采用正负离子切换模式的LC-ESI-MS/MS法测定大鼠灌胃给药后京尼平苷的血浆浓度(英文)

建立了一种快速、专属性强的液相色谱-串联质谱方法(LC-MS/MS)测定中药栀子的主要活性成分–京尼平苷的大鼠血药浓度去评价其临床前药动学特征。京尼平苷血浆样品经沉淀蛋白后,采用DiamonsilC18色谱柱进行分析,采用流动相10mM醋酸铵–甲醇(20:80,v/v),流速0.6mL/min。样品测定采用"Truncated"多反应检测模式,采用正离子m/z411→411测定京尼平苷,采用负离子m/z415→295测定内标葛根素。线性浓度范围为10.0–5000ng/mL,最低定量下限为10.0ng/mL,样品提取回收率范围为84.8%–90.5%。该确证方法成功应用于大鼠灌胃(给药剂量200mg/kg)给予京尼平苷后的药动学研究。     

HPLC-ESI-MS联用法测定大鼠血浆中KJY-01浓度及其药动学研究

目的 建立测定KJY-01大鼠血浆中药物浓度的高效液相色谱-电喷雾离子化-质谱(HPLC-ESI-MS)联用的分析方法,对其进行大鼠体内的药动学研究.方法 取大鼠血浆50μL,加入内标酮康唑,用甲醇提取后取上清液吹干,用80%甲醇100μL溶解,取2μL进行HPLC-MS测定.色谱柱为Agilent ZORBAX Eclipse plus C18(150mm×2.1mm,5μm),流动相为20mmol醋酸铵(甲酸调节PH为3.75):甲醇(含1‰的甲酸)=22:78,流速为0.25mL/min,采用选择离子检测(SIM)法检测SQ109(m/z=331.3),KJY-01(m/z=475.4),酮康唑(内标,m/z=531.2).结果 SQ109的血药浓度在10～5000ng/mL范围内线性关系良好,最低检测限为10ng/mL,以质控样品计算,在各浓度水平下,此法的回收率均大于80%,日间和日内精密度小于20%,符合生物样品分析要求.结论 该方法操作简便、快速、灵敏、专属性强,可用于KJY-01的大鼠体内大批量样品定量分析及早期药动学研究.     

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。     

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。     

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。     

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。     

HPLC-MS／MS测定血浆中丁丙诺啡的浓度

目的建立人血浆中丁丙诺啡浓度的HPLC-MS/ MS 测定法。方法以曲马多为内标,将待测血浆样品经氢氧化钠碱化后,加正己烷提取,上清液经Agilent Eclipse plus C18 （4. 6 mm ×150 mm,3. 5 μm）色谱柱分离,甲醇- 水（80∶20,v∶v,含0. 005 mol/ L 甲酸铵和0.25%甲酸）为流动相,流速0. 5 mL/ min,进行洗脱,采用电喷雾电离源（ESI）,以多反应监测（MRM）方式进行正离子检测。用于定量分析的离子分别为m/ z 468. 3→55. 2 和m/ z 264. 2→58. 1（内标）。结果血浆中丁丙诺啡的浓度与峰面积比在50-20 000 ng/ L 的范围内线性关系良好,定量下限为50 ng/ L;提取回收率大于67%,日内日间精密度小于11%。结论该法简单、灵敏、快速,分离效果良好,可用于人体血药浓度测定和药动学研究。     

液-质联用法同时测定人血浆伊潘立酮及其两个代谢产物浓度

目的:建立LC-MS/MS法同时测定血浆伊潘立酮(iloperidone,ILP)及其两个代谢产物羟基伊潘立酮(hydroxy iloperidone,P88)和伊潘立酮羧酸(iloperidone carboxylic acid,P95)的药浓度。方法:色谱柱为Agilent C8(100 mm×4.6 mm,3.5μm),流动相为乙腈-2 mmol·L-1醋酸铵水溶液(含1.5%甲酸)=28∶72,流速为600μL·min-1,采用ESI源正离子模式,多反应监测方式测定。ILP,P88,P95的监测离子对分别为:m/z 427.2→233.2,m/z 429.2→261.2,m/z 429.2→233.2,内标氘代伊潘立酮(iloperidone-D3,ILP-D3)的监测离子对为m/z 430.2→233.1。结果:血浆ILP,P88和P95浓度分别在0.01~15 ng·m L-1,0.01~15ng·m L-1和0.02~20 ng·m L-1范围内线性均良好,日间、日内精密度均在可接受范围内,提取回收率较高,均大于75%。结论:该方法准确、灵敏度高、重现性好,能够较好地应用于ILP及其两个代谢产物的人体药动学研究。     

