HPLC法快速测定人血浆中酒石酸美托洛尔的浓度

目的:建立快速测定人血浆中酒石酸美托洛尔浓度的方法。方法:血浆样品以3-叔丁基甲醚处理后,采用高效液相色谱法进样测定,色谱柱为Kromasil LC-18DB,流动相为乙腈-10 mmol/L辛烷磺酸钠溶液(p H=2.0)(30∶70),柱温为25℃,流速为1.3ml/min,荧光检测波长为267 nm(激发波长)、290 nm(发射波长),内标为吲哚洛尔。结果:酒石酸美托洛尔血药浓度在2~300 ng/ml范围内线性关系良好(r=0.999 9),最低检测浓度为1 ng/ml。平均方法回收率为(101.13±4.0)%,日内RSD≤2.47%,日间RSD≤4.51%。结论:该方法简便、快速、灵敏、重现性好,适用于酒石酸美托洛尔临床血药浓度监测及人体药动学研究。     

大鼠血浆中毛萼乙素的HPLC测定

建立了HPLC法测定大鼠血浆中的毛萼乙素.采用液相萃取法处理血浆样品,色谱柱为C_(18)柱,流动相为乙腈-水(43:57),检测波长233nm.毛萼乙素在40～8 000 ng/ml浓度范围内线性关系良好.方法回收率101.6%～106.5%,日内、日间RSD为4.3%～12.4%.     

高效液相色谱荧光法测定Beagle犬血浆中酒石酸美托洛尔对映体浓度

目的:建立Beagle犬血浆中酒石酸美托洛尔对映体浓度的高效液相色谱荧光测定方法。方法:Beagle犬口服消旋体酒石酸美托洛尔100mg后取1·5h血样进样测定,手性色谱柱为ChiralcelOD-H,流动相为正己烷-异丙醇-二乙胺(65∶35∶0·1),荧光检测波长Ex和Em分别为267nm、290nm,柱温为25℃,流速为0·6ml/min,内标物为(S)-阿替洛尔。结果:酒石酸美托洛尔2种对映体检测浓度在10~2000ng/ml范围内线性关系良好,平均回收率为96·12%~116·9%(RSD<7·6%)。结论:该分析方法选择性好、准确性高、重现性好,适于酒石酸美托洛尔对映体的药动学研究。     

人血浆中西酞普兰的HPLC-荧光法测定

建立了HPLC-荧光法测定人血浆中西酞普兰的浓度.血浆样品经液-液萃取后测定,用Zorbax SB C8色谱柱,磷酸二氢铵缓冲液(pH 3.5)-乙腈(65:35)为流动相,激发波长240 nm,发射波长302 nm.血浆中西酞普兰线性范围为1～100ng/ml,最低定量浓度为1ng/ml,日内、日间RSD均小于4.0%,方法平均回收率为93.2%.     

中空纤维膜液相微萃取-HPLC法测定复方穿心莲片中脱水穿心莲内酯含量

目的:建立一种新方法—中空纤维膜液相微萃取-高效液相色谱法测定复方穿心莲片中脱水穿心莲内酯的含量。法:将样品用甲醇浸泡、超声、过滤后,配成样品溶液。经用装有正辛醇的中空纤维膜进行液相微萃取后,用 HPLC 法测定复穿心莲片中脱水穿心莲内酯的含量,色谱柱为 Waters symmetry C_(18)柱(150 mm×3.9 mm,5μm),流动相为甲醇-水(60:40结果:脱水穿心莲内酯浓度范围在100～8000 ng·mL~(-1)内线性关系良好,其加样回收率>96.0%,RSD<3.8%。结论:通液相微萃取后,能有效地去除检测复方穿心莲片中的干扰物质,样品处理无需经过中性氧化铝柱纯化,获得了较高的选择性该方法简便、快速,与不经过液相微萃取的液相色谱方法相比,灵敏度提高约128倍,是测定复方穿心莲片中脱水穿心莲内含量的一种好方法。     

固相萃取结合高效液相色谱法测定人血浆中酒石酸唑吡坦的浓度

目的:建立固相萃取结合高效液相色谱法测定人血浆中酒石酸唑吡坦浓度。方法:血样经固相萃取法提取,采用大连依利特色谱柱(Hypersil BDS C₁₈),流动相:甲醇-乙腈-水（45∶45∶110）,流速1.0ml·min^-1,柱温:35℃;检测波长:254 nm,内标法定量。结果:血浆中酒石酸唑吡坦浓度在0.511 5⁴.092 0μg·ml^-1线性关系良好,相对回收率大于90%,精密度日间、日内RSD均小于10%。结论:该测定方法简单、快速、准确、干扰小,适用于临床测定酒石酸唑吡坦药物浓度。     

