高效液相色谱-质谱联用法测定人血浆中氢吗啡酮的浓度

目的建立HPLC-MS/MS法测定人血浆中氢吗啡酮的质量浓度。方法用固相萃取法处理血浆,用Kromasil 100-5SIL和E97915色谱柱,以95%乙腈-5 mmol·L-1乙酸铵为流动相,梯度洗脱,用正离子、多反应监测方式测定样品质量浓度。结果血浆样品中,氢吗啡酮在0.05~10.00 ng·m L-1内线性关系良好(r=0.999 8),最低定量下限为0.05 ng·m L-1。血样日内与日间RSD均小于15%,平均回收率>70%,且稳定性均较好。结论本方法简便快速、灵敏准确、特异性强,适用于氢吗啡酮的体内药代动力学研究。     

高效液相色谱-质谱联用法测定人血浆中奥昔布宁的浓度

目的 建立一种用高效液相色谱-电喷雾离子化质谱联用技术测定奥昔布宁血药浓度的方法。方法 以0．0lmol／L乙酸铵水溶液-甲醇(15：85)为流动相，盐酸非洛普为内标，血浆样品经用环己烷萃取后上样，经C18柱分离后，以质谱为检测器，采用选择性离子检测(SIM)测定人体血浆中奥昔布宁的浓度。结果 线性范围0．2～50ng／mL(r=0．9948)，平均相对回收率在90％～110％之间，日内和日间精密度的RSD均小于10％，奥昔布宁的定量限为0．2ng／mL，提取回收率大于90％。结论 该方法快速、准确、灵敏，可用于奥昔布宁的药代动力学研究。     

超高效液相色谱-质谱联用法测定人血浆中依非韦伦的浓度

目的：建立超高效液相色谱-质谱联用法（UPLC-MS）定量测定人血浆中依非韦伦的浓度。方法：使用蛋白沉淀法处理血浆,以依非韦伦-D5为内标,色谱柱：Phenomenex Gemini C18（2 mm × 100 mm,5 μm,110 A）;柱温：40 ℃;流动相：乙腈-10 mmol·L-1乙酸铵（80∶20）,等度洗脱;流速：0.25 mL·min-1。负离子（ESI-）扫描模式,多反应监测方式（MRM）测定该药浓度。考察该方法的专属性、准确度与精密度、提取回收率与基质效应、全血稳定性及稀释可靠性等。结果：血浆样品中依非韦伦回归方程为y=0.494x+8.32×10-6（r = 0.999 1）,表明依非韦伦在0.1～8 μg·mL-1线性关系良好,定量下限为0.1 μg·mL-1。批内、批间精密度的RSD均＜15%,待测物及内标平均回收率＞95%且总体相等标准偏差（RSD）＜15%,稳定性良好。结论：该方法简便灵敏、准确可靠,适用于人体血浆中依非韦伦浓度的测定。     

液相色谱-质谱-质谱联用法测定人血浆中氯诺昔康液相色谱-质谱-质谱联用法测定人血浆中氯诺昔康

目的建立测定人血浆中氯诺昔康的液相色谱-质谱-质谱联用法,并用于中国受试者口服氯诺昔康的体内药代动力学研究。方法血浆样品经液-液萃取后,以甲醇-水-甲酸(80∶20∶0.5)为流动相,吡罗昔康为内标,采用Zorbax XDB-C8柱分离,通过液相色谱串联质谱,以选择反应监测(SRM)方式进行检测。定量分析离子反应分别为m/z 372→121(氯诺昔康)和m/z 332→121(吡罗昔康)。结果线性范围为2.0～1 600 μg·L^-1,定量下限为2.0 μg·L^-1。18名受试者po氯诺昔康8 mg后主要药动学参数t_1/2为(4.7±1.1) h,AUC_0-∞为(5.5±2.4) mg·h·L^-1。而另一名受试者t_1/2为105 h,AUC_0-∞为189.5 mg·h·L^-1。结论该法灵敏度高,线性范围宽,操作简便、快速,适用于临床药代动力学研究。     

