高效液相色谱-电喷雾串联质谱法测定动物饲料中的10种磺胺

建立了动物饲料中10种常用磺胺(磺胺嘧啶、磺胺吡啶、磺胺甲基嘧啶、磺胺-5-甲氧嘧啶、磺胺二甲基嘧啶、磺胺甲氧哒嗪、磺胺甲基异唑、磺胺间甲氧嘧啶、磺胺间二甲氧嘧啶(SDM)和磺胺喹啉(SQX))的高效液相色谱(HPLC)-串联质谱检测方法。样品经提取、固相萃取净化、稀释、HPLC分离后进行质谱分析,在多反应监测模式(MRM)下进行特征母-子离子对信号采集。结合保留时间和离子对信息进行定性分析,以共同碎片离子m/z 156进行定量。10种磺胺的定量检测限(S/N=10)为0.5~2.0 μg/kg,在2.0~200 μg/L(SDM和SQX:1.0~100 μg/L)时峰强度与质量浓度的线性关系良好(r>0.9995)。添加水平为1.0 mg/kg时,10种磺胺的平均回收率范围为70%~92%,日内相对标准偏差小于10%,日间相对标准偏差小于15%。结果表明,该法简单、灵敏,特异性强,适用于饲料中多磺胺组分的分析。     

牛奶中12种磺胺类药物残留的高效液相色谱测定方法

建立了高效液相色谱(HPLC)检测牛奶中12种磺胺类药物(磺胺二甲基嘧啶、磺胺间甲氧嘧啶、磺胺甲噁唑、磺胺对甲氧嘧啶、磺胺喹噁啉、磺胺氯哒嗪、磺胺氯吡嗪、磺胺嘧啶、磺胺噻唑、磺胺甲基嘧啶、磺胺甲噻二唑、磺胺地索辛)残留的方法,考察了样品的提取、净化及色谱分析条件。牛奶样经乙酸乙酯提取、固相萃取净化后上机分析,11种磺胺药物标准曲线在10~800μg/L,SM2在5~200μg/L质量浓度范围内相关系数r>0.999,回收率为76%~106%,相对标准偏差小于15.9%。检出限为5~8μg/L,定量下限为14~27μg/L。     

超高效液相色谱法检测化妆品中的12种磺胺抗生素

建立了采用超高效液相色谱(UPLC)-二极管阵列检测器（PDA）测定化妆品中12种常见的磺胺抗生素(磺胺、磺胺间甲氧嘧啶、磺胺醋酰、磺胺甲基异唑、磺胺嘧啶、磺胺二甲异唑、磺胺噻唑、磺胺二甲氧嘧啶、磺胺甲基嘧啶、磺胺喹啉、磺胺二甲嘧啶、磺胺硝苯)的方法。采用Acquity UPLCTM BEHC C18 色谱柱(50 mm×2.1 mm, 1.7 μm),流动相为乙腈/0.1%的甲酸水溶液,梯度洗脱。样品经提取、反萃取后,用UPLC-PDA进行分析检测,结合保留时间和紫外光谱进行定性分析,定量检测波长268 nm。12种磺胺的检出限(S/N=3)均为1 μg/g,定量下限(S/N=10)为2～3 μg/g,在1～25 mg/L(磺胺硝苯为0.5～12.5 mg/L)范围内,峰面积和质量浓度的线性关系良好(r>0.9997)。添加水平为40, 8 μg(磺胺硝苯为20, 4 μg)时,12种磺胺的平均回收率分别为86.8%～98.1%和80.1%～96.9%,相对标准偏差小于10%(n=6)。结果表明该方法简单,分离效果好,速度快,能够满足检测化妆品中12种常见的磺胺抗生素的需要。     

高效液相色谱法同时测定动物源性食品中5种磺胺及3种四环素族抗生素残留

建立了动物源性食品中5种常用磺胺(磺胺甲噁唑、磺胺2甲氧嘧啶、磺胺二甲基嘧啶、磺胺-6甲氧嘧啶、磺胺喹噁啉)及3种四环素族(土霉素、金霉素、四环素)抗生素残留的高效液相色谱同时测定方法.样品经提取,固相萃取净化,浓缩后进行HPLC分离,结合保留时间及二极管阵列检测器(PDA)光谱分析信息定性.添加水平为20、50、100μg/kg时5种磺胺及3种四环素族的平均回收率范围为70%~92%,变异系数小于15%.结果表明,该法简单、快速、灵敏,适用于动物源性食品中多磺胺组份及四环素族组份残留的同时测定.     

