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参照《中国兽药典》2000版收载的细菌内毒素检查法，验证了硫酸卡那霉素注射液的细菌内毒素检查法。通过干扰试验证明，硫酸卡那霉素注射液对鲎试剂的凝集反应无干扰作用，用灵敏度为0．25EU／mL的鲎试剂检查细菌内毒素的方法可行、有效。可以用细菌内毒素检查法替代家兔热原检查法来检测硫酸卡那霉素注射液的热原。 相似文献
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浅谈细菌内毒素检查法中的计算方法 总被引：3，自引：1，他引：2
鉴于我国细菌内毒素检查的应用情况和兽药药品检测的实际情况,结合工作实践,对细菌内毒素检查法中的细菌内毒素限值（Ｌ）、最大有效稀释倍数（ＭＶＤ）、最小有效浓度（ＭＶＣ）、鲎试剂灵敏度复核（λｃ）及供试品干扰试验的判断（Ｅｓ、Ｅｔ）的计算方法进行了多次试验,Ｅｔ在０．５Ｅｓ～２．０Ｅｓ（包括０．５Ｅｓ和２．０Ｅｓ）时,供试品在该浓度下不干扰试验。相反存在增强或抑制作用。 相似文献
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免疫亲和色谱-气相色谱/质谱法(IAC-GC/MS)检测猪肝中的沙丁胺醇与克伦特罗 总被引：6，自引：0，他引：6
制备出抗沙丁胺醇的抗血清，提取、纯化IgG制备免疫亲和色谱柱，针对沙丁胺醇与克伦特罗的动态柱容量分别为400ng／mL基质和416ng／mL基质。猪肝样品匀浆后用稀盐酸提取，经免疫亲和色谱柱纯化，再用GC／MS检测，即为IAC-GC／MS法。对沙丁胺醇的检测限为0．5ng／g，定量限为2ng／g，对克伦特罗的检测限为0．8ng／g，定量限为2．5ng／g。空白组织按照1、5、10、20ng／g的浓度添加药物，沙丁胺醇在空白肝中的添加回收率为78．4％～106．9％，克伦特罗的添加回收率为77．1％～102．9％。本研究建立的检测猪肝中沙丁胺醇与克伦特罗的IAC-GC／MS法乃国内首次报道。 相似文献
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本文建立了泰地罗新原料药细菌内毒素检查方法。根据《中国兽药典》2015年版一部附录1143细菌内毒素检查法(凝胶法)的要求[1],通过干扰试验确定样品最大不干扰浓度,并进行方法学验证。结果显示泰地罗新质量浓度小于0.4mg/mL时,不会干扰内毒素和鲎试剂的反应;泰地罗新内毒素应小于1.25EU/mg。结论:可以采用此法对泰地罗新原料药进行细菌内毒素检查。 相似文献
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建立了同时测定饲料中21种磺胺类药物和13种喹诺酮类药物的超高效液相色谱-串联质谱方法。以2％氨水乙腈和0．1 mol／L 氢氧化钠溶液提取样品中的待测物,HLB固相萃取柱净化,液相色谱-串联质谱法测定,外标法定量。药物在1～50 ng／mL浓度范围内线性关系良好,相关系数均大于0．99。方法的检出限为0．05 mg／kg,定量限为0．10 mg／kg,在不同饲料中添加浓度范围为0．1～100 mg／kg,平均回收率为60．6％～118．6％,相对标准偏差在1．9％～15．2％之间。本方法抗干扰能力强、分析速度快、灵敏度高,适用于饲料中磺胺类和喹诺酮类药物的测定。 相似文献
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对油剂普鲁卡因青霉素注射液细菌内毒素检查方法进行了研究,在样品中加入适量的无菌、无内毒素的０．１ｍｏｌ／Ｌ氢氧化钠％吐温－８０溶液;并用０．１ｍｏｌ／Ｌ盐酸调节样品液的ＰＨ值,解决了样品的溶解方法,。用稀释法排除样品对细菌内毒素检查的干扰。实验结果表明,选用鲎试剂灵敏度为０．５ＥＵ／ｍｌ将样品稀释 １２５００ｕ／ｍｌ的溶液后可消除其干扰作用,因此,用细菌内毒素检查法作为该药品的热原物质检查是可行持 相似文献
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猪肝和猪尿中沙丁胺醇和克伦特罗残留的酶联免疫吸附检测法研究 总被引：18，自引：1，他引：18
采用混合酸酐法将沙丁胺醇与牛血清白蛋白偶联，并免疫家兔制备抗沙丁胺醇抗血清。以间接ELISA法测定血清效价，测得抗血清最佳工作浓度为1：6400，该抗沙丁胺醇抗血清对克伦特罗有110％的交叉反应性。建立了能够同时检测SAL和CL的间接竞争ELISA方法，该方法对沙丁胺醇的检测范围为0．48ng／mL～8．0μg／mL，对克伦特罗的检测范围为0．43ng／mL～7．3μg／mL，对沙丁胺醇的检测灵敏度为o．48ng／mL，对克伦特罗的检测灵敏度为0．43ng／mL。在猪肝和猪尿中添加1～50ng／g(ng／mL)的沙丁胺醇，添加回收率为72．2％～84．8％，变异系数为4．2％～28．3％，在猪肝脏和猪尿中添加1～50ng／g(ng／mL)的克伦特罗，添加回收率为84．3％～103．4％，变异系数为O．3％～35．5％。 相似文献
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为调查我国当前主流饲喂方式下奶牛静脉血内毒素浓度的范围及内毒素浓度与产奶量的相关性,本试验首先验证了应用鲎试剂法定量奶牛静脉血中内毒素浓度的可行性,在此基础上,分别检测了安徽牧场 １（６０头,饲粮精粗比 ４５∶５５）、江苏牧场（１５头,按产奶量高低精粗比分别为 ２０∶８０、４５∶５５、５０∶５０）和安徽牧场 ２（１４头,饲粮精粗比 ７０∶３０）共 ８９头奶牛的尾静脉血中内毒素浓度,并进行了内毒素浓度与产奶量的相关性分析。结果表明：鲎试剂法可有效检测奶牛静脉血中内毒素浓度,３个牧场奶牛静脉血中内毒素浓度高低不等,在 ０～１．５０ＥＵ／ｍＬ范围内。奶牛静脉血中内毒素浓度与产奶量之间的相关性皆不显著（Ｐ＞０．０５）,但安徽牧场 ２的相关系数（０．１９５２）大于安徽牧场 １（０．０２０５）和江苏牧场（０．０３７６）。结果提示,鲎试剂法可用于定量检测奶牛血中内毒素浓度,在我国当前的饲喂方式下,奶牛血液中普遍含有内毒素,静脉血内毒素浓度与产奶量无显著相关性。 相似文献
