尿样及动物组织中甲氧苄氨嘧啶残留量的高效液相色谱-串联质谱测定法

采用微量化样品前处理技术,以固相萃取为净化方法,电喷雾正离子多反应监测方式建立了尿样及动物组织中甲氧苄氨嘧啶残留量的液相色谱-串联质谱联用测定法.检出限为0.1 ng/g,线性范围均大于3个数量级,线性方程的相关系数大于0.999,组织样品和尿样的回收率分别为89%和93%.     

固相萃取-气相色谱-质谱联用法测定人尿中11种蛋白同化激素

建立了固相萃取（SPE）-气相色谱-质谱联用（GC-MS）同时测定尿样中11种蛋白同化激素的分析方法,并将该方法用于关雄酮（MADE）阳性尿的分析。尿样前处理采用含500mg填料的C18 SPE柱净化浓缩,然后进行酶解,最后由液-液萃取获得待测样品。为了提高灵敏度,待测样品采用硅烷化衍生后进行GC-MS分析。在选择离子监测（SIM）模式下,采用甲睾酮（MT）为内标进行定量分析,方法检出限为0．1～2．0μg·L^-1;分析加标实际样品,回收率在70．5％～121％,相对标准偏差为1．1％～13．9％。     

柱前荧光衍生-高效液相色谱法测定尿和血清中的环境雌激素

建立了同时测定人尿和血清中环境雌激素双酚A、4-壬基酚、17α-乙炔基雌二醇及内源性雌激素雌三醇、17α-雌二醇和17β-雌二醇的对硝基苯甲酰氯柱前荧光衍生-高效液相色谱测定方法。尿样经酸水解、固相萃取柱浓缩、净化分离;血清样品经乙腈沉淀蛋白后,用乙醚萃取游离态雌激素,N2气流挥干、在无水条件下,雌激素与对硝基苯甲酰氯反应生成荧光产物,用高效液相色谱法定量。检出限为2．7—8．3μg／L;加标回收率尿样为78．0％-102．5％,血清样为72．6％-98．6％;方法精密度为1．29％-4．52％。应用本法对20份尿样和10份血清样进行了测定,结果满意。     

固相萃取-气相色谱-质谱法测定猪组织中沙丁胺醇残留量

建立了动物组织中沙丁胺醇残留量固相萃取-气相色谱-质谱分析方法。动物组织样品经过葡萄糖醛甙酶酶解后调节pH至9．5，然后用异丙醇／乙酸乙酯混合溶剂液液分配去除杂质，旋转浓缩后用乙酸铵缓冲溶液溶解经SCX固相萃取（SPE）柱净化，洗脱液经氮气吹干后用双三甲基硅基三氟乙酰氨（BSTFA）衍生，采用选择离子模式（86、350、369、440）进行测定，外标法定量。检出限为0．10μg/kg。在添加浓度0．5～5．0μg/kg范围内，平均添加回收率在66．4％～82．4％，相对标准偏差（批内）（CV）在3．5％～5．9％之间；批间（CV）在2．2％～4．8％之间。衍生物的峰面积与样品浓度在0．002～0．50mg／L范围内呈良好的线性关系，线性回归系数大于0．999。     

固相萃取-气相色谱法测定动物组织中盐酸克伦特罗残留量

建立了固相萃取-气相色谱分析动物组织中盐酸克伦特罗残留量的方法．采用固相萃取,分离富集动物组织中的盐酸克伦特罗．肉样的加标回收率在70％～80％;最低检出限为1μg／kg．盐酸克伦特罗的硅烷化衍生产物,采用气相色谱(ECD检测器)检测,其衍生物的峰面积与样品浓度在0．005～1．0μg／mL范围内呈良好的线性关系,线性回归系数大于0．999．     

