复方氢溴酸东莨菪碱贴膏含量测定方法

目的：研究复方氢溴酸东莨菪碱贴膏中氢溴酸东莨菪碱含量测定方法.方法：采用加热回流提取复方氢溴酸东莨菪碱贴膏中的氢溴酸东莨菪碱,并用高效液相色谱法测定其含量.色谱柱：Agilent ZORBAX SB-C8柱（250 mm×4.6 mm,5 μm）,流动相：0.25%十二烷基硫酸钠溶液（用磷酸调节pH至2.5）-乙腈（60：40）,检测波长：210 nm,流速：1.0 ml·min^-1,进样量20 μl.结果：氢溴酸东莨菪碱浓度在11.24～224.80 μg·ml^-1范围内线性关系良好,r=0.999 9.平均加样回收率为98.22%,RSD为0.9%（n=6）.结论：所建含量测定方法简便准确,溶剂成本低,且较环保.     

阿霉素纳米胶束在大鼠体内的组织分布研究

目的建立大鼠组织样品的预处理及液相色谱-串联质谱(LC-MS/MS)测定方法,测定阿霉素纳米胶束和阿霉素注射液在大鼠体内的组织分布。方法采用甲醇∶生理盐水(4∶1)作为组织匀浆介质;色谱柱:SunFire C_(18)(250 mm×4.6 mm,5μm);流动相:5 mmol·L~(-1)醋酸铵-乙腈-甲酸(70∶30∶0.5);流速:0.5 mL·min~(-1);对乙酰氨基酚作为内标。在此色谱条件下,以多反应监测(MRM)扫描方式进行检测,将大鼠随机分成2组,分别测定大鼠尾静脉注射阿霉素纳米胶束和阿霉素注射液后各组织中阿霉素的药物质量浓度,给药剂量均为1 mg·kg~(-1)。结果大鼠各组织中阿霉素的质量浓度在0.5～200 ng·mL~(-1)范围内线性关系良好,r=0.999 1,样品的日内精密度RSD值小于6.7%,日间精密度RSD值小于13.2%。结论该方法快速、准确,适用于阿霉素在大鼠体内组织分布的研究,阿霉素纳米胶束与阿霉素注射液在心、脾和肺中的分布差异无统计学意义,在肝和肾中的分布差异有统计学意义,2种制剂均不能透过血脑屏障。     

超高效液相色谱-串联质谱法测定大鼠血浆及组织中贝达喹啉浓度及其药动学考察

目的:建立超高效液相色谱-串联质谱(UPLC-MS/MS)法测定SD大鼠血液及组织中贝达喹啉的浓度,研究其在大鼠体内的药动学及组织分布.方法:采用UPLC-MS/MS联用技术,测定大鼠血液及组织中贝达喹啉的药物浓度,并利用DAS软件及非房室模型统计矩方法计算药动学参数.结果:贝达喹啉线性范围为0.1～500 ng·mL...     

LC-MS/MS法研究氢溴酸东莨菪碱口腔速崩微囊片在比格犬体内的药代动力学

研究氢溴酸东莨菪碱口腔速崩微囊片在比格犬体内的药代动力学和生物等效性。液相色谱分离采用C18柱(100 mm×3.0 mm,3.5μm),甲醇-2 mmol.L-1甲酸铵溶液(25∶75)为流动相;质谱检测采用ESI离子源,正离子MRM方式检测。将6只比格犬随机分成2组,分别单剂量灌服氢溴酸东莨菪碱口腔速崩微囊片(受试制剂)和氢溴酸东莨菪碱普通片(参比制剂)0.6 mg,定时采集血样,测定比格犬体内的血药浓度并计算药代动力学参数。受试制剂和参比制剂的Cmax分别为(8.16±0.67)、(3.54±0.64)ng.mL-1;t1/2分别为(2.83±0.45)、(3.85±0.82)h;tmax分别为(1.25±0.27)、(0.42±0.09)h;AUC0-12 h分别为(25.06±3.75)、(9.59±1.02)h.ng.mL-1;AUC0-∞分别为(26.30±3.92)、(10.80±1.45)h.ng.mL-1;MRT0-12 h分别为(3.38±0.34)、(3.86±0.26)h;MRT0-∞分别为(3.98±0.63)、(5.37±1.00)h。受试制剂和参比制剂在AUC和吸收速率方面具有...     

