建立了高效液相色谱-串联质谱(HPLC-MS/MS)同时检测畜禽粪便中四环素类、磺胺类、喹诺酮类、大环内酯类26种化合物的方法.畜禽粪便冷冻干燥粉碎,HLB固相萃取柱净化,用HPLC-MS/MS进行检测.26种化合物在5～200 ng/mL浓度范围内线性关系良好(r2≥0.99),回收率在70.1％～99.1％之间,日内变异系数范围为1.3％～13.0％,日间变异系数范围为1.6％～14.7％,检测限为2～5 µg/kg,定量限为5～10 µg/kg.本方法操作简便,灵敏度高,适用于猪和鸡粪便中26种化合物的同时测定.

OBJECTIVE A HPLC method for the determination of Determination of Methionine and Vitamin B_1 in Injection of Methionine and Vitamin B_1 was established.METHODS A C_18 column 5μm (4.6×250mm)was used with Acetonitrile Sodium 1-Heptanesulfonate 0.5mol·L~-1 (20 ∶80) as mobile phase. The detection wave length was 225nm.RESULTS The linearity range and average recovery rate for Merhionine were 162.1～648.3μg·mL~-1, r=0.9999, 99.83% and RSD=0.485.while those for Vit B_1 were 16.96～67.84μg·mL~-1, r=0.9998, 99.17% and RSD=1.42.CONCLUSION The method could be used to determine the content of Methionine and Vitamin B_1 in Injection of Methionine and Vitamin B_1 rapidly and acourately.

采用高效液相色谱-四级杆-飞行时间质谱(HPLC-QTOF)筛查并确证1批监督抽检的兽药硫酸黏菌素注射液中的未知添加物,并建立液相色谱-三重四级杆质谱法(HPLC-MS/MS)定量检测20种兽药中15种该类化合物的方法.采用HPLC-QTOF筛查兽药非法添加物时,发现1批硫酸黏菌素注射液中含有未知添加物,与建立的一级谱库中氧氟沙星匹配分数很高,疑似为氧氟沙星.通过二级质谱碎片离子的推导及与对照品二级质谱比对,确证其为氧氟沙星.采用HPLC-MS/MS建立了定量检测20种常见兽药中非法添加氧氟沙星、恩诺沙星、沙拉沙星、达氟沙星、依洛沙星、洛美沙星、麻保沙星、诺氟沙星、环丙沙星、培氟沙星、司帕沙星、二氟沙星、奥比沙星、恶喹酸、氟甲喹共15种喹诺酮类化合物的方法,15种喹诺酮类化合物在各自的线性范围内均呈现良好的线性关系,添加回收率为85.0％～100.0％,相对标准偏差(RSD,n=3)均不高于15.0％.通过检查该样品中的非法添加物,建立了简便、快速、灵敏的定性定量检测方法,为打击当前兽药中非法添加化合物建立技术标准提供参考,为兽药处方外非法添加筛查与确证提供思路.

The method for determination of olaquindox in olaquindox premix by HPLC was developed and validated. Analysis was performed on a C18 column(250 mm×4.6 mm,5 μm)with a mode of 0.02 mol/L monopotassium phosphateaqueous solution(pH value was adjusted to 6.0 by triethylamine)-acetonitrile(92 : 8) at a flow rate of 1.0 mL/min and a column temperature of 30℃. The determination wave length was 262 nm. The results showed that a good linear relationship was accepted in the range of 5~100 μg/mL(R2=1.0000),and the average recovery was 100.2%(n=6)with RSD of 0.2%. This method was convenient,accurate,reliable and was suitable to control the quality of olaquindox premix.

为建立高效液相色谱测定饲料中那西肽A的方法,根据饲料样品多样性的特点,采用乙二胺四乙酸二钠溶液和N,N-二甲基甲酰胺超声提取饲料中的那西肽A.采用C18色谱柱,0.02％磷酸溶液+乙腈(60:40,V/V)为流动相,流速为1.0 mL/min,荧光检测器激发波长为327 nm,发射波长为521 nm.那西肽A在浓度0.02～10μg/mL时线性良好,线性相关系数为0.9999.当添加浓度为0.5～500 mg/kg时,平均回收率为81.6％～95.6％,相对标准偏差(RSD)为1.6％～9.2％.方法的检测限为0.2 mg/kg,定量限为0.5 mg/kg.该方法操作简便、结果准确、稳定性好,应用于实际样品检测,结果满意,表明该方法适用于饲料中那西肽A的测定.

