Determination of domperidone in human plasma using liquid chromatography coupled to tandem mass spectrometry and its pharmacokinetic study

A sensitive and selective liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the quantification of domperidone (CAS number: 57808-66-9) in human plasma using paracetamol (CAS number: 103-90-2) as an internal standard (IS). Domperidone and paracetamol in plasma were extracted with ethyl acetate, separated on a C18 reversed phase column, eluted with mobile phase of acetonitrile-glacial acetic acid (0.3%) (40:60, v/v), ionized by positive ion pneumatically assisted electrospray and detected in the multi-reaction monitoring mode using precursor→product ions of m/z 426.2→175.1 for domperidone and 152→110 for the IS, respectively. The calibration curve was linear (r2≥0.99, n=5) over the concentration range of 0.2-80 ng/mL and with lower limit of detection and quantitation of 0.05 and 0.2 ng/mL. The speci?city, matrix effect, recovery, sensitivity, linearity, accuracy, precision, and stabilities were validated for domperidone in human plasma. In conclusion, the validation results showed that this method was sensitive, economical and less toxic and it can successfully ful?ll the requirement of clinical pharmacokinetic study of domperidone oral preparation in Chinese healthy volunteers.     

LC-MS/MS法测定Wistar大鼠组织中银杏内酯B的浓度

目的建立LC-MS/MS法测定大鼠组织中银杏内酯B的浓度,并将此方法应用于银杏内酯B注射液在Wistar大鼠体内的组织分布研究。方法色谱柱:AQVASIL C18柱(100 mm×2.0 mm I.D.,5μm),流动相:乙腈-水-甲酸(体积比为60.0∶40.0∶0.4),流速:0.4 mL.min-1,离子源为电喷雾电离(electrospray ionization,ESI)源,负离子化方式,以选择反应监测(selective reaction monito-ring,SRM)方式进行扫描定量,用于定量分析的离子反应分别为m/z 423→m/z 367(银杏内酯B)和m/z 444→m/z 138(格列吡嗪)。测定了18只Wistar大鼠静脉注射给予银杏内酯B(给药剂量为3.75 mg.kg-1)后大鼠体内各组织和脏器中银杏内酯B的质量浓度。结果测定大鼠组织匀浆液中银杏内酯B的线性为1.0～500.0μg.L-1,定量下限为1.00μg.L-1。低、中、高3个质量浓度质控样品的日内和日间精密度分别小于6.4%和14.5%,准确度为97.3%～103.6%。银杏内酯B在心、肝、脾、肺、肾、胰、胃、小肠、脑、肌肉、脂肪、子宫、卵巢和睾丸14种组织中(除脑组织6 h时间点外)均被检出。银杏内酯B在大鼠各组织中分布广泛,自各组织中消除也较快。结论该方法适用于银杏内酯B注射液在Wistar大鼠体内的组织分布研究。     

人血浆中卡巴拉汀及其代谢物NAP226-90的LC-MS/MS测定研究

目的：建立LC-MS/MS法同时检测卡巴拉汀及其代谢物NAP226-90血药浓度。方法：血浆经甲基叔丁基醚-二氯甲烷提取预处理,用Phenomenex-curocil PFP（250 mm×4.6 mm,5μm）色谱柱,0.1%甲酸0.05%甲酸铵溶液-0.1%甲酸0.05%甲酸铵甲醇为流动相,梯度洗脱分离,ESI正离子化三重四极杆质谱MRM测定,检测反应离子对：卡巴拉汀m/z 251.0→206.0、NAP226-90 m/z 166.0→121.0、内标（美托洛尔）m/z 268.4→74.3。结果：卡巴拉汀及NAP226-90血药浓度在0.2～30 ng·mL-1范围内均线性关系良好,定量下限均为0.2 ng·mL-1,经方法学验证符合生物样品测定要求。结论：建立的LC-MS/MS方法可用于重酒石酸卡巴拉汀胶囊人体药动学研究。     

Determination of the plasma pharmacokinetic and tissue distributions of swertiamarin in rats by liquid chromatography with tandem mass spectrometry

