LC-MS/MS法测定比格犬血浆中匹莫范色林浓度及药代动力学研究

目的建立一种高效、灵敏的液相色谱-串联质谱(LC-MS/MS)法测定匹莫范色林在比格犬血浆中的浓度并应用于药代动力学研究。方法血浆样品用甲基叔丁基醚(MTBE)液液萃取预处理,使用Agilent Eclipse XDB-C 18(4.6 mm×150 mm,3.5μm)色谱柱进行分离,流动相为乙腈(含0.1%甲酸)-水(含5%乙腈,0.1%甲酸)梯度洗脱,流速为1.2 mL·min-1,分流比为1∶1,质谱检测用电喷雾离子(ESI)源,正离子模式下采用多反应监测(MRM)模式,分析时间为4.5 min。使用WinNonlin 6.1软件的NCA模块计算药动学参数。结果匹莫范色林在0.05~20 ng·mL-1浓度范围内线性关系良好,定量下限为0.05 ng·mL-1,批内和批间精密度良好CV均小于7.1%(LLOQ均小于18.4%),准确度RE均在-6.6%~12.7%之内(LLOQ均在3.8%~19.9%)。结论本方法高效、灵敏、准确,成功用于比格犬血浆中匹莫范色林浓度的测定及其药代动力学研究。     

LC-MS/MS法测定坦索罗辛在比格犬血浆中的浓度及其缓释制剂的药动学研究

目的:建立坦索罗辛血药浓度LC-MS/MS测定方法,并研究坦索罗辛缓释制剂在比格犬体内的药代动力学。方法:6只比格犬空腹单次口服给予含有0.4 mg的盐酸坦索罗辛缓释胶囊,给药前(0 h)、给药后不同时间点各采血0.3 mL,分离血浆后,加入内标愈创木酚甘油醚,经乙酸乙酯萃取,以甲醇:水:甲酸(80∶20∶0.2,V/V/V)为流动相,在反相C18色谱柱进行分离。采用电喷雾离子源(ESI源)的正离子方式检测,扫描方式为选择反应监测(SRM),用于定量分析的离子反应为m/z409.0→m/z228.0(坦索罗辛)和m/z199.0→m/z125.0(内标,愈创木酚甘油醚)。结果:坦索罗辛在线性范围内0.05～10.0 ng/mL线性良好,定量下限为0.05 ng/mL,批内、批间精密度(RSD)均小于13.4%。盐酸坦索罗辛缓释制剂0.4 mg口服给药后AUC0-24 h为(7.82±1.92)ng.h.mL-1、AUC0-∞为(8.04±2.02)ng.h.mL-1t、1/2为(2.96±2.13)h。结论:LC-MS/MS法灵敏度高、分析快速,适用于比格犬血浆中坦索罗辛的药动学研究,坦索罗辛缓释胶囊口服给药后半衰期短,消除快。     

LC-MS/MS测定大鼠血浆中乌头碱的浓度

目的:建立LC-MS/MS法检测大鼠血浆中乌头碱的浓度。方法:以维拉帕米作内标,血浆样品经MTBE液液萃取后,采用HyPURITYCyano(150mm×2.1mm,5μm)柱分离,流动相为乙腈:10mmol/L甲酸铵=(70∶30,V/V),流速为0.30mL/min。然后采用电喷雾离子源(ESI源)正离子多反应监测(MRM)扫描分析,乌头碱和维拉帕米的离子选择通道分别为:m/z646.4→586.4和455.2→164.9。结果:乌头碱的线性范围为9.3～2390pg/mL,最低检测浓度为9.3pg/mL,日内和日间变异均<15%。结论:本方法灵敏度高,适用于乌头碱在鼠内的药物代谢动力学研究。     

Determination of fexofenadine in human plasma by LC-MS/MS and its application in pharmacokinetic study

This study presented a simple, rapid, and sensitive liquid chromatography analytical method employing tandem mass spectrometry (LC-MS/MS) to determine fexofenadine in human plasma. After the de-proteination procedure with acetonitrile, chromatographic separation of fexofenadine was performed using a reversed-phase Eclipse XDB-C₈ column with a mobile phase consisted of 1 mmol/L ammonium acetate buffer solution containing 0.2% formic acid-methanol (45:55, v/v). Fexofenadine was quantified using tandem mass detection in the electrospray ionization (ESI) positive ion mode. The flow rate of the mobile phase was 1 mL/min, and the retention times of fexofenadine and the internal standard (IS, losartan) were 1.76 min and 2.65 min, respectively. The calibration curve was linear over the plasma concentration range of 1-1000 ng/mL. The relative standard deviations of intra- and inter-batches were less than 10.4% and 15.4%, respectively. The LC-MS/MS method reported in this study showed higher sensitivity for the quantification of fexofenadine in human plasma than that shown by previously described analytical methods. Lastly, the method was successfully applied to the pharmacokinetic of fexofenadine in healthy Taiwan volunteers.     

