杀菌剂中抑霉唑含量的离子对高效液相色谱分析

选用庚烷磺酸钠作离子对试剂 ,拟定了杀菌剂中抑霉唑及其硫酸盐的反相离子对HPLC测定方法。讨论了离子对的形成过程 ,pH值对离子对形成的影响 ,推导了流动相pH值计算公式 ,估算了流动相的pH值范围 ,确定了色谱柱为EclipseXDB C815 0mm× 4 .6mm ,流动相为甲醇 :含 12mmol/L庚烷磺酸钠的 2 5mmol/L磷酸盐缓冲溶液 (pH≈ 2 .8) =6 2∶38(V/V) ,测定波长为 2 2 5nm的分析条件。方法的检出限为 7.1× 10 -4μg ;线性范围为 8.0× 10 -3 ～ 4 .0 μg ,实验结果线性关系良好 ;相关系数r =0 .9998。方法用于杀菌剂样中抑霉唑及其硫酸盐的测定 ,回收率 99.7% ,结果满意     

固相萃取-液相色谱-串联质谱法检测小鼠肌肉组织中矮壮素

建立了快速检测小鼠肌肉组织中矮壮素的固相萃取-液相色谱-串联质谱(SPE-LC-MS/MS)方法。小鼠肌肉样品经乙腈提取,弱阳离子交换(WCX)固相萃取柱净化,3 mL甲酸-甲醇(1∶99,V/V)重力洗脱。采用亲水作用色谱柱(HILIC),含10 mmol/L乙酸铵、0.1%甲酸水溶液-乙腈(40∶60,V/V)为流动相,以电喷雾正离子(ESI+)、多反应监测模式(MRM)进行矮壮素的定性分析,采用基质标准曲线外标法进行定量分析。结果表明:矮壮素的线性范围为5.0～500.0μg/L,线性相关系数为0.9991。在10.0,100.0和200.0μg/kg添加浓度下的回收率为73.2%～82.3%;相对标准偏差小于9.3%;方法定量限为10.0μg/kg,能够满足小鼠肌肉组织中痕量矮壮素检测的要求。     

雪莲果中甲霜灵、异菌脲和除虫脲残留量的高效液相色谱测定方法

采用高效液相色谱法测定雪莲果中甲霜灵、异菌脲和除虫脲农药残留量.通过乙腈提取,固相萃取(SPE)净化,Hypersil Cold(4.6 mm i.d.×250 mm,5 μm)柱,V(乙腈)∶V(20 mM NH4Ac)=70∶30为流动相,柱温30℃,流速1.0mL/min进行分离,进样20 μL,2487双波长紫外检测器进行检测.添加回收率为78.4%～92.1%;RSD分别为3.6%～6.3%;甲霜灵,异菌脲和除虫脲检出限分别为0.05,0.04,0.02 μg/mL.     

高效液相色谱-串联质谱测定海水中氯霉素残留量

应用高效液相色谱-串联质谱法测定海水中的氯霉素残留量, 流动相为V(甲醇)∶V(水)＝80∶20. 采用负电喷雾离子源, 在多反应性监测模式下采集信号, 定性离子对为321/121、 321/152、 321/176, 定量离子对为321/152. 本方法的线性范围是0.1～100 μg/L, 相关系数为0.9993, 仪器检出限为0.01 μg/L, 回收率在81.7%～121.0%之间. 该方法适于海洋环境监测.     

亲水作用色谱法测定表阿霉素及其杂质

建立了一种以亲水作用色谱分离测定表阿霉素的新方法。采用硅胶色谱柱及高极性有机溶剂水相缓冲溶液流动相。对流动相的pH、缓冲溶液的浓度及流速进行了优化,确定了以乙腈甲酸钠缓冲溶液(pH2.9)(90∶10,V/V)作为流动相的最佳条件。对优化的分离条件进行系统适应性实验,结果表明表阿霉素与有关杂质之间的分离度和拖尾因子均达到药典要求。该法具有良好的线性(相关系数0.9971～0.9991)和重复性(峰面积RSD<1.0%),方法简便实用,用于实际样品分析,结果满意。     

