Quantification of vincristine and tariquidar by liquid chromatography-tandem mass spectrometry in mouse whole blood using volumetric absorptive microsampling for pharmacokinetic applications

A liquid chromatography-tandem mass spectrometry method was developed and validated for the simultaneous quantification of vincristine and tariquidar in 10 μL of mouse whole blood using volumetric absorptive microsampling devices. Samples were extracted from the devices and quantified against calibrators prepared in a human blood plasma matrix. Separation of vincristine and tariquidar was achieved using a Shimpack XR ODS III C18 stationary phase and H₂O and methanol mobile phase solvents containing 0.1% formic acid, running a gradient elution at a flow rate of 0.2 mL/min over 6.0 min. The method was linear up to 1200 ng/mL (R² > 0.99 for both analytes), with calibrator accuracy within ± 15% of the nominal concentrations and analyte coefficient of variance <15% for both vincristine and tariquidar. Pharmacokinetic assessment of both analytes was successfully applied in mice as both single-agent therapy and combination therapy over a 24-h period, and a 2.3-fold increase in vincristine drug exposure was observed in combination with tariquidar. This study validates the use of this approach for longitudinal analysis of drug exposure in animal studies.     

Development and validation of a hydrophilic interaction liquid chromatography-tandem mass spectrometry method for the quantification of regadenoson in human plasma and its pharmacokinetic application

Regadenoson, the first selective adenosine A_2A receptor agonist, is used to perform exercise stress test during radionuclide myocardial perfusion imaging. To detect the concentration of regadenoson in human plasma, a simple, fast, and sensitive tandem mass spectrometry method was established herein. Acetonitrile was used as a protein precipitation agent. Chromatographic separation was completed in 6.5 min using a BEH HILIC column (50 × 2.1 mm, 1.7 μm). The mobile phase consisted of 10 mmol/L ammonium acetate/acetonitrile (gradient elution). To quantify regadenoson and regadenoson-d3, an API 4000 mass spectrometry in multiple reaction monitoring mode with transitions of 391.3→259.2 and 394.3→262.2, respectively, was utilized. The calibration curve was linear in the range of 0.100–50.0 μg/L, and the intrabatch and interbatch precisions were <9.7% and <13.0%, respectively, and the accuracy was 2.0–6.9%. There was no apparent matrix effect for regadenoson or regadenoson-d3. The developed method was used to study the pharmacokinetic characteristics of regadenoson in healthy Chinese subjects.     

Application of liquid chromatography-tandem mass spectrometry method for the simultaneous quantitative analysis of propentofylline and its chiral metabolite M1 in rats

A sensitive and selective liquid chromatographic–electrospray ionization mass spectrometric method for the simultaneous determination of propentofylline and enantiomers of its active metabolite M1 in rat serum, cortex and hippocampus was developed and validated according to GLP procedures. Sample preparations were carried out by liquid–liquid extraction using diethyl ether after the addition of the internal standard (pentoxifylline). The dried residue was reconstituted in mobile phase and injected onto a Chiralpak AD column (10 µm, 250 × 4.6 mm i.d.). The limit of quantification for propentofylline in serum, cortex and hippocampus was set at 0.25 ng/mL and for enantiomers of its metabolite M1 at 1.25 ng/mL. The established LC/ESI‐MS/MS method has been successfully applied to an initial pharmacokinetic study of propentofylline and also to assessment of distribution of parent drug and enantiomers of its pharmacologically active metabolite M1 to cortex and hippocampus after intravenous administration of propentofylline to rats at a dose of 5 mg/kg. Copyright © 2010 John Wiley & Sons, Ltd.     