Determination of a novel derivative of the PAC-1 anticancer agent in rat plasma by LC-MS/MS and its application to a pharmacokinetics study

A new and sensitive liquid chromatography-tandem mass spectrometry method was developed for the determination of SM-1 in rat plasma. After a simple protein precipitation, SM-1 and internal standard (gefitinib) were separated with gradient elution on a Waters XBridge C₁₈ (50 mm×4.6 mm, 3.5 μm) using acetonitrile–methanol–10 mM ammonium acetate (37.5:37.5:25, v/v/v) as mobile phase. The triple quadruple mass spectrometer was set in positive electrospray ionization mode, multiple reaction monitoring was used for quantification. The precursors to produce ion transitions monitored for SM-1 and IS were m/z 407.3→203.4 and 447.3→128.3, respectively. The method validation was conducted over the curve range of 30–6000 ng/mL. The intra- and inter-day precisions were less than 4.7%, the average extraction recoveries ranged from 98.7% to 104.1% for each analyte. SM-1 was proved to be stable during sample storage preparation and analytical procedures. All the results met the acceptance criteria in accordance with the FDA guidance for bioanalytical method. Consequently, this method was successfully applied to determine SM-1 concentrations in rats after oral administrations at the doses of 200, 100 and even 50 mg/kg.     

液相色谱-串联质谱法测定人血浆中伪麻黄碱及其药动学研究

目的建立人血浆中伪麻黄碱含量的液相色谱-串联质谱（LC-MS/MS）测定法,并对其进行人体药动学研究。方法血样用甲醇沉淀、离心后进行质谱分析,色谱柱：Agilent ZORBAX SB C18（100 mm×3.5 mm,3.0μm）;流动相：甲醇-水（含10 mmol.L-1乙酸铵,0.05%甲酸）=28∶72（v/v）;质谱条件：气动辅助电喷雾离子化（ESI）,正离子检测,选择性多反应检测（SRM）：伪麻黄碱,166.1〉148.1;内标：帕罗西汀,330.1〉192.1。测定20名健康受试者口服受试制剂（单剂量、多剂量）后血浆中伪麻黄碱浓度。结果线性范围1-600μg.L-1,最低定量限（LLOQ）为1.0μg.L-1,绝对回收率为85.8%-87.6%。单剂量口服伪麻黄碱（120、240 mg）后药动学参数：t1/2为（6.8±0.9）、（6.6±1.3）h,tmax为（6.1±1.6）、（7.4±3.8）h,Cmax为（210.44±41.59）、（465.26±87.65）μg.L-1,V为（398.12±91.05）、（353.52±102.34）L,CL为（42.57±9.02）、（39.69±8.87）L.h-1,AUC0-48为（3 256.12±711.26）、（6 954.52±1 955.27）μg.h.L-1,MRT0-48为（11.71±0.89）、（11.87±1.26）h;伪麻黄碱多剂量（120 mg,bid）药动学参数：Cmax为（398.08±102.56）μg.L^-1,V为（309.66±82.37）L,CL为（37.51±7.23）L.h^-1,AUC0-48为（5 463.24±2 256.39）μg.h.L^-1,Cav为（282.11±92.19）μg.L-1,DF为（0.61±0.15）,AUCss为（3 372.85±1 328.78）μg.h.L^-1。结论本方法结果准确、灵敏,伪麻黄碱主要药动学参数与国内外文献报道基本一致。     

LC-MS/MS法测定人血浆中的金刚烷胺浓度及其药动学

目的建立HPLC-MS/MS分析方法测定人血浆中的金刚烷胺浓度,并用于研究健康受试者口服氨酚咖黄烷胺片后金刚烷胺的药动学。方法血浆经乙醚-二氯甲烷(4∶1,V/V)提取,采用C18柱为分析柱,甲醇-10 mmol.L-1醋酸铵溶液-冰醋酸(60∶40∶0.08,V/V/V)为流动相,流速1.0 mL.min-1,柱后分流,0.2 mL.min-1进入质谱,柱温35℃。质谱检测方式:多反应离子监测,选择监测的离子为m/z152→m/z135(金刚烷胺)和m/z275→m/z230(内标物,氯苯那敏)。结果金刚烷胺的线性范围为1~200μg.L-1(r=0.9999),血浆中金刚烷胺的最低定量限达1μg.L-1(RS,N>10),回收率>80%。结论本法简便、准确、灵敏度高,适用于金刚烷胺的药动学研究。     