UPLC-MS/MS法测定大鼠血浆及脑组织中的劳拉西泮和劳拉西泮甘氨酸酯

目的 采用超高效液相色谱-串联质谱技术(UPLC-MS/MS)同时测定大鼠血浆及脑组织中的劳拉西泮(LZP)和其新型前药劳拉西泮甘氨酸酯(LZP-Gly),并将其用于研究透皮离子导入LZP-Gly后大鼠体内的血浆药动学和药物的脑组织分布.方法 采用BEH C18色谱柱(50.0 mm×2.1 mm,2.5 μm),流动...     

乙烷硒啉在大鼠和结肠癌荷瘤裸鼠体内的组织分布研究

目的:考察抗肿瘤新药乙烷硒啉在大鼠和结肠癌荷瘤裸鼠体内的组织分布规律。方法:采用荧光分光光度法和液相色谱-质谱联用法分别测定SD大鼠和人结肠癌LS174T细胞荷瘤裸鼠灌服乙烷硒啉后各组织中药物浓度经时变化。结果:大鼠灌服乙烷硒啉2 h后,药物广泛分布于各组织,以胃肠及其内容物、肝、肾中含量最高;24 h肝肾组织硒浓度分别为(6.14±0.87)和(6.93±0.94)μg.g-1,同时间点血液中硒浓度为(4.43±1.65)μg.mL-1;48 h后除肾脏外,其他各组织硒浓度数值均低于血硒浓度。结肠癌裸鼠给药后1 h即可见原形药迅速分布于各组织,至24 h肾脏药物浓度达峰值,而其他组织中药物浓度显著降低,至48 h肿瘤中仍可检测出较高药物浓度。结论:乙烷硒啉口服给药后在大鼠和结肠癌荷瘤裸鼠体内均可广泛分布,并对肿瘤组织具有一定选择性,其组织分布特点可能与药物的代谢排泄途径相关。     

RP—HPLC法研究间尼索地平2种晶型在大鼠体内的组织分布特征

目的:采用高效液相色谱法研究2种晶型间尼索地平在大鼠体内的组织分布特征。方法:生物样品在碱性条件下,加入内标尼莫地平,经乙酸乙酯提取后,进行 HPLC 测定。色谱柱为 Diamonsil~(TM)C_(18)柱(250 mm×4.6 mm,5μm);流动相为乙腈-水(62:38),流速1.0 mL·min~(-1);检测波长237 nm。结果:大鼠在口服 A 和 B 两种晶型间尼索地平(30 mg·kg~(-1))后,于10,30,150 min处死,两种晶型在30 min 时组织中浓度较高,150 min 时在除肺外的各脏器中浓度均呈下降趋势。组织匀浆中间尼索地平的线性关系、最低检测限、准确度和精密度均符合分析要求。结论:该法测定大鼠各组织中药物浓度准确、灵敏、可靠,为问尼索地平新药研究与开发提供实验依据。     

高效液相色谱-质谱联用法测定大鼠血浆中五味子醇乙的浓度及其应用

目的 建立快速、灵敏的高效液相色谱-质谱法测定大鼠血浆中五味子醇乙的浓度,并研究其在大鼠体内的药动学变化规律.方法 以地西泮为内标,血浆样品经乙醚萃取后进行高效液相色谱-质谱分析,采用Hypersil-C18色谱柱(150mm×4.6mm,5μm),甲醇-水[72:28)为流动相,检测离子为399.00(五味子醇乙[M+H-H2O]+),284.90(内标地西泮[M+H]+.结果 血浆标准曲线线性范围为5·0～250 ng/mL(r=0.995 7),最低定量下限为5.0 ng/mL.五味子醇乙的提取回收率不小于78.80%,高、中、低3种浓度的日内、日间精密度RSD均小于15%(n=6),结论该方法选择性强、灵敏度高,适用于大鼠血浆中五味子醇乙浓度测定和临床药动学研究.     