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本文旨在对严重影响奶业健康发展的奶牛蹄叶炎的病因进行探究。在辉山奶牛场阜新市彰武县三场经产奶牛中,挑选临床健康奶牛10头作为对照组;患蹄叶炎奶牛10头作为试验组。采集两组奶牛血液,分别用牛内毒素酶联免疫试剂盒和荧光分光光度法测定血浆中内毒素和组胺的含量。结果显示,试验组血浆内毒素及组胺含量分别比对照组高0.65ng/mL、21.8ng/mL,差异均极显著(P<0.01)。说明患病牛血浆内毒素和组胺含量升高是诱发奶牛蹄叶炎的重要因素。 相似文献
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The purpose of this experiment was to explore the influence of the sample loading volume,flow rate and protein concentration of protein on the removal efficiency of endotoxin and protein loss in porcine visfatin recombinant lipoprotein solution at 4 ℃.In this study,commercial and modified polymyxin B was used as the medium to specifically remove endotoxin from the protein solution.At 4 ℃,different loading volume (6,8,10,12 mL),flow rate (0.125-0.75 mL/min) and protein concentration (0.9,1.2,1.3,1.7 mg/mL) were used for endotoxin removal,the content of endotoxin was determined by a quantitative method using endotoxin reagents,and the BCA protein concentration assay kit was used to determine the protein concentration.The results showed that at 4 ℃ endotoxin removal was the best when protein loading volume was 12 mL and flow rate was 0.125-0.25 mL/min,the removal rate was 99.5%,the residual amount of endotoxin (1 EU/mL) was within the biosafety range,and the protein recovery rate was as high as 90% to 95%.The experimental results showed that the endotoxin was successfully removed from the solution of porcine lipin recombinant protein,which provided references for the related functional research and clinical application of the protein. 相似文献
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本试验旨在探索4 ℃条件下蛋白的上样体积、流速和蛋白浓度对猪内脂素重组蛋白溶液中内毒素的去除效果和蛋白损失量的影响。该研究采用商品化、修饰过的多黏菌素B为介质，特异性地去除蛋白溶液中的内毒素。在4 ℃条件下，选用不同的上样体积（6、8、10、12 mL）、流速（0.125~0.75 mL/min）和蛋白浓度（0.9、1.2、1.3、1.7 mg/mL）进行内毒素的去除，使用内毒素检测试剂盒定量测定内毒素含量，使用BCA蛋白浓度测定试剂盒进行蛋白浓度的测定。试验结果表明：在4 ℃条件下，蛋白上样体积12 mL、流速0.125~0.25 mL/min时，内毒素的去除效果最佳，去除率高达99.5%，内毒素的残余量（1 EU/mL）在生物安全范围以内，并且蛋白的回收率高达90~95%。试验结果证明成功去除猪内脂素重组蛋白溶液中的内毒素，为该蛋白的相关功能研究和临床应用提供参考。 相似文献
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According to the present study the limulus amebocyte lysate test (LAL) seems to be a convenient test to detect endotoxin in milk from udder quarters with and without inflammation. The correlation between endotoxin concentration and the results from the bacteriological investigation of 79 milk samples was good (Table I). Determination of endotoxin in 20 milk samples from cases of acute clinical mastitis with high cell count and a negative bacteriological culture showed that all but one had an endotoxin concentration of greater than 1.0 ng/ml milk (Table II). By using a micromethod of the LAL it is possible to detect cases of mastitis caused by gram-negative bacteria about one hour after the sample has reached