1 (freshly applied)

in the treatment

with 21 days aged residues mortality was 3.7 % and no

effects on repro

Chrysoperla

carnea

Apple leaves 34 d Mortality LR50

Reproduction NOER

> 1-9

1.9

Cocinella

septempunctata

Bean leaves 47 d Mortality LR50

Reproduction:

1.94

reproduction reduction Dose/reduction: 0.08/ 34.58 % 0.23/ 18.12 % 0.73/ -79.53 % 7/ -108.71 %

Semi-field / Field tests

Typhlodromus pyri Apple trees (field)

117 d (layout A), 94 d (layout B)

Abundance, NOER 42 2)

252 3)

Typhlodromus pyri Grapevines (field)

43 d Abundance, NOER 15.9 4)

31.8 5)

Chrysoperla

carnea

Apple trees (semi-field)

14 d (pup. and em.),

28 d (R)

Mortality, LR50

Reproduction, NOEC

> 60

< 30

Chrysoperla

carnea

Broad bean plants

(semi-field)

34 d (pup. and em.),

37 d (R)

Mortality, LR50

Reproduction, NOER

> 30

30

Coccinella

septempunctata

Sweet pepper plants (field)

42 d (exposure), 89 d (M, R)

Mortality, LR50

Reproduction, NOER

> 50

50

Episyrphus

balteatus

Broad bean plants

(semi-field)

7 d (M), 34 d (R)

Mortality, LR50

Reproduction, NOER

57.6

< 57.6

Episyrphus

balteatus

Broad bean plants

(semi-field)

12 d (M, pup. and hatch.)

Mortality, LR50

Development, NOER

< 30

< 30


61

1) Aza A = Azadirachtin A 2) based on mean application rate of spray sequence of 6 consecutive applications (layout A) 3) based on cumulated application rate of spray sequence of 6 consecutive applications (layout A)

4) based on mean application rate of 2 applications 5) based on cumulated application rate of 2 applications

M: Mortality; R: Reproduction parameters; em.: emergence rate; hatch.: hatching rate; pup.: pupation rate; NOER: No Observable Effect Rate; LOER: Lowest Observable Effect Rate; LR50: median lethal rate


62

Bioconcentration (Annex IIA, point 7.5)

Bioconcentration factor (BCF) Azadirachtin A:

BCFfish = 1.38 (calc.)

BCFearthworm = 1.0 (calc.)

Azadirachtin B:

BCFfish = 2.5 (calc.)

BCFearthworm = 1.1 (calc.)

Depration time (DT50)

(DT90)

Not applicable

Level of metabolites (%) in organisms accounting for > 10 % of residues

Not applicable

Chapter 6: Other End Points

No further endpoints are available.


63

Appendix II: List of Intended Uses

The intended use of the representative insecticide (PT 18) NeemAzal T/S is for professional application, only e.g. in landscape conservation units. NeemAzal T/S is an emulsifiable concentrate which is diluted with tap water by the end user. The intended concentration in the final spray solution is 0.3 % NeemAzal T/S. For the control of the oak procession moth, NeemAzal T/S is applied with a hand-held spraying device from a lifting platform on crown level of the treated trees. The application is targeted onto the foliage and focuses on branches with nests of caterpillars of target organisms in the upper part of the tree.

The submitted studies indicate efficacy of margosa extract against caterpillars of the oak procession moth, Thaumetopoea

processionea. The efficacious dilution used in the field test was 1 % NeemAzal T/S. The efficacy data do not show evidence that in the nature the concentration of 0.3% (the use concentration proposed by the participant) is also active. It has only been shown in the laboratory study that margosa extract is efficacious at a concentration of 0.3%.

Summary of intended uses

combat of insect caterpillars causing allergic reactions in humans and animals

North and south Europe

NeemAzal T/S

Oak procession moth, Thaumetop

oea

processione

a

first and second caterpillar stage

emulsifiable concentrate (EC)

31.4 g/L

spray application

1 to 2 14 days 0.0942 see remarks

see remarks

NeemAzal T/S is applied at a concentration of 0.3 % in a volume of approximately 10 L per tree (approx. 2 – 25 m height). A wetting of the foliage surface is intended. The dilution used in the field test was 1 % NeemAzal T/S.

Object

and/or

situation

Member

State or

Country

Produc

t

name

Organisms

controlled

Formulatio

n Application

Applied amount per

treatment Remarks:

Type

Conc

. of

a.s.

metho

d kind

number

min

max

interval

between

application

s (min)

g

a.s./L

min

max

water

L/m2

min

max

g

a.s./m2

min

max


64

Appendix III: List of studies

Data protection is claimed by the applicant in accordance with Article 12.1(c) (i) and (ii) of Council Directive 98/8/EC for all study reports marked “Y” in the “Data Protection Claimed” column of the table below. For studies marked Yes(i) data protection is claimed under Article 12.1(c) (i), for studies marked Yes(ii) data protection is claimed under Article 12.1(c) (ii). These claims are based on information from the applicant. It is assumed that the relevant studies are not already protected in any other Member State of the European Union under existing national rules relating to biocidal products. It was however not possible to confirm the accuracy of this information.

Reference list of studies on the active substance

Section No /

Reference No

Author(s) Year Title

Source (where different from company)

Company

Report No.

GLP (where relevant)

(Un)Published

Data

Protectio

n

Claimed

(Yes/No)

Owner

A 3.1.1/01* Werle, H. 1995 Melting Point, EC Directive 92/69/EEC,A.1., NeemAzal BioChem GmbH, Karlsruhe, Germany Trifolio-M GmbH Report-no. 955040827 A GLP: yes, GEP: yes Published: no

yes TRF

A 3.1.3/01 Troß, R. 1995a Determination of Pour and Tap Density of NeemAzal [German Original] Trifolio-M GmbH, D-35633 Lahnau Trifolio-M GmbH Report-no. LP 95.2 GLP/GEP: no Published: no

yes TRF

A 3.1.3/02 Kleeberg, H. 2007a Statement conserning the relative density of Margosa extract (CAS-No.: 84696-25-3), variant NeemAzal technical Trifolio-M GmbH, D-35633 Lahnau Trifolio-M GmbH Report-no. not applicable GLP/GEP: no Published: no

yes TRF

A 3.1.3/03 Kabaleeswaran, V., Rajan, S.S., Govindachari, T.R., Gopalakrishnan, G.

1994 Crystal and molecular structure of Azadirachtin-A - Current Science, 66, No. 5, 362-364 Report-no. not applicable GLP/GEP: no Published: yes

no -


65

A 3.1.3/04* Thom, M. 2007 Relative Density of NeemAzal eurofins-GAB GmbH, Niefern-Öschelbronn, Germany Trifolio-M GmbH Report-no. 20071477/01-PCRD GLP: yes Published: no

yes TRF

A 3.2/01* Kleeberg, H. 2005 Vapour pressure of NeemAzal technical Trifolio-M GmbH, D-35633 Lahnau Trifolio-M GmbH Report-no. not applicable GLP/GEP: no Published: no

yes TRF

A 3.2/02 Anonymous 1990 Estimation of vapor pressure Trifolio-M GmbH, D-35633 Lahnau Trifolio-M GmbH Report-no. not applicable GLP/GEP: no Published: no

yes TRF

A 3.2.1/01 Kleeberg, H. 2005 Vapour pressure of NeemAzal technical Trifolio-M GmbH, D-35633 Lahnau Trifolio-M GmbH Report-no. not applicable GLP/GEP: no Published: no See also Doc IIIA 3.2/01

yes TRF

A 3.3/01* Kleeberg, H. 1994a Physico-chemical properties of Azadirachtin A Appearance, Colour and Odour Trifolio-M GmbH, D-35633 Lahnau Trifolio-M GmbH Report-no. LP94.03 GLP/GEP: no Published: no

yes TRF

A 3.3/02 Kleeberg, H. 1994b Physico-chemical properties of NeemAzal technical Appearance, Colour and Odour Trifolio-M GmbH, D-35633 Lahnau Trifolio-M GmbH Report-no. LP94.02 GLP/GEP: no Published: no