Bioequivalence evaluation of Florfenicol pharmaceutics in pigs using liquid chromatography-tandem mass spectrometry

The bioequivalence of two Florfenicol (FF) products in pigs was evaluated. A 2?×?2 crossover trial with a 14 days wash-out period was performed. The pigs were orally administered in a single dose (2?mg/kg b.w of FF). Serum samples were analyzed using a liquid chromatography-tandem mass spectrometry method. The limit of quantification was 0.1?ng/mL, and the calibration range was 1.0–100.0?ng/mL. Furthermore, intraday and interday were 5.43–9.09% and 6.23–9.68%, respectively. The areas under the curve (AUC_0–48) for the test product B of FF and reference product A were 7289.61?±?1750.44 and 6545.01?±?2766.25?h?×?ng/mL, respectively. The highest concentrations of FF in the serum (C_max) were 726.05?±?211.77 and 641.97?±?117.94?ng/mL. The mean retention times (MRT) was 7.91?±?1.98 and 7.76?±?2.89?h while the half-lives (T_1/2) were 4.07?±?1.71 and 4.99?±?3.30?h. From the analysis of variance results, the p values of C_max and AUC_0–∞ for the 90% confidence interval were 0.492 and 0.320 (p?>?0.05), respectively. A comparison between the test product and the reference product showed no significant difference. Both products showed bioequivalence after being administered in pigs.     

Development of a sensitive method for simultaneous determination of fluticasone propionate and salmeterol in plasma samples by liquid chromatography-tandem mass spectrometry

A new method for the fast simultaneous quantification of fluticasone propionate and salmeterol from plasma samples by liquid chromatography–tandem mass spectrometry, with adequate sensitivity for pharmacokinetic applications, was developed and validated. The chromatographic separation and mass‐spectrometric parameters were optimized for the retention and detection of the two compounds, despite quite different structures and properties. Two columns connected in series were used, cation‐exchange (Zorbax 300‐SCX, 5 cm × 2.1 mm, 5 µm) and octadecyl (Discovery HSC₁₈, 10 cm × 2.1 mm, 5 µm). The mass‐spectrometric interface was operated in negative electrospray ionization mode; high sensitivity and lesser matrix effects were obtained, permitting smaller consumption of plasma. The sample preparation was based on supported liquid–liquid extraction in 96‐well format plates that provided clean samples with a simplified procedure that was suitable for automation. The method was validated according to regulatory guidelines, by assessing lower limits of quantification, selectivity, linearity, accuracy, precision, extraction recoveries and matrix effects. A comparison with two other methods for the separate determination of fluticasone propionate and salmeterol in plasma samples, previously developed by our group, is presented. The statistical evaluation of the results obtained with the three methods on a set of unknown samples from treated patients demonstrated good correlation (R² 0.987 for fluticasone propionate and 0.967 for salmeterol). Copyright © 2011 John Wiley & Sons, Ltd.     

Cellular toxicity and pharmacokinetic study of butachlor by ultra-performance liquid chromatography-tandem mass spectrometry based on tandem mass spectrometry cubed technique

Butachlor is an aromatic amide compound that plays a role as a herbicide, a xenobiotic, and an environmental contaminant. The aim of this work was to develop a highly selective and sensitive ultra-performance liquid chromatography-tandem mass spectrometry method based on the tandem mass spectrometry cubed technique to determine butachlor in a biological matrix. Butachlor and internal standard acetochlor were separated on a Waters Acquity ultra-performance liquid chromatography BEH C₁₈ column (2.1 × 50 mm, 1.7 μm) with gradient elution using 0.1% formic acid aqueous solution (A) and acetonitrile (B) as mobile phases. The transitions selected for tandem mass spectrometry cubed quantitative analysis in positive ion mode were: for butachlor, mass-to-charge ratio 312.2→238.1→162.1; for acetochlor, mass-to-charge ratio 270.1→224.0→148.1. The total running time for each sample was 5.5 min. The ultra-performance liquid chromatography-tandem mass spectrometry cubed method showed a linear relationship (R² ≥ 0.995) in the concentration range of 0.5–100 ng/ml. The intra and interday accuracies are within the range of -10.6%–4.3% and precisions are between 4.48% and 13.14%. The novelty of the method is the use of tandem mass spectrometry cubed scanning mode, which improves selectivity and sensitivity. The results indicated that butachlor was cellular toxic. The safety of butachlor should be considered when it is used as a herbicide.     

