title: Commission Directive 1999/27/EC of 20 April 1999 establishing Community methods of analysis for the determination of amprolium, diclazuril and carbadox in feedingstuffs and amending Directives 71/250/EEC, 73/46/EEC and repealing Directive 74/203/EEC (Text with EEA relevance)

text: L 118/36 EN Official Journal of the European Communities 6. 5. 1999 COMMISSION DIRECTIVE 1999/27/EC of 20 April 1999 establishing Community methods of analysis for the determination of ampro- lium, diclazuril and carbadox in feedingstuffs and amending Directives 71/ 250/EEC, 73/46/EEC and repealing Directive 74/203/EEC (Text with EEA relevance) THE COMMISSION OF THE EUROPEAN COMMUNITIES, Having regard to the Treaty establishing the European Community, Having regard to Council Directive 70/373/EEC of 20 July 1970 on the introduction of Community methods of sampling and analysis for the official control of feeding- stuffs (1), as last amended by the Act of Accession of Austria, Finland and Sweden, and in particular Article 2 thereof, (1) Whereas Directive 70/373/EEC stipulates that offi- cial controls of feedingstuffs for the purpose of checking compliance with the requirements arising under regulations and administrative provisions governing their quality and composition must be carried out using Community methods of sampling and analysis; the laws, (2) Whereas Council Directive 70/524/EEC of 23 November 1970 concerning additives in feeding- stuffs (2), as last amended by Commission Regula- tion 45/1999 (3) stipulates that the amprolium and diclazuril content must be indicated on the label- ling where these substances are added to premix- tures and feedingstuffs; whereas the authorisation of carbadox for use as a feed additive has been with- drawn by Commission Regulation 2788/98 of 22 December 1998 amending Council Directive 70/ 524/EEC concerning additives in feedingstuffs as regards the withdrawal of authorisation for certain growth promoters (4) and official control of possible illegal use of prohibited substances is necessary; (3) Whereas Community methods of analysis must be established for checking these substances; (4) Whereas the first Commission Directive 71/ 250/EEC of 15 June 1971 establishing Community methods of analysis for the official control of feedingstuffs (5), as last amended by Directive 98/ 54/EC (6) sets out methods of analysis for, inter alia, the determination of mustard oil asnd theo- bromine; whereas the methods described are no longer valid in the light of advances in scientific intended and technical knowledge their for (1) OJ L 170, 3. 8. 1970, p. 2. (2) OJ L 270, 14. 12. 1970, p. 1. (3) OJ L 6, 12. 1. 1999, p. 3. (4) OJ L 347, 23. 12. 1998, p. 31. (5) OJ L 155, 12. 7. 1971, p. 13. (6) OJ L 208, 24. 7. 1998, p. 49. purpose; whereas it delete these methods; is therefore appropriate to 1972 (5) Whereas the fourth Commission Directive 73/ 46/EEC of establishing 5 December Community methods of analysis for the official feedingstuffs (7), as last amended by control of Directive 98/54/EC sets out methods of analysis retinol for, (vitamin A); whereas the method described is no longer valid in the light of advances in scientific and technical knowledge for the intended purpose; whereas it is therefore appropriate to delete the method for retinol; the determination of inter alia, (6) Whereas 1974 25 March sets out methods the fifth Commission Directive 74/ 203/EEC of establishing Community methods of analysis for the official feedingstuffs (8), as last amended by control of Directive 81/680/EEC (9), for analysis for the determination of starch and starch degradation products of high molecular weight in feedingstuffs containing beet cossettes, beet pulp, dried beet tops or leaves, potato pulp, dried yeasts, products rich in inulin or greaves, amprolium, ethopabate, dinitolmide, nicarbazin and menadione (vitamin K3); whereas all the methods described in that Directive are no longer valid in the light of advances in scientific and technical knowledge for their is therefore intended purpose; whereas it appropriate to repeal this Directive; (7) Whereas the measures provided for in this Direc- tive are in accordance with the opinion of the Standing Committee on Feedingstuffs, HAS ADOPTED THIS DIRECTIVE: Article 1 The Member States shall provide that analyses conducted with a view to official controls of the amprolium, dicla- zuril and carbadox content of feedginstuffs and premix- tures are carried out using the methods set out in the Annex hereto. (7) OJ L 83, 30. 3. 1973, p. 21. (8) OJ L 108, 22. 4. 1974, p. 7. (9) OJ L 246, 29. 8. 1981, p. 32. 6. 5. 1999 EN Official Journal of the European Communities L 118/37 Article 2 They shall apply the measures from 1 November 1999. Directive 71/250/EEC is amended as follows. 1. In article 1 the words ‘mustard oil' and ‘theobromine' are deleted. 2. Points 8 and 13 of the Annex are deleted. When Member States adopt these measures, they shall contain a reference to this Directive or shall be accompa- nied by such reference at the time of their official publi- cation. The procedure for such reference shall be adopted by Member States. Article 3 Directive 73/46/EEC is amended as follows. 1. Article 2 is deleted. 2. Annex II is deleted. Article 4 The Directive 74/203/EEC is repealed. Article 6 This Directive shall enter into force on the 20th day following its publication in the Official Journal of the European Communities. Article 7 This Directive is addressed to the Member States. Article 5 Done at Brussels, 20 April 1999. Member States shall bring into force, not later than 31 October 1999, the laws, regulations or administrative provisions necessary to comply with the provisions of this Directive. They shall immediately inform the Commis- sion thereof. For the Commission Franz FISCHLER Member of the Commission L 118/38 EN Official Journal of the European Communities 6. 5. 1999 ANNEX Part A DETERMINATION OF AMPROLIUM 1-[(4-amino-2-propylpyrimidin-5-yl)methyl]-2-methyl-pyridinium chloride hydrochloride 1. Purpose and scope This method is for the determination of amprolium in feedingstuffs and premixtures. The detection limit is 1 mg/kg, the limit of determination is 25 mg/kg. 2. Principle The sample is extracted with a methanol-water mixture. After dilution with the mobile phase and membrane filtration the content of amprolium is determined by cation exchange high performance liquid chromatography (HPLC) using a UV detector. 3. Reagents 3. 1. Methanol. 3. 2. Acetonitrile, HPLC grade. 3. 3. Water, HPLC grade. 3. 4. Sodium dihydrogen phosphate solution, c = 0,1 mol/l Dissolve 13. 80 g of sodium dihydrogen phosphate monohydrate in water (3. 3) in a 1000 ml graduated flask, make up to the mark with water (3. 3) and mix. 3. 5. Sodium perchlorate solution, c = 1,6 mol/l Dissolve 224,74 g of sodium perchlorate monohydrate in water (3. 3) in a 1000 ml graduated flask, make up to the mark with water (3. 3) and mix. 3. 6. Mobile phase for HPLC (see observation 9. 1). Mixture of acetonitrile (3. 2), sodium dihydrogen phosphate solution (3. 4) and sodium perchlorate solution (3. 5), 450+450+100 (v+v+v). Prior to use filter through a 0. 22 lm membrane filter (4. 3) and degas the solution (e. g. in the ultrasonic bath (4. 4) for at least 15 minutes). 3. 7. Standard substance: pure amprolium, l-[(4-amino-2-propylpyrimidin-5-yl)methyl]-2-mehyl-pyridinium chloride hydrochloride, E 750 (see 9. 2). 3. 7. 1. Amprolium stock standard solution, 500 lg/ml Weigh to the nearest 0,1 mg, 50 mg of amprolium (3. 7) in a 100 ml graduated flask, dissolve in 80 ml methanol (3. 1) and place the flask for 10 min in an ultrasonic bath (4. 4). After ultrasonic treatment bring the solution to room temperature, make up to the mark with water and mix. At a temperature of @ 4 (cid:176)C the solution is stable for one month. 3. 7. 2. Amprolium intermediate standard solution, 50 lg/ml Pipette 5 ml of the stock standard solution (3. 7. 1) into a 50 ml graduated flask, make up to the mark with the extraction solvent (3. 8) and mix. At a temperature of @ 4 (cid:176)C the solution is stable for one month. 3. 7. 3. Calibration solutions Transfer 0,5, 1 and 2 ml of the intermediate standard solution (3. 7. 2). into a series of 50 ml graduated flasks. Make up to the mark with the mobile phase (3. 6) and mix. These solutions correspond to 0,5, 1 and 2 lg of amprolium per ml respectively. These solutions must be prepared freshly before use. 6. 5. 1999 EN Official Journal of the European Communities L 118/39 3. 8. Extraction solvent Methanol (3. 1)-water mixture 2+1 (v+v). 4. Apparatus 4. 1. HPLC equipment with injection system, suitable for injection volumes of 100 ll. 4. 1. 1. Liquid chromatographic column 125 mm x 4 mm, cation exchange Nucleosil 10 SA, 10 lm packing, or equivalent 4. 1. 2. UV detector with variable wavelength adjustment or diode array detector. 4. 2. Membrane filter, PTFE material, 0,45 lm. 4. 3. Membrane filter, 0,22 lm. 4. 4. Ultrasonic bath. 4. 5. Mechanical shaker or magnetic stirrer. 5. Procedure 5. 1. General 5. 1. 1. Blank feed For the performance of the recovery test (5. 1. 