高效液相色谱法测定家兔血浆中右旋酮洛芬浓度

目的 :采用高效液相色谱法测定家兔血浆中右旋酮洛芬浓度。方法 :用乙醚提取血浆样品中右旋酮洛芬 ,在C18 柱上 ,以甲醇 -50mmol/L磷酸二氢钠 (30∶70)为流动相进行分离 ,于260nm波长处检测。结果 :线性范围为0 05～25μg/ml,回收率为99 92 %～100 54 % ;日内标准差为0 56 %～2 94 % ,日间标准差为0 6 %～2 89 %。结论 :本法简便、准确 ,重现性好 ,用于测定6只家兔口服右旋酮洛芬 -β-环糊精缓释微球血药浓度取得良好效果     

右旋兰索拉唑双相释放胶囊在Beagle犬体内的药动学研究

建立了LC-MS/MS法测定犬血浆中的右旋兰索拉唑,并研究了右旋兰索拉唑控释胶囊(商品名:Dexilant)在Beagle犬体内的药动学.采用C18柱,以甲醇-水(含10 mmol/L甲酸铵,70:30)为流动相,奥美拉唑为内标.右旋兰索拉唑在5～2 000 ng/ml浓度范围内线性关系良好,提取回收率105.7％～123.1％,批内、批间RSD均小于10％.Beagle犬口服给药后,呈明显血药双峰现象,药动学参数t1/2为(0.6±0.1)h,AUC0-∞为(2 019±176) ng·ml-1h,cmax为(810.9±194.0) ng/ml.     

HPLC法同时检测茶叶中八种儿茶素

建立一种茶叶中多组分儿茶素的HPLC测定方法。将茶叶用80℃热水超声提取20min,提取液离心分离后,采用Kromasil C18(250mm×4.6mm,5μm)色谱柱分离,以水-乙腈-磷酸(49.95:50:0.05,V:V)为流动相A,水-乙腈-磷酸(95.45:4.5:0.05,V:V)为流动相B,进行梯度洗脱,流速为0.5mL/min,柱温25℃,紫外检测波长231nm。在给定的浓度范围内,各儿茶素组分的峰面积与其相应浓度呈现良好的线性相关,R2>0.9950。添加已知儿茶素含量的回收试验结果显示,各组分儿茶素的平均回收率为94.8%～110.8%,相对标准偏差≤2%。     

犬血浆中右旋布洛芬和左西替利嗪的正负离子切换LC-MS/MS法测定

建立了液相色谱-串联质谱法同时测定Beagle犬血浆中的右旋布洛芬和左西替利嗪.使用C18色谱柱,以乙腈-甲醇-5mmol/L乙酸铵溶液(25∶56.25∶18.75)为流动相,采用电喷雾电离源、多反应监测模式,正负离子切换扫描,右旋布洛芬和左西替利嗪在0.2～100μg/ml和0.008～4μg/ml范围内线性关系良好,日内和日间RSD小于8.7％,提取回收率大于80.9％.     

HPLC法测定胃癌患者卡培他滨血药浓度及其药动学研究

目的:建立卡培他滨的血药浓度测定方法并对其在胃癌患者体内的药动学进行初步探讨。方法:入组4例胃癌患者,其血浆以替硝唑为内标,以叔丁基甲醚进行提取后进样测定,色谱柱为EclipseC18,流动相为乙腈-水(26∶74),检测波长为310nm,流速为1.0ml/min。血药浓度数据用DAS药动学计算机程序处理。结果:卡培他滨与内标替硝唑分离良好,保留时间分别为8.15、3.15min,卡培他滨血药浓度在0.05～8.00μg/ml范围内线性关系良好;日内、日间RSD均<7%,提取回收率在77.04%～87.53%范围内;卡培他滨的药动学参数cmax、tmax、t1/2β、AUC0-6h分别为(5.42±2.90)μg/ml、(1.0±0.71)h、(1.01±0.50)h和(9.54±5.32)mg·h/L。结论:胃癌患者口服卡培他滨后吸收迅速,消除半衰期短,个体之间生物利用度差异显著。     

