反相高效液相色谱法测定不同产地紫苏子和白苏子中乌索酸和齐墩果酸

建立反相高效液相色谱（RP—HPLC）法测定了紫苏子和白苏子中乌索酸和齐墩果酸的含量。色谱柱为Kromasil C18柱（250mm×4．6mm,5um）,光电二极管阵列检测（PAD）,流动相甲醇一水体积比为87：13,检测波长210nm,流速0．8mL／min,柱温28℃。乌索酸在10～400ug／mL,齐墩果酸在5～200ug／mL内呈现良好的线性关系,平均加样回收率分别为100．6％和98．7％,相对标准偏差（RSD）分别为1．6％和1．2％。     

HPLC法同时测定鸡矢藤中熊果酸和齐墩果酸的含量

目的：建立高效液相色谱法同时测定鸡矢藤中熊果酸和齐墩果酸的含量。方法：色谱柱为DiamonsilTM C18（250×4.6 mm,5滋m）,流动相为甲醇-0.05mol.L-1磷酸溶液（93∶7）为流动相,检测波长210 nm,进样量10滋L。结果：熊果酸在21.28滋g.mL-1～212.8滋g.mL-1范围内线性关系良好（r=0.9997）,平均回收率为97.79%（RSD=1.30%）;齐墩果酸在24.02滋g.mL-1～240.2滋g.mL-1范围内线性关系良好（r=0.9996）,平均回收率为98.36%（RSD=1.11%）。结论：该方法简便、灵敏、准确,可用于鸡矢藤药材的质量评价。     

小花清风藤的化学成分研究

利用Sephadex LH-20,硅胶柱色谱和硅胶制备薄层色谱等分离方法反复分离纯化,从小花清风藤（Sabia parviflora Wall．ex Roxb）中分离得到9个化合物,通过波谱数据分析分别鉴定它们为二十五烷酸（1）、木栓酮（2）、5-氧阿朴菲碱（3）、3-氧化齐墩果酸甲酯（4）、齐墩果酸（5）、羽扇豆-20（29）-烯-3-酮（6）、羽扇豆醇（7）、β-谷甾醇（8）、脱镁叶绿甲酯酸（9）。化合物2,6、7、9首次从该植物中分离得到。     

反相高效液相色谱法测定鹿衔草中熊果酸和齐墩果酸的含量

建立反相高效液相色谱法测定鹿衔草中熊果酸和齐墩果酸含量,并进行方法学考察.采用Kromasil C18分析柱(4.6 mm×250 mm,5 μm);流动相为甲醇-水(88:12);柱温25 ℃;流速0.8 mL/min;检测波长210 nm.熊果酸进样量在0.922～18.44 μg,齐墩果酸进样量在0.506～10.12 μg范围内呈良好线性关系,其中熊果酸平均回收率为98.5%,RSD为2.13%(n=6);齐墩果酸平均回收率为 101.3%,RSD为1.69%(n=6).本方法使鹿衔草中主要成分熊果酸和齐墩果酸达到基线分离,操作简便、结果可靠,可为鹿衔草质量控制和评价提供有效手段.     

威灵仙药材中三萜苷元齐墩果酸含量的HPLC测定

为探索可用于威灵仙药材质量控制的三萜苷元齐墩果酸HPLC含量测定方法,用HPLC法测定了威灵仙药材中齐墩果酸含量。色谱柱:Kromasil C18(4.6×150 mm,5μm),检测波长:220 nm,流动相:甲醇-水(95:5),流速:0.5 mL/min。齐墩果酸在0.004~0.015 mg范围内线性关系良好,回归方程为:Y=437954101.73X 144588.26(R=0.9991),平均回收率为:98.33%,RSD=1.76%。样品中齐墩果酸含量分别为:东北铁线莲0.2281%;威灵仙0.1260%、棉团铁线莲0.1851%。本方法快速、准确、灵敏度高、重现性好,适用于对威灵仙中齐墩果酸的含量进行测定。     