the laboratory. In a preliminary field study milk from 13 cases of acute clinical mastitis were tested by a modified limulus amebocyte lysate (LAL) test ("cowshed test"). A 100% correlation to bacteriological findings was observed (Table IV). By using the LAL test to detect mastitis cases caused by gram-negative bacteria great economic advantages and less risk for resistance problems can be achieved by using proper antibiotics. This is the fact in Sweden where the frequency of acute clinical mastitis caused by streptococci (100% of strains sensitive for penicillin) and Staphylococcus aureus (about 90% of strains sensitive for penicillin) is high (70-80%) and about 20% are caused by gram-negative bacteria, mostly E. coli. 相似文献
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This experiment used carbodiimide get egg albumin synthesis of artificial antigens coupling neomycin,through square test to determine the best antigen packaged concentration and antibody dilution ratio,mass concentration logarithm of neomycin as abscissa,inhibition rate of antibody as the ordinate drawing standard curve,established ELISA fast detection techniques of neomycin in edible animal for import and export.Methods of recovery and specific test had been done in further research.Results showed that the test dilution ratio of optimum synthetic antigen was 1:200,the best dilution multiple of antibodies was 1:2 000, half inhibitory concentration(IC50) was 6.99 ng/mL;The minimum detection limit of the method was 40 ng/mL,and cross reaction rate of gentamicin, kanamycin, streptomycin,tobramycin,amikacin and dihydrostreptomycin were less than 0.1%,the average of recovery rate was 83.77%.The range of standarding curve in 1.5~40 ng/mL was linear,the correlation coefficient was 0.987,the equation of standarding curve was y=-37.66x+94.592;Compared with HPLC method,the operation was simple and fast.The test results showed that the enzyme-linked immunosorbent method of detecting neomycin was rapid,efficient and specific. 相似文献
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应用碳二亚胺法合成人工抗原得到卵清蛋白偶联新霉素,通过方阵试验确定最佳抗原包被浓度与抗体稀释倍数,以新霉素质量浓度对数为横坐标、抗体抑制率为纵坐标绘制标准曲线,建立了进出口食用动物中新霉素ELISA快速检测技术,并对建立的方法进行了回收率和特异性试验。结果表明,建立的检测方法包被人工合成抗原的最适稀释度为1：200,抗体的最佳稀释度为1：2 000,半数抑制浓度（IC50）为6.99 ng/mL;该方法的最低检测限达40 ng/mL,与庆大霉素、卡那霉素、链霉素、托普霉素、阿米卡星、双氢链霉素的交叉反应率均小于0.1%,平均加标回收率83.77%。标准曲线在1.5~40 ng/mL范围内是线性的,相关系数为0.987,标准曲线方程为y=-37.66x+94.592;与常规液相检测方法相比操作简单、快速。本研究建立了一种快速、高效、特异的新霉素ELISA检测方法。 相似文献
