喹赛多在猪、鸡和鲤可食性组织中的残留消除研究
发布时间：2018-09-07 16:24
【摘要】：喹赛多作为喹嗯啉类新药,对畜禽及水产品中常见病原菌有良好的抑菌作用,并且能够提高饲料转化率,促进机体生长。与同类药物相比,喹赛多具有安全性更高、毒副作用更低的特点。而作为食品动物用药,对其安全性评价是新药开发的重要内容。兽药残留是影响食品安全的主要因素,但是关于喹赛多在畜禽及水产品中的残留消除研究并没有完全清楚。目前国际上公认的可食性组织包括肝、肾、肌肉、脂肪,而在我国,除了肝、肾、肌肉和脂肪外,主要的可食性组织还有心、肺、胃、大肠和小肠。目前关于喹赛多的残留消除研究已在畜禽的肝、肾、肌肉、脂肪中开展,但是喹赛多在心、肺、胃、大肠和小肠中的残留消除研究仍未见报道。本课题以猪、鸡和鲤鱼为研究对象,建立了喹赛多及其5种主要代谢产物在各可食性组织中残留检测的高效液相色谱方法,并对这些化合物的残留消除规律进行研究。本研究表明,喹赛多用于猪、鸡和鲤之后,吸收和消除迅速。通过本研究进一步确定了喹赛多在猪和鸡体内的残留靶组织是肾脏,残留标示物是Cyl；在鲤体内的残留靶组织是胃肠,残留标示物是Cy1。阐明了喹赛多在动物体内的安全性,为食品安全评价提供理论依据和技术支持。1残留检测方法的建立本实验建立了喹赛多(Cy0)、脱二氧喹赛多(Cyl)、N4-脱一氧喹赛多(Cy2)、N-(喹VA啉-2-甲基)-氰乙酰肼(Cy4)、喹嗯啉-2-羧酸(Cy6)和2-氰基-N'-(N1-氧喹VA啉-2-亚甲基)乙酰肼(Cy9)六种化合物在猪和鸡的肝脏、肾脏、肌肉、脂肪、心、肺、胃、大肠和小肠9种可食性组织：鲤鱼的肝脏、肾脏、肌肉、脂肪、皮肤和胃肠道6种可食性组织中的残留检测方法。各组织依次通过乙酸乙酯提取、体积比为1：50：10：39的甲酸：甲醇：乙腈：水复合试剂提取、正己烷去脂和亲水亲脂平衡型共聚物(HLB)小柱净化,最后经体积比为15：85的乙腈：水混合试剂复溶,320 nm紫外波长下通过高效液相色谱进行检测。该方法下Cy0、Cy1和Cy6在猪和鸡的肌肉、心脏、肺脏及鱼的肌肉、皮肤和胃肠的定量限为15μg/kg,在猪和鸡的肝、肾、脂肪、胃、大肠和小肠及鱼肝、肾和脂肪中的定量限为20μg/kg;Cy2、Cy4和Cy9在猪和鸡的肌肉、心脏、肺脏及鱼的肌肉、皮肤和胃肠的定量限为20μg/kg,在猪和鸡的肝、肾、脂肪、胃、大肠和小肠及鱼肝、肾和脂肪中的定量限为40μg/kg。添加不同定量限水平上浓度的药物,回收率在均在60%以上,且日间变异系数在12%以下。农业部规定残留检测方法日间相对标准偏差17%-21%,本实验建立的方法满足该要求,能用于喹赛多及其代谢产物的定量分析。2喹赛多在猪、鸡和鲤体内的残留消除研究健康三元杂交猪25头、20日龄科宝肉鸡30羽和健康鲤鱼30尾,各分成5组。连续饲喂添加200 mg/kg喹赛多的饲料2周,停药后于不同时间点宰杀一组,收肝脏、肾脏、肌肉、脂肪、心、肺、胃、大肠和小肠。按上述方法对组织中药物的含量进行测定,结果如下：猪：停药6 h,除脂肪外各组织均检测到原形和QCA,药物浓度范围分别为21.8-66.3μg/kg和22.4-56.9μg/kg;各组织中均检测出Cyl,浓度范围是20.4-66.0μg/kg,原形和QCA在猪体内消除迅速。停药1 d,各组织中原形均消除至定量限以下,肾脏和肺脏中检测出QCA,浓度分别为22.3 μg/kg和25.1 μg/kg;肝、肾、肺和小肠中检测到Cyl,浓度范围是20.8-44.2μg/kg,肾脏最高。停药3 d,各组织中QCA均消除至定量限以下,只有肾脏中能检测到少量的Cyl。停药7d各组织中Cyl均低于定量限。鸡：肌肉和脂肪没有检测到任何代谢产物。停药6 h,原形药物只在肝脏、肾脏、小肠、心、肺和胃中检测出,浓度范围是21.4-67.2μg/kg,肾脏最高；肝脏、肾脏、心脏、肺脏和小肠中均检测到QCA,浓度范围为24.7-66.8μg/kg,肾脏中最高；肝脏、肾脏、心、肺、大肠和小肠检测出Cyl,浓度范围是32.8-65.6μg/kg。停药1 d,原形和QCA仅在肾脏中检测出,浓度分别为34.1和32.5μg/kg;在肝、肾和肺中检测出Cyl,浓度分别为32.0、47.4和23.0μg/kg。停药3 d,仅在肾中检测出少量的Cy1。停药5 d,各组织中检测不到任何代谢产物。鲤：停药6 h,各组织中除原形、脱二氧喹赛多和QCA外,还在皮肤和胃肠中检测出Cy4,在肾脏、肌肉、脂肪、皮肤和胃肠中检测出原形,浓度范围为15.2-25.5 μg/kg肾脏居多；肝、肾、皮肤和胃肠中检测到QCA,浓度范围为20.2-36.1μg/kg各组织中均检测到Cyl,浓度范围为32.4-40.8μg/kg,胃肠最高。停药1 d,各组织中的原形和QCA均消除至定量限以下；除脂肪外各组织中均检测到Cy1,浓度范围20.5-36.8μg/kg,胃肠最高。停药3 d,仅胃肠道有少量的Cy1。停药7d各组织中检测不到任何代谢产物。综上所述,本实验首次建立喹赛多及其5种主要主要代谢产物在猪和鸡9种可食性组织,鱼6种可食性组织中的残留检测方法,同时,首次阐明了喹赛多及其代谢产物在这些组织中的消除规律,其研究结果为指导临床合理用药、药物安全性评价和残留标示物的确定提供了科学依据。
[Abstract]:As a new quinoxaline drug, quinoxaldol has a good bacteriostatic effect on common pathogenic bacteria in livestock, poultry and aquatic products, and can improve feed conversion rate and promote the growth of the body. Compared with the same drugs, quinoxaldol has higher safety and lower side effects. As a food animal drug, its safety evaluation is a new drug development. Veterinary drug residue is a major factor affecting food safety, but the study on the elimination of quinacetin residue in livestock, poultry and aquatic products is not entirely clear. Lung, stomach, large intestine and small intestine. At present, the study on the elimination of quetiapine has been carried out in the liver, kidney, muscle and fat of livestock and poultry. However, the study on the residual elimination of quinazadiam in heart, lung, stomach, large intestine and small intestine has not been reported. In this study, quinazo and its 5 main metabolites were found in pigs, chickens and carps. The residues in the tissues were detected by high performance liquid chromatography, and the residue elimination rule of these compounds was studied. This study shows that quetiapine can rapidly absorb and eliminate after being used in pigs, chickens and carps. Through this study, it is further confirmed that the residual target tissue of quetial in pigs and chickens is the kidneys, and the residual marker is Cyl. The residual target tissue in vivo is the gastrointestinal tract, and the residual marker is Cy1. The safety of quinacetin in vivo was clarified, which provides theoretical basis and technical support for food safety evaluation. 