LC—MS／MS测定人血浆中的罗红霉素

目的 采用液相色谱-质谱法测定人血浆中的罗红霉素.方法 采用API 3000型LC-MS/MS液质联用系统,Ulti-mate C<,18>分析柱(50 mm×4.6 mm,5 μm),流动相为甲醇-水(80∶20,含0.1%甲酸),流速0.3 ml·min<'-1>.MRM模式检测(离子对837.5/679.5),内标法定量.结果 罗红霉索和内标的保留时间分别为2.39、2.36 min,标准曲线0.01～10.00 μg·ml<'-1>的线性良好,最低定量限为10 ng·L<'-1>,日内、日间RSD分别小于3.4%、2.6%,方法 回收率98.8%～106.6%,萃取回收率70.7%～73.7%.结论 所建方法 快速简便、灵敏准确,适用于血浆中罗红霉素浓度的测定及药物动力学研究.     

LC-MS/MS法测定大鼠血浆中阿魏酸的血药浓度

目的建立高效液相色谱-串联质谱(LC-MS/MS)法测定大鼠血浆中阿魏酸的血药质量浓度。方法色谱柱采用Diamonsil C18柱(150 mm×4.6 mm,5μm),仪器采用API 4000三重四极杆串联质谱仪,采用多反应监测(multiple reaction monitoring,MRM)扫描方式检测,采用沉淀蛋白法处理样品,用于定量分析的离子对为m/z 193.1→m/z 134.1(阿魏酸)和m/z 121.2→m/z 77.0(苯甲酸,内标)。结果该方法的线性范围在5.0～2 000.0μg.L-1,定量下限为5.0μg.L-1,且血浆中内源性物质不干扰阿魏酸的测定;日内和日间精密度RSD均小于15%;阿魏酸的平均提取回收率为91.0%～100.5%,基质效应在87.8%～89.7%。结论该方法适用于阿魏酸在大鼠体内的药物动力学研究。     

LC-MS/MS法测定人血浆中盐酸舍曲林的浓度

目的建立LC-MS联用测定人血浆中盐酸舍曲林血药浓度的方法。方法采用Waters AlltimaTMC18柱(100mm×2．1 mm,3．0μm),柱温30℃;流动相:0．1％甲酸溶液-乙腈=40:60(V/V);流速0．2 mL·min-1;进样量2止;气动辅助电喷雾离子化(ESI),正离子检测,SRM,盐酸舍曲林:[M H]离子m/z 306．1,碎片子离子m/z 275．1,碰撞能量18 V;内标:盐酸帕罗西汀[M H]离子m/z 330．1,碎片子离子m/z 192．1,碰撞能量27 V。结果该法专属性好,对盐酸舍曲林的最低检测限为0．673μg·L-1,血浆样品检测的线性、精密度、回收率等指标均符合生物样品分析标准,在应用中取得良好的效果。结论本法可用于血浆中盐酸舍曲林的测定。     

Plasma pharmacokinetics and tissue distribution study of cajaninstilbene acid in rats by liquid chromatography with tandem mass spectrometry

Cajaninstilbene acid (CSA; 3-hydroxy-4-prenyl-5-methoxystilbene-2-carboxylic acid) is a major active constituent of pigeonpea leaves, has been proven to be effective in clinical treatment of diabetes, hepatitis, measles and dysentery. A rapid and sensitive liquid chromatography tandem mass spectrometry (LC–MS/MS) method was developed and validated for the determination of CSA in rat plasma and various tissues (brain, heart, lung, liver, spleen, small intestine and kidney) of rat for the first time. Rat plasma and tissue distribution pre-treated by protein precipitation with acetoacetate was analyzed using LC–MS/MS with an electrospray ionization (ESI) interface, and isoliquiritigenin was used as an internal standard. Chromatographic separation was achieved on a HIQ Sil C₁₈ column with the mobile phase of water and methanol (9:91, v/v) containing 0.1% formic acid and resulted in a total run time of 10 min. The isocratic elution mode pumped at a flow rate of 1.0 mL/min. The lower limit of quantification (LLOQ) which was 10 ng/mL. The calibration curve was linear from 10 to 6000 ng/mL (R = 0.9967) for plasma samples and 10 to 6000 ng/mL (R ≥ 0.9974) for tissue homogenates. The intra- and inter-day assay of precision in plasma and tissues ranged from 0.6% to 6.1% and 1.5% to 6.6%, respectively, and the intra- and inter-day assay accuracy was 93.5–104.6% and 93.3–107.5%, respectively. Recoveries in plasma and tissues ranged from 95.0% to 106.8%. The method was successfully applied in pharmacokinetic and tissue distribution studies of CSA after oral administration to rats. The pharmacokinetics of CSA showed rapid absorption and elimination (T_max, 10.7 ± 0.31 min; t_1/2, 51.40 ± 6.54 min). After oral administration in rats, CSA was rapidly and widely distributed in tissues. High concentrations were found in liver and kidney indicating that CSA was possibly absorbed by liver and eliminated by kidney.     