红背桂花化学成分研究

为了对红背桂花(Excoecaria cochinchinensis Lour.)的化学成分进行研究,采用了硅胶柱色谱、Sephadex LH-20凝胶柱色谱、反相ODS柱色谱等色谱方法,对红背桂花的80%乙醇提取物进行系统的分离,得到了10个单体化合物,通过理化性质与波谱数据分析,分别鉴定为:齐墩果酸(oleanolic acid,1)、2α,3β,23-三羟基齐墩果-12-烯-28-酸(arjunolic acid,2)、山奈酚(kaempferol,3)、槲皮素(quercetin,4)、高山黄芩(isoscutellarein,5)、6-甲氧基-7-羟基香豆素(scopoletin,6)、3,4,5-三羟基苯甲酸(3,4,5-trihydroxybenzoic acid,7)、间苯三酚(phloroglucinol,8)、β-谷甾醇(β-sitosterol,9)和胡萝卜苷(daucosterol,10)。化合物2,5和8为首次从海漆属植物中分离得到,化合物1,4和6为首次从红背桂花植物中分离得到。     

小花草玉梅化学成分研究

目的：研究银莲花属植物小花草玉梅的化学成分。方法：采用硅胶柱色谱,凝胶柱色谱,反相柱色谱并结合制备高效液相色谱等技术分离纯化单体化合物,并根据理化性质及光谱数据鉴定结构。结果：分离并鉴定了4个化合物,分别是常春藤皂苷元-28-O-β-D-吡喃葡萄糖酯苷（1）、3-O-β-D-吡喃葡萄糖-（1→2）-α-L-吡喃阿拉伯糖-齐墩果酸皂苷元-28-O-α-L-吡喃鼠李糖-（1→4）-β-D-吡喃葡萄糖-（1→6）-β-D-吡喃葡萄糖酯苷（2）、3-O-β-D-吡喃葡萄糖-（1→2）-α-L-吡喃阿拉伯糖-常春藤皂苷元-28-O-α-L-吡喃鼠李糖-（1→4）-β-D-吡喃葡萄糖-（1→6）-β-D-吡喃葡萄糖酯苷（3）和3-O-β-D-吡喃核糖-（1→3）-α-L-吡喃鼠李糖-（1→2）-α-L-吡喃阿拉伯糖-常春藤皂苷元-28-O-α-L-吡喃鼠李糖-（1→4）-β-D-吡喃葡萄糖-（1→6）-β-D-吡喃葡萄糖酯苷（4）。结论：化合物1为首次从银莲花属植物中分离得到,2-4为首次从小花草玉梅中分离得到。     

龙胆草地上部分的化学成分

为研究龙胆草（Gentiana scabra Bunge）地上部分的化学成分,利用硅胶柱色谱对其乙醇提取物进行分离纯化,根据理化性质和波谱方法鉴定了化合物的结构。分离得到β-香树脂醇（1）、β-谷甾醇（2）、β-香树脂醇乙酸酯（3）、乌苏醇（4）、齐墩果酸（5）、6-去甲氧基-7-甲基茵陈色原酮（6）。其中化合物6为首次从龙胆科植物中分离得到的具有2-苯氧基色原酮骨架的天然酚类成分,本文对其1D和2DNMR谱的特征及EI质谱主要碎片的可能裂解方式进行了讨论,利用单晶X-射线衍射方法对其晶体结构进行了分析,为该类天然产物的结构表征提供了依据。     