1 The method of residue detection was established. Cy0, Cyl, N4-deoxyquinacetin (Cy2), N-(quinoVAline-2-methyl)-cyanoethyl. Hydrazine (Cy4), quinoline -2- carboxylic acid (Cy6) and 2- cyano -N'- (N1- oxoquine VA VA -2- Ya Jiaji) acetyl hydrazine (Cy9) six compounds in pig and chicken liver, kidney, muscle, fat, heart, lung, stomach, large intestine and small intestine 9 kinds of edible tissue: carp liver, kidney, muscle, fat, skin and gastrointestinal 6 kinds of edible tissue residues detection method. The tissues were extracted by ethyl acetate with a volume ratio of 1:50:10:39, formic acid: methanol: acetonitrile: water complex reagent extraction, n-hexane degreasing and hydrophilic lipophilic equilibrium copolymer (HLB) column purification, and finally re-dissolved by acetonitrile: water mixture reagent with a volume ratio of 15:85, and carried out by high performance liquid chromatography at 320 nm ultraviolet wavelength. The limit of quantification for Cy0, Cy1 and Cy6 in pigs, chickens * muscles, heart, lungs and fish muscle, skin and gastrointestinal was 15 * g/kg, and the limit of quantification in pigs, chickens, liver, kidney, fat, stomach, large intestine and small intestine and liver, kidney and fat was 20 * g/kg; Cy2, Cy4 and Cy9 in muscle, heart, lungs and fish muscles, skin and stomach of pigs and chickens. The limit of quantification was 20 * g/kg, and the quantitation limit for liver, kidney, fat, stomach, large intestine, small intestine, liver, kidney and fat of pigs and chickens was 40 g/kg.. The recoveries were over 60% and the coefficient of variation of day was below 12% when adding different quantitation limits. The Ministry of agriculture stipulated that the relative standard deviation of residue detection method was 17%-21%. The established method can meet the requirement and can be used for the quantitative analysis of quetiapine and its metabolites. The elimination of.2 * quetiapine residues in pigs, chickens and carp * three healthy 25 crossbred pigs, 30 days of 20 days old Cobb broilers and 30 healthy carp Cyprinus carpio were divided into 5 groups. Continuous feeding of 200 mg/kg quetiapine feed for 2 weeks, after withdrawal of different drugs. At the time point, a group was slaughtered, and liver, kidney, muscle, fat, heart, lung, stomach, large intestine and small intestine were collected. * the contents of the drugs in the tissues were determined according to the above methods. The results were as follows: pigs were stopped for 6 h, and the original and QCA were detected in all tissues except fat. The concentrations of drugs ranged from 21.8-66.3 to g/kg and 22.4-56.9 g/kg respectively, and C was detected in all tissues. YL, the concentration range was 20.4-66.0 * g/kg, the original shape and QCA were eliminated rapidly in pigs. After stopping 1 D, the prototypes of all tissues were removed below the quantitation limit, and QCA in the kidneys and lungs was detected. The concentrations were 22.3 g/kg and 25.1 g/kg respectively. Cyl was detected in the liver, kidney, lung and small intestine, the concentration range was 20.8-44.2, g/kg, the kidney was the highest, the drug withdrawal was 3, and the tissues were different. Chicken: No metabolites were detected in muscle and fat. Within 6 hours, prototype drugs were detected only in liver, kidney, small intestine, heart, lung and stomach. The concentration range was 21.4-67.2 ug/kg, the highest in kidney. QCA was detected in kidney, heart, lung and small intestine, the concentration range was 24.7-66.8 ug/kg, the highest in kidney; Cyl was detected in liver, kidney, heart, lung, large intestine and small intestine, the concentration range was 32.8-65.6 ug/kg. Cy4 was detected in the skin and gastrointestinal tract, besides prototype, deoxyquinocetone and QCA. Cy4 was detected in the kidney, muscle, fat, skin and gastrointestinal tract. QCA was detected in liver, kidney, skin and gastrointestinal tract. Cyl was detected in 20.2-36.1 ug/kg tissues. The concentration ranged from 32.4-40.8 ug/kg, and the gastrointestinal tract was the highest. 