建立了超高效液相色谱-四极杆-飞行时间质谱(UPLC-Q-TOF MS)法快速筛查兽药中72种非法添加化学药物的方法.样品经甲酸-甲醇-水(1:250:250,v/v)溶液提取后,采用Waters Atlantis C18色谱柱(150 mm×3.0 mm,3μm)分离,以5 mmol/L乙酸铵的0.1％甲酸水溶液与5 mmol/L乙酸铵的0.1％甲酸甲醇溶液作为流动相,梯度洗脱,采用正离子模式(ESI+)分析.通过与药物标准品数据库比对,实现了72种药物同时筛查确证.根据实际测定结果和兽药非法添加使用情况,固体样品和液体样品的检出限分别为10 mg/kg和10 mg/L.固体样品在10、20和50 mg/kg(液体样品在10、20和50 mg/L)水平的添加回收率为70.3％～114.8％,相对标准偏差为2.0％～11.5％.203份实际样品中共有18份筛查出非法添加化学药物.本方法分析效率高,结果准确可靠,适用于兽药制剂中非法添加化学药物的筛查.

本研究建立了HPLC-DAD法测定盐酸左旋咪唑注射液中左旋咪唑含量的方法.采用Agilent Eclipse XDB-C18(250 mm×4.6 mm,5μm)色谱柱,流动相为0.05mol/L磷酸二氢钾溶液-乙腈(80:20,v/v),流速为1.0mL/min,进样量为10μL,柱温为30℃,用二极管阵列检测器在190～400nm扫描,记录212nm波长处的色谱图.左旋咪唑在1～500μg/mL浓度范围内线性良好,相关系数为0.99999.在2.5、5.0和10.0mg/g三个添加浓度下,加样回收率为99.1％～100.5％,RSD≤1.5％,检测限为2.5mg/g.本方法简便快捷,准确度和灵敏度高,可用于左旋咪唑注射液的质量控制.

建立并验证了高效液相色谱法测定盐酸二氟沙星粉中二氟沙星含量的方法.采用十八烷基硅烷键合硅胶色谱柱(4.6 mm×250 mm,5μm),以磷酸溶液(取磷酸3.0 mL,加水至1000 mL,用三乙胺调节pH至3.0±0.1,加乙腈50 mL)-甲醇(78:22,V/V)为流动相,流速为1.0 mL/min,柱温为30℃,波长273 nm.试验结果表明二氟沙星在2～100μg/mL范围内线性良好,R2=0.9999,平均回收率为98.1％(n=9),RSD为1.0％.本文开发的方法与农业部公告1960号收载的紫外分光光度法相比具有更好的专属性,同时简便、快速、准确,适用于产品的质量控制.

定性和定量分析一批兽药硫酸黏菌素可溶性粉中的未知添加物.照《中国兽药典》2010年版一部对该批检品采用微生物检定法进行含量测定时,发现该样品的抑菌圈为虚圈,薄层色谱鉴别该样品,未显示与标准品溶液一致的主斑点,怀疑该样品中有处方外非法添加物.采用超高效液相色谱-四级杆-飞行时间质谱(UHP LC-Q/TOF MS)对该样品进行筛查,发现疑似添加物,并使用液相色谱-二极管阵列检测(HPLC-DAD)法进行了双重确证和含量测定.该样品中非法添加物确证为磺胺氯达嗪和甲氧苄啶,添加量分别为58.5 mg/g和13.4 mg/g.

