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Resumen El objetivo del presente trabajo fue determinar el porcentaje de bovinos para abasto positivos a clorhidrato de clembuterol (CCL), con base en muestras de orina de bovinos sacrificados en 3 rastros del estado de México. Para esto se colectaron muestras en los rastros municipales de Toluca (59), Ixtlahuaca (52) y Atlacomulco (40), y se obtuvo un total de 151 muestras. El análisis de las muestras se realizó a través de la prueba de ELISA y para el reporte de resultados se utilizó estadística descriptiva. De las muestras analizadas 105 fueron positivas (69,53 %), por rastro la positividad, fue de 48, 30 y 27 respectivamente. Al establecer diferentes rangos de concentración a CCL, en el rango de 200 a 1999 pg g-1 se ubicaron 46 muestras; 6 muestras, en el rango de 2000 a 3525 pg g-1; 3, en el rango de 3526 a 5050 pg g-1; 1 muestra, entre 5051 y 6575 pg g-1, y 95 se ubicaron entre 6575 y >8100 pg g-1. El rastro con mayor porcentaje de positividad fue el de Toluca, con base en los rangos establecidos, y las concentraciones más elevadas también se observaron en el mismo rastro. El límite mínimo de detección de CCL de 2000 pg g-1 permite considerar que, con base tanto en la concentración como en su cinética, las cantidades detectadas en el estudio sirven como un indicador importante de residualidad a través de este tipo de muestra.
Abstract The present work aimed to determine the percentage of bovines that tested positive for clenbuterol hydrochloride (CCL), based on urine samples of cattle slaughtered in 3 slaughterhouses of the State of Mexico. To the effect, samples were collected in the municipal slaughterhouses of Toluca (59), Ixtlahuaca (52), and Atlacomulco (40), resulting in a total of 151 samples. Sample analysis was carried out using the ELISA test, and descriptive statistics were used to report the results. 105 of the analyzed samples were positive (69.53%); by slaughterhouse, positivity was 48, 30, and 27, respectively. When establishing different ranges of CCL concentration, 46 samples were in the range of 200 to 1999 pg g-1; 6 samples were in the range of 2000 to 3525 pg g-1; 3 were in the range of 3526 to 5050 pg g-1; 1 sample was between 5051 and 6575 pg g-1, and 95 samples were between 6575 and >8100 pg g-1. The slaughterhouse with the highest percentage of positivity was that of Toluca, based on the established ranges, and the highest concentrations were also observed there. The minimum limit for the detection of CCL (2000 pg g-1) allows considering that, based on both concentration and kinetics, the quantities detected in the study serve as an important indicator of residuality in this type of sample.
Resumo O objetivo do presente trabalho foi determinar a porcentagem de bovinos para fornecimento, positivos ao cloridrato de Clenbuterol (CCL), baseados em amostras de urina de bovinos abatidos em três trilhas do estado de México. Para isso, foram coletadas amostras nas trilhas municipais de Toluca (59), Ixtlahuaca (52) e Atlacomulco (40), e obtivera-se um total de 151 amostras. A análise das amostras realizou-se através do teste de ELISA e para o relatório de resultados foi usada estatística descritiva. Das amostras analisadas 105 foram positivas (69,53 %), pelo traço a positividade foi de 48, 30 e 27 respetivamente. Ao estabelecer diferentes faixas de concentração a CCL, 46 amostras foram colocadas na faixa de 200 a 1999 pg g-1; 6 amostras na faixa de 2000 a 3525 pg g-1; 3 na faixa de 3526 a 5050 pg g-1; 1 amostra entre 5051 e 6575 pg g-1, e 95 entre 6575 e >8100 pg g-1. A trilha com maior porcentagem de positividade foi a de Toluca, baseada nas faixas estabelecidas e as concentrações mais altas observaram-se na mesma trilha. O limite mínimo de detecção de CCL de 2000 pg g-1 permite considerar que, baseado na concentração e na cinética, as quantidades detectadas no estudo servem como indicador importante de residualidade através deste tipo de amostra.
ABSTRACT
Objective To establish a high performance liquid chromatography (HPLC) through the optimization of the chromatographic conditions, which can detect the contents of clenbuterol hydrochloride (CL) residues in animal edible product in a large quantity. Methods The animal edible product were extracted by perchloric acid, and then impurities were removed by liquid-liquid extraction (LLE) which used ethyl acetate- isopropanol. After the organic phase was concentrated, C18 column (150 mm×4.6 mm, 5 μm) was used to separate CL. Mobile phase were methanol-sodium dihydrogen phosphate, and then determined by HPLC. Results A good linear response was obtained over the range of 0.2-10.0 μg/mL with the correlation coefficient (r) 0.99984. The method determination limit was 0.15 μg/kg which was lower than the National standard method 0.5μg/kg. The retention time of the CL was 6.51 min, the chromatographic peak was good. The recovery rates spiked with standards 1.6-12 μg were 92.86％-100.93％, which was higher than National standard method (89.79％-92.36％) . The precision of intra-day and inter-day were both under 5％, which lower than National standard. Conclusion The optimized chromatographic conditions are suitable for the large quantity determination of clenbuterol hydrochloride in animal edible product.
