柱切换HPLC法同时测定人血浆中阿莫西林和克拉维酸浓度

柱切换 HPL C法同时测定人血浆中阿莫西林和克拉维酸浓度。血浆样品加头孢羟氨苄为内标 ,用乙腈沉淀蛋白及二氯甲烷反洗 ,水相进样后经在线净化 ,由流动相洗脱切入 C1 8分析柱进行分离测定。阿莫西林和克拉维酸线性范围分别为 0 .5～ 16和 0 .2 5～ 5 μg/ ml,净化回收率分别为 92 .8%～ 95 .2 %和 77.3%～80 .8% ,方法回收率分别为 94 .0 %～ 96 .1%和 88.9%～ 97.6 % ,日内 RSD分别为 0 .6 3%～ 1.6 1%和 1.4 5 %～ 2 .94 % ,日间 RSD2 .30 %～ 5 .31%和 5 .0 2 %～ 7.37%。本法准确可靠 ,已应用于阿莫西林克拉维酸钾分散片人体药动学研究     

全自动柱切换高效液相色谱系统测定人血浆中奥卡西平的浓度

目的采用全自动柱切换HPLC系统建立人血浆中奥卡西平的测定方法,用于临床治疗药物监测。方法全自动柱切换HPLC由萃取色谱系统及分析色谱系统2个部分构成,萃取柱为ASTON-RG C8柱(4.6 mm×50 mm,5μm,ANAX),分析柱为ASTON-RG C18柱(4.6 mm×250 mm,5μm,ANAX);2个系统间通过中心切割模式进行目标物转移,转移时间窗口2.6～4.2 min;聚焦流速为2.0 mL min-1,聚焦时间为0～0.6 min;检测波长305 nm;进样量100μL;采用外标法计算结果。结果标准曲线在0.47～26.07μg mL-1呈良好线性关系,相关系数为0.999 6,最低检测限为0.12μg mL-1,低(1.42μg mL-1)、中(11.85μg mL-1)、高(26.07μg mL-1)浓度的绝对回收率为95.6%～98.9%,日内和日间精密度均<5.0%,准确度99.6%～102.7%。结论本方法抗干扰能力强、自动化程度高、准确度和灵敏度高等优点,适合奥卡西平治疗药物监测的常规应用。     

柱切换高效液相色谱法测定人血浆中阿莫西林浓度

目的:建立柱切换高效液相色谱(HPLC)法测定人血浆中阿莫西林浓度.方法:血浆样品用乙腈沉淀蛋白及二氯甲烷反洗后,其水相进样于C18短柱,经0.02 mol·L-1磷酸二氢钾(pH3.0)在线净化,再由流动相0.02 mol·L-1磷酸二氢钾(pH4.6)-甲醇(95:5)洗脱切入C8分析柱进行分离测定.结果:线性范围0.2～20 mg·L-1,净化回收率81.7%～88.5%,方法回收率为93.2%～96.7%,日内RSD为1.25%～2.57%,日间RSD为2.74%～9.49%.结论:本法准确可靠,可用于阿莫西林体内血药浓度测定.     

HPLC同时测定人血浆中4种苯二氮类药物的浓度

目的 采用HPLC同时测定人血浆中地西泮、氯硝西泮、三唑仑、艾司唑仑的浓度.方法 血浆样品经氢氧化钠碱化后用乙酸乙酯萃取,氮气吹干,残留物用甲醇溶解后进样.采用Alhech-C18色谱柱,流动相为乙腈-甲醇-水(30:30:40),流速为1.0 ml·min-1,检测波长为225 nm.结果 三唑仑0.05-10.00 mg·L-1与峰面积的线性关系良好(r=0.9998),地西泮、艾司唑仑和氯硝西泮0.1～20.0 mg·L-1与峰面积的线性关系良好(r=0.9997).4种成分的方法回收率分别为96.8%、92.6%、97.7%、94.8%;13内和日间RSD均小于10%(n=5).结论 所建方法操作简便、快速、准确、灵敏度高、重复性好,适用于临床上人血浆中4种苯二氮(艹卓)类药物的同时定性、定量检测.     