翻白草中儿茶素的提取工艺研究

目的 研究翻白草中儿茶素的最佳提取工艺.方法 以儿茶素的提取含量为评价指标,采用超声波提取法,通过正交试验研究不同浓度乙醇、提取次数、提取时间、料液比4个因素对儿茶素提取的影响,并筛选出最佳提取条件;用高效液相色谱法测定含量,以甲醇-水-乙酸（25∶74.5∶0.5）作为流动相,流速为1.0 mL/min,检测波长为280 nm.结果 最佳提取条件为浓度30%的乙醇,料液比1∶15,提取3次,每次提取20 min;儿茶素的含量茎叶中为0.216 6 mg/g,根中为0.156 0 mg/g,平均回收率为98.52%,RSD为2.98%.结论 该方法简便、迅速,测定结果可为翻白草在临床用药中的安全性和有效性提供科学依据.     

熊胆降热丸中儿茶素在大鼠体内的药代动力学研究

目的研究熊胆降热丸中儿茶素在大鼠体内的药代动力学特性。方法以乙腈-水-三乙胺(6∶94∶0.3,磷酸调节pH=4.0±0.1)为流动相,血浆样品经甲醇-饱和硫酸锌(1∶1)除蛋白,采用高效液相色谱(HPLC)法测定儿茶素的浓度。利用DAS2.0软件计算儿茶素的主要药代动力学参数。结果儿茶素在0.12～9.6 mg/L范围内线性关系良好(r=0.999 2),平均绝对回收率为90.2%～97.1%,日内和日间RSD均低于15%。儿茶素大鼠体内的主要药代动力学参数Cmax为(1.670±0.754)mg/L,T1/2β为(7.779±4.568)h,AUC0-t为(6.192±2.828)mg/(L.h),AUC0-∞为(7.444±4.176)mg/(L.h)。结论该方法快速、准确、灵敏,可用于儿茶素的药代动力学研究,为儿茶药材和含儿茶中药复方制剂的体内研究和临床合理用药提供参考。     

高效液相色谱法测定血浆中洛美沙星浓度及其在人体内药代动力学

建立了人血浆洛美沙星的反相离子对高效液相色谱测定方法。该法简便易行,精密度好,方法回收率95～102%,日内、日间RSD为2.1～7.8%,血药浓度在0.125～5.012 μg/ml范围内呈线性关系,相关系数0.9998,当S/N=2时,最小检测浓度为25 ng/ml。健康志愿者口服400 mg洛美沙星,药代动力学过程符合一室模型,消除相半衰期为5.5 h。     

不同剂量钴-60辐照对制痂酊中儿茶素和表儿茶素含量的影响

目的:建立HPLC法测定制痂酊中儿茶素和表儿茶素含量的方法,同时考察不同剂量⁶⁰Co辐照对其含量影响。方法:色谱柱:Kromasil C₁₈（250 mm×4.6 mm,5μm）,流动相:2.2%冰醋酸-乙腈（86:14）,流速:0.8 ml·min^-1,检测波长:280nm,柱温:35℃。结果:儿茶素和表儿茶素分别在1.025～12.813μg,0.196～2.450μg范围内线性良好,加样回收率分别为98.0%,99.8%（n=5）;⁶⁰Co辐照剂量为3 kGy时,儿茶素和表儿茶素含量分别减少2.77%,3.43%,随着辐照剂量的增加,两者含量有逐渐下降的趋势。结论:对单批次样本测定表明:⁶⁰Co辐照对制痂酊中儿茶素和表儿茶素的含量有一定影响,但在低剂量（≤3 kGy）时影响不显著。     

高效液相色谱法测定乙吗噻嗪在人的血浆浓度及药代动力学

建立了反相高效液相色谱法测定人血浆中乙吗噻嗪浓度。色谱柱采用SpherisotbC18柱(25cm×4.6mm,5μm),流动相为甲醇—水—三乙胺(70:30:0.4,pH6.5),检测波长268nm。用乙腈沉淀蛋白后,吹干浓缩进样。血药浓度在20～4000ng/ml范围内呈线性关系,相关系数0.9994,血浆最低检测浓度3ng/ml。方法回收率90～103%,日内、日间RSD2.4～10.2%。应用该法研究了8名志愿者口服乙吗噻嗪片后的药代动力学,用一室模型拟合,消除相半衰期为1.75±0.45h。本法简便、回收率和灵敏度高、重复性好,适于临床药代动力学和药效学的研究。     