Determination of dabigatran in dog plasma by LC-MS/MS and its application to a pharmacokinetic study of dabigatran etexilate nanosuspension

In this study, a sensitive and rapid LC-MS/MS method was developed and validated to determine dabigatran in plasma of beagle dogs after oral administration of dabigatran etexilate nanosuspension (DABE-NS). The analytes (dabigatran) and sertraline hydrochloride (internal standard, IS) were separated on a Kromasil C₁₈ column using gradient elution consisting of methanol and formate buffer at a flow rate of 0.4 mL/min in 20 min. Detection and quantitation were carried out by multiple reaction monitoring following the transitions: m/z 472.17→289.07 and 305.98→275.00 for dabigatran and IS at positive ion mode, respectively. The calibration curves were linear from 1.0 to 500.0  ng/mL for dabigatran with r  = 0.9995. The accuracy of each analyte ranged from 94.8% to 107.1%, and the precision was within 6%. Besides, this method was successfully applied in the investigation of the pharmacokinetic profile of dabigatran in beagle dogs after oral administration ofDABE-NS. The maximum concentration and the areas under curves of dabigatranfor DABE-NS were significantly higher than those of control formulation, indicating improved oral absorption.     

LC-MS/MS测定人血浆中的辛伐他汀

目的建立测定人血浆中辛伐他汀的方法。方法采用LC-MS/MS法,血浆样品中加入内标洛伐他汀,经乙腈沉淀蛋白后,以正离子多反应监测(MRM)方式测试,用于定量的离子为m/z441.2→325.2(辛伐他汀)和m/z427.2→325.2(洛伐他汀)。色谱柱为Restek Allure C18(50 mm×2.1 mm,5μm),流动相为乙腈-0.5%甲酸水溶液(70:30)。结果血浆中辛伐他汀的线性范围为0.1-50.0 ng.ml-1,定量限为0.1 ng.ml-1,方法回收率为98%~102%,日内RSD<3.6%,日间RSD<5.6%。结论所建方法专属性好、灵敏度高,符合血浆样品测定的要求,适用于辛伐他汀的临床药物动力学研究。     

LC-MS/MS法测定伊曲康唑的血浆浓度及在生物等效性试验中的应用(英文)

目的建立液相色谱-串联质谱(LC MS/MS)法测定人血浆中伊曲康唑浓度方法,并评价伊曲康唑受试制剂和参比制剂是否具有生物等效性。方法采用蛋白沉淀的前处理方法,以10μL进样分析。色谱柱为CAPCELLPAKC18柱,流动相为V(乙腈)∶V(水)∶V(甲酸)=75∶25∶0.15),流速为0.20 mL.min-1。串联质谱API 3000采用正电喷雾离子源和多反应监测的扫描模式。结果伊曲康唑在质量浓度2～500μg.L-1内线性良好,定量下限为2μg.L-1,批内、批间的RSD均<10%。伊曲康唑受试制剂和参比制剂的相对生物利用度为(112.71±59.58)%,经方差分析及双单侧t检验表明两者具有生物等效性。结论该方法可以准确、快速和灵敏地应用于伊曲康唑的药动学与生物等效性研究,结果显示受试制剂和参比制剂生物等效。     

LC-MS/MS测定人血浆中的辛伐他汀

目的 建立测定人血浆中辛伐他汀的LC-MS/MS方法.方法 血浆样品经乙酸乙酯提取后,以乙腈-水(含0.1%乙酸和2.5 mmol·L-1甲酸胺)(90:10)为流动相,通过Agilent1100 VL型离子阱质谱仪,电喷雾离子源正离子模式,采用多反应离子监测方式测定辛伐他汀的浓度(MRM,m/z 441→325).结果 血浆中辛伐他汀的线性范围为0.5～10.0 ng·ml-1.定量下限为0.5 ng·ml-1.平均回收率85%～115%,日内、日间精密度在±15%之内.结论 所用方法高效、准确,特异性强,可用于辛伐他汀的药物动力学研究.     

