液质联用法测定裸鼠血浆中吉西他滨浓度

目的:建立高效灵敏的液质联用法(LC-MS/MS)测定裸鼠血浆中吉西他滨浓度。方法:选用0.01%醋酸水和乙腈(80∶20)作为流动相;质谱检测选择正离子方式,多离子反应监测(MRM)扫描,用于吉西他滨和内标(头孢克洛)定量分析的离子反应对分别为m/z 264.1→m/z 112.0和m/z 368.1→m/z 174.1。结果:该方法线性范围为5～500 ng.mL-1,定量下限为5 ng.mL-1。日内日间精密度均<12.9%,准确度<3.4%。结论:本方法操作简便、快速,灵敏度高,能够满足裸鼠血浆中吉西他滨检测的要求。     

Bioanalytical method development and validation using incurred samples: Quantitative estimation of metformin in human K3EDTA plasma by LC–MS/MS

Bioanalytical methods should be reproducible and consistent when applying to clinical sample analysis. Incurred sample reanalyses (confirmatory reanalyses) are performed to support clinical data, and regulatory agencies evaluate the same before approval of bioequivalent products/drugs. A confirmatory reanalysis was demonstrated for metformin after administration of 2/500 mg repaglinide + metformin fixed dose formulation under fasted and fed conditions. The liquid chromatography tandem mass spectrometry (LC-MS/MS) method for determination of metformin in human plasma using metformin-d₆as an internal standard has been developed and validated. The ions transitions recorded in multiple reaction monitoring (MRM) were m/z 130.1→60.0 and 136.2→60.0 for metformin and metformin-d₆, respectively. The compounds were isolated by solid phase extraction and separated on a C12 reverse phase (Synergi MAX-RP 80A) column, using isocratic mobile phase flow at a flow rate of 0.8 mL/min. No matrix effect was observed within the linearity range of 10.2–1741.8 ng/mL (r² > 0.99). The acceptable result of confirmatory reanalysis further indicated stability of metformin in the presence of repaglinide. The assay method was found to be highly reproducible and was successfully applied for pharmacokinetic evaluations of metformin in fixed dose combinations with repaglinide.     

HPLC—MS／MS法测定人体血浆中格列美脲及其活性代谢物

目的：建立同时测定格列美脲（Glimepiride,G）及其活性代谢物羟基格列美脲（Hydroxyl—glimepiride,M1）血浆浓度的高效液相色谱一串联质谱（HPLC—MS／MS）法。方法：血浆样品酸化后,经乙酸乙酯萃取后浓集进样,色谱柱为PhenomenexGeminiC18（50mill×3．0mm,5μm）,甲醇：水：甲酸（80：20：0．1）为流动相,流速为0．2mL／min,柱温为室温,采用电喷雾（ESI）离子源,多反应离子检测模式,以格列齐特（Gliclazide,IS）为内标。G、M1和IS的检测离子对分别为质荷比（仇／z）491．4—352．4、507．4—352．4、324．4—127．2。结果：G和M1的线性范围分别为1．25～400ng／mL和0．313～100ng／mL,最低定量限分别为1．25和0．313ng／mL;批内批间精密度均小于10％,方法回收率均在96．4％～102．5％之间。结论：该方法简便快速、特异性高,可用于同时测定G和M．血药浓度及其人体药代动力学的研究。     

Determination of a dipeptidyl peptidase IV agonist, β-aminoacyl containing thiazolidine derivatives (KR-66223) in rat plasma by liquid chromatography-tandem mass spectrometry

A sensitive liquid chromatography/tandem mass spectrometry (LC-MS/MS) method was developed for a novel dipeptidyl peptidase IV agonist (DDP-IV) agonist, KR-66223, in rat plasma. It involves liquid-liquid extraction (LLE) followed by HPLC separation and electrospray ionization tandem mass spectrometry. KR-66223 and imipramine (IS) was separated on Gemini-NX C18 column with mixture of acetonitrile-ammonium formate (10mM) (90:10, v/v) as mobile phase. The ion transitions monitored were m/z 553.2→206.2 for KR-66223, m/z 281.3→86.1 for imipramine in multiple reaction monitoring (MRM) mode. The linear ranges of the assay were 0.003-10μg/ml with a correlation coefficient (R(2)) greater than 0.99 and the lower limit of quantification was 3ng/ml. The average recovery was 78.9% and 87.1% from rat plasma for KR-66223 and imipramine, respectively. The coefficients of variation of intra- and inter-assay were 3.9-14.4% and the relative error was 0.8-11.5%. The method was validated and successfully applied to the pharmacokinetic study of KR-66223 in rat.     