LC-ESI-MS法测定人血浆中紫杉醇的浓度及其在药代动力学中的应用(英文)

建立了一种灵敏度高、特异性好的高效液相色谱-电喷雾质谱法 (LC-ESI-MS) 测定人血浆中紫杉醇浓度的方法。采用一步液液萃取法进行血浆样品预处理, 提取液为甲基叔丁基醚, 内标选用炔诺酮。色谱柱为Zorbax SB-C18 柱 (100 mm×2.1 mm, 3.5 μm, Agilent), 流动相为甲醇-0.2 mmol/L甲酸铵缓冲盐溶液 (包含0.1%甲酸), 采用梯度洗脱。选择离子监测 (SIM) 的目标离子为紫杉醇的[M+Na]+ m/z 876.5和内标的[M+H]+ m/z 299.4。方法学验证表明线性范围是1.0-400 ng/mL (r>0.998), 最低定量限为1.0 ng/mL, 方法的批内和批间精密度都小于9.0%, 准确度在6.8%以内。此方法已成功应用于紫杉醇脂质体注射液在患者体内的药动学研究。     

UHPLC-MS/MS同时测定人血浆中辛伐他汀及其代谢产物

目的采用UHPLC-MS/MS同时测定人血浆中辛伐他汀及其代谢产物辛伐他汀酸,并研究辛伐他汀、辛伐他汀酸在人体内的药动学特征。方法血浆样品以乙醚萃取,采用UHPLC-MS/MS进行分析。色谱柱:Agilent ZORBAX SB-C18(100mm×2.1mm,3.5μm);乙腈-1mmol·L-1醋酸铵(甲酸调pH 4.5)为流动相梯度洗脱,流速:0.2 m L·min-1。采用电喷雾离子源(ESI),以多反应监测方式(MRM)进行定量分析。辛伐他汀和内标洛伐他汀在正离子模式下定量分析,离子对分别为m/z 419.4→199.3和m/z 405.3→199.3;辛伐他汀酸和内标洛伐他汀酸在负离子模式下定量分析,离子对分别为m/z435.5→115.2和m/z 421.4→101.2。结果辛伐他汀和辛伐他汀酸的线性范围均为0.2~50 ng·m L-1(r>0.99),最低定量限均为0.2 ng·m L-1,日内和日间精密度(RSD)均≤11.10%,提取回收率均≥63.71%。结论该方法专属性强、灵敏度高、重现性好,适用于辛伐他汀的药动学研究。     

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。     

Rapid quantification of gabapentin in human plasma by liquid chromatography/tandem mass spectrometry

A simple, sensitive and rapid liquid chromatography/tandem mass spectrometry (LC-MS/MS) method was developed and validated for the quantification of gabapentin, a new antiepileptic drug, in human plasma using its structural analogue, 1,1-cyclohexane diacetic acid monoamide (CAM) as internal standard. The method involved a simple protein precipitation by means of acetonitrile followed by a rapid isocratic elution with 10mM ammonium formate buffer/acetonitrile (20/80, v/v, pH 3.0) on Waters Symmetry C(18 reversed phase chromatographic column and analyzed by mass spectrometry in the multiple reaction monitoring mode. The precursor to product ion transitions of m/z 172-->154 and m/z 200-->182 were used to measure the analyte and the IS, respectively. The assay exhibited a linear dynamic range of 40-10000 ng/mL for gabapentin in human plasma. The limit of detection and lower limit of quantification in human plasma were 10 and 40 ng/mL, respectively. Acceptable precision and accuracy were obtained for concentrations over the standard curve ranges. A run time of 2 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.     