亲水作用色谱法测定胡芦巴中的胡芦巴碱

建立了亲水作用色谱法(HILIC)测定胡芦巴药材中胡芦巴碱含量的方法。采用Waters Atlantis HILIC Silica色谱柱(150 mm×2.1 mm, 3 μm),以乙腈-乙酸铵溶液(pH 4.4)(体积比为70:30)为流动相,流速0.4 mL/min,检测波长265 nm。胡芦巴碱的线性范围为2.50～100 mg/L (r=0.9996);两个加标水平的平均加样回收率为102%,相对标准偏差(RSD)分别为4.17%和2.28%(n=3)。结果表明所建方法分离效果好、快速简易,可以弥补中国药典中离子对色谱法(IPLC)平衡时间过长的缺陷,适用于胡芦巴药材中强极性胡芦巴碱的测定,为胡芦巴的质量控制提供了有效的方法。     

高效液相色谱柱后衍生测定鸡组织中甲基盐霉素残留量

建立了鸡组织中甲基盐霉素的高效液相色谱柱后衍生化分析方法.样品经异辛烷提取,离心后上层有机相过硅胶固相萃取小柱,洗脱液浓缩后用V(甲醇)∶ V(水)=90∶ 10混合液溶解.采用Inertsil ODS-3 C18柱,以V(甲醇): V(乙酸)∶ V(水)=94∶ 3∶ 3为流动相,香草醛为衍生剂进行高效液相色谱柱后衍生分析,520 nm检测,外标法定量.方法检出限为6 μg/kg; 定量限为20 μg/kg; 添加浓度在20～1800 μg/kg范围内,平均添加回收率为76.4%～93.1%; 批内相对标准偏差(RSD)在2.6%～8.9%之间; 批间相对标准偏差(RSD)在4.7%～9.7%之间.样品浓度在0.07～10.0 mg/L范围内与峰面积呈良好的线性关系,r>0.9993.     

固相萃取-高效液相色谱法测定乳及乳制品中土霉素、四环素和金霉素残留

建立了固相萃取-高效液相色谱同时测定乳及乳制品中土霉素、四环素和金霉素的方法。样品经pH 4.0的Na2EDTA-mcllvaine缓冲溶液漩涡超声提取后,采用Oasis HLB固相萃取(SPE)小柱净化,洗脱液用N2吹至近干后用流动相溶解并定容,过PTFE滤膜后在Shiseido MG C18色谱柱上,以V(乙腈):V(10 mmol/L草酸)=27∶73的溶液为流动相进行分离,检测波长为360 nm。在最优的色谱条件下,各目标化合物的加标回收率为83.0%~109.2%,相对标准偏差在2.8%~6.1%之间。3种抗生素含量在0.10~20.0 mg/L范围内线性关系良好,相关系数≥0.999,乳及乳制品中土霉素、四环素和金霉素的检出限分别为10,10和20μg/kg。     

反相离子对液相色谱法同时测定11种氟喹诺酮类药物的研究

建立了高效液相色谱-荧光法同时测定11种氟喹诺酮类药物的分析方法.主要研究了流动相、流动相配比及流动相的pH对氟喹诺酮分离的影响.确定了液相色谱分析最佳条件.分离条件为:Xbridge Shield RP C18柱,以V(0.10%三氟乙酸)∶V(乙腈)∶V(甲醇)=89∶4∶7为流动相;检测波长为λex=280 nm和λem＝450 nm.方法检出限为:诺氟沙星、环丙沙星、培氟沙星和恩诺沙星0.007μg/mL,单诺沙星0.002 μg/mL,沙拉沙星和氧氟沙星为0.04 μg/mL,二氟沙星和奥比沙星为0.02 μg/mL,依诺沙星、麻保沙星为0.4 μg/mL,各组分回收率在97%～100.2%,相对标准偏差为0.2%～2.9%.     