2) a blank feed should be analysed to check that neither amprolium nor interfering substances are present. The blank feed should be similar in type to that of the sample and amprolium or interfering substances should not be detected. 5. 1. 2. Recovery test A recovery test should be carried out by analysing the blank feed which has been fortified by addition of a quantity of amprolium, similar to that present in the sample. To fortify at a level of 100 mg/kg, transfer 10 ml of the stock standard solution (3. 7. 1) to a 250 ml conical flask and evaporate the solution to approximately 0,5 ml. Add 50 g of the blank feed, mix thoroughly and leave for 10 minutes mixing again several times before proceeding with the extraction step (5. 2). Alternatively, if a blank feed similar in type to that of the sample is not available (see 5. 1. 1), a recovery test can be performed by means of the standard addition method. In this case, the sample to be analysed is fortified with a quantity of amprolium similar to that already present in the sample. This sample is analysed together with the unfortified sample and the recovery can be calculated by subtraction. 5. 2. Extraction 5. 2. 1. Premixtures (content : 1 % amprolium) and feedingstuffs Weigh to the nearest 0,01 g, 5 to 40 g of the sample depending on the amprolium content into a 500 ml conical flask and add 200 ml extraction solvent (3. 8). Place the flask in the ultrasonic bath (4. 4) and leave for 15 minutes. Remove the flask from the ultrasonic bath and shake it for 1 hour on the shaker or stir on the magnetic stirrer (4. 5). Dilute an aliquot of the extract with the mobile phase (3. 6) to an amprolium content of 0,5 to 2 lg/ml and mix (see observation 9. 3). Filter 5 to 10 ml of this diluted solution on a membrane filter (4. 2). Proceed to the HPLC determination (5. 3). 5. 2. 2. Premixtures (content A l % amprolium) Weigh to the nearest 0,001 g, 1 to 4 g of the premixture depending on the amprolium content into a 500 ml conical flask and add 200 ml extraction solvent (3. 8). Place the flask in the ultrasonic bath (4. 4) and leave for l5 minutes. Remove the flask from the ultrasonic bath and shake it for 1 hour on the shaker or stir on the magnetic stirrer (4. 5). Dilute an aliquot of the extract with the mobile phase (3. 6) to an amprolium content of 0,5 to 2 lg/ml and mix. Filter 5 to 10 ml of this diluted solution on a membrane filter (4. 2). Proceed to the HPLC determination (5. 3). 5. 3. HPLC determination L 118/40 EN Official Journal of the European Communities 6. 5. 1999 5. 3. 1. Parameters: The following conditions are offered for guidance, other conditions may be used provided that they give equivalent results. Liquid chromatographic column (4. 1. 1): 125 mm · 4 mm, cation exchange Nucleosil 10 SA, 10 lm packing, or equivalent. Mobile phase (3. 6): Mixture of acetonitrile (3. 2), sodium dihydrogen phosphate solution (3. 4) and sodium perchlorate solution (3. 5), 450+450+100 (v+v+v). Flow rate: 0,7 to 1 ml/min. Detection wavelength: Injection volume: 264 nm. 100 ll. Check the stability of the chromatographic system, injecting several times the calibration solution (3. 7. 3) containing 1,0 lg/ml, until constant peak heights and retention times are achieved. 5. 3. 2. Calibration graph Inject each calibration solution (3. 7. 3) several times and determine the mean peak heights (areas) for each concentration. Plot a calibration graph using the mean peak heights (areas) of the calibration solutions as the ordinates and the corresponding concentrations in lg/ml as the abscissae. 5. 3. 3. Sample solution Inject the sample extract (5. 2) several times using the same volume as taken for the calibration solutions and determine the mean peak height (area) of the amprolium peaks. 6. Calculation of the results From the mean height (area) of the amprolium peaks of the sample solution determine the concentra- tion of the sample solution in lg/ml by reference to the calibration graph (5. 3. 2). The amprolium content w in mg/kg of the sample is given by the following formula: w = V·b·f m [mg/kg] in which: V = volume of the extraction solvent (3. 8) in ml according to 5. 2 (i. e. 200 ml) b = amprolium concentration of the sample extract (5. 2) in lg/ml f = dilution factor according to 5. 2 m = mass of the test portion in g 7. Validation of the results 7. 1. Identity The identity of the analyte can be confirmed by co-chromatography, or by using a diode-array detector by which the spectra of the sample extract (5. 2) and the calibration solution (3. 7. 3) containing 2,0 lg/ml are compared. 7. 1. 1. Co-chromatography A sample extract (5. 2) is fortified by addition of an appropriate amount of calibration solution (3. 7. 3). The amount of added amprolium should be similar to the amount of amprolium found in the sample extract. Only the height of the amprolium peak should be enhanced after taking into account both the amount added and the dilution of the extract. The peak width, at half of its height, must be within 5 10 % of the original width of the amprolium peak of the unfortified sample extract. 6. 5. 1999 EN Official Journal of the European Communities L 118/41 7. 1. 2. Diode array detection The results are evaluated according to the following criteria: (a) The wavelength of maximum absorption of the sample and of the standard spectra, recorded at the peak apex on the chromatogram, must be the same within a margin determined by the resolving power of the detection system. For diode-array detection this is typically within 5 2 nm. (b) Between 210 and 320 nm, the sample and standard spectra recorded at the peak apex of the chromatogram, must not be different for those parts of the spectrum within the range 10-100 % of relative absorbance. This criterion is met when the same maxima are present and at no observed point the deviation between the two spectra exceeds 15 % of the absorbance of the standard analyte. (c) Between 210 and 320 nm, the spectra of the upslope, apex and downslope of the peak produced by the sample extract must not be different from each other for those parts of the spectrum within the range 10-100 % of relative absorbance. This criterion is met when the same maxima are present and when at all observed points the deviation between the spectra does not exceed 15 % of the absorbance of the spectrum of the peak apex. If one of these criteria is not met, the presence of the analyte has not been confirmed. 7. 2. Repeatability The difference between the results of two parallel determinations carried out on the same sample must not exceed (cid:190) 15 % relative to the higher value for amprolium contents from 25 mg/kg to 500 mg/kg (cid:190) 75 mg/kg for amprolium contents between 500 mg/kg and 1000 mg/kg (cid:190) 7,5 % relative to the higher value for amprolium contents of more than 1000 mg/kg 7. 3. Recovery For a fortified (blank) sample the recovery should be at least 90 %. 8. Results of a collaborative study A collaborative study was arranged in which three poultry feeds (sample 1-3), one mineral feed (sample 4) and one premix (sample 5) were analysed. The results are given in the following table Sample 1 (blank feed) Sample 2 Sample 3 Sample 4 Sample 5 14 56 (cid:190) (cid:190) (cid:190) (cid:190) (cid:190) (cid:190) 14 56 45,5 2,26 4,95 2,95 6,47 50 14 56 188 3,57 1,90 11,8 6,27 200 14 56 5 129 178 3,46 266 5,19 15 60 25 140 550 2,20 760 3,00 5 000 25 000 L n mean [mg/kg] [mg/kg] s r CV [%] r sR [mg/kg] [%] CV R Nominal content [mg/kg] L: n: number of laboratories. number of single values. s : r CV s : R standard deviation of repeatability. r: coefficient of variation of repeatability. standard deviation of reproducibility. CVR : coefficient of variation of reproducibility. L 118/42 EN Official Journal of the European Communities 6. 5. 1999 9. Observations 9. 1. 9. 2. If the sample contains thiamine, the thiamine peak in the chromatogram appears shortly before the amprolium peak. Following this method amprolium and thiamine must be separated. If the ampro- lium and thiamine are not separated by the column (4. 1. 1) used in this method, replace up to 50 % of the acetonitrile portion of the mobile phase (3. 6) by methanol. According to the British Pharmacopoeia, the spectrum of an amprolium solution (c = 0,02 mol/l) in hydrochloric acid (c = 0,1 mol/l) shows maxima at 246 nm and 262 nm. The absorbance shall amount to 0,84 at 246 nm and 0,80 at 262 nm. 9. 3. The extract must always be diluted with the mobile phase, because otherwise the retention time of the amprolium peak may shift significantly, due to changes in the ionic strength. PART B DETERMINATION OF DICLAZURIL (+)-4-chlorphenyl [2,6-dichloro-4-(2,3,4,5-tetrahydro-3,5-dioxo-1,2,4-triazin-2-yl) phenyl] acetonitrile. 1. Purpose and scope The method is for the determination of diclazuril in feedingstuffs and premixtures. The limit of detection is 0,1 mg/kg, the limit of determination is 0,5 mg/kg. 2. Principle After addition of an internal standard, the sample is extracted with acidified methanol. For feeding- stuffs, an aliquot of the extract is purified on a C18 solid phase extraction cartridge. Diclazuril is eluted from the cartridge with a mixture of acidified methanol and water. After evaporation, the residue is dissolved in DMF/water. For premixtures, the extract is evaporated and the residue is dissolved in DMF/water. The content of diclazuril is determined by ternary gradient reversed-phase high- performance liquid chromatography (HPLC) using a UV detector. 