LC-MS/MS法检测人血浆中甲基纳曲酮浓度的分析方法及其应用

目的:建立快速、灵敏、可靠的LC-MS/MS法测定人血浆中甲基纳曲酮的浓度.方法:采用弱阳离子交换固相萃取柱进行样品预处理,以溴甲基纳曲酮-d3为内标,通过Ultra PFP Propyl柱(100mm×2.1 mm,5.0 μm)色谱柱进行分离,流动相为20 mmol·L-1乙酸铵水溶液(含1％甲酸)和乙腈,等度洗脱...     

高效液相色谱-质谱联用(LC-MS/MS)法测定人血浆中多潘立酮的浓度

目的:建立高效液相色谱-质谱联用(LC-MS/MS)法测定人血浆中多潘立酮的浓度.方法:采用Waters公司SymmetryTM C18(3.9 mm×150 mm,5 μm)色谱柱;流动相为 20 mmol·L-1乙酸铵-甲醇(15∶85);流量为 0.6 mL·min-1;采用多反应离子监测(MRM)检测.结果:多潘立酮在0.3～50.0 ng·mL-1浓度范围内有良好的线性关系;最低检测浓度为 0.3 ng·mL-1;日内、日间RSD均小于15%;准确度均在±15%范围内.结论:本方法简单、快速、灵敏、准确,可用于人血浆中多潘立酮浓度的检测.     

LC-MS/MS法测定人血浆中的非索非那定及其药动学研究

目的:建立人血浆中非索非那定的LC-MS/MS测定方法,并研究其在健康人体内的药动学特征.方法:以格列吡嗪为内标,血样经甲醇沉淀法提取,采用Agilent ZORBAX SB-C18(100 mm×2.1 mm,3.5 μm),以甲醇-10 mmol·L-1醋酸铵水溶液(70∶ 30)为流动相,流速0.20 ml·min-1;质谱选择性检测非索非那定和内标格列吡嗪离子对分别为m/z 502.1 →466.2,446.0→321.1.20名健康受试者分成两组,分别进行低、中单剂量和多剂量给药的药物动力学试验.结果:非索非那定的线性范围为1.0 ～ 600.0 ng· ml-1(r=0.997 6,n=5),定量下限为1.0 ng·ml-1,提取回收率＞87.6％,日内和日间RSD≤8.5％.非索非那定的t1/2为(9.29～ 11.14)h,MRT为(7.95 ～8.24)h;多剂量给药后Css.max为(206.89±123.23)ng·ml-1,AUCss为(1122.98±646.60)ng·h·ml-1,DF为(1.81±0.36).结论:该法灵敏、简便、快捷、准确、重现性好,适用于非索非那定血药浓度测定及人体药动学研究.     

液-质联用法测定人体血浆中万古霉素的浓度

目的:运用液-质联用技术(LC-MS/MS),建立测定人体血浆中万古霉素浓度的方法,监测万古霉素血药浓度。方法:以替洛福韦为内标,乙腈沉淀蛋白后,经LC-MS/MS方法分析。采用色谱柱为安捷伦Poroshell 120 EC-C₁₈柱(4.6 mm×100 mm,2.7 μm),流动相为混合有机相(乙腈:甲醇-2:1)-0.1%甲酸水溶液(10:90);流速0.4 mL·min^-1;采用质谱电喷雾电离源(ESI),正离子模式,多重反应监测方式(MRM)检测;待测物质万古霉素用于定量的母子离子对:m/z 725.6→m/z 144.2,内标替洛福韦用于定量的母子离子对:m/z 288.0→m/z 176.2。结果:万古霉素在0.2~100 μg·mL^-1范围内线性关系良好,最低定量限为0.2 μg·mL^-1;日内RSD均在6%以下,日间RSD均在12%以下。提取回收率为58.01%~66.58%。结论:该分析方法快速、准确、灵敏度高、专属性强、重复性好,可用于医疗机构检测人体血浆中万古霉素的药物浓度。     

液相-质谱联用测定人血浆中尼群地平的浓度

目的:建立人血浆中尼群地平的HPLC-MS/MS定量分析方法。方法:血浆样品经液-液萃取预处理,色谱柱为Eclipse C18(4.6 mm×150 mm,5μm),流动相为乙腈-水梯度洗脱,采用三重四级杆质谱仪进行多反应监测方式(MRM),电喷雾离子源,对所建立方法进行了较全面的验证。结果:线性范围为0.1~50.1 ng/mL,最低定量浓度为0.1 ng/mL,提取回收率分别为84.1%、94.1%和87.0%,且稳定性良好。结论:该方法操作简便、快速、灵敏度高,可用来进行尼群地平的临床血药浓度测定和药动学研究。