Determination of uric acid in canine serum and urine by high performance liquid chromatography

A reproducible high performance liquid chromatography (HPLC) method was developed for analysis of uric acid in canine serum and urine. The method consists of precipitating serum proteins with phosphotungstic acid prior to HPLC analysis. Urine is analyzed after dilution with buffer. Chromatography is performed on a reversed-phase C-18 column with UV detection at 292 nm. Sensitivity of the method will allow reproducible measurement of uric acid at concentrations of 0.05 mg/dl in serum and 0.1 mg/dl in urine. The HPLC method has been used to quantify hundreds of canine serum and urine samples. The method is superior to UV absorption or colorimetric methods because its lower limit of detection allows measurement of uric acid at concentrations found in canine serum and urine.     

高效液相色谱法测定鸡饲料中噁喹酸的含量

建立了测定饲料中口恶喹酸含量的高效液相色谱分析方法。采用乙酸乙酯提取饲料中的口恶喹酸。检测条件为:磷酸溶液-乙腈-四氢呋喃(69∶16∶15,V∶V∶V)作为流动相;C18色谱柱;激发波长325 nm;发射波长369 nm。结果表明,浓度在10～500 ng/mL,线性关系良好,r=0.999。在不同浓度添加水平下,三种鸡饲料中口恶喹酸的平均回收率为85.75%～96.77%,日内变异系数为2.24%～9.63%,日间变异系数为2.35%～8.35%,方法检测限为5.0μg/kg,定量限为16.0μg/kg。     

牛、羊奶中莫昔克丁残留高效液相色谱荧光检测方法的建立

建立了适用于牛、羊奶样品中莫昔克丁残留的高效液相色谱荧光检测方法。以乙腈为提取液,C18固相萃取柱净化,并用N-甲基咪唑-乙腈(1+1,V/V)和三氟乙酸酐-乙腈(1+2,V/V)进行衍生化。乙腈+水(90+10,V/V)为流动相,荧光法定量。经测定,该方法对牛、羊奶中莫昔克丁的检测限为2 μg/kg,定量限为5 μg/kg。空白牛、羊奶中标准品莫昔克丁添加量分别为5、20、40和80 μg/kg时,牛奶中莫昔克丁平均回收率范围为75.5%～110%,其批内变异系数小于738%,批间变异系数小于7.87%;羊奶中莫昔克丁平均回收率范围为77.5%～95.6%,其批内变异系数小于5.06%,批间变异系数小于5.92%。该方法准确性强、灵敏度高,适用于牛、羊奶中莫昔克丁的快速检测。     

超高效液相色谱法测定饲料中地昔尼尔

本研究建立了超高效液相色谱测定饲料中地昔尼尔的方法。饲料样品经过乙腈提取,取5mL上清液氮气吹干,用20%甲醇溶液(含0.2%甲酸)溶解,经过Waters Oasis MCX固相萃取小柱净化,然后用Waters Acquity UPLC HSS T3色谱柱(2.1mm×100mmi.d.,1.8μm)分离,以甲醇和水为流动相进行洗脱,外标法定量。结果表明,地昔尼尔质量浓度在0.05～100μg/mL内线性良好,相关系数大于0.9999;在10～200mg/kg的添加水平,平均加标回收率为78.0%～86.3%;相对标准偏差为4.2%～6.1%;方法的检出限低至0.05mg/kg,定量下限低至0.1mg/kg。该方法简单、快速、灵敏,可满足饲料中地昔尼尔的快速筛选检测的要求。     

Identification of metoclopramide metabolites in the urine of cattle by gas chromatography-mass spectrometry and high-performance liquid chromatography-photodiode array detection

The urinary metabolites of metoclopramide (4-amino-5-chloro-N-[2-diethylaminoethyl]-2-methoxybenzamide) were identified in cows. The drug was administered intravenously, voided urine was collected, and individual urine extracts were analysed by gas chromatography-mass spectrometry and high-performance liquid chromatography-photodiode array detection. The parent compound and one major metabolite (4-amino-5-chloro-N-[2-(ethylamino)ethyl]-2-methoxybenzamide) were common to all individuals. In addition to the parent and major metabolite, a second, minor metabolite was identified in two cows as 4-amino-5-chloro-N-[2-(diethylamino)ethyl]-2-hydroxybenzamide. The identity of the minor metabolite was confirmed by comparison with a standard synthesized by a new method. Metabolite identification and characterization in food animal species allows the design of safety and environmental impact studies and relative metabolite ratios between dose treatment groups.Abbreviations ¹H-NMR proton nuclear magnetic resonance - R _T retention time - D₁ dopamine-1 - D₂ dopamine-2 - 5-HT₃ 5-hydroxytryptamine - GC-MS gas chromatography-mass spectrometry - HPLC high-performance liquid chromatography - UV-vis ultraviolet-visible - ID internal diameter - m/z mass/charge ratio     

