高效液相色谱法检测羊鸡组织中氟苯达唑、噻苯达唑及其代谢物残留量的研究

建立了一种可同时检测羊组织（肌肉、肝脏、肾脏、脂肪）、鸡组织（肌肉、肝脏）中氟苯达唑及代谢物2-氨基氟苯达唑、噻苯达唑及代谢物5-羟基噻苯达唑药物残留的高效液相色谱检测方法。结果表明：氟苯达唑及代谢物2-氨基氟苯达唑、噻苯达唑及代谢物5-羟基噻苯达唑在0.02～1.5μg/mL浓度范围内呈良好的线性关系r,均大于0.999。方法的定量限为20μg/kg。在鸡组织（肌肉、肝脏）中,两种药物及其代谢物在20～800μg/kg的添加浓度范围内,其回收率范围在60%～100%之间;在羊组织（肌肉、肝脏、肾脏、脂肪）中,两种药物及其代谢物在20～200μg/kg的添加浓度范围内,其回收率范围在60%～100%之间;批内变异系数均小于15%、批间变异系数均小于20%。     

牛肉中阿苯达唑及其代谢产物残留的检测

阿苯达唑是一种广谱抗蠕虫药,但是由于用药不当等原因,其在动物可食性组织中的残留可能刺激神经系统及消化系统、触发过敏反应甚至出现肝功能异常。本试验基于5μg/kg的检验限建立牛肉中的阿苯达唑及其代谢产物的检测方法——高效液相色谱法。样品中残留的阿苯达唑及其主要代谢产物阿苯达唑砜和阿苯达唑亚砜,经乙醚提取,乙腈和己烷萃取,由高效液相色谱（配紫外检测器）测定得到阿苯达唑及其亚砜与砜的平均回收率分别为71.8%～88.6%、78.1%～85.6%和83.1%～91.9%,变异系数分别为2.8%～9.2%、0.9%～4.9%和1.4%～4.8%。     

高效液相色谱-荧光法测定牛羊组织中阿苯达唑及其代谢物的残留量

建立了同时测定牛羊组织中阿苯达唑及其代谢物的高效液相色谱-荧光检测方法。试样中待测物经乙酸乙酯提取,酸性氧化铝固相萃取柱和MCX固相萃取柱净化,高效液相色谱-荧光法测定,以外标法定量。药物在5~2000μg/L浓度范围内线性关系良好,相关系数均大于0.999。方法的检出限为0.025 mg/kg,定量限为0.05 mg/kg,在不同组织中添加浓度范围为0.05~10 mg/kg,其平均回收率为60.6%~119.1%,相对标准偏差在1.7%~19.1%之间。方法抗干扰能力强、灵敏度高,适用于牛羊组织中阿苯达唑及其代谢物残留量的测定。     

阿苯达唑亚砜盐酸盐有关物质的HPLC检测

为了建立阿苯达唑亚砜盐酸盐中有关物质的检测方法,本文采用高效液相色谱(HPLC)方法,通过系统适用性、专属性、检测限等试验,确定了阿苯达唑亚砜盐酸盐中有关物质的HPLC检测方法。色谱条件:采用C18色谱柱,流动相为乙腈-蒸馏水(40∶60),检测波长为230nm。结果显示:阿苯达唑亚砜盐酸盐在1.25～40.0μg/mL浓度范围内线性关系良好,检测限为0.032μg/mL。在此条件下阿苯达唑亚砜盐酸盐与其合成原料阿苯达唑、合成中可能产生的阿苯达唑砜、降解产物分离情况良好。     

