RP-HPLC法同时测定藏药珠芽蓼中牡荆素、槲皮苷和槲皮素的含量(英文)

采用RP-HPLC法建立了同时测定珠芽蓼中牡荆素、槲皮苷、槲皮素三种黄酮的方法,色谱柱为kromasilC18(250 mm×4.6 mm,5μm),以甲醇-水(含0.25%磷酸)为流动相梯度洗脱,流速1 mL/min,检测波长360 nm。结果表明,牡荆素、槲皮苷和槲皮素质量浓度在8～40、5～50μg/mL和5.33～52μg/mL范围内与色谱峰面积均呈良好的线形关系,平均回收率分别为100.3%、98.9%和100.4%,日内稳定性偏差分别为0.78%,0.51%和0.38%,日间稳定性偏差为2.6%,1.9%和2.1%。该方法简单、准确,可为珠芽蓼的质量控制提供科学依据。     

反相高效液相色谱法测定升麻药材中升麻苷H-1的含量

建立升麻药材中升麻苷H-1含量测定的反相高效液相色谱分析方法。使用H&E XP ODS-A色谱柱(4.6 mm×250 mm5,μm),流动相为乙腈-水-磷酸(35∶65∶0.4),流速为1 mL/min,柱温为26℃,检测波长203nm。测得升麻苷H-1线性范围为0.95～28.5μg/mL(r=0.9999),平均回收率(n=6)为99.35%(RSD=2.79%),不同来源批次升麻药材含量测定结果表明,升麻药材中升麻苷H-1含量为0.53%～1.09%,综合分析批样品含量测定结果,建议升麻药材中含升麻苷H-1应不低于0.4%。本方法简便、准确、重复性好、专属性强,可作为升麻药材质量控制方法。     

高效液相色谱法测定罗汉果根中两个降三萜配糖体

建立了高效液相色谱测定罗汉果根中两个结构新颖的降三萜糖苷,罗汉果酸苷乙Ⅱ(Siraitic acidⅡB)和罗汉果酸苷甲Ⅱ(Siraitic acidⅡA)含量的方法。色谱条件:色谱柱:ZORBAX SB-C18柱(4.6mm×150mm,5μm);柱温:25℃;流动相:乙腈-水(梯度洗脱:0→4min:35%乙腈;4→16min:35%→45%乙腈);流速:0.5mL/min;检测波长:230nm;进样量:10μL。结果表明,罗汉果酸苷乙Ⅱ在0.05～0.3mg/mL,罗汉果酸苷甲Ⅱ在0.05～0.25mg/mL之间线性关系良好。该方法稳定,精密度、重现性好,且样品处理简单,提取与检测时间短。     

刺五加及短梗五加根皮有效成分含量对比研究

目的:建立刺五加及短梗五加根皮刺五加苷B、刺五加苷E及异嗪皮啶的高效液相测定方法,测定刺五加及短梗五加中上述成分的含量,对比研究刺五加及短梗五加根中的成分含量差异。方法:采用高效液相色谱法,Agilent TC-C18色谱柱(150 mm×4.6 mm,5μm),以甲醇(A)-水(B)为流动相,梯度洗脱:(0～15 min:20A;15～20 min:21A;20～35 min:30A),流速0.8 mL.min-1,柱温30℃,检测波长210 nm。结果:样品进样量在各自浓度范围内呈良好的线性关系,加样回收率分别为99.1%、97.3%和99.9%,RSD为0.7%和1.3%和0.6%。结论:本法操作简便,分离效果好,可用于刺五加及短梗五加根皮刺五加苷B、刺五加苷E及异嗪皮啶的含量测定。     

蒙药材刺柏叶中槲皮苷的含量测定

本实验建立了蒙药材刺柏叶中槲皮苷的HPLC含量测定方法.色谱条件为:Diamonsil C18(4.6 mm ×250 mm,5μm)和Shim-Pack C18(4.6 mm× 250 mm,5μm),流动相:甲醇-0.01 mol/L磷酸二氢钾溶液-冰醋酸(40∶ 60∶ 1.5),检测波长:254 nm,流速:1.0 mL/min,柱温:30℃.槲皮苷进样量在40.2 ng ～603.0 ng范围内线性关系良好(r =0.9998),其平均加样回收率为100.45％,RSD为0.69％(n=6).本方法简便易行,结果准确,重复性好,可用于刺柏叶中槲皮苷的含量测定.     