血浆中去甲右美沙芬和愈创木酚甘油醚的HPLC测定

建立了灵敏度高、选择性好、可同时测定人血浆中右美沙芬活性代谢物去甲右美沙芬及愈创木酚甘油醚的HPLC-荧光检测法.采用C8柱(4.6mm×250mm),流动相为乙腈-0.05mol/L磷酸二氢钾-二乙胺溶液-冰醋酸(15:85:0.2:2,用磷酸调至pH3.0),检测波长λex=280nm,λem=313nm.以盐酸曲马多为内标,血浆样品经液-液萃取后进样测定.方法的日内、日间RSD小于4.63%,回收率97.3%～102.6%.去甲右美沙芬及愈创木酚甘油醚的检测灵敏度分别为5.412和20ng/ml.经方法学验证,满足药物动力学研究的要求.     

HPLC法同时测定人血浆中氢氯噻嗪与奥美沙坦浓度

目的建立同时测定人血浆中氢氯噻嗪、奥美沙坦浓度的HPLC法。方法采用Kromasil C18柱,以乙腈-0．02mol·L^-1 KH2PO4（32：68,V/V;磷酸调pH2．40）为流动相,坎地沙坦为内标,检测波长为272nm,血浆样品采用盐酸酸化后用EO基叔丁基醚提取浓集,按内标法定量。结果氢氯噻嗪血药浓度在0．005—0．64mg·L^-1内、奥美沙坦血药浓度在0．01～4．0mg·L^-1内线性关系良好（r=0．9991、r=0．9988）。氢氯噻嗪日内精密度（RSD）为3．11％～7．29％,日间RSD为3．72％～10．54％;奥美沙坦日内RSD为0．97％～5．18％,日间RSD为5．37％～9．17％。氢氯噻嗪提取回收率为78．38％～80．77％,相对回收率为98．01％．101．26％;奥美沙坦提取回收率为72．28％～74．35％,相对回收率为97．04％～100．17％。结论本法简便、灵敏、准确、重现性好,可用于人血浆中氢氯噻嗪、奥美沙坦浓度的测定。     

高效液相色谱法同时测定人血中对乙酰氨基酚和咖啡因含量

目的:建立同时测定人血浆中对乙酰氨基酚和咖啡因的反相高效液相色谱(RP-HPLC)方法.方法:以茶碱为内标,色谱分离采用Zorbax-Sb-C18分析柱(4.6 mm× 250 mm,5μm),柱温40℃,以乙腈-甲醇-缓冲液(7:7:86,缓冲液内含甲酸0.2%、甲酸铵18 mmol·L-1)为流动相;流速:1.0 mL·min-1,进样量:20μL;紫外检测波长:273 nm.结果:血浆中对乙酰氨基酚和咖啡因测定方法的线性范围分别为0.125～8 mg·L-1,0.0125～0.8 mg·L-1;方法回收率分别为99.76%～105.5%,98.35%～114.45%;日内精密度RSD分别小于3.0%,5.0%;日间精密度RSD分别小于等于12.0%,10.0%.结论:本方法操作简便,选择性好,准确、灵敏,可用于对乙酰氨基酚和咖啡因的人体药动学研究.     

阿昔洛韦血浆浓度的自动柱切换高效液相色谱法测定

目的建立适应大批量样品快速自动分析的阿昔洛韦血药浓度HPLC测定方法。方法应用自制自动磁力定时切换装置。分析柱采用WatersNova-Pak C     

高效液相色谱法测定炎痛净乳膏中双氯芬酸钠和苯佐卡因的含量

目的：测定炎痛净乳膏中双氯芬酸钠和苯佐卡因的含量。方法：高效液想象以谱法，甲醇为提取溶剂，地西泮为内标，No-va-PakC18色谱柱，甲醇-水-冰醋酸（80：20：0.5)为流动相，检测波长为283nm。结果：双氯芬酸钠和苯佐卡因在5μg/ml-40μg/ml范围内，其浓度与峰面积均呈良好的线性关系（r=0.9999)，平均回收率分别为101.0%,RSD=1.21%,99.8%,RSD=0.62%。结论：本法专属性强，操作简便，结果准确。     