超高效液相色谱-串联质谱测定污泥中氯霉素、磺胺类、喹诺酮类、四环素类与大环内酯类抗生素

建立了测定城市污水处理厂污泥中50种抗生素(氯霉素、磺胺、喹诺酮、四环素和大环内酯类)的超高效液相色谱-串联质谱(UPLC-MS/MS)分析方法。污泥样品经乙腈-水溶液超声萃取,高速冷冻离心后,提取液经过HLB柱富集和NH2柱净化;ACQUITY UPLC BEH C18柱用作色谱分离,以0.1%甲酸水溶液-甲醇(ESI+)和乙腈-水(ESI-)为流动相进行梯度洗脱;多反应监测(MRM)模式进行检测;目标药物使用基质外标法定量。50种目标药物的线性范围为0.50～400μg/L,相关系数均大于0.99。方法定量下限为0.10～5.00μg/kg,3个加标水平下的回收率为40%～111%,相对标准偏差为2.9%～27.1%。应用此方法对北京某污水处理厂活性污泥和剩余污泥进行检测,共检出14种药物,包括磺胺嘧啶、磺胺吡啶、磺胺甲唑、吡哌酸、氧氟沙星、依诺沙星、培氟沙星、诺氟沙星、环丙沙星、恩诺沙星、洛美沙星、斯帕沙星、土霉素和阿奇霉素,其浓度为1.2～442.5μg/kg。该方法简便快速、灵敏可靠,适用于污泥中多种抗生素药物残留的同时测定。     

高效液相色谱-串联质谱法对水环境中微量磺胺、大环内酯类抗生素、甲氧苄胺嘧啶与氯霉素的检测

应用固相萃取(SPE)及液相色谱-电喷雾串联质谱(LC-ESI MS/MS)技术,建立了水体中微量磺胺、大环内酯类抗生素、甲氧苄胺嘧啶及氯霉素的定量分析方法.水样中的抗生素通过HLB萃取小柱富集后,以C18反相柱为分析柱,甲醇、醋酸铵和0.1%甲酸混合溶液为流动相,采用LC-MS/MS进行定量分析.选择电喷雾正电离?ESI+),多反应监测模式(MRM),内标法定量.海水和城市污水中抗生素的方法定量下限分别为1.1 ～34.7 ng/L和2.5 ～57.2 ng/L,平均回收率分别为78% ～98%、67% ～111%,相对标准偏差不高于8.8%.通过该方法初步分析了海水及污水中抗生素的含量,检出多种磺胺和大环内酯类抗生素.     

饲料中八种磺胺药物的高效液相色谱测定

结合液、液分配和固相萃取,建立了同时分析食用动物饲料中磺胺嘧啶、磺胺甲基嘧啶、磺胺二甲嘧啶、磺胺甲氧哒嗪、磺胺甲基异唑、磺胺间甲氧嘧啶、磺胺间二甲氧嘧啶和磺胺喹啉8种磺胺类药物的高效液相色谱方法。比较和优化了不同的提取方法和净化参数,方法对饲料中磺胺的检出限为0.20×10-6(w),八种磺胺添加水平为1×10-6(w)时,回收率在68%以上,相对标准偏差小于12%。     

基质固相分散-超快速液相色谱法测定牛肉中磺胺类兽药

采用基质固相分散-超快速液相色谱测定了牛肉组织中磺胺噻唑、磺胺甲基嘧啶、磺胺甲基异恶唑和磺胺间二甲氧嘧啶。基质固相分散萃取的最佳条件为:硅藻土为分散剂,6 mL甲醇为洗脱剂,样品与分散剂的质量比为1∶4。方法的检出限为4.39～7.82 ng/g,定量限为14.62～26.08 ng/g,4种磺胺化合物回收率为71.73%～113.13%,测定的相对标准偏差为1.9%～12.7%。     

