高效液相色谱法快速测定人血浆拉莫三嗪浓度

目的 建立快捷、灵敏、简便的高效液相色谱法(HPLC)测定拉莫三嗪(LTG)浓度,并应用于治疗药物浓度监测和药物相互作用研究.方法 以非那西丁为内标,检测波长为240 nm,使用Xterra MS C18色谱柱(4.6 mm×150 mm,5μm)为分析柱,以醋酸乙酯 二氯甲烷-叔丁基甲醚(3:1:1)为萃取剂,流动相为甲醇:水(含20 mM醋酸铵盐)(35:65,V/V),流速为1 ml/min.结果 LTG线性范围在0.1 ～ 50 μg/ml范围内线性良好(r ＞0.999),提取回收率为77.42％～83.05％,批间批内RSD均小于10％.应用HPLC检测使用LTG的87例癫痫患者血浆样本,对日剂量和血药浓度建立简单线性回归方程进行分析,血药浓度个体间差异大,剂量决定力不强(R2 =0.340).结论 HPLC简便、灵敏,可用于临床药物监测和研究,以辅助LTG个体化治疗.     

简单灵敏的HPLC法测定人血浆中头孢丙烯浓度

目的:建立血浆中头孢丙烯的HPLC检测方法.方法:采用高效液相色谱检测法,分析柱为Hypersil BDS C18 (200 mm×4.6 mm,5μm),流动相为20 mmol/L乙酸铵/10％乙酸(pH=3.7):乙腈=92:8(V/V),流速1.0 mL/min,柱温30℃,检测波长282 nm,进样量20μL.血浆样品用三氟醋酸直接沉淀后,取上清进样分析.结果:头孢丙烯在0.10～10.01 μg/mL范围内线性关系良好(r=1.0000),最低定量浓度为0.10μg/mL,日内日间RSD均＜15％(n=15).结论:研究建立的HPLC法快速、简便、高效,精密度和准确度高,适合血样中头孢丙烯浓度的检测.     

高效液相色谱法测定人血浆中拉莫三嗪的浓度

目的建立测定人血浆中拉莫三嗪浓度的高效液相色谱法。方法色谱柱为SunFire C18(4.6 mm×250mm,5μm),流动相为水-乙腈-三乙胺(64.5:35:0.5,v/v/v),磷酸调节pH至6.5,流速为1.0 mL·min-1,血浆标本经乙酸乙酯-二氯甲烷(75:25,v/v)提取后,在237 nm波长下进行检测。结果拉莫三嗪在0.312 5⁴0.0μg·mL-1与峰面积呈良好的线性关系(Y=0.047 2X+0.069 3,r2=0.999 2),相对回收率均在82%40.0μg·mL-1与峰面积呈良好的线性关系(Y=0.047 2X+0.069 3,r2=0.999 2),相对回收率均在82%¹05%,日内和日间RSD均<5%。结论该方法专属性强、准确度好、灵敏度高、操作简便,适合临床上对拉莫三嗪血药浓度的常规监测。     

简单灵敏的高效液相色谱法测定人血浆中去甲万古霉素的浓度

目的建立血浆中去甲万古霉素含量的HPLC测定方法。方法采用高效液相色谱检测法,分析柱为Tskgel ODS-80TS（4．6mm×250mm,5μm）,流动相为10mmol·L^-1甲酸铵（0．1％甲酸）-甲醇（0．1％甲酸）=（82：18,v／v）,流速：1．0mL·min^-1,柱温：30℃,检测波长：236nm,进样量：20μL.血浆样品用三氟醋酸直接沉淀后,进样分析。结果去甲万古霉素在1～1000μg·mL^-1与峰面积线性关系良好（r=0．999）,最低检测浓度为1μg·mL^-1。平均回收率为96％;日间、日内RSD均〈15％。结论研究建立的HPLC法快速、简便、高效,精密度和准确度高,适合血样中去甲万古霉素浓度的检测。     

A rapid, sensitive HPLC method for the determination of ganciclovir in human plasma and serum

