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Food Additives & Contaminants Publication details, including instructions for authors and subscription information: http://www.tandfonline.com/loi/tfac19
Sulphonamide residues in milk of dairy cows following intravenous injection a
B. Roudaut & J. P. Moretain
a
a
Ministère de l'Agriculture et de la Forêt, Centre National d'Etudes Vétérinaires et Alimentaires, Laboratoire des Médicaments Vétérinaires, Javené, Fougères, 35133, France Published online: 10 Jan 2009.
To cite this article: B. Roudaut & J. P. Moretain (1990) Sulphonamide residues in milk of dairy cows following intravenous injection, Food Additives & Contaminants, 7:4, 527-533, DOI: 10.1080/02652039009373915 To link to this article: http://dx.doi.org/10.1080/02652039009373915
PLEASE SCROLL DOWN FOR ARTICLE Taylor & Francis makes every effort to ensure the accuracy of all the information (the “Content”) contained in the publications on our platform. However, Taylor & Francis, our agents, and our licensors make no representations or warranties whatsoever as to the accuracy, completeness, or suitability for any purpose of the Content. Any opinions and views expressed in this publication are the opinions and views of the authors, and are not the views of or endorsed by Taylor & Francis. The accuracy of the Content should not be relied upon and should be independently verified with primary sources of information. Taylor and Francis shall not be liable for any losses, actions, claims, proceedings, demands, costs, expenses, damages, and other liabilities whatsoever or howsoever caused arising directly or indirectly in connection with, in relation to or arising out of the use of the Content. This article may be used for research, teaching, and private study purposes. Any substantial or systematic reproduction, redistribution, reselling, loan, sublicensing, systematic supply, or distribution in any form to anyone is expressly forbidden. Terms & Conditions of access and use can be found at http:// www.tandfonline.com/page/terms-and-conditions
FOOD ADDITIVES AND CONTAMINANTS, 1990, VOL. 7, NO. 4, 5 2 7 - 5 3 3
Sulphonamide residues in milk of dairy cows following intravenous injection B. ROUDAUT and J. P. MORETAIN Ministère de l'Agriculture et de la Forêt, Centre National d'Etudes Vétérinaires et Alimentaires, Laboratoire des Médicaments Vétérinaires, Javené, 35133 Fougères, France
Downloaded by [Monash University Library] at 13:44 13 September 2013
(Received 26 May 1989; revised 12 January 1990; accepted 19 January 1990) An investigation was conducted into the elimination into milk of four sulphonamides (sulphadimidine, sulphamethoxypyridazine, sulphadimethoxine and sulphadoxine) after intravenous injections of drugs available in France. Sulphonamide determination in milk was performed using a high-performance liquid chromatography (HPLC) method with a detection limit of 0.01 µg/ml. These results were used to determine the withdrawal times (two to six milkings) required for these drugs with a tolerance of 0.1 µg/ml as proposed by the European countries. Keywords: sulphonamide, residues, milk, cows
Introduction
Before antibiotics were used in veterinary medicine, parenteral administration of sulphonamides was practically the only means of treating infectious diseases. Despite the discovery of penicillin and the widespread use of antibiotics, these antimicrobial agents have remained as popular, although the use of sulphonamide declined temporarily (Mol 1975). They are probably still the most commonly used antimicrobial agents throughout the world for the treatment of bovine pneumopathies because of their broad spectrum of activity and their low cost (Brugère et al. 1985). Many studies of their elimination into milk after therapeutic treatment of milking cows were conducted. Several methods for sulphonamide assays in milk were described: microbiological methods (Bogaerts etal. 1981), colorimetric methods based on the Bratton and Marshall reaction (Houston and Umstead 1968, Selzer and Banes 1963, Tishler et al. 1968), gas chromatography (Holtmannspotter and Thier 1982), liquid chromatography (Alawi and Rüssel 1981, Petz 1983, Soback et al. 1983, Nouws et al. 1985). The present study examines the elimination into milk of the main sulphonamides used in dairy cows: sulphadimidine (SDM), sulphamethoxypyridazine (SMP), sulphadimethoxine (SDT) and sulphadoxine (SDX) administered intravenously. The analytical method chosen, highperformance liquid chromatography (HPLC), was sensitive enough (0-01 fig/ml) to obtain sufficiently precise withdrawal times. Materials and methods
Animals The experiments were conducted on a herd of 14 dairy cows (Friesan or 0265-203X/90 $3.00 © 1990 Taylor & Francis Ltd.
528
Β. Roudaut and J. P. Moretain
Normande breeds) at different lactating stages. The animals were kept under usual breeding farm conditions and were milked by machine twice a day. The daily milk production ranged from 10 to 22 litres.