齐墩果酸对APP/PS-1双转基因AD小鼠的神经保护作用研究

摘要 目的：研究齐墩果酸（Oleanolic Acid，OA）对APP/PS-1双转基因阿尔茨海默病（Alzheimer''s disease，AD）小鼠模型神经保护作用及机制。方法：选取6月龄APP/PS-1雄性小鼠21只，随机分为模型组（0.5% CMC-Na）、阳性组（多奈哌齐组，0.7 mg?kg^-1）、齐墩果酸组（10 mg?kg^-1）每组7只，6月龄同背景SPF级C57BL/6小鼠7只为对照组。灌胃8周之后通过Morris水迷宫实验观察小鼠学习记忆能力的改变，HE染色观察神经元细胞形态，ELISA检测血清中Aβ1-42含量；免疫组化检测Aβ_1-42、APP、Iba1蛋白表达情况；Western blot检测APP、Iba1蛋白表达水平。结果：（1）对照组，模型组，阳性组及齐墩果酸组进入有效区域次数分别为7.00±2.09，1.00±0.89，3.67±1.97，4.33±2.50，与模型组相比，对照组，阳性组，齐墩果酸组均有统计学意义（P<0.05）；（2）血清Aβ_1-42含量按上述顺序依次为4.98±0.25，2.50±0.66，4.63±0.73，4.36±0.97，与模型组相比，对照组，阳性组，齐墩果酸组均有统计学意义（P<0.05）；（3）免疫组化结果显示与模型组相比，对照组，阳性组，齐墩果酸组Aβ_1-42、APP、Iba1蛋白阳性细胞数减少；（4）WB结果：对照组，模型组，阳性组，齐墩果酸组APP蛋白相对表达量分别为0.52±0.17，1.38±0.35，0.89±0.25，0.93±0.27；这四组的IBA1蛋白相对表达量分别为0.98±0.34，1.79±0.74，1.06±0.61，0.88±0.49，与模型组相比，野生对照组，阳性组，齐墩果酸组APP、IBA1蛋白相对含量有统计学意义（P<0.05）。结论：齐墩果酸组可以改善APP/PS-1模型小鼠记忆力及认知功能，降低海马神经元的损伤，并通过下调Aβ_1-42、APP、Iba1蛋白的表达水平来发挥保护神经作用。     

南丹参三萜类化学成分的研究

为研究南丹参根中三萜类化学成分,该文采用95%乙醇浸泡提取,依次应用D101大孔吸附树脂、MCI、硅胶柱色谱、ODS及反相高效液相色谱方法进行分离纯化,根据理化性质及波谱数据鉴定其化合物的结构。结果表明:从南丹参根提取物中分离得到9个化合物,分别鉴定为齐墩果酸(1)、2α,3α-二羟基-12-烯-28-齐墩果酸(2)、乌苏酸(3)、2α,3α-二羟基-12-烯-28-乌苏酸(4)、2α,3β-二羟基-12-烯-28-乌苏酸(5)、2α,3β,19α-三羟基-12-烯-28-乌苏酸(6)、2α,3β,24-三羟基-12-烯-28-乌苏酸(7)、psiguanin A(8)、正十六烷酸(9)。化合物1-9为首次从该植物中分离得到。     

Reversed-phase liquid chromatography analysis of imatinib mesylate and impurity product in Glivec capsules

The reversed-phase high-performance liquid chromatographic (RP-HPLC) method has been developed and validated for the simultaneous determination of imatinib mesylate and of the impurity product in Glivec capsules (Novartis, Switzerland). Separations were performed on a X Terra 150 mm x 4.6 mm, 5 microm particle size column at 25 degrees C. The mobile phase was a mixture of methanol-water-triethylamine (25:74:1, v/v/v) with flow rate of 1.0 ml min(-1). pH value of water-triethylamine (TEA) was adjusted to 2.4 with orthophosphoric acid before adding of methanol. UV detection was performed at 267 nm. Acetaminophen was used as an internal standard. The method was validated statistically for its selectivity, linearity, precision, accuracy and robustness. Due to its speed and accuracy, the method may be used for quality control analyses.     

