Determination of active ingredients of Ilex Purpurea Hassk and its medicinal preparations by capillary electrophoresis with electrochemical detection

A method based on capillary electrophoresis with electrochemical detection has been developed for the separation and determination of isovanillic acid, gentisic acid, kaempferol, quercetin, caffeic acid and protocatechuic acid in Ilex Purpurea Hassk and its medicinal preparations for the first time. The effects of working electrode potential, pH and concentration of running buffer, separation voltage and injection time on CE-ED were investigated. Under the optimum conditions, the analytes could be separated in a 50 mmoll(-1) borate buffer (pH 9.0) within 21 min. A 300 microm diameter carbon disk electrode has a good response at +0.95 V (versus SCE) for all analytes. The response was linear over three orders of magnitude with detection limits (S/N=3) ranging from 3 x 10(-8) to 2 x 10(-7)gml(-1) for the analytes. The method has been successfully applied to the analysis of real sample, with satisfactory results.     

Determination of active components in rosemary by capillary electrophoresis with electrochemical detection

Rosemary (Rosmarinus officinalis L.) is a spice and medicinal herb widely used around the world. Among natural antioxidants, rosemary has been widely accepted as one of the spices with the highest antioxidant activity. A capillary electrophoresis method for the determination of its active components using electrochemical detection was developed. Effects of several important factors were investigated to acquire the optimum conditions. The detection electrode was a 300 microm carbon disc electrode at a working potential of +0.90 V (versus SCE). The analytes can be well separated with 25 min in a 75 cm length fused-silica capillary at a separation voltage of 16 kV in an 80 mmol/l borate buffer (pH 9.0). The current response was linear over about three orders of magnitude with detection limits (S/N=3) ranging from 2 x 10(-7) to 1 x 10(-6)g/ml for all the analytes. The method was successfully used in the analysis of rosemary with relatively simple extraction procedures, and the assay results were satisfactory.     

灯盏花的化学成分研究

自灯盏花 [Erigeronbreviscapus(Vant.)Hand . Mazz .]全草乙醇提取物中分离得到 7个化合物 ,经理化性质和光谱分析分别鉴定为 :芹菜素 (1) ;山奈酚 (2 ) ;对羟基苯甲酸 (3) ;3 ,4 二羟基苯甲酸 (4 ) ;槲皮素 (5 ) ;木犀草素 (6 ) ;4′ 羟基黄芩素 (7)。其中山奈酚和槲皮素是首次从该植物中分离得到。     

Determination of four polyphenolic active ingredients from a pharmaceutical preparation by capillary zone electrophoresis with amperometric detection

The simultaneous determination of four active ingredients in Nao Xue Shuan Tablets, including ferulic acid, protocatechuic aldehyde, caffeic acid and protocatechuic acid was performed by capillary zone electrophoresis with amperometric detection (CZE-AD). The effects of working electrode potential, pH and concentration of running buffer, separation voltage and injection time on CZE-AD were investigated. Under the optimum conditions, the four analytes could be perfectly separated within 18min. A 300mum diameter carbon-disc electrode had a good response at +0.95V (vs. SCE) for the four analytes. The response was linear over three orders of magnitude with detection limits (S/N=3) as low as 10(-8) or 10(-9)g/mL for the analytes. The assay results were satisfactory with recoveries in the range of 85.2-93.0% and RSDs less than 3.3%.     

高效液相色谱法测定灯盏细辛提取物中两种有效成分的含量

目的：探讨灯盏细辛提取物中野黄芩苷、3,4-二-O-咖啡酰奎宁酸2种有效成分含量的高效液相色谱测定方法。方法：色谱柱选择Dikma Kromasil C18（250mm×4.6mm,5.0μm）,流动相选择乙腈：0.2%磷酸溶液=30∶70,检测波长为332nm,柱温为35℃。结果：野黄芩苷的线性回归方程为Y=1.554×103-0.201×102（r=0.9997）;3,4-二-O-咖啡酰奎宁酸的线性回归方程为Y=1.724×103-0.081×102（r=0.9996）;野黄芩苷在0.0341～1.3121μg范围内,3,4-二-O-咖啡酰奎宁酸在0.0592～2.3426μg范围内呈良好的线性关系。结论：采用高效液相色谱法同时测定灯盏细辛提取物中2种有效成分的含量,操作简单,精密度高,重现性好,可以作为灯盏细辛提取物中这2种有效成分的定量分析方法。     

