HPLC法同时测定利胆解毒胶囊中异土木香内酯和土木香内酯的含量

目的:建立高效液相色谱法(HPLC)同时测定利胆解毒胶囊中异土木香内酯和土木香内酯含量的方法.方法:采用Diamonsil Plus C18色谱柱(4.6 mm×250 mm,5μm),流动相为乙腈-0.05％磷酸溶液(50:50),流速1.0 mL·min-1,检测波长210 nm,柱温30.0℃.结果:异土木香内酯...     

果旭阿汤中土木香内酯和异土木香内酯的含量测定

［摘要］目的建立同时测定藏药果旭阿汤中土木香内酯和异土木香内酯含量的方法。方法通过优化提取方法,用气相色谱（GC）法同时测定藏药果旭阿汤中藏木香有效成分土木香内酯和异土木香内酯的含量。结果土木香内酯和异土木香内酯均在0.1～1.0 mg•mL-1 (n=5）范围内呈良好线性关系。土木香内酯回收率为97.25% (n=6),RSD=0.93%;异土木香内酯回收率为97.42% (n=6), RSD=0.90%。结论该方法可用于藏药果旭阿汤中土木香内酯和异土木香内酯的含量同时快速、准确测定。     

GC法测定额尔敦尼七味汤中异土木香内酯的含量

目的:建立额尔敦尼七味汤中异土木香内酯含量的测定方法。方法:选用气相色谱法。结果:异土木香内酯进样量在20.72ng～414.4ng范围内可靠,回归方程为Y=2154.6X+2199.6,r=0.9999。平均回收率(n=9)为99.25%,RSD为1.51%。结论:本法可用于额尔敦尼七味汤中异土木香内酯的含量测定,为一种结果准确、灵敏、可行的方法。     

总状土木香氯仿部位的化学成分研究

目的:研究总状土木香氯仿部位的化学成分。方法:采用硅胶、反相硅胶、SephadexLH-20凝胶柱色谱对样品进行分离纯化,根据理化性质和光谱分析鉴定化合物结构。结果:从总状土木香氯仿部位中分离得到6个化合物,经鉴定分别为11α,13-二羟基去氢闭鞘姜内酯(1)、11,13-二氢-7,11-去氢-13-羟基-3-脱氧愈创内酯C(2)、1β-羟基柯拉汀(3)、胡萝卜苷(4)、β-谷甾醇(5)、丁香树脂醇(6)。其中化合物1、2、3和6为首次从该属植物中分离得到。结论:本试验结果可为总状土木香的进一步研究提供依据。     

用HPLC法同时测定六味安消胶囊中土木香内酯、异土木香内酯、大黄素和大黄酚的含量

目的：建立HPLC法同时测定六昧安消胶囊中土木香内酯、异土木香内酯、大黄素和大黄酚含量的方法。方法：采用HPLC-二极管阵列检测器法（HPLC—DAD），色谱柱：Agilent Eclipse PlusC18柱（250mm×4.6mm，5μm），流动相：乙腈-0．04％甲酸（60：40），流速：1．0mL／min，检测波长：220nm，柱温：25℃，进样量：10μL。结果：土木香内酯、异土木香内酯、大黄素和大黄酚的线性范围分别是：2．46～98．4、2．48～99．2、2．56～102．4和2．58～103．2μg／mL，色谱响应线性关系良好。日内和日间RSD均〈2％；最低检测限分别为1．75、1．31、0．50和0．49μg／mL。稳定性结果为峰面积的RSD分别为1．29％、1．40％、1．30％和0．45％，在48h内稳定。加样回收率结果分别为99．0％、100．6％、98．7％和99．7％（n＝6），RSD分别为1．03％、0．62％、1．36％和0．26％。结论：本法快速、简便，重现性好，精密度高，测定结果准确可靠，可作为该制剂的定量测定方法。     

用HPLC法同时测定六昧安消胶囊中土木香内酯、异土木香内酯、大黄素和大黄酚的含量

目的:建立HPLC法同时测定六味安消胶囊中土木香内酯、异土木香内酯、大黄素和大黄酚含量的方法.方法:采用HPLC二极管阵列检测器法(HPLC-DAD),色谱柱:Agilent Eclipse Plus C18柱(250 mmX4.6mm,5μm),流动相:乙腈0.04%甲酸(60:40),流速:1.0 mL/min,检测波长:220nm,柱温:25℃,进样量:10μL.结果:土木香内酯、异土木香内酯、大黄素和大黄酚的线性范围分别是:2.46～98.4、2.48～99.2、2.56～102.4和2.58～103.2μg/mL,色谱响应线性关系良好.日内和日间RSD均＜2%;最低检测限分别为1.75、1.31、0.50和0.49μg/mL.稳定性结果为峰面积的RSD分别为1.29%、1.40%、1.30%和0.45%,在48h内稳定.加样回收率结果分别为99.0%、100.6%,98.7%和99.7%(n=6),RSD分别为1.03%、0.62%、1.36%和0.26%.结论:本法快速、简便,重现性好,精密度高,测定结果准确可靠,可作为该制剂的定量测定方法.     