人工尿样中芥子气-谷胱甘肽加合物的气相色谱-质谱法检测

该文以气相色谱-质谱法测定人工尿样中的芥子气-谷胱甘肽加合物的β-裂解产物1,1′-磺酰基二(2-甲巯基)乙烷(SBMTE)。采用固相萃取(SPE)及固相支持液-液萃取(SLE)两种方法对尿样中的SBMTE进行富集净化,并对两种方法的影响因素进行了优化。采用中等极性毛细管色谱柱(DB-17MS)分离,以电子轰击源(EI)质谱选择离子模式(SIM)检测,内标法定量。结果表明,以SPE/GC-MS方法检测,SBMTE在1~100μg/L范围内线性关系良好,相关系数r为0.998 4,检出限(LOD)为0.1μg/L,回收率大于90%;以SLE/GC-MS方法检测,SBMTE在2~150μg/L范围内线性关系良好,相关系数r为0.998 8,检出限为0.5μg/L,回收率大于90%。SPE方法适用于直接检测空白尿样中添加SBMTE或芥子气染毒真实尿样还原后样品的进一步净化处理,SLE方法可应用于芥子气染毒后真实尿样的确证检测和溯源性检测。     

高效液相色谱法测定人尿中神经鞘氨醇和二氢神经鞘氨醇

建立了一种检测人尿中神经鞘氨醇（So）和二氢神经鞘氨醇（Sa）的高效液相色谱方法（HPLC）。离心分离尿样中的片状剥落细胞,裂解后用乙酸乙酯萃取、邻苯二甲醛衍生,在HPLC系统中通过梯度洗脱用Nova-PakC18－RP色谱柱（15cm×3．9mm,4μm）分离、荧光检测器检测。So和Sa的检出限均为0．05ng（女性尿样0,075μg/L、男性尿样0．005μg/L）。分析从我国一个村在采集的40份尿样,女性尿样中So、Sa和Sa／So比值分别为1．29－13．58μg/L、0．25－3．13μg/L和0．15－0．25,男性尿样中分别为0．075~3．07μg/L、0.019－0．50μg/L和0．028～0．26。     

柱切换高效液相色谱法在生物样品中的应用

评述了柱切换高效液相色谱法（ＣＳＨＰＬＣ）在生物样品分析中的应用进展．分别对柱切换装置，血样、尿样提取液中被测组分浓度的测定，在线浓缩样品技术以及ＣＳＨＰＬＣ的联用进行了介绍．     

奋乃静和氟奋乃静的毛细管电泳柱端喷壁式安培检测

应用自组装毛细管电泳．安培检测装置，建立了同时分离检测抗精神病药奋乃静与氟奋乃静的新方法。在1050mV(vs．Ag／AgCl)检测电位下，奋乃静与氟奋乃静的检出限分别达0．060mg／L与0．075mg／L。在最佳检测条件下，两组分的响应电流与浓度之间有着良好的线性关系，线性范围为0．75～75mg／L。将方法用于药物片剂以及模拟尿样和血清样品中奋乃静和氟奋乃静的测定，结果令人满意。     

稀土电解质Ce0.9M0.1O2-δ(M=Pr,Nd,Sm,Gd,Dy)的制备与性能

用溶胶-凝胶法制备中温电解质材料Ce0．9M0.1O2-δ,(M=Pr，Nd，Sm，Gd，Dy)系列样品。X射线衍射分析表明，样品为单相立方萤石结构，晶胞体积随着M原子序数的增加而减小。高温阻抗测量表明样品Ce0．9M0.1O2-δ电导率最高。Ce0．9M0.1O2-δ系列样品随着M原子序数的增加热膨胀系数减小。     

Liquid chromatography–electrospray ionization tandem mass spectrometry for the quantification of styraxlignolide A in rat plasma

Styraxlignolide A is a pharmacologically active ingredient isolated from Styrax japonica Sieb. et Zucc. A rapid, selective, and sensitive liquid chromatographic method with electrospray ionization tandem mass spectrometry was developed for use in the quantification of styraxlignolide A in rat plasma. Styraxlignolide A was extracted from rat plasma using ethyl acetate at neutral pH. The analytes were separated on an Atlantis dC₁₈ column using a mixture of methanol and ammonium formate (10 mM, pH 3.0) (70:30, v/v) and detected by tandem mass spectrometry in multiple reaction monitoring mode. The standard curve was linear (r²=0.9978) over the concentration range of 100?10000 ng/mL. The lower limit of quantification was 100 ng/mL using 50 μL of plasma sample. The coefficient of variation and relative error for intra‐ and inter‐assays at four QC levels were 1.6–8.3% and from ?12.0 to ?1.7%, respectively. The present method was applied successfully to the pharmacokinetic study of styraxlignolide A after intravenous administration of styraxlignolide A at a dose of 10 mg/kg in male Sprague–Dawley rats.     