A qualitative and quantitative analysis of unknown additives in a batch of veterinary soluble sulfachlorpyrazine sodium powder had been implemented. The content of soluble sulfachlorpyrazine sodium powder was determined by potentiometric titration. The result showed that the titration was aberrant. According to the standard method, one of the two identifications was unqualified. It revealed that some unknown compounds may be doped in the sample. Then the screening of the sample through ultra high performance liquid chromatography coupled with quadrupole time-of-flight tandem mass pectrometry (UHPLC-Q/TOF MS) showed that the suspicious additives were found in the sample. Furthermore, double confirmation and content determination were accomplished by high performance liquid chromatography-diode array detection (HPLC-DAD). Mequindox and sulfadimidine as the illegal additives were detected at 40.3 and 16.4 mg/g respectively. By summarizing the above progress of deduction, validation and content determination, a good pattern was developed for screening illegal additives in veterinary drugs which provided a reference for the inspection of illegal adulterations in veterinary drugs.

To establish a method for determination of dioxopromethazine illegally added in florfenicol injection. In the course of content determination of 1 batch florfenicol injection with the method of HPLC according to the Chinese Veterinary Pharmacopoeia 2010, a suspicious compound peak was found. The suspicious compound was found by quadrupole time-of-flight tandem mass spectrometry (UHPLC-Q/TOF MS),then confirmation and content determination were completed by high performance liquid chromatography-diode array detector ( HPLC-DAD) . The suspicious compound in this sample was identified as dioxopromethethazine. The results showed that there was a good linear correlation between the peak areas and the concentrations of dioxopromethazine at the range of 1~100 μg/mL( r=1.0000) . The average recovery was 99.5% with RSD of 0.3% by HPLC. The method is convenient, reliable and accurate. It was suitable for detection of dioxopromethazine illegally adulterated in florfenicol injection.

The detection limit,sensitivity,multiple cross reactivity,accuracy and precision of one-step ELISA test kit were estimated,which could be used for the detections of clenbuterol,salbutamol,terbutaline,mabuterol,brombuterol,tulobuterol and cimbuterol simultaneously in swine urine.The results showed that this ELISA kit were suitable for the determinations of 7 kinds of β-agonists residues in swine urine.

The method for content determination of lincomycin in lincomycin hydrochlorde injection by HPLC was developed and validated.Analysis was performed on a C18 column (4.6 mm×250 mm,5 μm) with a mobile phase of 0.02 mol/L diammonium phosphate aqueous solution-acetonitrile (75 ∶ 25,V/V) at a flow rate of 1.0 mL/minand a column temperature of 30 ℃.The determination wave length was 214 nm.The results showed that a good linear relationship was accepted in the range of 0.1 ～4.0 mg/mL (R2=1.0000),and the average recovery was 98.7％ (n =9)with RSD of 0.8％.In contrast with method in Chinese veterinary pharmacopoeia 2015,the nobile phase was optimized by reducing salinity while the peak shape and number of theoretical plates of lincomycin were improved.This method was convenient,accurate,reliable and was suitable to control the quality of lincomycin hydrochlorde injection.

To identify the unknown adulterated compound in one batch of olaquindox premix in the common course,an unknown compound was observed with characteristic UV spectra.The compound was deduced with the information provided by HPLC-DAD.Results showed that the unknown adulterated compound in this sample was identified as quinocetone.To establish a HPLC-DAD method for the determination of quinocetone in olaquindox premix,the chromatographic column was Agilent Eclipse XDB-C18(250 mm ×4.6 mm,5 μm),the mobile phase consisted of a mixture of a solution of methanol and water(60∶40).The flow rate was 1.0 mL/min.The scanning wavelength was from 210 to 400 nm.Quinocetone in 1~100 μg/mL concentration range of linear is good(r=1.0000).The average recoveries was 99.4%~99.8% (n=5)with RSD between 0.8% and 1.0%.