Simultaneous determination of aceclofenac and its three metabolites in plasma using liquid chromatography-tandem mass spectrometry

A new LC/MS/MS-based method allows simultaneous determination of aceclofenac and its three metabolites (4'-OH-aceclofenac, diclofenac, and 4'-OH-diclofenac) in plasma. After acetonitrile-induced precipitation of proteins from the plasma samples, aceclofenac, 4'-OH-aceclofenac, diclofenac, 4'-OH-diclofenac, and flufenamic acid (an internal standard) were chromatographed on a reverse-phase C(18) analytical column. The isocratic mobile phase of acetonitrile/0.1% formic acid((aq)) [80:20 (v/v)] was eluted at 0.2 mL/min. Quantification was performed on a triple-quadrupole mass spectrometer employing electrospray ionization, and the ion transitions were monitored in multiple reaction-monitoring mode. The monitored transitions for aceclofenac, diclofenac, 4'-OH-diclofenac, 4'-OH-aceclofenac and flufenamic acid were m/z 352.9-->74.9, 296.1-->251.7, 311.8-->267.7, 368.9-->74.9, and 279.9-->235.9, respectively. The coefficient of variation of the assay precision was less than 6.5%, and the accuracy ranged from 93% to 103%. The limits of detection were 2 ng/mL for aceclofenac and 0.2 ng/mL for both diclofenac and 4'-OH-diclofenac. This method was used successfully to measure the concentrations of aceclofenac and its three metabolites in plasma from healthy subjects after administration of a single 100-mg oral dose of aceclofenac. This analytic method is a very simple, sensitive, and accurate way to determine the pharmacokinetics of aceclofenac and its metabolites.     

High performance liquid chromatographic determination of diclofenac sodium in plasma using column-switching technique for sample clean-up

For routine analysis of diclofenac sodium in plasma, a new high performance liquid chromatographic method, which is combined with column-switching technique is developed. The precolumn packed with Corasil RP C-18 was connected to analytical column by switching system in order to enrich the sample drugs in plasma without extraction. This method showed excellent sensitivity, precision and reproducibility. The limit of detection, using a 100 microL injection of plasma, was 0.1 micrograms/mL and the mean coefficient of variation for intra- and inter-assay was better than 4.6%. Total analysis time was 20 min between injections. The present method offers distinct practical advantages over conventional liquid-liquid extraction methods of sample preparation with respect to time, effort, recovery, and sample volume required. The method has been applied to the samples from rats receiving oral administration of diclofenac sodium.     

HPLC法同时测定人血浆中紫杉醇、多西他赛及环磷酰胺的浓度

目的建立一种快速同时测定人血浆中紫杉醇、多西他赛和环磷酰胺浓度的检测方法。方法采用双波长HPLC法,色谱柱为Shim-pack VP-ODS C₁₈(250 mm×4.6 mm,5μm);流动相为乙腈,水梯度洗脱;检测波长为紫杉醇、多西他赛230 nm,环磷酰胺197 nm;流速1 mL·min^-1。结果环磷酰胺、紫杉醇和多西他赛的保留时间分别为6.53、32.7和37.2 min,分别在血药浓度0.5~50、0.5~50、0.5~25 mg·L^-1内线性关系良好,方法回收率均≥95.0%;日内、日间精密度良好(RSD均<15%)。结论本方法灵敏、稳定、结果准确可靠,可用于紫杉醇、多西他赛和环磷酰胺的血药浓度测定。     

Simultaneous determination of amoxicillin and clavulanic acid in human plasma by isocratic reversed-phase HPLC using UV detection