高效液相色谱-质谱-质谱联用法测定人血浆中艾司西酞普兰浓度及其药代动力学研究

目的建立测定人血浆中艾司西酞普兰浓度的高效液相色谱-质谱-质谱联用法.方法血浆样品经甲醇沉淀后进行分析.色谱柱Lichrospher CN柱150 mm×4.6 m I.D.5μm,流动相甲醇水(含15 mmol·L-1乙酸铵)甲酸(72∶28O.1,v/v/v),流速1.0ml·min-1,电喷雾离子化三重四极杆串联质谱检测,以选择离子反应监测(SRM)扫描方式进行检测,采用选择离子反应监测(SRM)方式进行定量分析,用于监测的离子为m/z 325.0→234.0(西酞普兰)和m/z 409.1→238.1(氨氯地平,内标).结果线性范围为0.20～50.00ng·ml-1,最低定量浓度为0.20 ng·ml-1,应用此法测试了10名健康受试者口服草酸艾司西酞普兰片(10 mg)后不同时间的血药浓度,得到艾司西酞普兰药动学参数,Cmax为9.21±2.10 ng·ml-1,Tmax为3.75±1.04h,AUC0-t为514.6±152.3 ng·h·ml-1,AUC0-∞为540.5±162.3 ng·h·ml-1,t1/2为34.06±7.71 h及Ke为0.021±O.004h-1.结论该法专属、灵敏、简便、快速,适用于人血浆中艾司西酞普兰浓度的测定.     

高效液相色谱-质谱联用法测定人血浆中硝苯地平浓度

目的:建立高效液相色谱-质谱联用法测定人血浆中硝苯地平浓度。方法:以尼群地平为内标,血浆样品经乙腈沉淀后,经HPLC-MS/MS分离-分析。采用Diamonsil C18柱(2.1 mm×150 mm,5μm),以甲醇-乙腈-水(38.4∶38.4∶23.2)为流动相;流速:0.3 mL·min-1,采用电喷雾离子源(ESI),以多离子反应监测方式(MRM)进行正离子监测,硝苯地平和内标尼群地平的定量分析离子对分别为m/z369.1/224.2和m/z361.2/315.1。结果:硝苯地平血浆浓度测定方法线性范围为1.274～254.80 ng·mL-1,r=0.994 4。定量下限为1.274 ng·mL-1,方法回收率在85%～115%之间。日内和日间RSD小于11%。结论:本试验所建立起来的人血浆中硝苯地平浓度测定法灵敏、准确、可靠,适用于硝苯地平的人体内药动学研究。     

液相色谱-质谱联用法测定大鼠血浆中木通皂苷D的浓度

目的建立液相色谱-质谱联用法（LC-MS/MS）测定大鼠血浆中木通皂苷D的浓度。方法选用XDB-C18色谱柱,以5 mmol/L乙酸铵水溶液（含0.1%甲酸）-甲醇溶液为流动相,采用梯度洗脱进行分离。样本经甲醇沉淀后进样,选用3200QTRAP型质谱仪的多重反应监测扫描方式进行检测。结果木通皂苷D线性范围为10～1000 ng/ml,最低定量限为10ng/ml。准确度与精密度结果显示方法日间、日内变异均小于15%,相对误差为-2.8%～4.6%,低、中、高3个浓度提取回收率为95.3%～108.1%。结论本研究所建立的方法快速、灵敏、专属性强、重现性好,可用于大鼠血浆中木通皂苷D浓度的测定和药代动力学研究。     

液相色谱-质谱／质谱联用法测定人血浆中加兰他敏

目的:建立测定人血浆中加兰他敏浓度的液相色谱-质谱/质谱联用法,用于其人体血药浓度测定。方法:血浆样品经液-液萃取后,以甲醇-水-甲酸(65:35:1)为流动相,Zorbax SB-C_8柱(150mm×4.6mm,5μm)分离,采用大气压化学电离源,以选择反应监测方式进行正离子检测。内标为双氢吗啡酮,用于定量分析的离子反应分别为 m/z 288→m/z 213(加兰他敏)和 m/z 286→m/z 185(内标)。结果:血浆中加兰他敏最低定量限为0.5ng·mL~(-1),其线性范围为0.5～100ng·mL~(-1)。其高、中、低3个浓度的平均提取回收率为80.9%,方法回收率为100.5%,日内及日间 RSD 均<8%。结论:方法选择性强,灵敏度高,快速准确,可作为加兰他敏人体内药动学研究的手段。     