高效液相色谱-串联质谱法测定比格犬血浆中的莫诺苷浓度及其药代动力学

运用高效液相色谱-串联质谱联用技术,建立了简单、快速、灵敏的比格犬灌胃莫诺苷后血药浓度的检测方法。血浆样品采用蛋白质沉淀法处理,以芍药苷作为内标,色谱柱为Inertsil ODS-SP色谱柱(50 mm×2.1 mm,5μm),流动相为水(含1 mmol/L甲酸钠)-乙腈,梯度洗脱,流速0.4 mL/min。采用电喷雾离子源(ESI),正离子多反应监测(MRM)模式。绘制血药浓度-时间曲线,并采用DAS 2.0软件计算药代动力学参数。方法学实验结果表明内源性杂质不干扰莫诺苷和内标的测定,线性范围为2~5 000μg/L(r=0.996 6),定量限为2μg/L。方法精密度、准确度、回收率和基质效应均符合生物样品测定的要求,适合比格犬血浆中莫诺苷浓度的测定,可以应用该方法进行莫诺苷的药代动力学研究。比格犬灌胃莫诺苷3个剂量(5、15、45 mg/kg)后的血药浓度-时间曲线下面积(AUC(0-∞))分别为(1 631.20±238.50)、(3 984.05±750.38)、(10 397.64±3 156.34)μg/L·h,与给药剂量之间呈现良好的线性关系。     

毛细管反相液相色谱-串联质谱用于肽的鉴定和相对定量分析

建立了一种基于毛细管反相液相色谱-串联质谱联用技术和质谱峰强度数据处理的肽段鉴定和相对定量分析方法。该方法无需对样品中的肽进行化学标记,在对样品进行反相色谱分离和串联质谱分析后,将二级质谱扫描数据进行蛋白质数据库搜索,获得所鉴定肽段的序列、保留时间、质荷比、带电荷数等定性信息;再以此为定位依据,在全扫描质谱数据中提取该肽段对应的离子峰并以该离子峰的峰强度作为定量信息,从而实现对不同样品中的共有肽段进行差异比较分析。以标准蛋白酶解混合肽段为实验对象,以肽段相对强度的相对标准偏差为指标,考察了该方法用于肽段相对定量分析的重现性、检测动态范围以及浓度标准曲线等,为将该方法用于生物样品中内源性肽的差异分析奠定了基础。     

超高效液相色谱-串联质谱法测定人血浆中苯海索

建立了灵敏、高通量的超高效液相色谱-串联质谱（UPLC-MS/MS）定量测定人血浆中苯海索的方法,用于盐酸苯海索片生物等效性研究,并确证食物对苯海索体内药代动力学行为的影响。以甲醇为沉淀剂进行蛋白质沉淀,苯海索-d11为内标,采用Waters Acquity UPLC BEH C8色谱柱（50 mm×2.1mm,1.7 μm）,以0.1%（v/v）甲酸水溶液（含5 mmol/L醋酸铵）和乙腈-水（95：5,v/v）为流动相进行梯度洗脱;采用电喷雾电离（ESI）源,在正离子模式下进行多反应监测（MRM）扫描。苯海索在0.1~40 ng/mL范围内线性关系良好。应用该方法测定中国健康受试者空腹及餐后单次口服2 mg盐酸苯海索片后的血药浓度,结果显示最大血药浓度（C_max）、血药浓度-时间曲线下面积（AUC_0-t、AUC_0-∞）的90%置信区间均在80.0%~125.0%范围内,表明两种制剂在空腹和餐后均生物等效。     

液相色谱-串联质谱法测定毛发中的司坦唑醇

采用液相色谱-串联质谱(LC-MS/MS)建立了毛发中司坦唑醇的分析方法,并将其应用于单次给药动物实验中的动物毛样品分析。将10 mg样品碱水解后加入戊烷提取,然后进行LC-MS/MS分析,采用正离子电喷雾电离、多反应监测模式测定,方法的最低定量限为25 pg/mg。剃去豚鼠背部中央的毛,以60 mg/kg的剂量于豚鼠腹腔注射司坦唑醇,然后隔天在同一部位剃取其毛,两周内该豚鼠毛中均能检出司坦唑醇;给药后豚鼠毛中司坦唑醇的含量在第1周内保持稳定,在给药后第10天达到峰值。所建立的方法毛发取样量少,特异性强,灵敏度高,适用于毛发中司坦唑醇的分析。     