高效液相色谱-串联质谱法测定蜂蜜中残留的19种喹诺酮类药物

建立了高效液相色谱-电喷雾串联质谱联用测定蜂蜜中恩诺沙星、环丙沙星、诺氟沙星、氧氟沙星、双氟沙星、恶喹酸、氟甲喹、沙拉沙星、司帕沙星、丹诺沙星、氟罗沙星、马波沙星、伊诺沙星、奥比沙星、吡哌酸、培氟沙星、洛美沙星、西诺沙星和萘啶酸等19种喹诺酮类药物残留的方法。比较酸性溶液阳离子固相萃取(PCX柱)、近中性缓冲溶液反相固相萃取（HLB柱）和碱性溶液阴离子固相萃取(PAX柱)3种不同提取净化方法的提取效果,最终选择使用碱性溶液溶解蜂蜜样品,强阴离子固相萃取柱一步富集净化。以甲醇和0.1%甲酸溶液作为流动相,C18作为分析色谱柱,采用梯度洗脱方式进行液相色谱分离,选择离子反应监测模式检测19种喹诺酮类药物,内标方法定量。在1～100 μg/L范围内,19种喹诺酮类药物的线性相关系数均大于0.991。通过实际样品的添加回收试验,方法的定量限（S/N=10）为1.0 μg/kg,3个添加水平的回收率为71%～118%,相对标准偏差为4.2%～6.7%。     

液相微萃取-反萃取在磺胺药物残留测定中的应用

利用液相微萃取-反萃取技术,建立测定动物组织中磺胺药物残留量的新方法.将动物组织中磺胺药物经液相微萃取-反萃取后进行高效液相色谱紫外检测.磺胺、磺胺嘧啶、磺胺二甲嘧啶、磺胺喹口恶啉4种磺胺类药物的线性范围均在0.05～10.0 μg/mL,磺胺甲口恶唑的线性范围在0.5～10.0 μg/mL之间,方法的平均回收率不低于80.2%,RSD＜6.8%,检出限为0.1 mg/kg.     

Determination of fluoroquinolone antibiotics in surface waters from Mondego River by high performance liquid chromatography using a monolithic column

A novel LC-fluorescence detection method based on the use of a monolithic column for the determination of norfloxacin, ciprofloxacin, and enrofloxacin antibiotic residues in environmental waters was developed. Fluoroquinolones (FQs) were isocratically eluted using a mobile phase consisting of 0.025 M phosphoric acid solution at pH 3.0 with tetrabutylammonium and methanol (960:40, v/v) through a Chromolith Performance RP-18e column (100x4.6 mm) at a flow rate of 2.5 mL/min and detected at excitation and emission wavelengths of 278 and 450 nm, respectively. After acidification and addition of EDTA, water samples were extracted using an Oasis HLB cartridge. Linearity was evaluated in the range of 0.05 to 1 microg/mL and correlation coefficients of 0.9945 for norfloxacin, 0.9974 for ciprofloxacin, and 0.9982 for enrofloxacin were found. The limit of quantification was 25 ng/L for the three FQs. The recovery of FQs spiked into river water samples at 25, 50, and 100 ng/L fortification levels ranged from 76.5 to 91.0% for norfloxacin, 78.5 to 97.2% for ciprofloxacin, and 79.4 to 93.6% for enrofloxacin. This method was successfully applied to the analysis of water samples from the Mondego River, and ciprofloxacin and enrofloxacin residues were detected in eight water samples.     