3. Reagents 3. 1. Water, HPLC-grade. 3. 2. Ammonium acetate. 3. 3. Tetrabutylammonium hydrogen sulphate (TBHS). 3. 4. Acetonitrile, HPLC grade. 3. 5. Methanol, HPLC grade. 3. 6. N,N-dimethylformamide (DMF). 3. 7. 3. 8. Hydrochloric acid, q20 = 1,19 g/ml. Standard substance: diclazuril II-24: 1,2,4-triazin-2-yl) phenyl] acetonitrile with guaranteed purity, E771. (+)-4-chlorphenyl [2,6-dichloro-4-(2,3,4,5-tetrahydro-3,5-dioxo- 3. 8. 1. Diclazuril stock standard solution, 500 lg/ml. Weigh to the nearest 0,1 mg, 25 mg of diclazuril standard substance (3. 8) in a 50 ml graduated flask. Dissolve in DMF (3. 6), make up to the mark with DMF (3. 6) and mix. Wrap the flask with aluminium foil or use amber flask and store in the refrigerator. At a temperature of @ 4 (cid:176)C the solution is stable for one month. 6. 5. 1999 EN Official Journal of the European Communities L 118/43 3. 8. 2. Diclazuril standard solution, 50 lg/ml. Transfer 5,00 ml of the stock standard solution (3. 8. 1. ) into a 50 ml graduated flask, make up to the mark with DMF (3. 6) and mix. Wrap the flask with aluminium foil or use amber flask and store in the refrigerator. At a temperature of @ 4 (cid:176)C the solution is stable for one month. 3. 9. Internal standard substance: 2,6 dichloro-a-(4-chlorophenyl)-4-(4,5 dihydro-3,5-dioxo-1,2,4-triazine-2 (3H) - yl) a-methylbenzene-acetonitrile. 3. 9. 1. Internal stock standard solution, 500 lg/ml. Weigh to the nearest 0,1 mg 25 mg of internal standard substance (3. 9) in a 50 ml graduated flask. Dissolve in DMF (3. 6), make up to the mark with DMF (3. 6) and mix. Wrap the flask with aluminium foil or use amber flask and store in the refrigerator. At a temperature of @ 4 (cid:176)C the solution is stable for one month. 3. 9. 2. Internal standard solution, 50 lg/ml. Transfer 5,00 ml of the internal stock standard solution (3. 9. 1) into a 50 ml graduated flask, make up to the mark with DMF (3. 6) and mix. Wrap the flask with aluminium foil or use amber flask and store in the refrigerator. At a temperature of @ 4 (cid:176)C the solution is stable for one month. 3. 9. 3. Internal standard solution for premixtures, p/1000 mg/ml (p = nominal content of diclazuril in the premixture in mg/kg). Weigh to the nearest 0,1 mg p/10 mg of the internal standard substance in a 100 ml graduated flask, dissolve in DMF (3. 6) in a ultrasonic bath (4. 6), make up to the mark with DMF and mix. Wrap the flask with aluminium foil or use amber flask and store in a refrigerator. At a temperature of @ 4 (cid:176)C the solution is stable for one month. 3. 10. Calibration solution, 2 lg/ml. Pipet 2,00 ml diclazuril standard solution (3. 8. 2) and 2,00 ml internal standard solution (3. 9. 2) into a 50 ml graduated flask. Add 16 ml DMF (3. 6), make up to the mark with water and mix. This solution must be prepared freshly before use. 3. 11. C18 solid phase extraction cartridge, e. g. Bond Elut, size: 1 cc, sorbent mass: 100 mg. 3. 12. Extraction solvent: acidified methanol. Pipet 5,0 ml hydrochloric acid (3. 7) into 1000 ml of methanol (3. 5), and mix. 3. 13. Mobile phase for HPLC. Eluent A: ammonium acetate - tetrabutylammonium hydrogen sulphate solution. 3. 13. 1. Dissolve 5 g ammonium acetate (3. 2) and 3,4 g TBHS (3. 3) in 1000 ml water (3. 1) and mix. 3. 13. 2. Eluent B: acetonitrile (3. 4). 3. 13. 3. Eluent C: methanol (3. 5). 4. Apparatus 4. 1. Mechanical shaker. 4. 2. Equipment for ternary gradient HPLC. 4. 2. 1. Liquid chromatographic column, Hypersil ODS, 3 lm packing, 100 mm x 4,6 mm, or equivalent. 4. 2. 2. UV detector with variable wavelength adjustment or diode array detector. 4. 3. Vacuum rotary evaporator 4. 4. Membrane filter, 0,45 lm. L 118/44 EN Official Journal of the European Communities 6. 5. 1999 4. 5. Vacuum manifold. 4. 6. Ultrasonic bath. 5. Procedure 5. 1. General 5. 1. 1. Blank feed A blank feed should be analysed to check that neither diclazuril nor interfering substances are present. The blank feed should be similar in type to that of the sample and on analysis diclazuril or interfering substances should not be detected. 5. 1. 2. Recovery test A recovery test should be carried out by analysing the blank feed which has been fortified by addition of a quantity of diclazuril similar to that present in the sample. To fortify at a level of l mg/kg add 0,1 ml of the stock standard solution (3. 8. 1. ) to 50 g of a blank feed, mix thoroughly and leave for 10 min, mixing again several times before proceeding (5. 2. ). Alternatively, if a blank feed similar in type to that of the sample is not available (see 5. 1. 1), a recovery test can be performed by means of the standard addition method. In this case, the sample to be analysed is fortified with a quantity of diclazuril, similar to that already present in the sample. This sample is analysed, together with the unfortified sample and the recovery can be calculated by subtraction. 5. 2. Extraction 5. 2. 1. Feedingstuffs Weigh to the nearest 0,01 g approximately 50 g of the sample. Transfer to a 500 ml conical flask, add 1,00 ml internal standard solution (3. 9. 2), 200 ml extraction solvent (3. 12) and stopper the flask. Shake the mixture on the shaker (4. 1) overnight. Allow to settle for 10 minutes. Transfer a 20 ml aliquot of the supernatant to a suitable glass container and dilute with 20 ml water. Transfer this solution on an extraction cartridge (3. 11), and pass through by applying vacuum (4. 5. ). Wash the cartridge with 25 ml of a mixture of extraction solvent (3. 12) and water, 65 + 35 (V + V). Discard the collected fractions and elute the compounds with 25 ml of a mixture of extraction solvent (3. 12) and water, 80 + 20 (V + V). Evaporate this fraction until it had just reached dryness by means of the rotary evaporator (4. 3) at 60 (cid:176)C. Dissolve the residue in 1,0 ml DMF (3. 6), add 1,5 ml of water (3. 1) and mix. Filter through a membrane filter (4. 4). Proceed to the HPLC determination (5. 3). 5. 2. 2. Premixtures Weigh to the nearest 0,001 g approximately 1 g of the sample. Transfer to a 500 ml conical flask, add 1,00 ml internal standard solution (3. 9. 3), 200 ml extraction solvent (3. 12) and stopper the flask. Shake the mixture overnight on the shaker (4. 1). Allow to settle for 10 minutes. Transfer an aliquot of 10. 000/p ml (p = nominal content of diclazuril in the premix in mg/kg) of the supernatant to a round bottomed flask of suitable size. Evaporate until it had just reached dryness, under reduced pressure at 60 (cid:176)C by means of the rotary evaporator (4. 3). Redissolve the residue in 10,0 ml DMF (3. 6), add 15,0 ml water (3. 1) and mix. Proceed to the HPLC determination (5. 3). 5. 3. HPLC determination 5. 3. 1. Parameters The following conditions are offered for guidance, other conditions may be used provided that they give equivalent results. (cid:190) Liquid chromatographic 100 mm · 4,6 mm, Hypersil ODS, 3 lm packing, or equivalent column (4. 2. 1. ): (cid:190) Mobile phase: Eluent A (3. 13. 1): Aqueous acetate hydrogen sulphate and solution of ammonium tetrabutyl-ammonium Eluent B (3. 13. 2): acetonitrile Eluent C (3. 13. 3): methanol 6. 5. 1999 EN Official Journal of the European Communities L 118/45 (cid:190) Elution mode: (cid:190) linear gradient (cid:190) initial conditions: A+B+C = 60+20+20 (v+v+v) (cid:190) after 10 minutes gradient elution for 30 minutes to: A+B+C = 45+20+35 (v+v+v) Flush with B for 10 minutes (cid:190) Flow rate: 1,5 – 2 ml/min (cid:190) Injection volume: (cid:190) Detector wavelength: 20 ll 280 ml Check the stability of the chromatographic system, injecting several times the calibration solution (3. 10), containing 2 lg/ml, until constant peak heights and retention times are achieved. 5. 3. 2. Calibration solution Inject 20 ll of the calibration solution (3. 10) several times and determine the mean peak height (area) of the diclazuril and internal standard peaks. 5. 3. 3. Sample solution Inject 20 ll of the sample solution (5. 2. 1. or 5. 2. 2. ) several times and determine the mean peak height (area) of the diclazuril and internal standard peaks. 6. Calculation of the results 6. 1. Feeds The diclazuril content w (mg/kg) in the sample is given by the following formula: w = hd,s. hi,c hi,s. hd,c. bd,c. 10V m [mg/kg] where h d,s = peak height (area) of diclazuril in the sample solution (5. 2. 1) h = peak height (area) of the internal standard in the sample solution (5. 2. 1) i,s h d,c h i,c = peak height (area) of diclazuril in the calibration solution (3. 10) = peak height (area) of the internal standard in the calibration solution (3. 10) b d,c = diclazuril concentration in the calibration solution in lg/ml (3. 10) m = mass of the test portion in g. V = volume of the sample extract according to 5. 2. 1 (i. e. 2,5 ml) 6. 2. Premixtures The diclazuril content w (mg/kg) in the sample is given by the formula: w = hd,s. hi,c. h h d,c i,s. bd,c. 0,02V. p m [mg/kg] where hd,c = peak height (area) of diclazuril in the calibration solution (3. 