高效液相色谱-串联质谱法检测牛奶中多种苯并咪唑兽药残留

采用正交试验优化的QuEChERS方法处理样品,建立了牛奶中包括前体药物和代谢物在内12种苯并咪唑类(BZs)兽药残留的高效液相色谱-串联质谱检测方法.方法在1～500μg/kg范围内线性良好,相关系数为0.9980～0.9996,平均回收率72.4%～108.3%,RSD为1.4%～9.7%,检出限低于0.5μg/k...     

超高效液相色谱-串联四极杆飞行时间质谱和超高效液相色谱-串联三重四极杆质谱用于血浆中苦杏仁苷及其代谢产物野黑樱苷的定性和定量分析

建立了大鼠灌胃麻杏石甘汤后血浆中苦杏仁苷、野黑樱苷的定性及定量方法。样品经液液萃取净化处理,定性采用超高效液相色谱-串联四极杆飞行时间质谱仪（UPLC-QTOF-MS/MS）,经Shim-pack XR-ODS Ⅲ色谱柱（75 mm×2.0 mm,1.6 μm）分离,定量采用超高效液相色谱-串联三重四极杆质谱仪（UPLC-Q-TRAP-MS）,经Agilent C₁₈色谱柱（50 mm×2.1 mm,1.7 μm）分离,电喷雾负离子化（ESI）及MRM模式测定,流动相均为乙腈-0.1%（v/v）甲酸水溶液。结果显示苦杏仁苷、野黑樱苷在相应浓度范围内线性关系良好（相关系数分别为0.9990、0.9970）,精密度（RSD）小于9.20%,回收率为82.33%~95.25%,检出限（LOD）约为0.50 ng/mL。本方法快速简便,为血浆样品中苦杏仁苷、野黑樱苷的定性和定量分析提供良好参考。     

高效液相色谱-串联质谱法测定盐酸二甲双胍及其制剂中痕量N-亚硝基二甲胺

建立了高效液相色谱-串联质谱(HPLC-MS/MS)检测盐酸二甲双胍原料和制剂中N-亚硝基二甲胺(NDMA)含量的方法。样品以水为提取溶剂,经涡旋混匀、恒温振荡、高速离心、微孔过滤后进行HPLC-MS/MS分析。采用ACE EXCEL 3 C18-AR色谱柱(150 mm×4.6 mm, 3μm)分离,流动相为均含0.1%甲酸的水和甲醇溶液,梯度洗脱,流速0.8 mL/min,柱温40℃,自动进样器温度10℃。采用阀切换技术保护质谱仪,设置六通阀切换使保留时间2.85～7.00 min的流动相进入质谱,其余时间流动相进入废液。质谱部分采用大气压化学电离(APCI)源,在正离子、MRM模式下扫描,雾化器流量为3 L/min,加热器流量为10 L/min,接口温度为300℃,脱溶剂管温度为250℃,加热块温度为400℃,干燥器流量为10 L/min。NDMA定量离子对为m/z 75.0→43.1,碰撞能量(CE)为-17.0 eV,定性离子对为m/z 75.0→58.2, CE为-16.0 eV。采用外标法定量。对方法进行了详细的方法学验证,结果表明,该法专属性良好,溶剂和辅料对NDMA测...     