液相色谱-质谱-质谱联用法测定人血浆中阿德福韦的浓度

目的:建立快速、灵敏的液相色谱-质谱-质谱联用(LC-MS-MS)法测定人血浆中阿德福韦的浓度,并研究其在中国男性健康志愿者人体内的药动学。方法:以乙腈-0.1%甲酸为流动相,采用梯度洗脱,以5-溴尿嘧啶为内标。血浆样品经乙腈沉淀蛋白后用二氯甲烷提取,经ZorbaxXDB-C8柱分离后,通过电喷雾离子化三重四极杆串联质谱仪,以多反应检测(MRM)方式进行测定。结果:阿德福韦的线性范围为0.167～83.3μg.L-1(r=0.9989),平均相对回收率在88.3%～110.0%之间,日内、日间精密度的RSD均小于12%,阿德福韦的定量下限为0.167μg.L-1。结论:该方法具有快速、准确、灵敏等优点,适用于阿德福韦的药动学研究。     

高效液相色谱-串联质谱法测定人血浆中的缬沙坦

目的:建立灵敏、快速的高效液相色谱-串联质谱测定人血浆中缬沙坦浓度的方法,并用于人体药代动力学研究。方法:200μL的血浆样品经乙醚提取处理后,以乙腈-水-甲酸(70∶30∶0.2)为流动相,Diamonsil-C18柱分离,通过电喷雾离子源四极杆串联质谱,以多反应监测(MRM)的方式进行检测,选择监测离子反应为m/z436.3→291.2(缬沙坦)和423.1→207.1(氯沙坦)。结果:缬沙坦在2.120~5 300 ng.mL-1浓度范围内线性关系良好,定量限为2.120 ng.mL-1,日内、日间精密度(RSD)均≤9.0%,准确度在0.9%以内,缬沙坦和内标氯沙坦的回收率分别为83.1%和79.4%。结论:该法选择性强、灵敏度高,适用于缬沙坦的临床药动力学研究。     

高效液相色谱串联质谱联用法测定人血浆中克仑特罗的浓度

目的建立简单、快速和灵敏的高效液相色谱串联质谱联用法测定人血浆中克仑特罗的浓度。方法采用Agilent Eclipse XDB-C18色谱柱（4．6mm×50mm,1．8μm）,流动相为乙腈-0．1％甲酸=30：70（v/v）,流速为0．5mL·min^-1,柱温为25℃,通过电喷雾离子源进入质谱,以二级质谱多反应监测（MRM）方式对克仑特罗进行检测。结果克仑特罗的线性范围为5．4541090pg·mL^-1,LLOQ为5．45pg·mL^-1,平均相对回收率在80％～120％范围内,日内和日间变异均〈15％。结论本法简单、快速、灵敏、重现性好,能够用于克仑特罗的人体药物动力学和生物等效性研究。     

LC-MS/MS 法测定人血浆中倍他米松

建立测定人血浆中倍他米松的LC-MS/MS方法。采用Venusil XBP C₈ (200 mm×3.9 mm ID, 5 μm)色谱柱，流动相为甲醇-水(含甲酸铵5 mmol·L^-1)(80∶20)，流速0.4 mL·min^-1；质谱仪离子源为电喷雾离子源(ESI)，正离子模式检测，监测离子为393.3→355.2(倍他米松)和361.3→343.2(泼尼松龙,内标)。血浆样本用乙酸乙酯处理。倍他米松在0.5~80.0 ng·mL^-1线性关系良好(r=0.999 2)， 血浆低、 中、 高3种浓度(1.0， 10.0， 60.0 ng·mL^-1)平均提取回收率为88.24%，定量限为0.5 ng·mL^-1。本方法操作简便、准确、灵敏，适用于复方倍他米松注射液人体药代动力学研究。     