10) hi,c = peak height (area) of the internal standard in the calibration solution (3. 10) hd,s = peak height (area) of diclazuril in the sample solution (5. 2. 2) hi,s = peak height (area) of the internal standard in the sample solution (5. 2. 2) bd,c = diclazuril concentration in the calibration solution (3. 10) m = mass of the test portion in g. V = volume of the sample extract according to 5. 2. 2 (i. e. 25 ml) p = nominal content of diclazuril in mg/kg in the premixture L 118/46 EN Official Journal of the European Communities 6. 5. 1999 7. Validation of the results 7. 1. Identity The identity of the analyte can be confirmed by co-chromatography, or by using a diode-array detector by which the spectra of the sample extract (5. 2. 1 or 5. 2. 2) and the calibration solution (3. 10) are compared. 7. 1. 1. Co-chromatography A sample extract (5. 2. 1 or 5. 2. 2) is fortified by addition of an appropriate amount of calibration solution (3. 10). The amount of added diclazuril should be similar to the amount of diclazuril found in the sample extract. Only the height of the diclazuril peak and the internal standard peak should be enhanced after taking into account both the amount added and the dilution of the extract. The peak width, at half of its height, must be within 5 10 % of the original width of the diclazuril peak or the internal standard peak of the unfortified sample extract. 7. 1. 2. Diode-array detection The results are evaluated according to the following criteria: (a) The wavelength of maximum absorption of the sample and of the standard spectra, recorded at the peak apex on the chromatogram, must be the same within a margin determined by the resolving power of the detection system. For diode-array detection this is typically within 5 2 nm. (b) Between 230 and 320 nm, the sample and standard spectra recorded at the peak apex of the chromatogram, must not be different for those parts of the spectrum within the range 10-100 % of relative absorbance. This criterion is met when the same maxima are present and at no observed point the deviation between the two spectra exceeds 15 % of the absorbance of the standard analyte. (c) Between 230 and 320 nm, the spectra of the upslope, apex and downslope of the peak produced by the sample extract must not be different from each other for those parts of the spectrum within the range 10-100 % of relative absorbance. This criterion is met when the same maxima are present and when at all observed points the deviation between the spectra does not exceed 15 % of the absorbance of the spectrum of the peak apex. If one of these criteria is not met the presence of the analyte has not been confirmed. 7. 2. Repeatability The difference between the results of two parallel determinations carried out on the same sample must not exceed: (cid:190) 30 % relative, to the higher value for diclazuril contents from 0,5 mg/kg to 2,5 mg/kg (cid:190) 0,75 mg/kg for diclazuril contents between 2,5 mg/kg and 5 mg/kg (cid:190) 15 % relative to the higher value for diclazuril contents of more than 5 mg/kg 7. 3. Recovery For a fortified (blank) sample the recovery should be at least 80 %. 8. Results of a collaborative study A collaborative study was arranged in which five samples were analysed by 11 laboratories. These samples consisted of two premixtures; one was mixed with an organic matrix (O 100) and the other with an inorganic matrix (A 100). The theoretical content is 100 mg diclazuril per kg. The three mixed feeds for poultry were made by three different producers (NL) (L1/Z1/K1). The theoretical content is 1 mg diclazuril per kg. The laboratories were instructed to analyse each of the samples once or in duplicate. (More detailed information on this collaborative study can be found in the Journal of AOAC International, Volume 77, No 6, 1994, p. 1359-1361). The results are given in the following table: 6. 5. 1999 EN Official Journal of the European Communities L 118/47 Sample 1 A 100 Sample 2 O 100 Sample 3 L 1 Sample Z 1 Sample 5 K 1 11 19 0,89 0,15 17,32 0,17 18,61 1 11 19 1,15 0,02 1,92 0,11 9,67 1 6 12 0,89 0,03 3,34 0,12 13,65 1 L n Mean S (mg/kg) (%) r (mg/kg) (%) R r CV S R CV 11 19 100,8 5,88 5,83 7,59 7,53 11 18 103,5 7,64 7,38 7,64 7,38 Nominal content (mg/kg) 100 100 L = number of laboratories. n = number of single values. sr = Standard deviation of repeatability. CV SR = standard deviation of reproducibility. CVR = coefficient of variation of reproductibility. r = coefficient of variation of repeatability. 9. Observations The diclazuril response must have been previously demonstrated to be linear over the range of concentrations being measured. Part C DETERMINATION OF CARBADOX Methyl 3-(2-quinoxalinylmethylene)carbazate N1,N4-dioxide 1. Purpose and scope This method is for the determination of carbadox in feedingstuffs, premixtures and preparations. The detection limit is 1 mg/kg. The limit of determination is 10 mg/kg. 2. Principle The sample is equilibrated with water and extracted with methanol-acetonitrile. For feedingstuffs, an aliquot portion of the filtered extract is subjected to clean-up on an aluminium oxide column. For premixtures and preparations an aliquot portion of the filtered extract is diluted to an appropriate concentration with water, methanol and acetonitrile. The content of carbadox is determined by reversed-phase highperformance liquid chromatography (HPLC) using a UV detector. 3. 3. 1. 3. 2. 3. 3. 3. 4. 3. 5. Reagents Methanol. Acetonitrile, HPLC grade. Acetic acid, w = 100 %. Aluminium oxide: neutral, activity grade I. Methanol-acetonitrile 1+l (v+v). Mix 500 ml of methanol (3. 1) with 500 ml of acetonitrile (3. 2). 3. 6. Acetic acid, s = 10 %. Dilute 10 ml acetic acid (3. 3) to 100 ml with water. 3. 7. Sodium acetate, CH 3 COONa. L 118/48 EN Official Journal of the European Communities 6. 5. 1999 3. 8. Water, HPLC grade. 3. 9. Acetate buffer solution, c = 0,01 mol/l, pH = 6,0. Dissolve 0,82 g of sodium acetate (3. 7) in 700 ml of water (3. 8) and adjust the pH to 6,0 with acetic acid (3. 6). Transfer to a 1 000 ml graduated flask, make up to the mark with water (3. 8) and mix. 3. 10. Mobile phase for HPLC. Mix 825 ml of acetate buffer solution (3. 9) with 175 ml of acetonitrile (3. 2). Filter through a 0,22 lm filter (4. 5) and degas the solution (e. g. by ultrasonification for 10 minutes). 3. 11. Standard substance. Pure carbadox: Methyl 3-(2-quinoxalinylmethylene)carbazate N1,N4-dioxide, E 850. 3. 11. 1. Carbadox stock standard solution, 100 lg/ml (see Point 5 Procedure): Weigh to the nearest 0,1 mg, 25 mg of carbadox standard substance (3. 11) into a 250 ml graduated flask. Dissolve in methanol-acetonitrile (3. 5) by ultrasonification (4. 7). After ultrasonic treatment bring the solution to room temperature, make up to the mark with methanol-acetonitrile (3. 5) and mix. Wrap the flask with aluminium foil or use amber glassware and store in a refrigerator. At a temperature of @ 4 (cid:176)C the solution is stable for one month. 3. 11. 2. Calibration solutions Transfer 2,0, 5,0, 10,0, and 20,0 ml of the stock standard solution (3. 11. 1) into a series of 100 ml calibrated flasks. Add 30 ml of water, make up to the mark with methanol-acetonitrile (3. 5) and mix. Wrap the flask with aluminium foil. These solutions correspond to 2,0, 5,0, 10,0 and 20,0 lg/ml of carbadox respectively. Calibration solutions must be freshly prepared before use. Note: For the determination of carbadox in feedingstuffs containing less than 10 mg/kg, calibration solutions with a concentration below 2,0 lg/ml must be prepared. 3. 12. Water-[methanol-acetonitrile] (3. 5) mixture, 300+700 (v+v) Mix 300 ml of water with 700 ml of the mixture of methanol-acetonitrile (3. 5). 4. Apparatus 4. 1. Laboratory shaker or magnetic stirrer. 4. 2. Glass fibre filter paper (Whatman GF/A or equivalent). 4. 3. Glass column (length 300 to 400 mm, internal diameter approximately 10 mm) with sintered glass frit and draw-off valve. Note: a glass column fitted with a stopcock or a glass column with a tapered end may also be used; in this case, a small glass-wool plug is inserted into the lower end and it is tamped down using a glass rod 4. 4. HPLC equipment with injection system, suitable for injection volumes of 20 ll. 4. 4. 1. Liquid chromatographic column: 300 mm x 4 mm, C18, 10 lm packing or equivalent. 4. 4. 2. UV detector with variable wavelength adjustment or diode array detector operating in the range of 225 to 400 nm. 4. 5. Membrane filter, 0. 22 lm. 4. 6. Membrane filter, 0. 45 lm. 4. 7. Ultrasonic bath. 5. Procedure Note: Carbadox is light-sensitive. Carry out all procedures under subdued light or use amber glassware or glassware wrapped in aluminium foil. 5. 1. General 6. 5. 1999 EN Official Journal of the European Communities L 118/49 5. 1. 1. Blank feed. For the performance of the recovery test (5. 1. 2) a blank feed should be analysed to check that neither carbadox nor interfering substances are present. The blank feed should be similar in type to that of the sample and on analysis carbadox or interfering substances should not be detected. 5. 1. 2. Recovery test. A recovery test should be carried out by analysing the blank feed (5. 1. 