Effects of phenylbutazone and oxyphenbutazone on basic drug detection in high performance thin layer chromatographic systems

Interference or 'masking' in thin layer chromatography occurs when the presence of one drug on a thin layer plate physically obscures or interferes with the detection of another drug. We investigated the ability of phenylbutazone and oxyphenbutazone to mask or interfere with the detection by high performance thin layer chromatography (HPTLC) of basic drugs used illegally in horse racing. Of fifty-five basic drugs called 'positive' since 1981 by laboratories affiliated with the Association of Official Racing Chemists (AORC), forty did not comigrate with phenylbutazone or oxyphenbutazone and could not, therefore, be masked. When 75 micrograms/ml of oxyphenbutazone was spiked into urine samples, subjected to an extraction procedure for basic drugs, and then run in our routine HPTLC systems, no 'spots' due to oxyphenbutazone appeared. 'Masking' by oxyphenbutazone, therefore, did not and could not occur in our test systems. When phenylbutazone at a concentration of 30 micrograms/ml was spiked into urine samples and run in the routine HPTLC system, phenylbutazone spots were visible under ultraviolet light and after certain specific oversprays were used to visualize basic drugs. These spots, however, did not interfere with routine thin layer testing for basic drugs. It was concluded that phenylbutazone and oxyphenbutazone had no significant ability to interfere with detection of the parent forms of these basic drugs under the conditions described in these experiments.     

动物性食品中卡巴氧标示残留物检测方法--高效液相色谱法

农业部于2003年1月22日发布第236号公告,发布12种动物性食品中兽药残留检测方法。方法之1　范围本标准规定了动物性食品中卡巴氧的标示残留物喹啉口恶2羧酸(ＱＣＡ)残留量检测的制样和高效液相色谱测定方法。本标准适用于猪的肌肉、肝脏和肾脏组织中卡巴氧标示残留物残留量检测。2　规范性引用文件下列文件中的条款通过本标准的引用而成为本标准的条款。凡是注日期的引用文件,其随后所有的修改单(不包括勘误的内容)或修订版均不适用于本标准,然而,鼓励根据本标准达成协议的各方研究是否可使用这些文件的最新版本。凡是不注日期的引用文件,其…     

高效液相色谱法测定喹乙醇含量

喹乙醇具有促进动物生长、提高饲料转化率及广谱抗菌作用。相对其他兽药 ,添加成本较低 ,大部分饲料厂 ,特别是中小饲料厂多用其作为畜禽配合饲料的药物添加剂。由于喹乙醇具有中等至明显的蓄积毒性 ,饲料中添加量过大或混合不均匀、饲喂时间过长等均能引起鸡、鸭中毒。我国目前正加紧实施饲料安全工程 ,加大对违规使用饲料药物添加剂行为的打击力度。开展饲料及饲料添加剂中喹乙醇含量检测工作非常重要 ,参考有关资料 ,根据喹乙醇的理化特性 ,笔者摸索出了一套饲料及饲料添加剂中喹乙醇含量的测定方法 ,方法简便、灵敏、准确 ,现介绍如下。…     

高效液相法测定伊维菌素粉的含量

伊维菌素是从阿佛曼链霉素菌发酵产物分离的独特物质经化学修饰而制成。20世纪80年代中期的国外研究证实,伊维菌素对动物体内线虫和体表寄生虫均有显著驱杀效果。1994年美国默沙东药厂生产伊维菌素,年销售额达4.95万美元,列居当年世界动物保健品市场的10大产品榜首。由伊维菌素加一定辅料配制而成的伊维菌素粉,具有广谱、低毒、安全、适口性好、无刺激性及用法简便等特点。经临床试验证明,对驱杀畜禽肠道线虫,体表螨、虱、蜱效果良好,明显降低畜禽死亡率,增重迅速,提高饲料转化率和畜禽产品质量。但目前只收载过伊维菌素注射液含量的测定方法,伊维菌素粉剂尚无收载和报道。本法采     

高效液相色谱-二极管阵列检测法测定乳酸环丙沙星可溶性粉中非法添加甲硝唑和替硝唑的研究

为确证乳酸环丙沙星可溶性粉中的非法添加物,采用高效液相色谱-二极管阵列检测器(HPLC-DAD)的谱库匹配,推断出疑似添加物为甲硝唑,从而建立检测乳酸环丙沙星可溶性粉中违规添加甲硝唑和替硝唑的高效液相色谱法。采用Agilent Eclipse XDB-C18色谱柱(250 mm×4.6 mm,5μm),流动相为甲醇-水(20∶80,V∶V),流速为1.0 mL/min,柱温30℃,用二极管阵列检测器进行扫描,扫描范围为190～400 nm,检测波长为320 nm。该色谱条件下甲硝唑和替硝唑与其他物质峰分离良好。甲硝唑和替硝唑在0.5～100μg/mL浓度范围内线性良好,在2.5 mg/g、5.0及10.0 mg/g 3个添加浓度下,加样回收率为99.5%～100.2%,RSD小于1.5%,检测限为2.5 mg/g。本方法简便快捷、准确度和灵敏度高,能满足非法添加物检测的要求。     