Simultaneous quantification of fatty acids in serum with dual derivatization by liquid chromatography-tandem mass spectrometry

Fatty acids have multitudinous biological functions and play a crucial role in many biological processes, but due to poor ionization efficiency and lack of appropriate internal standards, the comprehensive quantification of fatty acids by liquid chromatography-tandem mass spectrometry is still challenging. In this study, a new, accurate, and reliable method for quantifying 30 fatty acids in serum using dual derivatization was proposed. Indole-3-acetic acid hydrazide derivants of fatty acids were used as the internal standard and indole-3-carboxylic acid hydrazide derivants of them were used to quantify. The derivatization conditions were systematically optimized and the method validation results showed good linearity with R² > 0.9942, low detection limit (0.03–0.6 nM), precision (1.6%–9.8% for intra-day and 4.6%–14.1% for inter-day), recovery (88.2%–107.2% with relative standard deviation < 10.5%), matrix effect (88.3%–105.2% with the relative standard deviation < 9.9%) and stability (3.4%–13.8% for fatty acids derivants in 24 h at 4°C and 4.2%–13.8% for three freeze-thaw cycles). Finally, this method was successfully applied to quantify fatty acids in serum samples of Alzheimer's patients. In contrast to the healthy control group, nine fatty acids showed a significant increase in the Alzheimer's disease group.     

液相色谱-串联质谱法测定水环境中的十氯酮

建立了分析测定水环境中十氯酮的液相色谱-串联质谱法。水样经液液萃取、净化后,采用Eclipse plus C18柱（100 mm×2.1 mm,3.5 μm）分离,乙腈和水为流动相进行梯度洗脱,在电喷雾负离子多反应监测模式下进行检测,同位素内标法定量。结果表明：采用液相色谱-质谱联用技术,证实了十氯酮在甲醇中以半缩醛的形式存在,而在丙酮/乙腈中以偕二醇的形式存在。由于十氯酮极性较强,在净化时难以洗脱,并且不耐酸,所以不能与其他有机氯农药一起分析。十氯酮在5~100 μg/L范围有良好的线性关系,相关系数r²=0.999,检出限及定量限分别为0.70 ng/L和2.8 ng/L;在5、40和100 ng/L 3个浓度添加水平的平均回收率为95.1%~98.9%,相对标准偏差为3.85%~4.72%。本方法具有良好的灵敏度、回收率和重现性,适用于水环境中十氯酮的测定。     

液相色谱-串联四极杆飞行时间质谱和超高效液相色谱-串联三重四极杆质谱用于检测牛肉中的刚果红

建立了牛肉中刚果红的检测方法。定性方法采用液相色谱-串联四极杆飞行时间质谱对未知物进行质谱谱图库匹配,定量分析采用超高效液相色谱-串联三重四极杆质谱。牛肉样品中的刚果红经液液萃取净化后,采用Agilent ZORBAX Eclipse Plus C₁₈ Rapid Resolution HD色谱柱(50 mm×2.1 mm, 1.8 μm)进行分离,流动相为95%(体积分数)甲醇,流速为0.2 mL/min。AB 4000⁺三重四极杆质谱仪在电喷雾负离子化(ESI)及MRM模式下定量。结果显示,刚果红在0.03~1 mg/L浓度范围内,线性关系良好(相关系数为0.9998),精密度良好(RSD小于5%),回收率为88%~91%,检出限约为0.01 mg/L。本方法快速简便,重现性好,可以为牛肉及其他肉制品中刚果红的定量提供良好的解决方案。     

超高效液相色谱-串联质谱法同时快速筛查检测食品中的10种抗凝血类鼠药

创建了一种利用超高效液相色谱-串联质谱同时快速筛查检测食品中10种抗凝血类鼠药的方法。试样用乙腈提取,QuEChERS净化,以5 mmol/L乙酸铵和乙腈为流动相进行梯度洗脱,经Poroshell 120 EC-C18柱(100 mm×2.1 mm, 2.7 μm;美国Agilent公司)分离,采用电喷雾电离(ESI)负离子质谱在多反应监测(MRM)模式下检测。10种鼠药均在5～500 μg/L范围内呈现良好的线性关系(r>0.99)。在辣椒酱、面粉、醋和酱油4种样品中,10种鼠药的加标回收率均在72.6%～112%范围内,相对标准偏差均不大于11.2%,检出限均在0.5～4.5 μg/kg范围内。本方法快速、简便、灵敏、重现性好,能满足由此10种抗凝血类鼠药引发的突发性公共卫生安全事件的快速筛查与检测要求。     