HPLC法测定18种菊科植物中总黄酮醇苷含量研究

目的:建立测定青海产18种菊科植物中总黄酮醇苷含量的方法。方法:Reprosil C18-AQ色谱柱(4.6 mm×250 mm,5μm),流动相甲醇-0.2%甲酸溶液(48∶52),检测波长360 nm,流速1.0 mL/min,柱温40℃,进样量10μL。结果:槲皮素、山柰素和异鼠李素3种黄酮类化合物在1.06～212、1.06～212、0.93～186μg/mL浓度范围内均线性关系良好。本方法精密度、稳定性和重复性良好。结论:该方法准确稳定,重复性好,可用于菊科植物的质量控制。     

HPLC法测定前列舒乐颗粒中淫羊藿苷的含量

目的:建立前列舒乐颗粒中淫羊藿苷的HPLC测定方法。方法:以C18(4.6 mm×15 mm,5μm)为色谱柱,以乙腈-水(30∶70)为流动相,流速1 mL/min,测定波长270 nm。结果:线性范围在0.40～4.00μg(r=0.9999),平均回收率为99.79%,RSD=2.5%(n=6)。结论:本方法简便可行,重现性好,结果可靠,可以用来测定淫羊藿苷的含量,控制前列舒乐颗粒的质量。     

暗褐网柄牛肝菌尿苷和麦角甾醇含量测定及 指纹图谱分析

采用HPLC测定暗褐网柄牛肝菌(Phlebopus portentosus)中尿苷和麦角甾醇含量并建立其指纹图谱。结果表明,最佳分析条件为Waters HSST3色谱柱(4.6 mm×150 mm,5μm),流动相为0.1%甲酸水(A)-甲醇(B),梯度洗脱(0～5 min,0%B; 5～15 min,0→4%B; 15～45 min,4%→100%B; 45～60 min,100%B),流速1 mL/min,检测波长260 nm,柱温30℃,进样量0. 5μL。尿苷和麦角甾醇分别在质量浓度0. 003 4～0.34 mg/mL和0. 060 5～1. 21 mg/mL时线性关系良好;平均加样回收率分别为98. 31%(RSD 2. 98%)和102. 72%(RSD 2.84%)。含量测定结果显示,10批样品尿苷和麦角甾醇的含量范围分别为0～2.00 mg/g和3.38～9. 10 mg/g。建立了10批暗褐网柄牛肝菌的共有峰模式,标记了6个共有峰,10批样品的指纹图谱相似度均0.95。方法学考察结果表明本实验建立的HPLC含量测定和指纹图谱分析方法准确可靠,可用于暗褐网柄牛肝菌的质量评价。     

HPLC法同时测定藏药岷县龙胆中4种有效成分的含量

采用高效液相色谱法同时测定藏药岷县龙胆中獐牙菜苦苷、龙胆苦苷、獐牙菜苷和异荭草苷含量。结果测定岷县龙胆中獐牙菜苦苷、龙胆苦苷、獐牙菜苷和异荭草苷的含量分别为0.108%、1.46%、2.21%、1.61%。色谱条件为:色谱柱:XDB-C18(4.6 mm×250 mm,5μm);柱温:25℃;流速:1.0 mL/min;流动相:甲醇/0.01%乙酸水溶液梯度洗脱;检测波长260 nm。本方法快速、准确、重复性好,适用于岷县龙胆中4种有效成分的含量测定,可为藏药岷县龙胆药材的质量控制提供依据。     

HPLC-ELSD法测定消栓通颗粒中黄芪甲苷的含量

目的:建立测定消栓通颗粒中黄芪甲苷含量的新方法.方法:采用高效液相色谱-蒸发光散射检测法测定,色谱柱为kromasilC18柱,以乙腈-水(32:68)为流动相,流速为1.0ml/min,ELSD条件为飘逸管温度为105℃,载气流速为2.5L/min;测定消栓通颗粒中黄芪甲苷的含量.结果:黄芪甲苷在1.08μg-6.48μg范围内与峰面积线性关系良好(r=0.9999),平均加样回收率为100.2%,RSD为0.93%.结论:方法灵敏、可靠、准确、重复性好,可作为消栓通颗粒的质量控制方法.     