A method for determination of clorprenaline residues in cattle hairs was developed using high performance liquid chromatography coupled with tandem mass spectrometry (HPLC-MS/MS).After hydrolyzed by 1 mol/L hydrochloric acid aqueous solution,purified by MCX olid phase extraction cartridge,the sample was analyzed by HPLC-MS/MS.The calibration curve of clorprenaline was linear in the range of 0.2～800 ng/mL (r≥0.999) and the recoveries of different quantities of clorprenaline in cattle hairs were from 95.9％ to 99.1％ with RSD from 1.1％ to 12.1％.The limit of determination and limit of quantitative of clorprenaline were 0.25 μg/kg and 1.0 μg/kg,respectively.The method was applied for researching the metabolism of clorprenaline in the cattle hairs.The results indicated that the residues of clorprenaline were found in cattle hairs from the fifth day on which the cattle were fed with the drugs to the twenty-first day on which the cattle were not fed.Clorprenaline was easy to accumulate in the cattle hairs and difficult to be metabolized.This study provides the reference and basis for supervision of edible animal feeding process.

建立反向高效液相色谱法检测白龙散中龙胆苦苷和小檗碱含量的方法.试验采用十八烷基硅烷键合硅胶色谱柱(4.6 mm×250 mm,5μm),以0.5％三乙胺水溶液(用85％磷酸调pH值至3.0)-甲醇(60∶40,v∶v)为流动相,流速为1.0 mL/min,柱温为30 ℃;进样量为10 μL;检测波长为270 nm;龙胆苦苷和小檗碱的含量测定范围分别为4.09 ～ 102.2 μg/mL(R =0.9997),3.93 ～98.26 μg/mL(R=1.0000);加样平均回收率分别为98.63％,97.78％;RSD分别为0.96％,1.23％;该方法方便、准确、可靠,适用于测定白龙散中龙胆苦苷和小檗碱的含量.

To perfect the quality control method of rhubarb powder.Gallic acid was identified by thin layer chromatography (TLC)and determined by high performance liquid chromatography (HPLC).A chromatographic column of Agilent Eclipse XDB -C18(250 mm ×4.6 mm,5 μm)was used,with the mobile phase of methanol 0.05% phosphoric acid solution (5 ∶95)at the flow rate of 1.0 mL/min.The detection wavelength was at 271 nm.The calibration curves were linear in ranges of 1.064 ~106.4 μg/mL(r =1.0000)for gallic acid;the average recoveries (n =6)was 97.84%(RSD =0.61%).The method is accurate and sensitive for the quality control of rhubarb powder.

An analytical HPLC -MS/MS method was established for determination of diclazuril in feeds.The samples were extracted with acetonitrile,then defatted with n -hexane.The treated samples were analyzed by HPLC-MS/MS after centrifuged.The analysis was performed with Multi Reaction Monitor(MRM)in negative ion mode.The results indicated good lineartites in the concentration range from 0.5 to 100 ng/mL for diclazuril, the correlation coefficient was over 0.999.The limit of detection(LOD)and the limit of quantitation(LOQ)were 0.1 and 0.5 μg/kg,respectively.While the spiked contents of diclazuril standard at 0.5,1,5 μg/kg,the recoveries and relative standard deviations(RSD)were in range of 79.2% ~94.5% and 1.7% ~14.3%, respectively.With the advantages of good anti -interference ability,rapidness and sensitivity,the method adapted to the determination of diclazuril in feeds.

To establish a RP -HPLC method for determination of andrographolide and ehydroandrographolide in Siwei Chuanxinlian San,analysis was performed on a C18 column (250 mm ×4.6 mm,5 μm)with a gradient elution mode of methanol -0.2% phosphoric acid at a flow rate of 1.0 mL/min and a column temperature of 30 ℃.The determination wave length of andrographolide was 226 nm while dehydroandrographolide was 252 nm. The linear range was 2.32 ~37.14 μg /mL(r =1.0000)and 2.00 ~31.94μg /mL (r =1.0000),and the average recovery was 99.12% and 98.84% with RSD of 0.60% and 0.62%,respectively.This method was convenient,accurate and reliable for the determination of andrographolide anddehydroandrographolide in Siwei Chuanxinlian San.

In order to improve the risk assessment of chemicals residue in animal products,the basic theory of risk assessment and progress of risk assessment of chemicals residue in animal products were both summarized,and then the distinctiveness of risk assessment of chemicals residue was analyzed.