桂南地区苦玄参药材RP-HPLC指纹图谱研究

采用反相HPLC法测定了广西梧州、龙州、苍梧、越南等多产地12批苦玄参药材指纹图谱,并对不同产地的苦玄参指纹图谱进行比较。色谱条件为:LunaC18(4.6×250mm,5μm)色谱柱,乙腈-水梯度洗脱,流速1.0mL/min,检测波长254nm,柱温25℃。结果12批苦玄参样品指纹图谱共标定了16个分离度良好的共有峰,方法的精密度、稳定性、重复性均符合国家相关规定,可作为控制苦玄参药材质量的定性标准。     

HPLC法测定藜豆中左旋多巴的含量

采用HPLC法建立测定藜豆中左旋多巴含量的方法。色谱柱为岛津C-18甲基硅烷柱(250 min×4.6mm,5μm),流动相为甲醇:0.1 mol／L醋酸溶液(25:75,v／v),以硫唑嘌呤为内标物,检测波长280 nm,流速1.0 mL／min,柱温为室温,供试品的前处理方法为微波辅助提取法。结果表明,该方法准确度高,线性关系好,重现性好,平均加样回收率为97.34％,RSD 1.01％(n=5),结果满意。     

Quantitation of itopride in human serum by high-performance liquid chromatography with fluorescence detection and its application to a bioequivalence study

A new method was developed for determination of itopride in human serum by reversed phase high-performance liquid chromatography (HPLC) with fluorescence detection (excitation at 291 nm and emission at 342 nm). The method employed one-step extraction of itopride from serum matrix with a mixture of tert-butyl methyl ether and dichloromethane (70:30, v/v) using etoricoxib as an internal standard. Chromatographic separation was obtained within 12.0 min using a reverse phase YMC-Pack AM ODS column (250 mm x 4.6 mm, 5 microm) and an isocratic mobile phase constituting of a mixture of 0.05% tri-fluoro acetic acid in water and acetonitrile (75:25, v/v) flowing at a flow rate of 1.0 ml/min. The method was linear in the range of 14.0 ng/ml to 1000.0 ng/ml. The lower limit of quantitation (LLOQ) was 14.0 ng/ml. Average recovery of itopride and the internal standard from the biological matrix was more than 66.04 and 64.57%, respectively. The inter-day accuracy of the drug containing serum samples was more than 97.81% with a precision of 2.31-3.68%. The intra-day accuracy was 96.91% or more with a precision of 5.17-9.50%. Serum samples containing itopride were stable for 180.0 days at -70+/-5 degrees C and for 24.0 h at ambient temperature (25+/-5 degrees C). The method was successfully applied to the bioequivalence study of itopride in healthy, male human subjects.     

Enantiomeric separation of norgestrel by reversed phase high-performance liquid chromatography using eluents containing hydroxypropyl-beta-cyclodextrin in stereoselective skin permeation study

The chiral separation of norgestrel enantiomers using reversed-phase high-performance liquid chromatography (RP-HPLC) was studied with hydroxypropyl-beta-cyclodextrin (HP-beta-CD) as chiral mobile phase additive. The effect of mobile phase composition, concentration of HP-beta-CD and column temperature on enantioselective separation were investigated. The quantification properties of the developed RP-HPLC method were examined. A baseline separation of norgestrel enantiomers was achieved on a Agilent ZORBAX Eclipse XDB-C8 column (150 mm x 4.6 mm i.d., 5 microm). The mobile phase was a mixture of acetonitrile and phosphate buffer (pH 5.0, 20 mM) containing 25 mM HP-beta-CD (30:70, v/v) with a flow rate of 1.0 ml/min. The UV detector was set at 240 nm. Calibration curves were linear (n=8) in the range of 0.2-25 microg/ml, the limit of detection and quantitation were 0.10 and 0.20 microg/ml, respectively, for racemic norgestrel. The values of RSD of repeatability and intermediate precision for spiked sample were less than 4.8%. The method was successfully applied to the enantioselective determination of this drug in stereoselective skin permeation study.     