1) which has been fortified by the addition of a quantity of carbadox, similar to that present in the sample. To fortify at a level of 50 mg/kg, transfer 5,0 ml of the stock standard solution (3. 11. 1) to a 200 ml conical flask. Evaporate the solution to approximately 0,5 ml in a stream of nitrogen. Add 10 g of the blank feed, mix and wait for 10 minutes before proceeding with the extraction step (5. 2). Alternatively, if a blank feed similar in type to that of the sample is not available (see 5. 1. 1), a recovery test can be performed by means of the standard addition method. In this case, the sample is fortified with a quantity of carbadox, similar to that already present in the sample. This sample is analysed, together with the unfortified sample and the recovery can be calculated by subtraction. 5. 2. Extraction 5. 2. 1. Feedingstuffs. Weigh to the nearest 0,01 g, approximately 10 g of the sample and transfer to a 200 ml conical flask. Add 15,0 ml of water, mix, and equilibrate for 5 minutes. Add 35,0 ml of methanol-acetonitrile (3. 5), stopper and shake for 30 minutes on the shaker or stir on the magnetic stirrer (4. 1). Filter the solution through a glass fibre filter paper (4. 2). Retain this solution for the purification step (5. 3). 5. 2. 2. Premixtures (0,1 to 2,0 %). Weigh to the nearest 0,01 g, approximately l g of the unground sample and transfer to a 200 ml conical flask. Add l5, 0 ml of water, mix, and equilibrate for 5 minutes. Add 35,0 ml of methanol-acet- onitrile (3. 5), stopper and shake for 30 minutes on the shaker or stir on the magnetic stirrer (4. 1). Filter the solution through a glass fibre filter paper (4. 2). Pipet an aliquot of filtrate into a 50 ml calibrated flask. Add 15,0 ml of water, make up to the mark with methanol-acetonitrile (3. 5) and mix. The carbadox concentration of the final solution should be approximately 10 lg/ml. An aliquot is filtered through a 0,45 lm filter (4. 6). Proceed to the HPLC determination (5. 4). 5. 2. 3. Preparations (; 2 %) Weigh to the nearest 0,001 g, approximately 0,2 g of the unground sample and transfer to a 250 ml conical flask. Add 45,0 ml of water, mix, and equilibrate for 5 minutes. Add 105,0 ml of methanol- acetonitrile (3. 5), stopper and homogenise. Sonicate (4. 7) the sample for 15 minutes followed by shaking or stirring for 15 minutes (4. 1). Filter the solution through a glass fibre filter paper (4. 2). Dilute an aliquot of filtrate with the mixture of water-methanol-acetonitrile (3. 12) to a final carbadox concentration of 10-15 lg/ml (for a 10 % preparation, the dilution factor is 10). An aliquot is filtered through a 0,45 lm filter (4. 6). Proceed to the HPLC determination (5. 4). 5. 3. Purification 5. 3. 1. Preparation of the aluminium oxide column. Weigh 4 g of aluminium oxide (3. 4) and transfer it to the glass column (4. 3). 5. 3. 2. Sample purification. Apply 15 ml of the filtered extract (5. 2. 1) to the aluminium oxide column and discard the first 2 ml of eluate. Collect the next 5 ml and filter an aliquot through a 0. 45 lm filter (4. 6). Proceed to the HPLC determination (5. 4). 5. 4. HPLC determination L 118/50 EN Official Journal of the European Communities 6. 5. 1999 5. 4. 1. Parameters The following conditions are offered for guidance, other conditions may be used provided they yield equivalent results. Liquid chromatographic column (4. 1. 1): 300 mm · 4 mm, C18, 10 lm packing or equivalent. Mobile phase (3. 10): Flow rate: Detection wavelength: Injection volume: Mixture of acetate buffer solution (3. 9) and acetonitrile (3. 2), 825+175 (v+v). 1,5-2 ml/min. 365 nm. 20 ll. injecting the calibration solution (3. 11. 2) Check the stability of containing 5,0 lg/ml several times, until constant peak heights (areas) and retention times are achieved. the chromatographic system, 5. 4. 2. Calibration graph. Inject each calibration solution (3. 11. 2) several times and measure the peak heights (areas) for each concentration. Plot a calibration curve using the mean peak heights or areas of the calibration solutions as the ordinates and corresponding concentrations in lg/ml as the abscissae. 5. 4. 3. Sample solution. Inject the sample extract [(5. 3. 2) for feedingstuffs, (5. 2. 2) for premixtures and (5. 2. 3) for preparations] several times and determine the mean peak height (area) of the carbadox peaks. 6. Calculation of the results From the mean height (area) of the carbadox peaks of the sample solution determine the concentration of the sample solution in lg/ml by reference to the calibration graph (5. 4. 2). 6. 1. Feedingstuffs: The content of carbadox w (mg/kg) in the sample is given by the following formula: w = b · V1 m [mg/kg] in which: b = carbadox concentration of the sample extract (5. 3. 2) in lg/ml. V1 = extraction volume in ml (i. e. 50). m = mass of the test portion in g. 6. 2. Premixutres and preparations. The content of carbadox w (mg/kg) in the sample is given by the following formula: w = · f b · V 2 m [mg/kg] in which: b = carbadox concentration of the sample extract (5. 2. 2 or 5. 2. 3) in lg/ml. V2 = extraction volume in ml (i. e. 50 for premixtures; 150 for preparations). f = dilution factor according to 5. 2. 2 (premixtures) or 5. 2. 3 (preparations). m = mass of the test portion in g. 6. 5. 1999 EN Official Journal of the European Communities L 118/51 7. Validation of the results 7. 1. Identity. The identity of the analyte can be confirmed by co-chromatography, or by using a diode-array detector by which the spectra of the sample extract and the calibration solution (3. 11. 2) containing 10,0 lg/ml are compared. 7. 1. 1. Co-chromatography. A sample extract is fortified by addition of an appropriate amount of calibration solution (3. 11. 2). The amount of added carbadox should be similar to the estimated amount of carbadox found in the sample extract. Only the height of the carbadox peak should be enhanced after taking into account both the amount added and the dilution of the extract. The peak width, at half of its maximum height, must be within approximately 10 % of the original width. 7. 1. 2. Diode-array detection. The results are evaluated according to the following criteria: (a) the wavelength of maximum absorption of the sample and of the standard spectra, recorded at the peak apex on the chromatogram, must be the same within a margin determined by the resolving power of the detection system. For diode-array detection, this is typically within 5 2 nm; (b) between 225 and 400 nm, the sample and standard spectra recorded at the peak apex on the chromatogram, must not be different for those parts of the spectrum within the range 10 to 100 % of relative absorbance. This criterion is met when the same maxima are present and at no observed point the deviation between the two spectra exceeds 15 % of the absorbance of the standard analyte; (c) between 225 and 400 nm, the spectra of the upslope, apex and downslope of the peak produced by the sample extract must not be different from each other for those parts of the spectrum within the range 10 to 100 % of relative absorbance. This criterion is met when the same maxima are present and when at all observed points the deviation between the spectra does not exceed 15 % of the absorbance of the spectrum of the apex. If one of these criteria is not met the presence of the analyte has not been confirmed. 7. 2. Repeatability. For contents of 10 mg/kg and higher, the difference between the results of two parallel determinations carried out on the same sample must not exceed 15 % relative to the higher result. 7. 3. Recovery. For a fortified (blank) sample the recovery should be at least 90 %. 8. Results of a collaborative study A collaborative study was arranged in which six feedingstuffs, four premixtures and three preparations were analysed by eight laboratories. Duplicate analyses were performed on each sample. (More detailed information on this collaborative study can be found in the Journal of the AOAC, Volume 71, 1988, p. 484-490). The results (excluding outliers) are shown below: Table 1. Results of the collaborative study for feedingstuffs L n Mean (mg/kg) (mg/kg) S r CVr (%) S (mg/kg) R CVR (%) Nominal content (mg/kg) Sample 1 Sample 2 Sample 3 Sample 4 Sample 5 Sample 6 8 15 50,0 2,90 5,8 3,92 7,8 8 14 47,6 2,69 5,6 4,13 8,7 8 15 48,2 1,38 2,9 2,23 4,6 8 15 49,7 1,55 3,1 2,58 5,2 8 15 46,9 1,52 3,2 2,26 4,8 8 15 49,7 2,12 4,3 2,44 4,9 50,0 50,0 50,0 50,0 50,0 50,0 L 118/52 EN Official Journal of the European Communities 6. 5. 1999 Table 2. Results of the collaborative study for premixtures and preparations Premixtures Preparations A B C D 7 14 8,89 0,37 4,2 0,37 4,2 7 14 9,29 0,28 3,0 0,28 3,0 7 14 9,21 0,28 3,0 0,40 4,3 7 14 8,76 0,44 5,0 0,55 6,3 A 8 16 94,6 4,1 4,3 5,4 5,7 B 8 16 98,1 5,1 5,2 6,4 6,5 C 8 16 104 7,7 7,4 7,7 7,4 L n Mean (g/kg) (g/kg) S r CV (%) r SR (g/kg) (%) CV R Nominal content (g/kg) 10,0 10,0 10,0 10,0 100 100 100 L: number of laboratories. n: number of single values. s standard deviation of repeatability. : r CVr: coefficient of variation of repeatability. standard deviation of reproductibility. : SR CV : coefficient of variation of reproducibility. R


title: EC economic data pocket book. No 2 /1999.