An LC-MS/MS method was developed for the quantification of swertiamarin (CAS 17388-39-5) in rat plasma and tissues using gentiopicroside as the internal standard (IS). Swertiamarin and an IS were extracted from plasma and tissues by a simple solid-phase extraction (SPE) procedure. Separation was achieved on a Phenomenex kinetex-C18 column (100 mm×2.1 mm, 2.6 μm) with an isocratic mobile phase consisting of methanol and water (22:78, v/v) with 0.1% acetic acid at a flow rate of 0.2 mL/min. The analyte and IS were detected by negative ion electrospray ionisation in multiple-reaction monitoring mode while monitoring the transitions of m/z 433 [M + CH3COO] - →179 and m/z 415 [M + CH3COO] - →179 for swertiamarin and the IS, respectively. The method was validated with respect to selectivity, matrix effect, linearity, accuracy, precision, recovery and stability. The method was successfully applied in a pharmacokinetic study of swertiamarin after intravenous and oral administration to rats. The pharmacokinetics of swertiamarin showed rapid absorption and elimination, and its absolute bioavailability was low at 10.3%. After oral administration to rats, swertiamarin was rapidly and widely distributed in its tissues. High concentrations were found in the liver and kidney, indicating that swertiamarin was possibly absorbed in the liver and eliminated by the kidney.     

Rapid and simultaneous measurement of midazolam, 1'-hydroxymidazolam and digoxin by liquid chromatography/tandem mass spectrometry: application to an in vivo study to simultaneously measure P-glycoprotein and cytochrome P450 3A activity

In order to simultaneously determine in vivo P-glycoprotein (P-gp) and Cytochrome P450 3A (CYP3A) activity, a new, rapid and sensitive liquid chromatography/tandem mass spectrometry (LC-MS/MS) method has been developed and fully validated to simultaneously determine midazolam (MDZ, as CYP3A substrate), 1'-hydroxymidazolam (1'-OHMDZ) and digoxin (DG, as P-gp substrate) in rat plasma using digitoxin as the internal standard (IS). After a single step liquid-liquid extraction with tert-butyl methyl ether/dichloromethane (75:25, v/v), analytes were subjected to LC-MS/MS analysis using positive electro-spray ionization (ESI(+)) under selected reaction monitoring mode (SRM). Chromatographic separation was performed on an XTerra MS C18 column (50mm×2.1mm, i.d. 3.5μm). The MS/MS detection was conducted by monitoring the fragmentation of 326.05 → 244.00 (m/z) for MDZ, 342.02 →168.01 (m/z) for 1'-OHMDZ, 798.33 → 651.36(m/z) for DG and 782.67 → 635.24 (m/z) for IS. The method had a chromatographic running time of 3min and linear calibration curves over the concentrations of 2-400ng/mL for MDZ and 1'-OHMDZ and 0.5-100ng/mL for DG. The recoveries of the method were 86.8-96.3% for MDZ, 84.6-86.4% for 1'-OH MDZ, and 81.7-85.1% for DG. The lower limit of quantification (LLOQ) of the method was 2ng/mL for MDZ and 1'-OHMDZ and 0.5ng/mL for DG. The intra- and inter-batch precision were less than 15% for all quality control samples at concentrations of 5, 50 and 320ng/mL for MDZ and 1'-OHMDZ and 1, 10 and 80ng/mL for DG. The validated LC-MS/MS method has been successfully used to analyze the concentrations of MDZ, 1'-OH MDZ and DG in rat plasma for simultaneous measurement of in vivo P-gp and CYP 3A activity.     

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...     

Liquid chromatographic tandem mass spectrometry method for the quantification of miglitol in human plasma

A rapid, sensitive and accurate liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the quantification of miglitol (CAS 72432-03-2), an alpha-glucosidase inhibitor, in human plasma using gabapentin (CAS 60142-96-3) as internal standard (IS). Following protein precipitation, the analytes were separated using an isocratic mobile phase on a reversed phase phenyl column and analyzed by MS in the multiple reaction monitoring mode using the respective [M+H]+ ions, m/z 208/146 for miglitol and m/z 172/154 for the IS. The assay exhibited a linear dynamic range of 100-6000 ng/mL for miglitol in human plasma. The lower limit of quantification was 100 ng/mL with a relative standard deviation of less than 5 %. Acceptable precision and accuracy were obtained for concentrations over the standard curve ranges. The average absolute recoveries of miglitol and the IS from spiked plasma samples were 40.5 +/- 2.7 and 47.1 +/- 2.9 %, respectively. A run time of 2.5 min for each sample made it possible to analyze a throughput of more than 400 human plasma samples per day. The validated method has been successfully used to analyze human plasma samples for application in pharmacokinetic, bioavailability or bioequivalence studies. The miglitol plasma concentration profile could be obtained for pharmacokinetic study. The observed maximum plasma concentration (Cmax) of miglitol (100 mg oral dose) is 1740 ng/mL, time to observed maximum plasma concentration (tmax) is 3.5 h and elimination half-life (t(1/2)) is 2.5 h.     