液相色谱-电喷雾串联质谱法测定冬青芽中的脱落酸

建立了测定植物中痕量脱落酸 (ABA) 的液相色谱-电喷雾串联质谱联用分析方法. 植物提取液先采用固相萃取 (SPE) 富集脱落酸并消除基体干扰, 然后以C18柱为固定相, V(甲醇)∶V(2 g/L甲酸水溶液)＝50∶50为流动相分离脱落酸. 电喷雾(ESI)串联质谱采用负离子模式. 选择反应监测 (SRM) 模式用于脱落酸定量, 选择的离子对是263→153, 219. 脱落酸的线性范围为0.005～10 μg/mL, 检测限 (S/N=3) 为0.003 μg/mL, 加标0.02 μg和0.05 μg的回收率分别为98.3% 和103.5%. 该方法用于冬青芽中脱落酸的分析, 结果表明所建立的SPE-LC-MS/MS方法对于分析植物中的痕量脱落酸是有效的.     

Ion chromatographic analysis of tetracyclines using polymeric column and acidic eluent

High-performance ion chromatography (HPIC) is first successfully used to analyze tetracycline antibiotics (TCs) in this work. The TCs are well separated on a solvent compatible polymeric cation-exchange column within 12 min. Isocratic elution with acetonitrile-hydrochloride is very advantageous for routine analysis. HPIC may be seen as a specific variant of the more common high-performance liquid chromatography (HPLC) for water-soluble and polar pharmaceuticals with low hydrophobicity. The detection limits (signal-to-noise ratio=3:1) of oxytetracycline (OTC), tetracycline (TC), chlortetracycline (CTC), doxycycline (DC) are 10, 10, 20 and 20 microg l(-1), respectively. Samples are prepared by vortex mixing with an ethylenediaminetetraacetic acid disodium salt (Na2EDTA)-McIlvaine buffer (pH 4.0) solution and the mixture filtrates through a molecular weight cut-off filter. The method has been successfully applied to monitor the OTC removal rate through every reactor in the process of OTC manufacturing wastewater treatment by bio-chemical technology. It is also applicable to determine the TCs residues in milk and milk powder with satisfying results.     

养殖废水中四环素HPLC分析方法的研究

为了评价养殖废水中四环素类抗生素残留的状况,建立了养猪场废水中四种常见四环素类抗生素(土霉素、四环素、金霉素、强力霉素)的高效液相色谱分析方法.水样中四环素类药物经HLB固相萃取小柱吸附,甲醇乙酸乙酯溶液洗脱后用高效液相色谱紫外检测器测定.在0.05～10.00 μg/mL.范围内,四种抗生素的峰面积与质量浓度的线性关...     

Hydrophilic interaction chromatography separation mechanisms of tetracyclines on amino-bonded silica column

Effects of mobile-phase variations on the chromatographic separation on amino-bonded silica column in hydrophilic interaction chromatography (HILIC) were investigated for four zwitterionic tetracyclines (TCs): oxytetracycline, doxycycline, chlortetracycline, and tetracycline. A mixed-mode retention mechanism composed of partitioning, adsorption, and ion exchange interactions was proposed for the amino HILIC retention process. Buffer type and pH significantly influenced the retention of TCs, but showed similar separation selectivity for the tested analytes. Experiments varying buffer salt concentration and pH demonstrated the presence of ion exchange interactions in TCs retention. The type and concentration of organic modifier also affected the retention and selectivity of the analytes, providing direct evidence supporting the Alpert retention model for HILIC. The retention time of the analytes increased in the following order of organic modifiers: tetrahydrofuran < methanol < isopropanol < acetonitrile. The linear relationships of logk' versus %water (v/v) curve and logk' versus logarithm of %water (v/v) in the mobile phase indicated that TCs separation on the amino phase was controlled by partitioning and adsorption. The developed method was successfully utilized in the detection of TCs in both river water and wastewater samples using solid-phase extraction (SPE) for sample cleanup.     

四环素类抗生素-钨酸钠-乙基紫体系的共振瑞利散射光谱及其分析应用

在pH为9.0的Clark-Lubs缓冲溶液中, 强力霉素、土霉素、四环素和金霉素等四环素类抗生素与钨酸钠反应形成1∶1的阴离子螯合物, 它仅能引起吸收光谱的变化, 不能引起共振瑞利散射(RRS)的增强, 但是当该螯合物进一步与乙基紫反应形成三元离子缔合物时, RRS显著增强并产生新的RRS光谱, 它们具有相似的光谱特征, 最大RRS波长均位于328 nm处. 4种抗生素的线性范围和检出限分别为0.047～4.8 μg•mL^－1和14.1 ng•mL^－1(强力霉素); 0.078～5.0 μg•mL^－1和23.5 ng•mL^－1(土霉素); 0.081～5.7 μg•mL^－1和24.4 ng•mL^－1(四环素); 0.122～7.7 μg•mL^－1和36.6 ng•mL^－1(金霉素). 考察了三元离子缔合配合物的组成, 讨论了配合物的结构和反应机理, 并发展了一种高灵敏、简便快速测定四环素类抗生素的新方法.     