PMEA-Na在beagle犬血浆中的液相色谱/质谱/质谱联用法测定及其药代动力学

目的建立LC/MS/MS法测定犬血浆中PMEA-Na浓度,进行其药代动力学研究.方法血浆样品经甲醇沉淀蛋白后,采用多反应监测法测定其血药浓度.色谱柱为Xterra MS柱,流动相为甲醇水甲酸(25750.5),流速为 0.25 ml·min-1.Beagle犬分3个剂量组经静脉给药,给药剂量分别为 1.0、3.0 和 6.0 mg·kg-1.药代动力学参数通过DAS软件计算获得.结果PMEA-Na线性范围0.02～20 mg·L-1 (r=0.999);最低检测浓度为 20 μg·L-1,方法回收率为 97.1%～107.3%,日内日间变异分别小于 6.5%、10.8%.beagle 犬在 1.0, 3.0 与 6.0 mg·kg-1剂量下单次iv PMEA-Na后,测得其AUC分别为 2.3±0.5, 8.2±1.3 and 18.5±1.3 mg·L-1·h; t1/2 为 3.9±1.8, 8.4±1.5 and 8.9±0.6 h; CL为 0.44±0.09, 0.35±0.05 and 0.31±0.03 ml·h-1·kg-1.结论本方法专属性强,准确性好,可用于PMEA-Na血药浓度测定和药代动力学研究.     

HPLC-MS/MS法测定人血浆中雷喏嗪的质量浓度

目的建立HPLC-MS/MS法测定人血浆中雷喏嗪的质量浓度。方法以伏立康唑为内标,采用乙腈-体积分数0.1%甲酸水溶液(90∶10)为流动相,以Agilent-ZORBAX-C18柱(150 mm×2.1 mm,5.0μm)色谱柱为分析柱,通过电喷雾离子源(ESI),以正离子多反应监测(MRM)方式进行检测。雷喏嗪和伏立康唑用于定量分析的离子对分别为[M+H]+m/z428.5→m/z279.3和[M+H]+m/z350.2→m/z281.3。结果雷喏嗪线性范围为20~5 000 ng·mL-1,定量下限为20 ng·mL-1。日内和日间的RSD均小于15%,平均回收率大于75%。结论该方法专属性强,样品处理方便,灵敏度高,适用于雷喏嗪的临床药动学研究。     

Structure identification and elucidation of mosapride metabolites in human urine,feces and plasma by ultra performance liquid chromatography-tandem mass spectrometry method

Abstract

1.?Mosapride citrate (mosapride) is a potent gastroprokinetic agent. The only previous study on mosapride metabolism in human reported one phase I oxidative metabolite, des-p-fluorobenzyl mosapride, in human plasma and urine using HPLC method. Our aim was to identify mosapride phase I and phase II metabolites in human urine, feces and plasma using UPLC-ESI-MS/MS.

2.?A total of 16 metabolites were detected. To the best of our knowledge, 15 metabolites have not been reported previously in human.

3.?Two new metabolites, morpholine ring-opened mosapride (M15) and mosapride N-oxide (M16), alone with one known major metabolite, des-p-fluorobenzyl mosapride (M3), were identified by comparison with the reference standards prepared by our group. The chemical structures of seven phase I and six phase II metabolites of mosapride were elucidated based on UPLC–MS/MS analyses.

4.?There were two major phase I reactions, dealkylation and morpholine ring cleavage. Phase II reactions included glucuronide, glucose and sulfate conjugation. The comprehensive metabolic pathway of mosapride in human was proposed for the first time.

5.?The metabolites in humans were compared with those in rats reported previously. In addition to M10, the other 15 metabolites in humans were also found in rats. This result suggested that there was little qualitative species difference in the metabolism of mosapride between rats and humans.

6.?In all, 16 mosapride metabolites including 15 new metabolites were reported. These results allow a better understanding of mosapride disposition in human.     

Determination of febuxostat in human plasma using ultra‐performance liquid chromatography tandem mass spectrometry

A rapid and sensitive liquid chromatography‐tandem mass spectrometry method has been developed and validated for the determination of febuxostat in human plasma. The liquid‐liquid extraction technique was used for the extraction of febuxostat from human plasma using trandolapril as the internal standard (IS). Chromatography was performed on a ultra‐performance liquid chromatography (UPLC) BEH C18, 50 mm X 2.1 mm, 1.7 µm particle size column, with the mobile phase consisting of 0.1% formic acid and acetonitrile (in a 25:75 ratio), followed by detection using mass spectrometry. The method involves a simple reversed isocratic chromatography condition and mass spectrometry detection, which enables detection at sub‐microgram levels. The method was validated and the lower limit of quantification for febuxostat was found to be 0.075 µg/ml. The mean recovery for febuxostat ranged from 100.9 to 106.5%. This method increased the sensitivity and selectivity; resulting in high‐throughput analysis of febuxostat using commercially available IS for pharmacokinetic, bioavailability, and bioequivalence studies, with a chromatographic run time of 1.5 min only. Copyright © 2012 John Wiley & Sons, Ltd.     

