LC/MS/MS法同时测定细胞孵化液中6种雌激素类化合物的含量

目的建立同时测定细胞孵化液中6种雌激素类化合物含量的LC/MS/MS法,并用该法研究E1S在乳腺癌细胞中的代谢行为。方法细胞孵化样品经处理后,采用Diamonsil C18柱分离,流动相为乙腈和5 mmol.L-1醋酸铵(pH 7.8),梯度洗脱,采用电喷雾电离(electrospray ionization,ESI)源,以多反应监测(multiple reaction monitoring,MRM)方式进行检测,用于定量分析的离子反应分别为m/z 269.3→m/z 145.2(雌酮,estrone,E1),m/z 271.1→m/z 145.2(雌二醇,estradiol,E2),m/z349.2→m/z 269.2(硫酸雌酮e,strone-3-sulfate,E1S),m/z 351.1→m/z 271.3(硫酸雌二醇,estradiol-3sulfate,E2S),m/z 445.5→m/z 269.3(β-D-葡萄糖醛酸雌酮,estrone-3-glucuronide,E1G),m/z 447.4→m/z 271.0(葡萄糖醛酸雌二醇,estradiol-3-glucuronide,E2G)和m/z 253.4→m/z 223.0(大豆苷元,内标)。结果该方法中E1、E2、E1S、E2S的线性分别为9.8～5 000.0 nmol.L-1,E1G、E2G的线性为0.98～500.00 nmol.L-1;日内和日间精密度均小于10.8%,相对误差在±11.7%以内。结论该法适用于细胞孵化液中6种雌激素类化合物的同时测定。     

LC-MS/MS方法测定大鼠血浆中亮丙瑞林浓度

目的建立迅速、简单、灵敏的液相色谱-串联质谱法测定大鼠血浆中亮丙瑞林的浓度。方法血浆样品采用固相萃取的方法,混合溶液(乙腈-水-甲酸=21.3:78.5:0.2,v/v/v)为流动相;采用Dikma C18柱(30 mm×4.6 mm,5μm)分离,通过电喷雾电离源(ESI),以选择性反应监测(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 605.5→221.1(亮丙瑞林)和m/z 584.5→221.1(内标,丙氨瑞林)。结果建立的人血浆内亮丙瑞林测定方法线性范围为0.02~100 ng·m L-1,定量下限可达0.02ng·m L-1。日内、日间精密度(RSD)均<13%。结论该方法预处理简洁,灵敏,专属性强,可方便用于大鼠血浆中亮丙瑞林的测定。     

LC/MS/MS法测定癫痫患儿血浆中拉莫三嗪的浓度

目的建立一种快速、灵敏地测定癫痫患儿血浆中拉莫三嗪(lamotrigine,LTG)的LC/MS/MS方法。方法采用高效液相色谱串联质谱电喷雾检测(LC/MS/MS)法,色谱柱为Welch Materials XB-C18柱,流动相甲醇-水(95∶5,V/V,含5 mmol/L甲酸铵),流速0.3ml/min,柱温40℃,以碱化的醋酸乙酯:二氯甲烷(4∶1)为提取剂。提取的样品经电喷雾离子源正离子化后,通过三重四级杆串联质谱仪,采用选择反应监测(SRM)对LTG(m/z 256.0→145.0)和内标阿普唑仑(m/z 309.3→281.1)进行测定。结果 LTG血药浓度在0.05~25μg/ml范围内线性关系良好(r=0.998 9),分析方法的最低检测限为5 ng/ml,其高、中、低(12.5、1.25、0.125μg/ml)3个浓度的平均提取回收率分别为82.97%、80.68%和83.33%,日内(n=5)、日间(n=15)RSD均<15%。结论本方法简单快捷、灵敏、准确、重现性好,可用于癫痫患儿LTG的临床血药浓度监测和药动学研究。     