In vivo quantification and pharmacokinetic studies of cotinine in mice after smoke exposure by LC–MS/MS

A sensitive analytical method was developed and validated for the quantification of cotinine in mouse plasma after exposure to smoke of 0.5, 1.0, and 1.5 commercially available cigarettes, using liquid chromatography tandem mass spectrometry. The method was validated over a linear concentration range of 0.075–20.0 ng/mL with the R² value being higher than 0.99. Both the precision (coefficient of variation; %) and accuracy (relative error; %) were within acceptable criteria of <15%. The lower limit of quantification (LLOQ) for cotinine was 0.075 ng/mL with sufficient specificity, accuracy, and precision. Following exposure to 0.5, 1.0, and 1.5 cigarette smoke, it was observed that the AUC and the C_max increased linearly as the doses increased. The pharmacokinetics of cotinine was found linear for the range of 0.5–1.5 commercial cigarette smoke. The quantification of the concentration of cotinine in mouse plasma after smoke exposure will facilitate future behavioral and toxicological experiments in animals and may prove useful in predicting cotinine levels in humans during smoking.     

Simultaneous and rapid determination of gefitinib,erlotinib and afatinib plasma levels using liquid chromatography/tandem mass spectrometry in patients with non‐small‐cell lung cancer

A simultaneous, selective, sensitive and rapid liquid chromatography/tandem mass spectrometry method was developed and validated for the quantification of gefitinib, erlotinib and afatinib in 250 μL samples of human blood plasma. Diluted plasma samples were extracted using a liquid‐phase extraction procedure with tert‐butyl methyl ether. The three drugs were separated by high‐performance liquid chromatography using a C₁₈ column and an isocratic mobile phase running at a flow rate of 0.2 mL/min for 5 min. The drugs were detected using a tandem mass spectrometer with electrospray ionization using imatinib as an internal standard. Calibration curves were generated over the linear concentration range of 0.05–100 nm in plasma with a lower limit of quantification of 0.01 or 0.05 nm for all compounds. Finally, the validated method was applied to a clinical pharmacokinetic study in patients with nonsmall‐cell lung cancer (NSCLC) following the oral administration of afatinib. These results indicate that this method is suitable for assessing the risks and benefits of chemotherapy in patients with NSCLC and is useful for therapeutic drug monitoring for NSCLC treatment. As far as we know, this is the first report on LC‐MS/MS method for the simultaneous quantification of NSCLC tyrosine kinase inhibitor plasma concentrations including afatinib. Copyright © 2015 John Wiley & Sons, Ltd.     

Simultaneous quantification of methylene blue and its major metabolite,azure B,in plasma by LC‐MS/MS and its application for a pharmacokinetic study

A simple and sensitive liquid chromatography‐tandem mass spectrometry (LC‐MS/MS) method was developed for the quantification of methylene blue (MB) and its major metabolite, azure B (AZB), in rat plasma. A simple protein precipitation using acetonitrile was followed by injection of the supernatant on to a Zorbax HILIC Plus column (3.5 µm, 2.1 × 100 mm) with isocratic mobile phase consisting of 5 mM ammonium acetate in 10:90 (v/v) water:methanol at a flow rate of 0.3 mL/min and detection in positive ionization mode. The standard curve was linear over the concentration range from 1 to 1000 ng/mL for MB and AZB with coefficient of determination above 0.9930. The lower limit of quantification was 1 ng/mL using 20 μL of rat plasma sample. The intra‐ and inter‐assay precision and accuracy were <12%. The developed analytical method was successfully applied to the pharmacokinetic study of MB and AZB in rats. Copyright © 2013 John Wiley & Sons, Ltd.     