HPLC测定7种龙胆科植物花中龙胆苦苷与獐牙菜苦苷的含量

目的:建立7种龙胆花中龙胆苦苷和獐牙菜苦苷含量测定的HPLC方法。方法:采用微波辅助动态回流法进行提取,色谱条件:Fusion-RP 80 A C18柱(150 mm×4.6 mm,5μm);流动相:甲醇-0.2%磷酸溶液梯度洗脱(0~25 min:15%~30%);流速:1 mL/min;柱温:30℃;检测波长240 nm。结果:7种龙胆花中獐牙菜苦苷和龙胆苦苷的色谱峰与共存组分完全达到基线分离,线性范围分别为0.105~0.945μg(r=0.999 9),0.3~0.7μg(r=0.999 9),平均加样回收率分别为97.8%(RSD=1.02%),98.9%(RSD=1.51%)。结论:所建立的方法测定快速,结果准确可靠。     

发酵法生产紫杉醇的检测分析

目的:采用高效液相色谱法对发酵液中的紫杉醇进行测定。方法:将紫杉醇产生菌发酵产物经乙酸乙酯萃取得测定样品,高效液相色谱分析方法为苯基柱(250mm×4.6mm,5μm),流动相乙腈-甲醇-水(36∶4∶60),流速1mL/min,检测波长227nm,进样体积20μL,柱温室温。结果:紫杉醇与干扰物可达到基线分离,在2.2～110μg/mL范围内,紫杉醇的峰面积与浓度线性关系良好,相关系数0.9996,平均回收率为99.55%,RSD为0.60%。结论:与使用C18柱色谱条件相比,该分析方法灵敏度高,不需要复杂的样品前处理过程,仪器配置不复杂、操作方便、准确性高,可有效地检测发酵液中紫杉醇的含量。     

Determination of isoflavones in soybean food and human urine using liquid chromatography with electrochemical detection

A highly sensitive high-performance liquid chromatographic method with electrochemical detection (HPLC-ED) was developed for the determination of isoflavones. Electrochemical behaviour of daidzein and genistein was studied on carbon paste electrode (CPE) by adsorptive transfer stripping square wave voltammetry. The obtained electrochemical results were used for the development of HPLC-ED method. Furthermore, isoflavones were separated on an Atlantis dC18 column using a mobile phase consisting of acetonitrile (solvent A) and 0.15M acetate buffer of pH 5.5 (solvent B) at a flow rate 0.4 mL/min. A linear gradient profile (solvent B) was at 0-2 min 87%; 22 min 60%; 27 min 50%; 31 min 45%; 47 min 87%. Full scan of multi-channel coulometric detection was tested and optimal potential at 450 mV was chosen for our purposes. Calibration curves were linear (daidzein R(2) = 0.9993 and genistein R(2) = 0.9987). The detection limit for daidzein/genistein was 480/394 pg/mL (1.8/1.5 nM) and per column 2.4/1.9 pg. Isoflavones extracted from soybean products (farina, meat, milk) by the accelerated solvent extraction (ASE) procedure and isoflavones present in human urine were determined by the HPLC-ED method.     

The tissue distribution in mice and pharmacokinetics in rabbits of oxaliplatin liposome

A rapid, sensitive, and simple high-performance liquid chromatographic (HPLC) method with an ultraviolet detector (UV) has been developed for the determination of oxaliplatin in the plasma of rabbits and tissues of mice. The sample preparation was carried out by complexation with 0.5?mL of DETC (diethyl-dithiocarbamate) solution and extracted by ether and chloroform. Then, 20?μL of supernatant was injected into the HPLC system with 0.25?mol/L of sodium chloride solution and methanol (30:70 v/v) as the mobile phase at a flow rate of 1.0?mL/min. Separation was performed with a C₁₈ column at 25°C. The peak was detected at 254?nm. The calibration curve was linear (R²?≥?0.9995) in the concentration range of 0.1~200?μg/mL in plasma and tissues. The intra- and interday variation coefficients were not more than 2.61 and 3.83%, respectively. The limit of detection was 20?ng/mL. The mean recoveries of oxaliplatin were ranged from 97.83 to 104.17% in plasma and tissues. The present method has been successfully applied to the pharmacokinetic study of oxaliplatin liposome in mice and rabbits.     

蛹虫草子实体中海藻糖含量的测定

为分析食用菌中海藻糖的含量,以蛹虫草子实体为材料,比较了提取溶剂、提取方式及提取时间等条件对海藻糖提取效果的影响,确定海藻糖分析的前处理方法为：1g子实体粉中加入100mL 90%乙醇热回流提取1h。采用高效液相色谱法测定海藻糖,优化后的色谱条件为：SUGAR SP0810柱（300mm×8mm）,超纯水洗脱,流速0.5mL/min,柱温70℃,示差折光检测器检测,进样量10μL。方法学考察结果表明,该方法准确度高,稳定性、精密度、重现性好,对海藻糖标准品的检测特异性良好,适用于蛹虫草子实体中海藻糖含量的     