HPLC-MS/MS法测定人尿液中戊乙奎醚的浓度

目的建立测定人尿液中戊乙奎醚浓度的HPLC-MS/MS法。方法尿样经稀释,离心后取上清液进行HPLC-MS/MS测定。色谱柱:Agilent ZORBAX Bonus-RP(4.6 mm×150 mm,5μm),流动相:5 m M醋酸铵(甲酸调pH 5.5)-甲醇(35∶65,v/v),流速:1.0 m L/min,柱温:30℃,进样量10μL。采用电喷雾离子源(ESI),正离子模式多重反应选择离子检测(MRM)。戊乙奎醚和内标的定量离子对分别为m/z 316.2→128.1和m/z 256.2→167.1。结果线性范围为1~100 ng/m L(r=0.997 7),最低定量限为1 ng/m L,日内和日间精密度(RSD)均<7.9%。结论本方法简单、快速、灵敏、专属性高,适用于人尿液中戊乙奎醚药动学的研究。     

HPLC-MS/MS法测定人血浆中甲氧氯普胺的浓度

目的建立测定人血浆中甲氧氯普胺浓度的LC-MS/MS方法。方法采用Alltech Alltima C18色谱柱(2.1 mm×50 mm,3μm),以5 mmol.L-1乙酸铵-乙腈-甲醇(体积比为50∶40∶10)为流动相,流速为0.2 mL.min-1,血浆样品在碱性条件下液-液萃取,通过电喷雾离子化四极杆串联质谱,以多离子反应监测(MRM)方式进行检测。用于定量分析的离子对分别为m/z300.20→m/z227.00(甲氧氯普胺)和m/z152.20→m/z134.00(内标,苯丙醇胺)。结果甲氧氯普胺线性范围为0.5~200.0μg.L-1,最低定量限为0.5μg.L-1,平均回收率为(71.38±6.35)%,日内和日间精密度均小于15%。结论该法适用于临床药物浓度监测和药物动力学的研究。     

HPLC-MS/MS法测定人体血浆中扎托布洛芬的浓度

目的:建立灵敏、快速和选择性高的高效液相色谱串联质谱法(HPLC-MS/MS)测定人血浆中扎托布洛芬浓度。方法:采用Agilent ZORBAX Eclipse Plus-C18(150 mm×4.6 mm,5μm)色谱柱,流动相为甲醇-0.1%甲酸水溶液(90:10,v/v),地西泮为内标,以多反应监测(MRM)扫描方式进行监测,监测离子质荷比:扎托布洛芬m/z 299.2→m/z 225.2,地西泮m/z 285.1→m/z193.1,血浆样品经乙腈沉淀蛋白后取上清液进样。结果:血浆中扎托布洛芬线性范围为0.02～20.0μg.mL-1,低、中、高3个浓度日内、日间精密度均小于10%。结论:本方法灵敏度高,选择性强,分析时间短,适用于人血浆中扎托布洛芬的浓度测定。     

UPLC-MS/MS法同时检测人血浆中拉莫三嗪和卡马西平浓度

目的采用高效液相色谱-质谱联用技术同时检测人血浆中拉莫三嗪和卡马西平浓度。方法采用UPLC-MS/MS法,以西替利嗪为内标,血浆经甲醇直接沉淀后进样分析。色谱柱为ACQUITY BEH C_(18)(2.1 mm×50 mm,1.7μm),流动相为甲醇-含0.1%甲酸的水溶液(70∶30,v/v),流速为0.2 ml/min,进样体积为10μl。质谱采用ESI离子源,正离子MRM模式扫描分析,拉莫三嗪、卡马西平和内标西替利嗪的离子对分别为m/z 256.0→211.2、m/z 237.0→194.1、m/z 389.2→201.2。结果拉莫三嗪和卡马西平线性范围分别为5～1 000 ng/ml (r=0.999 5)和5～1 000 ng/ml (r=0.997 5),定量下限均为5 ng/ml。批内、批间精密度RSD<15%;平均提取回收率分别为81.3%～84.9%和82.7%～87.3%。结论本方法样品前处理简便、快速,检测准确、灵敏度高,适用于临床治疗药物检测和药代动力学研究。     

