液-质联用法测定犬血浆中苯环喹溴铵的浓度

目的:建立HPLC-MS测定犬血浆中苯环喹溴铵浓度的方法。方法:色谱柱为Phenomenex C18(2)100A(Luna3μm,100mm×2.0mm),预柱为Luna C18(2)(3μm,10mm×2.0mm)柱,流动相为含1%甲酸的醋酸铵(8mmol.L-1)溶液-甲醇(47∶53),流速为0.20mL.min-1;ESI选择性正离子检测。结果:在5~3 600μg.L-1范围内,苯环喹溴铵血药浓度呈线性关系,r=0.999 4。最低检测限为3μg.L-1,日内、日间精密度RSD均小于9.3%。结论:本方法专属性强,灵敏度高,准确性好,适合狗血浆中苯环喹溴铵的药动学研究。     

LC-APCI-MS/MS法研究维氨酯在大鼠体内的排泄

目的：建立LC-APCI-MS/MS法测定大鼠胆汁、尿液和粪便中维氨酯的浓度,对该药在大鼠体内的排泄规律进行研究,为临床试验提供依据。方法：生物样本经处理后,采用AgilentTCC18色谱柱,甲醇-水-甲酸（9370.1）为流动相,辛伐他汀为内标,在三重四极杆串联质谱仪上,采用大气压化学离子化（APCI）、正离子方式选择性离子监测模式,维氨酯和内标的检测离子分别为m/z448/283和m/z419/285,分别测定尿液、胆汁和粪便中原型物的含量。结果：胆汁、尿样、粪便中维氨酯的回收率均大于89%,日间和日内的变异系数均小于10%。胆汁和尿液中维氨酯在0.0808～40.4ng/mL浓度范围内线性关系良好（r=0.9972和0.9949）,粪便中维氨酯在0.2525～101ng/mL浓度范围内线性关系良好（r=0.9990）,维氨酯的定量下限为0.2pg,方法符合生物样品分析要求。维氨酯在粪便中72h内、在尿液中96h内、在胆汁中24h内原型药的平均累积排泄量分别为12.9%、0.003%、0.0001%。结论：该方法灵敏度高,专属性强,样品前处理简便,分析速度快,可满足大鼠排泄研究中药物浓度测定的要求,大鼠灌胃给药（5mg/kg）后维氨酯从尿液和胆汁中排泄的原型药物很少。     

苯环喹溴铵长期经鼻给药对小鼠学习记忆能力的影响

目的观察苯环喹溴铵对小鼠学习记忆能力的影响及其形成原因。方法 50只筛选出的雄性昆明小鼠随机分为5组:溶媒组,苯环喹溴铵高、中、低剂量组(5.86、2.93、1.47mg·kg~(-1))及戊乙奎醚组(2.25mg·kg~(-1)),给药容量为5μL·10g~(-1),经鼻给药,每日1次,持续1mo。以Morris水迷宫和跳台实验测试苯环喹溴铵对其学习记忆能力的影响。另取50只昆明小鼠(雌雄各半)随机分为5组,以LC-ESIMS法测定小鼠在经鼻给予苯环喹溴铵5.86mg·k~g(-1)后10、45min及3、12、48h脑组织和血浆中苯环喹溴铵的浓度(给药后每个时间点各10只小鼠)。结果苯环喹溴铵各剂量组,对小鼠在Morris水迷宫和跳台实验中的学习记忆能力均无明显影响(P>0.05);小鼠在给药后各时间点,脑组织中苯环喹溴铵的浓度均在定量下限以下(苯环喹溴铵在脑匀浆中的定量下限为0.5405μg·L~(-1)),而给药后48h血浆中苯环喹溴铵的浓度为(4.01±0.52)μg·L~(-1)。结论苯环喹溴铵长期经鼻给药对小鼠的学习记忆能力没有影响,可能是由于其难以通过血脑屏障所致。     