A method for ganciclovir determination in human serum and plasma has been developed and validated. The method has a lower limit of quantification (LLOQ) adequate for sensitive pharmacokinetic studies ( < or = 0.05 microg/ml), has run times of < or = 15 min, and uses aliquot volumes adequate for pediatric studies (0.25 ml). In the method, proteinaceous material in serum or plasma is precipitated by trichloroacetic acid. An aliquot of the supernatant is analyzed by HPLC; automated column switching removes late-eluting materials that might interfere with the analyte peak in subsequent runs. Detection and quantification of ganciclovir is by fluorescence (lambda(ex) = 278 nm; lambda(em) = 380 nm). The method has a validated range of 0.0400-4.00 microg/ml and an LLOQ of 0.0400 microg/ml. All intra- and inter-assay % C.V. values were < 8%; all recoveries (accuracy) were within 7% of nominal values. No interference was observed by mycophenolic acid or its glucuronide metabolite, by AZT, salicylic acid, acetaminophen, ibuprofen, naproxen prednisone, acyclovir, or cyclosporine. Ganciclovir is very stable in the samples and the extract during storage and sample processing. Both serum and plasma methods have been validated for use and have been successfully used to analyze samples from clinical studies.     

A simple method to monitor plasma concentrations of oxcarbazepine, carbamazepine, their main metabolites and lamotrigine in epileptic patients.

A rapid, selective and sensitive isocratic reversed-phase HPLC method for the simultaneous determination of oxcarbazepine, its main metabolites (mono and dihydroxycarbazepine), lamotrigine, carbamazepine and carbamazepine-epoxide in plasma samples has been implemented. Analytes were extracted on solid-phase cartridges (SPE) and chromatographic separation was achieved on a Zorbax SB-CN column. The chromatographic peak area ratio based on UV absorbency at 214 nm was used for quantitative analysis. This HPLC method has been successfully used for routine evaluation to monitor plasma concentrations in epileptic patients referred to our institute and for pharmacokinetic studies regarding patients cotreated with drugs inducing or inhibiting OXC metabolism.     

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.     

高效液相色谱法同时测定人血浆中拉莫三嗪和奥卡西平活性代谢物血药浓度及其临床应用分析

目的：建立HPLC法同时测定人血浆中拉莫三嗪和奥卡西平活性代谢物血药浓度的方法，应用于临床常规检测，为患者个体化用药提供依据。方法：采用反相高效液相色谱法建立同时测定人血浆中拉莫三嗪（LTG）和奥卡西平活性代谢物（MHD）血药浓度的方法，方法学验证合格后用于治疗药物浓度监测（TDM），2013年8月-2016年3月，共完成1 611人次LTG和MHD血药浓度的监测，并对监测结果进行统计分析。结果：991人次（61.51%）在有效血药浓度范围内，13例患者进行了多次LTG血药浓度监测，达有效血药浓度的比例从61.54%上升至84.62%；23例患者进行了多次MHD血药浓度监测，达有效血药浓度的比例从34.78%上升至91.30%。结论：该方法适用于癫痫患者服用拉莫三嗪和奥卡西平后临床常规的TDM，为患者个体化用药提供依据，提高临床疗效、降低不良反应。     

A sensitive, acetonitrile-free, HPLC method for determination of alibendol in dog plasma and its application to pharmacokinetic studies

A sensitive, acetonitrile-free, high performance liquid chromatography method was developed for the quantitation of alibendol concentrations in plasma. To date, only one other method has been reported for the determination of alibendol in plasma. However, that method may lack the sensitivity needed to determine the low plasma levels encountered in pharmacokinetic studies. Following simple extraction, sample components were separated by isocratic reverse-phase chromatography and quantified using UV detection at 315?nm. The mobile phase consisted of a mixture of 0.02?M phosphate buffer and methanol (50:50, v/v), adjusted to pH 3.0 with phosphoric acid. The assay??s lower limit of quantitation (LLOQ) was determined to be 20?ng/ml in a plasma sample volume of 0.5?ml. Sildenafil was used as an internal standard. Observed retention times were approximately 5.0 and 11.5?min for alibendol and sildenafil, respectively. The within-run precision showed RSD values between 5.83 and 16.96?%. The between-run RSD values varied from 6.73 to 17.99?% at the LLOQ. This method was successfully employed to track the concentration of alibendol in beagle dogs following oral administration of the drug. This assay is advantageous as it does not use expensive acetonitrile and is more sensitive than previously reported method.     

