高效液相色谱法测定水产品中四环素类抗生素残留

建立了一种高效液相色谱法测定水产品中土霉素、四环素、去甲基金霉素、金霉素、脱氧土霉素的分析方法。样品用5.0%高氯酸溶液提取,上清液用OasisHLB固相萃取柱净化,用紫外检测器于355nm测定。土霉素、四环素、去甲基金霉素检测限为0.01mg/kg,金霉素、脱氧土霉素检出限为0.02mg/kg。5种药物的回收率在74.8%~89.3%之间,相对标准偏差为3.95%~9.95%。方法适用于水产品中四环素类抗生素残留的检测。     

高效液相色谱同时测定饲料中三种抗生素

建立了反相高效液相色谱法同时检测饲料中土霉素、四环素、金霉素的方法.色谱柱为AichromBond-AQ C18柱,150×4.6mm,粒度5μm;流动相:0.01mol/L磷酸二氢钠溶液(pH2.5)+乙氰,采用梯度洗脱,乙氰浓度在0～13min内由13%上升到40%;柱温35℃;流速为1.0mL/min;进样量101μL;检测波长375nm.方法检出限:土霉素和四环素0.25μg/mL、金霉素0.50μg/mL,回收率91.70%～101.19%,RSD<1.39%(n=7).     

分子印迹固相微萃取-高效液相色谱法测定水和牛奶中三种四环素类药物

以土霉素为模板分子制备了分子印迹固相微萃取涂层,建立了选择性萃取、高效液相色谱法同时测定牛奶和水样中四环素、盐酸土霉素和金霉素三种四环素类抗生素的分析方法。将0.1mmol盐酸土霉素在功能单体和交联剂的作用下制备分子印迹预聚合液,将经多巴胺处理后的不锈钢丝前端1~2cm置入其中,制备分子印迹固相微萃取涂层。3mL牛奶和水样经涂层萃取50min、解析5min,乙腈-10mmol/L磷酸盐缓冲液(PBS,pH=3)作为流动相,二极管阵列检测器(DAD)定量分析。实验结果表明,分子印迹涂层对四环素类目标物的特异性选择明显优于非印迹涂层。三种目标抗生素在100~1 000μg/L(牛奶)和10~1 000μg/L(水样)浓度范围内线性关系良好,相关系数在0.9959以上,四环素、盐酸土霉素和金霉素的检出限(S/N=3)为40~80μg/L(牛奶)和5~10μg/L(水样);加标水平为500μg/L时,回收率范围97.8%~109.0%,相对标准偏差(n=7)分别为5.3%~8.2%(牛奶),3.7%~6.4%(水样)。该方法前处理简单、绿色环保、选择性好、精密度好、回收率高,可用于牛奶和水样中上述三种四环素的实际检测。     

肌肉组织中四环素类抗生素的固相萃取-液相色谱法测定

建立了四环素类抗生素的高灵敏度测定方法，通过对实验条件的优化，采用固相萃取-反相高效液相色谱法同时测定肌肉组织中的土霉素(oxytetracycline)、四环素(tetracycline)、金霉素(chlortetracycline)3种四环素类抗生素残留量；土霉素、四环素、金霉素的检出限分别为20、40、100μg／kg；方法相对标准偏差为5．1％-7．1％(n＝5)，平均回收率为80％—82％。     

饲料中四环素类药物含量的高效液相色谱测定

建立了高效液相色谱法测定饲料中四环素类药物含量的方法。样品经甲醇-盐酸溶液提取,WatersOasis HLB固相萃取柱净化,进行HPLC分析。标准校正曲线线性范围为0.05~2.0 mg/L,相关系数r>0.999。饲料中土霉素、四环素、金霉素和强力霉素的定量下限(LOQ)依次是2.0、2.0、4.0和4.0 mg/kg;不同添加水平样品加标回收率为78%~103%,RSD小于11%。     

固相萃取-高效液相色谱法测定畜禽粪便中罗红霉素和3种四环素类抗生素

建立了固相萃取-高效液相色谱检测鸡粪和牛粪中的罗红霉素、土霉素、四环素和金霉素4种抗生素的分析方法。采用15 mL EDTA-McIlvaine缓冲液超声萃取15 min,萃取液浓缩后经HLB固相萃取柱净化,甲醇洗脱,洗脱液浓缩定容后用HPLC进行检测。实验结果表明,测定罗红霉素的线性范围为1.5~600 mg/L,土霉素、四环素和金霉素的线性范围为0.25~100 mg/L,相关系数为0.9995~0.9999。畜禽粪便中4种抗生素的检出限为0.06~0.38 mg/kg(S/N=3),定量限为0.2~1.2 mg/kg(S/N=10)。基质加标回收率为52.2%~64.7%,相对标准偏差为7.3%~8.4%。     