鸡肉中11种喹诺酮类药物多残留的高效液相色谱检测

建立了用荧光检测器同时测定11种喹诺酮类药物(包括诺氟沙星、培氟沙星、环丙沙星、恩诺沙星、氧氟沙星、达氟沙星、洛美沙星、二氟沙星、沙拉沙星、恶喹酸和氟甲喹)在鸡肉中的多残留的高效液相色谱检测方法。鸡肉样品用10%三氯乙酸-乙腈(体积比为7∶3)提取两次并稀释,随后用反相固相萃取柱净化。采用Hypersil BDS-C18色谱柱分离,以乙腈和水为流动相梯度洗脱,荧光检测器用程序编程检测波长检测。11种喹诺酮类药物标准曲线的线性范围为5～1200 μg/L,相关系数大于0.998。在高、中、低三个添加水平下的回收率为56%～119%,批内相对标准偏差为0.4%～16.1%,批间相对标准偏差为1.4%～23.0%。检出限和定量限分别为1～23 μg/kg和4～40 μg/kg。该方法快速、灵敏,达到了兽药残留检测的要求。     

超高效液相色谱-电喷雾串联质谱法同时分析鸡肉中7种氟喹诺酮类药物残留

建立了一种同时测定鸡肉中7种氟喹诺酮类药物残留的超高效液相色谱-电喷雾串联质谱确证分析方法(UPLC-ESI-MS/MS)。样品经酸化乙腈提取、正己烷脱脂和HLB固相萃取柱净化,采用ACQUITY UPLCTM BEH C18色谱柱(50 mm×2.1 mm,1.7 μm)分离,以0.1%甲酸水溶液和乙腈作为流动相进行梯度洗脱,电喷雾质谱检测,正离子多反应监测模式进行定性和定量分析。7种药物在5～100 μg/kg范围内线性关系良好,相关系数(r2)均大于0.99;以5,25,50 μg/kg3个浓度水平进行添加回收试验,7种药物的平均回收率在79.2%～108.6%之间,相对标准偏差为4.2%～8.9%,方法的检出限(LOD)为0.2～1.4 μg/kg。方法重现性好、灵敏度高、分析时间短、确证能力强,适用于鸡肉中氟喹诺酮类药物多残留的确证检测。     

Multiresidue determination of eleven quinolones in milk by liquid chromatography with fluorescence detection

A liquid chromatographic method with fluorescence detection was developed for simultaneous determination of norfloxacin, ofloxacin, ciprofloxacin, pefloxacin, lomefloxacin, danofloxacin, enrofloxacin, sarafloxacin, difloxacin, oxolinic acid, and flumequine in milk The samples were extracted with 10% trichloroacetic acid/acetonitrile (9 + 1, v/v) and cleaned by Strata-X reversed-phase solid-phase extraction cartridges. The 11 quinolones were separated on a reversed-phase C18 column (Hypersil BDS-C18) with mobile phase gradient elution and detected with fluorescence by means of a wavelength program. The recoveries for milk fortified with the 11 quinolones at 3 levels were 69-88% with acceptable relative standard deviations of <9% (intraday) and <14% (interday). The limits of detection were 23 microg/L for enrofloxacin, and 1-9 microg/L for the other 10 quinolones.     

Direct determination of four fluoroquinolones,enoxacin, norfloxacin,ofloxacin, and ciprofloxacin,in pharmaceuticals and blood serum by HPLC

A rapid, accurate and sensitive method has been developed for the quantitative determination of four fluoroquinolone antimicrobial agents, enoxacin, norfloxacin, ofloxacin and ciprofloxacin, with high in-vitro activity against a wide range of Gram-negative and Gram-positive organisms.A Kromasil 100 C(8) 250 mm x 4 mm, 5 microm analytical column was used with an eluting system consisting of a mixture of CH(3)CN-CH(3)OH-citric acid 0.4 mol L(-1) (7:15:78 %, v/v). Detection was performed with a variable wavelength UV-visible detector at 275 nm resulting in limits of detection: 0.02 ng per 20 microL injection for enoxacin and 0.01 ng for ofloxacin, norfloxacin and ciprofloxacin. Hydrochlorothiazide (HCT) was used as internal standard at a concentration of 2 ng microL(-1). A rectilinear relationship was observed up to 2 ng microL(-1) for enoxacin, 12 ng microL(-1) for ofloxacin, 3 ng microL(-1) for norfloxacin, and 5 ng microL(-1) for ciprofloxacin. Separation was achieved within 10 min. The statistical evaluation of the method was examined by performing intra-day (n=8) and inter-day precision assays (n=8) and was found to be satisfactory with high accuracy and precision. The method was applied to the direct determination of the four fluoroquinolones in human blood serum. Sample pretreatment involved only protein precipitation with acetonitrile. Recovery of analytes in spiked samples was 97+/-6% over the range 0.1-0.5 ng microL(-1).     