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This study was aimed to prepare excellent fluorescence immunochromatographic strip for clenbuterol (CLB) which could achieve the purpose of rapid and quantitative detection and laid a foundation for its industrialization.The strip was prepared through single factor experiments using latex particles as a fluorescent label.The preparation process was optimized with CLB concentration,drying time,mixing reaction time,reaction temperature and chromatography time.And the properties of this strip including detection limit,specificity,clinical accuracy and added recovery,degree of precision and stability were investigated.Optimum modification conditions were shown as following:CLB concentration was 2.0 mg/mL,drying time was 5 d,mixing reaction time was 1 min,reaction temperature was 25 ℃,chromatography time was 15 min.The detection limit was up to 0.35 μg/L and the cross rate between CLB and other eight similar drugs were less than 0.8% which demonstrated the good specificity.The added recoveries of each batch were between 80% and 120% which met the requirements.The coefficient of variation (CV) under different concentrations of CLB was less than 5% which met the precision standard.Besides,the stability of the experiment also showed that the test strip had good stability. 相似文献
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本研究旨在制备性能优异的克伦特罗(clenbuterol,CLB)荧光免疫层析快速定量检测试纸条,为实现其产业化应用奠定基础。采用荧光胶乳微粒为标记物,以抗原包被浓度、膜干燥时间、混匀反应时间、反应温度及层析时间为单因素优化制备了CLB荧光免疫层析试纸条,并对该试纸条的检测限、特异性、临床准确度及添加回收率、精密度和稳定性进行了评价。最佳制备条件:抗原包被浓度2.0 mg/mL,膜干燥时间5 d,混匀反应时间1 min,反应温度25 ℃,层析时间15 min。评价结果显示,试纸条检测限可达0.35 μg/L;CLB与其他8种同类药物交叉反应率均小于0.8%,特异性良好;各批添加回收率在80%~120%之间,准确度达到要求;不同CLB浓度下变异系数(CV)均小于5%,符合精密度标准;此外,稳定性试验验证了该试纸条稳定性良好。 相似文献
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Bacterial coldwater disease, caused by Flavobacterium psychrophilum, has lead to the loss of significant numbers of hatchery-reared salmonids. The bacteria can be spread from parent to progeny within contaminated sperm and ovarian fluid and can enter the egg during fertilization. The addition of antibiotics to diluents and water-hardening solutions could prevent the spread of the disease. In separate trials, a mixture of 0.197 mg/mL penicillin plus 0.313 mg/mL streptomycin was added to both a 0.5% sodium chloride fertilization diluent and hatchery well water during hardening. Tests showed that the addition of the antibiotics to the diluent and during up to 60 min of water hardening had no effect on the eye-up, hatch and deformity rates of Rainbow Trout Oncorhynchus mykiss eggs compared with the nonantibiotic-treated controls. Also, significant reductions in the prevalence of F. psychrophilum on the surface and inside eggs were observed when compared with controls. These results indicate that the addition of penicillin and streptomycin to diluents and during water hardening can prevent the vertical transmission of bacterial coldwater disease.
Received May 1, 2014; accepted July 10, 2014 相似文献