芬苯达唑微晶体在猪体内的药物动力学试验

本试验以芬苯达唑(FBZ)原料药为对照,考察芬苯达唑微晶体对猪口服给药后FBZ的药动学特征。在0.02~2μg/m L的线性关系良好,相关系数为0.9990。FBZ在猪血浆中的回收率均在70%以上,日内变异系数小于5%,日间变异系数小于10%。芬苯达唑原料组和芬苯达唑微晶体组的主要药物动力学参数为:吸收半衰期(T1/2α)分别为1.65 h和1.07 h,消除半衰期(T1/2β)分别为28.28 h和33.92 h;药时曲线下面积(AUC)分别为29.09μg·h/m L和41.12μg·h/m L。经统计学分析得知,T1/2α、Tp、T1/2β、AUC差异显著(P0.05)。表明芬苯达唑微晶体能够改变芬苯达唑在动物体内的药动学特征,提高芬苯达唑的吸收速度,延长药物在体内的有效作用时间。     

高效液相色谱法测定动物肌肉中的阿苯达唑及其代谢物

建立了动物肌肉组织中阿苯达唑及其代谢物阿苯达唑亚砜、阿苯达唑砜的多残留检测的高效液相色谱法。用乙酸乙酯提取肌肉组织样品中的药物,提取液浓缩后加入酸性乙醇和正己烷分配脱脂,过watersHLB固相萃取柱净化。以水-乙腈-甲酸为流动相,反相高效液相色谱紫外检测法检测。方法的平均回收率为86．4％,平均变异系数为1．96％。3种药物的检测限均为5μg／kg,定量限均为20μg／kg。     

高效液相色谱-串联质谱法检测猪肝中苯并咪唑类药物及其代谢物的残留量

建立了猪肝中阿苯哒唑及其代谢物阿苯哒唑砜、阿苯哒唑亚砜、2-氨基阿苯哒唑砜,噻苯哒唑及其代谢物5-羟基噻苯哒唑,甲苯哒唑及其代谢物氨基甲苯哒唑、5-羟基甲苯哒唑,芬苯哒唑及其代谢物芬苯哒唑砜、等效物奥芬哒唑,氟苯哒唑及其代谢物2-氨基-氟苯哒唑共14种苯并咪唑类药物及其代谢物残留检测的高效液相色谱-串联质谱方法。样品用乙酸乙酯提取,MCX固相萃取柱净化。Waters Xterra C18色谱柱(2.1 mm×150 mm,5μm)分离,流动相A相为0.1%甲酸乙腈溶液;B相为0.1%甲酸水溶液,梯度洗脱,外标法定量。结果表明,14种苯并咪唑类药物及其代谢物标准溶液在10~1 000 ng/mL的浓度范围内呈现良好的线性关系,R2均大于0.99,方法检测限为5μg/kg,定量限为10μg/kg,14种苯并咪唑类药物及其代谢物在10~200μg/kg添加浓度范围内回收率均70%~120%之间。批内、批间相对标准偏差均小于20%。本方法灵敏、准确,满足食品安全检测法规的要求。     

高效液相色谱法测定兽用芬苯达唑粉的含量

建立芬苯达唑粉中芬苯达唑含量的高效液相色谱测定法.采用Agilent 5 HC-C18色谱柱(4.6mm×250mm,5μm),以0.02mol/L磷酸二氢钾溶液:甲醇(20:80)为流动相,流速为1.0ml/min,检测波长为254nm.实验结果表明,芬苯达唑在1.096~43.840μg/ml范围内线性良好(r>0.99998),添加平均回收率为100.9%(n=6),RSD为0.27%.该方法简便、准确、可靠、重现性好,能够满足实际样品的检测要求.     

