脱氢乙酸钠在肉鸡组织中的残留消除

研究脱氢乙酸钠作为饲料防霉剂在鸡组织中的残留消除,为其在鸡组织中的残留监控及休药期的制定提供理论依据。黄羽肉鸡连续饲喂200 mg/kg脱氢乙酸钠30 d后,于不同时间分别取肌肉、肝脏、肾脏和皮脂样品,采用反相高效液相色谱(HPLC)方法测定样品中脱氢乙酸钠含量。休药1 d时,鸡肾脏、肝脏、肌肉和皮脂中脱氢乙酸钠残留量分别为0.68、0.59、0.34和0.20 mg/kg。随着休药时间延长,各组织中脱氢乙酯钠的残留量逐渐降低,脱氢乙酸钠在肉鸡各组织中的残留水平依次为肾脏、肝脏、肌肉和皮脂。在组织中的残留消除半衰期分别为肾脏5.86 d、肝脏6.02 d、肌肉8.88 d及皮脂为10.38 d。若以0.2 mg/kg为临时最大残留限量(MRLs),则脱氢乙酸钠在鸡不同组织中的休药期分别为肾脏12.37d、肌肉9.37 d、肝脏9.18 d、皮脂4.16 d。得出结论:200 mg/kg脱氢乙酸钠拌料饲喂肉鸡30 d,在肉鸡各组织中的残留水平较低,均低于0.7 mg/kg;残留消除相对较快。     

乙酰甲喹在斑马鱼中的代谢研究

为了阐明乙酰甲喹在水产动物中的代谢过程,本试验采用超高效液相色谱串联四级杆/飞行时间质谱（UPLC-Q/TOF-MS）技术及代谢软件Metabolynx^XS自动分析采集功能,研究其在斑马鱼中的主要代谢产物。斑马鱼经药浴摄入乙酰甲喹后,分别采用乙酸乙酯和乙腈提取组织中的乙酰甲喹及代谢物,过0.22 μm滤膜后经UPLC-Q/TOF-MS分析,通过比较试验组和对照组的色谱图新增色谱峰,确定代谢产物的数量;通过比较乙酰甲喹标准品及代谢产物的精准MS/MS质谱图,确定代谢产物的化学结构,并推测其可能的代谢途径。结果显示,乙酰甲喹在斑马鱼中的代谢产物较少,主要是脱氧代谢物,包括单脱氧代谢物N1-脱氧乙酰甲喹（1-DMEQ）、N4-脱氧乙酰甲喹（4-DMEQ）及双脱氧代谢物N1,N4-双脱氧乙酰甲喹（1,4-BDMEQ）,代谢途径主要为N→O基团还原。残留消除规律研究发现,乙酰甲喹消除较快,4 h降至初始浓度一半以下,并呈现逐渐降低的趋势;其3种代谢产物浓度均呈现先增高后降低的趋势,其中2种单脱氧代谢物在给药后2 h浓度达到最高,8 h降至最高浓度一半以下,双脱氧代谢物在给药后4 h药物浓度达到最高,12 h降至最高浓度一半以下,结果表明,乙酰甲喹在斑马鱼中代谢消除速率较快。上述研究结果可为乙酰甲喹在其他水产食品动物中的代谢研究提供参考,并为水产动物源性食品安全的监控及药代动力学研究提供技术支持。     

乙酰甲喹在斑马鱼中的代谢研究

为了阐明乙酰甲喹在水产动物中的代谢过程,本试验采用超高效液相色谱串联四级杆/飞行时间质谱(UPLC-Q/TOF-MS)技术及代谢软件MetabolynxXS自动分析采集功能,研究其在斑马鱼中的主要代谢产物。斑马鱼经药浴摄入乙酰甲喹后,分别采用乙酸乙酯和乙腈提取组织中的乙酰甲喹及代谢物,过0.22μm滤膜后经UPLC-Q/TOF-MS分析,通过比较试验组和对照组的色谱图新增色谱峰,确定代谢产物的数量;通过比较乙酰甲喹标准品及代谢产物的精准MS/MS质谱图,确定代谢产物的化学结构,并推测其可能的代谢途径。结果显示,乙酰甲喹在斑马鱼中的代谢产物较少,主要是脱氧代谢物,包括单脱氧代谢物N1-脱氧乙酰甲喹(1-DMEQ)、N4-脱氧乙酰甲喹(4-DMEQ)及双脱氧代谢物N1,N4-双脱氧乙酰甲喹(1,4-BDMEQ),代谢途径主要为N→O基团还原。残留消除规律研究发现,乙酰甲喹消除较快,4h降至初始浓度一半以下,并呈现逐渐降低的趋势;其3种代谢产物浓度均呈现先增高后降低的趋势,其中2种单脱氧代谢物在给药后2h浓度达到最高,8h降至最高浓度一半以下,双脱氧代谢物在给药后4h药物浓度达到最高,12h降至最高浓度一半以下,结果表明,乙酰甲喹在斑马鱼中代谢消除速率较快。上述研究结果可为乙酰甲喹在其他水产食品动物中的代谢研究提供参考,并为水产动物源性食品安全的监控及药代动力学研究提供技术支持。     