SPE-HPLC法测定人血浆中喹那普利和代谢产物及其人体药代动力学研究

目的:建立 SPE—HPLC 法测定人血浆中喹那普利及其代谢产物喹那普利拉的浓度,以研究喹那普利及喹那普利拉在健康志愿者中的药代动力学和相对生物利用度。方法:应用 C_(18)固相萃取柱提取血浆中喹那普利及喹那普利拉,喹那普利色谱条件为:迪马 Diamonsil C_(18)柱(150 mm×4.6 mm,5μm);流动相为乙腈-0.1%醋酸溶液(40:60,v/v);流速1.0 mL·min~(-1)。喹那普利拉色谱条件为:Phenomenex C_(18)柱(150 mm×4.6 mm,5μm);流动相为乙腈-0.005 mol·L~(-1)十二烷基磺酸钠(磷酸调 pH 2.5)(40:60,v/v);流速1.0 mL·min~(-1)。人体药代动力学试验采用单剂双周期交叉设计方案,将18名志愿者随机分为两组,分别口服参比制剂喹那普利片和试验制剂喹那普利胶囊各40 mg,清洗期为1周。结果:喹那普利的线性范围为10～800ng·mL~(-1),r=0.9931,最低定量限为10 ng·mL~(-1);提取回收率与方法回收率分别为82.1%～94.4%,92.6%～99.9%;日内、日间 RSD 均小于10%。喹那普利拉的线性范围为20～1200 ng·mL~(-1),r=0.9995,最低定量限为20 ng·mL~(-1);提取回收率与方法回收率分别为73.3%～94.0%,100.0%～105.9%;日内、日间 RSD 均小于7%。结论:本方法灵敏度高、特异性强、重复性好,测定结果可靠,统计学分析表明2种制剂的主要药代动力学参数无显著性差异,为等效制剂。     

人血浆中齐拉西酮的LC-MS/MS法测定

建立了固相萃取LC-MS／MS法测定人血浆中齐拉西酮浓度。采用C18柱，流动相为甲醇-水-甲酸（60：40：0.02，含5mmol／L醋酸铵，pH5.0），电离模式为ESI电离，扫描类型正离子扫描。在0．5-400ng／ml范围内线性关系良好（r=0．9999），最低定量限为0．5ng/ml。平均回收率大于93％，日内及日间RSD均小于10％。     

反相高效液相色谱-荧光法测定人血浆中维库溴铵的浓度

目的:建立以反相高效液相色谱-荧光法测定人血浆中维库溴铵浓度的方法。方法:色谱柱为Shim-pakCLC-ODS,流动相为1,4—二氧六环-0.1mol/L磷酸二氢钠∶0.11mol/L庚烷磺酸钠缓冲液(20∶80),样品提取分离后用流动相溶解,在Ex/Em=380/452nm波长处检测。结果:维库溴铵检测浓度在20~200ng/ml范围内线性关系良好(r=0.9995),最低检测浓度为10ng/ml(S/N=10),绝对回收率在98.63%~99.81%之间,方法回收率在98.83%~102.13%之间,日内、日间变异系数均小于10%。结论:本方法简便、稳定、灵敏,可满足临床药动学研究的需要。     

苯环喹溴铵在大鼠肝微粒体中的代谢转化研究

目的:建立检测大鼠肝微粒体中苯环喹溴铵代谢产物的方法,验证其在大鼠体内的代谢途径。方法:采用大鼠肝微粒体体外温孵法,建立液相色谱-质谱法测定并分析肝微粒体中苯环喹溴铵及其代谢物。色谱及质谱条件如下:色谱柱为TSK-gelODS-80Ts,流动相为甲醇-40mmol·L-1的乙酸铵水溶液(含0.1%甲酸)梯度洗脱;正离子模式,扫描型离子检测。结果:在体外代谢系统中,根据质谱碎片信息检测出6个代谢产物,分别是苯环喹溴铵的二羟基、单羟基和氧化产物。结论:建立的液相色谱-质谱法能够准确灵敏地测定大鼠肝微粒体中苯环喹溴铵的代谢产物,验证了在大鼠体内苯环喹溴铵的代谢部位在环戊烷基上。     