yes TRF

A 3.3/03 Kleeberg, H. 2007b Statement to: Azadirachtin: Spectra, a table of signal characteristics and molecular extinction at relevant wavelength for purified active substance Trifolio-M GmbH, D-35633 Lahnau Trifolio-M GmbH Report-no. Enclosure: IIA 2.51 GLP/GEP: no Published: no

yes TRF

A 3.4/01* Kleeberg, H 2006 Azadirachtin: Spectra, a table of signal characteristics and molecular extinction at relevant wavelength for purified active substance Trifolio-M GmbH, D-35633 Lahnau Trifolio-M GmbH Report-no. Enclosure: IIA 2.51 GLP/GEP: no Published: no

yes TRF


66

A 3.5/01 Troß, R. 1995b "SOLUBILITY PROPERTIES OF NEEMAZAL IN WATER[ENGLISH TRANSLATION/ GERMAN ORIGINAL]" Trifolio-M GmbH, D-35633 Lahnau Trifolio-M GmbH Report-no. LP 95.9 GLP/GEP: no Published: no

yes TRF

A 3.5/02 Ruch, B. 2007 Statement to:"SOLUBILITY PROPERTIES OF NEEMAZAL IN WATER[ENGLISH TRANSLATION/ GERMAN ORIGINAL]" Trifolio-M GmbH, D-35633 Lahnau Trifolio-M GmbH Report-no. LP 95.9-Attachment GLP/GEP: no Published: no

yes TRF

A 3.7/01* Troß, R. 1995c Determination of Solubility of NeemAzal in Organic Solvents [German Original] Trifolio-M GmbH, D-35633 Lahnau Trifolio-M GmbH Report-no. LP 95.7 GLP/GEP: no Published: no

yes TRF

A 3.9/01 Troß, R. 1996 PARTITION COEFFICIENTS OF VARIOUS CONSTITUENTS OF NEEMAZAL IN N-OCTANOL/WATER [GERMAN ORIGINAL] Trifolio-M GmbH, D-35633 Lahnau Trifolio-M GmbH Report-no. TM 1195.13 GLP: yes, GEP: yes Published: no

yes TRF

A 3.11/01 Franke, J. 2005a NeemAzal - Flammability (solids) A.10 Siemens AG, Prozess-Sicherheit, Frankfurt am Main, Germany Trifolio-M GmbH Report-no. 20050679.02 GLP: yes Published: no

yes TRF

A 3.11/02 Franke, J.* 2005b NeemAzal - Auto-Flammability (Solids-Determination of Relative Self-Ignition Temperature)A.16 Siemens AG, Prozess-Sicherheit, Frankfurt am Main, Germany Trifolio-M GmbH Report-no. 20050679.03 GLP: yes Published: no

yes TRF

A 3.13/01 Franke, J. 2005c NeemAzal - Surface Tension A.5 (OECD 115) Siemens AG, Prozess-Sicherheit, Frankfurt am Main, Germany Trifolio-M GmbH Report-no. 20050679.01 GLP: yes Published: no

yes TRF


67

A 3.15/01* Smeykal, H. 2002 NeemAzal - Explosive Properties Sicherheitstechnik Siemens Axiva GmbH & Co. KG, Frankfurt Trifolio-M GmbH Report-no. 20020457.01 GLP: yes Published: no

yes TRF

A 3.16/01* Franke, J. 2005d NeemAzal - Oxidizing Properties A.17 Siemens AG, Prozess-Sicherheit, Frankfurt am Main, Germany Trifolio-M GmbH Report-no. 20050679.04 GLP: yes Published: no

yes TRF

A 4.2/01* Troß, R. 1998 METHOD VALIDATION METHOD: RAN-04.01 "DETERMINATION OF AZADIRACHTIN A RESIDUES IN THE SOIL"[GERMAN ORIGINAL] Trifolio-M GmbH, D-35633 Lahnau Trifolio-M GmbH Report-no. LP 98.01 GLP/GEP: no Published: no

yes TRF

A 4.2/02* Troß, R. 1996 VALIDATION OF THE METHOD RAN-03 "DETERMINATION OF AZADIRACHTIN A RESIDUES IN WATER"[GERMAN ORIGINAL] Trifolio-M GmbH, D-35633 Lahnau Trifolio-M GmbH Report-no. LP 96.6 GLP/GEP: no Published: no

yes TRF

A 4.2/03* Ruch, B. 2003 Method Validation Method: Ran-03.OW - Determination of Azadirachtin A-Residues in Surface Water Trifolio-M GmbH, D-35633 Lahnau Trifolio-M GmbH Report-no. V-Ran-03.OW GLP: yes, GEP: yes Published: no

yes TRF

A 4.2/04 Lucini, L. 2007 Specificity of the analytical method RAn-04.1 "Determination of Azadirachtin A-residues in the soil" from Trifolio-M Research Centre "E. Gagliardini" - SIPCAM S.p.A., Salerano sul Lambro (LO), Italy Sipcam S.p.A. / Mitsui AgriScience International / Trifolio-M GmbH Report-no. SIP1569 GLP: yes Published: no

yes SMT


68

A 4.2/05* Class, T. 2007 Development and Validation of an Analytical Method for the Determination of Azadirachtin A in Water, Using LC/MS/MS PTRL Europe, Ulm, Germany Sipcam S.p.A. / Mitsui AgriScience International / Trifolio-M GmbH Report-no. P/B 1305 G GLP: yes Published: no

yes SMT

A4.2/06* Witte, A. 2008 Validation of an analytical method for the determination of Azadirachtin A in soil; testing facility: eurofins-GAB GmbH, Niefern-Öschelbronn, Germany, published: no, report No. 20081004/01-RVS

yes TRF

A4.2/07* Class, T. 2007 Development and validation of an analytical method for the determination of Azadirachtin A in air, PTRL Europe, D-89081 Ulm, Germany report no. P/B 1306 G published: no 20 September 2007

yes TRF SCM

A 5.3.1/01 Breuer. M., de Loof, A.

2000 Laboratory trials with NeemAzal T/S on the allergenic forest pest Thaumetopoea processionea (L.), in: Practice orientated results on use and production of Neem-Ingredients and Pheromones VIII Zoological Institute, Kathol. Universiteit Leuven, Belgium Proc of the 8th workshop, Hohensolms, Germany, 2000 Report-no. GLP/GEP: no Published: yes

no -

A 5.3.1/02 Lehman, M., Fieguth, A.

2000 Further results in using NeemAzal T/S against oak procession moth (Thaumetopoea processionea Linne), in: Practice orientated results on use and production of Neem-Ingredients and Pheromones VIII Pflanzenschutzdienst LA, F. a.d.O. Germany Proc of the 8th workshop, Hohensolms, Germany Report-no. GLP/GEP: no Published: yes

no -

A 5.3.1/03 Hummel, E., Kleeberg, H.

1997 EFFECT OF THE NEEM EXTRACT FORMULATION NEEMAZAL-T/S ON THE GREEN PEA APHID ACYRTHOSIPHON PISUM IN THE LABORATORY (1995)[GERMAN ORIGINAL] not applicable Report-no. not applicable GLP/GEP: no Published: yes

no -


69

A 5.4/01 Kleeberg, H. 2005a Mode of action Trifolio-M GmbH, D-35633 Lahnau Trifolio-M GmbH Report-no. not applicable GLP/GEP: no Published: no

no TRF

A 5.4/02 Rembold, H., Banerjee, S.

1992 Manipulation of insect serotonin pools by azadirachtins not applicable Advances in Tryptophan Research 1992, 125-128 Report-no. not applicable GLP/GEP: no Published: yes

no -

A 5.4/03 Rembold, H. 1995 Growth and metamorphosis not applicable In: The neem tree. H. Schmutterer (ed.), 177-194 Report-no. not applicable GLP/GEP: no Published: yes

no -

A 5.4/04 Banerjee, S., Rembold, H.

1992 Azadirachtin A interferes with control of serotonin pools in the neuroendocrine system of locusts not applicable Naturwissenschaften 79, 81-84 Report-no. not applicable GLP/GEP: no Published: yes

no -

A 5.4/05 Annadurai, R.S., Rembold, H.