Determination of olprinone in human plasma utilizing liquid chromatography tandem mass spectrometry

A sensitive and rapid method was developed for quantification of olprinone in human plasma utilizing liquid chromatography tandem mass spectrometry (LC–MS/MS). An aliquot of 1 mL plasma sample was extracted with ethyl acetate–dichloromethane. Separation of olprinone and the milrinone (internal standard, IS) from the interferences was achieved on a C₁₈ column followed by MS/MS detection. The analytes were monitored in the positive ionization mode. Multiple reaction monitoring using the transition of m/z 251 → m/z 155 and m/z 212 → m/z 140 was performed to quantify olprinone and IS, respectively. The method had a total chromatographic run time of 3 min and linear calibration curves over the concentration range of 0.5–60 ng/mL. The lower limit of quantification (LLOQ) was 0.5 ng/mL. The intra- and inter-day precisions were less than 16.3% for low QC level, and 7.1% for other QC levels, respectively. The intra- and inter-day relative errors were ranged between −12.2% and 3.7% for three QC concentration levels. The validated method was successfully applied to the quantification of olprinone concentration in human plasma after intravenous (i.v.) administration of olprinone at a constant rate of infusion of 2 μg/(kg min) for 5 min in order to evaluate the pharmacokinetics.     

液相色谱-串联质谱法测定人血浆中的利培酮

利培酮为苯并异噁唑类化合物,通过阻断5-HT2受体和多巴胺D2受体发挥抗精神病作用.同其他抗精神病药物相比,利培酮所引起的椎体外系副作用更少,药效更为明显 .     

Determination of dexmedetomidine in human plasma using high performance liquid chromatography coupled with tandem mass spectrometric detection: Application to a pharmacokinetic study

A rapid, sensitive and selective high performance liquid chromatography-electrospray ionization-tandem mass spectrometry method (HPLC-ESI-MS/MS) was developed and validated for the determination and pharmacokinetic investigation of dexmedetomidine (DMED) in human plasma. Dexmedetomidine and the internal standard (ondansetron) were extracted in a single step with diethyl-ether from 1.0 mL of alkalinized plasma. The mobile phase was a mixture of acetonitrile and 0.5% formic acid solution (30:70, v/v) at a flow rate of 0.2 mL min⁻¹. The detection was performed on a triple quadrupole tandem mass spectrometer in the selected reaction monitoring (SRM) mode using the respective [M+H]⁺ ions m/z 201.0 → 95.1 for DMED and m/z 294.1 → 170.1 for the IS. The assay exhibited a linear dynamic range of 5–5000 pg mL⁻¹ with the correlation coefficient above 0.9995. The lower limit of quantification (LLOQ) was 5 pg mL⁻¹ with a relative standard deviation of less than 15%. Acceptable precision and accuracy were obtained for concentrations over the standard curve range. The validated HPLC-MS/MS method has been successfully applied to study the pharmacokinetics of three level doses of DMED in Chinese healthy volunteers.     

液-质联用测定人血浆中罗红霉素浓度

目的:建立高效液相色谱-质谱联用法测定人血浆中罗红霉素浓度。方法:以克拉霉素为内标,血浆样品经乙腈沉淀后,经HPLC-MS/MS分离分析。采用Waters ODS C18柱(2.1mm×50mm,3.5μm),以甲醇-5mmol.L-1醋酸铵(含0.03%甲酸)(55∶45)为流动相;流速:0.2mL.min-1,采用电喷雾离子源(ESI),以多离子反应监测方式(MRM)进行正离子监测,罗红霉素和内标克拉霉素的定量分析离子对分别为m/z837.5→679.4和m/z748.5→590.3。结果:罗红霉素血浆浓度测定方法线性范围为0.02025~13.500mg.L-1,r=0.999 0。定量下限为0.02025mg.L-1,方法回收率在85%~115%之间。日内和日间RSD均小于15%。结论:本试验所建立的方法灵敏、准确、可靠,适于罗红霉素的人体内药动学研究。