A sensitive HPLC method for the determination of fluoxetine and norfluoxetine in human plasma with fluorescence detection

A selective, sensitive, and precise HPLC method for the simultaneous determination of fluoxetine (FL) and its N-demethylated metabolite norfluoxetine (NFL) in human plasma has been developed. Following extraction with n-hexane, FL, NFL, and fluvoxamine (internal standard) were derivatized with 7-chloro-4-nitrobenzofurazan (NBD-Cl) under weakly alkaline conditions. NBD derivatives were extracted with chloroform after acidification and chromatographed on a reversed-phase column with gradient elution using acetonitrile and 0.1 mol/L nitric acid (pH 3) solution. Calibration curves were linear over the range of 1.0-100.0 ng/mL and 0.1-50.0 ng/mL for FL and NFL, respectively, with inter- and intraassay precision given by a relative standard deviation (RSD%) of less than 9.2%. The lower limits of quantification were 1.0 ng/mL for FL and 0.1 ng/mL for NFL. Recoveries of FL and NFL from plasma at three different concentrations were assessed. Average recovery was about 100% for both substances. The assay was applied to pharmacokinetic study in 2 healthy volunteers after a single oral administration of 40 mg of FL.     

Validated HPLC method for determination of amlodipine in human plasma and its application to pharmacokinetic studies

A rapid, simple and sensitive high-performance liquid chromatography (HPLC) method has been developed for quantification of amlodipine in plasma. The assay enables the measurement of amlodipine for therapeutic drug monitoring with a minimum detectable limit of 0.2 ng ml(-1). The method involves simple, one-step extraction procedure and analytical recovery was about 97%. The separation was performed on an analytical 125 x 4.6 mm i.d. Nucleosil C8 column. The wavelength was set at 239 nm. The mobile phase was a mixture of 0.01 M sodium dihydrogen phosphate buffer and acetonitrile (63:37, v/v) adjusted to pH 3.5 at a flow rate of 1.5 ml min(-1). The calibration curve was linear over the concentration range 0.5-16 ng ml(-1). The coefficients of variation for inter-day and intra-day assay were found to be less than 10%.     

A sensitive and simplified HPLC analysis for the determination of fluconazol in human plasma

A sensitive and simplified HPLC assay of fluconazol is described. The calibration curve of fluconazol in plasma ranging 0–10 μ g/ml was linear with the correlation coefficients of 0.9900. The limit of detection was 0.3 μ g/ml. The average recovery of the drug was 89.1±9.05%. After oral administration of single dose(150mg) of fluconazol in man, C_max and T_max were 3 μg/ml and 4hr., respectively.     

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.     

A selective HPLC method for the determination of indapamide in human whole blood: application to a bioequivalence study in Chinese volunteers

Indapamide was extracted from human whole blood with diethyl ether and was determined by a HPLC-UV method using an Inertsil ODS-3 column and an isocratic mobile phase consisting of 55% buffer solution (2 g KH(2)PO(4), 3 ml H(3)PO(4) and 3.5 ml triethylamine in 1l of H(2)O), 40% acetonitrile and 5% methanol for 12.5 min, and then a gradient flush from 100% isocratic to a mixture of 20% isocratic mobile phase and 80% methanol for 3 min. Indapamide and glipizide (internal standard) were eluted from the column at about 10.5 and 12.8 min, respectively. The method had within day precision values in the range +/- 1.2 to +/- 9.7% (n = 5) and between day precision in the range +/- 3.3 to +/- 9.7%. The method was linear over the range of 10-400 ng/ml of indapamide in blood (R = 0.999). The LOQ (s/n = 10) of the method was 10 ng/ml. The method was applied in a study of the pharmacokinetics and bioequivalence of generic indapamide capsules (2.5 mg) in comparison with reference indapamide tablets (2.5mg), in 20 healthy male Chinese volunteers. The mean values of major pharmacokinetic parameters of C(max), AUC(0-48), AUC(0-infinity), T(max), and t(1/2) of indapamide in healthy male Chinese volunteers after po a single 5 mg dose for the test product were 331.0 +/- 39.2 ng/ml, 6,193.7+/-873.5 ng h/ml, 7311.8+/-1,232.3 ng h/ml, 3.2+/-0.9h, and 17.3+/-2.8 h, respectively. There was no significant differences between the two formulations.     