1993 Azadirachtin A modulates the tissue specific 2D polypeptide patterns of the desert locust, Schistocerca gregaria not applicable Naturwissenschaften 80, 127-130 Report-no. not applicable GLP/GEP: no Published: yes

no -

A 5.4/06 Rembold, H., Annadurai, R.S.

1993 Azadirachtin inhibits proliferation of Sf 9 cells in monolayer culture not applicable Z. Naturforsch. 48c, 495-499 Report-no. not applicable GLP/GEP: no Published: yes

no -

A 5.4/07 Nisbet, A.J., Mordue, A.J., Mordue, W.

1995 Detection of [22,23-3H2]dihydroazadirachtin binding sites on Schistocerca gregaria (Forskal) testes membranes not applicable Insect Biochem. Molec. Biol. Vol. 25, No. 5, pp. 551-557, 1995 Report-no. not applicable GLP/GEP: no Published: yes

no -


70

A 5.4/08 Iqbal, S., Strang, R.H.C.

1997 The effect of Azadirachtin A on the induction of a specific protein in the locust Sch. Gregaria not applicable Proc of the workshop VII, Wetzlar, 1997 Report-no. not applicable GLP/GEP: no Published: yes

no -

A 5.4/09 Mordue, A.J. 1997 Azadirachtin - a review of its mode of action in insects not applicable Proc of the workshop VII, Wetzlar, 1997 Report-no. not applicable GLP/GEP: no Published: yes

no -

A 5.4/10 Jabbar, A., Strang, R.H.C.

1997 A comparison of the effects of azadirachtin A on cultured insect and mammalian cells not applicable University of Glasgow, G.B. Report-no. not applicable GLP/GEP: no Published: yes

no -

A 5.7/01 Kleeberg, H. 2005b Information on the possible occurence of the development of resistance or cross-resistence not applicable Trifolio-M GmbH Report-no. not applicable GLP/GEP: no Published: no

no TRF

A 5.7/02 Völlinger, M. 1995 Studies of the probability of development of resistance of Plutella Xylostella to neem products not applicable In: The neem tree azadirachta indica A. Juss. And other meliaceous plants. H. Schmutterer (ed.), 477-483 Report-no. not applicable GLP/GEP: no Published: yes

no -

A 5.7/03 Völlinger, M. 1985 STUDIES ON THE RESISTANCE DEVELOPMENT AGAINST INSECTICIDAL COMPOUNDS OF THE NEEM TREE AZADIRACHTA INDICA A. JUSS. IN COMPARISON TO SYNTHETIC INSECTICIDES IN POPULATIONS OF THE CABBAGE MOTH PLUTELLA XYLOSTELLA L. (LEPIDOPTERA, PLUTELLIDAE)[German Original] not applicable Verlag Shaker Report-no. not applicable GLP/GEP: no Published: yes

no -


71

A 5.7/04 Feng, R., Isman, M.B.

1995 Selection for resistance to azadirachtin in the green peach aphid, Myzus persicae not applicable Experientia 51, 831-833 Report-no. not applicable GLP/GEP: no Published: yes

no -

A 5.7/05 Sarupa, M., Krishnaiah, N.V., Reddy, D.D.R.

1999 Insecticide resistance pattern in the selected strains of brown planthopper Nilaparvata Lugens stal in rice not applicable Pesticide Research Journal 11(2), 138-142 Report-no. not applicable GLP/GEP: no Published: yes

no -

A 5.7/06 Abdullah, M., Sarnthoy, O., Tantakom, S., Isichaikul, S., Chaeychomsri, S.

2000 Monitoring insecticide resistance development in beet armyworm, Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae) not applicable Report-no. not applicable GLP/GEP: no Published: yes

no -

(Doc II A2) Gottschling, S & Meyer, S

2006 An Epidemic Airborne Disease Caused by the Oak Processionary Caterpillar

No Published

A 6.1.1/01* 1997a Neemazal technical Acute oral toxicity to the rat Trifolio-M GmbH Report-no. 1749, EIP 6/950799/AC GLP: yes Published: no

yes TRF

A 6.1.2/01* 1997b Neemazal technical Acute dermal toxicity to the rat Trifolio-M GmbH Report-no. 1744, EIP 7/950800/AC GLP: yes Published: no

yes TRF

A 6.1.3/01* 1997 Neemazal technical acute inhalation toxicity in rats 4-hour exposure Trifolio-M GmbH Report-no. EIP 5/951566 GLP: yes Published: no

yes TRF

A 6.1.4/01* 1996a Neemazal technical skin irritation to the rabbit Trifolio-M GmbH Report-no. EIP 8/950822/SE GLP: yes Published: no

yes TRF

A 6.1.4/02* 1996b Neemazal technical eye irritation to the rabbit Trifolio-M GmbH Report-no. 1726, EIP 9/950823/SE GLP: yes Published: no

no TRF


72

A 6.1.5/01* 1997 Neemazal technical skin sensitisation in the guinea-pig Trifolio-M GmbH Report-no. EIP 10/950818/SS GLP: yes Published: no

yes TRF

A 6.2/01* 2005 STUDIES ON THE TRANSDERMAL PENETRATION AND RESORPTION OF NEEMAZAL SHAMPOO AND NEEMAZAL T/S [German Original] not applicable Trifolio-M GmbH Report-no. not applicable GLP/GEP: no Published: no

yes TRF

A 6.3.1/01* 1997 Neemazal technical Toxicity study in rats by dietary administration for 4 weeks Trifolio-M GmbH Report-no. 1720, EIP 3/960397 GLP: yes Published: no

yes TRF

A 6.4.1/01* 1997 Neemazal technical toxicity study in rats by dietary administration for 13 weeks Trifolio-M GmbH Report-no. EIP 4/963100 GLP: yes Published: no

yes TRF

A 6.6.1/01* Jones, E., Gant, R.A.

1997 Neemazal technical bacterial mutation assay Huntingdon Life Sciences Ltd., Huntingdon, UK Trifolio-M GmbH Report-no. EIP 11/950642 GLP: yes Published: no

yes TRF

A 6.6.2/01* Stien, J. 2006 In vitro assessment of the clastogenic activity of NeemAzal in cultured human peripheral lymphocytes LPT Lab. of Pharmacology and Toxicology GmbH & Co KG, Hamburg, Germany Report-no. 19026/1/05 GLP: yes Published: no

yes

A 6.6.3/01* Adams, K., Kirkpatrick, D.

1997 Neemazal technical mammalian cell mutation assay Huntingdon Life Sciences Ltd., Huntingdon, UK Trifolio-M GmbH Report-no. BDP/18, EIP 12/950657 GLP: yes Published: no

yes TRF

A 6.6.4/01* 1997 Neemazal technical Mouse micronucleus test Trifolio-M GmbH Report-no. EIP 13/952782 GLP: yes Published: no

yes TRF


73

A 6.7/01* 2000 Long-term carcinogenicity study of NeemAzal technical in Wistar Rats Trifolio-M GmbH Report-no. 7291 GLP: yes Published: no

yes TRF

A 6.7/02* 1996 Carcinogenicity study of Neemazal-F 5% in mice Trifolio-M GmbH Report-no. 1544/JRF/TOX/96 GLP/GEP: no Published: no

yes TRF

A 6.7/03 Gaitonde, B.B. 1972 EXTRACTS FROM REPORT OF THE SUB-COMMITTEE ON PESTICIDE TOXICOLOGY - Trifolio-M GmbH Report-no. GLP/GEP: no Published: no

no TRF

A 6.8.1/01* 1997a A study of developmental toxicity in rats (gavage administration) Trifolio-M GmbH Report-no. 05-293-97, EIP 2/952493 GLP: yes Published: no

yes TRF

A 6.8.1/02* 1994 A developmental toxicity study of orally administered ATI-720 in rabbits Project No.: L 08424 Study No2b Dates of work: 05.03. – 31.03.1994

yes Certis Europe N.V.