Determination of flavonoids in Portulaca oleracea L. by capillary electrophoresis with electrochemical detection

A general method based on capillary electrophoresis with electrochemical detection (CE-ED) was developed for identification and determination of five flavonoids (kaempferol, apigenin, myricetin, quercetin and luteolin) in plant species. Running buffer, pH and concentration, separation voltage, injection time and detection potential were investigated to acquire the optimum conditions. The working electrode was a 500 microm diameter carbon disc electrode positioned opposite the outlet of capillary. At room temperature, the five flavonoids could be well separated within 21 min in a 60 cm length capillary at a separation voltage of 19.5 kV with 50 mM Na2B4O7-100 mM NaH2PO4 (pH 8.50) as the running buffer. The relationship between peak currents and analyte concentrations was linear over about two orders of magnitude, and the detection limits (S/N=3) were ranging from 0.12 to 0.21 microg/ml for all analytes. The optimized CE-ED method was employed to analyze the above flavonoids in different parts of Portulaca oleracea L.     

Simultaneous determination of neomycin sulfate and polymyxin B sulfate by capillary electrophoresis with indirect UV detection

A simple and rapid capillary electrophoresis method, with indirect UV detection, for the simultaneous determination of neomycin sulfate and polymyxin B sulfate in pharmaceutical formulations was developed. Critical parameters such as pH, buffer composition and concentration, voltage and injection time have been studied to evaluate, how they affect responses, such as resolution and migration times. Separation was performed on a fused silica capillary with 50 microm i.d. and 27 cm total length at an applied voltage of 6 kV with a 15 mM phosphate run buffer (pH 5.0) containing 40 mM N-(4-hydroxy-phenyl)acetamide and 50 mM tetradecylammonium bromide (TTAB). The detection wavelength was set at 280 nm. Quantitative analysis was validated by testing the reproducibility of the method, giving a relative standard deviation less than 0.4 and 2.4% for the repeatability of migration time and corrected peak area, respectively. Accuracy was tested by spiking eye-ear formulations with standards and the recoveries of neomycin sulfate and polymyxin B sulfate were found to be between 97.44-103.18% and 96.85-101.68%, respectively. Linearity of neomycin sulfate and polymyxin B sulfate were obtained in the ranges of 17-682 and 24-608 microg/mL, respectively, with r(2) values above 0.999. The established TLC-densitometric method was applied to evaluate the proposed CE method, and comparable results were obtained by using CE with much shorter analysis time and a small quantity of solvents consumed. The developed method is also the first report on the simultaneous determination of neomycin sulfate and polymyxin B sulfate in pharmaceutical preparations by CE.     

HPLC法同时测定益气复脉方中3种活性成分的含量

目的建立同时测定益气复脉方中芍药苷、甘草苷和甘草酸含量的方法。方法采用HPLC法测定芍药苷、甘草苷和甘草酸的含量,Purospher RP C_(18)色谱柱(250mm×4.6mm,5μm);乙腈-1mL·L~(-1)磷酸进行梯度洗脱;流速为1.0mL·min~(-1);检测波长为230和237nm;柱温为30℃;进样量为10μL。结果芍药苷、甘草苷和甘草酸的质量浓度分别在0.02~0.31,0.02~0.32和0.02~0.30mg·mL~(-1)范围内线性关系良好,r值均为0.999 9,3种成分的平均回收率分别为97.98%,98.23%和97.49%,RSD值均小于3.00%。结论该方法简便、准确、重复性好,可用于益气复脉方的内在质量控制。     