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Resumen El clorhidrato de clembuterol (CCL) es un p-agonista promotor del crecimiento en animales para abasto, pero su uso ilícito ha generado repercusiones en salud pública. Se realizó un modelo biológico con ratones, con el objeto de evaluar el efecto del CCL sobre la ganancia de peso y las lesiones histológicas que ocasiona. Los ratones fueron alimentados con carne de conejo, que previamente fue suplementada con CCL. Treinta y cinco días posexposición se registró el peso corporal; se obtuvo la concentración muscular y sérica de CCL a través de la prueba de ELISA, y se colectaron tejidos (hígado y corazón) para análisis histopatológico. Los valores obtenidos de los animales experimentales (G1 y G2) se analizaron mediante un diseño experimental completamente al azar con dos tratamientos (n = 10), sometidos a un análisis de varianza y comparación de medias con la prueba de Tukey (p < 0,05). Se registró un incremento de peso corporal de 7 g en el G1, contra 4,0 g del G2. El peso del hígado fue de 2,58 g y 1,79, respectivamente (p < 0,05). En el G1 la concentración muscular de CCL fue 5324 pg g-1 y en suero sanguíneo de 4378 pg g-1. Solo se observaron cambios histológicos en tejidos de los ratones del G1. El hígado mostró tumefacción celular, mitosis moderada, picnosis y degeneración hidrópica; en corazón, engrosamiento de fibras, pleomorfismo e hileración nuclear. El CCL favoreció el incremento de peso en los ratones expuestos, y provocó alteraciones estructurales en hígado y corazón.
Abstract Clenbuterol hydrochloride (CLB) is a growth-promoting p-agonist in animals for supply, but its illicit use has generated repercussions on public health. A biological model with mice was developed to evaluate the effect of CLB on weight gain and histological lesions. Mice were fed rabbit meat, which was previously supplemented with CLB. Body weight was recorded 35 days post-exposure; muscular and serum concentration of CLB was obtained through the ELISA test, and tissues were collected from liver and heart for histopathological analysis. Values obtained from the experimental animals (G1 and G2) were analyzed by a completely randomized experimental design with two treatments (n = 10), subjected to an analysis of variance and comparison of means with the Tukey test (p<0.05). There was an increase of 7 g in body weight in G1, compared to 4.0 g in G2. Liver weight was 2.58 g and 1.79, respectively (p<0.05). In G1, CLB concentration in muscle was 5324 pg.g-1, and 4378 pg.g-1 in blood serum. Only histological changes were observed in the tissues of Gl mice. Liver showed cellular swelling, moderate mitosis, pyknosis and hydropic degeneration; in addition, fiber thickening, pleomorphism and nuclear atypia were observed in the heart. CLB contributed to weight gain in exposed mice and caused structural alterations in liver and heart.
Resumo O cloridrato de clembuterol (CCL) é um b-agonista promotor do crescimento em animais para abate, porém o seu uso ilícito tem gerado repercussões em saúde pública. Fezse um modelo biológico com ratos, com o objeto de avaliar o efeito do CCL sobre o ganho de peso e as lesões histológicas que ocasiona. Os ratos foram alimentados com carne de coelho, que previamente foi suplementada com CCL. Trinta e cinco dias pós-exposição se registrou o peso corporal; obteve-se a concentração muscular e sérica de CCL através da prova de ELISA, e foram coletados tecidos (fígado e coração) para análise histopatológica. Os valores obtidos dos animais experimentais (Gl e G2) foram analisados mediante um desenho experimental completamente ao acaso com dois tratamentos (n = 10), submetidos a uma análise de variações e comparação de médias com a prova de Tukey (p < 0,05). Registrou-se um aumento de peso corporal de 7 g no G1, contra 4,0 g do G2. O peso do fígado foi de 2,58 g e 1,79, respectivamente (p < 0,05). No G1 a concentração muscular de CCL foi 5324 pg g-1 e em soro sanguíneo de 4378 pg g-1. Somente foram observadas mudanças histológicas em tecidos dos ratos do G1. O fígado apresentou inchaço celular, mitose moderada, picnose e degeneração hidrópica; no coração, engrossamento de fibras, pleomorfismo e filamento nuclear. O CCL favoreceu o aumento de peso nos ratos expostos, e provocou alterações estruturais em fígado e coração.