A 6.8.1/03 1997b Neemazal technical - A preliminary study of developmental toxicity in rats (gavage administration) Trifolio-M GmbH Report-no. EIP 2/952493 GLP: yes Published: no

yes TRF

A 6.8.2/01* 2000 Evaluation of Toxicity of NeemAzal Technical to Reproductive Process in Wistar Rats - Segment IV - Toxicity to two Generation Reproductive Process Trifolio-M GmbH Report-no. 4826 GLP: yes Published: no

yes TRF

A 6.8.2/02 Gaitonde, B.B.

1972 EXTRACTS FROM REPORT OF THE SUB-COMMITTEE ON PESTICIDE TOXICOLOGY - Trifolio-M GmbH Report-no. GLP/GEP: no Published: no See also Doc IIIA 6.7/03

no TRF


74

A6.8.2/03 1996 REPRODUCTION TOXICITY STUDY (SEGMENT-IV) OF NEEMAZAL-F 5% IN CHARLES FOSTER RAT unpublished Report-no. 1542/JRF/TOX/96 Trifolio-M GmbH Report-no. 1542/JRF/TOX/96 GLP: yes Published: no

yes TRR

A 6.9/01 1998 Neurotoxicity study with Neemazal technical (27.3% Azadirachtin) in chicken Trifolio-M GmbH Report-no. 4813 GLP/GEP: no Published: no

yes TRF

A 6.12.1/01 Venkataram, T.V.

2002 Employees Health Record 2001 not applicable Trifolio-M GmbH Report-no. not applicable GLP/GEP: no Published: no

yes TRF

A 6.12.1/02 Ventakaram, T.V.

2003 Employess Health Record 2002 not applicable Trifolio-M GmbH Report-no. not applicable GLP/GEP: no Published: no

yes TRF

A 6.12.1/03 Ventakaram, T.V.

2004 Employees Health Record 2003 not applicable Trifolio-M GmbH Report-no. 4826, not applicable GLP/GEP: no Published: no

yes TRF

A 6.12.2/01 Niemann, L., Stinchcombe, S., Hilbig, B.

2002 Toxicity of neem to vertebrates and side effects on beneficial and other ecologically important non-targeted organisms. Toxicity to mammals including humans - The Neem Tree (Azadirachta indica A Juss.) and Other Meliaceous Plants. (Ed. Schmutterer), Weinheim, Germany: VCH, 607-623 Report-no. GLP/GEP: no Published: yes

no -

A 6.12.2/02 Boeke, S.J., Boersma, M.G., Alink, G.M., van Loon, J.J., van Huis, A., Dicke, M., Rietjens

2004 Safety evaluation of neem (Azadirachta indica) derived pesticides - J Ethnopharmacology, 94, 25-41 Report-no. GLP/GEP: no Published: yes

no -

(Doc II A3) Sinniah, D. & Baskaran, G.

1981 MARGOSA OIL POISONING AS A CAUSE OF REYE’S SYNDROME. The Lancet, 1/8218, pp. 487-489

No Published


75

(Doc II A3) Sinniah, D. et al. 1981 MARGOSA OIL POISONING IN INDIA AND MALAYSIA Transactions of the Royal Society of Tropical Medicine and Hygiene. 75/6; pp 903-904

No Published

(Doc II A3) Sundarvalli, N. et al.

1982 NEEM OIL POISONING. The Indian J Pediat. 49/3; pp. 357-359

No Published

(Doc II A3) Sivashanmugham, R. et al.

1984 VENTRICULAR FIBRILLATION AND CARDIAC ARREST DUE TO NEEM LEAF POISONING J. Ass. Phys. India, 32; pp. 610-611

No Published

(Doc II A3) Lai S.M. et al. 1990 MARGOSA OIL POISONING AS A CAUSE OF TOXIC ENCEPHALOPATHY. Singapore Med J., 31; pp. 463-465

No Published

(Doc II A3) Niemann, L. et al. 2002 TOXICITY TO MAMMALS INCLUDING HUMANS In: Schmutterer, H. (ed.): THE NEEM TREE. Neem foundation, Mumbai/India Report no.: not applicable GLP: unknown Published: yes

No Published

(Doc II A3) Schmutterer, H. (ed.)

2002 THE NEEM TREE. Neem foundation, Mumbai/India Report no.: not applicable GLP: unknown Published: yes

No Published

(Doc II A3) Sri Ranganathan, S. et al.

2005 KOHOMBA OIL INDUCED ENCEPHALOPATHY : LESSONS IN PRESCRIBING TRADITIONAL MEDICINES. Sri Lanka Journal of Child Health, 34, Pp. 94-95

No Published

(Doc II A3) Donghade R.K. et al.

2008 NEEM OIL POISONING. Indian Pediatrics, 45, pp. 56-57

No Published

(Doc II A3) Senanayake M.P. et al.

2009 MARGOSA (KOHOMBA) OIL INDUCED TOXIC ENCEPHALOPATHY FOLLOWING HOME REMEDY FOR INTESTINAL WORMS. Ceylon Med. J., 54/4: p. 140

(Doc II A3) Bhaskar M.V. et al.

2010 MR IMAGING FINDINGS OF NEEM OIL POISONING AJNR, 31, pp. E60 – E61

No Published

(Doc II A3) Iyyadurai R. et. al.

2010 AZADIRACHTIN POISONING: A CASE REPORT. Clinical Toxicology, 48/8, pp. 857-858

No Published

A 6.12.2/03 Pfau, W. 2006 Data bank search - Trifolio-M GmbH Report-no. not applicable GLP/GEP: no Published: no

no TRF

(Doc II A4, B8, C13)

European Chemicals Bureau

2003 Technical Guidance Documents in Support of Directive 93/87/EEC on Risk Assessment for New Notified Substances and The Commission Regulation (EC) 1488/94 on Risk Assessment for Existing Substances, Part II

No Published

(Doc II A4)

Ley, S. 2007 ANGEWANDTE CHEMIE, 119, 40, 7773-7776 No Published


76

A 7.1.1.1.1/01* Troß, R. 1996a Hydrolytic Stability of NeemAzal [German Original] Trifolio-M GmbH, D-35633 Lahnau Trifolio-M GmbH Report-no. TM 1195.15 GLP: yes, GEP: yes Published: no

no TRF

A 7.1.1.1.1/02 Troß, R. 1997 HYDROLYSIS OF AZADIRACHTIN A AS A FUNCTION OF PH-VALUE AND THE TEMPERATURE - EXTRAPOLATION OF HYDROLYTIC DATA AT 30 AND 40 °C FROM THE INVESTIGATION "HYDROLYSIC STABILITY OF NEEMAZAL (TM 1195.15) ON THE TEMPERATURE RANGE OF 18-22°C [GERMAN ORIGINAL] Trifolio-M GmbH, D-35633 Lahnau Trifolio-M GmbH Report-no. LP 97.04 GLP/GEP: no Published: no

no TRF

A 7.1.1.1.2/01* Werle, H. 1995 Direct Phototransformation Study (Quantum Yield) in purified water, OECD Draft Test Guideline, part A, "Direct Phototransformation" NeemAzal BioChem GmbH, Karlsruhe, Germany Trifolio-M GmbH Report-no. 95 50 40 827 B GLP: yes, GEP: yes Published: no

no TRF

A 7.1.1.1.2/02* Werle, H. 1999 Estimation of the Environmental Half Life Time of NeemAzal in the Surface Layer of Aqueous Systems BioChem GmbH, Karlsruhe, Germany Trifolio-M GmbH Report-no. 995040819 GLP: yes, GEP: yes Published: no

no TRF

A 7.1.1.2.1/01* Lenz, G. 1995 Testing the Ready Biodegradability of NeemAzal-T/S in Water by the "Closed Bottle Test" according to OECD Guideline 301 D BioChem GmbH, Karlsruhe, Germany Trifolio-M GmbH Report-no. LP 99.02, 94 50 41 389 GLP: yes Published: no

no TRF

A 7.1.1.2.1/02* Werle, H. 1998 Determination of Ready Biodegradability of NeemAzal "Closed Bottle Test" - according to OECD Guideline 301 D BioChem GmbH, Karlsruhe, Germany Trifolio-M GmbH Report-no. TM0398.01, 97 50 40 787 GLP: yes Published: no