LC-MS/MS法测定健康人血浆中的拉呋替丁

本文建立了一种快速测定人血浆中拉呋替丁血药浓度的LC-MS/MS法,采用CN柱(4.6 mm×150 mm,5μm),柱温30℃,流动相为甲醇-水(含20 mmol.L-1乙酸铵,0.2%甲酸)(97:3);流速1.0 mL.min-1;气动辅助电喷雾离子化(ESI),正离子检测,多反应离子检测(MRM)拉呋替丁和内标氯苯那敏分别为:m/z 432.20→351.15;m/z 275.10→230.05。拉呋替丁在1.98～396 ng.mL-1范围内线性关系良好(r=0.9991),定量限1.98 ng.mL-1。拉呋替丁低、中、高三个浓度批内及批间变异、准确度、绝对回收率均符合方法学要求,无显著基质效应。该方法专属性强,分析周期短,适合于临床上拉呋替丁血浆含量的测定。     

LC-MS/MS方法测定人血浆中多西环素浓度

目的建立迅速、简单、灵敏的液相色谱-串联质谱法测定人血浆中多西环素的浓度。方法血浆样品采用固相萃取的方法,以含0.04%TFA的混合溶液(甲醇-乙腈-水=45∶45∶10,v/v/v)为流动相;采用Dikma C18柱(30 mm×4.6 mm,5μm)分离,通过电喷雾电离源(ESI),以选择性反应监测(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 444.7→428.1(多西环素)和m/z 464.6→448.0(内标,去甲金霉素)。结果建立的人血浆内多西环素测定方法线性范围为50⁵ 000 ng·mL-1,定量下限可达50 ng·mL-1。日内、日间精密度(RSD)均<12%。结论该方法预处理简洁,灵敏,专属性强,可方便用于人血浆中多西环素的测定。     

LC-MS/MS内标法测定人血浆中西地那非的浓度

目的:建立LC-MS/MS法检测人血浆中西地那非的浓度。方法:以地西泮为内标,血浆样品经MTBE液液萃取后,采用Waters XTerraMS C18(150mm×2.1mm,5μm)柱分离,流动相为甲醇:10mmol/L醋酸铵=85∶15(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,西地那非和内标地西泮的离子选择通道分别为:m/z475.3→58.1和285.5→154.0。结果:西地那非的线性范围为1.50～765.90ng/mL,最低检测浓度为1.50ng/mL,日内和日间变异均<15%。结论:本法适用于西地那非的人体药物代谢动力学研究。     

Establishment of a liquid chromatographic/mass spectrometry method for quantification of tetrandrine in rat plasma and its application to pharmacokinetic study

A rapid and sensitive liquid chromatography-tandem mass spectrometric method (LC/MS/MS) for the determination of tetrandrine in rat plasma has been developed, fully validated and successfully applied to pharmacokinetic study in Sprague-Dawley (SD) rats after a single oral administration. Sample preparation involves a liquid-liquid extraction with n-hexane-dichlormethane (65:35, containing 1% 2-propanol isopropyl alcohol, v/v). Tetrandrine and brodimoprim (internal standard) were well separated by LC with a Dikma C(18) column using acetonitrile-methanol-ammonium formate aqueous solution (20mM) containing 0.3% formic acid (20:30:50, v/v/v) as mobile phase. Detection was performed on a triple quadrupole mass spectrometer in multiple reaction monitoring mode. The ionization was optimized using ESI(+) and selectivity was achieved using MS/MS analysis, m/z 623.0-->381.0 and m/z 339.0-->281.0 for tetrandrine and I.S., respectively. The present method exhibited good linearity over the concentration range of 5-2,000 ng/mL for tetrandrine in rat plasma with a lower limit of quantification of 5 ng/mL. The intra- and inter-day precision were 2.0-9.2% and 4.5-9.4%, and the intra- and inter-day accuracy ranged from -7.6 to 10.3% and -6.0 to 5.3%, respectively. No endogenous compounds were found to interfere with the analysis, and tetrandrine was stable during the whole assay period. The method was successfully applied to a pharmacokinetic study after an intragastric administration (i.g.) of tetrandrine to SD rats with a single dose of 50mg/kg. The results confirm that the assay is suitable for the pharmacokinetic study of tetrandrine.     