RP-HPLC测定头孢克肟的含量

目的建立测定头孢克肟含量的反相高效液相色谱法。方法采用HypersilBDSC18（4．6mm×250mm,10μm）色谱柱,流动相为0．1mol·L^-1醋酸铵缓冲液（用氨试液调pH为7．0）-乙腈（96：4）,流速1．0mL．min^-1,检测波长254nm,柱温35℃。结果头孢克肟浓度在102．38～1433．45μg·mL^-1内,峰面积与浓度呈良好线性关系（r=0．9999）;头孢克肟的最低检测限为0．18ng,最低定量限为0．60ng,含量测定结果与按BP2008年版法定标准方法测定结果没有显著性差异。结论本法简便、专属性强,可用于头孢克肟含量的测定。     

高压液相色谱法测定莫雷西嗪在家兔的血药浓度及药代动力学

建立了反相高效液相色普法测定血浆中莫雷西嗪浓度。采用内标法。流动相为甲醇－水－０.0065mol.L~(-1)醋酸铵－冰乙酸（７５：２４：０.０１５,v/v/v/v）,二氯甲烷提取。血药浓度在５０～５０００ng.mL~(-1)范围内呈线性关系,相关系数0.9997,血浆最低检测浓度４ng.mL~(-1).。方法回收率９６～１０２％,日内、日间ＲＳＤ为０.7～3.2和２.3~4.0％。应用该法研究了兔静脉注射药动学,用二室模型拟合,消除相半衰期为6.35±2.23h。本法简便、回收率和灵敏度高、重复性好,分析周期短,适于临床药代动力学和药效学的研究。     

HPLC法测定布洛芬缓释胶囊有关物质

目的：建立高效液相色谱法测定布洛芬缓释胶囊中的有关物质2-（4-丁基苯基）丙酸和4-异丁基苯乙酮。方法：采用十八烷基硅烷键合硅胶为填充剂的色谱柱,流动相为磷酸-乙腈-水（0.5：340：600,待平衡后加水稀释至1000mL）,检测波长214nm,进样量为20μL,流速为2.0mL/min。结果：布洛芬缓释胶囊中有关物质2-（4-丁基苯基）丙酸在3.00～9.00μg/mL浓度范围内线性关系良好（γ=0.9992）,平均回收率为99.9%,RSD为0.82%。检测限为0.1307μg/mL。4-异丁基苯乙酮在1.19～7.14μg/mL浓度范围内线性关系良好（γ=0.9995）,平均回收率为100.2%,RSD为1.00%,检测限为0.1428μg/mL。结论：该方法操作简单、准确,重现性、分离效果好,灵敏度高,能较好的进行布洛芬缓释胶囊中有关物质2-（4-丁基苯基）丙酸和4-异丁基苯乙酮的测定。     

Determination of bencycloquidium bromide in human urine using weak cation-exchange solid-phase extraction and LC–ESI-MS: Method validation and application to kinetic study of urinary excretion

A sensitive LC–ESI-MS method has been developed and validated for the determination of bencycloquidium bromide (BCQB) in human urine samples. The method utilized a solid-phase extraction (SPE) procedure, choosing carboxy propyl phase (CBA) as the extracting sorbent for purification of BCQB, with better baseline and higher selectivity achieved. Sample preparation by this method yielded very good and consistent mean recovery of above 94.5%. Another major benefit of the present method was the high detectability, with a lower limit of quantification (LLOQ) of 0.02 ng/ml. The developed method was successfully applied to determine BCQB in human urine, and was proved to be suitable for use in Phase I clinical pharmacokinetic study of BCQB.     