Simultaneous determination of amoxicillin and ambroxol in human plasma by LC-MS/MS: validation and application to pharmacokinetic study

A rapid, simple and sensitive LC-MS/MS method was developed for simultaneous determination of amoxicillin and ambroxol in human plasma using clenbuterol as internal standard (IS). The plasma samples were subjected to a simple protein precipitation with methanol. Separation was achieved on a Lichrospher C(18) column (150 mm x 4.6mm ID, dp 5 microm) using methanol (containing 0.2% of formic acid) and water (containing 0.2% of formic acid) as a mobile phase by gradient elution at a flow rate of 1.0 mL/min. Detection was performed using electrospray ionization in positive ion multiple reaction monitoring (MRM) mode by monitoring the ion transitions from m/z 365.9-->348.9 (amoxicillin), m/z 378.9-->263.6 (ambroxol) and m/z 277.0-->203.0 (IS). Calibration curves were linear in the concentration range of 5-20,000 ng/mL for amoxicillin, and 1-200 ng/mL for ambroxol, with the intra- and inter-run precisions of <9% and the accuracies of 100+/-7%. The method has been validated and applied to pharmacokinetic studies of compound amoxicillin and ambroxol hydrochloride tablets in healthy Chinese volunteers.     

A novel LC-ESI-MS-MS method for sensitive quantification of colchicine in human plasma: application to two case reports

A novel method based upon liquid chromatography coupled to ion trap mass spectrometry (MS) detection with electrospray ionization interface has been developed for the identification and quantification of colchicine in plasma or whole blood. Colchicine was isolated from plasma using a liquid-liquid extraction with dichloromethane at pH 8.0 and embutramide as an internal standard, with satisfactory extraction recoveries. Solutes were separated on a 3-microm C18 Uptisphere (Interchim) column (150 x 2.0-mm i.d.) using acetonitrile/2 mM NH4COOH pH 3.8 buffer (50:50, v/v) as the mobile phase with a flow-rate of 200 microL/min. Data were collected either in full-scan MS mode at m/z 100-450 or in full-scan MS-MS mode, selecting the ion m/z 400.1 for colchicine and m/z 294.1 for embutramide. The most intense daughter ion of colchicine (m/z 358.1) and embutramide (m/z 207.9) were used for quantification. Retention times were 2.40 and 4.25 min for colchicine and embutramide, respectively. Calibration curves were linear in the 0.50-50 ng/mL range. The limits of detection and quantification were 0.05 ng/mL and 0.50 ng/mL, respectively. The intra- and interassay precisions were < 14%, and the intra- and interassay accuracies were in the 97-105.8% range at either 2 or 20 ng/mL. A fatal case of colchicine self-poisoning with a lethal blood concentration of 60 ng/mL and nonfatal case with a plasma sample collected very late (at least 36 h after the ingestion) are presented. The described method enables the unambiguous identification and quantification of colchicine with a very good sensitivity, using only 1 mL of sample.     

高效液相色谱-质谱/质谱联用法测定人血浆中莫沙必利

建立测定人血浆中莫沙必利的高效液相色谱-质谱/质谱联用法。取血浆样品经液-液萃取后，以乙腈为有机相，0.3%甲酸水溶液为水相，采用梯度洗脱的方式，用C₁₈柱分离，通过电喷雾离子化，以多反应监测(MRM)方式进行正离子检测。莫沙必利线性范围为0.17~68.00 ng·mL^-1，定量下限为0.17 ng·mL^-1，每个样品测试时间仅2.8 min，日内、日间精密度(RSD)均小于13%，准确度(RE)在±6.3%范围内。应用此法研究了20名志愿者单剂量口服枸橼酸莫沙必利片后的药代动力学特点。该方法、灵敏、准确、快速，适用于莫沙必利的药代动力学及生物等效性研究。     

LC-MS方法研究氢溴酸高乌甲素在小鼠体内的药代动力学(英文)

本文建立了一种快速、灵敏的LC-MS法用于检测小鼠血浆中的高乌甲素浓度。采用ESI源和多反应监测(MRM)的方式进行检测,所选用的高乌甲素和内标延胡索乙素的反应离子对分别为m/z 585→535和m/z 356→m/z 192。该方法在3.0～2 000.0 ng·mL-1浓度内线性关系良好,定量下限为3.0 ng·mL-1,日内和日间精密度(RSD)均小于9.9%,准确度(RE)在±4.8%之内。氢溴酸高乌甲素分别以1.0、2.0和4.0 mg·kg-1单剂量静脉注射给予小鼠后,t1/2分别为0.47、0.48和0.49 h,AUC0-t分别为55.5、110.5和402.9 ng·h·mL-1。实验结果表明,氢溴酸高乌甲素单剂量静脉注射给予小鼠后,在低剂量(1.0～2.0 mg·kg-1)范围内其药动学行为符合线性动力学特征,当给药剂量(2.0 mg·kg-1)增大至4.0 mg·kg-1时,AUC增加至约4倍,而Vz和CL却显著降低,呈现非线性动力学特征,可能与高浓度下药物血浆蛋白结合率的降低有关。     