Simultaneous determination of five main active constituents of Erigeron multiradiatus by HPLC-DAD-MS

An HPLC-DAD-MS method was developed for simultaneous determination of the five major active constituents in Erigeron multiradiatus (Wall.) Benth, namely 6'-O-cafferylerigeroside (1), scutellarin (2), apigenin-7-O-beta-d-glucuronide (3), apigenin (4) and kaempferol (5), respectively. They were identified by ESI-MS and comparisons with literature. A comprehensive validation of the method included tests of sensitivity, linearity, precision and accuracy. The linear regressions were acquired with r>0.999. The precision was evaluated by intra- and inter-day assays, and relative standard deviation (R.S.D.) values were reported within 2.7%. The recovery studies for the quantified compounds were observed in the range of 95.3-102.4% with R.S.D. values less than 2.3%. The overall procedure may be suitable for the qualitative and quantitative analyses of a large number of E. multiradiatus samples. Hierarchical clustering analysis based on the characteristics of the 5 investigated compound peaks in HPLC profiles showed that 18 samples were divided into 2 main clusters. The clusters corresponded to their content. The five constituents in E. multiradiatus are generally regarded as an index for the quality assessment of this herb.     

高效液相色谱-电雾式检测器法同时测定川芎茶调颗粒中7个活性成分

目的:建立高效液相色谱法联合电雾式检测器同时测定川芎茶调颗粒中阿魏酸、甘草酸、阿魏酸松柏酯、洋川芎内酯A、Z-藁本内酯、欧前胡素及异欧前胡素的含量.方法:采用Agilent Eclipse XDB-C18(250 mm×4.6 mm,5 pm)色谱柱,以10 mmol·L-1甲酸铵(A)-80％乙腈(B)为流动相,梯度...     

Determination of glycosides and sugars in Moutan Cortex by capillary electrophoresis with electrochemical detection

A method based on capillary electrophoresis with electrochemical detection has been developed for the separation and determination of paeoniflorin, sucrose, paeonoside, glucose, and fructose in Moutan Cortex for the first time. Effects of several important factors such as the concentration of NaOH, separation voltage, injection time, and detection potential were investigated to acquire the optimum conditions. The detection electrode was a 300 microm diameter copper disc electrode at a working potential of +0.60 V (versus saturated calomel electrode (SCE)). The five analytes can be well-separated within 12 min in a 40 cm length fused silica capillary at a separation voltage of 12 kV in a 75 mM NaOH aqueous solution. The relation between peak current and analyte concentration was linear over about 3 orders of magnitude with detection limits (S/N = 3) ranging from 0.9 to 1.3 microM for all analytes. The proposed method has been successfully applied to monitor glycoside and sugar contents in the real plant samples with satisfactory assay results.     

毛细管区带电泳法同时测定冬虫夏草中多种核苷及其碱基成分含量

目的探讨毛细管电泳法研究天然冬虫夏草用不同溶剂提取的效果及测定其中核苷及其碱基含量的可行性。方法采用Unimicro毛细管电泳仪 ,石英毛细管柱 ( 6 0cm× 75 μmI D ) ,运行缓冲液分别为 36mmol/L硼砂 1 5mmol/L磷酸二氢钠 ( pH 8 9) ,分离温度 2 0℃ ,分离电压1 5kV ,测定波长 2 5 4nm ,虹吸进样 1 5s。结果基线分离冬虫夏草不同溶剂提取的样品。腺苷、尿苷、尿嘧啶和次黄嘌呤含量分别为 0 30 %、0 2 8%、0 1 4 %、0 0 5 % ;平均回收率分别为 96 4 %、98 2 %、94 3%、95 1 % ;RSD分别为 0 89%、1 2 %、1 4 %、1 8% (n =6 )。结论该法准确 ,快速 ,灵敏度高 ,对名贵药冬虫夏草的质量控制有较重要意义。     

Simultaneous determination of hydrochlorothiazide and several angiotensin-II-receptor antagonists by capillary electrophoresis