氯苯胍在鸡组织中的残留消除规律研究

采用超高效液相色谱一串联质谱法（UPLC-MS／MS）研究了氯苯胍在鸡组织中的残留消除规律。白来杭鸡以含氯苯胍500mg／kg饲料饲喂7d,停药后的第0、1、3、5、7天取其肌肉、肝脏、肾脏、皮和脂肪五种组织,采用UPLC-MS／MS法测定其残留状况。结果表明,氯苯胍在肌肉中残留量较小,代谢消除比较快,2d后未检出;而在脂肪、皮和肝脏中残留量较大,代谢消除较慢。     

喹赛多及其两种代谢产物在大鼠体内消除规律研究

通过喹赛多(Cyadox,CYX)在大鼠体内消除规律研究,了解其对食品安全的影响,并为今后的药理学和毒理学研究提供较为详细的数据基础,研究建立了喹赛多及其两种主要代谢产物脱二氧喹赛多(BDCYX)和喹噁啉-2-羧酸(QCA)的提取和HPLC检测方法,并以大鼠作为研究载体,按推荐剂量连续混饲给药7 d后,研究喹赛多及其两种代谢物在血浆、肌肉、肝脏中消除规律;一次性灌胃给药后研究喹赛多及其两种代谢物在排泄物中的消除规律特点。结果表明,CYX和BDCYX在0~24 h和24~48 h时间段的粪便中可大量检出,在血浆、肌肉、肝脏和尿液中未检出;QCA在6 h的肌肉中有少量残留,在肝脏中一直到72 h还有一定量的残留,在血浆和粪便中未发现其存在。本研究结果为今后喹赛多在体内处置研究提供了可直接借鉴的技术手段和理论基础。     

喹乙醇在鲫体内消除规律的研究

报道了鲫单剂量口灌50mg／kg喹乙醇在血浆、肌肉和肝胰脏中消除规律的研究。用乙腈-乙酸乙酯（1／1：V/V）提取组织中的喹乙醇,血浆中喹乙醇用3％的三氯乙酸沉淀蛋白,乙酸乙酯和磷酸盐缓冲液提取喹乙醇代谢物3-甲基喹嗯啉-2-羧酸,超高效液相色谱串联质谱法测定血浆及组织中喹乙醇及其代谢物的浓度。鲫单剂量口灌喹乙醇50mg／kg后,在血浆、肌肉和肝胰脏中的喹乙醇随着时间的延长呈现不同浓度的降解趋势。喹乙醇在血浆中的消除需要4d,在肌肉和肝胰脏中需要7d;喹乙醇的代谢物3-甲基喹嗯啉-2-羧酸在鲫体内消除比较缓慢,肌肉中需要15d,肝胰脏中需要20d。     

地美硝唑在鸽组织中的残留分布及代谢规律研究

为了探索亲鸽哺育乳鸽期间使用地美硝唑后亲鸽和乳鸽体内地美硝唑(DMZ)残留分布及代谢规律，制定适合肉鸽生产的休药期规范。本试验于亲鸽开始哺乳第1天以拌料和灌胃2种方式投喂DMZ,停药后不同时间采集亲鸽、乳鸽组织和鸽乳，采用液相色谱串联质谱法测定DMZ及其代谢物残留量。结果显示，亲鸽采用饲喂和灌胃2种方式投喂DMZ后第1天亲鸽肌肉、肝脏、肾脏和皮脂中DMZ及其代谢物残留量均达到峰值，灌胃方式给药后第1天皮脂中DMZ及其代谢物含量高达6 591.7μg/kg,随后各组织残留量逐渐下降；饲喂方式给药后亲鸽肌肉、肝脏、肾脏和皮脂中药物消除时间分别为16 d、19 d、19 d和19 d,灌胃方式下分别为22 d、22 d、19 d和19 d;乳鸽各组织中药物残留代谢规律与亲鸽基本类似，最高残留量是灌胃后乳鸽皮脂组织，为500.7μg/kg;饲喂方式给药后乳鸽肌肉、肝脏、肾脏和皮脂中药物消除时间分别为16 d、16 d、16 d和13 d,灌胃方式下分别为13 d、19 d、16 d和13 d; 2种给药方式下第10天鸽乳中均无药物检出。结果表明，亲鸽使用DMZ,饲喂方式给药建议亲鸽休药时间为1...     