高效液相色谱-串联质谱法测定动物组织中的16种喹诺酮类药物残留

建立了动物组织样品中萘啶酸、恶喹酸、氟甲喹、诺氟沙星、依诺沙星、环丙沙星、洛美沙星、丹诺沙星、恩诺沙星、氧氟沙星、沙拉沙星、二氟沙星、麻保沙星、培氟沙星、司帕沙星、奥比沙星等16种喹诺酮类兽药多残留量的高效液相色谱-串联质谱测定方法。用酸性乙腈萃取样品中的16种喹诺酮类药物残留,然后用正己烷脱脂,旋转蒸发浓缩,以Inertsil C8-3色谱柱分离,在正离子模式下以电喷雾电离串联质谱进行测定。在10,50,100 μg/kg 3个加标水平下进行了验证试验,方法的线性范围为10~100 μg/kg,平均回收率为62.4%~102%,相对标准偏差为1.4%~11.9%。该方法简便、快速、准确,各项技术指标满足国内外法规的要求,可用于鸡肉、鸡肝和鱼肉等动物组织样品中喹诺酮类药物多残留的确证检测。     

Ultra performance liquid chromatography tandem mass spectrometry performance evaluation for analysis of antibiotics in natural waters

An ultra performance liquid chromatography electrospray tandem mass spectrometry (UPLC/MS/MS) method was developed and validated for the determination of 17 antibiotics in natural waters in one single extraction and chromatographic procedure. Gradient separation conditions were optimised for 17 compounds belonging to five different antibiotic groups: quinolones (oxolinic acid, nalidixic acid, pipemidic acid, flumequine), fluoroquinolones (enoxacin, ciprofloxacin, norfloxacin, ofloxacin, enrofloxacin, sarafloxacin, danofloxacin, difloxacin, lomefloxacin), sulphonamides (sulphamethoxazole, sulphamethazine), nitro-imidazole (ornidazole) and diaminopyrimidine (trimethoprim). The separation of all compounds, obtained using a 1.7 μm particle size column (100 mm?×?2.1 mm), was achieved within 10 min time. Water samples were adjusted to pH 7 and extracted using Oasis hydrophilic–lipophilic balance (HLB) solid phase extraction cartridges. After elution with methanol and concentration, extracts were injected in a C18 column (Acquity UPLC BEH C18) and detected by tandem mass spectrometry. Average recovery from 100 ng L^?1 fortified samples was higher than 70% for most of the compounds, with relative standard deviations below 20%. Performances of the method (recoveries, detection limit, quantification limit and relative standard deviation) and matrix effects were studied, and results obtained showed that method was suitable for routine analysis of antibiotics in surface water. Samples analysis from Seine River (France) confirmed the interest of antibiotic contamination evaluation in that area.

高效液相色谱-串联质谱法测定葡萄中的吡效隆和赤霉素

采用高效液相色谱-串联质谱法（HPLC-MS/MS）建立了葡萄果实中吡效隆（CPPU）和赤霉素（GA3）两种植物生长调节剂残留量的定性定量分析方法。样品用0.5%甲酸水溶液和乙腈（体积比4:1）混合溶剂提取,经Strata-X固相萃取净化,使用Agilent SB-C18 色谱柱（50mm×2.1mm,1.8μm）,以0.5%甲酸水溶液-乙腈为流动相,流量0.3mL/min,采用电喷雾离子化源（ESI）多反应监测（MRM）模式下HPLC-MS/MS检测。在最佳检测条件下,得到吡效隆和赤霉素在2.0～100.0μg/L检测范围内线性关系良好,相关系数大于0.999;检出限(LOD)分别为0.3μg/L、0.5μg/L,定量限(LOQ)分别为1.0μg/L、2.0μg/L,回收率和精密度均能达到要求,表明该法准确,可用于葡萄中吡效隆和赤霉素残留量测定。     