HPLC法同时测定康妇软膏中异土木香内酯、土木香内酯、花椒毒酚、氧化前胡素、欧前胡素和蛇床子素

摘 要 目的：建立HPLC梯度洗脱法同时测定康妇软膏中异土木香内酯、土木香内酯、花椒毒酚、氧化前胡素、欧前胡素和蛇床子素的含量。 方法: 以Agilent TC C18（250 mm×4.6 mm,5 μm）为色谱柱,甲醇 0.1%甲酸溶液为流动相,梯度洗脱,流速0.9 ml·min^-1 ,检测波长分别为220 nm和300 nm,柱温35 ℃。 结果:异土木香内酯、土木香内酯、花椒毒酚、氧化前胡素、欧前胡素和蛇床子素分别在6.16～123.20 μg·m^-1 、3.78～75.60 μg·m^-1 、1.87～37.40 μg·m^-1 、4.06～81.20 μg·m^-1 、9.27～185.40 μg·m^-1 、13.89～277.80 μg·m^-1 范围内线性关系良好,相关系数分别为0.999 4,0.999 8,0.999 6,0.999 5,0.999 9和0.999 7;平均加样回收率分别为98.04%（RSD=1.06%）,97.10%（RSD=1.53%）,96.73%（RSD=0.90%）,98.92%（RSD=1.36%）,99.12%（RSD=0.83%）和100.27%（RSD=0.58%）。结论:该方法简便、准确,可用于康妇软膏质量标准的提高。     

HPLC-ELSD法测定杏香兔耳风中3个倍半萜内酯的含量

目的:建立HPLC-ELSD法同时测定药材杏香兔耳风中3个倍半萜内酯含量的方法.方法:采用Shim-pack CLC-ODS C18色谱柱 (150 mm×6.0 mm, 5 μm ) 以甲醇-2 %醋酸为流动相,梯度洗脱,蒸发光散射检测器 (ELSD) 检测.结果:被测定峰能够与其他组分达到基线分离,成分为8α-hydroxy-11αH-11,13-dihydro-zaluzanin C-3-O-β-D-glucopyranoside (1)、8α-hydroxy-11αH-11,13-dihydro-zaluzanin C (2) 和 zaluzanin C-3-O-β-D-glucopyranoside (3) 的线性范围分别为0.78～7.80 μg、0.46～4.64 μg和0.44～4.36 μg,平均回收率分别为103.5%、104.7%和98.9%, RSD分别为0.28%、0.13%和0.13%.结论: 该方法简便、准确,分离效果好,可用于对药材杏香兔耳风进行质量评价.     

木香药材的质量评价研究——HPLC法测定木香中2种倍半萜内酯的含量

目的 :建立木香药材的质量评价标准。方法 :采用高效液相色谱法测定了 2 2批药材及饮片中木香烃内酯和去氢木香内酯的含量。色谱柱为C18柱 (6 0mm× 15 0mm) ,流动相为甲醇 -水 (6 5∶35 ) ,检测波长为 2 2 5nm。结果 :定量方法简单 ,快速 ,准确。木香烃内酯和去氢木香内酯的回收率分别为 99 6 % ,98 4% ;RSD(n =5 )分别为 1 2 % ,1 8%。结论 :该方法可做控制木香质量用。     

HPLC法同时测定寒水石二十一味散中6种成分的含量

摘要：目的:建立HPLC法同时测定寒水石二十一味散中6种成分的含量。方法:采用Phenomenex Gemini C18色谱柱（250mm×4.6mm,5μm）,以乙腈为流动相A,0.1%磷酸为流动相B,梯度洗脱,流速:1.0 ml·min-1,检测波长:225 nm,进样量:10μl,柱温:30℃。结果:木香烃内酯、去氢木香内酯、土木香内酯、异土木香内酯、没食子酸和鞣花酸分别在4.60～184.00μg·ml-1（r=0.999 9）,5.72～228.60μg·ml-1（r=0.999 3）,5.69～227.60μg·ml-1（r=0.999 7）,5.52～220.80μg·ml-1（r=0.999 9）,4.75～190.00μg·ml-1（r=0.999 6）,4.70～187.80μg·ml-1（r=0.999 6）范围内线性良好,平均回收率分别为99.98%（RSD=0.50%）,99.41%（RSD=0.76%）,100.26%（RSD=0.98%）,99.66%（RSD=1.04%）,99.74%（RSD=0.98%）,99.96%（RSD=0.81%）（n=9）。结论:该方法结果准确、操作简便,重复性好,可用于寒水石二十一味散的质量控制。     

