LC-MS／MS法测定人血浆中氯沙坦及其代谢物E-3174的浓度

目的建立LC-MS／MS法测定人血浆中氯沙坦及其代谢物E-3174血药浓度的方法。方法血浆酸化后用乙醚提取,采用同位素内标（氘3-B3174）进行测定。色谱柱：CAPCELLPACKC18Ⅲ（100mm×2．0mm,5μm）,流动相：0．02％甲酸乙腈-水溶液（53：47,v／v）;等度洗脱;流速0．3mL·min-1;进样体积5μL;电喷雾离子化,正离子MRM扫描。结果氯沙坦和E-3174线性范围均为5—500μg·L-1（r〉0．999）,最低定量限均为5μg·L-1,平均提取回收率均〉50％,批内、批间精密度RSD均〈8％。结论本方法灵敏度高、专一性好、操作简单,适用于氯沙坦的药动学研究。     

LC-MS/MS法测定血浆中甲氯噻嗪的浓度

目的:建立血浆中甲氯噻嗪的LC-MS/MS定量测定方法.方法:以双氢克尿噻为内标,血浆样品经碱化后用含5%异丙醇的乙酸乙酯溶液提取,采用液质联用色谱法进行MRM扫描分析,色谱柱为CAPCELL PAK C18 MG(50mm×2.0mm,5μm),流动相为20mmol/L乙酸胺水-乙腈(64:36),离子选择通道分别为甲氯噻嗪:358.2/321.9amu;双氢克尿噻:296.3/268.9amu.甲氯噻嗪、双氢克尿噻的保留时间分别为1.6min和1.0min.结果:本文所建立的血浆样品中甲氯噻嗪液质联用色谱测定方法,血浆内源性物质不干扰样品峰,相对回收率为93.7%～105.4%;绝对回收率为71.2%～78.4%;日间和日内相对标准差均小于6.04%.血浆中的最低定量限为0.2ng/mL(S/N≥20:1),线性范围为0.2ng/mL～51.2ng/mL.结论:本方法操作简便,特异性强,灵敏度高,取血量少,符合生物样品的分析要求,可以用于甲氯噻嗪药代动力学研究及临床测定.     

Analysis of carvedilol in human plasma using hydrophilic interaction liquid chromatography with tandem mass spectrometry

A rapid, sensitive and selective method for the determination of carvedilol in human plasma was developed using hydrophilic interaction liquid chromatography with tandem mass spectrometry (HILIC-MS/MS). Carvedilol and cisapride (internal standard) were extracted from human plasma with methyl tert-butyl ether at basic pH and analyzed on an Atlantis HILIC Silica column with the mobile phase of acetonitrile-ammonium formate (50 mM, pH 4.5) (90:10, v/v). The analytes were detected using an electrospray ionization tandem mass spectrometry in the multiple-reaction-monitoring mode. The standard curve was linear (r=0.9998) over the concentration range of 0.1-200 ng/ml. The lower limit of quantification for carvedilol was 0.1 ng/ml using 50 microl plasma sample. The coefficient of variation and relative error for intra- and inter-assay at four QC levels were 1.6-4.5% and -6.4 to 4.8%, respectively. The absolute and relative matrix effect for carvedilol and cisapride were practically absent. The extraction recoveries of carvedilol and cisapride were 81.6 and 85.2%, respectively. This method was successfully applied to the bioequivalence study of carvedilol in humans.     

Method development and validation of almotriptan in human plasma by HPLC tandem mass spectrometry: application to pharmacokinetic study

A simple, sensitive and selective method has been developed for quantification of Almotriptan (AL) in human plasma using Almotriptan-d(6) (ALD6) as an internal standard. Almotriptan and Almotriptan-d(6) were detected with proton adducts at m/z 336.1→201.1 and 342.2→207.2 in multiple reaction monitoring (MRM) positive mode, respectively. The method was linear over a concentration range of 0.5-150.0 ng/mL. The limit of detection (LOD) and limit of quantification (LOQ) for Almotriptan were 0.2 pg/mL and 0.5 ng/mL, respectively. Liquid-liquid extraction was used followed by MS/MS (ion spray). The method was shown to be precise with an average within-run and between-run variation of 0.68 to 2.78% and 0.57 to 0.86%, respectively. The average within-run and between-run accuracy of the method throughout its linear range was 98.94 to 102.64% and 99.43 to 101.44%, respectively. The mean recovery of drug and internal standard from human plasma was 92.12 ± 4.32% and 89.62 ± 6.32%. It can be applied for clinical and pharmacokinetic studies.     