no TRF


77

A 7.1.1.2.1/03* Hund, K. 1998a Biodegradation of NeemAzal: Manometric Respirometry Test Fraunhofer Institut, Schmallenberg-Grafschaft, Germany Trifolio-M GmbH Report-no. LP 99.02, TRF-001/3-15 GLP: yes Published: no

no TRF

A 7.1.1.2.1/04* Hund, K. 1999a Biodegradation of NeemAzal: Manometric Respirometry Test Fraunhofer Institut, Schmallenberg-Grafschaft, Germany Trifolio-M GmbH Report-no. TRF-001/3-15/1 GLP: yes Published: no

no TRF

A 7.1.1.2.1/05* Hund, K. 1999b Biodegradation of Azadirachtin A: Manometric Respirometry Test Fraunhofer Institut, Schmallenberg-Grafschaft, Germany Trifolio-M GmbH Report-no. TM995.10, TRF-003/3-15 GLP: yes Published: no

no TRF

A 7.1.1.2.2/01 Hund, K. 1998a Biodegradation of NeemAzal: Manometric Respirometry Test Fraunhofer Institut, Schmallenberg-Grafschaft, Germany Trifolio-M GmbH Report-no. LP 99.02, TRF-001/3-15 GLP: yes Published: no See also Doc 7.1.1.2.1/03

yes TRF

A 7.1.2.2.1/01 Szeto, A.Y., Wan, M.T.

1996 Hydrolysis of Azadirachtin in Buffered and Natural Waters not applicable Journal of Agricultural and Food Chemistry, Volume 44, No. 4, 1996, pp.1160-1163 Report-no. not applicable GLP/GEP: no Published: yes

no -

A 7.1.2.2.1/02 Sundaram, K.M.S., Sloane, L., Curry J.

1995 Kinetics of Azadirachtin Hydrolysis in Model Aquatic Systems by High-Performance liquid chromatography not applicable Journal of Liquid Chromatography, Volume 18, No. 2, 1995, pp. 363-376 Report-no. not applicable GLP/GEP: no Published: yes

no -


78

A 7.1.2.2.2/01 Szeto, A.Y., Wan, M.T.

1996 Hydrolysis of Azadirachtin in Buffered and Natural Waters not applicable Journal of Agricultural and Food Chemistry, Volume 44, No. 4, 1996, pp.1160-1163 Report-no. not applicable GLP/GEP: no Published: yes Cited in Justification Doc IIIA 7.1.2.2.1/01

no -

A 7.1.2.2.2/02 Sundaram, K.M.S., Sloane, L., Curry J.

1995 Kinetics of Azadirachtin Hydrolysis in Model Aquatic Systems by High-Performance liquid chromatography not applicable Journal of Liquid Chromatography, Volume 18, No. 2, 1995, pp. 363-376 Report-no. not applicable GLP/GEP: no Published: yes Cited in Justification Doc IIIA 7.1.2.2.1/02

no -

A 7.1.2.2.2/03 Sundaram, M.A.K., Sundaram, A., Curry, J., Sloane, L.

1997 Formulation Selection, and Investigation of Azadirachtin-A Persistence in Some Terrestrial and Aquatic Compenents of a Forest Environment not applicable Pesticide Science, Volume 51, Issue 1, 1997, pp. 74-90 Report-no. not applicable GLP/GEP: no Published: yes Cited in Justification Doc IIIA 7.1.2.2.2

no -

A 7.1.2.2.2/04 Thompson, D.G., Kreutzweiser, D.P., Staznik, B., Chartrand, D., Capell, S.

2002 Fate and Persistence of Azadirachtin A following applications to mesocosms in a small forest lake - Bulletin of Environmental Contamination and Toxicology, Vol. 69, No. 2, 2002, pp. 250 to 256 Report-no. Not applicable, not applicable GLP/GEP: no Published: yes Cited in Doc IIIA 7.1.2.2.2/01

no -

(Doc II C13) EC 2006 REACH-VO Regulation (EC) No 1907/2006 of the European Parliament and of the Council of 18 December 2006 concerning the registration, authorisation and restriction of chemicals (REACH) establishing a European Chemicals Agency amending Directive 1999/45/EC and repealing Council Regulation (EEC) No 793/93 and Commission Regulation (EC) No 1488/94 as well as Council Directive 76/769/EEC, Commission Directives 91/155/EEC, 93/105/EC and 2000/21/EC

A 7.1.3/01* Troß, R. 1996b Adsorption and desorption of NeemAzal in the soil Trifolio-M GmbH, D-35633 Lahnau Trifolio-M GmbH Report-no. TM 995.12 GLP: yes, GEP: yes Published: no

no TRF


79

A 7.2.1/01 Kleeberg, H. 1999 Degradation of Azadirachtins and Identification of Metabolites in Soil Trifolio-M GmbH, D-35633 Lahnau Trifolio-M GmbH Report-no. LP 99.02 GLP/GEP: no Published: no

no TRF

A 7.2.2.1/01* Troß, R. 1999 Fate of NeemAzal in the soil - degradation study in three field-moist soils Trifolio-M GmbH, D-35633 Lahnau Trifolio-M GmbH Report-no. TM 0398.01 GLP: yes, GEP: yes Published: no

no TRF

A 7.2.2.1/02* Troß, R. 1995a Degradation of NeemAzal in the soil Trifolio-M GmbH, D-35633 Lahnau Trifolio-M GmbH Report-no. TM 995.10 GLP: yes Published: no

no TRF

A 7.2.2.4/01 Sundaram, K.M.S., Curry, J.

1996 Effect of some UV Light Absorbers on the Photostabilization of Azadirachtin, A Neem-Based Biopesticide not applicable, 1996, pp. 649-659 Report-no. not applicable GLP/GEP: no Published: yes

no -

(Doc II A4) EC 1991 Council Directive 91/414/EEC concerning the placing of plant protection products on the market

No Published

A 7.2.3.1/01 Troß, R.* 1996b Adsorption and desorption of NeemAzal in the soil Trifolio-M GmbH, D-35633 Lahnau Trifolio-M GmbH Report-no. TM 995.12 GLP: yes, GEP: yes Published: no See also Doc IIIA 7.1.3/01

no TRF

A 7.2.3.2/01* Troß, R. 1995b Leaching behaviour of NeemAzal-T/S in the soil Trifolio-M GmbH, D-35633 Lahnau Trifolio-M GmbH Report-no. TM 995.11 GLP: yes, GEP: yes Published: no

no TRF

A 7.3.1/01* Müller, M. 1999 Estimation of the Atmospheric Half-Life of Azadirachtin A Fraunhofer Institut, Schmallenberg-Grafschaft, Germany Trifolio-M GmbH Report-no. not stated, not applicable GLP/GEP: no Published: no

no TRF

A 7.4.1.1/01* 1996 NeemAzal-T/S - Acute Toxicity Fish Trifolio-M GmbH Report-no. 94 50 41 389 C GLP: yes, GEP: yes Published: no

yes TRF


80

A 7.4.1.1/02 1996 Acute Toxicity Study of NeemAzal-T/S to Fresh Water Fish (Common Carp) Trifolio-M GmbH Report-no. 885/JRF/TOX/96 GLP: yes, GEP: yes Published: no

yes TRF

A 7.4.1.2/01* Schmitz, A. 1999c Effects of NeemAzal on Daphnia magna, Acute Immobilization Test and Reproduction Test Fraunhofer Institut, Schmallenberg-Grafschaft, Germany Trifolio-M GmbH Report-no. TRF-001/4-21 GLP: yes, GEP: yes Published: no

yes TRF

A 7.4.1.3/01* Wenzel, A. 2002 Alga, Growth Inhibition Test Effects of NeemAzal on the growth of Scenedesmus subspicatus Fraunhofer Institut, Schmallenberg-Grafschaft, Germany Trifolio-M GmbH Report-no. TRF-001/4-30 GLP: yes, GEP: yes Published: no

yes TRF

A 7.4.1.4/01* Hund, K. 1998b Effect of NeemAzal on the Respiration Activity of activated Sludge Fraunhofer Institut, Schmallenberg-Grafschaft, Germany Trifolio-M GmbH Report-no. TRF-001/3-09 GLP: yes, GEP: yes Published: no

yes TRF

A 7.4.3.2/01* . 2000 Effects of NeemAzal on Fish: Full Life Cycle Test with the Zebra Fish, Danio rerio Trifolio-M GmbH Report-no. TRF-001/4-60 GLP: yes, GEP: yes Published: no

yes TRF

A 7.4.3.2/02* 1999 Effects of NeemAzal-T/S on fish: Prolonged toxicity test (28 d study) with the rainbow trout, Oncorhynchus mykiss; Trifolio-M GmbH Report-no. TRF-002/4-17 GLP: yes Published: no