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.     

HPLC-MS/MS法测定人血浆中千金藤素的浓度

目的建立人血浆中千金藤素浓度的HPLC-MS/MS测定法。方法血浆样品1.0 mL经甲基叔丁基醚萃取后,以2.5 mmol/L醋酸铵(含0.05%甲酸)-甲醇(40∶60)为流动相,采用ZORBAX Eclipse XDB-C8(150 mm×4.6 mm,5μm)柱分离,采用电喷雾电离化(ESI)方式和多反应离子监测(MRM)模式进行正离子检测。用于定量分析的离子反应分别为m/z 607.2→365.1(千金藤素)和m/z 247.2→98.0(内标甲哌卡因)。结果千金藤素血药浓度线性范围为0.10～208.40 ng/mL,定量下限为0.10 ng/mL。低、中、高3个浓度提取回收率分别为72.3%、76.8%、79.5%。结论该法操作简便、快速、灵敏,适用于千金藤素在人体内的药代动力学研究。     

LC-MS／MS法测定全血中他克莫司浓度的方法学研究

目的：建立测定人全血中他克莫司液相色谱-串联质谱联用（LC-MS／MS）方法,并与微粒子免疫分析（MEIA）方法测定的结果进行了比较。方法：运用LC-MS／MS方法,血样经乙腈沉淀蛋白,上清液氮气吹干流动相复溶进样。色谱柱：Zorbax Extend—C18柱（150mm×4．6mm,5μm）;流动相：甲醇-乙腈-20mmol·L^-1醋酸胺溶液（80：15：5）;流速：0．4ml·min^-1。采用电喷雾离子化四极杆串联质谱,多反应检测方式测定样品的浓度。检测离子对分别为m／z821．7→m／z 768．4和m／z 821．7→m／z 786．5。结果：他克莫司在0．5～128ng·ml^-1浓度范围内,峰面积与浓度线性关系良好（r=0．9991,／2=5）,最低定量限为0．5ng·ml^-1,平均回收率为99．0％,日内精密度、日间精密度的RSD分别为3．38％和4．16％。MEIA分析采用标准IMX测定方法。两种方法比较,其标准差小于3．0,表明相关性好。结论：本方法具有良好的灵敏度、准确度、精确度及专属性,可用于他克莫司血药浓度监测和人体药药动力学研究。     

液相色谱-串联质谱法同时测定人血浆中二甲双胍和格列吡嗪

建立了快速、灵敏的液相色谱-串联质谱法测定人血浆中的二甲双胍和格列吡嗪。血浆样品经0.3%甲酸-乙腈(v/v)沉淀蛋白后，以乙腈-水-甲酸(70∶30∶0.3，v/v/v)为流动相，流速为0.50 mL·min^-1。Zorbax Extend C₁₈柱分离，采用大气压化学电离源；以选择反应监测(SRM)方式进行正离子检测。用于定量分析的离子反应分别为m/z 130→m/z 60(二甲双胍)，m/z 446→m/z 321(格列吡嗪)和m/z 256→m/z 167(内标，苯海拉明)。测定血浆中二甲双胍的线性范围为2.00～2 000 ng·mL^-1， 定量下限为2.00 ng·mL^-1； 格列吡嗪的线性范围为1.00～1 000 ng·mL^-1， 定量下限为1.00 ng·mL^-1。该方法专属性好，灵敏度高，准确快捷，适用于二甲双胍和格列吡嗪的临床药代动力学研究。     

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.     