液相色谱-质谱／质谱联用法测定健康志愿者血清中阿德福韦

目的:建立测定人血清中阿德福韦的液相色谱-质谱/质谱联用(LC～MS/MS)方法。方法:血清样品经甲醇沉淀蛋白,上清液吹干,200μL流动相复溶,离心,取40μL进样。色谱柱为 Diamonsil C_(18)柱(250 mm×4.6 mm,5μm),流动相为甲醇-水-甲酸(20:80:0.1,v/v/v),流速0.6 mL·min~(-1),采用电喷雾离子化四极杆串联质谱,多反应监测方式测定样品浓度。监测离子对分别为 m/z274→m/z162(阿德福韦)和 m/z226→m/z135(内标阿昔洛韦)。结果:阿德福韦在1.25～160μg·L~(-1)浓度范围内线性关系良好(r=0.9992,n=5),最低定量限为1.25μg·L~(-1)。低、中、高3种浓度质控样品的日内、日间精密度小于8.64%,方法回收率99.20%～101.98%,阿德福韦提取回收率56.50%～59.26%。结论:该方法灵敏度高,定量准确,适用于阿德福韦酯人体药代动力学研究。     

Simultaneous determination of irbesartan and hydrochlorothiazide in human plasma using HPLC coupled with tandem mass spectrometry: Application to bioequivalence studies

A sensitive, specific and selective liquid chromatography/tandem mass spectrometric method has been developed and validated for the simultaneous determination of irbesartan and hydrochlorothiazide in human plasma. Plasma samples were prepared using protein precipitation with acetonitrile, the two analytes and the internal standard losartan were separated on a reverse phase C₁₈ column (50 mm × 4 mm, 3 μm) using water with 2.5% formic acid, methanol and acetonitrile (40:45:15, v/v/v (%)) as a mobile phase (flow rate of 0.70 mL/min). Irbesartan and hydrochlorothiazide were ionized using ESI source in negative ion mode, prior to detection by multiple reaction monitoring (MRM) mode while monitoring at the following transitions: m/z 296 → 269 and m/z 296 → 205 for hydrochlorothiazide, 427 → 175 for irbesartan. Linearity was demonstrated over the concentration range 0.06–6.00 μg/mL for irbesartan and 1.00–112.00 ng/mL for hydrochlorothiazide. The developed and validated method was successfully applied to a bioequivalence study of irbesartan (300 mg) with hydrochlorothiazide (12.5 mg) tablet in healthy volunteers (N = 36).     

Development and full validation of a sensitive quantitative assay for the determination of clemastine in human plasma by liquid chromatography-tandem mass spectrometry

A sensitive high-performance liquid chromatography-tandem mass spectrometry (LC/MS/MS) method was developed and validated for the determination of clemastine in human plasma. After having been extracted from plasma samples by ethyl acetate, clemastine and internal standard, diphenhydramine, were separated on a C(18) column. Detection was performed on Thermo Finnigan TSQ Quantum triple quadrupole mass spectrometer by selected reaction monitoring (SRM) mode via electrospray ionization (ESI) source. The method was linear in the concentration range of 5.0-1000.0 pg/ml for clemastine. The intra- and inter-day precisions were within 13.4% and the deviations were between -1.1% and 5.6%. The fully validated LC/ESI-MS/MS method has been successfully applied to the preliminary pharmacokinetic study in healthy male Chinese volunteers.     