基于HPLC法测定五氯柳胺阿苯达唑复方混悬液中的有关物质

试验旨在通过高效液相色谱（high performance liquid chromatography,HPLC）法测定五氯柳胺阿苯达唑复方混悬液的有关物质。采用高效液相色谱法,以自身对照法计算五氯柳胺阿苯达唑复方混悬液有关物质的量。选用Thermo Hypersil BDS C18色谱柱（4.6 mm×150 mm,5 μm）;流动相以0.1%磷酸-10 mmol/L磷酸二氢铵水溶液为A相,以甲醇为B相,31：69（V/V）;流速为1 mL/min;进样量为5 μL;柱温30℃;检测波长295 nm。结果显示,在建立的色谱条件下,五氯柳胺阿苯达唑复方混悬液中的主成分峰与杂质峰均能基线分离。3,5,6-三氯水杨酸、2-氨基-4,6-二氯苯酚盐酸盐、五氯柳胺、阿苯达唑在5~100 μg/mL内与峰面积线性关系良好（R² ≥ 0.999）,定量限分别为5 μg/mL及750、500、225 ng/mL,检测限分别为1 μg/mL及500、250、100 ng/mL,杂质3,5,6-三氯水杨酸、2-氨基-4,6-二氯苯酚盐酸盐总平均回收率分别为101.15%、100.35%,总平均回收率RSD均为0.71%（n=9）。经方法学验证,建立的HPLC方法简单、快速、准确,可用于五氯柳胺阿苯达唑复方混悬液中有关物质的测定。     

高效液相色谱法测定牛组织中三氯苯唑及其代谢物残留量的研究

建立了一种可同时检测牛组织中三氯苯唑及其代谢物三氯苯唑酮药物残留的高效液相色谱-紫外检测法。牛组织样品经乙腈提取,异辛烷去脂肪,PPL柱净化。以0.02 mol/L乙酸铵溶液和乙腈溶液为流动相,流速为1.0 m L/min,检测波长296 nm。结果表明,三氯苯唑、三氯苯唑酮在20~1200 ng/m L范围内,峰面积与浓度呈良好的线性关系,R2均大于0.999。方法的最低定量限为50μg/kg。三氯苯唑、三氯苯唑酮在50~600μg/kg的浓度添加水平上,其回收率在70%~100%范围内,批内、批间的相对标准偏差小于20%。     

Characterization of local and peripheral immune system in pregnant and nonpregnant ewes

Modulation of the immune system is known to be important for successful pregnancy but how immune function might differ between the lymph nodes draining the reproductive tract and peripheral lymph nodes is not well understood. Additionally, if immune system changes in response to the presence of an embryo during early pregnancy, and if this response differs in local versus peripheral immune tissue, has not been well characterized. To address these questions, we examined expression of genes important for immune function using NanoString technology in the ampulla and isthmus of the oviduct, endometrium, lymph nodes draining the reproductive tract (lumbo-aortic and medial iliac) as well as a peripheral lymph node (axillary), the spleen, and circulating immune cells from ewes on day 5 of the estrous cycle or pregnancy. Concentrations of estradiol and progesterone in plasma were also determined. Principal component analysis revealed separation of the local from the peripheral lymph nodes (MANOVA P = 3.245e-08, R² = 0.3) as well as separation of tissues from pregnant and nonpregnant animals [lymph nodes (MANOVA P = 2.337e-09, R² = 0.5), reproductive tissues (MANOVA P = 2.417e-14, R² = 0.47)]. Nine genes were differentially (FDR < 0.10) expressed between lymph node types, with clear difference in expression of these genes between the lumbo-aortic and axillary lymph nodes. Expression of these genes in the medial iliac lymph node was not consistently different to either the axillary or the lumbo-aortic lymph node. Expression of IL10RB was increased (FDR < 0.05) by 24% in the reproductive tissue of the pregnant animals compared to nonpregnant animals. Analysis of gene categories revealed that expression of genes of the T-cell receptor pathway in reproductive tract tissues was associated (P < 0.05) with pregnancy status. In conclusion, assessment of gene expression of reproductive and immune tissue provides evidence for a specialization of the local immune system around the reproductive tract potentially important for successful establishment of pregnancy. Additionally, differences in gene expression patterns in reproductive tissue from pregnant and nonpregnant animals could be discerned as early as day 5 of pregnancy. This was found to be associated with expression of genes important for T-cell function and thus highlights the important role of these cells in early pregnancy.     