A rapid reversed phase high performance liquid chromatographic method for determination of etoposide (VP-16) in human plasma

A rapid, simple and sensitive isocratic High Performance Liquid Chromatography (HPLC) method was developed to measure the concentration of etoposide in plasma samples with UV detection at 220 nm. The method uses a Bondapac C18 column at 60 degrees C. The mobile phase consists of Methanol: water (45:55 v/v) at a flow rate of 2.8 ml/min. Phenacetin was used as an internal standard. The plasma samples were extracted using ether with the organic layer evaporated under nitrogen. The residue was dissolved in 200 microl methanol with 20 microl injected into the HPLC column. The extraction method showed a recovery of 91.5+/-3% for etoposide. In this system, the retention time of phenacetin and etoposide were 3.3 and 4.4 min, respectively. The limit of detection of etoposide in plasma is 20 ng/ml and the limit of quantitation is 40 ng/ml. This analytical method has very good reproducibility (8.1% between-day variability at a concentration of 50 ng/ml). It is a fast, sensitive and economic method applicable for clinical and pharmacokinetic studies.     

HPLC assay for albendazole and metabolites in human plasma for clinical pharmacokinetic studies

A sensitive and selective HPLC chromatography method using UV detection (295 nm) was developed for the determination of albendazole, albendazole sulfoxide (ABZSO), and albendazole sulfone (ABZSO2) in human plasma. Albendazole, ABZSO, ABZSO2, and the internal standard, oxibendazole, were extracted from human plasma by loading onto a conditioned C(18) SPE cartridge, rinsing with 15% methanol, and eluting with 90% methanol. Samples were evaporated under a stream of nitrogen, reconstituted with mobile phase, 1.25% triethylamine in water-methanol-acetonitrile (72:15:13, v/v/v) (pH* 3.1), and injected onto a Waters muBondapak Phenyl 3.9 x 300 mm HPLC column. Mobile phase flow rate was 1.0 ml/min. The retention times of albendazole, ABZSO, ABZSO2, and the internal standard were approximately 24.4, 7.9, 13.4, and 11.3 min, respectively. Total run time was 30 min. The assay was linear for concentration ranges in human plasma of 20-600 ng/ml for albendazole, 20-1000 ng/ml for ABZSO, and 20-300 ng/ml for ABZSO2. The analysis of quality control samples demonstrated excellent precision. Coefficients of variation for albendazole (20, 400, 600 ng/ml) were 6.7, 8.1 and 7.0%; ABZSO (20, 400, 800 ng/ml) were 6.0, 8.5 and 5.9%; ABZSO2 (20, 150, 300 ng/ml) were 3.1, 3.9 and 2.3%, respectively. The method appears to be robust and has been applied to a pharmacokinetic study of albendazole in healthy volunteers.     

LC-ESI-MS method for the determination of bisoprolol in human plasma

A sensitive liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) method has been developed and validated for the determination of bisoprolol in human plasma, using metoprolol as internal standard (I.S.). After alkalization with sodium hydroxide, the samples were extracted with ethyl acetate and separated by HPLC on a ZORBAX SB-C18 column with a mobile phase of 10 mM ammonium acetate buffer containing 0.1% formic acid-methanol (32:68, v/v) at a flow rate of 1 ml/min. The chromatographic separation was achieved in less than 5 min. The linearity was established over the concentration range of 0.05-120 ng/ml. The intra- and inter-run standard deviation was less than 3.8 and 7.5%, respectively. The method had been successfully applied to study the relative bioavailability of bisoprolol fumarate tablets in healthy Chinese volunteers. The pharmacokinetic parameters of the reference and test tablets have been compared.     