Tissue distribution study of naringin in rats by liquid chromatography-tandem mass spectrometry

Naringin (CAS 10236-47-2), a flavanone glucoside widely present in fruits of citrus plants, has received extensive studies on its potential effects on health benefits and was recently demonstrated to be a putative antitussive. In this study, we determined the tissue distributions of naringin and its metabolites (naringenin and naringenin's conjugates) in rats to examine whether they undergo selective uptake by specific organs. Naringin was administered orally to rats at the dose of 42 mg/kg and the concentrations of naringin and its metabolites in tissue compartments were determined by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The areas under curve values in the individual tissues decreased as follows: stomach, small intestine, liver, trachea, muscle, kidney, lung, fat, heart, spleen, ovary, testis, brain for naringin; and liver, stomach, small intestine, kidney, trachea, lung, testis, heart, ovary, fat, spleen, muscle, brain for total naringenin (including free and its conjugates). Naringin and total naringenin were rapidly and widely distributed to all the tissues except brain in rats. They had difficulties in crossing the blood-brain barrier. There are no accumulations in rats. This study identifying naringin in several organs including lung and trachea may explain its effects as antitussive.     

超高效液相色谱-质谱法同时测定双黄连胶囊中的3种有效成分

目的 建立了同时测定双黄连胶囊中的绿原酸、黄芩苷和连翘苷3种有效成分的超高效液相色谱-电喷雾电离质谱(HPLC-ESI/MS)分析方法.方法 采用Waters BEH-C18色谱柱,以含0.2%甲酸的0.4mmol·L-1醋酸钠(A相)、甲醇(B相)为流动相进行梯度洗脱,在ESI正离子模式下,采用选择离子反应监测方法进行测定,用峰面积进行定量.结果 绿原酸、黄芩苷和连翘苷的线性范围分别0.05～50mg·L-1,0.10～500mg·L-1和0.01～25mg·L-1;检出限分别为0.010,0.020,0.002mg·L-1.3种成分的加样回收率为97.2%～101.6%,相对标准偏差小于2.3%.结论 该法快捷、准确、重复性好,可用于双黄连胶囊中的3种有效成分含量的同时测定.     

Quantitative determination of pidotimod in human plasma by liquid chromatography tandem mass spectrometry: application to a bioequivalence study

A highly sensitive and simple LC-MS/MS method after one-step protein precipitation was developed and validated for determination of pidotimod (CAS 121808-62-6) in human plasma using dextrophan (CAS 125-73-5) as internal standard (IS). Pidotimod and IS were separated on a YMC-ODS-AQ C18 column using 0.5% formic acid and methanol as a mobile phase at a flow rate of 0.3?mL/min. Detection was performed on positive ion mode of the transitions at 245.0→134.0 for pidotimod and 258.1→157.0 for IS by selected reaction monitoring (SRM). The assay exhibited a linear range of 0.05-10.0?μg/mL. The lower limit of quantification were 0.05?μg/mL. Validation results indicated that the accuracy as determined from quality control samples was in the range of -?4.00-6.48%. Intra-day and inter-day precision was ≤ 8.35% and ≤ 8.00%, respectively. The developed method was successfully applied to a bioequivalence study in 20 healthy Chinese volunteers following a single oral dose of 800?mg pidotimod. The simple, inexpensive protein precipitation and high-throughput method makes it a suitable and valuable tool in the investigation of the clinical pharmacokinetics and bioequivalence.     