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。     

Oral bioavailability and enterohepatic recirculation of otilonium bromide in rats

This study was conducted to examine the oral bioavailability and the possibility of enterohepatic recirculation of otilonium bromide in rats. A sensitive LC/MS/MS assay (LLOQ 0.5 ng/mL) was developed for the determination of otilonium and applied to i.v. and oral administration studies in bile duct cannulated (BDC) and non-BDC rats. After i.v. injection to BDC rats (1 mg/kg as otilonium), average t_1/2, CL, V_z and AUC were 7.9 ± 1.9 h, 8.7 ± 3.1 mL/min/kg, 5.7 ± 1.4 L/kg and 2,088 ± 676 ng·h/mL, respectively, and these values were comparable to those found in non-BDC rats. The percentages of i.v. dose excreted unchanged in bile and urine in BDC rats were 11.6 ± 3.0 and 3.1 ± 0.7%, respectively. Upon oral administration to non-BDC rats (20 mg/kg as otilonium), t_1/2, C_max, T_max and AUC were 6.4 ± 1.3 h, 182.8 ± 44.6 ng/mL, 1.9 ± 1.6 h and 579 ± 113 ng·h/mL, respectively. The absolute oral bioavailability was low (1.1%), while the drug was preferentially distributed to gastrointestinal tissues. A secondary peak was observed in the serum concentration-time profiles in non-BDC rats following both i.v. and oral administration, indicating that otilonium bromide was subject to enterohepatic recirculation.     

HPLC-UV method development and validation for 16-dehydropregnenolone, a novel oral hypolipidaemic agent, in rat biological matrices for application to pharmacokinetic studies

An accurate and precise HPLC assay has been developed and validated for determination of dehydropregnenolone (DHP) in rat plasma, bile, urine and feces. Separation was achieved using a C-18 reversed phase column with a mobile phase comprising of acetonitrile and deionized water (55:45% v/v) using a UV detector, set at a wavelength of 248 nm. The method, applicable to 200-microl plasma, bile and urine, involved double extraction of the samples with n-hexane. The sample clean up for feces involved single extraction of 50 mg of sample with 3 ml of acetonitrile. The method was sensitive with a limit of quantitation of 20 ng/ml in all the matrices and absolute recovery >92%. Precision and accuracy were within the acceptable limits, as indicated by relative standard deviation varying from 4.7 to 11.2% and bias values ranging from 1.8 to 8.8%. Moreover, DHP was stable in plasma, bile and urine up to 90 days of storage at -60 degrees C and after being subjected to three freeze-thaw cycles. The method was applied to generate the pharmacokinetics of DHP in rats after oral and intravenous administration.     

Discrimination of dimethylamphetamine and methamphetamine use: simultaneous determination of dimethylamphetamine-N-oxide and other metabolites in urine by high-performance liquid chromatography-electrospray ionization mass spectrometry

A simple and sensitive method by high-performance liquid chromatography-electrospray ionization mass spectrometry (LC-ESI-MS) has been investigated for the simultaneous determination of dimethylamphetamine (DMA), its specific yet labile main metabolite dimethylamphetamine-N-oxide (DMAO), and other metabolites, methamphetamine (MA) and amphetamine (AP), in urine. A combination of Bond Elut SCX columns for the solid-phase extraction of urine and a semi-micro SCX column for LC separations provided satisfactory results. The use of acetonitrile/5mM ammonium acetate buffer adjusted to pH 4 (65:35, v/v) as the mobile phase at a flow rate of 0.2 mL/min was found to be the most effective. The detection limits were 5 ng/mL for DMAO, 10 ng/mL for DMA and MA, and 50 ng/mL for AP in the SIM mode.     