大鼠血浆中奥昔布宁及其N-去乙基代谢物的UPLC-MS/MS法测定

建立了超高效液相色谱-串联质谱法同时测定大鼠血浆中的奥昔布宁及其主要代谢物N-去乙基奥昔布宁.用液-液萃取法处理血样,以盐酸奥昔布宁-D11为内标,采用ESI源正离子模式、多反应监测进行测定.检测离子对分别为m/z 358.2→124.1(奥昔布宁)、m/z 330.2→96.1 (N-去乙基奥昔布宁)和m/z 369.3→142.1(奥昔布宁-D11).奥昔布宁及N-去乙基奥昔布宁在0.5～100 ng/ml和0.2～40 ng/ml浓度范围内线性关系良好,日内、日间RSD均小于8％.     

Plasma pharmacokinetics of isorhapontigenin,a novel derivative of stilbenes,in mice by LC-MS/MS method

Isorhapontigenin (ISO), a novel derivative of stilbene compound, possessess good pharmacological activities such as antiviral, antioxidant, and anticancer. The purpose of this study is to investigate the pharmacokinetic of ISO in mice plasma, after oral administration of three doses (40, 80, and 160?mg/kg). Isorhapontigenin was detected by liquid chromatography–tandem mass spectrometry (LC-MS/MS), and a reliable LC-MS/MS assay for ISO in mice plasma was developed. The method was linear over a concentration range of 5–2000?ng/ml, and the lower limit of quantification was 5?ng/ml. The results indicated that there was a linear relationship between AUC_(0–t), AUC_(0–∞), and C_max of ISO and dosages. ISO underwent quick absorption and elimination in mice.     

Liquid chromatographic-tandem mass spectrometric method for the simultaneous quantitation of telmisartan and hydrochlorothiazide in human plasma

A rapid and sensitive method using liquid chromatography-tandem mass spectrometry (LC-MS/MS) has been developed for the simultaneous determination of telmisartan and hydrochlorothiazide in human plasma. Sample preparation involved liquid-liquid extraction with diethyl ether-dichloromethane (60:40, v/v). The analytes and internal standard, probenecid, were separated on a Venusil XBP-C(8) column using gradient elution with acetonitrile-10 mM ammonium acetate-formic acid at a flow rate of 1.2 mL/min. Detection was by electrospray negative ionization mass spectrometry using multiple reaction monitoring of the transitions at m/z 513.0-->469.4 for telmisartan, m/z 295.9-->268.9 for hydrochlorothiazide and m/z 283.9-->239.9 for probenecid. For both analytes, the method was linear in the range 1.00-600 ng/mL with intra- and inter-day precision (as relative standard deviation) 相似文献   

LC-MS/MS 法测定人血浆中多潘立酮的浓度

摘 要 目的:建立测定人血浆中多潘立酮浓度的高效液相-质谱联用色谱法。方法: 以普萘洛尔为内标,血浆样本经0.1 mol·Ll^-1氢氧化钠溶液处理后用乙醚萃取。采用XTerra MS C₁₈柱 （150 mm×2.1 mm,3.5 μm）为色谱柱,以乙腈-水-甲酸（80∶20,含0.3%的甲酸）为流动相,流速为0.2 ml·minl^-1。质谱条件为电喷雾离子源（ESI）,正离子方式多反应（MRM）扫描,离子选择通道分别为m/z 426.3 → m/z 175.1（多潘立酮）和m/z 260.3→ m/z 183.1（普萘洛尔）。结果:多潘立酮测定方法的线性范围为0.5～100 μg·L^-1,日内和日间精密度(RSD)均小于10 %。结论:本方法特异性强,灵敏度高,适用于临床试验中血浆样本的高通量分析。     