LC-MS/MS法测定恒河猴血浆中巴替非班的浓度及药代动力学研究

目的:建立恒河猴血浆中巴替非班浓度的高效液相-串联质谱(Liquid Chromatography-TandemMass Spectrometry,LC-MS/MS)测定法,并研究其在恒河猴体内的药代动力学。方法:取恒河猴血浆200μL,加入含2μg.mL-1内标依替巴肽的乙腈-甲醇(70∶30)混合溶剂600μL沉淀蛋白,取上清液吹干,残留物加100μL流动相复溶,取上清液进行LC-MS/MS分析。色谱柱为ODS C18柱(150 mm×4.6 mm,5μm),流动相为甲醇-20 mmol.L-1甲酸铵(50∶50),流速为0.4 mL.min-1;质谱采用电喷雾离子化,正离子检测,巴替非班和内标的选择性检测离子分别为m/z 818.3→632.4和m/z 832.0→646.2。结果:巴替非班的线性范围为25～5 000 ng.mL-1,最低定量下限(lower limit of quantitation,LLOQ)为25 ng.mL-1,准确度、精密度及回收率均符合要求。结论:本方法专属性强,检测限低,灵敏度高,线性关系良好,方法简便快捷,适用于巴替非班临床前药代动力学研究。     

Ultra-performance liquid chromatography-tandem mass spectrometry method for the determination of nitrendipine in dog plasma and its application to a pharmacokinetic study of a solid self-emulsifying pellet formulation

A rapid, sensitive and selective ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed and validated for the quantitative determination of nitrendipine (NTD, CAS 39562-70-4) in dog plasma. Using propranolol hydrochloride (CAS 318-98-9) as an internal standard (IS), plasma samples pretreatment adopted a simple liquid-liquid extraction process with diethyl ether. Separation was carried out by a gradient elution on an Acquity UPLC BEH C18 column with a mobile phase consisting of water (containing 0.1% formic acid) and acetonitrile. Detection was performed by a triple-quadrupole mass spectrometry with positive electrospray ionization (ESI) as source ionization in multiple-reaction monitoring (MRM) mode at m/z 361.0 --> 315.0 for NTD and m/z 260.2 --> 116.0 for IS. The method demonstrated good linearity at the concentrations ranged from 0.1-200 ng/mL and the lower limit of quantification (LLOQ) of NTD was 0.1 ng/mL. The intra- and inter-day relative standard deviations (RSD) were less than 10%. The mean extraction recoveries of NTD and IS were 90.2% and 82.4%, respectively. Finally, the method was successfully applied to a pharmacokinetic study of home-made solid self-emulsifying pellets and conventional NTD tablets in beagle dogs following a single oral administration.     

UPLC-MS/MS法测定人血浆中卡马西平浓度

目的:建立UPLC-MS/MS法测定人血浆中卡马西平浓度并进行方法验证,应用于室间质评的样品测定。方法:采用UPLC-MS/MS法,以卡马西平-d8为内标,XSelect CSH C18柱(2.1 mm×75 mm,2.5μm)为分析柱,流动相A为含0.05%甲酸的水溶液,流动相B为含0.05%甲酸的乙腈溶液,流速为0.3 mL·min^-1,梯度洗脱;采用电喷雾离子源,以多反应监测(MRM)正离子模式进行检测,用于定量分析的离子对为m/z 237.1→194.1(卡马西平)、m/z 245.0→202.1(卡马西平-d8)。结果:卡马西平在5~1000 ng·mL^-1范围内线性关系良好(r≥0.9993),批内批间精密度和准确度、回收率和基质效应以及稳定性均符合生物样品测定要求。应用该方法测定的室间质评样品结果全部通过。结论:该方法准确、灵敏、选择性好、重现性高,适用于血浆中卡马西平浓度测定及室间质评。     

微透析联合LC-MS/MS法研究马钱子总碱囊泡凝胶透皮吸收

目的:建立皮肤微透析采样技术联合LC-MS/MS同时测定马钱子碱和士的宁的分析方法,并研究大鼠经皮给予马钱子囊泡凝胶后的透皮吸收。方法:应用皮肤微透析技术对大鼠给药部位皮下进行采样;微透析样品以他克林为内标,不经预处理直接进样;色谱采用XDB-C18柱,甲醇-乙腈-水(含0.05%甲酸和10 nmol·L-1甲酸铵)梯度洗脱;质谱采用正离子扫描多反应监测(MRM)方式。,用于定量的离子对分别为m/z 335.2→m/z 184.2(士的宁),m/z 395.2→m/z 324.2(马钱子碱),m/z 199.1→m/z 171.1(他克林)。结果:测定微透析样品中马钱子碱和士的宁2种成分的线性范围分别为0.195～50 ng.mL-1和0.156～40 ng.mL-1,日内和日间精密度(RSD)均小于15%,准确度、稳定性符合要求。结论:该方法操作简便、灵敏度高、专属性强,适用于皮肤微透析样品中马钱子碱和士的宁的测定。     