Determination of tetracyclines in food samples by molecularly imprinted monolithic column coupling with high performance liquid chromatography

A novel solid phase extraction (SPE) method for determination of tetracyclines (TCs) in milk and honey samples by molecularly imprinted monolithic column was developed. Using tetracycline (TC) as the template, methacrylic acid (MAA) as the functional monomer, ethylene glycol dimethacrylate (EGDMA) as the cross-linker, methanol as the solvent, cyclohexanol and dodecanol as the mixed porogenic solvents, a TC imprinted monolithic column was prepared by in situ molecular imprinting technique for the first time, and the optimal synthesis conditions and the selectivity of TC imprinted monolithic column were investigated. The interfering substances in food samples and TCs can be separated successfully on imprinted column. Molecularly imprinted solid phase extraction (MISPE) coupling with C18 column was used to determinate the TCs in milk and honey. The recoveries of this method for six tetracyclines antibiotics such as tetracycline (TC), oxytetracycline (OTC), minocycline (MINO), chlortetracycline (CTC), metacycline (MTC) and doxycycline (DTC) were investigated, and high recoveries of 73.3-90.6% from milk samples and 62.6-82.3% from honey samples were obtained. A method for determination of TCs at low concentration level in milk and honey samples was successfully developed by using the monolithic column as the precolumn for solid phase extraction of six TCs compounds.     

Improvement of chemical analysis of antibiotics. X. Determination of eight tetracyclines using thin-layer and high-performance liquid chromatography

Analytical methods for eight tetracyclines (TCs) were established using silica gel high-performance thin-layer chromatography (HPTLC), reversed-phase thin-layer chromatography (RP-TLC) and high-performance liquid chromatography (HPLC). Good separations of eight TCs were obtained using chloroform-methanol-5% disodium ethylenediaminetetraacetate solution (65:20:5) (lower layer) and methanol acetonitrile 0.5 M oxalic acid solution (1:1:4) (pH 3.0) on silica gel HPTLC and C8 TLC plates, respectively. A combination of HPTLC and RP-TLC made possible the identification of the eight TCs. Each calibration graph was linear between 0.1 and 1.0 microgram using UV densitometry except for rolitetracycline. For detection reagents, the diazonium salts including Fast Violet B gave variously coloured spots with the eight TCs and good sensitivities were obtained except with minocycline. In HPLC, the simultaneous analysis of the eight TCs on a C8 column was possible using methanol-acetonitrile-0.01 M oxalic acid solution (1:1.5:7) adjusted to pH 3.0 as the mobile phase. A linear relationship was obtained between 1.0 and 10 ng using the usual sample preparation except for rolitetracycline. The direct determination of rolitetracycline was possible using tetrahydrofuran, dimethyl sulphoxide and the mobile phase as solvents for preparation of the sample. For the determination of residual rolitetracycline, it was effective to measure the amount of rolitetracycline as tetracycline by HPLC, HPTLC and RP-TLC after conversion of rolitetracycline to tetracycline by incubating for 5 min in methanol at 50 degrees C.     

反相高效液相法测定四环素类抗生素

 利用高效液相法 ,在C18柱上以甲醇 乙腈 0 0 1mol/L草酸溶液 (pH 2 0 ) (体积比为 11∶2 2∶6 7)为流动相 ,采用 2 6 7nm紫外光进行检测 ,在 2 2min内将 7种四环素类抗生素全部洗脱并达到基线分离。探讨了流动相的pH值、草酸的浓度、流动相中有机相的比例以及检测波长等因素对分离度和灵敏度的影响。采用标准加入法定量 ,对两种实际样品进行了分析。结果表明 :该方法操作简单、灵敏度高、定量准确。     

Determination of tetracycline antibiotics by reversed-phase high-performance liquid chromatography with fluorescence detection.