高效液相色谱法测定饲料中拉沙洛西钠

拉沙洛西钠是由拉沙洛链霉菌产生的一种芳香聚醚类离子载体型抗生素 ,主要作为畜禽专用抗生素用于治疗和预防肉鸡和绵羊的球虫病 ,并能提高和改善牛的饲料利用率 ,对动物有促进生长和提高饲料报酬的作用。但饲料中添加剂量过大会对动物产生毒性作用 ,并会造成兽药残留等问题 ,因此美国和欧洲等国家均规定了其在饲料中的允许限量 ,我国农业部规定了饲料中拉沙洛西钠的允许限量为 75～ 1 2 5mg/kg。世界各国均先后进行了饲料中拉沙洛西钠测定方法以及动物血、尿和组织中拉沙洛西钠残留量的检测方法研究。目前应用最多的有荧光分光光度法、薄层…     

高效液相色谱法检测饲料中的着色剂

实验建立了同时分离检测饲料中叶黄素、斑蝥黄、β-胡萝卜素3种着色剂的高效液相色谱法。样品用乙腈-丙酮提取,以甲醇-二氯甲烷为流动相在1.2 mL/min的流速下进行梯度洗脱。该方法在6 min内实现了3种着色剂的基线分离,叶黄素、斑蝥黄的线性范围为0.05～200μg/mL,β-胡萝卜素的线性范围为0.5～200μg/mL,线性相关系数≥0.9991。叶黄素、斑蝥黄、β-胡萝卜素的检测限分别为0.025、0.020、0.25μg/mL;样品平均回收率为80.1%～105.0%,相对标准偏差<5.00%。     

高效液相色谱法对饲料中赭曲霉毒素的测定

本实验研究了高效液相色谱法测定饲料原料和配合饲料中赭曲霉毒素的方法。样品经乙腈水(84∶16,V∶V)提取,提取液通过Mycosep229柱净化、浓缩,C18色谱柱分离,荧光检测器检测,外标法定量对添加2个浓度赭曲霉毒素的玉米样品进行加标回收实验,平均回收率为99.12%和103.09%,变异系数分别为0.281%、1.214%,最低检测限为0.20ng/kg,方法简便易行,准确度、精密度高。     

高效液相色谱法测定饲料中环丙氨嗪

本试验研究建立了用NH2柱和97%乙腈水溶液做流动相洗脱测定饲料中灭蝇胺———环丙氨嗪含量的高效液相色谱法,饲料均质后经20%氨水乙腈溶液重复提取2次、浓缩处理后上机检测,环丙氨嗪标准曲线在0.05～20.0μg/mL线性关系良好,相关系数(R)为0.99995;在饲料中环丙氨嗪含量为3.0～8.0mg/kg时,回收率为87.23%～95.36%,变异系数为3.04%～3.46%。检测限为0.05mg/kg。     

高效液相色谱法测定饲料中的利血平

本实验建立了饲料中利血平的测定方法。采用高效液相色谱仪,乙酸铵缓冲液+乙腈=60+40作为流动相,C18色谱柱,激发波长:280nm,发射波长:360nm。结果表明:在0.01～10μg/mL浓度范围,线性关系良好,r=0.9999。试样中利血平的检出限为0.05mg/kg。本法操作简便,测定结果准确。     

高效液相色谱法测定饲料中替米考星含量

为建立饲料中替米考星含量的高效液相色谱检测分析方法,实验样品经水-乙腈-磷酸二丁胺溶液-四氢呋喃（805:115:25:55,V:V:V:V）提取后,振荡10 min,超声10 min,4000 r/min离心5 min,取上清液过0.45 μm有机微孔滤膜后供高效液相色谱仪分析测定。结果表明：替米考星在10 ~ 500 μg/mL质量浓度范围内线性关系良好,相关系数大于0.999,方法检出限为5 mg/kg,定量限为10 mg/kg,平均回收率为92.0% ~ 101.2%,相对标准偏差为1.02% ~ 5.17%。该方法操作简单,重复性好,准确率高,且上机8 min即可完成替米考星的检测,适用于饲料中替米考星含量的测定。[关键词] 饲料|替米考星|高效液相色谱法