Quantification of lurasidone, an atypical antipsychotic drug, in rat plasma with high-performance liquid chromatography with tandem mass spectrometry

A liquid chromatography–tandem mass spectrometric (LC/MS/MS) method was developed for the determination of an atypical antipsychotic drug, lurasidone, in rat plasma. The method involves the addition of acetonitrile and ziprasidone (internal standard) solution to plasma samples, followed by centrifugation. An aliquot of the supernatant was diluted with water and directly injected into the LC/MS/MS system. The separations were performed on a column packed with octadecylsilica (5 μm, 2.0 × 50 mm) with 0.1% formic acid and 0.1% formic acid in acetonitrile as mobile phase and the detection was performed using tandem mass spectrometry by multiple‐reaction monitoring via an electrospray ionization source. The standard curve was linear (r = 0.9982) over the concentration range 0.002–1 μg/mL. The intra‐ and inter‐assay precisions were 1.7 and 8.6%, respectively. The accuracy range was from 90.3 to 101.8%. The lower limit of quantification was 2.0 ng/mL using 50 μL of rat plasma sample. The developed analytical method was successfully applied to the pharmacokinetic study of lurasidone in rats. Copyright © 2011 John Wiley & Sons, Ltd.     

Determination of clopidogrel in human plasma by liquid chromatography/tandem mass spectrometry: application to a clinical pharmacokinetic study

A rapid and sensitive LC/MS/MS assay was developed and validated for the determination of clopidogrel in human plasma. Clopidogrel was extracted by single liquid-liquid extraction with pentane, and chromatographic separations were achieved on a C(18) column. The method was validated to demonstrate the specificity, linearity, recovery, lower limit of quantification (LLOQ), stability, accuracy and precision. The multiple reaction monitoring was based on m/z transition of 322.2 --> 211.9 for clopidogrel and 264.1 --> 125.1 for ticlopidine (internal standard). The total analytical run time was relatively short (3 min), and the LLOQ was 10 pg/mL using 0.5 mL of human plasma. The assay was linear over a concentration range from 10 to 10,000 pg/mL (r > 0.999). The intra- and inter-day accuracies were 101.3-108.8 and 98.4-103.5%, respectively, and the intra- and inter-day assay precisions were 1.9-5.5 and 4.4-8.1%, respectively. The developed assay method was applied to a pharmacokinetic study in human volunteers after oral administration of clopidogrel at a dose of 150 mg.     

Development of a rapid and sensitive liquid chromatography/tandem mass spectrometry method for the determination of 1,2-[bis(1,2-benzisoselenazolone-3(2H)-ketone)]-ethane (BBSKE), a novel anti-cancer agent in rat plasma

A rapid and sensitive liquid chromatography/electrospray ionization tandem mass spectrometric (LC/ESI-MS/MS) method has been developed to determine 1, 2-[bis(1,2-benzisoselenazolone-3(2H)-ketone)]-ethane (BBSKE), a novel antineoplastic agent, in rat plasma. The analytes were separated on a C18 column with a mobile phase of methanol-water (75:25, v/v) and detected using a triple-quadrupole mass spectrometer in positive mode with the selective reaction monitoring. The characteristic ion dissociation transitions were m/z 603.0 --> 448.9 for derivatized BBSKE and m/z 631.0 --> 476.8 for derivatized internal standard. The assay was linear over a range of 1-1000 ng/mL with a lower limit of quantification of 1 ng/mL. Intra- and inter-day precisions were less than 9.6 and 5.0%, respectively, and the accuracy ranged from -5.2 to 4.0%. The validated method was successfully applied to the characterization of pharmacokinetic profile of BBSKE after oral administration in rats. Cop     

HPLC-MS/MS method to determine genipin in rat plasma after hydrolysis with sulfatase and its application to a pharmacokinetic study