HPLC-UV method development for fentanyl determination in rat plasma and its application to elucidate pharmacokinetic behavior after i.p. administration to rats

A simple, rapid and validated high performance liquid chromatography method with UV detection for the quantification of an opioid agonist, fentanyl (FEN), in rat plasma was developed. The assay procedure involved chromatographic separation using a ZIC-HILIC SeQUANT column (250 mm × 4.6 mm, i.d., 5 μm) and a mobile phase of acetonitrile and acetate buffer (pH 3.4, 20mM) of ratio (=65:35, v/v) at a flow rate of 1.2 mL/min and detection wavelength of 201 nm. Plasma sample (100 μL) pretreatment was based on simple deprotienization by acetonitrile spiked with clonidine as an internal standard (I.S.) of 20 ng/mL followed by extraction with tert-butyl methyl ether and centrifugation. The organic layer was evaporated under N(2) gas and reconstituted with 100 μL of acetate buffer (pH 3.4, 20mM), and 50-μL portions of reconstituted sample were injected onto the column. Sample analysis including sample pretreatment was achieved within 35 min. Calibration curve was linear (r ≥ 0.998) from 5 to 100 ng/mL. Both intra- and inter-day assay precisions that are presented through RSD were lower than 12.6% for intra-day and lower than 12.0% for inter-day assessment. Limit of detection was 0.8 ng/mL at S/N of 3. This method was omitting the use of expensive solid phase extraction and time consuming liquid extraction procedures. Moreover, the present method was successfully applied to study pharmacokinetic parameters of FEN after intraperitoneal administration to male Wistar rat. Pharmacokinetic parameters estimated by using moment analysis were T(1/2) 198.3 ± 44.7 min, T(max) 28.3 ± 2.9 min and AUC(0-180) 15.6 ± 2.9(× 10(2))ngmin/mL.     

Quantification of CLR1401, a novel alkylphosphocholine anticancer agent,in rat plasma by hydrophilic interaction liquid chromatography–tandem mass spectrometric detection

A rapid and specific LC–MS/MS based bioanalytical method was developed and validated for the determination of 18-(p-iodophenyl)octadecyl phosphocholine (CLR1401), a novel phosphocholine drug candidate, in rat plasma. The optimal chromatographic behavior of CLR1401 was achieved on a Kromasil silica column (50 mm × 3 mm, 5 μm) under hydrophilic interaction chromatography. The total LC analysis time per injection was 2.8 min with a flow rate of 1.5 mL/min under gradient elution. Liquid–liquid extraction in a 96-well format using ethyl acetate was developed and applied for method validation and sample analysis. The method validation was conducted over the curve range of 2.00–1000 ng/mL using 0.0500 mL of plasma sample. The intra- and inter-day precision and accuracy of the quality control samples at low, medium, and high concentration levels showed ≤ 5.9% relative standard deviation (RSD) and −10.8 to −1.4% relative error (RE). The method was successfully applied to determine the toxicokinetics of CLR1401 in rats from three dose groups of 0.4, 4.0, and 10.0 mg/kg/day via intravenous administration.     

昆虫体内多胺的高效液相色谱(HPLC)测定

建立丹磺酰氯柱前衍生HPLC快速测定昆虫体内多胺含量的方法。以C18(250mm×4.6mm,5μm)为固定相,甲醇和水为流动相,梯度洗脱,柱温40℃,流速1mL/min,荧光检测波长激发波长(Ex)280nm,发射波长(Em)515nm,测得腐胺(put)、亚精胺(spd)和精胺(spm)三者回收率分别为98.7%,99.2%和97.8%,回归方程线性良好(r值均大于0.99),分析时间为16min。该法简洁、快速、灵敏度高、重现性好,可有效分析昆虫及其他生物样品中微量多胺的含量。     

高效液相色谱法测定山楂叶中熊果酸含量的研究

本文建立了一种可靠性高、重现性好的高效液相色谱(HPLC)测定山楂叶中熊果酸含量的方法,在测定中采用富集和固相萃取组合纯化工艺去除干扰物质。高效液相色谱测定条件为Hypersil(ODS)色谱柱,流动相为甲醇:0.2%磷酸二氢钠(90∶10,V/V),检测波长210nm,流速0.8mL/min。熊果酸浓度在100~800μg/mL与峰面积存在良好线性关系(r2=0.9992),该方法准确可靠,日内稳定性标准偏差在0.6%~1.5%,日间稳定性标准偏差在0.7%~2.6%。为不同产地山楂叶中熊果酸含量建立有效的分析方法。