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Japan
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exchange rate
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title: Directiva 1999/26/CE a Comisiei din 20 aprilie 1999 de adaptare la progresul tehnic a Directivei 93/94/CEE a Consiliului privind spațiul destinat montării plăcuței de înmatriculare din spate la autovehiculele cu două sau trei roți (Text cu relevanță pentru SEE)

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text: 12 RO Jurnalul Oficial al Uniunii Europene 07/vol. 7 31999L0026 6. 5. 1999 JURNALUL OFICIAL AL COMUNITĂȚILOR EUROPENE L 118/32 DIRECTIVA 1999/26/CE A COMISIEI din 20 aprilie 1999 de adaptare la progresul tehnic a Directivei 93/94/CEE a Consiliului privind spațiul destinat montării plăcuței de înmatriculare din spate la autovehiculele cu două sau trei roți (Text cu relevanță pentru SEE) COMISIA COMUNITĂȚILOR EUROPENE, ADOPTĂ PREZENTA DIRECTIVĂ: având în vedere Tratatul de instituire a Comunității Europene, Articolul 1 având în vedere Directiva 92/61/CEE a Consiliului din 30 iunie 1992 privind omologarea autovehiculelor cu două sau trei roți (1), modificată de Actul de aderare a Austriei, Finlandei ș(cid:13)i Suediei, în special articolul 16, având în vedere Directiva 93/94/CEE a Consiliului din 29 octom- brie 1993 privind spațiul destinat montării plăcuței de înmatricu- lare din spate la autovehiculele cu două sau trei roți (2), în special articolul 3, (1) (2) (3) (4) întrucât Directiva 93/94/CEE este una dintre directivele dis- tincte ale procedurii comunitare de omologare introduse prin Directiva 92/61/CEE; întrucât dispozițiile Directivei 92/61/CEE privind sistemele, componente ș(cid:13)i unitățile teh- nice ale vehiculelor se aplică în consecință ș(cid:13)i acelei directive; întrucât dezvoltarea tehnologică permite acum o adaptare a Directivei 93/94/CEE a Consiliului la progresul tehnic; întrucât, pentru a asigura funcționarea corectă a sistemu- lui de omologare completă, este necesar să se clarifice sau să se completeze anumite dispoziții ale respectivei directive; întrucât, în acest sens, este necesară adaptarea dispozițiilor referitoare la condițiile de încărcare ale vehiculelor atunci când se măsoară înclinația ș(cid:13)i cele referitoare la dimensiunile spațiilor destinate montării plăcuței de înmatriculare din spate la cvadricicluri prevăzute cu o caroserie, precum ș(cid:13)i pentru alinierea figurii 1 la poziția reală a vehiculelor în timpul testelor ș(cid:13)i pentru o mai bună menționare a anumi- tor referințe în formularul descriptiv; întrucât măsurile stabilite în această directivă sunt în conformitate cu avizul Comitetului pentru adaptare la progresul tehnic înființat prin articolul 13 din Directiva 70/156/CEE a Consiliului (3), modificată ultima dată de Directiva 98/91/CE a Parlamentului European ș(cid:13)i a Consiliului (4), (1) (2) (3) (4) JO L 225, 10. 8. 1992, p. 72. JO L 311, 14. 12. 1993, p. 83. JO L 42, 23. 2. 1970, p. 1. JO L 11, 16. 1. 1999, p. 25. Anexa la Directiva 93/94/CEE se modifică prin prezenta în conformitate cu anexa la prezenta directivă. Articolul 2 (1) În ceea ce priveș(cid:13)te spațiul destinat montării plăcuței de înma- triculare din spate, începând cu 1 ianuarie 2000, statele membre nu pot: — refuza să acorde omologarea CE pentru un tip de autovehicul cu două sau trei roți; — interzice înmatricularea, vânzarea sau punerea în circulație a autovehiculelor cu două sau trei roți, dacă spațiul destinat montării plăcuței de înmatriculare din spate respectă cerințele Directivei 93/94/CEE, modificată de prezenta directivă. (2) Cu începere de la 1 ianuarie 2000, statele membre refuză să acorde omologarea CE pentru orice fel de autovehicul cu două sau trei roți dacă nu sunt respectate cerințele Directivei 93/94/CEE, modificată de prezenta directivă, privind spațiul destinat montării plăcuței de înmatriculare din spate. Articolul 3 (1) Statele membre adoptă ș(cid:13)i publică, până cel târziu la data de 31 decembrie 1999, dispozițiile necesare aducerii la îndeplinire a prezentei directive. Acestea informează imediat Comisia cu pri- vire la acestea. Statele membre aplică respectivele dispoziții de la 1 ianuarie 2000. La adoptarea de către statele membre a acestor dispoziții, acestea includ o trimitere la prezenta directivă sau sunt însoțite de o ase- menea trimitere în momentul publicării lor oficiale. Statele membre stabilesc modul în care urmează să fie făcută această trimitere. (2) Comisiei îi sunt comunicate de către statele membre dispozițiile de drept intern pe care le adoptă în domeniul regle- mentat de prezenta directivă. 07/vol. 7 RO Jurnalul Oficial al Uniunii Europene 13 Prezenta directivă intră în vigoare în a douăzecea zi de la publicarea în Jurnalul Oficial al Comunităților Europene. Articolul 4 Prezenta directivă se adresează statelor membre. Articolul 5 Adoptată la Bruxelles, 20 aprilie 1999. Pentru Comisie Martin BANGEMANN Membru al Comisiei 14 RO Jurnalul Oficial al Uniunii Europene 07/vol. 7 ANEXA 1. Punctul 1. 1 se înlocuieș(cid:13)te cu următorul text: „1. 1. Mopede ș(cid:13)i cvadricicluri uș(cid:13)oare fără caroserie. ” 2. Punctul 1. 2 se înlocuieș(cid:13)te cu următorul text: „1. 2. Motociclete, tricicluri cu o putere maximă de 15 kW ș(cid:13)i cvadricicluri, altele decât cele uș(cid:13)oare, fără caroserie. ” 3. Punctul 1. 3 se înlocuieș(cid:13)te cu următorul text: „1. 3. Tricicluri cu o putere maximă mai mare de 15 kW, cvadricicluri uș(cid:13)oare prevăzute cu o caroserie ș(cid:13)i cvadricicluri, altele decât cele uș(cid:13)oare prevăzute cu o caroserie. ” 4. Punctul 3. 1. 2 se înlocuieș(cid:13)te cu următorul text: „3. 1. 2. poate să fie înclinată față de verticală cu nu mai mult de 30o, cu vehiculul neîncărcat, atunci când plăcuța de suport a numărului de înmatriculare este orientată în sus;”. 5. Punctul 3. 1. 3 se înlocuieș(cid:13)te cu următorul text: „3. 1. 3. poate să fie înclinată față de verticală cu nu mai mult de 15o, cu vehiculul neîncărcat, atunci când plăcuța de suport a numărului de înmatriculare este orientată în jos;”. 6. Punctul 4. 1 se înlocuieș(cid:13)te cu următorul text: „4. 1. Nici un punct din cadrul spațiului destinat montării plăcuței de înmatriculare nu poate fi cu mai mult de 1,5 m deasupra solului atunci când vehiculul nu este încărcat. ” 7. Punctul 5. 1 se înlocuieș(cid:13)te cu următorul text: „5. 1. Nici un punct din cadrul spațiului destinat montării plăcuței de înmatriculare nu va fi la o înălțime mai mică de 0,20 m sau de raza roții deasupra solului dacă aceasta este mai mică de 0,20 m, atunci când vehiculul nu este încărcat. ” 8. Figura 1 se înlocuieș(cid:13)te cu următoarea: „ Unghiul de vizibilitate geometrică (diedru cu muchie orizontală)” Figura 1 9. Apendicele 1 se înlocuieș(cid:13)te cu următorul text: „Apendicele 1 Formular descriptiv privind spațiul destinat montării plăcuței de înmatriculare din spate la un tip de autovehicul cu două sau trei roți (se anexează la cererea pentru omologarea de tip pentru componente atunci când se depune separat față de cererea pentru omologarea de tip pentru vehicul) 07/vol. 7 RO Jurnalul Oficial al Uniunii Europene 15 Comanda nr. (atribuit de solicitant): Cererea pentru omologarea de tip pentru componente privind spațiul destinat montării plăcuței de înmatriculare din spate la un tip de autovehicul cu două sau trei roți trebuie să conțină informațiile stabilite în anexa II la Directiva 92/61/CEE a Consiliului partea A secțiunile: — 0. 1; — 0. 2; — 0. 4-0. 6; — 2. 2; — 2. 2. 1; — 9. 6; — 9. 6. 1. ”


title: Verordnung (EG) Nr. 821/1999 der Kommission vom 20. April 1999 zur Festsetzung der Mindestverkaufspreise für Rindfleisch für den Verkauf im Rahmen der Ausschreibung nach der Verordnung (EG) Nr. 705/1999

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beef
fixing of prices
minimum price