Yes TRF

A 7.4.3.4/01* Schmitz, A. 1999c Effects of NeemAzal on Daphnia magna, Acute Immobilization Test and Reproduction Test Fraunhofer Institut, Schmallenberg-Grafschaft, Germany Trifolio-M GmbH Report-no. TRF-001/4-21 GLP: yes, GEP: yes Published: no See also Doc IIIA 7.4.1.2/01

yes TRF


81

A7.4.3.4/02* Schmitz, A. 1999d Effects of NeemAzal-T/S on the reproduction of Daphnia magna; Fraunhofer Institut, Schmallenberg-Grafschaft, Germany Trifolio-M GmbH Report-no. TRF-002/4-21 GLP: yes Published: no

yes TRF

A 7.4.3.5.1* Gonsior, G. 2008a Assessment of side effects of Neemazal on the larvae of the midge, chironomus riparius with the laboratory test method, eurofins-GAB GmbH, Germany trifolio-M GmbH Report No. 20071356/01-ASCr GLP: yes Published: no

Yes TRF

A 7.5.2.1* Adelberger, I. 2009 Acute and sublethal toxicity on the predatory mite Hypoaspis (Geolaelaps) aculeifer Canestrini (Acari: laelapidae) using standard soil (lufa 2.2) with freshly applied and aged residues. eurofins-GAB GmbH, Germany trifolio-M GmbH Report No. S09-00977 GLP: yes Published: no

yes TRF

A 7.5.3.1.1/01* 1996a NeemAzal Technical: Acute Oral Toxicity (LD 50) to Bob-white Quail Trifolio-M GmbH Report-no. EIP 21/960383 GLP: yes, GEP: yes Published: no

yes TRF

A 7.5.3.1.2/01* 1996b NeemAzal Technical: Dietary Toxicity (LD 50) to the Bob-white Quail Trifolio-M GmbH Report-no. EIP 22/960382 GLP: yes, GEP: yes Published: no

yes TRF

A 7.5.4.1/01* Klug, T. 2008a NeemAzal®:Toxicity to the green lacewing, Chrysoperla carnea steph., (Neuroptera, Chrysopidae) under extended laboratory conditions – dose response test. eurofins-GAB GmbH, Germany trifolio-M GmbH Report No. 20071356/01-NECc GLP: yes Published: no

Yes TRF

A 7.5.4.1/02 Fifi, A.P. 2007a Effect toxicity evaluation of Neemazal (34 % azadirachtin A) technical product on the coccinella septempunctata l. (Coleoptera, coccinellidae) under extended laboratory conditions. Biotecnologie BT s.r.l., Italy trifolio-M GmbH Report No. BT025/07 GLP: yes Published: no

Yes TRF


82

A 7.5.4.1/03 Mühlen, W., Leymann, B.

1997 Assessment of effects of the plant protection product neemazal-T/S on the bee brood development of the honey bee (apis mellifera l.) – semi-field test Institut für Pflanzenschutz, Saatgutuntersuchung und Bienenkunde, Germany trifolio-M GmbH Report No. BH/LWK/96/2 GLP: no Published: no

Yes TRF

(Doc II C13) EC 1998 The Drinking Water Directive 98/83/EC No Published

* key study


83

Reference list of studies on the biocidal product NeemAzal-T/S

Section No /

Reference No

Author(s) Year Title

Source (where different from company)

Company

Report No.

GLP (where relevant)

(Un)Published

Data

Protectio

n

Claimed

(Yes/No)

Owner

B 3.1/01 Kleeberg, H.

1994 PHYSICO-CHEMICAL PROPERTIES OF THE FORMULATION NEEMAZAL-T/S APPEARANCE, COLOUR AND ODOUR Trifolio-M GmbH, D-35633 Lahnau Trifolio-M GmbH Report-no. LP 94.01 GLP/GEP: no Published: no

yes TRF

B 3.2/01

Battersby, R.V. 2002a DOCUMENT K, ANNEX II POINT 2.2.1, EXPLOSIVITY NEEMAZAL-T/S EBRC Consulting GmbH, Hannover, Germany Trifolio-M GmbH Report-no. TFM-020505-04 GLP/GEP: no Published: no

yes TRF

B 3.3/01 Battersby, R.V.

2002b DOCUMENT K, ANNEX III POINT 2.2.2, OXIDISING PROPERTIES NEEMAZAL-T/S EBRC Consulting GmbH, Hannover, Germany Trifolio-M GmbH Report-no. TFM-020505-05 GLP/GEP: no Published: no

yes TRF

B 3.4/01 Werle, H.

1995a FLASH POINT, EC DIRECTIVE 92/69/EEC,A.9. NEEMAZAL-T/S BioChem GmbH, Karlsruhe, Germany Trifolio-M GmbH Report-no. 94 50 40 389 B GLP: yes, GEP: yes Published: no

yes TRF

B 3.4/02 Franke, J.

2005 AUTO-FLAMMABILITY (DETERMINATION OF THE TEMPERATURE OF SELF-IGNITION OF VOLATILE LIQUIDS AND OF GASES) A.15. Siemens AG, Prozess-Sicherheit, Frankfurt am Main, Germany Trifolio-M GmbH Report-no. 20050678.01 GLP: yes Published: no

yes TRF

B 3.5/01 Troß, R.

1995a DETERMINATION OF THE PH - VALUE OF NEEMAZAL- T/S [GERMAN ORIGINAL] Trifolio-M GmbH, D-35633 Lahnau Trifolio-M GmbH Report-no. LP 95.3 GLP/GEP: no Published: no

yes TRF


84

B 3.6/01 Troß, R.

1995b DETERMINATION OF DENSITY OF THE PLANT PROTECTION PRODUCT NEEMAZAL-T/S [GERMAN ORIGINAL] Trifolio-M GmbH, D-35633 Lahnau Trifolio-M GmbH Report-no. TM 895.6 GLP: yes, GEP: yes Published: no

yes TRF

B 3.7/01 Hummel E., Troß R.

1996 STORAGE STABILITY ON NEEMAZAL-T/S: EFFICACY IN THE BIOASSAY [GERMAN ORIGINAL] Trifolio-M GmbH, D-35633 Lahnau Trifolio-M GmbH Report-no. LP 95.8 GLP/GEP: no Published: no

yes TRF

B 3.7/02 Troß, R.

1995c STORAGE STABILITY OF NEEMAZAL-T/S [GERMAN ORIGINAL] Trifolio-M GmbH, D-35633 Lahnau Trifolio-M GmbH Report-no. LP 95.1 GLP/GEP: no Published: no

yes TRF

B 3.7/03 Ruch B.

2005 NEEMAZAL-T/S: LOW TEMPERATURE STABILITY Trifolio-M GmbH, D-35633 Lahnau Trifolio-M GmbH Report-no. TM 0905.04 GLP: yes Published: no

yes TRF

B 3.7/04 Troß, R.

1996 STORAGE STABILITY OF NEEMAZAL-T/S (REAL TIME TEST) [GERMAN ORIGINAL] Trifolio-M GmbH, D-35633 Lahnau Trifolio-M GmbH Report-no. LP 96.1 GLP/GEP: no Published: no

yes TRF

B 3.8/01 Ruch, B.