We have investigated the capability of the capillary zone electrophoretic (CZE) and micellar electrokinetic capillary chromatographic (MEKC) methods to simultaneously separate hydrochlorothiazide and six angiotensin-II-receptor antagonists (ARA-IIs): candesartan, eprosartan mesylate, irbesartan, losartan potassium, telmisartan, and valsartan. The CZE and MEKC methods are suitable for the qualitative and quantitative determination of combined HCT/ARA-IIs in pharmaceutical formulations. Depending on the ARA-II, at least one of the two methods can be used for each combination. The two methods have been validated in terms of their linearity of response, reproducibility, and accuracy.     

毛细管区带电泳分离／安培检测儿茶酚胺药物

目的:建立毛细管区带电泳(CZE)分离/安培检测左旋多巴(DP)、多巴胺(DA)、肾上腺素(EP)、去甲肾上腺素(NE)、5-羟色胺(5-HT)5种儿茶酚胺类神经递质(CAs)药物的方法。方法:采用石英毛细管柱(60 cm×50μm),含8 mmol·L~(-1)对-(季铵盐)杯[4]芳烃(QAC4A)的125 mmol·L~(-1)磷酸氢二钠溶液(Na_2HPO_4,pH 7.0)为缓冲液,分离电压6 kV,分离 CAs 并利用 Cu 微粒修饰碳纤维电极检测。结果:优化条件下5种 CAs 被基线分离,回收率为92%～105%,5种 CAs 在0.2～1000μmol·L~(-1)浓度内呈良好线性关系,最低检测限为0.2 μmol·L~(-1)。应用该方法成功地测定了5种 CAs 注射液、正常人和嗜铬细胞瘤患者尿样。结论:本法用于分离并检测 CAs 药物,具有准确、灵敏、简便的特点,适合该类药品分析。     

高效液相色谱-蒸发光散射检测法同时测定壮药愈疡散中3种主要活性成分的含量

目的：建立测定壮药愈疡散中黄芪甲苷、紫丁香苷与长梗冬青苷含量的方法。方法：采用高效液相色谱-蒸发光散射检测（HPLC-ELSD）法,色谱柱为Shim-pack CLC-ODS（150 mm×4.6 mm,5 μm）,以乙腈-水为流动相,梯度洗脱,流速1 mL·min^-1,柱温20℃;检测参数为漂移管温度为90℃,雾化器温度为45℃,气体流量为1.60 L·min^-1,增益值为1.0。结果：黄芪甲苷、紫丁香苷和长梗冬青苷分别在0.46~2.76 μg（r=0.999 2）、0.70~4.20 μg（r=0.996 2）和1.06~6.36 μg（r=0.998 6）范围内,其峰面积的自然对数与进样量的自然对数呈良好的线性关系;平均加样回收率分别为95.89%、95.49%和97.66%,RSD为2.79%、3.96%和3.60%（n=6）。结论：该法简便快速、重复性好,结果准确,可作为壮药愈疡散的质量控制方法。     

~(45)Ca同位素技术用于灯盏细辛中抗钙活性有效组分的研究

灯盏细辛(Erigeron breviscapus(Vant.)Hand-Mazz),主要含以野黄芩苷为主的黄酮、吡喃酮、酚酸类等化合物.具有散寒解表,活血舒筋,止痛消积等功效,用于治疗血栓性及心脑血管疾病等[1].越来越多研究报道灯盏细辛具青光眼视神经保护作用[2,3].最近研究表明,钙通道阻滞剂能与细胞膜钙通道结合,减少钙离子内流,扩张血管,从而改善视盘血供,阻断视网膜神经节细胞凋亡,起到视神经保护作用,在青光眼的治疗方面具广阔的应用前景[4,5].另据报道灯盏花素能抑制NA引起的兔胸主动脉内Ca2 浓度升高[6].基于此,本研究采用45Ca同位素示踪技术[7],观察了灯盏细辛中4种组分对大鼠胸主动脉电压依赖性钙通道(VDC)Ca2 内流的影响,以探讨其钙拮抗活性.     