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text: 21. 4. 1999 DE Amtsblatt der Europ(cid:132)ischen Gemeinschaften L 104/7 VERORDNUNG (EG) Nr. 821/1999 DER KOMMISSION vom 20. April 1999 zur Festsetzung der Mindestverkaufspreise f(cid:129)r Rindfleisch f(cid:129)r den Verkauf im Rahmen der Ausschreibung nach der Verordnung (EG) Nr. 705/1999 DIE KOMMISSION DER EUROP(cid:142)ISCHEN GEMEINSCHAFTEN (cid:190) gest(cid:129)tzt auf den Vertrag zur Gr(cid:129)ndung der Europ(cid:132)ischen Gemeinschaft, gest(cid:129)tzt auf die Verordnung (EWG) Nr. 805/68 des Rates vom 27. Juni 1968 (cid:129)ber die gemeinsame Marktorgani- sation f(cid:129)r Rindfleisch (1), zuletzt ge(cid:132)ndert durch die Verordnung (EG) Nr. 1633/98 (2), insbesondere auf Artikel 7 Absatz 3, in Erw(cid:132)gung nachstehender Gr(cid:129)nde: Durch die Verordnung (EG) Nr. 705/1999 der Kommis- sion (3) sind bestimmte Mengen Rindfleisch ausge- schrieben worden. Nach Artikel 9 der Verordnung (EWG) Nr. 2173/79 der Kommission (4), zuletzt ge(cid:132)ndert durch die Verordnung (EG) Nr. 2417/95 (5), m(cid:129)ssen die Mindestverkaufspreise f(cid:129)r das ausgeschriebene Fleisch aufgrund der eingegan- genen Angebote festgesetzt werden. Die in dieser Verordnung vorgesehenen Maßnahmen entsprechen der Stellungnahme des Verwaltungsaus- schusses f(cid:129)r Rindfleisch (cid:190) HAT FOLGENDE VERORDNUNG ERLASSEN: Artikel 1 Die Mindestverkaufspreise f(cid:129)r Rindfleisch f(cid:129)r die Ausschreibung gem(cid:132)ß der Verordnung (EG) Nr. 705/ 1999, deren Frist f(cid:129)r die Einreichung der Angebote am 12. April 1999 abgelaufen ist, werden im Anhang zu dieser Verordnung festgesetzt. Artikel 2 Diese Verordnung tritt am 21. April 1999 in Kraft. Diese Verordnung ist in allen ihren Teilen verbindlich und gilt unmittelbar in jedem Mitgliedstaat. Br(cid:129)ssel, den 20. April 1999 F(cid:129)r die Kommission Franz FISCHLER Mitglied der Kommission (1) ABl. L 148 vom 28. 6. 1968, S. 24. (2) ABl. L 210 vom 28. 7. 1998, S. 17. (3) ABl. L 89 vom 1. 4. 1999, S. 36. (4) ABl. L 251 vom 5. 10. 1979, S. 12. (5) ABl. L 248 vom 14. 10. 1995, S. 39. L 104/8 DE Amtsblatt der Europ(cid:132)ischen Gemeinschaften 21. 4. 1999 ANEXO (cid:190) BILAG (cid:190) ANHANG (cid:190) PAQAQTGLA (cid:190) ANNEX (cid:190) ANNEXE (cid:190) ALLEGATO (cid:190) B(cid:246)LAGE (cid:190) ANEXO (cid:190) LIITE (cid:190) BILAGA Estado miembro Medlemsstat Mitgliedstaat Jq(cid:145)tor l(cid:146)kor Member State (cid:144)tat membre Stato membro Lidstaat Estado-membro J(cid:132)senvaltio Medlemsstat Productos (1) Produkter (1) Erzeugnisse (1) Pqo(cid:152)(cid:149)mta (1) Products (1) Produits (1) Prodotti (1) Producten (1) Produtos (1) Tuotteet (1) Produkter (1) Precio m(cid:146)nimo expresado en euros por tonelada Mindstepriser i EUR/ton Mindestpreise ausgedr(cid:129)ckt in EUR/Tonne Ek(cid:145)wister pyk(cid:147)seir ejvqaf(cid:149)lemer se Euq(cid:150) am(cid:145) t(cid:149)mo Minimum prices expressed in EUR per tonne Prix minimaux exprim(cid:130)s en euros par tonne Prezzi minimi espressi in euro per tonnellata Minimumprijzen uitgedrukt in euro per ton Pre(cid:135)o m(cid:146)nimo expresso em euros por tonelada V(cid:132)himm(cid:132)ishinnat euroina tonnia kohden ilmaistuna Minimipriser i euro per ton Carne deshuesada (cid:190) Udbenet kød (cid:190) Fleisch ohne Knochen (cid:190) Jq(cid:146)ata wyq(cid:148)r j(cid:149)jaka (cid:190) Boneless beef (cid:190) Viande d(cid:130)soss(cid:130)e (cid:190) Carni senza osso (cid:190) Vlees zonder been (cid:190) Carne desossada (cid:190) Luuton naudanliha (cid:190) Benfritt k(cid:148)tt IRELAND (cid:190) Intervention flank (INT 18) (cid:190) Intervention forequarter (INT 24) UNITED KINGDOM (cid:190) Intervention forequarter (INT 24) (cid:190) Intervention shoulder (INT 22) (cid:190) Intervention brisket (INT 23) (cid:190) 1 212,14 1 210 (cid:190) 750 (1) V(cid:130)anse los anexos V y VII del Reglamento (CEE) no 2456/93 de la Comisi(cid:155)n (DO L 225 de 4. 9. 1993, p. 4), cuya (cid:156)ltima modificaci(cid:155)n la constituye el Reglamento (CE) no 2812/98 (DO L 349 de 24. 12. 1998, p. 47). (1) Se bilag V og VII til Kommissionens forordning (EØF) nr. 2456/93 (EFT L 225 af 4. 9. 1993, s. 4), senest ændret ved forordning (EF) nr. 2812/98 (EFT L 349 af 24. 12. 1998, s. 47). (1) Vgl. Anh(cid:132)nge V und VII der Verordnung (EWG) Nr. 2456/93 der Kommission (ABl. L 225 vom 4. 9. 1993, S. 4), zuletzt ge(cid:132)ndert durch die Verordnung (EG) Nr. 2812/98 (ABl. L 349 vom 24. 12. 1998, S. 47). (1) Bk(cid:146)pe paqaqt(cid:147)lata V jai VII tou jamomislo(cid:151) (EOJ) aqih. 2456/93 tgr Epitqop(cid:147)r (EE L 225 tgr 4. 9. 1993, s. 4), (cid:149)pyr tqopopoi(cid:147)hgje tekeuta(cid:148)a ap(cid:149) tom jamomisl(cid:149) (EJ) aqih. 2812/98 (EE L 349 tgr 24. 12. 1998, s. 47). (1) See Annexes V and VII to Commission Regulation (EEC) No 2456/93 (OJ L 225, 4. 9. 1993, p. 4), as last amended by Regulation (EC) No 2812/98 (OJ L 349, 24. 12. 1998, p. 47). (1) Voir annexes V et VII du r(cid:138)glement (CEE) no 2456/93 de la Commission (JO L 225 du 4. 9. 1993, p. 4). R(cid:138)glement modifi(cid:130) en dernier lieu par le r(cid:138)glement (CE) no 2812/98 (JO L 349 du 24. 12. 1998, p. 47). (1) Cfr. allegati V e VII del regolamento (CEE) n. 2456/93 della Commissione (GU L 225 del 4. 9. 1993, pag. 4), modificato da ultimo dal regolamento (CE) n. 2812/98 (GU L 349 del 24. 12. 1998, pag. 47). (1) Zie de b¶lagen V en VII van Verordening (EEG) nr. 2456/93 van de Commissie (PB L 225 van 4. 9. 1993, blz. 4), laatstelijk gewijzigd bij Verordening (EG) nr. 2812/98 (PB L 349 van 24. 12. 1998, blz. 47). (1) Ver anexos V e VII do Regulamento (CEE) n. o 2456/93 da Comiss(cid:209)o (JO L 225 de 4. 9. 1993, p. 4). Regulamento com a (cid:156)ltima redac(cid:135)(cid:209)o que lhe foi dada pelo Regulamento (CE) n. o 2812/98 (JO L 349 de 24. 12. 1998, p. 47). (1) Katso komission asetuksen (ETY) N:o 2456/93 (EYVL L 225, 4. 9. 1993, s. 4), sellaisena kuin se on viimeksi muutettuna asetuksella (EY) N:o 2812/98 (EYVL L 349, 24. 12. 1998, s. 47) liitteet V ja VII. (1) Se bilagorna V och VII i f(cid:148)rordning (EEG) nr 2456/93 (EGT L 225, 4. 9. 1993, s. 4), senast (cid:132)ndrad genom f(cid:148)rordning (EG) nr 2812/98 (EGT L 349, 24. 12. 1998, s. 47)


title: WRITTEN QUESTION No. 1043/99 by Ilona GRAENITZ Screening the candidate countries' environmental legislation

eurovoc_concepts:
Community acquis
EU environmental policy
accession negotiations
accession to the European Union
environmental law

text: C 370/158 Europeiska gemenskapernas officiella tidning SV 21. 12. 1999 (1999/C 370/223) SKRIFTLIG FRÅGA E-1043/99 från Ilona Graenitz (PSE) till kommissionen (20 april 1999) Angående: Undersökning av kandidatländernas miljölagstiftning Vilket datum har fastställts som referensdatum för undersökningen av kandidatländernas miljölagstiftning? Kommer detta datum att anpassas efter förhandlingarnas längd? Svar från Hans Van den Broek på kommissionens vägnar (7 maj 1999) Genomgången av gemenskapens regelverk på miljöområdet omfattar alla gemenskapsåtgärder som vidta- gits fram till och med den 1 januari 1999. Även de åtgärder som vidtagits av gemenskapen efter detta datum skall gås igenom i syfte att bedöma kandidatländernas införlivande och genomförande av dessa. Kommissionen kommer därför att diskutera de åtgärder som nyligen antagits med kandidatländerna vid en lämplig tidpunkt i förhandlingarna. (1999/C 370/224) SKRIFTLIG FRÅGA E-1060/99 från Carlos Bru Purón (PSE) till kommissionen (20 april 1999) Angående: Växlingsavgifter Som en försiktighetsåtgärd inför framtiden passar vissa banker i euroområdet i år på att ta ut avgifter på ibland mer än 25 % vid växlingen av europeiska valutor. Dessutom tillämpar de ett regressivt kriterium, vilket gör det förhållandevis dyrare att växla små belopp. Tänker kommissionen uppmärksamma bankerna eller bankorganisationerna i de medlemsstater som deltar i EMU på de negativa effekter som denna strävan att i sista sekunden få ut så mycket som möjligt kommer att få för banksektorn i framtiden? Svar från Mario Monti på kommissionens vägnar (7 maj 1999) I Kommissionen önskar informera parlamentsledamoten om att den redan har kontaktat banker och banksammanslutningar avseende de växlingsavgifter som bankerna tar ut för växling av sedlar i de tidigare nationella valutorna inom euroområdet. januari 1999 sände kommissionen ett brev till de största europeiska bankförbunden där man påpekade att allmänhetens förtroende för införandet av euron och förtroendet för banksystemet som helhet inte får undermineras av att finansiella institut tar ut för höga avgifter för transaktioner mellan de tidigare nationella valutorna inom euroområdet. I ett pressmeddelande av den 5 februari 1999 (1) uppmanade kommissionen bankerna att öka insynen genom att göra sådan information tillgänglig som visar förändringarna i den totala avgiftsnivån (före och efter införandet av euron) för växling av sedlar samt gränsöverskridande checkbetalningar, banköverföringar och kontokort- betalningar mellan de tidigare nationella valutorna inom euroområdet. Nyligen gjordes även, genom kommissionens försorg, en analys tillgänglig för allmänheten av de klagomål som kommit in genom den för ändamålet särskilt skapade e-post- och faxtjänst som tillkännagavs i IP/99/90. Banksammanslutningarna har också blivit ombedda att lämna synpunkter på innehållet i analysen. Parlamentsledamoten hänvisas även till kommissionens svar på de skriftliga frågorna E-3825/98 av Gérard Caudron (2) och P-52/99 av Gianni Tamino (3) och till svaren på de muntliga frågorna O-29/99 av Mark