LC-MS/MS determination of the HIV-1 protease inhibitor indinavir in brain and testis of mice

A rapid and sensitive method for the determination of indinavir in mice brain and testis is described and validation data are provided. Indinavir and the internal standard (IS) amprenavir were isolated from homogenized tissue matrices using a mixed-mode solid-phase extraction (SPE) procedure and were then analyzed by reversed-phase liquid chromatography/tandem mass spectrometry (LC-MS/MS). The mass spectrometer in the positive-ion multiple reaction monitoring mode used pairs of ions at m/z of 614.1/421.3 for indinavir and of 506.1/245.3 for IS. The calibration curves were linear over the range 0.0012-0.0390 micromol/kg for brain and 0.39-12.50 micromol/kg for testis. Linearity, repeatability and accuracy were validated. The applicability of the method was demonstrated by assessing indinavir in brain and testis of three mice dosed with intravenous bolus administration of indinavir (16.3 micromol/kg).     

Quantification of azithromycin in human plasma by liquid chromatography tandem mass spectrometry: application to a bioequivalence study

A specific, sensitive and rapid liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) method has been developed and validated for the determination of azithromycin in human plasma. After deproteinizing the plasma sample with methanol, azithromycin and internal standard (IS: roxithromycin) were separated using a mobile phase comprised of acetonitrile : ammonium acetate buffer (50 mM, containing 0.05% acetic acid)=85:15 on a Hypersil GOLD C18 column (50 mm×2.1 mm ID, dp 1.9 μm). Detection was performed with a tandem mass spectrometer by selective reaction monitoring (SRM) through electrospray ionization. Target ions were monitored at [M+H]+ m/z 749.5→591.5 and 837.7→679.5 in positive electrospray ionization (ESI) mode for azithromycin and IS respectively. Linearity was established for the range of concentrations 2-800 ng/mL with a coefficient of correlation (r) of 0.9996. The lower limit of quantification (LLOQ) was identifiable and reproducible at 2.0 ng/mL. Both intra- and inter-batch standard deviations were less than 15%. The validated method was successfully applied to study the comparative bioavailability of azithromycin for suspension in test vs. reference in healthy Chinese volunteers through the statistical comparison of pharmacokinetic parameters obtained with the two formulations.     

Determination of propiverine hydrochloride in human plasma by high performance liquid chromatography-tandem mass spectrometry: application to the pharmacokinetic study of a sustained release formulation

A rapid, sensitive and reliable high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method was developed and validated for the determination of propiverine hydrochloride (CAS 54556-98-8) in human plasma using cetirizine di-hydrochloride as internal standard (IS, CAS 8388-51-0). Following liquid-liquid extraction with ethyl acetate, the separation was performed on a reverse phase C18 column with a mobile phase consisted of methanol-ammonium acetate (pH 4.0; 10 mM) (70:30, v/v). The detection was performed by a triple-quadrupole mass spectrometer in the positive ion and multiple reaction monitoring (MRM) mode, m/z 368.3 --> 116.1 for propiverine and m/z 389.2 --> 201.0 for the IS. The calibration curve fitted well over the concentration range of 0.2-200 ng/mL (all the concentration data in this study are related to salt (propiverine hydrochloride)). The limit of detection (LOD) and lower limit of quantification (LLOQ) in human plasma were 0.05 and 0.2 ng/mL, respectively. The method was proved to be rapid, sensitive, specific, accurate and reproducible and has been successfully applied to a pharmacokinetic study of propiverine hydrochloride sustained release capsules (the 30 mg dose in this study is related to 30 mg of salt (propiverine hydrochloride)). The major pharmacokinetic parameters in healthy Chinese volunteers are given for the first time and the sustained release characteristics of the sustained release formulation are evaluated. [corrected].     