多模板分子印迹聚合物磁性固相萃取-高效液相色谱法测定环境水样中四环素类抗生素

以介孔硅磁性氧化石墨烯为载体,四环素、土霉素、金霉素、强力霉素共同作为模板,N-[3-(三甲氧基甲硅烷基)丙基]乙二胺(KH-792)和苯胺甲基三乙氧基硅烷(KH-42)为功能单体,通过溶胶-凝胶法制备了四环素类抗生素多模板分子印迹聚合物。以扫描电镜(SEM)、红外光谱(IR)及振动样品磁强计(VSM)对聚合物进行表征。将所制备的多模板分子印迹聚合物作为吸附剂应用于磁性固相萃取,结合高效液相色谱法,建立了水样中四环素、土霉素、金霉素及强力霉素测定的新方法。该方法对4种四环素类抗生素的线性范围为5～50μg/L,检出限为0.67～0.95μg/L,定量下限为2.13～3.50μg/L。实际样品的加标回收率为82.7%～103%,相对标准偏差(RSD)为1.0%～8.8%。方法可用于实际环境水样中4种四环素类抗生素的同时检测。     

模板分子印迹聚合物磁性固相萃取-高效液相色谱法测定环境水样中四环素类抗生素

以介孔硅磁性氧化石墨烯为载体,四环素、土霉素、金霉素、强力霉素共同作为模板,N-［3-(三甲氧基甲硅烷基)丙基］乙二胺(KH-792)和苯胺甲基三乙氧基硅烷(KH-42)为功能单体,通过溶胶-凝胶法制备了四环素类抗生素多模板分子印迹聚合物。以扫描电镜(SEM)、红外光谱(IR)及振动样品磁强计(VSM)对聚合物进行表征。将所制备的多模板分子印迹聚合物作为吸附剂应用于磁性固相萃取,结合高效液相色谱法,建立了水样中四环素、土霉素、金霉素及强力霉素测定的新方法。该方法对4种四环素类抗生素的线性范围为5~50 μg/L,检出限为0.67~0.95 μg/L,定量下限为2.13~3.50 μg/L。实际样品的加标回收率为82.7%~103%,相对标准偏差(RSD)为1.0%~8.8%。方法可用于实际环境水样中4种四环素类抗生素的同时检测。     

高效液相色谱-串联质谱法对化妆品中9种四环素类药物的同时测定

建立了化妆品中四环素、土霉素、金霉素、强力霉素、甲烯土霉素、去甲基金霉素、二甲胺四环素、脱水四环素和差相四环素含量的液相色谱串联质谱测定方法.对提取溶剂、流动相、流动相的梯度以及质谱参数进行了研究.采用乙腈-甲醇-草酸水溶液提取,LC-MS/MS分离测定,样品中被测物与干扰物能得到良好的分离.甲烯土霉素、去甲基金霉素在0.05 ～10.0 mg/L 范围内其质量浓度与峰面积均呈良好的线性关系,其余7种化合物在0.01 ～10.0 mg/L范围内其质量浓度与峰面积呈良好的线性关系.添加1.0 ～50.0 mg/kg时的加标回收率为73% ～99%,相对标准偏差低于13.2%.该法简便、快速、准确,可用于化妆品中上述9种四环素类药物的定性、定量检测.     

磁性固相萃取-高效液相色谱-串联质谱法测定环境水样中的四环素类抗生素

建立了磁性固相萃取-高效液相色谱-串联质谱同时测定环境水样中四环素类抗生素的方法。以6种四环素类抗生素(差向四环素、土霉素、四环素、去甲金霉素、金霉素和脱水四环素)为目标化合物,考察并优化了吸附和解吸条件,确定了最佳萃取条件。萃取后的目标化合物经ZORBAX Eclipse Plus C₁₈柱分离,用高效液相色谱-串联质谱在多反应监测(MRM)模式下进行检测。在优化的条件下,6种四环素在1~100 μg/L范围内线性关系良好,线性相关系数为0.9967~0.9993,检出限为2.44~25.21 ng/L,样品加标回收率为80.6%~90.0%,日内相对标准偏差(RSDs)为0.6%~2.5%,日间RSDs为1.1%~7.1%。该方法灵敏度高、背景干扰低,适用于环境水样中6种痕量四环素类抗生素的同时检测。     

高效液相色谱法和毛细管电泳法测定甘草制品中甘草酸的含量

分别用高效液相色谱法（ＨＰＬＣ）、毛细管区带电泳法（ＣＺＥ）、胶束电动毛细管色谱法（ＭＥＣＣ）测定了甘草制品中甘草酸的含量。对ＨＰＬＣ,ＣＺＥ,ＭＥＣＣ的分析条件作了一些选择实验,结果表明ＭＥＣＣ法与ＨＰＬＣ法分析数据接近、比较准确,而且前者比ＨＰＬＣ法分离效率高、溶剂用量少,是一种很有发展潜力的分析方法。     