盐酸土霉素可溶性粉在肉鸡组织中的残留消除研究

在常规养殖环境中,对不同日龄肉鸡分别给予250 mg/L和333 mg/L 50%盐酸土霉素可溶性粉,集中饮水给药,每日1次,连续5 d。停药后4 h,1 d,3 d,5 d,6 d,7 d,8 d,9 d,10 d宰杀肉鸡,采集可食性组织(肌肉、肝脏和鸡胗)样品,肌肉、肝脏和鸡胗组织中的残留量采用UPLC-MS/MS测定。结果表明:土霉素主要残留在肝脏组织中;肌肉、肝脏和鸡胗组织的消除速率相似;停药5 d后,两个剂量组的肌肉、肝脏中土霉素残留量均低于最高残留限量(MRL)。休药期计算软件WTl.4结果显示,上述两种方法给药后,建议休药期分别为7、8 d。     

液相色谱-串联质谱法研究地克珠利在家兔组织中残留及消除规律

[目的]建立一种检测家兔组织中地克珠利含量的液相色谱-串联质谱法（LC—MS／MS）,研究地克珠利在家兔组织中的残留及消除规律。[方法]选择48只家兔（雌雄各半）,在饲料中加入10g／1000kg地克珠利混饲,连续饲喂30d。分别于休药后的第0、1、2、3、4、5、6天,每个时间点随机处死6只家兔（雌雄各半）,取肌肉、肝脏、肾脏组织,DMF,乙腈沉淀法提取并用LC—MS,MS法检测地克珠利残留量。[结果]地克珠利在肾脏中的残留量最高（休药0d为912．6μg／kg）,肝脏次之（休药0d为156．μg／kg）,肌肉最低（休药0d为43．1μg／kg）,在肾脏中的消除速度最快。[结论]该方法适于检测家免组织中的地克珠利含量。休药0d即可满足欧盟要求     

鸡组织中硫酸粘杆菌素残留的微生物学检测方法研究

在国内首次以支气管炎博代特氏菌（Bordetella bronchiseptica）ATCC4617作为检测菌株，用PBS提取鸡肌肉组织，用5％硫酸提取鸡肝脏和肾脏组织中的硫酸粘杆菌素，应用一剂量法设计原理制备肌肉、肝脏和肾脏组织标准曲线。硫酸粘杆菌素检测限肌肉组织为0．15μg／g，肝脏和肾脏组织均为0．20μg／g，低于我国农业部和欧盟规定的粘杆菌素最高残留限量。肌肉、肝脏和肾脏组织标准曲线在0．025—6．4μg／mL范围内线性关系良好，r分别为0．9970、0．9994、0．9990，n=9。以高、低（300mg／L、60mg／L按效价计）两个剂量给鸡饮水给药，肌肉高、低剂量组均未检测出硫酸粘杆菌素残留，肝脏和肾脏低剂量组未检测出硫酸粘杆菌素残留，高剂量组0h有微量残留。     

Distribution and depletion of flubendazole and its metabolites in edible tissues of guinea fowl

1. We measured the distribution and depletion of residues of flubendazole and its major metabolites in breast muscle, thigh muscle and liver of guinea fowls during and after oral administration of the veterinary medicine Flubenol 5% at two doses. 2. The guinea fowls were treated orally with normal feed, medicated at doses of 56 and 86 mg per kg feed for 7 successive days. Afterwards, depletion was observed for 8 d. Just before slaughter, body weights were measured. Thigh muscle, breast muscle and liver of three female and three male birds were sampled. The concentrations of the flubendazole-derived residues were determined by a liquid chromatographic-mass spectrometric method. 3. The highest residue concentrations were obtained for the reduced metabolite. With the therapeutic dose, the maximum mean residue concentrations obtained for this compound in thigh muscle, breast muscle and liver were 312, 288 and 1043 microg/kg, respectively. The values for flubendazole, the parent molecule, were 114, 108 and 108 microg/kg, respectively. The residues of the hydrolysed metabolite were negligible in the sampled muscle tissues. After 24 h of depletion, the sum of the residues of parent and metabolites in muscle tissue still exceeded 50 microg/kg. After 8 d of depletion, flubendazole-derived residues at low concentrations could still be measured in both muscle tissues and liver. Generally, the disposition of residues in breast and thigh muscle was comparable. 4. The European Union has not established a maximum residue limit (MRL) for flubendazole in edible tissues of guinea fowl. In contrast, the existing MRLs for other bird species are expressed as the sum of parent flubendazole and its hydrolysed metabolites. An estimated withdrawal period of three days will assure residue safety in the edible tissues of guinea fowl treated with flubendazole at therapeutic dose. After this withdrawal period following treatment of the guinea fowl, the residues were approximately constant, very low and far below the established safe MRL level for other bird species.     