Sensitive liquid chromatography tandem mass spectrometry method for the quantification of Quetiapine in plasma

A sensitive high-performance liquid chromatography-tandem mass spectrometry method was developed and validated for the quantification of quetiapine in rat plasma. Following liquid-liquid extraction, the analyte was separated using a gradient mobile phase on a reverse-phase column and analyzed by MS/MS in the multiple reaction monitoring mode using the respective [M + H]+ ions, m/z 384 to m/z 221 for quetiapine and m/z 327 to m/z 270 for the internal standard. The assay exhibited a linear dynamic range of 0.25-500 ng/mL for quetiapine in rat plasma. The lower limit of quantification was 0.25 ng/mL with a relative standard deviation of less than 7%. Acceptable precision and accuracy were obtained for concentrations over the standard curve range. The validated method was successfully used to analyze rat plasma samples for application in pre-clinical pharmacokinetic studies. This method in rodent plasma could be adapted for quetiapine assay in human plasma.     

液相色谱-串联质谱法检测食品中的多种易滥用着色剂

建立了硬糖、果酱、液态奶、果汁中酸性红52、红色2G、喹啉黄、专利蓝、酸性红26、柠檬黄、靛蓝、胭脂红、诱惑红、日落黄、亮蓝、苋菜红等12种易滥用着色剂残留量的液相色谱-串联质谱分析方法。样品用水溶液稀释提取,经聚酰胺固相萃取柱净化后,在Agilent XDB-C18色谱柱分离,以20 mmol/L乙酸铵溶液和乙腈为流动相进行梯度洗脱。质谱采用电喷雾负离子(ESI~)模式电离,多反应监测(MRM)模式检测,基质匹配标准溶液外标法定量。易滥用着色剂在0.5～50 mg/L范围内呈线性相关,相关系数(r)均大于0.99,其定量限(信噪比大于10)为0.5 mg/kg,检出限(信噪比大于3)为0.1 mg/kg。各种基质样品在0.5、5和50 mg/kg添加水平时,易滥用着色剂的回收率范围为62.6%～115.3%,相对标准偏差(RSDs)为2.6%～26.3%,可以满足食品中易滥用着色剂含量的检测需要。     

Ultra‐performance liquid chromatography tandem mass spectrometry method for the determination of AZ66, a sigma receptor ligand,in rat plasma and its application to in vivo pharmacokinetics

Methamphetamine abuse continues as a major problem in the USA owing to its powerful psychological addictive properties. AZ66, 3‐[4‐(4‐cyclohexylpiperazine‐1‐yl)pentyl]‐6‐fluorobenzo[d]thiazole‐2(3H)‐one, an optimized sigma receptor ligand, is a promising therapeutic agent against methamphetamine. To study the in vivo pharmacokinetics of this novel sigma receptor ligand in rats, a sensitive ultra‐performance liquid chromatography/tandem mass spectrometry (UPLC/MS/MS) method was developed in rat plasma and validated. The developed method requires a small volume of plasma (100 μL) and a simple liquid–liquid extraction. The chromatographic separations were achieved in 3.3 min using an Acquity UPLC BEH Shield RP₁₈ column. The mass spectrophotometric detection was carried out using a Waters Micromass Quattro MicroTM triple‐quadrupole system. Multiple reaction monitoring was used for the quantitation with transitions m/z 406 → m/z 181 for AZ66 and m/z 448 → m/z 285 for aripiprazole. The method was validated over a concentration range of 1–3500 ng/mL and the lower limit of quantitation was determined to be 1 ng/mL. Validation of the assay demonstrated that the developed UPLC/MS/MS method was sensitive, accurate and selective for the determination of AZ66 in rat plasma. The present method has been successfully applied to an i.v. pharmacokinetic study in Sprague–Dawley rats. Copyright © 2013 John Wiley & Sons, Ltd.