HPLC法测定氢溴酸右美沙芬分散片中氢溴酸右美沙芬的含量

目的:建立高效液相色谱法测定氢溴酸右美沙芬分散片中氢溴酸右美沙芬的含量.方法:采用高效液相色谱法,色谱柱为TIAMHE○R C 18(200 mm×4.6 mm,5μm);流动相为甲烷磺酸溶液-乙腈(70:30);检测波长:280 nm,柱温:30℃;流速:1.0 ml/min.结果:氢溴酸右美沙芬在49.5~346.5μg/ml的浓度范围内,其浓度与峰面积呈良好线性关系(r=0.9999).平均回收率为100.05%,RSD为0.87%.结论:高效液相色谱法简便,快捷,精密度高,可用于氢溴酸右美沙芬分散片中氢溴酸右美沙芬含量测定.     

大鼠血浆中阿霉素HPLC-MS/MS测定法及其在药动学中的应用

目的建立大鼠血浆中阿霉素浓度的液相色谱-串联质谱(LC-MS/MS)测定方法。方法色谱柱:Venusil ASB C18(150 mm×4.6 mm,5μm),流动相:乙腈-5 mmol.L-1乙酸铵-甲酸(体积比为35.0∶65.0∶0.5),采用沉淀蛋白法,以多反应监测(multiple reaction monitoring,MRM)扫描方式检测,测定大鼠尾静脉注射阿霉素纳米胶束溶液后血浆中药物浓度。结果血浆中阿霉素质量浓度在1～1 000μg.L-1内线性关系良好,r=0.996 0;日内和日间精密度RSD≤13.2%;阿霉素的平均提取回收率为89.7%～95.7%;阿霉素在大鼠血浆中主要药动学参数为t1/2(30.5±5.2)h,ρmax(954.3±80.6)μg.L-1,AUC0-∞/(290.2±40.4)μg.h.L-1。结论该方法适用于阿霉素在大鼠体内药动学的研究。     

Pharmacokinetic and tissue distribution studies of paeoniflorin and albiflorin in rats after oral administration of total glycosides of paeony by HPLC-MS/MS

A simple, sensitive and specific high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method combined with solid phase extraction has been developed and validated for the simultaneous quantification of paeoniflorin and albiflorin in rats plasma and tissue homogenate after administration of total glycosides of paeony (TGP). Chromatographic separation was achieved on a C₁₈column (150 mm×4.6 mm, 5 μm) with mobile phase consisted of acetonitrile–0.05% formic acid aqueous (20: 80, v/v) at a flow rate of 0.5 mL/min. The analytes were detected with triple-quadrupole mass spectrometer using turbo ion spray with negative ionization in the multiple reaction-monitoring (MRM) modes. The results of method validationconformed to the requirements for the determination of biological samples. This method was successfully applied to the pharmacokinetics and tissue distribution study of TGP in rats. The results showed that paeoniflorin and albiflorin were absorbed and reached peak value quickly, and had the similar pharmacokinetic characteristics. Both of them underwent a rapid and wide distribution in the tissues throughout the whole body, among which stomach was the main distribution tissue. Moreover, they had the ability to cross the blood-brain barrier.     

UPLC-MS/MS法测定氘代氯化琥珀胆碱在大鼠体内分布

目的:建立超高效液相色谱串联质谱(UPLC-MS/MS)法测定氘代氯化琥珀胆碱(2H-Suc)在大鼠主要脏器分布。方法:大鼠皮下注射2H-Suc 4 h后,用UPLC-MS/MS法测定血清和相关脏器中2H-Suc含量;色谱柱:Luna NH2色谱柱(2 mm×100 mm,3μm),流动相为0.1%甲酸水溶液-乙腈等度洗脱,流速为0.2 mL/min;正离子扫描模式,多反应监测模式下,电喷雾离子化源进行检测;考察该方法的线性关系与检测限、准确度和精密度、回收率、基质效应和稳定性。结果:血清和肾组织中2H-Suc的检测限分别是0.04 ng/mL和0.05 ng/mL,线性范围分别是0.3~100 ng/mL和0.5~100 ng/mL,日内、日间精密度和准确度均小于15%,回收率在85.30%~96.76%之间,经过处理后的样品不存在明显的基质效应。大鼠皮下注射2H-Suc,在血清及心、肝、肾等主要脏器中均检测到2H-Suc,其中肾脏中含量最高。结论:UPLC-MS/MS法是测定大鼠体内2H-Suc含量的一个有效方法,2H-Suc在大鼠血清及心、肝、肾等中均有分布,肾脏中含量最高。     