中华鳖心组织成纤维细胞系的建立及特性

以中华鳖心组织为材料,用含10％胎牛血清的TC199培养基,于25℃条件下,经14个月传代培养60余次,获得一个能稳定生长、以成纤维样细胞为主的细胞系,称之为TSH（Trionyx sinesis heart)细胞系。分别对其原代和传代细胞的染色体数目、不同温度下细胞的生长线及原代和传代细胞的显微形态进行了测定和比较。结果表明,细胞在25℃条件下生长速度最快;原代细胞约有64％的染色体为2n=66,而传代细胞为四倍体的有62％。初步估计的细胞周期为2－3d。传代细胞冻存后,复苏状态良好,能继续增殖传代。     

鸡、兔组织中氯苯胍残留的检测方法研究

研究了用高效液相色谱法(HPLC)检测鸡、兔组织中氯苯胍残留量的方法.用乙酸乙酯提取后离心,上清液减压旋转蒸干,用正己烷溶解残渣后过弗罗里硅土固相萃取柱净化,10%丙酮甲醇溶液洗脱,洗脱液蒸干后用甲醇溶解残渣并定容.HPLC条件:紫外检测器,检测波长353 nm,外标法定量.线性范围在5～500 ng/mL,回归方程为C=139.75A-190.10,r=0.999 9;回收率为70%～110%,批内变异系数小于15%,批间变异系数小于20%;本方法的最低定量限为10 ng/g.     

异生物素及其代谢物在反刍家畜体组织的分泌与排泄机理

胆汁分泌和尿液排泄是异生物素如芳香族和脂环族化合物排出反刍动物有机体的主要途径.渗透过滤是胆汁分泌的主要机制,离子运输对共轭态异生物素的胆汁分泌具有重要作用,其影响因素主要为代谢物极性、分子量和结构.胆汁分泌促进了异生物素在动物体内持续地进行肠肝循环.肾脏中的分泌排泄过程在肾单位中进行.代谢物的尿液排泄包括3个基本过程:肾小球过滤、肾小管分泌和肾小管再吸收.家畜有机体排出异生物素的其他途径有唾液、乳汁、汗水和气体.家畜对某一特定饲料的采食量以及饲料本身的属性直接影响异生物素经尿液的排泄,以此建立合理的关系模型就可以根据尿液中的排泄量测定或预测家畜的采食量.     

The effect of the source and dosage of dietary Cu on redox status in rat tissues

The aim of this experiment was to investigate whether the amount of Cu added to the diet of rats can be reduced without adversely affecting the antioxidant status of tissues and growth, and whether copper nanoparticles can be used for this purpose. For four weeks, four experimental groups of rats were fed diets with two dosages of added Cu (standard—6.5 or 3.25 mg/kg) in two forms (standard—CuCO₃ or copper nanoparticles). Replacing the CuCO₃ supplement with CuNPs resulted in a decreased lung weight and an increased Cu content in brain, kidney and lung, intensification of lipid peroxidation processes, and weakened antioxidant defence in the lungs and kidneys. This treatment also reduced the Cu content in heart, level of lipid oxidation in the liver and testes and improved antioxidant defence in the brain. Reducing the addition of Cu to the diet from 6.5 to 3.25 mg/kg reduced lung weight and increased lipid peroxidation in the liver, heart and lungs, and also weakened antioxidant defence in the lungs and testes. This treatment also weakened the lipid peroxidation process in the spleen, small intestine and brain and strengthened the antioxidant defence of the brain and kidneys. In conclusion, replacing CuCO₃ with CuNPs and reducing the level of Cu in the diet of rats has a particularly unfavourable effect on the respiratory system, causing adverse changes in the lungs. However, these treatments have a clearly positive effect on the redox status of the liver and brain.     