A simple, rapid and sensitive isocratic reversed phase HPLC method with UV detection using internal standard has been developed and validated for simultaneous determination of amoxicillin and clavulanic acid in human plasma. The assay enables the measurement of amoxicillin and clavulanic acid for therapeutic drug monitoring with a minimum quantification limit of 15 and 30 ng ml(-1), respectively. The method involves simple, one-step extraction procedure and analytical recovery was complete. The separation was carried out in reversed-phase conditions using a Chromolith Performance (RP-18e, 100 mm x 4.6mm) column with an isocratic mobile phase consisting of 0.02 M disodium hydrogen phosphate buffer-methanol (96:4, v/v) adjusted to pH 3.0. The wavelength was set at 228 nm. The coefficients of variation for inter-day and intra-day assay were found to be less than 9.0%.     

Simultaneous determination of diclofenac and oxybuprocaine in human aqueous humor with HPLC and electrochemical detection

A sensitive and selective bioanalytical method for simultaneous determination of diclofenac and oxybuprocaine in human aqueous humor using reversed-phase HPLC and electrochemical detection is described. Chromatographic separation was achieved by using a Regis SPS 100 RP-8 column (5 μm; 150×4.6 mm I.D.). This support is coated with a hydrophilic polyoxyethylenepolymer. It allows protein-containing samples to be injected directly onto the column. The electrochemical detector permit a detection limit of 500 pg diclofenac per ml (daily relative standard deviation 6.3%) and 50 ng oxybuprocaine per ml (daily R.S.D. 2.6%), respectively. Results of administered and measured drug-concentrations in time dependent decrease are presented.     

HPLC法同时测定人血浆中甘草酸和苦参碱的质量浓度

目的建立以HPLC法同时测定人血浆中甘草酸和苦参碱质量浓度的方法。方法 Ultimate AQ-C18柱(250mm×4.6mm,5μm),柱温为室温(25℃),流动相为乙腈-三乙胺调pH至7.5的0.05mol·L-1磷酸二氢钾(22.5∶77.5),流速为1.0mL·min-1,检测波长为220nm(检测苦参碱)和250nm(检测甘草酸),进样量50μL。结果甘草酸单铵盐质量浓度在2.0²00.0μg·mL-1范围内线性良好,苦参碱质量浓度在2.0200.0μg·mL-1范围内线性良好,苦参碱质量浓度在2.0²00.0μg·mL-1范围内线性良好。平均回收率均大于95%,日内、日间RSD均小于4%。结论该方法简便、快速、灵敏、可靠,可用于人血浆中甘草酸和苦参碱质量浓度的监测。     

高效液相色谱法同时测定血浆中的氯胺酮、咪唑西泮的浓度

目的:建立同时测定血浆中氯胺酮、咪唑西泮浓度的高效液相色谱法.方法:采用Diamosil C18色谱柱(4.6 mm×150 mm,5μm),以甲醇-磷酸盐缓冲液(65:35)为流动相,非那西丁为内标,流速为1.0 mL·min-1,在220 nm波长下定量测定氯胺酮、咪唑西泮血浆浓度.结果:氯胺酮在0.25～10.0 mg·L-1浓度范围内线性良好,r=0.9994,平均回收率为99.33%,日内、日间精密度RSD为0.66%～3.66%.咪唑西泮在25～1000μg·L-1浓度范围内线性良好,r=0.9995,平均回收率为101.46%,日内、日间精密度RSD为1.48%～3.36%.结论:本文HPLC法操作简单,快速,重现性好,准确可靠,适用于临床血药浓度监测及药动学研究.     