达原饮中芍药苷和黄芩苷在大鼠体内药动学研究

目的:建立同时测定大鼠血浆中芍药苷和黄芩苷浓度的高效液相色谱-串联质谱(LC-MS/MS)方法,并研究大鼠体内达原饮的药代动力学。方法:采用蛋白质沉淀法处理血浆样品,色谱柱为Inersil ODS柱(4.6 mm×50 mm,5μm),流动相为水-乙腈梯度洗脱,柱温为35℃;流速为0.5 mL·min^-1。质谱采用电喷雾离子源(ESI),选择离子反应监测(SRM)模式。结果:该方法准确度、精密度、回收率和基质效应均符合生物基质样品测试要求,芍药苷和黄芩苷的线性范围均为5~2000 ng·mL^-1。芍药苷和黄芩苷的药代动力学参数AUC为(2.300374×10^4±7.42703×10^3)、(2.3423881×10^5±6.678954×10^4)ng·mL^-1·min;T1/2z为(126.65±34.65)、(168.18±46.44)min;Tmax为(30.00±0)、(27.50±6.12)min;Cmax为(170.44±53.76)、(1645.42±464.35)ng·mL^-1。结论:建立的大鼠血浆中芍药苷和黄芩苷浓度测定的LC-MS/MS方法简便易行,准确灵敏,适用于达原饮在大鼠体内的药代动力学研究。     

Pharmacokinetics and tissue distribution of 8,9-epoxy brevifolin in rats, a hepatoprotective constituent isolated from Phyllanthus simplex Retz by liquid chromatography coupled with mass spectrometry method

8,9-Epoxy brevifolin (EBF) is a novel compound isolated from Phyllanthus simplex Retz (P. simplex) and has been demonstrated to possess a hepatoprotective effect. The purposes of the present study were to examine the in vivo pharmacokinetics and tissue distribution of EBF in rats using a liquid chromatography coupled with mass spectrometry quantitative detection method (LC-MS/MS), with luteolin-7-O-glucoside being employed as an internal standard (IS). The method was validated within the concentration range 20-15 000 ng/ml, and the calibration curves were linear with correlation coefficients of >0.999. The limit of quantitation (LOQ) for EBF was 20 ng/ml. The intra-assay accuracy and precision ranged from 98.7% to 100.2% and 2.19% to 6.25%, respectively, while the inter-assay accuracy and precision ranged from 97.5% to 100.3% and 3.35% to 7.28%, respectively. The method was further applied to assess the pharmacokinetics and oral bioavailability of EBF after intravenous and oral administration in rats. The oral bioavailability of EBF was 12.46 +/- 2.31%. In the tissue distribution assay, its concentration was higher in the heart (13.2 +/- 0.24 microg/g) and liver (14.5 +/- 0.19 microg/g) than in other tissues.     

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.     

吲哚美辛5-氟尿嘧啶甲酯的代谢物5-氟尿嘧啶在大鼠体内组织分布和排泄

采用高效液相色谱法对吲哚美辛5-氟尿嘧啶甲酯(indomethacin 5-fluorouracil-1-ylmethyl ester，IFM)的代谢物5-氟尿嘧啶在大鼠体内的分布和排泄进行研究。生物样品经液-液萃取后，以甲醇-水-36%乙酸(3∶96.9∶0.1，v/v)或甲醇-水-36%乙酸(10∶89.9∶0.1，v/v)为流动相，使用Diamonsil^TM C₁₈色谱柱进行分离，检测波长为260 nm。大鼠灌胃IFM 100 mg·kg^-1后， 5-氟尿嘧啶在胃、 小肠、 肝组织中浓度较高，其他组织和血浆中较低。5-氟尿嘧啶从尿液(0～24 h)、粪便(0～48 h)中排泄量分别占给药总量的0.006 5%和0.063%。该方法准确、专属性强，适用于IFM代谢物5-氟尿嘧啶的临床前药代动力学研究。     