Development and validation of an automated solid phase extraction and liquid chromatographic method for the determination of piperaquine in urine

A sensitive and specific bioanalytical method for determination of piperaquine in urine by automated solid-phase extraction (SPE) and liquid chromatography (LC) has been developed and validated. Buffered urine samples (containing internal standard) were loaded onto mixed phase (cation-exchange and octylsilica) SPE columns using an ASPEC XL SPE robot. Chromatographic separation was achieved on a Chromolith Performance RP-18e (100 mm x 4.6 mm I.D.) LC column with phosphate buffer (pH 2.5; 0.1 mol/L)-acetonitrile (92:8, v/v). Piperaquine was analysed at a flow rate of 3 mL/min with UV detection at 347 nm. A linear regression model on log-log transformed data was used for quantification. Within-day precision for piperaquine was 1.3% at 5000 ng/mL and 6.6% at 50 ng/mL. Between-day precision for piperaquine was 3.7% at 5000 ng/mL and 7.2% at 50 ng/mL. Total-assay precision for piperaquine over 4 days using five replicates each day (n = 20) was 4.0%, 5.2% and 9.8% at 5000, 500 and 50 ng/mL, respectively. The lower limit of quantification (LLOQ) was set to 3 ng/mL using 1 mL of urine, which could be lowered to 0.33 ng/mL when using 9 mL of urine and an increased injection volume.     

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.     

HPLC determination of polydatin in rat biological matrices: application to pharmacokinetic studies

A reversed-phase high-performance liquid chromatographic (RPHPLC) method has been developed for the determination of polydatin (PD) in rat plasma, bile, urine, feces and tissue homogenates using 2,3,5,4'-tetrahydroxychrysophenine-beta-d-glucoside as an internal standard. The sample pretreatment included deproteinization for plasma samples and a liquid-liquid extraction for bile, urine, feces and tissue homogenates. Separation was obtained on a C18 reversed-phase column with the mobile phase consisting of methanol and water (35:65 v/v). The flow rate was 1 ml/min and the effluent was monitored at 310 nm. The method showed good linearity over the concentration ranges employed for various matrices (r > 0.998). The quantification limits of PD in rat plasma, bile, urine, feces and tissue homogenates were 0.0251, 0.126, 0.025 microg/ml, 0.189 and 0.0378 microg/g, respectively. The accuracy and precision of the method were less than 12.0% for the various matrices. No interferences from endogenous substances were found. The method was successfully applied to study the pharmacokinetics of PD in rats after intravenous administration.     

左旋黄皮酰胺在大鼠体内的排泄

左旋黄皮酰胺[（-）-clausenamide]是从芸香科黄皮属植物黄皮[Clausena lansium（lour） sheels]叶的水浸膏中分离得到的有效成分，经不对称合成和拆分制备而得。药效学研究表明，左旋黄皮酰胺促进突触体谷氨酸释放，增加大鼠脑皮层厚度和海马CAL区突触数及NMDA受体密度，提高小鼠脑皮层和海马的胆碱乙酰转移酶活性，对抗樟柳碱引起的乙酰胆碱含量降低。这些结果表明左旋黄皮酰胺具有较好的促智和神经保护作用以及潜在的抗老年痴呆作用；右旋体作用不明显，且有较强的毒性。     

Determination of azelnidipine in human plasma by liquid chromatography-electrospray ionization-mass spectrometry

A liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) method for the determination of azelnidipine in human plasma was established. Nicardipine was used as the internal standard (IS). After adjustment to a basic pH with sodium hydroxide solution (0.1 M), plasma samples were extracted with cyclohexane-diethyl ether (1:1, v/v) and separated on a C(18) column with a mobile phase of 20 mM ammonium acetate solution-methanol-formic acid (25:75:0.5, v/v). The electrospray ionization was employed in a single quadrupole mass spectrometer for the determination. The method was linear over the concentration range of 0.05-40 ng/ml. The lower limit of quantification (LLOQ) was 0.05 ng/ml. The intra- and inter-run standard deviations were less than 9.5% and 11.0%, respectively. The method was successfully applied to study the pharmacokinetics of azelnidipine in healthy Chinese volunteers.