HPLC method for the determination of fluvoxamine in human plasma and urine for application to pharmacokinetic studies

A simple, specific and sensitive high-performance liquid chromatographic (HPLC) method has been developed for the assay of fluvoxamine in human plasma and urine. The method was based on reaction of fluvoxamine with 1,2-naphthoquinone-4-sulphonic acid sodium salt (NQS) forming orange colored product. The fluvoxamine-NQ derivative was separated by isocratic reversed-phase HPLC and detected at 450 nm. The chromatographic conditions were as follows: Phenomenex C(18) (250 mm x 4.6 mm i.d., 5 microm) column, mobile phase consisting of acetonitrile/water (80:20 v/v) at a flow rate of 1 ml/min. Tryptamine was selected as an internal standard. The assay was linear over the concentration range of 5-145 and 2-100 ng/ml for plasma and urine, respectively. The limits of detection (LOD) were 1.4 and 1 ng/ml for plasma and urine estimation at a signal-to-noise (S/N) ratio of 3. The limits of quantification (LOQ) were 5 and 2 ng/ml for plasma and urine, respectively. The extraction recoveries were found to be 96.66+/-0.69 and 96.73+/-2.17% for plasma and urine, respectively. The intra-day and inter-day standard deviations (S.D.) were less than 1. The method indicated good performance in terms of specificity, linearity, detection and quantification limits, precision and accuracy. This assay was demonstrated to be applicable for clinical pharmacokinetic studies.     

High-Performance liquid chromatographic analysis of lactone and hydroxy acid of new antitumor drug, DX-8951 (exatecan), in mouse plasma

A sensitive high-performance liquid chromatographic (HPLC) method has been developed and validated for the determination of lactone and total drug (lactone plus hydroxy-acid) of DX-8951 in mouse plasma. Solid-phase extraction by C18 cartridge separated lactone from total drug of DX-8951. Analysis was performed using a reverse-phase ODS column with a mobile phase consisting of acetonitrile/0.05 M potassium dihydrogen phosphate (pH 3) (18: 82, v/v) at a flow rate of 1 ml/min. The limits of quantitation of lactone and total drug were 3 ng/ml in plasma and a linear range of determination were observed over the concentration of 3 to 500 ng/ml. This method was applied to pharmacokinetic study in male mice treated with a single intravenous administration of either lactone or hydroxy-acid of DX-8951. The plasma concentrations of lactone from 2 to 6 h after dosing were similar regardless of the form of DX-8951 administered.     

柱前衍生化反相HPLC法测定血浆中普罗帕酮的对映异构体

建立了选择性测定血浆中普罗帕酮对映异构体的柱前衍生化反相HPLC法。用S(+)-1-(1-萘基)乙基异氰酸酯为衍生化试剂,与血浆中提取出的普罗帕酮反应生成非对映立体异构体,以HPLC-UV检测法(220nm)定量。采用此法成功地测试了10名健康受试者单剂量口服300mg盐酸普罗帕酮片后对映异构体的药代动力学曲线。此法灵敏度高(7.5ng·ml^-1),操作简便,重现性好。     

Liquid chromatographic assay of nifedipine in human plasma and its application to pharmacokinetic studies

A highly sensitive, selective and reproducible reversed-phase high-performance liquid chromatographic method has been developed for the determination of nifedipine in human plasma with minimum sample preparation. The method is sensitive to 3 ng/ml in plasma, with acceptable within- and between-day reproducibilities and linearity (r2 > 0.99) over a concentration range from 10-200 ng/ml. Acidified plasma samples were extracted using diethyether containing diazepam as internal standard and chromatographic separation was accomplished on C18 column using a mobile phase consisting of acetonitrile, methanol and water (35:17:48, v/v). The within-day precision ranged from 2.22 to 4.64% and accuracy ranged from 102.4-106.4%. The day-to-day precision ranged from 2.34-7.07% and accuracy from 95.1-100.1%. The relative recoveries of nifedipine from plasma ranged from 91.0-107.3% whereas extraction recoveries were 88.6-93.3%. Following eight 6-week freeze-thaw cycles, nifedipine in plasma samples proved to be stable with accuracy ranging from 0.64 to 3.0% and precision ranging from 3.6 to 4.15%. Nifedipine was also found to be photostable for at least 120 min in plasma, 30 min in blood and for 60 min in aqueous solutions after exposure to light. The method is sensitive and reliable for pharmacokinetic studies and therapeutic drug monitoring of nifedipine in humans after the oral administration of immediate-release capsules and sustained-release tablets to five healthy subjects.     