Simultaneous determination of the novel neuroprotective agent KR-31378 and its metabolite KR-31612 using high performance liquid chromatography with tandem mass spectrometry in human plasma

An LC/MS/MS method for the simultaneous determination of a neuroprotective agent for ischemia-reperfusion damage, KR-31378 and its N-acetyl metabolite KR-31612 in human plasma was developed. KR-31378, KR-31612 and the internal standard, KR-31543 were extracted from human plasma by liquid-liquid extraction. A reverse-phase HPLC separation was performed on Luna phenylhexyl column with the mixture of acetonitrile-5 mM ammonium formate (55:45, v/v) as mobile phase. The detection of analytes was performed using an electrospray ionization tandem mass spectrometry in the multiple reaction monitoring mode. The lower limits of quantification for KR-31378 and KR-31612 were 2.0 ng/ml. The method showed a satisfactory sensitivity, precision, accuracy, recovery and selectivity.     

A liquid chromatography/tandem mass spectrometry assay to quantitate MS-275 in human plasma

A rapid, sensitive and selective method was developed and validated using LC/MS/MS for determination of MS-275 in human plasma. Sample preparation involved a single step liquid-liquid extraction by the addition of 0.2 ml of plasma with 5 ml acetonitrile/n-butyl-chloride. Separation of the compounds of interest, including the internal standard paclitaxel, was achieved on a Waters X-Terra C(18) (50 mm x 2.1mm i.d., 3.5 microm) analytical column using a mobile phase consisting of acetonitrile/ammonium acetate (pH 2.9; 2mM)(60:40, v/v) containing 0.1% formic acid and isocratic flow at 0.15 ml/min for 3 min. The analytes were monitored by tandem-mass spectrometry with electrospray positive ionization. Linear calibration curves were generated over the range of 0.5-100 ng/ml with values for the coefficient of determination of >0.99. The values for both within day and between day precision and accuracy were well within the generally accepted criteria for analytical methods (<15%). This method was subsequently used to measure concentrations of MS-275 in cancer patients receiving an oral weekly dose of 4 mg/m(2).     

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.     

Simultaneous quantitation of hydrochlorothiazide and metoprolol in human plasma by liquid chromatography–tandem mass spectrometry

A rapid and sensitive method for the simultaneous quantitation of hydrochlorothiazide (HCT) and metoprolol (MET) in human plasma based on liquid chromatography–tandem mass spectrometry (LC–MS/MS) has been developed and validated. MS/MS detection involved switching the electrospray ionization (ESI) mode during chromatography from negative to detect HCT and its internal standard (I.S.) 5-bromouracil to positive to detect MET and its I.S. tramadol. Sample preparation by liquid–liquid extraction with diethyl ether–dichloromethane (60:40, v/v) was followed by chromatography on a Venusil MP-C18 column using methanol–ammonium acetate (10 mM)–formic acid (pH 3.4) (50:50:0.05, v/v/v) at a flow rate of 0.8 mL/min. The method was linear in the concentration range 3–1000 ng/mL for both HCT and MET using 100 μL human plasma. Intra- and inter-day precisions (as relative standard deviation) for HCT were 2.9–3.9% and 3.9–4.7%, respectively and for MET were 2.4–4.1% and 4.7–6.2%, respectively. Accuracies (as relative error) were ±3.8% and ±2.6% for HCT and MET, respectively. The assay was successfully applied to a pharmacokinetic study involving a single oral dose of a combination tablet (25 mg HCT, 50 mg MET) in healthy volunteers.     

Ultraperformance liquid chromatography tandem mass spectrometric method for direct quantification of salbutamol in urine samples in doping control

A fast and reliable quantitative method for salbutamol using direct analysis of the urine sample by ultraperformance liquid chromatography tandem mass spectrometry (UPLC/MS/MS) has been developed. Urine samples were spiked with salbutamol-d₆ (internal standard), and, then, they were diluted with ultrapure water (1:1, v/v). Aliquots of 1 μl of the mixture were directly analyzed by UPLC/MS/MS. The chromatographic separation was performed in a UPLC BEH C₁₈ (100 mm × 2.1 mm, 1.7 μm) column with a mobile phase contained 0.01% formic acid in ultrapure water (v/v) and 0.01% formic acid in acetonitrile (v/v), using gradient elution at 0.6 ml/min. The temperature of the column was set to 45 °C. The total run time was 3.2 min. Electrospray ionization in positive ion mode was used under multiple reaction monitoring (MRM) at different collision energies. Nitrogen and argon were used as desolvation and collision gas, respectively. The method was shown to be linear from 200 to 5000 ng/ml (r² > 0.99). The limit of quantitation was estimated in 200 ng/ml. Intra-assay precision and accuracies, evaluated by using quality control samples containing 550 and 1100 ng/ml salbutamol, were always better than 8.4%. The intermediate precision was estimated to be in the range of 5.6–8.9%. The method was shown to be reliable when applying to routine samples, and the short analysis time resulting from a simple sample preparation and a fast instrumental analysis makes it of great interest for antidoping control purposes.     