色谱法测定人血浆中万古霉素的浓度

目的：建立HPLC法和液相色谱-串联质谱（LC-MS/MS）法检测人血浆中万古霉素的浓度,并分别与FPIA法进行比较。方法：HPLC法：100μL血浆样品加入等体积10%的高氯酸沉淀,振荡离心后取上清液进样。采用Agilent Eclipse XDB-C18（250 mm×4.6 mm,5μm）,柱温为35℃,流动相为0.05 mol·L-1 KH2PO4（pH=3.2）-甲醇（74∶26,v/v）,流速为1 mL·min-1,检测波长为230 nm。 LC-MS/MS法：100μL血浆样品用300μL乙腈沉淀,振荡离心后取上清液进样。色谱柱为Agilent Eclipse XDB-C18（100 mm ×2.1 mm,3.5μm）,柱温为40℃,流动相为水（含0.1%甲酸）-乙腈（90∶10,v/v）,流速为0.20 mL·min-1。采用电喷雾化离子源（ESI）,多离子反应模式（MRM）检测,万古霉素和内标去甲万古霉素的监测离子对分别为m/z 725.0→144.0和718.5→144.0。采用建立的HPLC法和LC-MS/MS法测定95份临床样本,并与FPIA法进行相关性和测定方法的偏倚分析。结果：HPLC法和LC-MS/MS法测定万古霉素的线性范围分别为1.2~96μg·mL-1和0.4~96μg·mL-1,低、中、高三种浓度质控品日内和日间相对标准差（RSD）均〈15%。两种方法与FPIA法有很强的相关性（r=0.9621,P〈0.0001和r=0.9466,P〈0.0001）,无明显偏倚。结论：建立的HPLC和LC-MS/MS法快速、灵敏、准确,样本测定结果与FPIA法无显著差异,适用于万古霉素的常规血药浓度监测及人体药物代谢动力学研究。     

HPLC-MS/MS同时测定人血浆中对乙酰氨基酚和咖啡因含量的方法学研究

目的建立同时测定人血浆中对乙酰氨基酚和咖啡因浓度的HPLC-MS/MS法。方法以茶碱为内标,血浆样品用甲醇沉淀蛋白后直接进样。用Waters symmetry C18（150 mm×4.6 mm,5μm）为分析柱,甲醇-0.2%醋酸=35∶65（v/v）为流动相,流速为0.9 mL·min^-1,采用柱后分流,0.2 mL·min^-1进入质谱,柱温35℃。选择监测的离子为m/z152.1→109.9（对乙酰氨基酚）、m/z195.3→138.1（咖啡因）和m/z181.1→123.9（茶碱）。结果血浆中对乙酰氨基酚和咖啡因的线性范围分别为0.02-4.04μg·mL^-1,5.05~1 010 ng·mL^-1;日内日间精密度RSD均〈7.94%。结论本方法专属性强,灵敏度高,操作简便、快速,符合生物样品分析要求,适用于临床药动学研究。     

LC-MS/MS法分析人血浆中西格列汀浓度及其应用研究

摘要：目的 建立一种测定人血浆中西格列汀浓度的液相色谱-串联质谱（LC-MS/MS）分析方法,并将该方法应 用于西格列汀在人体内的药代动力学研究。方法 以西格列汀-d4为内标,血浆样品经CleanertPPT沉淀板沉淀后, 通过Diamonsil C18色谱柱（100 mm×4.6 mm,5 μm）进行分离,使用甲醇-10 mmol/L甲酸铵水溶液（含10%甲醇,0.1% 甲酸）作为流动相,进行梯度洗脱,流速为0.5 mL/min。通过电喷雾电离源（ESI）,以多反应监测（MRM）模式进行正离 子检测。从选择性、残留、线性范围与定量下限、精密度与准确度、基质效应和回收率、稳定性方面进行方法学验证。 同时考察健康人口服西格列汀片100 mg后的主要药代动力学参数。结果 西格列汀、西格列汀-d4的MRM离子对 分别为 m/z 408.0→235.2、m/z 412.1→239.0。人血浆中西格列汀在 0.5～1 000 μg/L 浓度范围内线性关系良好（R2> 0.99）,定量下限为0.5 μg/L;定量下限和质控样品的批内、批间精密度（RSD）在0.83%~12.80%之间,准确度（RE）在± 10.0%以内。健康人口服西格列汀片100 mg后主要药代动力学参数：达峰时间（Tmax）、达峰浓度（Cmax）、、生物利用度 （AUC）、半衰期（T1/2）分别为（2.44±1.29）h、（375±138）μg/L、（2 915±585）h·μg/L、（11.10±2.41）h。结论 本LC-MS/ MS分析方法敏感度高且样品处理方法简单快速,满足生物分析的法规要求,可应用于人体内西格列汀的药代动力学 研究。