36.8 g/kg, the highest gastrointestinal tract. 3 D was stopped. Only a small amount of Cy1. was stopped in the gastrointestinal tract. NO metabolites were detected in all tissues of 7D. In summary, this experiment established for the first time that quinazo and its 5 main metabolites * 9 kinds of edible tissues in pigs and chickens, and residue detection methods in 6 edible tissues. The elimination of polymorphisms and their metabolites in these tissues provides a scientific basis for clinical rational drug use, drug safety evaluation and determination of residual markers.
【学位授予单位】：华中农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S859.84
，
本文编号：2228786
[Abstract]:As a new quinoxaline drug, quinoxaldol has a good bacteriostatic effect on common pathogenic bacteria in livestock, poultry and aquatic products, and can improve feed conversion rate and promote the growth of the body. Compared with the same drugs, quinoxaldol has higher safety and lower side effects. As a food animal drug, its safety evaluation is a new drug development. Veterinary drug residue is a major factor affecting food safety, but the study on the elimination of quinacetin residue in livestock, poultry and aquatic products is not entirely clear. Lung, stomach, large intestine and small intestine. At present, the study on the elimination of quetiapine has been carried out in the liver, kidney, muscle and fat of livestock and poultry. However, the study on the residual elimination of quinazadiam in heart, lung, stomach, large intestine and small intestine has not been reported. In this study, quinazo and its 5 main metabolites were found in pigs, chickens and carps. The residues in the tissues were detected by high performance liquid chromatography, and the residue elimination rule of these compounds was studied. This study shows that quetiapine can rapidly absorb and eliminate after being used in pigs, chickens and carps. Through this study, it is further confirmed that the residual target tissue of quetial in pigs and chickens is the kidneys, and the residual marker is Cyl. The residual target tissue in vivo is the gastrointestinal tract, and the residual marker is Cy1. The safety of quinacetin in vivo was clarified, which provides theoretical basis and technical support for food safety evaluation. 1 The method of residue detection was established. Cy0, Cyl, N4-deoxyquinacetin (Cy2), N-(quinoVAline-2-methyl)-cyanoethyl. Hydrazine (Cy4), quinoline -2- carboxylic acid (Cy6) and 2- cyano -N'- (N1- oxoquine VA VA -2- Ya Jiaji) acetyl hydrazine (Cy9) six compounds in pig and chicken liver, kidney, muscle, fat, heart, lung, stomach, large intestine and small intestine 9 kinds of edible tissue: carp liver, kidney, muscle, fat, skin and gastrointestinal 6 kinds of edible tissue residues detection method. The tissues were extracted by ethyl acetate with a volume ratio of 1:50:10:39, formic acid: methanol: acetonitrile: water complex reagent extraction, n-hexane degreasing and hydrophilic lipophilic equilibrium copolymer (HLB) column purification, and finally re-dissolved by acetonitrile: water mixture reagent with a volume ratio of 15:85, and carried out by high performance liquid chromatography at 320 nm ultraviolet wavelength. The limit of quantification for Cy0, Cy1 and Cy6 in pigs, chickens * muscles, heart, lungs and fish muscle, skin