Determination of spinosin in rat plasma by reversed-phase high-performance chromatography after oral administration of Suanzaoren decoction

A sensitive, simple, and accurate method for determination of spinosin in rat plasma with sulfamethoxazole (SMZ) as internal standard was developed using RP-HPLC with UV detection. Sample preparations were carried out by protein precipitation with acetonitrile, followed by the evaporation of the acetonitrile to dryness. The resultant residue was then reconstituted in mobile phase and injected onto a Hypersil C(18) (200 x 4.6 mm I.D., 5 microm) analytical column. The mobile phase consisted of acetonitrile-water (15:85, v/v) with 1% glacial acetic acid. The assay was shown to be linear over the range of 18.07-903.5 ng/ml (R(2)=0.995). Mean recovery was determined as 93.6%. Within- and between-day precisions were 相似文献   

Ion-pair reverse-phase high performance liquid chromatography method for determination of Huperzine-A in beagle dog serum

Huperzine-A (Hup-A), a biologically potent, reversible acetylcholinesterase inhibitor for the treatment of Alzheimer disease (AD) in China, has very low blood concentration. In order to study the pharmacokinetics of newly developed Hup-A transdermal patches in animal, a rapid and sensitive ion-pair reverse-phase high performance liquid chromatography (RP-HPLC) method for the determination of Hup-A in beagle dog serum using mebendazole as internal standard has been developed and validated. The analyte and internal standard were extracted from serum using chloroform-isopropanol (95:5, v/v), analyzed on a C (18) column (5 microm, 150 mm x 4.6 mm i.d.) with a mobile phase consisting of methanol-water-glacial acetic acid (50:48.5:1.5, v/v/v), using sodium dodecylsulfonate as an ion-pair reagent, and detected with UV detector at 313 nm. The chromatographic run time was within 15 min. The assay was linear over the concentration range of 1-12 ng/ml and intra- and inter-day precision over this range was not more than 12.8%. The limit of quantification in serum was 1 ng/ml. The method was successfully applied to characterize the Hup-A concentration-time profiles and study the single and multiple doses phamacokinetics of Hup-A transdermal patches in beagle dogs. The pharmacokinetic study results showed that Hup-A patches has the characteristic of sustained or controlled drug release in vivo.     

Determination of aesculin in rat plasma by high performance liquid chromatography method and its application to pharmacokinetics studies

A sensitive and reproducible high performance liquid chromatography method with UV detection was described for the determination of aesculin in rat plasma. After deproteinization by methanol using metronidazole as internal standard (I.S.), solutes were evaporated to dryness at 40 degrees C under a gentle stream of nitrogen. The residue was reconstituted in 100 microl of mobile phase and a volume of 20 microl was injected into the HPLC for analysis. Solutes were separated on a Diamonsil C18 column (250 mm x 4.6 mm i.d., 5 microm particle size, Dikma) protected by a ODS guard column (10 mm x 4.0 mm i.d., 5 microm particle size), using acetonitrile-0.1% triethylamine solution (adjusted to pH 3.0 using phosphoric acid) (10:90, v/v) as mobile phase (flow-rate 1.0 ml/min), and wavelength of the UV detector was set at 338 nm. No interference from any endogenous substances was observed during the elution of aesculin and internal standard (I.S., metronidazole). The retention times for I.S and aesculin were 10.4 and 12.4 min, respectively. The limit of quantification was evaluated to be 57.4 ng/ml and the limit of detection was 24.0 ng/ml. The method was used in the study of pharmacokinetics of aesculin after intraperitoneal injection (i.p.) administration in rats.     

低分子量羊胎提取液RP-HPLC分析方法的初步研究

采用RP -HPLC法在ODSHypersil柱上分析低分子量羊胎提取物中的活性成分。通过多种色谱参数的比较 ,探讨了流动相、梯度、流速、温度、波长对混合未知成分分离的影响 ,获得最佳色谱条件。对其重现性进行考察 ,发现各峰保留时间的相对标准偏差均可控制在 0 .3%以下 ,峰面积则在 2 .0 %以内。说明该研究能够有效的针对动物提取的混合成分 ,建立一种准确可靠的RP -HPLC分析方法 ,为质量标准的控制提供了新的测定指标。