Determination of lovastatin in human plasma by ultra-performance liquid chromatography-electrospray ionization tandem mass spectrometry and its application in a pharmacokinetic study

A selective, rapid and sensitive ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed for the quantitative determination of lovastatin in human plasma and its application in a pharmacokinetic study. With mycophenolate mofetil as internal standard, sample pretreatment involved a one-step extraction with tert-butyl methyl ether of 0.2 ml plasma. The analysis was carried out on an ACQUITY UPLCTM BEH C18 column (50 mm x 2.1 mm, i.d., 1.7 microm) with flow rate of 0.35 ml/min. The mobile phase was 20% water and 80% acetonitrile (v/v). The detection was performed on a triple-quadrupole tandem mass spectrometer by multiple reaction monitoring (MRM) mode via electrospray ionization (ESI). Linear calibration curves were obtained in the concentration range of 0.08-24.50 ng/ml, with a lower limit of quantification of 0.08 ng/ml. The intra- and inter-day precision (RSD) values were below 15% and accuracy (RE) was -7.6 to 9.3% at all QC levels. The method was applicable to clinical pharmacokinetic study of lovastatin in healthy volunteers following oral administration.     

Simultaneous determination of alfentanil and midazolam in human plasma using liquid chromatography and tandem mass spectrometry

A fast, sensitive and selective liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the determination of alfentanil and midazolam in human plasma has been developed and validated. Alfentanil and midazolam were extracted from plasma using a mixed-mode cation exchange solid phase extraction method, with recoveries of both compounds greater than 80% at 3 different concentrations (1, 10 and 100ng/ml). Compounds were analyzed on a C(18) column with a water and methanol mobile phase gradient with acetic acid as an additive, at a flow rate of 0.3ml/min. The working assay range was linear from 0.25 to 100ng/ml for each compound. The signal to noise ratio was 80 and 40 for alfentanil and midazolam, respectively, at the lowest concentration calibration standard, with less than 10% matrix suppression by human plasma at this concentration. Alfentanil and midazolam were stable in human plasma during storage at -80°C, processing, and analysis. The procedure was validated and applied to the analysis of plasma samples from healthy human subjects administered oral and intravenous alfentanil and midazolam.     

液相色谱-串联质谱法测定片剂中卡马西平的含量

目的 建立液相色谱-串联质谱（LC/MS/MS）测定片剂中的卡马西平含量的方法.方法 色谱柱Zorbax Extend-C18柱（150 mm×4.6 mm,5 μm）;流动相：甲醇-0.01 mmol/l乙酸胺溶液（80：20,v/v）;流速：0.3 ml/min.结果 卡马西平浓度在0.05～40 μg/,ml范围内,峰面积与浓度线性关系良好,平均回收率为100.2%,RSD为1.02%.日内精密度、日间精密度的RSD分别为1.72%和1.91%.4批样品中卡马西平的含量分别为96.3%、98.6%、96.5%、99.0%.结论 本方法具有良好的灵敏度、准确度、精确度及专属性,结果准确,重现性好,易于操作,可用于制剂中卡马西平含量的测定.     

Liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for simultaneous determination of tenofovir and emtricitabine in human plasma and its application to a bioequivalence study

A rapid and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method has been developed and validated for simultaneous quantification of Tenofovir (TEN) and Emtricitabine (EMT) in human plasma using Chromolith Speed Rod RP18. The mass transition ion-pair has been followed as m/z 288.10-->176.10 for TEN, m/z 248.20-->130.20 for EMT and m/z 230.10-->112.10 for Lamivudine (LAM). The method involves solid phase extraction from plasma, simple isocratic chromatographic conditions and mass spectrometric detection using an API 5000 instrument that enables detection at nanogram levels. Lamivudine was used as the internal standard. The proposed method has been validated with a linear range of 10-600 ng/ml for TEN and 25-2,500 ng/ml for EMT. The intrarun and interrun precision values are within 12.0% for TEN and 15.6% for EMT at their respective LOQ levels. The overall recoveries for TEN and EMT were 84.3% and 68.5%, respectively. Total elution time was as low as 2 min.     

液相色谱串联质谱法测定人血清中齐多夫定的浓度

目的 建立液相色谱串联质谱法测定人血清中齐多夫定浓度.方法 血清样品用甲醇沉淀蛋白,内标为格列吡嗪,色谱柱为Lichrospher C18(4.6 mm×150 mm,5 μm),柱温为30℃,流动相为甲醇-10 mmol·L-1 NH4Ac(90:10,v/v),流速为0.5 ml·min-1.质谱采用ESI离子源负...     