Development and validation of a LC-MS/MS method with electrospray ionization for determination of LASSBio-579 in rat plasma

A simple and sensitive LC-MS/MS analytical method was developed and validated for the determination of LASSBio-579 in plasma rat, using fluconazole as internal standard. Analyses were performed on a Shimadzu HPLC system using a Shimadzu C18 column and isocratic elution with acetonitrile-water (80:20, v/v), containing 0.4mM ammonium hydroxide and 0.2 mM acetic acid at a flow rate of 1.0 ml/min (split ratio 1:5). A Micromass triple quadrupole mass spectrometer, equipped with an electrospray ionization interface, operated in the positive mode. Plasma samples were deproteinized with acetonitrile (1:2) and 50 microl of the supernatant were injected into the system. The retention times of LASSBio-579 and IS were approximately 4.7 and 2.4 min, respectively. Calibration curves in spiked plasma were linear over the concentration range of 30-2000 ng/ml with determination coefficient >0.98. The lower limit of quantification was 30 ng/ml. The accuracy of method was within 15%. Intra- and inter-day relative standard deviations were less or equal to 13.5% and 6.4%, respectively. The applicability of the LC-MS/MS method for pharmacokinetic studies was tested using plasma samples obtained after intraperitoneal administration of LASSBio-579 to male Wistar rats. No interference from endogenous substances was observed, showing the specificity of the method developed. The reported method can provide the necessary sensitivity, linearity, precision, accuracy, and specificity to allow the determination of LASSBio-579 in pre-clinical pharmacokinetic studies.     

Validated high performance liquid chromatographic method for simultaneous determination of rosiglitazone, cilostazol, and 3,4-dehydro-cilostazol in rat plasma and its application to pharmacokinetics

A high performance liquid chromatographic (HPLC) method for simultaneous determination of rosiglitazone, CAS 122320-73-4, RSG), cilostazol (CAS 73963-72-1, CLZ) and its active metabolite 3, 4-dehydro-cilostazol (DCLZ), using pioglitazone (PIO) as internal standard (IS), in rat plasma is described. The plasma was extracted with methyl t-butyl ether, the dry extract was reconstituted in mobile phase and the aliquot was injected. The eluent drugs were detected by UV at dual wavelength of 226 nm (RSG and DCLZ) and 257 nm (CLZ). The mobile phase consisting of acetonitrile:potassium di-hydrogen phosphate buffer (35:65 v/v) was used at the flow rate of 1.2 ml/min on a reverse phase C18 column. The absolute recovery was above 90% of all analytes over the concentration range of 25-2500 ng/ml for RSG and CLZ and 20-2000 ng/ ml for DCLZ. The relative standard deviation (RSD) of the inter-day and intra-day precision ranged from 2.8 to 8.4% and 0.9 to 5.9%, respectively. The method is simple, rapid, accurate and sensitive and was applied to pharmacokinetic studies.     

Sensitive and selective liquid chromatography-mass spectrometry method for the quantification of rosiglitazone in human plasma

A sensitive and selective high-performance liquid chromatography-electrospray ionisation-tandem mass spectrometry (HPLC-ESI-MS-MS) method for determination of rosiglitazone in human plasma has been developed. After the addition of the internal standard, plasma samples were precipitated by acetonitrile. The compounds were separated on a proC18 column using a mixture of ammonium acetate buffer (0.02 M, pH 6.5) and acetonitrile (in the ratio of 47:53, v/v) as mobile phase. A Finnigan LCQdeca plus ion trap mass spectrometer connected to a Finnigan Surveyor HPLC was used to develop and validate the method. Linearity was established for the range of concentrations 1-1000 ng/ml with a coefficient of determination (r(2)) of 0.999. The intra-day accuracy for rosiglitazone ranged from 110.0 to 99.2% at low, medium and high levels. The inter-day accuracy was less than 15%. The lower limit of quantitation (LLOQ) was identified reproducible at 1.0 ng/ml with a precision of 5.7%. After validation, the method was used to study the pharmacokinetic profile of rosiglitazone in five healthy volunteers after administration of a single oral dose (4.0mg). The proposed method enabled the unambiguous evaluation and quantitation of rosiglitazone for pharmacokinetic, bioavailability or drug-drug interaction studies. A possible chromatography peak (m/z 121, its parent ion m/z 344) of N-demethyl rosiglitazone was observed at 3.49 min during determining rosiglitazone. This may be also a potential method for simultaneous determination of rosiglitazone and its metabolite N-demethyl rosiglitazone concentrations in plasma.