乙氧酰胺苯甲酯在鸡组织中残留的检测及消除规律的研究

本研究建立了抗球虫药乙氧酰胺苯甲酯在鸡组织中残留的紫外ＨＰＬＣ检测方法。经过对鸡各种组织肌肉、肝、肾、脂肪添加高、中、低三个浓度,每个浓度５个样品,共测定２批,证明笔者建立的提取条件的回收纺在８０％以上,变异系数在１０％以内。最低标准物检测限为１ｎｇ,最低组织检测为２．５ｎｇ／ｇ。     

脱二氧乙酰甲喹及N4-脱一氧乙酰甲喹的合成及结构表征

乙酰甲喹属于喹喔啉-N,N-二氧化物类兽药,有研究认定3-甲基-2-乙酰基-喹喔啉是乙酰甲喹的残留标示物。N4-脱一氧乙酰甲喹则被认为是乙酰甲喹在大鼠、鸡及猪肝微粒体中的主要代谢产物。为给乙酰甲喹代谢产物及代谢机制的深入研究提供理论依据,本研究采用Na2S2O4还原乙酰甲喹,合成3-甲基-2-乙酰基-喹喔啉和N4-脱一氧乙酰甲喹,经质谱、红外及1 H核磁等光谱技术对二者的化学结构进行了确证,为乙酰甲喹代谢产物及代谢机制的深入研究提供了理论依据。     

巨型玫瑰冠鸡H-FABP、A-FABP基因表达的发育性变化及其肌内脂肪含量研究

为探讨心脏型脂肪酸结合蛋白（heart-type fatty acid-binding protein，H-FABP）和脂肪型脂肪酸结合蛋白（adipocyte fatty acid-binding protein，A-FABP）基因与巨型玫瑰冠鸡生长发育及肌内脂肪含量（IMF）的关系，研究H-FABP、A-FABP基因对巨型玫瑰冠鸡IMF含量和腹脂沉积的作用机制，本试验采集了巨型玫瑰冠鸡与良凤花鸡在不同周龄（2、4、6、8、10、12周龄）的腹脂、心肌、胸肌、腿肌组织样品共480份，采用实时荧光定量PCR对巨型玫瑰冠鸡与良凤花鸡不同生长阶段组织中H-FABP、A-FABP基因mRNA表达量进行了检测，并采用索氏浸提法测定了两种鸡12周龄的胸肌与腿肌的IMF含量。结果表明，巨型玫瑰冠鸡与良凤花鸡的H-FABP、A-FABP基因mRNA在腹脂、心肌、胸肌和腿肌组织中均有不同程度的表达，且两种鸡H-FABP基因mRNA在心肌组织中高度表达，在脂肪组织中表达量较低，在胸肌、腿肌组织中中度表达，而A-FABP基因相反，推测H-FABP基因主要在肌肉组织中表达，而A-FABP基因主要在脂肪组织中表达。良凤花鸡生长速度高于巨型玫瑰冠鸡；巨型玫瑰冠鸡的腹脂率均低于同性别的良凤花鸡，但胸肌、腿肌的IMF均高于同性别的良凤花鸡，表明巨型玫瑰冠鸡的肉品质优于良凤花鸡。本试验结果为巨型玫瑰冠鸡H-FABP、A-FABP基因分子选育奠定了基础。     

Tylosin depletion from edible pig tissues

The depletion of tylosin from edible pig tissues was studied following 5 days of intramuscular (i.m.) administration of 10 mg/kg of tylosin to 16 crossbreed pigs. Animals were slaughtered at intervals after treatment and samples of muscle, kidney, liver, skin+fat, and injection site were collected and analysed by high-performance liquid chromatography (HPLC). Seven days after the completion of treatment, the concentration of tylosin in kidney, skin+fat, and at the injection site was higher than the European Union maximal residue limit (MRL) of 100 microg/kg. Tylosin residues in all tissues were below the quantification limit (50 microg/kg) at 10 and 14 days post-treatment.     