高效液相色谱法测定人血浆中硝苯地平的浓度

目的:建立高效液相色谱法测定人血浆中硝苯地平的浓度.方法:分析柱为Micro Pah C 18分析柱.流动相为甲醇:水:冰乙酸:正丁胺(77:23:0.02:O.01).检测波长为235nm.采取乙醚二次萃取法提取血浆中的硝苯地平及内标安定.结果:测定硝苯地平的日内及日间精密度的RSD均小于10%.血药浓度在6.16～197.20 ng/ml范围内线性关系良好(r=0.9994).最低检测限为2ng/ml.结论:本方法适用于硝苯地平的药物动力学研究.     

阳离子色谱柱-HPLC法测定人血浆中氯氮平及去甲氯氮平浓度

目的:建立同时测定人血浆中氯氮平及去甲氯氮平浓度的阳离子色谱柱-HPLC法。方法:用强阳离子交换色谱柱-HPLC测定人血浆中氯氮平及去甲氯氮平浓度,色谱柱为强阳离子交换色谱柱(4.6 mm×100 mm,5μm),流动相为乙腈-0.01 mol·L-1磷酸铵溶液(pH 5.1)(60∶40),流速为1.2 mL·min-1,紫外检测波长为257 nm,以AF2672为内标,含氯氮平及去甲氯氮平的血样经C8固相小柱萃取后进样。结果:阳离子色谱柱-HPLC测得氯氮平、去甲氯氮平的色谱峰峰形对称、无内外源性物质干扰,检测结果稳定、可靠、重复性好;通过改变流动相pH易于调整各化合物保留时间,同时去除空白血浆干扰;固相萃取前处理简单、省时且萃取回收率较高。氯氮平及去甲氯氮平浓度在50.4～990 ng·mL-1范围内线性关系良好;氯氮平及去甲氯氮平低(50.4 ng·mL-1)、中(396 ng·mL-1)、高(990 ng·mL-1)3个浓度的萃取回收率均大于91%,日内和日间RSD均小于4%(n=5)。结论:阳离子色谱柱-HPLC法测定人血浆中氯氮平及去甲氯氮平浓度,色谱峰峰形对称,保留时间适宜且无杂质干扰,适用于临床氯氮平及去甲氯氮平血药浓度监测和药动学研究。     

高效液相色谱法测定人血浆中酒石酸唑吡坦的浓度

目的:建立HPLC法测定人血浆中酒石酸唑吡坦浓度。方法:采用Hypersil C_(18)色谱柱(200 mm×4.6 mm,5μm);流动相:0.02 mol·L~(-1)磷酸二氢钾(含0.5%三乙胺,pH=4.0)-甲醇(38:62);流速:1.2 ml·min~(-1);柱温:30℃;激发波长:300 nm,发射波长:385 nm。结果:血浆酒石酸唑吡坦在2～200 ng·ml~(-1)浓度范围内线性关系良好(r=0.999 9)。低、中、高3种浓度相对回收率分别为102.0%,102.1%,98.9%,日内、日间RSD均小于10%。结论:本法可用于血浆中酒石酸唑吡坦浓度的测定。     

在线固相萃取-HPLC法测定比格尔犬血浆中丹参素的含量

目的:建立测定比格尔犬血浆中丹参素含量的在线固相萃取-HPLC法。方法:血浆样品经蛋白质沉淀,采用Lichrospher C18为富集柱,以乙腈-10 mmol/L NaH2 PO4(5∶95)为富集流动相,流速2 ml/min;选择Ultimate XB-C18(50 mm×4.6 mm,5μm)为分析柱,以乙腈-10 mmol/L NaH2 PO4(11∶89)为分析流动相,流速为0.8 ml/min,检测波长285 nm。结果:丹参素在20~2 000 ng/ml浓度范围内线性关系良好,最低定量限为20.0 ng/ml,日内和日间RSD均<10%,平均绝对回收率>80%。结论:本方法简便、快速、灵敏、可靠,适用于比格尔犬血浆中丹参素含量的测定。     

Hollow fiber-based liquid phase microextraction (HF-LPME) as a new approach for the HPLC determination of fluoroquinolones in biological and environmental matrices