title: SCHRIFTLICHE ANFRAGE Nr. 1000/99 von Ian WHITE Artikel 9 der Verordnung (EWG) Nr. 95/93 des Rates vom 18. Januar 1993

eurovoc_concepts:
United Kingdom
airline
airport
common transport policy
competence of the Member States
liberalisation of the market
public service
transport regulations

text: 21. 12. 1999 DE Amtsblatt der Europäischen Gemeinschaften C 370/151 Antwort von Herrn van den Broek im Namen der Kommission (7. Mai 1999) Die Kommission teilt der Frau Abgeordneten mit, daß Reaktor 3 des Kernkraftwerks Tschernobyl nach einem geplanten Ausfall von drei Monaten zwecks Überprüfung und Instandhaltung am 6. März 1999 den Betrieb wiederaufnahm. In der 1995 unterzeichneten Gemeinsamen Absichtserklärung zwischen den G7-Ländern, der Kommission und der Ukraine verpflichtete sich letztere, das Kernkraftwerk Tschernobyl bis zum Jahr 2000 stillzulegen. Die Kommission ist der Auffassung, daß seit der Gemeinsamen Absichtserklärung von 1995 Fortschritte erzielt wurden und hofft insbesondere, daß die Ukraine ihre Verpflichtung einhalten wird, das Kernkraft- werk Tschernobyl bis zum Ende des Jahres 2000 stillzulegen. (1999/C 370/212) SCHRIFTLICHE ANFRAGE E-1000/99 von Ian White (PSE) an die Kommission (20. April 1999) Betrifft: Artikel 9 der Verordnung (EWG) 95/93 des Rates vom 18. Januar 1993 Kann die Kommission angeben, welche öffentlichen Flugdienstverpflichtungen gemäß Artikel 9 der Verordnung (EWG) 95/93 des Rates (1) vom 18. Januar 1993 zulässig sind und ob der Südwesten des Vereinigten Königreichs zur Zeit dafür in Frage kommt? Falls nicht, kann die Kommission die Gründe dafür bestätigen, da solche Verpflichtungen für den Erhalt der erforderlichen Zeitnischen unerläßlich sind, die derartige Flughäfen zum Überleben brauchen? (1) ABl. L 14 vom 22. 1. 1993, S. 1. (1999/C 370/213) SCHRIFTLICHE ANFRAGE E-1001/99 von Ian White (PSE) an die Kommission (20. April 1999) Betrifft: Änderung der Verordnung (EWG) 95/93 des Rates vom 18. Januar 1993 Kann die Kommission angeben, wie regionale Flughäfen, die zur Zeit nur mit Schwierigkeiten praktikable Zeitnischen auf Luftverkehrsknotenpunkten/Großflughäfen (beispielsweise Frankfurt) erhalten können, von der versprochenen Änderung der Verordnung (EWG) 95/93 des Rates (1) vom 18. Januar 1993 profitieren werden, und dazu detaillierte Angaben machen? (1) ABl. L 14 vom 22. 1. 1993, S. 1. (1999/C 370/214) SCHRIFTLICHE ANFRAGE E-1002/99 von Ian White (PSE) an die Kommission (20. April 1999) Betrifft: Offener kommerzieller Handel mit Zeitnischen auf Flughäfen Ist sich die Kommission darüber im klaren, daß der eventuelle offene kommerzielle Handel mit Zeitnischen auf Flughäfen durch Fluggesellschaften dazu führen wird, daß kleinere regionale Flugdienste aufgrund rein kommerzieller Zwänge aus Großflughäfen verdrängt werden? Steht dies nicht im Widerspruch zu der Politik der Regionalisierung und des Zugangs zum Luftverkehr auf interregionaler Grundlage?


title: INTERROGAZIONE SCRITTA n. 1003/99 dell'on. Ian WHITE Aeroporti regionali

text: C 370/152 Gazzetta ufficiale delle Comunità europee IT 21. 12. 1999 (1999/C 370/215) INTERROGAZIONE SCRITTA E-1003/99 di Ian White (PSE) alla Commissione (20 aprile 1999) Oggetto: Aeroporti regionali trasporto aereo tra gli aeroporti regionali e le aerostazioni maggiori L’Unione europea si è impegnata a rispettare il principio della regionalizzazione. L’elemento fondamentale dello sviluppo delle regioni all’interno dell’Unione è rappresentato dalla necessità di rendere disponibili (ad esempio, collegamenti di Francoforte). Gli aeroporti regionali incontrano difficoltà nell’ottenere bande orarie praticabili presso dette aerostazioni in quanto non sono previsti incentivi commerciali che stimolino queste ultime ad accogliere aeromobili più piccoli a scapito degli aerei con maggiore capienza di passeggeri e più lucrativi. Ciò è chiaramente contrario alla regionalizzazione e determinerà probabilmente il proseguimento della sotto utilizzazione degli aeroporti regionali e l’ulteriore aumento delle pressioni - trainate dalla domanda - sugli aeroporti più grandi. Considerando tale conflitto di interessi, può la Commissione precisare in che modo intende risolvere la questione? Una soluzione potrebbe essere rappresentata dall’obbligo - limitato - per i grandi aeroporti di fornire bande orarie praticabili agli aeroporti e alle compagnie aeree regionali. Tale procedura è utilizzata nel continente Nordamericano, in cui il maggior aeroporto del mondo (Chicago O’Hare) è obbligato ad assegnare il 20 % delle sue bande orarie secondo le modalità precitate. Risposta comune data dal sig. Kinnock in nome della Commissione alle interrogazioni scritte E-1000/99, E-1001/99, E-1002/99 e E-1003/99 (6 maggio 1999) La Commissione è consapevole del fatto che le linee aeree che operano in arrivo e in partenza dagli aeroporti regionali trovano crescente difficoltà ad assicurarsi adeguate bande orarie presso aeroporti principali congestionati. La Commissione sta preparando una proposta per modificare l’attuale regolamento (CEE) 95/93 del Consiglio del 18 gennaio 1993 relativo a norme comuni per l’assegnazione di bande orarie negli aeroporti della Comunità. In tale contesto, si sta valutando la possibilità di introdurre meccanismi atti a tener conto della situazione specifica delle rotte regionali. L’attuale regolamento prevede la possibilità di riservare bande orarie su rotte di importanza vitale in arrivo e in partenza da aeroporti regionali qualora obblighi di servizio pubblico siano stati imposti ai sensi del regolamento (CEE) 2408/92 del Consiglio del 23 luglio 1992 sull’accesso dei vettori aerei della Comunità alle rotte intracomunitarie (1). Gli Stati membri possono pertanto riservare bande orarie al fine di assicurare l’offerta di tali servizi tra gli aeroporti principali e quelli regionali. La Commissione sta valutando se tali disposizioni debbano essere mantenute in vigore o rafforzate. Per quanto riguarda la situazione nel South West del Regno Unito e in tutte le altre regioni comunitarie, spetta alle autorità dello Stato membro interessato e non alla Commissione prendere le decisioni opportune circa l’imposizione di obblighi di servizio pubblico, in conformità delle rispettive politiche regionali e dei trasporti, purché tali obblighi rispettino, relativamente ad ogni singola rotta, i diversi criteri stabiliti dal regolamento. (1) GU L 240 del 24. 8. 1992. (1999/C 370/216) INTERROGAZIONE SCRITTA E-1005/99 di Ian Hudghton (V) alla Commissione (20 aprile 1999) Oggetto: Ricerche in materia di ormoni di sintesi e di origine vegetale Considerando la controversia riguardo al benessere degli animali che circonda la produzione del farmaco sostitutivo ormonale «Premarin», può la Commissione indicare se l’Unione europea attualmente finanzia ricerche relative alla produzione di ormoni alternativi, di sintesi o di origine vegetale, e se si prevede di aumentare detti finanziamenti?