2005 DETERMINATION OF THE EMULSION CHARACTERISTICS AND RE-EMULSIFICATION PROPERTIES OF NEEMAZAL-T/S Trifolio-M GmbH, D-35633 Lahnau Trifolio-M GmbH Report-no. TM 0805.01 GLP: yes Published: no

yes TRF

B 3.8/02 Troß, R.

1995d DETERMINATION OF THE FOAM STABILITY OF NEEMAZAL-T/S [GERMAN ORIGINAL] Trifolio-M GmbH, D-35633 Lahnau Trifolio-M GmbH Report-no. LP 95.6 GLP/GEP: no Published: no

yes TRF


85

B 3.10.1/01 Walter, D. 2004 SURFACE TENSION OF THE FORMULATION NEEMAZAL-T/S GAB Biotechn. GmbH & GAB Analytik GmbH, Niefern-Öschelbronn Trifolio-M GmbH Report-no. 20031200/01-PCST GLP: yes Published: no

yes TRF

B 3.10.2/01 Werle, H.

1995b VISCOSITY, OECD-GUIDELINE NO. 114 NEEMAZAL-T/S BioChem GmbH, Karlsruhe, Germany Trifolio-M GmbH Report-no. 94 50 40 389 A GLP: yes, GEP: yes Published: no

yes TRF

B 5.10.2/01 Breuer. M., de Loof, A.

2000 LABORATORY TRIALS WITH NEEMAZAL T/S ON THE ALLERGENIC FOREST PEST THAUMETOPOEA PROCESSIONEA (L.), IN: PRACTICE ORIENTATED RESULTS ON USE AND PRODUCTION OF NEEM-INGREDIENTS AND PHEROMONES VIII Zoological Institute, Kathol. Universiteit Leuven, Belgium Proc of the 8th workshop, Hohensolms, Germany, 2000 Report-no. GLP/GEP: no Published: yes

no -

B 5.10.2/02 Lehman, M., Fieguth, A.

2000 FURTHER RESULTS IN USING NEEMAZAL T/S AGAINST OAK PROCESSION MOTH (THAUMETOPOEA PROCESSIONEA LINNE), IN: PRACTICE ORIENTATED RESULTS ON USE AND PRODUCTION OF NEEM-INGREDIENTS AND PHEROMONES VIII Pflanzenschutzdienst LA, F. a.d.O. Germany Proc of the 8th workshop, Hohensolms, Germany Report-no. GLP/GEP: no Published: yes

no -

B 6.1.1/01 1997a NEEMAZAL-FORMULATION: ACUTE ORAL TOXICITY TO THE RAT Trifolio-M GmbH Report-no. EIP 15/950801/AC GLP: yes, GEP: yes Published: no

yes TRF

B 6.1.2/01 1997b NEEMAZAL FORMULATION: ACUTE DERMAL TOXICITY TO THE RAT Trifolio-M GmbH Report-no. EIP 16/950802/AC GLP: yes, GEP: yes Published: no

yes TRF


86

B 6.1.3/01 1997 NEEMAZAL FORMULATION: ACUTE INHALATION TOXICITY TO RATS, 4 HOUR EXPOSURE Trifolio-M GmbH Report-no. EIP 14/951591 GLP: yes, GEP: yes Published: no

yes TRF

B 6.2.1/01 1996a NEEMAZAL FORMULATION: SKIN IRRITATION TO THE RABBIT Trifolio-M GmbH Report-no. EIP 17/950824/SE GLP: yes, GEP: yes Published: no

yes TRF

B 6.2.2/01 1996b NEEMAZAL FORMULATION: EYE IRRITATION TO THE RABBIT Trifolio-M GmbH Report-no. EIP 18/950825/SE GLP: yes, GEP: yes Published: no

yes TRF

B 6.3/01 1998 SKIN SENSITISATION STUDY ACCORDING TO MAGNUSSON & KLIGMAN Trifolio-M GmbH Report-no. 981042830 GLP: yes Published: no

yes TRF

B 6.3/02 1997 NEEMAZAL FORMULATION SKIN SENSITISATION IN THE GUINEA-PIG Trifolio-M GmbH Report-no. EIP 19/951048/SS GLP: yes Published: no

yes TRF

B 6.4/01 2005 STUDIES ON THE TRANSDERMAL PENETRATION AND RESORPTION OF NEEMAZAL SHAMPOO AND NEEMAZAL T/S [GERMAN ORIGINAL] not applicable Trifolio-M GmbH Report-no. not applicable GLP/GEP: no Published: no

yes TRF

B 6.4/02 Kleeberg, H. 2007 STATEMENT TO: STUDIES ON THE TRANSDERMAL PENETRATION AND RESORPTION OF NEEMAZAL SHAMPOO AND NEEMAZAL T/S [GERMAN ORIGINAL] Trifolio-M GmbH, D-35633 Lahnau Trifolio-M GmbH Report-no. not applicable GLP/GEP: no Published: no

yes TRF


87

B 6.4/03 Kietzmann, M., Löscher, W., Arens, D., Maaß, P., Lubach, D.

1993 THE ISOLATE PERFUSED BOVINE UDDER AS AN IN VITRO MODEL OF PERCUTANEOUS DRUG ABSORPTION SKIN VIABILITY AND PERCUTANEOUS ABSORPTION OF DEXAMETHASONE, BENZOYL PEROXIDE, AND ETOFENAMTE not stated Journal Pharmacol. Toxicol. Methods, 30, No. 2, 75 - 84 Report-no. not stated GLP/GEP: no Published: yes

no -

B 6.4/04 Pfau, W. 2007 STATEMENT ON DERMAL ABSORPTION OF AZADIRACHTIN GAB Consulting GmbH, Lamstedt, Germany Trifolio-M GmbH Report-no. 243126-A2-070821-01 GLP/GEP: no Published: no

yes TRF

(Doc II B8) Hoernicke, E. et al.

1998 Details in the instructions for use on the protection of persons carrying out successive work with crops which have been treated with plant protection products. Nachrichtenbl. Deut. Pflanzenschutzd. 50, 267-268; in conjunction with: Krebs et al. (2000): Uniform principals for ensuring health protection for workers when re-entering treated crops following the application of plant protection products. Nachrichtenbl. Deut. Pflanzenschutzd. 52, 5-9 (both in German).

No Published

Doc II B8 Ganzelmeier, H. & Rautmann, D.

2000 Drift, drift reducing sprayers and sprayer testing. Aspects of Applied Biology 57, 1-11.

No Published

Doc II B8 EC

June 2002

Technical Notes for Guidance: Human Exposure to Biocidal Products - Guidance on Exposure Estimation [„Report 2002“ http://ecb.jrc.it/biocides]

No Published

Doc II B8 Warren, N, Marquart, H., Christopher, Y.; Laitinen, J.; Van Hemmen, J.

2006 RISKOFDERM (2003) Deliverable 38: Dermal exposure assessment models for risk assessment Task-based dermal exposure models for regulatory risk assessment, Ann. Occup. Hyg. Vol. 50, 491-503

No Published

Doc II B8 OECD 2003 OECD SERIES ON EMISSION SCENARIO DOCUMENTS, for insecticides, acaricides and products to control other arthropods (PT18) for household and professional uses (5th draft, March 2008)

No Published

Doc II B8 EC 1975 Directive on Waste Disposal 75/442/EEC No Published


88

B 7.8.2/01 Kling, A. 2001 ASSESSMENT OF SIDE EFFECTS OF NEEMAZAL-T/S TO THE HONEY BEE, APIS MELLIFERA L. IN THE LABORATORY GAB Biotechn. GmbH & IFU Umweltanalytik GmbH, Germany Trifolio-M GmbH Report-no. 20011306/01-BLEU GLP: yes, GEP: yes Published: no

yes TRF

B 7.8.3/01 Kühner, C. 1996 NEEMAZAL-T/S: ACUTE TOXICITY TO THE APHID PARASITOID, APHIDIUS RHOPALOSIPHI (HYMENOPTERA, APHIDIIDAE) IN THE LABORATORY GAB Biotechn. GmbH & IFU Umweltanalytik GmbH, Germany Trifolio-M GmbH Report-no. 96164/01-NLAp GLP: yes, GEP: yes Published: no

yes TRF

B 7.8.3/02 Zebitz, C. 1998 RESEARCH PROPOSAL - DEVELOPMENT OF A PRODUCT FOR THE BIOLOGICAL CONTROL OF APHIDS IN ORCHARDS BASED ON RENEWABLE PRIMARY PRODUCTS - FINAL REPORT [GERMAN ORIGINAL] Universität Hohenheim, Germany Trifolio-M GmbH Report-no. AZ 04822 GLP/GEP: no Published: no

yes TRF

B 7.8.3/03 Klenner, Kaboth, G.