反相高效液相色谱法同时测定人血浆中奎尼丁、利多卡因及普罗帕酮的浓度

目的 :建立反相高效液相色谱法同时测定人血浆中奎尼丁、利多卡因及普罗帕酮的浓度。方法 :应用UltrasphereC18柱(4.6mm× 2 5 0mm) ,流动相为甲醇 水 冰醋酸 三乙胺 (6 2 .3∶37.7∶0 .0 34∶0 .0 6 8) ,pH 6 .0 2 ,紫外 2 2 0nm检测。结果 :奎尼丁、利多卡因、普罗帕酮平均回收率分别为 97.3% ,98.9% ,99.0 % ,日内日间精密度分别为 (2 .11～ 6 .0 1) % ,(1.2 5～ 4.35 ) % ,(1.5 0～ 4.2 7) %。结论 :方法简单、灵敏、准确 ,可用于临床治疗药物监测。     

Simultaneous isocratic HPLC determination of vigabatrin and gabapentin in human plasma by dansyl derivatization

A rapid and low-cost assay for simultaneous vigabatrin (VGA) and gabapentin (GBP) determination is described that can be performed with simple HPLC instrumentation. The method involves derivatization of the primary amine group of VGA and GBP with dansyl chloride followed by isocratic separation (column: microBondapak C-18, 10 microm, 300 x 3.9 mm; mobile phase: 50 mmol/L NaH(2)PO(4) in 40% acetonitrile) at 50 degrees C and fluorometric detection (excitation and emission wavelength: 318 and 510 nm, respectively) of the fluorescent product, which is stable for at least 7 days. Correlation coefficients of the calibration curves are >0.999 with a lower limit of detection of 0.3 microg/mL. Between- and within-run coefficients of variation are below 4.5%, and assay time is 15 minutes. This method may be used for therapeutic drug monitoring in the case of GBP and to control patient compliance in the case of VGA.     

HPLC同时测定血浆中头孢氨苄和甲氧苄啶的浓度

采用高效液相色谱紫外检测器同时检测血浆中头孢氨苄和甲氧苄啶的浓度。以0.5倍血浆体积,浓度为10%(v/v)的高氯酸为蛋白沉淀剂。色谱条件:Diamonsil C18(250mm×4.6mm,5μm),柱温:30℃,流动相:pH3.0的0.05mol/L的磷酸盐缓冲液∶乙腈∶甲醇(76∶16∶8),流速1.0ml/min,进样量20μl,检测波长240nm。头孢氨苄、甲氧苄啶及内源性物质分离良好。头孢氨苄血药浓度线性范围为0.5~50.0μg/ml,检测限为50ng/ml,高、中、低浓度的相对回收率在97.3%~100.5%之间(n=5),日内RSD≤6.5%(n=5),日间RSD≤8.3%(n=5)。甲氧苄啶血药浓度线性范围为0.25~25.0μg/ml,检测限为25ng/ml,高、中、低浓度的相对回收率在96.5%~99.3%之间(n=5),日内RSD≤6.9%(n=5),日间RSD≤8.6%(n=5)。本法样品处理简单、灵敏、准确,适合于同时测定两者的体内浓度。     

Direct determination of stability of protease inhibitors in plasma by HPLC with automated column-switching

Automated column-switching HPLC methods were developed and utilized for the direct analyses of three hydroxamic acid based metalloprotease inhibitors in rat plasma. These column-switching methods involved the use of a restricted-access media (RAM) precolumn and a column-switching valve, allowing the complete automation of sample preparation and HPLC. The plasma samples were directly injected onto a precolumn packed with SPS/ODS stationary phase and then backflushed onto an ODS analytical column using a 6-port column-switching device. The drug stability in rat plasma was determined using both the automated and traditional HPLC methods. The results obtained from the automated column-switching methods were in good agreement with those from traditional methods that involve sequential protein precipitation, liquid extraction, solvent evaporation, and sample reconstitution. In addition to the elimination of labor-intensive and time-consuming sample preparation procedures, the column-switching methods allowed on-line analyte enrichment and accurate determination of drug stability in plasma with detection limits in the range of 10-20 ng/ml(-1). This work represents, for the first time, a drug stability study in plasma by automated column-switching HPLC technique with the use of a RAM column. Our column-switching methods can be readily adapted to any existing HPLC system with minimal hardware modification.