Liquid chromatography-electrospray lonization tandem mass spectrometric determination of lornoxicam in human plasma

A rapid, sensitive and selective liquid chromatography-electrospray ionization tandem mass spectrometric (LC-ESI-MS/MS) method for the determination of lornoxicam in human plasma was developed. Lornoxicam and isoxicam (internal standard) were extracted from human plasma with ethyl acetate at acidic pH and analyzed on a Sunfire C18 column with the mobile phase of methanol:ammonium formate (10 mM, pH 3.0) (70:30, v/v). The analyte was detected using a mass spectrometer, equipped with electrospray ion source. The instrument was set in the multiple-reaction-monitoring mode. The standard curve was linear (r = 0.9998) over the concentration range of 0.50-500 ng/mL. The coefficient of variation and relative error for intra- and inter-assay at four QC levels were 0.7 to 4.2% and -4.5 to 5.0%, respectively. The recoveries of lornoxicam and isoxicam were 87.8% and 66.5%, respectively. The lower limit of quantification for lornoxicam was 0.50 ng/mL using a 200 pL plasma sample. This method was successfully applied to a pharmacokinetic study of lornoxicam after oral administration of lornoxicam (8 mg) to humans.     

Development and validation of a liquid chromatography with tandem mass spectrometry (LC-MS/MS) method for the determination of sunitinib in rabbit plasma

A liquid chromatography with tandem mass spectrometry (LC-MS/MS) method has been developed and validated for the measurement of sunitinib in rabbit plasma. After protein precipitation with acetonitrile, samples were analyzed on a Zorbax Extend-C₁₈ column (150 mm×4.6 mm, 5μm). The mobile phase consisted of a mixture of acetonitrile and deionized water (containing 0.05% formic acid)at a ratio of 27:73 (v/v), and the flow rate was set at 0.8 mL/min.The column temperature was maintained at 30 ^oC. The LC eluate was detected by an electrospray ionization (ESI) source operated in the positive ion mode, and quantification was conducted using MRM of the transitions m/z 399.24→283.01 and m/z 415.19→178.00 for sunitinib and internal standard (IS, diltiazem hydrochloride), respectively. The calibration curve was linear in the range of 2–600 ng/mL. The lower limit of quantification was 2 ng/mL. The method also exhibited satisfactory results in terms of sensitivity, specificity, accuracy (with relative error ranging from –4.0% to 1.1%), precision (with intra- and inter-day relative standard deviations ranging from 2.8% to 9.5%),matrix effect, recovery as well as stability. Taken together, our newly developed method was reliable to monitor sunitinib concentrations in rabbit plasma.     

Liquid chromatography tandem mass spectrometry method for simultaneous determination of antidiabetic drugs metformin and glyburide in human plasma

A simple and rapid liquid chromatography/tandem mass spectrometry (LC-MS/MS) method has been developed and validated for the simultaneous quantitation of antidiabetic drugs metformin and glyburide in human plasma using glimepiride as internal standard (IS). After acidic acetonitrile-induced protein precipitation of the plasma samples, metformin, glyburide and IS were chromatographed on reverse phase C18 (50 mm x 4.6 mm i.d., 5 microm) analytical column. Quantitation was performed on a triple quadrupole mass spectrometer employing electrospray ionization technique and operating in multiple reaction monitoring (MRM) and positive ion mode. The total chromatographic run time was 3.5 min and calibration curves were linear over the concentration range of 20-2500 ng/ml for metformin and 5-500 ng/ml for glyburide. The method was validated for selectivity, sensitivity, recovery, linearity, accuracy and precision, dilution integrity and stability studies. The recoveries obtained for the analytes and IS (>or=69%) were consistent and reproducible. Inter-batch and intra-batch coefficient of variation across four validation runs (LLOQ, LQC, MQC and HQC) was less than 8%. The accuracy determined at these levels was within +/-8% in terms of relative error (RE). The method was applied to a bioequivalence study of 500 mg metformin and 5mg of glyburide tablet after oral administration to 28 healthy human subjects under condition of fasting.     