Determination of honokiol and magnolol in cortex Magnoliae Officinalis by capillary electrophoresis with electrochemical detection

Capillary electrophoresis with electrochemical detection has been employed for the determination of honokiol and magnolol in Cortex Magnoliae Officinalis (i.e. Magnolia Bark) for the first time. Effects of several important factors such as the concentration and the acidity of the running buffer, separation voltage, injection time, and detection potential were investigated to acquire the optimum conditions. The detection electrode was a 300 microm diameter carbon disc electrode at a working potential of +0.90 V (versus saturated calomel electrode (SCE)). The two analytes can be well separated within 6 min in a 40 cm length fused silica capillary at a separation voltage of 18 kV in a 50mM borate buffer (pH 9.2). The relation between peak current and analyte concentration was linear over about three orders of magnitude with the detection limits (S/N=3) of 0.38 and 0.51 microM for honokiol and magnolol, respectively. The proposed method has been successfully applied to monitor the two bioactive constituents in the real plant samples with satisfactory assay results.     

毛细管区带电泳法测定司巴沙星和茶碱的血药浓度

目的：建立用毛细管区带电泳法同时测定司巴沙星和茶碱血药浓度方法。方法血 离心后，分离血清，直接进样测定，用２０ｍｍｏｌ／Ｌ硼砂缓冲液分离，２９８ｎｍ为测定波长，外标峰面积法定量。结果：血清中司巴沙星和茶碱分离良好，分别在１．２～１２．５μｇ／ｍｌ、３．８～３７．６μｇ／ｍｌ的浓度范围内线性关系良好。其血清最低检测浓度分别为０．８和２．５μｇ／ｍｌ。结论本法可作为同巴沙得和茶碱药物动力学研究时的血药     

毛细管区带电泳电化学检测人体尿样中的双氯灭痛、氯丙嗪

目的建立毛细管区带电泳法测定人体尿样中双氯灭痛、氯丙嗪含量的方法。方法采用弹性石英毛细管（31.5cm,25μmi.d.,360μmo.d.）,以4.90×10-3mol/LNa2HPO4-7.80×10-3mol/LNaH2PO4（pH=7）为缓冲液,10kV分离电压,5kV电渗进样10s,碳纤维电极工作电极（长200μm,直径8μm）,检测电势0.72V。结果双氯灭痛、氯丙嗪两种药物分别在9.90×10-6～5.00×10-4mol/L（r=0.9982）、5.0×10-7～1.0×10-4mol/L（r=0.9998）范围内表现出良好的线性,其回收率分别为104%,96.0%。结论该方法简便、快速、准确,可作为人体尿样中双氯灭痛、氯丙嗪的分离检测方法。     

Differentiation of Swertia Mussotii Franch from Artemisiae Capillaris Herba by capillary electrophoresis with electrochemical detection

A high-performance capillary electrophoresis (CE) with electrochemical detection (ED) method is developed for differentiation of Swertia Mussotii Franch from Artemisiae Capillaris Herba in this work. Swertia Mussotii Franch contains a great deal of swertiamarin and mangiferin that are not present in Artemisiae Capillaris Herba, whereas Artemisiae Capillaris Herba consists of abundant chlorogentic acid. Therefore, determining their swertiamarin, mangiferin and chlorogentic acid contents can differentiate these two crude herbs. Operated in a wall-jet configuration, a 300 microm diameter carbon-disk electrode was used as the working electrode, which exhibits good response at +1000 mV (versus SCE) for the three analytes. With a separation voltage of 14 kV, the three analytes were separated within 14 min in a 52 cm length capillary in 50 mmol/l borax buffer (pH 9.2). The system was demonstrated good stability and reproducibility with an R.S.D. of less than 5% for both migration time and peak current. This method was successfully used to analyze and identify the crude herbs with satisfactory assay results.