Downloaded by [Monash University Library] at 13:44 13 September 2013
Drugs The four drugs were respectively aqueous solution of sodium sulphadimidine (33g/100ml), sodium sulphamethoxypyridazine (25g/100ml), sodium sulphadimethoxine (25 g/100 ml) and sodium sulphadoxine (20 g/100 ml and 4 g/100 ml trimethoprim). They were administrered at the dosages indicated by the manufacturer after checking sulphonamide concentration. Treatment After the morning milking, drugs were administered to five cows by the intravenous route into the jugular vein using the recommended dose: 100 mg/kg for SDM, 75 mg/kg for SMP, 50 mg/kg for SDT, and 20 mg/kg for SDX. Milking The cows were milked twice a day at 8.00 a.m. and 5.00 p.m. For each cow, samples of 100 ml were taken from the total milk obtained from all quarters collected in individual milk cans after the treatment began. The samples were frozen pending analysis. Analysis Sulphonamides were determined by a liquid chromatography method after extraction of milk and clean-up. Apparatus and Chromatographie conditions. The HPLC system consisted of a SP 8700 pump (Spectra-Physics) coupled with a Spectroflow 773 (Kratos) variable UV detector, which was connected to an integrator (Shimadzu-CR IB). The injector was a Rheodyne 7125 with a 50 μΐ loop. The column was a Novapak C 18, 5 μΐη, 150 X 3-9 mm (Waters). The detection of sulphonamides was performed at 265 nm (SDM, SMP) and 271 nm (SDT, SDX). The mobile phase was acetonitrile, 0-01 M ammonium acetate solution pH 6-6 (10:90, v/v) and the solvent rate was 1 ml/min. Sample preparation. Five millilitres of raw milk, to which was added 20 μ\ HC1 12 Ν for SDT, homogenized by ultrasonication, were extracted with 25 ml ethyl acetate for 10 min (rotary shaker REAX 2, Heidolph). After centrifugation for 5 min at 1500 g, 20 ml of the ethyl acetate extract was evaporated to dryness with a rotary evaporator (Büchi). The dry extract was dissolved in 10 ml HC1 Ν and transferred to a centrifuge tube. The aqueous phase was rinsed with 10 ml of méthylène chloride (or hexane for SDT and SDX) adjusted to pH 5 · 5 with 0 · 9 ml of 10 Ν NaOH and 5 ml of 1 M monobasic potassium phosphate (pH 6*0) then extracted twice with 10 ml of dichloromethane. The organic layer was evaporated to dryness with a rotary evaporator and dissolved in 2 ml of the mobile phase. A 50 μΐ aliquot of the solution was injected for HPLC analysis. The analytical technique used led to detection of concentrations
Food Additives & Contaminants Publication details, including instructions for authors and subscription information: http://www.tandfonline.com/loi/tfac19
Sulphonamide residues in milk of dairy cows following intravenous injection a
B. Roudaut & J. P. Moretain
a
a
Ministère de l'Agriculture et de la Forêt, Centre National d'Etudes Vétérinaires et Alimentaires, Laboratoire des Médicaments Vétérinaires, Javené, Fougères, 35133, France Published online: 10 Jan 2009.
To cite this article: B. Roudaut & J. P. Moretain (1990) Sulphonamide residues in milk of dairy cows following intravenous injection, Food Additives & Contaminants, 7:4, 527-533, DOI: 10.1080/02652039009373915 To link to this article: http://dx.doi.org/10.1080/02652039009373915
PLEASE SCROLL DOWN FOR ARTICLE Taylor & Francis makes every effort to ensure the accuracy of all the information (the “Content”) contained in the publications on our platform. However, Taylor & Francis, our agents, and our licensors make no representations or warranties whatsoever as to the accuracy, completeness, or suitability for any purpose of the Content. Any opinions and views expressed in this publication are the opinions and views of the authors, and are not the views of or endorsed by Taylor & Francis. The accuracy of the Content should not be relied upon and should be independently verified with primary sources of information. Taylor and Francis shall not be liable for any losses, actions, claims, proceedings, demands, costs, expenses, damages, and other liabilities whatsoever or howsoever caused arising directly or indirectly in connection with, in relation to or arising out of the use of the Content. This article may be used for research, teaching, and private study purposes. Any substantial or systematic reproduction, redistribution, reselling, loan, sublicensing, systematic supply, or distribution in any form to anyone is expressly forbidden. Terms & Conditions of access and use can be found at http:// www.tandfonline.com/page/terms-and-conditions
FOOD ADDITIVES AND CONTAMINANTS, 1990, VOL. 7, NO. 4, 5 2 7 - 5 3 3
Sulphonamide residues in milk of dairy cows following intravenous injection B. ROUDAUT and J. P. MORETAIN Ministère de l'Agriculture et de la Forêt, Centre National d'Etudes Vétérinaires et Alimentaires, Laboratoire des Médicaments Vétérinaires, Javené, 35133 Fougères, France
Downloaded by [Monash University Library] at 13:44 13 September 2013