In this paper, a three phase hollow fiber-based liquid phase microextraction (HF-LPME) combined with a HPLC procedure using diode array (DAD) and fluorescence detection (FLD) has been developed for the determination of eight widely used fluoroquinolones: marbofloxacin (MRB), norfloxacin (NRF), ciprofloxacin (CPR), danofloxacin (DNF), enrofloxacin (ENR), gatifloxacin (GTF), grepafloxacin (GRP) and flumequine (FLM). A Q3/2 Accurel PP polypropylene hollow fiber supporting 1-octanol was used between a 2 M Na2SO4 aqueous solution (pH 7) as donor phase and aqueous solution (pH 12) as acceptor phase. The microextraction parameters were optimised from an experimental central composite design. The procedure allows very low detection and quantitation limits of 0.3-16 ng L(-1) and 1-50 ng L(-1), respectively. The proposed method was applied to the determination of the analytes in bovine urine and in environmental water samples (surface, tap and wastewater).     

Determination of metoprolol enantiomers in human urine by coupled achiral-chiral chromatography

Achiral chiral column switching HPLC assay was developed to allow the separation and quantitation of the enantiomers of metoprolol in human urine by means of fluorescence detection. Urine samples were prepared by liquid liquid extraction, followed by HPLC. The racemic metoprolol and internal standard were separated from the interfering components in urine and quantified on the silica column, and the enantiomers were determined on a Chiralcel OD chiral stationary phase. The two columns were connected by a switching valve equipped with a silica trap column. Detection limit was 25 ng/ml for each enantiomer. The intra-day variation ranged between 0.38 and 4.94% in relation to the measured concentration and the inter-day variation was 0.15-3.13%. It has been applied to the determination of (R)-(+)-metoprolol and (S)-(-)-metoprolol in urine from healthy volunteers dosed with racemic metoprolol tartrate.     

Hollow fiber liquid-phase microextraction for the determination of nimesulide in human plasma and its application to a pharmacokinetic study

A hollow fiber liquid-phase microextraction (HF-LPME) method in combination with HPLC-UV for the determination of nimesulide in human plasma was developed and validated. A small volume of dihexyl ether contained within a polypropylene hollow fiber was used for the extraction of nimesulide from acidified plasma solutions. Factors affecting the extraction efficiency were optimized and discussed. With HPLC-UV as the end analysis technique, the procedure was validated for nimesulide in the concentration range of 50-5000 ng/mL. The intra- and inter-assay precisions were less than 9.1%, and accuracy was within 3.2%. The lower limit of quantification (LLOQ) was 50 ng/mL. Enrichment factor from 144-fold to 156-fold was achieved at three quality control (QC) concentrations. The mean extraction recovery was greater than 41.2%. This method was successfully applied for the evaluation of pharmacokinetics of nimesulide after single oral doses of 100 mg nimesulide to six healthy Chinese volunteers.     

高效液相色谱法测定复方美托洛尔胶囊中酒石酸美托洛尔的含量

目的建立高效液相色谱法测定复方美托洛尔胶囊中酒石酸美托洛尔的含量。方法采用美国Agilent高效液相色谱仪,以醋酸盐缓冲液-乙腈(850∶150)为流动相,流速2mL.min-1,柱温30℃,检测波长为275nm。结果美托洛尔在100.6~603.6μg.mL-1范围内,线性关系良好,r=0.9996。回收率99.2%。结论该法可准确测定复方美托洛尔胶囊中酒石酸美托洛尔的含量,适用于产品的质量控制。     

HPLC法测定酒石酸美托洛尔片的含量和有关物质

目的：建立高效液相色谱法测定酒石酸美托洛尔片的含量和有关物质。方法：色谱柱：Eclipse XDB—C18柱（150mm×4．6mm,5μm）;流动相：甲醇-水（1：1）（1000ml,加庚烷磺酸钠960mg与无水醋酸钠82mg使溶解,加冰醋酸0．60ml,混匀;检测波长：224nm;流速：1．0ml·min^-1。结果：酒石酸美托洛尔浓度在0．04—0．32mg·ml^-1范围内,线性关系良好,回归方程Y=31557X＋94．1（r=0．9999）。平均回收率为99．6％（n=9）,RSD为0．3％。结论：所用方法快速、简便、准确,专属性强。     

Extraction and determination of trace amounts of chlorpromazine in biological fluids using hollow fiber liquid phase microextraction followed by high-performance liquid chromatography

In the present work, hollow fiber liquid phase microextraction (HF-LPME) in conjunction with reversed-phase HPLC/UV was developed for extraction and determination of trace amounts of chlorpromazine in biological fluids. The drug was extracted from an 11 ml aqueous sample (source phase; SP) into an organic phase impregnated in the pores of the hollow fiber (membrane phase; MP) followed by the back-extraction into a second aqueous solution (receiving phase; RP) located in the lumen of the hollow fiber. The effects of several factors such as the nature of organic solvent, compositions of SP and RP solutions, extraction time, ionic strength and stirring rate on the extraction efficiency of the drug were examined and optimized. Under the optimal conditions, enrichment factor of 250, dynamic linear range of 1–500 μg l⁻¹, and limit of detection of 0.5 μg l⁻¹ were obtained for the drug. The percent relative intra-day and inter-day standard deviation (R.S.D.%) based on three replicate determinations were 6.7 and 10.3%, respectively. The method was applied to drug level monitoring in the biological fluids and satisfactory results were obtained.     