中国与美国、欧盟、日本和CAC猪组织中兽药残留限量标准的对比研究

为便于中国兽药残留标准的改进和相关法规体系的建立提供参考,同时为中国畜产品贸易提供数据参考,保障人们的饮食安全,促进中国畜产品经济的更好发展,本研究首先对中国新国标GB 31650-2019《食品安全国家标准食品中兽药最大残留限量》中规定的猪组织中兽药残留种类和兽药最大残留限量（MRLs）标准进行了简单分析;然后对美国、欧盟、日本和食品法典委员会（CAC）规定的猪组织中兽药残留种类和兽药MRLs标准与中国新国标进行对比,通过对规定兽药总数、中国没有规定而其他标准有规定的兽药种类、其他标准严于中国标准的兽药种类等几项内容的比较,找出差异。结果显示,中国新国标做出了更加详细的补充和修改;从整体上来看,中国规定的猪组织中兽药MRLs标准越来越完善,大部分标准与欧盟、美国、日本和CAC规定标准的差距也越来越小,然而在部分种类兽药MRLs标准的规定方面和这些国家或组织仍有一定的差距。中国需加强与各个发达国家和组织的交流与合作,推进中国的兽药残留标准的发展,加快对动物产品质量安全标准的更新。     

牛多个组织microRNASolexa测序与生物信息学分析

本研究旨在发掘牛更多的microRNA(miRNA)信息及新的miRNA序列,为进一步研究miRNA的生物学功能奠定理论基础.运用高通量测序技术对来自一头西门塔尔公牛和一头荷斯坦母牛的多个组织的小RNA进行混池测序,对测序结果进行了生物信息学分析.随机选取了一条成熟miRNA:bta-miR-2346-5p和两条新预测的miRNAs:novel_miR-48和novel_miR-86,采用茎环RT-PCR进行验证.共鉴定了604条miRNAs,其中429条为牛的已知miRNAs,175条为新预测的miRNAs.通过茎环RT-PCR发现选取的这3条miRNAs在背最长肌、心肌、肝脏、脾脏、肺脏、肾脏、大肠、小肠中均有表达,证明了高通量测序结果的准确性.本试验得到的数据在一定程度上丰富了牛miRNA的数量和种类,为后续牛或其他物种的miRNA相关生物学研究提供了更加详实的信息.     

Distribution and sequence of pyknotic cells in rat fetuses exposed to busulfan

Busulfan, an antineoplastic bifunctional-alkylating agent, is known to induce developmental anomalies. In the present study, we examined the distribution and sequence of pyknotic cells in rat fetal tissues exposed to busulfan. Pregnant rats on gestation day 13 were administered intraperitoneally 30 mg/kg of busulfan, and fetal tissues were examined at 6, 12, 24, 36, 48, 72 and 96 hours after treatment (HAT). Pyknosis of component cells was observed markedly in the brain, moderately in the eyes and spinal cord and mildly in the craniofacial tissue, mandible, limb buds, tail bud, ganglions, alimentary tract, lungs, kidneys, pancreas and liver. In the brain, mitotic inhibition was also detected. Most of the pyknotic cells were considered to be apoptotic cells judging from the results of TUNEL staining and electron microscopic examination. Commonly in the above-mentioned tissues, pyknotic cells began to increase at 24 HAT, peaked at 36 or 48 HAT and disappeared at 96 HAT, which is when the histological picture returned to normal in most tissues except for the brain, spinal cord and eyes. The present study clarified the outline of busulfan-induced apoptosis in rat fetuses.     

高效液相色谱仪检测鸡组织器官喹烯酮残留方法的建立

建立用高效液相色谱法测定鸡组织中喹烯酮浓度的方法,用于监测鸡组织器官中喹烯酮的残留。方法:采用Prodigy 5μPhenyl-3色谱分析柱,Waters TM486可变波长紫外检测器,甲醇-水(70:30)为流动相。组织样品用乙酸乙酯、乙腈、正已烷、氯仿等试剂提取处理纯化后进样,在312 nm波长处检测,流速为1.0 mL／min。标准曲线的线性范围:脂肪、肾脏和肌肉均为0.005-0.080μ／g;肝脏为0.005-0.500μ／g。方法平均回收率:肌肉为(99.23±2.75)％;脂肪为(105.15±8.65)％;肝脏为(93.55±11.16)％;肾脏为(104.10±9.80)％。精密度考察喹烯酮的日内各组织各浓度的RSD(相对标准差)均小于5.1％,日间测得各组织各浓度的RSD均小于8.5％。本方法灵敏度高,重复性好。