A fast and sensitive liquid chromatographic-tandem mass spectrometric method for assay of lorazepam and application to pharmacokinetic analysis

A fast and sensitive method of coupled high-performance liquid chromatography-electrospray tandem mass spectrometry for the assay of lorazepam in human plasma was developed. Plasma samples were simply treated with acetonitrile to precipitate and remove proteins and the isolated supernatants were directly injected into the HPLC/MS/MS system. Chromatographic separation was performed on a Zorbax C(18) (100 x 2.1 mm I.D.) column with a 65:35 (v/v) mixed solution of acetonitrile and 10mM aqueous formic acid being used as mobile phase. With diazepam as an internal standard, quantification was performed by selected reaction ion monitoring of the transitions of m/z 321--> m/z 275 for lorazepam and m/z 285--> m/z 193 for the internal standard. The assay was validated in the concentration range of 0.71-71.3 ng/ml in human plasma. A detection limit of 0.10 ng/ml for lorazepam was achieved, and inter- and intra-run precisions of better than 4.4% (R.S.D.) were observed. The developed method has been successfully applied for pharmacokinetic study of the drug in man.     

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].     

Quantification of vincristine and its major metabolite in human plasma by high-performance liquid chromatography/tandem mass spectrometry

An analytical method using electrospray ionization and high-performance liquid chromatography/tandem mass spectrometry (LC/ESI-MS/MS) was developed to quantify vincristine and M1, the CYP3A-mediated metabolite of vincristine, in human plasma. Vinblastine (internal standard), vincristine, and M1 in plasma were extracted in methylene chloride after acidification with TCAA. The analytes were separated on an Inertsil ODS-3 C18 column (2.1 x 150 mm) with a 5-mum particle size using a gradient elution with a run time of 20 min. The initial mobile phase composition was 0.2% formic acid/water (80:20, v/v) with a final composition of 0.2% formic acid/water (20:80, v/v). Detection was accomplished with multiple reaction monitoring for vinblastine (m/z 406.3--> 271.7), vincristine (m/z 413.2--> 362.2), and M1 (m/z 397.3 --> 376.2). At three concentrations of vincristine and M1, the inter-day and intra-day accuracy and precision were within the acceptable limits for validation (106.8 +/- 9.6% for intra-day, n = 5 each concentration; 90.9 +/- 10.9% for inter-day, n = 4 each concentration). For both vincristine and M1, the concentration limits of quantification and detection were 12 pg/mL and 6 pg/mL, respectively. Stability studies indicated that 80% of M1 degraded in plasma after 15 hours at room temperature (n = 3, high and low QC concentrations). Therefore, short plasma processing times (<30 min) are recommended. The assay was used successfully to quantify vincristine and M1 in pediatric plasma samples up to 24 hours after vincristine administration. Vincristine and M1 concentrations were within the limits of quantification for all patient plasma samples.     

高灵敏度LC-MS/MS法测定犬血浆中的坦洛新

犬口服盐酸坦洛新控释片后血浆药物浓度C_max小于10 ng·mL^-1，需建立测定犬血浆中坦洛新的高灵敏度液相色谱-串联质谱法(LC-MS/MS)。血浆样品加入内标苯海拉明，用正己烷-二氯甲烷(2∶1)萃取后，反相C₁₈色谱柱分离，以甲醇-乙腈-甲酸铵(30∶40∶30，v/v/v)为流动相，流速为0.4 mL·min^-1。选用大气压化学离子化源(APCI)三重四极杆串联质谱仪，以选择反应监测方式进行检测，用于定量分析的离子反应分别为m/z 409→228(坦洛新)和m/z 256→167(苯海拉明)。坦洛新线性范围为0.02～50 ng·mL^-1，定量下限为0.02 ng·mL^-1。批内、批间精密度(RSD)均小于9.72%，准确度(RE)在-2.61%～8.82%。本方法灵敏度高，专属性强，用于犬口服盐酸坦洛新控释片后的药代动力学研究。