Review. Use of laser detectors in capillary liquid chromatography

The main relationship of high-performance liquid chromatography (HPLC) are considered. It is shown that the optimum conditions of ultrasensitive trace analysis should be achieved by using packed capillary columns manufactured from flexible quartz capillaries with dc approximately less than 0.2 mm. The main features of these columns (v opt = 0.6 v opt of that for conventional HPLC columns with double the hydraulic permeability) make it possible to obtain two or three times higher plate numbers for the same analysis time and column pressure characteristic of conventional HPLC, as a result of using a submicrometre sorbent. The main features of laser detection in capillary liquid chromatography (laser-induced fluorescence and cross-beam thermal lens absorption detectors) are considered. The requirements that should be met by a modern capillary liquid chromatograph based on using flexible quartz capillary columns with a submicrometre sorbent and laser detectors are formulated. Examples of using these systems for femtomole and attomole analyses of biological samples (amino acids and prostaglandins) are given.     

毛细管电色谱-激发诱导荧光检测动物性食品中多肽类抗生素

以4-氟-7-硝基-2,1,3-苯并呋咱（NBD-F）为荧光试剂,建立了毛细管电色谱-激光诱导荧光（CEC-LIF）检测动物性食品中痕量杆菌肽、多粘菌素B和粘杆菌素等环状多肽抗生素的分析方法。在50 mmol/L的硼酸缓冲液（pH 7.5）中,多肽类抗生素经60℃衍生反应45 min。所得衍生产物采用苯基毛细管色谱填充柱进行分离,流动相为乙腈-磷酸盐缓冲液（pH 5.0,10 mmol/L）（55∶45,v/v）,辅助压力为3.8 MPa,分离电压为-10 kV,流速为0.02 mL/min。结果表明,多肽类抗生素的检出限（S/N=3）为5.0~10.0 ng/mL,满足目标物最大残留限量的检测要求。方法应用于饲料和牛奶样品的分析,平均回收率为72.9%~112.4%。该方法预处理操作简单、灵敏,为动物性食品中兽药残留分离分析提供了新手段。     

High-performance liquid chromatographic determination of loracarbef, a potential metabolite, cefaclor and cephalexin in human plasma, serum and urine

A high-performance liquid chromatographic (HPLC) method is reported for the determination of a new carbacephem antibiotic, loracarbef, a hydroxylated analogue, and two cephalosporins, cefaclor and cephalexin, in plasma, serum, and urine. The antibiotics are extracted from plasma by means of C18 solid-phase cartridges. Urine samples are diluted with water and directly injected on the HPLC system. The HPLC system utilizes a Supelcosil LC-18-DB (250 mm x 4.6 mm I.D.) reversed-phase column and ultraviolet detection at 265 nm. The limit of quantitation is 0.5 micrograms/ml for each compound. Excellent correlation of plasma concentrations is shown between results determined by HPLC and those obtained by microbiological agar-well diffusion assays. Stability studies of loracarbef in human plasma show the antibiotic to be stable for at least 24 h at room temperature and for at least twelve months at -20 degrees C.     

Solid-phase extraction of vinblastine and vincristine from plasma and urine: variable drug recoveries due to non-reproducible column packings

A sensitive and selective high-performance liquid chromatographic (HPLC) method for the determination of vinblastine and vincristine in plasma and urine is described. The drugs are isolated from 1.0 ml of the biological fluid with a solid-phase extraction column (Bond-Elut Diol). The HPLC method was combined with electrochemical detection at +850 mV versus an Ag/AgCl reference electrode. The detection limit is 100 pg for vinblastine and 250 pg for vincristine with a signal-to-noise ratio of 3, which permits the determination of these compounds in biological fluids at the nanogram level. Evaluation of the isolation method revealed that the drug recoveries and the reproducibility of the extraction procedure depend on the batch number of the solid-phase extraction column used.     

MicroHLPC determination of amygdalin in Semen pruni armeniacae and Semen pruni persicae

The application of micro HPLC to the determination of amygdalin in Semen pruni armeniacae and Semen pruni persicae is described. Amygdalin is separated at ambient temperature on a reversed phase column of U-Finepak SIL C18(150 x 0.5 mm) with methanol + water (25:75 v/v) as the mobile phase at a flow rate of 10 microL/min. The results are calculated by the internal standard method. The linear range is 1-7 micrograms. The CV and recovery of pure amygdalin are 1.47% (n = 10) and 98.13%, respectively. The results of analysis are lower than those obtained by TLC, but microHPLC is much simpler, faster, and more sensitive and reproducible than TLC.     