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Objective: To establish a rapid and accurate method for the determination of 15 chemical drugs which were illegally added into the slimming Chinese patent medicines (CPM) and health foods. Methods: The UPLC-MS/MS method was adopted. The samples were extracted with methanol by ultrasonic processing and separated on a Waters Acquity BEH C18 column (100 mm × 2.1 mm, 1.7 μm) with 0.1% formic acid methanol (A) -0.1% formic acid water (B) as mobile phase by gradient elution (0-3 min, 33%-45% A; 3-5 min, 45%-55% A; 5-7 min, 55%-70% A; 7-9 min, 70%-80% A; 9-10 min, 80%-90% A; 10-11 min, 90%-33% A; 11-13 min, 33% A at a flow rate of 0.2 mL/min, and the column temperature was 40℃. A positive-ion (ESI+) source and an MRM mode were used to separate and quantitatively determine 15 chemical drugs. The obtained molecular ions, fragment ions, and retention time for MRM channels were used to identify the 15 kinds of drugs by comparison with those of reference substances. The obtained peak areas were used to determine the accurate contents of chemical drugs in CPM and the health foods. Results: A good resolution of 15 kinds of chemical drugs, including terbutaline hydrochloride, ephedrine hydrochloride, theophylline, caffeine, doxofylline, clenbuterol hydrochloride, tulobuterol hydrochloride, bambuterol hydrochloride, fenfluramine hydrochloride, furosemide, indapamide, phenolphthalein, sibutramine hydrochloride, N-demethylated sibutramine hydrochloride, and hydrochloric acid N,N-dinor sibutraminel, was obtained under this UPLC and MS/MS condition. The limits of qualitation and quantitation were in the range of 0.1-5.0 ng/g and 0.3-15.0 ng/g. The standard addition recoveries were in the range of 91.8%-110.8%. In the 86 batches of samples (including capsules, granules, and other different matrix types) were detected in the 74 batches of added chemicals, the positive rate was 86.0%. Sibutramine hydrochloride (39 batches), furosemide (20 batches), phenolphthalein (23 batches), theophylline (1 batch), and caffeine (15 batches) were checked out in the samples, 22 batches of which two kinds were checked out, one batch of which three kinds were checked out. By contrast, the products which were not clearly marked manufacturer illegally added more seriously. Conclusion: The method is simple, accurate, and highly sensitive, which can be used for the determination of illegally added chemical drugs in slimming CPM and health foods.
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En este trabajo se analiza y valora la información sobre las características y riesgos del uso de clorhidrato de clembuterol (CCL) por su potencial efecto tóxico, debido a su inclusión en la alimentación animal (bovinos, cerdos, ovinos y aves) con la finalidad de mejorar parámetros productivos-reproductivos, pero que descuidan la inocuidad alimenticia. Por eso es necesario revalorar los peligros potenciales que puede ocasionar al emplearse tanto en medicina humana como en veterinaria. El CCL es un (β-adrenérgico sintético, polvo blanco, anhidro, muy soluble en agua y altamente estable a temperatura ambiente. Es utilizado en forma clandestina en animales de engorde destinados al consumo humano. Terapéuticamente, se utiliza como un medicamento broncodilatador (pacientes con asma); en competencias deportivas se ha detectado su uso ilegal (dopaje), y en fisicoculturismo es utilizado por su efecto anabólico. Su administración en el ganado para abasto modifica e incrementa el crecimiento de masas musculares, y disminuye la acumulación de grasa; se acumula en diferentes órganos. En personas con antecedente del consumo de hígado de bovino contaminado con CCL se ha registrado temblor, dolor muscular, mareos, cefalea y taquicardia. En México, de manera ilegal y clandestina, hay distribución, comercialización y uso de CCL; sin embargo, el trabajo de las organizaciones ganaderas para el registro de unidades de producción libres de esta sustancia garantizará el consumo de los productos cárnicos. Proponer el uso de otras sustancias de las cuales hasta el momento no se tiene indicios de toxicidad propiciará una productividad sustentable, segura e inocua en las unidades de producción pecuaria.