非水液相微萃取-高效液相色谱法测定大黄中5种游离蒽醌类化合物

目的利用非水液相微萃取-高效液相色谱法(HPLC)同时测定大黄中游离蒽醌类化合物的含量。方法利用自制的液相微萃取装置,以聚偏氟乙烯中空纤维为溶剂载体,正己醇为萃取溶剂,供相为甲醇,接受相为1mmol/LNaOH,搅拌速度为1500r/min,萃取时间为50min。萃取结束,接受相在434nm处进行HPLC分析。结果在优化的液相微萃取条件下,芦荟大黄素、大黄酸、大黄素、大黄酚和大黄素甲醚的线性范围、回收率、精密度和检测限分别为:0.015～13.2、0.075～11.2、0.085～12.8、0.096～14.4、0.117～17.6μg/ml;103.8%～106.1%;2.5%～5.2%和2.9～20.0ng/ml。结论本法选择性高,有机溶剂消耗少,能快速、准确地测定大黄药材中游离蒽醌类化合物。     

HPLC法测定酒石酸美托洛尔注射液的含量和有关物质

目的：建立HPLC法测定酒石酸美托洛尔注射液的含量和有关物质。方法：色谱柱：phenomenex luna C18柱（150mm×4．6mm,5μm）,流动相：甲醇-庚烷磺酸钠缓冲溶液（水450ml,加庚烷磺酸钠960mg,加无水醋酸钠82mg,加冰醋酸0．57m1）（49：51）,流速：1．0ml·min^-1,检测波长：275nm。结果：在选定的色谱条件下,主药与杂质分离良好。酒石酸关托洛尔在28．94～964．80μg·m^-1范围内线性关系良好（r=0．9999,n=6）。结论：该方法可用于酒石酸关托洛尔注射液的含量和有关物质的测定。     

Three-phase,liquid-phase microextraction combined with high performance liquid chromatography-fluorescence detection for the simultaneous determination of fluoxetine and norfluoxetine in human plasma

A three-phase, liquid-phase microextraction using a hollow fibre (HF-LPME) combined with high performance liquid chromatography-fluorescence detection (HPLC-FL) was developed for the analysis of fluoxetine (FLX) and its active metabolite, norfluoxetine (NFLX), in human plasma. An HF-LPME system using a disposable 7-cm polypropylene porous hollow fibre, 5 mL of alkaline plasma solution (donor phase), n-hexyl ether (extraction solvent) and 20 mM hydrochloric acid (acceptor phase) was used in the extraction. The method was validated after optimisation of several parameters that influence LPME efficiency. A reverse-phase LiChrospher 60 RP-Select B column (125 mm × 4 mm, 5 μm particle size) was used with 0.005 M sodium acetate buffer (pH 4.5) and acetonitrile at a 50:50 (v/v) as the mobile phase at a flow rate of 0.6 mL min⁻¹. In these conditions satisfactory chromatographic resolution and efficiency for the analytes were obtained. Fluorescence detection at 230 nm excitation wavelength and 290 nm emission wavelength was performed. Linearity over a range of 5–500 ng mL⁻¹, with determination coefficients (R²) of 0.9999 and 0.9962 for FLX and NFLX, respectively, was established. Venlafaxine was used as the internal standard for both analytes. Extraction recoveries from plasma samples were 70.9% for FLX and 59.7% for NFLX. The intra-day coefficients of variation (CVs) were below 5.4%, and inter-day CVs were below 13.0%, for both analytes at concentrations of 20, 80 and 160 ng mL⁻¹. HF-LPME extraction followed by HPLC-FL detection for FLX and NFLX analyses demonstrated excellent sample clean-up and selectivity. This method was simple, cheap, and easy to perform, yielding substantial analytes enrichment. The method was applied to the analysis of samples from 12 patients under fluoxetine treatment and proved suitable for routine therapeutic drug monitoring for this antidepressant.