(Received 26 May 1989; revised 12 January 1990; accepted 19 January 1990) An investigation was conducted into the elimination into milk of four sulphonamides (sulphadimidine, sulphamethoxypyridazine, sulphadimethoxine and sulphadoxine) after intravenous injections of drugs available in France. Sulphonamide determination in milk was performed using a high-performance liquid chromatography (HPLC) method with a detection limit of 0.01 µg/ml. These results were used to determine the withdrawal times (two to six milkings) required for these drugs with a tolerance of 0.1 µg/ml as proposed by the European countries. Keywords: sulphonamide, residues, milk, cows
Introduction
Before antibiotics were used in veterinary medicine, parenteral administration of sulphonamides was practically the only means of treating infectious diseases. Despite the discovery of penicillin and the widespread use of antibiotics, these antimicrobial agents have remained as popular, although the use of sulphonamide declined temporarily (Mol 1975). They are probably still the most commonly used antimicrobial agents throughout the world for the treatment of bovine pneumopathies because of their broad spectrum of activity and their low cost (Brugère et al. 1985). Many studies of their elimination into milk after therapeutic treatment of milking cows were conducted. Several methods for sulphonamide assays in milk were described: microbiological methods (Bogaerts etal. 1981), colorimetric methods based on the Bratton and Marshall reaction (Houston and Umstead 1968, Selzer and Banes 1963, Tishler et al. 1968), gas chromatography (Holtmannspotter and Thier 1982), liquid chromatography (Alawi and Rüssel 1981, Petz 1983, Soback et al. 1983, Nouws et al. 1985). The present study examines the elimination into milk of the main sulphonamides used in dairy cows: sulphadimidine (SDM), sulphamethoxypyridazine (SMP), sulphadimethoxine (SDT) and sulphadoxine (SDX) administered intravenously. The analytical method chosen, highperformance liquid chromatography (HPLC), was sensitive enough (0-01 fig/ml) to obtain sufficiently precise withdrawal times. Materials and methods
Animals The experiments were conducted on a herd of 14 dairy cows (Friesan or 0265-203X/90 $3.00 © 1990 Taylor & Francis Ltd.
528
Β. Roudaut and J. P. Moretain
Normande breeds) at different lactating stages. The animals were kept under usual breeding farm conditions and were milked by machine twice a day. The daily milk production ranged from 10 to 22 litres.
Downloaded by [Monash University Library] at 13:44 13 September 2013
Drugs The four drugs were respectively aqueous solution of sodium sulphadimidine (33g/100ml), sodium sulphamethoxypyridazine (25g/100ml), sodium sulphadimethoxine (25 g/100 ml) and sodium sulphadoxine (20 g/100 ml and 4 g/100 ml trimethoprim). They were administrered at the dosages indicated by the manufacturer after checking sulphonamide concentration. Treatment After the morning milking, drugs were administered to five cows by the intravenous route into the jugular vein using the recommended dose: 100 mg/kg for SDM, 75 mg/kg for SMP, 50 mg/kg for SDT, and 20 mg/kg for SDX. Milking The cows were milked twice a day at 8.00 a.m. and 5.00 p.m. For each cow, samples of 100 ml were taken from the total milk obtained from all quarters collected in individual milk cans after the treatment began. The samples were frozen pending analysis. Analysis Sulphonamides were determined by a liquid chromatography method after extraction of milk and clean-up. Apparatus and Chromatographie conditions. The HPLC system consisted of a SP 8700 pump (Spectra-Physics) coupled with a Spectroflow 773 (Kratos) variable UV detector, which was connected to an integrator (Shimadzu-CR IB). The injector was a Rheodyne 7125 with a 50 μΐ loop. The column was a Novapak C 18, 5 μΐη, 150 X 3-9 mm (Waters). The detection of sulphonamides was performed at 265 nm (SDM, SMP) and 271 nm (SDT, SDX). The mobile phase was acetonitrile, 0-01 M ammonium acetate solution pH 6-6 (10:90, v/v) and the solvent rate was 1 ml/min. Sample preparation. Five millilitres of raw milk, to which was added 20 μ\ HC1 12 Ν for SDT, homogenized by ultrasonication, were extracted with 25 ml ethyl acetate for 10 min (rotary shaker REAX 2, Heidolph). After centrifugation for 5 min at 1500 g, 20 ml of the ethyl acetate extract was evaporated to dryness with a rotary evaporator (Büchi). The dry extract was dissolved in 10 ml HC1 Ν and transferred to a centrifuge tube. The aqueous phase was rinsed with 10 ml of méthylène chloride (or hexane for SDT and SDX) adjusted to pH 5 · 5 with 0 · 9 ml of 10 Ν NaOH and 5 ml of 1 M monobasic potassium phosphate (pH 6*0) then extracted twice with 10 ml of dichloromethane. The organic layer was evaporated to dryness with a rotary evaporator and dissolved in 2 ml of the mobile phase. A 50 μΐ aliquot of the solution was injected for HPLC analysis. The analytical technique used led to detection of concentrations