UPLC-MS/MS法检测复方芦丁片中芦丁的含量

目的建立超高效液相色谱-串联质谱法(UPLC-MS/MS)测定复方芦丁片中芦丁含量的方法。方法采用液相色谱串联质谱(LC-MS/MS)法。色谱柱为ACQUITY UPLC RBEH C 18色谱柱(2.1×100mm,1.7μm),流动相为甲醇-水溶液(42∶58),电喷雾离子化(ESI)正离子模式下选择质荷比(m/z)为301离子进行检测。结果所建方法能快速检测芦丁,在2.5~40.7ng·mL^-1浓度范围内线性关系良好(r=0.9995),平均加样回收率为99.5%,方法定量限为0.5ng·mL^-1,方法重复性和精密度良好。结论该方法专属性强,快速灵敏,适用于复方芦丁片中芦丁的含量测定。     

Liquid chromatography-tandem mass spectrometry for the determination of jaceosidin in rat plasma

Jaceosidin (4′,5,7-trihydroxy–3′,6-dimethoxyflavone), isolated from Artemisia species as well as Eupatorium species, has antiallergic, anticancer, anti-inflammatory and antioxidant activity. A rapid, sensitive and selective liquid chromatography-tandem mass spectrometric (LC/MS/MS) method for the quantification of jaceosidin in rat plasma was developed to characterize the pharmacokinetics of jaceosidin. Jaceosidin and the internal standard, linezolid, were extracted from rat plasma with ethyl acetate at acidic pH and analyzed on a Luna phenyl-hexyl column using the mixture of acetonitrile and 0.1% formic acid (45:55, v/v) as a mobile phase. The analytes were determined using an electrospray ionization tandem mass spectrometry in the multiple-reaction-monitoring mode. The calibration curve was linear (r² = 0.9973) over the concentration range of 2.00–500 ng/ml. The lower limit of quantification for jaceosidin was 2.0 ng/ml using 50 μl of plasma sample. The coefficients of variation of intra- and inter-assay at four QC levels were 2.4–9.6% and the relative errors were −9.1 to 10.0%. The matrix effects for jaceosidin and linezolid were practically absent. The recoveries of jaceosidin and linezolid were 87.0 and 87.7%, respectively. This method was successfully applied to the pharmacokinetic study of jaceosidin in rats.     

Validation of rapid and simple LC-MS/MS method for determination of voriconazole in rat plasma

A rapid, simple and sensitive LC-MS/MS analytical method was developed and validated for the determination of voriconazole (VRC) in rat plasma, using ketoconazole as internal standard (IS). Analysis was performed on a Shimadzu HPLC system using a Shimadzu C18 column and isocratic elution with acetonitrile-water-formic acid (60:40:0.05, v/v/v), at a flow of 1.0 mL/min (split ratio 1:5), and a mass spectrometer Micromass, equipped with a double quadrupole and an electrospray ionization interface, operated in a positive mode. Plasma samples were deproteinized with methanol (1:2) and 30 microL of the supernatant was injected into the system. The retention times of VRC and IS were approximately 3.3 and 2.7 min, respectively. Calibration curves in spiked plasma were linear over the concentration range of 50-2500 ng/mL with determination coefficient >0.98. The lower limit of quantification was 50 ng/mL. The accuracy of the method was within 5%. Intra- and inter-day relative standard deviations were less or equal to 12.5 and 7.7%, respectively. The applicability of the LC-MS-MS method for pharmacokinetic studies was tested using plasma samples obtained after intravenous administration of VRC to male Wistar rats. The reported method provided the necessary sensitivity, linearity, precision, accuracy, and specificity to allow the determination of VRC in pre-clinical pharmacokinetic studies.     

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 .