Trace determination of 10 β-lactam antibiotics in environmental and food samples by capillary liquid chromatography

A sensitive and reliable method using capillary HPLC with UV-diode array detection (DAD) has been developed and validated for the trace determination of residues of 10 β-lactam antibiotics of human and veterinary use, in milk, chicken meat and environmental water samples. The analytes included ampicillin, amoxicillin, penicillin V, penicillin G, cloxacillin, oxacillin, dicloxacillin, nafcillin, piperacillin and clavulanic acid. Legal levels are regulated by the EU Council regulation 2377/90 in animal edible tissues for these compounds. For food analysis, a solid-phase extraction (SPE) procedure consisting in a tandem of Oasis HLB and Alumina N cartridges was applied for off-line preconcentration and cleanup. For water analysis, the first step was only necessary. The limits of detection for the studied compounds were between 0.04–0.06 μg l⁻¹ for water samples and 0.80–1.40 μg l⁻¹ (or μg kg⁻¹) in the case of foods derived from animals. Average recoveries for fortified samples at different concentration levels ranged between 82.9% and 98.2%, with relative standard deviations (RSDs) lower than 9%. The method showed the advantages of capillary HPLC for the detection of these widely applied antibiotics in different samples at very low concentration levels.     

A sensitive and selective determination method of histamine by HPLC with intramolecular excimer-forming derivatization and fluorescence detection

A highly sensitive, selective and simple method is described for the determination of histamine by high-performance liquid chromatography (HPLC) with fluorescence detection. The method is based on an intramolecular excimer-forming fluorescence derivatization of histamine with 4-(1-pyrene)butyric acid N-hydroxysuccinimide ester (PSE), followed by reversed-phase HPLC. Histamine, having two amino moieties in a molecule, was converted to the dipyrene-labeled derivative by reaction with PSE. The derivative afforded intramolecular excimer fluorescence (450-540 nm), which can clearly be discriminated from the monomer fluorescence (370-420 nm) emitted from PSE. Typically, a 10 micro L sample solution was mixed with 100 micro L of derivatization reagent solution, which was a mixture of 0.5 mm PSE in acetonitrile and 0.5 mm potassium carbonate in water (8:2, v/v). The derivatization was carried out at 100 degrees C for 90 min. The PSE derivative of histamine could be separated by reversed-phase ODS column with isocratic elution using acetonitrile:water (82:18, v/v) containing 0.03% triethylamine. The detection limit (singnal-to-noise ratio = 3) of histamine was 0.5 fmol for a 30 micro L injection. The method was successfully applied to the determination of histamine in human urine, and had enough selectivity and sensitivity for urinary histamine quantification.     

Simultaneous determination of hydroxyanthraquinones in rhubarb and experimental animal bodies by high-performance liquid chromatography.

A simple and reliable high-performance liquid chromatographic (HPLC) method was developed for the simultaneous determination of five hydroxyanthraquinones (aloe-emodin, rhein, emodin, chrysophanol, and physcion) in Rhubarb and experimental animal bodies. A Zorbax SB-C18 column (250 mm x 4.6 mm i.d., 5 microm) and a methanol-0.5% acetic acid (85:15, v/v) mobile phase were used for the separation. The detection wavelength of a diode array detector (DAD) was set at 254 nm. Regression equations revealed a linear relationship (R2>0.9996) between the mass of hydroxyanthraquinones injected and the peak areas detected by DAD. The detection limits (S/N=3) ranged from 0.35 ng to 3.13 ng, and the recoveries ranged from 83% to 103% for different hydroxyanthraquinones. This method is simple, sensitive and suitable for the analysis of hydroxyanthraquinones in medicinal materials and pharmacological experiment samples.     

Determination of carbamates at trace levels in water and cucumber by capillary liquid chromatography

A sensitive and reliable method using capillary HPLC with UV-diode array detection (DAD) has been developed and validated for trace determination of carbamate pesticides in cucumber and environmental water samples. The analytes, including carbofuran, carbaryl, methiocarb, promecarb, benthiocarb and fenoxycarb, present legal residue levels regulated by the EU Council Directive 98/83/EC on drinking water and by the Regulation (EC) No. 396/2005 on vegetables. A previous off-line solid-phase extraction (SPE) procedure was required for preconcentration and sample clean-up. The separation was achieved using a C₁₈ column (150?mm?×?0.5?mm I.D, 5?µm particle size) and a mobile phase consisting of ACN?:?water using gradient mode, with a flow rate of 10?µl min^?1. Taking advantages of the characteristics of capillary HPLC, low volume of sample and solvents were required, achieving limits of detection for the studied compounds ranged from 10.0–29.6?ng l^?1 for water samples and 1.8–5.6?µg kg^?1 for cucumber, using UV-detection. Recoveries studies for fortified samples, at three different concentration levels, were carried out obtaining recoveries ranging from 70.0 to 111.1% and relative standard deviations (RSDs) lower than 10.6%.