and gastrointestinal was 15 * g/kg, and the limit of quantification in pigs, chickens, liver, kidney, fat, stomach, large intestine and small intestine and liver, kidney and fat was 20 * g/kg; Cy2, Cy4 and Cy9 in muscle, heart, lungs and fish muscles, skin and stomach of pigs and chickens. The limit of quantification was 20 * g/kg, and the quantitation limit for liver, kidney, fat, stomach, large intestine, small intestine, liver, kidney and fat of pigs and chickens was 40 g/kg.. The recoveries were over 60% and the coefficient of variation of day was below 12% when adding different quantitation limits. The Ministry of agriculture stipulated that the relative standard deviation of residue detection method was 17%-21%. The established method can meet the requirement and can be used for the quantitative analysis of quetiapine and its metabolites. The elimination of.2 * quetiapine residues in pigs, chickens and carp * three healthy 25 crossbred pigs, 30 days of 20 days old Cobb broilers and 30 healthy carp Cyprinus carpio were divided into 5 groups. Continuous feeding of 200 mg/kg quetiapine feed for 2 weeks, after withdrawal of different drugs. At the time point, a group was slaughtered, and liver, kidney, muscle, fat, heart, lung, stomach, large intestine and small intestine were collected. * the contents of the drugs in the tissues were determined according to the above methods. The results were as follows: pigs were stopped for 6 h, and the original and QCA were detected in all tissues except fat. The concentrations of drugs ranged from 21.8-66.3 to g/kg and 22.4-56.9 g/kg respectively, and C was detected in all tissues. YL, the concentration range was 20.4-66.0 * g/kg, the original shape and QCA were eliminated rapidly in pigs. After stopping 1 D, the prototypes of all tissues were removed below the quantitation limit, and QCA in the kidneys and lungs was detected. The concentrations were 22.3 g/kg and 25.1 g/kg respectively. Cyl was detected in the liver, kidney, lung and small intestine, the concentration range was 20.8-44.2, g/kg, the kidney was the highest, the drug withdrawal was 3, and the tissues were different. Chicken: No metabolites were detected in muscle and fat. Within 6 hours, prototype drugs were detected only in liver, kidney, small intestine, heart, lung and stomach. The concentration range was 21.4-67.2 ug/kg, the highest in kidney. QCA was detected in kidney, heart, lung and small intestine, the concentration range was 24.7-66.8 ug/kg, the highest in kidney; Cyl was detected in liver, kidney, heart, lung, large intestine and small intestine, the concentration range was 32.8-65.6 ug/kg. Cy4 was detected in the skin and gastrointestinal tract, besides prototype, deoxyquinocetone and QCA. Cy4 was detected in the kidney, muscle, fat, skin and gastrointestinal tract. QCA was detected in liver, kidney, skin and gastrointestinal tract. Cyl was detected in 20.2-36.1 ug/kg tissues. The concentration ranged from 32.4-40.8 ug/kg, and the gastrointestinal tract was the highest. 36.8 g/kg, the highest gastrointestinal tract. 3 D was stopped. Only a small amount of Cy1. was stopped in the gastrointestinal tract. NO metabolites were detected in all tissues of 7D. In summary, this experiment established for the first time that quinazo and its 5 main metabolites * 9 kinds of edible tissues in pigs and chickens, and residue detection methods in 6 edible tissues. The elimination of polymorphisms and their metabolites in these tissues provides a scientific basis for clinical rational drug use, drug safety evaluation and determination of residual markers.
【学位授予单位】：华中农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S859.84
，
本文编号：2228786
本文链接：https://www.wllwen.com/yixuelunwen/dongwuyixue/2228786.html