A highly sensitive method for the determination of tetracycline antibiotics (TCs) using reversed-phase high-performance liquid chromatography with fluorescence detection is presented. This method was based on the use of disodium ethylenediaminetetraacetate (EDTA) and calcium chloride as fluorescence-increasing reagents in the mobile phase. The concentrations of each reagent in the mobile phase greatly influenced the fluorescence intensity of TCs. When the concentration of EDTA and calcium chloride were 25 and 35 mM, respectively, and the pH of the mobile phase was 6.5, the maximum fluorescence intensity was obtained. The column temperature hardly influenced the fluorescence intensity. At 3.75 ng of TCs injected, the precision (relative standard deviation) ranged from 1.12 to 2.20%. In the range 0.075-37.5 ng for tetracycline and oxytetracycline and 0.225-37.5 ng for chlortetracycline, a linear response was observed. The detection limits of this method were 49-190 pg for three different TCs. The proposed method was applied to the determination of one of the TCs in pharmaceuticals by the internal standard method using other TCs as internal standards and was also applied to determination of TCs added to fish tissue.     

高效液相色谱化学发光法检测牛奶中残留四环素类化合物的研究

基于四环素类抗生素药物中的四环素(TC)、土霉素(OTC)、金霉素(CTC)和多西环素(DC)能够强烈增敏通过恒电流电解方法在线电生BrO^－和鲁米诺之间产生的化学发光, 提出了一种高效液相色谱(HPLC)化学发光(CL)法检测4种四环素类抗生素药物的新方法. 以Nucleosil RP-C₁₈ (250 mm×4.6 mm, i.d., 5 μm, pore size, 100 Å)为色谱柱, 0.05 mol• L^－1磷酸二氢钾(pH 2.5)-乙腈(30∶70, V∶V)为流动相, 流速1.2 mL/min, 柱温25 ℃, 同时分离检测四种抗生素的总时间为11 min. 研究并优化了流动相、电生试剂化学发光检测的条件. 四种抗生素的检出限为0.002～0.008 μg•mL^－1 (3σ), 对0.01 μg•mL^－1的四种抗生素测定的相对标准偏差为2.0%～3.6% (n＝11). 该方法已成功应用于牛奶中残留四环素类抗生素含量的分析.     

Simultaneous extraction and analysis of 11 tetracycline and sulfonamide antibiotics in influent and effluent domestic wastewater by solid-phase extraction and liquid chromatography-electrospray ionization tandem mass spectrometry

Wastewater treatment plants (WWTPs) in which antibiotic compounds are not totally eliminated are considered to be point sources of antibiotic contamination in surface and ground waters. Therefore, there is a need for sensitive and reliable analytical methods for measuring these compounds in WWTP water matrices. This paper describes a simultaneous method for the determination of six tetracyclines (TCs) (oxytetracycline (OTC), tetracycline (TC), demeclocycline (DMC), chlortetracycline (CTC), doxycycline (DXC), meclocycline (MCC)) and five sulfonamides (SAs) (sulfathiazole (STZ), sulfamethazine (SMT), sulfachloropyridazine (SCP), sulfamethoxazole (SMX) and sulfadimethoxine (SDM)) using solid-phase extraction followed by liquid chromatography-ion trap tandem mass spectrometry. The average recovery of 11 antibiotics for simultaneous extraction was 83.3+/-12.6 and 89.8+/-11.5% for six TCs, and 95.2+/-11.4 and 97.7+/-10.6% for five SAs in the influent and effluent water, respectively. Matrix effects were found to be significant when measuring TCs but not SAs. The accuracy and day-to-day variation of the method fell within an acceptable range of 15% absolute. Method detection limits in wastewater matrices were between 0.03 and 0.07 microg/L. For the investigated 11 antibiotic compounds TC, DMC, CTC, DXC, SMT, SMX and SDM were found in the influents with a concentration range of 0.05-1.09 microg/L. CTC, DXC and SMX were also detected in the effluents with a concentration range of 0.06-0.21 microg/L. These results were compared with those in WWTP effluents of Canada, Germany and Switzerland.