液相色谱-串联质谱法测定人血浆中盐酸舍曲林浓度

目的：建立测定人血浆中盐酸舍曲林浓度的液相色谱-串联质谱法。方法：以替米沙坦为内标,内标法定量。流动相：乙腈-10mmol·L^-1乙酸铵-1%甲酸（70：30：0.1）;质谱采用离子喷雾离子化源,扫描方式为多重反应监测（MRM）,用于定量分析的离子反应分别为m/z306.3→m/z159.1（舍曲林）和m/z515.2→m/z276.1（替米沙坦）。结果：舍曲林和替米沙坦的保留时间分别为2.22min和2.44min;舍曲林的线性范围为0.5～50.0ng·mL^-1,r=0.9992,回归方程：Y=0.0021＋0.0217X,最低检测浓度为0.5ng·mL^-1;提取回收率在82%～90%范围内;日内相对标准差〈4%,日间精密度〈13%。结论：此法适合人体血浆盐酸舍曲林浓度的监测及生物利用度研究,结果准确、可靠。     

Simultaneous Determination of Celecoxib, Erlotinib, and its Metabolite Desmethyl-Erlotinib (OSI-420) in Rat Plasma by Liquid chromatography/Tandem Mass Spectrometry with Positive/Negative Ion-Switching Electrospray Ionisation

A new method for the simultaneous determination of celecoxib, erlotinib, and its active metabolite desmethyl-erlotinib (OSI-420) in rat plasma, by liquid chromatography/tandem mass spectrometry with positive/negative ion-switching electrospray ionization mode, was developed and validated. Protein precipitation with methanol was selected as the method for preparing the samples. The analytes were separated on a reverse-phase C₁₈ column (50mm×4.6mm i.d., 3μ) using methanol: 2 mM ammonium acetate buffer, and pH 4.0 as the mobile phase at a flow rate 0.8 mL/min. Sitagliptin and Efervirenz were used as the internal standards for quantification. The determination was carried out on a Theremo Finnigan Quantam ultra triple-quadrupole mass spectrometer, operated in selected reaction monitoring (SRM) mode using the following transitions monitored simultaneously: positive m/z 394.5→278.1 for erlotinib, m/z 380.3→278.1 for desmethyl erlotinib (OSI-420), and negative m/z −380.1→ −316.3 for celecoxib. The limits of quantification (LOQs) were 1.5 ng/mL for Celecoxib, erlotinib, and OSI-420. Within- and between-day accuracy and precision of the validated method were within the acceptable limits of < 15% at all concentrations. The quantitation method was successfully applied for the simultaneous estimation of celecoxib, erlotinib, and desmethyl erlotinib in a pharmacokinetic study in Wistar rats.     

Simultaneous determination of clozapine and its N-desmethyl and N-oxide metabolites in plasma by liquid chromatography/electrospray tandem mass spectrometry and its application to plasma level monitoring in schizophrenic patients

A liquid chromatography tandem mass spectrometry (LC-MS-MS) assay method for the simultaneous determination of clozapine and its N-desmethyl (norclozapine) and N-oxide metabolites in human plasma is described. The compounds were extracted from plasma by a single step liquid-liquid extraction procedure and analyzed using a high performance liquid chromatography electrospray tandem mass spectrometer system. The compounds were eluted isocratically on a C-18 column, ionized using positive ion atmospheric pressure electrospray ionization method by a TurboIonspray source and analyzed using multiple reaction monitoring mode. The ion transitions monitored were m/z 327 --> m/z 270 for clozapine, m/z 313 --> m/z 192 for norclozapine, m/z 343 --> m/z 256 for clozapine-N-oxide and m/z 421--> m/z 201 for internal standard. The standard curves of clozapine, norclozapine and clozapine-N-oxide were linear over the range of 1 ng/ml to 1000 ng/ml when 0.5 ml of plasma was used for the analysis (r(2) >0.998). Three pooled plasma samples collected from patients who were treated with clozapine were used as long-term quality control samples to check the validity of spiked standard curve samples made at various times. The intra- and inter-assay variations for the spiked standard curve and quality control samples were less than 14%. These variations for the long-term patient quality control samples were less than 11%. The LC-MS-MS assay for simultaneous determination of clozapine, norclozapine and clozapine-N-oxide reported here is highly specific, sensitive, accurate and rapid. This method is currently being used for the plasma level monitoring of clozapine and its N-desmethyl and N-oxide metabolites in patients treated with clozapine. The plasma levels of clozapine, norclozapine and clozapine-N-oxide varied widely within and among patients. The data revealed that the norclozapine and clozapine N-oxide metabolites were present at about 58%+/-14% and 17%+/-6% of clozapine concentrations in plasma, respectively.     