This paper analyzes and evaluates information about the characteristics and risks of using Clenbuterol hydrochloride (CCL) for their potential toxic effects, due to its inclusion in animal food (cattle, pigs, sheep and poultry) to improve productive-reproductive parameters, but neglecting food safety. Therefore, it is necessary to reassess the potential dangers that may result when used in both human and veterinary medicine. The (β-adrenergic synthetic CCL, white powder, anhydrous, highly water soluble and highly stable at room temperature is used in a clandestine manner to fatten animals for human consumption. Therapeutically, it is used as a bronchodilator drug (asthma patients); its illegal use (doping) has been detected in sports competitions, and it is used for bodybuilding due to its anabolic effect. Its use in cattle for slaughter modifies and increases the growth of muscle mass and reduces fat accumulation, which accumulates in different organs. In people with a history of bovine liver consumption contaminated with CCL, there has been registration of: tremor, muscle pain, dizziness, headache, and tachycardia. In Mexico, in an illegal and clandestine manner, there is distribution, marketing and use of CCL; however, the work of livestock organizations in registering production units free of this substance will ensure the consumption of meat products. Moreover, to propose the use of other substances which so far have no signs of toxicity will lead to a sustainable, secure and safe productivity in livestock units.
Neste trabalho se analisa e avalia a informação sobre as características e riscos do uso de cloridrato de clembuterol (CCL) pelo seu potencial efeito tóxico, devido à sua inclusão na alimentação animal (bovinos, cerdos, ovinos e aves) com a finalidade de melhorar parâmetros produtivo-reprodutivos, mas que descuidam a inocuidade alimentícia. Por isso é necessário reavaliar os perigos potenciais que pode ocasionar ao empregar-se tanto em medicina humana como veterinária. O CCL β-adrenérgico sintético, pó branco, anidro, muito solúvel em água e altamente estável a temperatura ambiente. É utilizado em forma clandestina em animais de corte destinados ao consumo humano. Terapeuticamente, se utiliza como um medicamento bronco dilatador (pacientes com asma); em competências esportivas tem sido detectado o seu uso ilegal (dopagem), e em físico culturismo é utilizado por seu efeito anabólico. Sua administração no gado para abasto modifica e incrementa o crescimento de massas musculares, e diminui a acumulação de gordura; acumulando-se em diferentes órgãos. Em pessoas com antecedente do consumo de fígado de bovino contaminado com CCL tem se registrado: tremor, dor muscular, tonturas, cefaleia e taquicardia. No México, de maneira ilegal e clandestina, há distribuição, comercialização e uso de CCL; contudo, o trabalho das organizações pecuaristas para o registro de unidades de produção livres desta substância garantirá o consumo dos produtos de carne. Assim como também, propor o uso de outras substâncias das quais até o momento não se tem indícios de toxicidade propiciará uma produtividade sustentável, segura e inócua nas unidades de produção pecuária.
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Objective To evaluate the clinical curative effect of ammonia bromine,oral solution (Yitanjing) auxiliary for the treatment of children with bronchopneumonia.Methods The clinical data of 64 children with acute bronchopneumonia with breathing were retrospectively analyzed.According to different treatment,they were divided into three groups,three groups were intravenously given anti-infective drug therapy,group A (22 cases) treated with Yitanjing,B group (21 cases) treated with ammonia bromine rope,group C (21 cases) treated with hydrochloric acid C quattro.Before and after the treatment,cough,sputum improve scores,and the changes of pulmonary function,hospitalization time were observed.The clinical efficacy of three groups of children with and medication compliance were evaluated.Results In group A,group B and group C,the effective rates were 95.45％,90.48％ and 95.45％ respectively,the effective rate of group A was significantly higher than group B and group C (x2 =3.88,4.27,all P ＜ 0.05) ;In group A good taste, convenient selected rates were 95.45％,95.45％,which were significantly higher than 85.71％,90.48％ of group B (x2 =4.17,3.86,all P ＜ 0.05) and C group (47.62％,71.73％) (x2 =8.67,6.86,all P ＜ 0.05).Conclusion Ammonia bromine,oral liquid can effectively improve pulmonary function in children with acute bronchial pneumonia and clinical symptoms such as cough,sputum,high compliance.
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Objective To observe the effect of montelukast sodium and ambroxol oral solution in treatment respiratory symptoms caused by mycoplasma infection in children.Methods 147 children who were infected by mycoplasma pneumonia with lower respiratory tracts symptom were randomly divided into A,B,C three groups:group A was control group,group B was treated by montelukast sodium,and group C was treated by montelukast sodium and ambroxol oral solution.Coughing,symptomatic relief of pant were observed in each group.Results After 7 days treatment,the total effective rate of three groups was A group 79.5％,B group 95.7％,C group 96.4％.Compared with group A,the coughing and panting time in group B and group C were significantly shorter(x2 =5.61,9.54,all P ＜0.05),The rate of coughing and panting complete remission in group C was higher compared with group B (x2 =5.39,P ＜ 0.05),but there was no statistificantly significant difference in the total efficency between the two groups.Conclusion Montelukast sodium and ambroxol oral solution can effectively improve the symptoms of respiratory tract infection caused by mycoplasma pneumoniae.