多柔比星白蛋白微球的生物性质及其在大鼠体内的分布

目的:研究多柔比星(DR)白蛋白微球生物性质及其在大鼠体内的分布。方法:采用胰酶降解法研究微球的生物降解性能并确定交联剂浓度及固化时间与其降解时间的关系;应用透析法研究DR白蛋白微球和DR水溶液的体外溶出/释药性;测定大鼠尾静脉分别注射DR白蛋白微球和DR水溶液2.5 mg·kg~(-1)后体内各组织中分布浓度。结果:降解时间随交联剂浓度和交联时间增加而延长,固化时间以90 min为宜;DR水溶液在6 h内溶出完全,微球混悬液在168 h释放量仅达80%;微球组的DR在肝和脾中分布浓度显著高于水溶液组(P<0.01),其它组织中则低于水溶液组。结论:DR白蛋白微球具有明显的缓释作用,对肝和脾表现出显著靶向性,对心脏具有很低的亲和性。     

Determination of TA-0201, a novel orally active non-peptide endothelin antagonist, in rat plasma and tissues by a liquid chromatography--electrospray ionization--tandem mass spectrometry system

N-[6-[2-[(5-bromo-2-pyrimidinyl)oxy]ethoxy]-5-(4-methylphenyl)-4-pyrimid inyl]-4-(2-hydroxy-1, 1-dimethylethyl) benzenesulfonamide sodium salt (TA-0201) is a novel orally active non-peptide antagonist for endothelin (ET) receptors. A sensitive and simultaneous determination method of TA-0201 and its major metabolites by liquid chromatography tandem mass spectrometry (LC--MS/MS) in a selected reaction monitoring mode using [2H6]TA-0201 as the internal standard was developed, and the plasma and tissue concentrations of TA-0201 and its major metabolites were determined in a pharmacokinetic study. The lower limit of determination of plasma TA-0201 concentrations by this method was 0.1 ng/0.5 ml, and the between- and within-run accuracy and precision were both less than 5.1%, in the calibration curve range of 0.1-50 ng/0.5 ml. This method was applied to determine the concentrations of TA-0201 and its metabolites in plasma and various target tissues, i.e. the heart, lung and kidney, after oral administration of TA-0201 (0.1 mg/kg(-1)) to male rats. TA-0201 and its major metabolite of a carboxylic acid form were detected in plasma and all the tissues 24 h after administration, their tissue concentrations being higher than those in plasma and still detectable at 72 h. Thus, this method could successfully be applied to study pharmacokinetic properties of TA-0201 in rats.     

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。     

Determination of gemifloxacin in different tissues of rat after oral dosing of gemifloxacin mesylate by LC–MS/MS and its application in drug tissue distribution study

A simple, sensitive and specific liquid chromatography–tandem mass spectrometry (LC–MS/MS) method was developed and validated to evaluate the accumulation of gemifloxacin in different tissues of Wister albino rat. The analytical method consists of the homogenization of tissues followed by simple liquid–liquid extraction and determination of gemifloxacin by an LC–MS/MS. The analyte was separated on a Peerless basic C₁₈ column (33 mm × 4.6 mm, 3 μm) with an isocratic mobile phase of methanol–water containing formic acid (1.0%, v/v) (9:1, v/v) at a flow rate of 0.6 ml/min. The MS/MS detection was carried out by monitoring the fragmentation of m/z 390.100 → 372.100 for gemifloxacin and m/z 332.100 → 314.200 for ciprofloxacin (internal standard; IS) on a triple quadrupole mass spectrometer. The validated method was accurate, precise and rugged with good linearity in all tissue homogenates. The accuracy and precision value obtained from six different sets of quality control samples of all tissues and serum analyzed in separate occasions within 91.833–102.283% and 0.897–5.291%, respectively. The method has been successfully applied to tissue distribution studies of gemifloxacin. The present study demonstrates that the highest tissue concentration of gemifloxacin was obtained in lung (11.891 ng/g), followed by liver (10.110 ng/g), kidney (10.095 ng/g), heart (4.251 ng/g), testis (3.750 ng/g), stomach (3.182 ng/g), adipose tissue (1.116 ng/g) and brain (0.982 ng/ml) in 3 h after multiple oral dosing of 200 mg gemifloxacin mesylate for 7 days. This method may also be used for gemifloxacin tissue distribution modeling study in rat tissues and antibiotic residue analyses in other animal tissues.