酶放大免疫分析法与高效液相色谱法检测人血浆中丙戊酸浓度比较

目的分析评价酶放大免疫分析法(EMIT)和高效液相色谱法(HPLC)测定人血浆中丙戊酸(VPA)浓度的相关性,为临床监测丙戊酸血药浓度及个体化用药提供参考。方法采用EMIT法和HPLC法同时测定109份血样中丙戊酸的浓度,对测定结果采用双侧配对t检验比较差异,并绘制散点图和Bland-Altman偏差图,考察两种测定结果的相关性。结果以HPLC法测定结果(X_1)与EMIT法测定结果(Y_1)作线性回归方程如下:Y_1=1.016 6X_1+3.082 7(r_1=0.942,n=109),2种检测方法测定丙戊酸血药浓度相关性良好,当样本浓度<100 mg·L~(-1)时,两者差异有显著意义(P=0.024):当样本浓度>100 mg·L~(-1)时,两者差异无显著意义(P=0.596)。Bland-Altman偏差图显示EMIT法测血浆中丙戊酸的浓度较HPLC法偏高。结论 HPLC法和EMIT法测定人血浆中丙戊酸的浓度具有较高的相关性,但均可能存在系统偏差,临床监测丙戊酸的浓度时应予以关注并作相应调整。     

Comparison of plasma and saliva concentrations of lamotrigine in healthy volunteers

The relationship between lamotrigine (CAS 84057-84-1) concentrations in saliva and plasma in healthy volunteers were examined, as well as the possibility of using saliva to monitor levels for effective therapy. The study was performed with 14 healthy volunteers, mean age 23 +/- 2 (SD) years. After single oral doses of 200 mg, plasma and stimulated saliva samples were collected simultaneously at 0, 0.25, 0.5, 1, 2, 4, 6, 8, 10, 12, 24, 48, 72 and 96 h. The pH values of saliva samples were recorded. Lamotrigine concentrations were determined by a validated HPLC method. Fraction of drug bound to plasma proteins was calculated mathematically by the modified Henderson-Hasselbalch equation. Linear regression was used to evaluate the correlations. The remnant of orally administered drug contaminated the saliva samples and gave spuriously high values for up to 2 h, which were omitted. There was significant correlation (r2 = 0.677, p < 0.0001) between plasma and saliva concentrations from 2-96 h after administration. The mean ratio of saliva to plasma concentration was 0.426 +/- 0.153 (mean +/- SD). Protein binding, calculated from the concentrations in saliva was 57.5 +/-15.1% (mean +/- SD). Noncompartmental analysis was conducted with the program Kinetica. Plasma t1/2 and MRT were not significantly different from those found from saliva. The mean values of lamotrigine peak saliva concentrations (C(max)), areas under the curve of concentration versus time from zero to infinity (AUC(0-->infinity)), and areas under the curves of the product of time and concentration versus time from zero to infinity (AUMC(0-->infinity) were proportionally lower than in plasma. The results support the use of saliva concentration as a convenient, painless and noninvasive alternative to plasma for monitoring lamotrigine therapy.     

A sensitive LC-ESI-MS method for the determination of indapamide in human plasma: method and clinical applications

A sensitive LC-ESI-MS method for the determination of indapamide in human plasma using glibenclamide as the internal standard (IS) was established. Following acidification with 1 M hydrochloric acid solution, plasma samples were extracted with ethyl acetate and separated on a C(18) column with a mobile phase of 10 mM ammonium acetate-methanol (22:78, v/v). Indapamide was determined using electrospray ionization in a single quadrupole mass spectrometer. LC-ESI-MS was performed in the selected ion monitoring (SIM) mode using target ions at m/z 364.3 for indapamide and m/z 492.4 for the IS. Calibration curves were linear over the ranges of 0.1-100 ng/ml for indapamide. The lower limit of quantification was 0.1 ng/ml. The intra- and inter-assay precisions were less than 9.5% and 10.6%, respectively. The mean plasma extraction recovery of indapamide was 90.5-93.9%. The method has been successfully applied to study the pharmacokinetics of indapamide in healthy male Chinese volunteers.     

Determination of 5-fluorouracil in human plasma by a simple and sensitive reversed-phase HPLC method

A simple and sensitive reversed-phase HPLC method with UV detection was developed and validated for the quantitation of 5-fluorouracil (5-FU) in human plasma. After acidification and salting out, 5-FU was extracted into ethyl acetate and back-extracted into a basic buffer. The extract was adjusted to neutral pH before being injected onto the HPLC column. 5-FU was separated from the matrix components on a YMC ODS-AQ column at 40°C using an aqueous mobile phase of 10 mM potassium phosphate at pH 5.5. A linear gradient of 0–25% methanol wash eluted late peaks, maintained column performance, and increased column stability. The run time was 20 min. The linear range was 25–300 ng ml⁻¹ (r²>0.999). The limit of quantitation was 25 ng ml⁻¹, with a signal-to-noise ratio of 23:1. Interday precision and accuracy of quality control samples were 6.2–8.4%, relative standard deviation and −0.1– + 1.9% relative error.