A sensitive high‐performance liquid chromatography–tandem mass spectrometry (LC‐MS/MS) method was developed for the quantification of genipin in rat plasma after hydrolysis with sulfatase. Genipin could not be detected directly as it could be transformed into other forms such as conjugated‐genipin immediately after administration. The conjugated genipin could be hydrolyzed by sulfatase to genipin. The conditions of hydrolysis were investigated. Genipin and the internal standard, peoniflorin (IS), were separated on a reversed‐phase column by gradient elution and detected using an electrospray ion source on a 4000 QTrap triple‐quadrupole mass spectrometer. The quantification was performed using multiple reaction monitoring with selected precursor‐product ion pairs of the transitions m/z 225.0 → 122.7 and m/z 479.1 → 449.1 for genipin and peoniflorin. The assay was linear over the concentration range of 1.368–1368 ng/mL, with correlation coefficients of 0.9989. Intra‐ and inter‐day precisions and accuracy were all within 15%. The lower limit of quantification was 1.368 ng/mL. The recoveries of genipin and peoniflorin were more than 53.3 and 51.2%. The highly sensitive method was successfully applied to estimated pharmacokinetic parameters of genipin following oral and intravenous administration to rats. The absolute bioavailability of genipin was 80.2% in rat, which is the first report. Copyright © 2011 John Wiley & Sons, Ltd.     

Detection and validated quantification of toxic alkaloids in human blood plasma--comparison of LC-APCI-MS with LC-ESI-MS/MS

Poisonings with toxic plants may occur after abuse, intentional or accidental ingestion of plants. For diagnosis of such poisonings, multianalyte procedures were developed for detection and validated quantification of the toxic alkaloids aconitine, atropine, colchicine, coniine, cytisine, nicotine and its metabolite cotinine, physostigmine, and scopolamine in plasma using LC-APCI-MS and LC-ESI-MS/MS. After mixed-mode solid-phase extraction of 1 ml of plasma, the analytes were separated using a C8 base select separation column and gradient elution (acetonitrile/ammonium formate, pH 3.5). Calibration curves were used for quantification with cotinine-d(3), benzoylecgonine-d(3), and trimipramine-d(3) as internal standards. The method was validated according to international guidelines. Both assays were selective for the tested compounds. No instability was observed after repeated freezing and thawing or in processed samples. The assays were linear for coniine, cytisine, nicotine and its metabolite cotinine, from 50 to 1000 ng/ml using LC-APCI-MS and 1 to 1000 ng/ml using LC-ESI-MS/MS, respectively, and for aconitine, atropine, colchicine, physostigmine, and scopolamine from 5 to 100 ng/ml for LC-APCI-MS and 0.1 to 100 ng/ml for LC-ESI-MS/MS, respectively. Accuracy ranged from -38.6 to 14.0%, repeatability from 2.5 to 13.5%, and intermediate precision from 4.8 to 13.5% using LC-APCI-MS and from -38.3 to 8.3% for accuracy, from 3.5 to 13.8%, for repeatability, and from 4.3 to 14.7% for intermediate precision using LC-ESI-MS/MS. The lower limit of quantification was fixed at the lowest calibrator in the linearity experiments. With the exception of the greater sensitivity and higher identification power, LC-ESI-MS/MS had no major advantages over LC-APCI-MS. Both presented assays were applicable for sensitive detection of all studied analytes and for accurate and precise quantification, with the exception of the rather volatile nicotine. The applicability of the assays was demonstrated by analysis of plasma samples from suspected poisoning cases.     

Liquid chromatography–tandem mass spectrometry method of loxoprofen in human plasma

A rapid, sensitive and selective liquid chromatography–electrospray ionization mass spectrometric method for the determination of loxoprofen in human plasma was developed. Loxoprofen and ketoprofen (internal standard) were extracted from 20 µL of human plasma sample using ethyl acetate at acidic pH and analyzed on an Atlantis dC₁₈ column with the mobile phase of methanol:water (75:25, v/v). The analytes were quantified in the selected reaction monitoring mode. The standard curve was linear over the concentration range of 0.1–20 µg/mL with a lower limit of quantification of 0.1 µg/mL. The coefficient of variation and relative error for intra‐ and inter‐assay at four quality control levels were 2.8–5.2 and 4.8–7.0%, respectively. The recoveries of loxoprofen and ketoprofen were 69.7 and 67.6%, respectively. The matrix effects for loxoprofen and ketoprofen were practically absent. This method was successfully applied to the pharmacokinetic study of loxoprofen in humans. Copyright © 2009 John Wiley & Sons, Ltd.