超高效液相色谱-串联质谱法同时测定土壤中30种抗生素

土壤基质复杂,土壤中残留的抗生素种类繁多,浓度多为痕量水平,高灵敏度的仪器方法、有效的净化和富集方法、多种类抗生素的同时检测是土壤中抗生素检测的重点和难点。该研究建立了固相萃取-超高效液相色谱-串联质谱法同时测定土壤中7类(磺胺类、氟喹诺酮类、四环素类、大环内酯类、β-内酰胺类、酰胺醇类和林可酰胺类)30种抗生素的方法。首先,通过参数优化确定最佳质谱条件,选择BEH-C18色谱柱,以0.1%(v/v)甲酸甲醇溶液-0.1%(v/v)甲酸水溶液为流动相,10%(v/v)甲醇水溶液为进样溶剂。然后,通过提取条件(萃取剂种类及体积)和固相萃取条件(上样液pH、淋洗液有机溶剂比例、洗脱液种类及体积)的优化,确定使用10 mL乙腈和Na₂EDTA-McIlvaine缓冲液的混合溶液(1∶1, v/v)为萃取剂,萃取液pH调节至8.0后,采用HLB小柱进行固相萃取,并以10 mL超纯水淋洗净化,最后用10 mL甲醇-乙腈(1∶1, v/v)洗脱目标分析物。在优化的分析条件下,该方法的定量限为0.043~4.04 μg/kg,目标化合物的标准曲线线性关系良好,相关系数在0.992~1.00的范围内,在20、100、200 μg/kg的添加浓度下,大多数目标化合物的加标回收率范围为44.8%~164%,相对标准偏差为0.700%~14.8%。将该方法用于6个实际土壤样品的分析,结果显示在30种抗生素中,17种抗生素有检出,其中12种抗生素的检出率为100%。环丙沙星和诺氟沙星是土壤样品中含量最高的两种抗生素,它们的含量范围分别是13.7~32.1和15.6~43.6 μg/kg。本研究建立的方法简单、快速、溶剂使用量少,能用于土壤样品中痕量水平的7类30种抗生素的同时测定。     

Development and validation of an HPLC confirmatory method for residue analysis of ten quinolones in tissues of various food-producing animals, according to the European Union Decision 2002/657/EC

The aim of this work was to develop an HPLC method for the simultaneous determination of ten quinolones: enoxacin, ofloxacin, norfloxacin, ciprofloxacin, danofloxacin, enrofloxacin, sarafloxacin, oxolinic acid, nalidixic acid, and flumequine, in various tissues of food-producing animals. Separation was achieved on a PerfectSil Target column (250 mm x 4 mm, ODS-3, 5 microm), by MZ-Analysentechnik (Germany), at room temperature. The mobile phase consisted of 0.1% TFA-CH(3)OH-CH(3)CN and was delivered by a gradient program of 35 min. The detection and quantitation was performed on a photodiode array detector at 275 and 255 nm. Caffeine (7.5 ng/microL) was used as the internal standard (IS). Analytes were isolated from tissue samples by 0.1% methanolic TFA solution. SPE, using LiChrolut RP-18 cartridges, was applied for further purification. The extraction protocol was optimized and the final recoveries varied between 92.0 and 107.4%. The method was fully validated according to Commission Decision 2002/657/EC. Limits of quantitation for the examined quinolones extracted from each tissue were much lower than the respective Maximum Residue Levels, ranging between 30 and 50 microg/kg for bovine tissue, between 30 and 55 microg/kg for ovine tissue, and between 40 and 50 microg/kg for porcine tissue.