1995a TESTING OF THE EFFECTS OF NEEMAZAL-T/S ON LARVAE OF THE LADYBIRD COCCINELLA SEPTEMPUNCTATA L. -LABORATORY TEST- Institut für Pflanzenschutz (IPSAB), D-48147 Münster Trifolio-M GmbH Report-no. ML402 GLP: yes, GEP: yes Published: no

yes TRF

B 7.8.3/04 Klenner, Kaboth, G.

1995b TESTING OF THE EFFECTS OF NEEMAZAL-T/S ON POECILUS CUPREUS L. (COLEOPTERA, CARABIDAE) IN THE LABORATORY Institut für Pflanzenschutz (IPSAB), D-48147 Münster Trifolio-M GmbH Report-no. LL405 GLP: yes, GEP: yes Published: no

yes TRF


89

B 7.8.3/05 Drescher, K. 1995 EFFECT OF A NEEM EXTRACT (MELIACEAE; AZADIRACHTA INDICA A. JUSS) ON VITALITY AND FECUNDITY OF THE PREDATORY BUG ORIUS MAJUSCULUS REUTER (HETEROPTERA; ANTHOCORIDAE) [GERMAN ORIGINAL] Universität Bonn, Germany Trifolio-M GmbH Report-no. not applicable GLP/GEP: no Published: no

yes TRF

B 7.8.3/06 Kühner, C. 1997a NEEMAZAL-T/S: DETERMINATION OF SIDE-EFFECTS ON THE APHID PARASITOID, APHIDIUS SPP. (HYMENOPTERA, APHIDIIDAE) USING AN EXTENDED LABORATORY TEST GAB Biotechn. GmbH & IFU Umweltanalytik GmbH, Germany Trifolio-M GmbH Report-no. 96164/01-NEAp GLP: yes, GEP: yes Published: no

yes TRF

B 7.8.3/07 Kühner, C. 1997b NEEMAZAL-T/S: ASSESSMENT OF SIDE EFFECTS ON LARVAE OF THE HOVERFLY, EPISYRPHUS BALTEATUS DEG. (DIPTERA, SYRPHIDAE) UNDER SEMI-FIELD CONDITIONS GAB Biotechn. GmbH & IFU Umweltanalytik GmbH, Germany Trifolio-M GmbH Report-no. 96164/02-NHEb GLP: yes, GEP: yes Published: no

yes TRF

B 7.8.3/08 Hermann, P. 2001 SIDE EFFECTS OF THE PLANT-DERIVED INSECTICIDE NEEMAZAL-T/S TO NON-TARGET ARTHROPODS RELEVANT IN ORCHARDS [GERMAN ORIGINAL] Universität Hohenheim, Germany Trifolio-M GmbH Report-no. not applicable GLP/GEP: no Published: no

yes TRF

B 7.8.3/09 Rogler, T. 1994 EFFECT OF NEEM FORMULATIONS ON APHIDS, WINTER MOTHS AND PREDATORY MITES IN AN APPLE ORCHARD IN COMPARISON TO PIRIMOR AND NEUDOSAN [GERMAN ORIGINAL] Fachhochschule Wiesbaden, BRD Trifolio-M GmbH Report-no. not applicable GLP/GEP: no Published: no

yes TRF


90

B 7.8.3/10 Ruess, F. 1996 FIELD STUDY FOR TESTING OF NEEMAZAL-T/S TO PREDATORY MITES ((TYPHLODROMUS PYRI) IN VINEYARDS [GERMAN ORIGINAL] Staatliche Lehr- und Versuchsanstalt, D-74185 Weinsberg Trifolio-M GmbH Report-no. 96 02 TRI 1 GLP: yes, GEP: yes Published: no

yes TRF

B 7.8.3/11 Klug, T. 2008a NEEMAZAL-T/S: TOXICITY TO THE GREEN LACEWING CHRYSOPERLA CARNEA STEPH. (NEUROPTERA, CHRYSOPIDAE) UNDER EXTENDED LABORATROY CONDITIONS - DOSE-RESPONSE TEST Eurofins-GAB GmbH, Germany Trifolio-M GmbH Report-no. 20071355/01-NECc GLP: yes, Published: no

Yes TRF

B 7.8.3/12 Klug, T. 2008b NEEMAZAL-T/S: TOXICITY TO THE GREEN LACEWING CHRYSOPERLA CARNEA STEPH. (NEUROPTERA, CHRYSOPIDAE) USING AN EXTENDED LABORATROY TEST WITH FRESHLY APPLIED AND AGED RESIDUES Eurofins-GAB GmbH, Germany Trifolio-M GmbH Report-no. S08-01908 GLP: yes, Published: no

Yes TRF

B 7.8.3/13 Klug, T. 2008c NEEMAZAL-T/S: TOXICITY TO THE GREEN LACEWING CHRYSOPERLA CARNEA STEPH. (NEUROPTERA, CHRYSOPIDAE) USING AN EXTENDED LABORATROY TEST WITH FRESHLY APPLIED AND AGED RESIDUES Eurofins-GAB GmbH, Germany Trifolio-M GmbH Report-no. S08-02528 GLP: yes, Published: no

Yes TRF

B 7.8.3/14 Klug, T. 2009 NEEMAZAL-T/S: TOXICITY TO THE GREEN LACEWING CHRYSOPERLA CARNEA STEPH. (NEUROPTERA, CHRYSOPIDAE) UNDER EXTENDED LABORATROY CONDITIONS - DOSE-RESPONSE TEST Eurofins-GAB GmbH, Germany Trifolio-M GmbH Report-no. BT011/09 GLP: yes, Published: no

Yes TRF


91

B 7.8.3/15 Colli, M. 2009 EFFECTS OF NEEMAZAL-T/S ON THE PLANT DWELLING INSECT COCCINELLA SEPTEMPUNCTATA L. (COLEOPTERA: COCCINELLIDAE) UNDER EXTENDED LABORATORY CONDITION (RATE RESPONSE TEST) Biotecnologie BT s.r.l., Italy Trifolio-M GmbH Report-no. S08-03154 GLP: yes, Published: no

Yes TRF

B 7.8.4/01 Hänisch, Bathelt, A.

1995 TESTING OF THE EFFECTS OF NEEMAZAL-T/S ON EARTHWORM EISENIA FETIDA IN THE LABORATORY Institut für Pflanzenschutz (IPSAB), D-48147 Münster Trifolio-M GmbH Report-no. RL 406 GLP: yes, GEP: yes Published: no

yes TRF

B 7.8.5/01 Kölzer, U. 2005 ASSESSSMENT OF THE SIDE EFFECTS OF NEEMAZAL-T/S ON THE ACTIVITY OF THE SOIL MICROFLORA GAB Biotechn. GmbH & IFU Umweltanalytik GmbH, Germany Trifolio-M GmbH Report-no. 20051121/01-ABMF GLP: yes, GEP: yes Published: no

yes TRF

B 7.8.6/01 Martinez, E.S. 2002 VEGETATIVE VIGOUR LIMIT TEST FOR NON TARGET PLANTS, FOLOWING SINGLE RATE APPLICATION OF NEEMAZAL-T/S IN GREENHOUSE UNDER CONTROLLED CLIMATIC CONDITIONS IN SPAIN, 2001 GAB Biotechn. GmbH & IFU Umweltanalytik GmbH, Germany Trifolio-M GmbH Report-no. 20017011/S1-FNTP GLP: yes, GEP: yes Published: no

yes TRF