亲水作用色谱串联质谱法测定人血浆中的拉米夫定

目的:建立简便、快速的亲水作用色谱串联质谱法(HILIC-MS/MS)测定人血浆中的拉米夫定。方法:以13C1,15N2(拉米夫定为内标,血浆样品经乙腈沉淀蛋白后,采用Luna HILIC(100 mm×3.0 mm,3μm)柱分离。流动相为乙腈-5 mmol·L-1醋酸铵-甲酸(95∶5∶0.01,v/v/v),进样体积2μL,样品分析时间3 min。采用ESI源正离子模式、多反应监测(MRM),用于定量分析的离子反应分别为m/z230(112(拉米夫定)和m/z233(115(内标)。结果:测定人血浆中拉米夫定的线性范围为8.0～2000 ng·mL-1,定量下限为8.0 ng·mL-1,日内、日间精密度(RSD)均小于9.0%,准确度(RE)在-7.1%~2.7%之间。本法成功应用于健康受试者口服2种拉米夫定片后的生物等效性研究。结论:采用稳定同位素内标的HILIC-MS/MS法更为简便、快捷和准确,适用于人血浆样品中拉米夫定的测定。     

HPLC法测定阿巴卡韦双夫定片中的含量

摘 要 目的： 建立HPLC法测定阿巴卡韦双夫定片中阿巴卡韦、拉米夫定、齐多夫定的含量。方法: 采用Hypersil C₁₈(200mm×4.6mm,5μm)色谱柱,以甲醇 庚烷磺酸钠溶液（取庚烷磺酸钠2.02 g,加三乙胺5 ml,加水至700 ml,用磷酸调pH至3.9）（30∶70）为流动相,流速为1.0 ml·min^-1,检测波长270 nm。结果: 拉米夫定、齐多夫定、阿巴卡韦分别在0.08～1.60 μg (r=0.999 4),0.24～4.80 μg (r=0.999 9),0.24～4.80 μg (r=0.999 9)范围内线性关系良好, 平均回收率分别为99.6%（RSD=0.9%）,99.7%(RSD=0.9%),99.9%（RSD=1.0%）。结论: 本方法简便、可靠、准确,可用于同时测定三组分含量。     

液相色谱-串联质谱法测定头发中11种阿片类生物碱

建立头发中海洛因、吗啡、单乙酰吗啡等11种阿片类生物碱的液相色谱-串联质谱测定方法,并考察海洛因滥用者头发中阿片类组分的存在情况。头发经冷冻研磨后加入硼酸缓冲液超声30 min,用氯仿-异丙醇(9∶1)提取。用Allure PFP丙基柱,以乙腈-乙酸铵(0.1%甲酸)梯度洗脱分离,采用二级质谱多反应监测模式(MRM)检测11种阿片类生物碱。头发中海洛因、吗啡、单乙酰吗啡等11种阿片类生物碱在对应质量浓度范围内线性良好(r>0.996 0);检测限(LOD)均小于0.05 ng.mg-1;回收率范围为47.2%～110%;日内精密度和日间精密度均小于14%。21例海洛因滥用者头发中均检出了海洛因、单乙酰吗啡、吗啡、可待因、乙酰可待因、氢可酮等主要组分。所建方法灵敏度高、选择性好,适用于同时分析头发中海洛因等11种阿片类生物碱组分,可有效鉴别海洛因滥用与阿片类药物或食品的摄取。     

液相色谱-串联质谱法测定大鼠血浆中脱水穿心莲内酯琥珀酸半酯的含量（英文）

目的建立了测定大鼠血浆中脱水穿心莲内酯琥珀酸半酯（DAS）的液相色谱-串联质谱法。方法血浆样品经液-液萃取后,以甲醇-水（70∶30,V/V）为流动相,通过Restek PinnacleⅡC18柱分离,格列吡嗪为内标,选择负离子扫描方式,以多反应监测（MRM）方式进行检测。用于定量分析的离子反应分别为m/z 531→m/z 431（DAS）和m/z 444→m/z319（格列吡嗪,内标）。结果DAS血浆浓度测定方法的线性范围为5～2500 ng/ml,定量下限为5 ng/ml。日内、日间精密度（RSD）均小于9.51%,准确度（RE）在-0.18%~1.93%。样品提取回收率为79.73%~85.99%,每个样品的测试时间为3 min。应用此法测试了大鼠口服或静注穿琥宁（DAS的单钾盐）后DAS的血药浓度,计算出其绝对生物利用度为3.69%。结论本方法灵敏度高、专属性强,适合于DAS的临床前药动学研究。