乌骨鸡ADFP基因表达的变化及其与脂沉积的相关性

为揭示脂肪分化相关蛋白(adipose differentiation-related protein,ADFP)变化对家禽脂肪性状形成的作用,实验采用SYBRGreenΙ荧光定量法,首次测定了四川山地乌骨鸡1、14、28、42、56、70d和84d胸肌、肝脏、腹脂和皮脂组织中ADFP基因相对表达量。结果表明:ADFP基因表达量在28d,腹脂和肝脏中的表达量显著高于胸肌(P<0.05),且腹脂>肝脏>皮脂>胸肌;在42d,胸肌中的表达量显著高于皮脂(P<0.05),且胸肌>肝脏>腹脂>皮脂;在84d时,胸肌中的表达量显著高于肝脏中的(P<0.05),且胸肌>腹脂>肝脏。腹脂中ADFP基因的相对表达量的变化与活重和冠重性状的变化显著相关(P<0.05),胸肌中ADFP基因的相对表达量发育性的变化与肌内脂肪含量的变化显著相关(P<0.05)。ADFP基因可能在鸡脂肪性状形成的过程中发挥重要的调控作用。     

Dietary exposure assessment of cyadox based on tissue depletion of cyadox and its major metabolites in pigs,chickens, and carp

The tissue kinetics of cyadox, an antibacterial agent used in food animals, and its major metabolites in pigs, chickens, and carp were investigated followed by a complete dietary exposure assessment to evaluate the food safety of cyadox. Cyadox and its major metabolites, bisdeoxycyadox (Cy1), 4‐desoxycyadox (Cy2), N ‐(quinoxaline‐2‐methyl)‐cyanide acetyl hydrazine (Cy4), quinoxaline‐2‐carboxylic acid (Cy6), and 2‐hydromethyl‐3‐hydroxy‐quinoxaline (Cy12), were simultaneously quantitated with a high‐performance liquid chromatography?ultraviolet (HPLC ‐UV ) method. Pigs, chickens, and carp were fed with 150 mg/kg cyadox in feed for consecutive 60, 40, and 30 days, respectively. The residue amount of cyadox and its major metabolites in liver, kidney, muscle, and fat (skin) tissues was determined. Cy2 was below the limit of quantitation even at the withdrawal time of 6 hr, cyadox, Cy4, Cy6, and Cy12 could be detected at 6–24 hr with low level less than 50 μg/kg. By contrast, Cy1 persisted for 3 days in the kidney of pigs and chickens, and in the liver of carp. Based on these residue depletion data and previous toxicology results, the global estimated chronic dietary exposure assessment of cyadox for general population was conducted, indicating a zero withdrawal time (WDT ) may be appropriate for cyadox in food animals when used in feed for prolonged administration. These results provide analytical techniques and safety standards suitable for residue monitoring of cyadox in food animals.     

Residue depletion of decoquinate in chicken tissues after oral administration

Wang, Z.‐Y., Cai, C.‐Y., Chang, X.‐H., Fei, C.‐Z., Qiu, M.‐Q., Jiang, S.‐X., Xue, F.‐Q., Zhang, L.‐F. Residue depletion of decoquinate in chicken tissues after oral administration. J. vet. Pharmacol. Therap.  36 , 116–121. A rapid, sensitive, and reliable high‐performance liquid chromatography tandem mass spectrometry (LC‐MS/MS) method was developed and validated for the analysis of decoquinate in chicken tissues. The compounds were extracted using acetonitrile by liquid–liquid extraction (LLE) and purified with an Oasis^? HLB solid‐phase extraction (SPE) cartridge. Chromatographic separation was performed on an XTerra C₁₈ reversed‐phase column with a mobile phase of water containing 0.1% formic acid and acetonitrile. The analyte was detected by tandem quadrupole mass spectrometry after positive electrospray ionization by multiple reaction monitoring. The detection and quantitation limits were 1 and 2.5 μg/kg, respectively. The recoveries of edible tissues ranged from 85.3% to 104.9%, with relative standard deviations (RSD) lower than 10.4%. The depletion profile of decoquinate was studied in healthy chickens after oral administration of feed containing 27.2 mg/kg decoquinate for 10 consecutive days. The residue concentrations of decoquinate in chicken muscle and liver were detected using the developed method. The highest residue concentrations were attained 0.25 day post‐treatment, and decoquinate residues were still detected 5 days postmedication in the tissues examined. The developed method has been successfully applied to the depletion study of decoquinate in chicken tissues. The recommended withdrawal period with oral administration based on our research is 3 days.