HPLC—MS／MS测定人体血浆和尿液中帕洛诺司琼的浓度

目的建立一种简便、灵敏的测定人体血浆和尿液中帕洛诺司琼浓度的高效液相色谱一串联质谱（HPLC-MS／MS）方法。方法血浆、尿液样品分别采用甲醇沉淀处理后,选样分析。采用Agilent-ZORBAX-C18色谱柱（2．1mm×50mm,5fμm）．以乙腈-0．1％甲酸溶液为流动相,采用正离子,多反应监测方式测定样品浓度。用于定量分析的检测离子质荷电（m／z）297．2→m/z110．1（帕洛诺司琼）和DI／Z285．0→M／z193．0（内标）。结果帕洛诺司琼血浆样品在0．02～10ng·mL^-1与峰面积线性关系良好（r=0．9975）;定量下限（LLOQ）为0．02ng·L^-1;日内与日间RSD均〈10％;回收率在89．6％～114．0％。尿样在2．5～100ng·mL^-1与峰面积线性关系良好,7—0．9974;定量下限（LLOQ）为2．5ng·mL^-1;日内与日间RSD均〈10％;回收率在96．4％～113．4％。结论本方法简便快速、灵敏准确,适用于帕洛诺司琼在人体体内的药物动力学研究。     

LC-MS/MS法测定人血浆中加巴喷丁的浓度及其药动学研究

建立液相色谱串联质谱(LC-MS/MS)法测定人血浆中加巴喷丁的浓度并将其应用于人体药动学研究。取血浆样品经甲醇沉淀蛋白后,以甲醇0.2%甲酸水溶液(80∶20)为流动相,用Inertsil ODS-3 C18柱(50 mm×2.1 mm ID,3μm)分离,采用电喷雾离子源,以多反应监测(MRM)方式进行正离子检测,定量分析的离子反应分别为m/z 172→m/z 154(加巴喷丁)和m/z 130→m/z 71(内标二甲双胍)。加巴喷丁线性范围为40.8～8.16×103 ng.mL 1,定量限为40.8 ng.mL 1,每个样品测试时间仅2.2 min,日内、日间精密度(RSD)均小于12%,准确度(RE)在±6.4%范围内。应用此法研究了20名健康志愿者单剂量口服加巴喷丁胶囊600 mg后的药动学特点。该方法快速、专属、灵敏、适用性强,可应用于加巴喷丁的人体药动学研究。     

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.     

HPLC-MS／MS法测定大鼠血浆中兰索拉唑及其代谢产物的浓度

目的：建立大鼠血浆中兰索拉唑及其代谢产物5-羟基兰索拉唑、兰索拉唑砜的HPLC-MS／MS测定方法。方法：色谱条件：色谱柱：Diamonsil C18柱（150mm×2．1mm,5um）;流动相：乙腈-水（合0．01％甲酸及2mmol·L-1的醋酸铵（43：57,V／V）;流速：0．3ml·min-1;柱温：40℃;进样量10ul。质谱条件：电喷雾离子源（ESI）,以多反应监测离子方式测定兰索拉唑及其代谢产物,选择性监测质荷比（m／z）为368．0／163．9（兰索拉唑）,384．1／179．9（5-羟基兰索拉唑）,383．9／115．9（兰索拉唑砜）,326．0／280．1（内标奥美拉唑）。样品用乙腈沉淀蛋白处理。结果：兰索拉唑、5-羟基兰索拉唑、兰索拉唑砜的线性范围分别为11．40～4560．00,1．26～504．00,1．24～496．00ng·ml-1;定量下限分别为11．40,1．26,1．24ng·ml-1;批内、批间精密度RSD均〈15％。结论：该方法灵敏、准确、快速、专属性好,适用于兰索拉唑及其代谢产物在大鼠体内的药代动力学研究。