Five new sesquiterpene lactones from Inula hupehensis

Four new pseudoguaianolides (1–4), one new guaianolide (5), together with ten known compounds (6–15) were isolated from the aerial parts of Inula hupehensis. Their structures were elucidated mainly on the basis of 1D and 2D spectroscopic methods and circular dichroism analysis. In addition, compounds 1–10 and 13 were tested for their inhibitory effects against LPS-induced NO production in RAW264.7 macrophages. Compounds 2, 6, 8 and 9 exhibited significant inhibitory activities with IC₅₀ values in the range of 0.6–6.6 μM.     

Antiproliferative sesquiterpene lactones from the roots of Inula helenium

The MeOH extract of the roots of Inula helenium showed a high inhibitory activity for cell growth against MK-1, HeLa and B16F10 cell lines. Significant activity was found in the hexane-soluble fraction. From the hexane-soluble fraction, seven sesquiterpenes, namely, one germacrane (4beta,5alpha-epoxy-1(10),11(13)-germacradiene-8,12-olide), one elemane (igalane), and five eudesmanes (alantolactone, isoalantolactone, 11alpha,13-dihydroalantolactone, 11alpha,13-dihydro-isoalantolactone, 5-epoxyalantolactone) were isolated. In vitro antiproliferative activities of the isolates against MK-1, HeLa and B16F10 cells are reported.     

Sesquiterpenoids from Inula racemosa

Phytochemical research on the roots of Inula racemosa yielded nine sesquiterpenoids including a new nor-eudesmane-type sesquiterpenoid, 11,12,13-trinoreudesm-5-en-7β,8α-diol (1). The structures of isolated compounds were elucidated by extensive spectroscopic methods including 1D and 2D NMR. The structure of compound 2 was confirmed by single-crystal X-ray diffraction analysis.     

Sesquiterpenoids from Inula racemosa

Phytochemical research on the roots of Inula racemosa yielded nine sesquiterpenoids including a new nor-eudesmane-type sesquiterpenoid, 11,12,13-trinoreudesm-5-en-7β,8α-diol (1). The structures of isolated compounds were elucidated by extensive spectroscopic methods including 1D and 2D NMR. The structure of compound 2 was confirmed by single-crystal X-ray diffraction analysis.     

HPLC determination and NMR structural elucidation of sesquiterpene lactones in Inula helenium

A high performance liquid chromatography (HPLC) method was developed for the simultaneous quantification of three bioactive sesquiterpene lactones in Inula helenium L., namely igalane (1), isoalantolactone (2) and alantolactone (3). The HPLC separation was performed on an Agilent Zorbax XDB-C₁₈ column (250 mm × 4.6 mm, 5 μm) with a mobile phase consisting of 55% acetonitrile and 45% water, at a flow rate of 1.0 ml/min, and detected at 210 nm. All three regression equations indicated good linear relationships (r² ≥ 0.9994) between the peak area of each compound and its respective concentration. The HPLC assay was reproducible with overall intra- and inter-day variation of less than 2.85%. The recoveries, measured at three concentrations, varied from 96.60 to 104.43%. This assay was successfully applied to the determination of three bioactive sesquiterpene lactones in eleven samples. The results demonstrated that the assay developed herein was rapid, accurate, reliable and could be readily utilized as a quality control method for I. helenium. In addition, two minor isomers in I. helenium were isolated by semi-preparative HPLC. Their structures were elucidated on the basis of NMR analysis.     

Two new eudesmanolides from Inula racemosa

Two new eudesmane-type sesquiterpene lactones were isolated from the roots of Inula racemosa and their structures were elucidated as 3β-hydroxy-11α,13-dihydroalantolactone (1) and 11α-hydroxy-eudesm-5-en-8β,12-olide (2). Their cytotoxic activities against five human cancer cell lines had been tested and compound 2 exhibited weak cytotoxic activity against BEL-7402 and HCT-8 cell lines. The anti-inflammatory activities were also tested, but neither of them showed any activities.     

Sesquiterpene lactones from Inula hookeri

Four new sesquiterpene lactones, (1 S,5 R,6 S,7 S,8 R,9 R,10 S,11 S)-6-acetoxy-9-hydroxy-4-oxo-pseudoguai-2(3)-en-12,8-olide, (1 S,2 R,5 R,6 S,7 R,8 S,10 R)-6-acetoxy-2-ethoxy-4-oxo-pseudoguai-11(13)-en-12,8-olide, (1 S,2 R,5 R,6 S,7 R,8 S,10 R)-6-acetoxy-2-hydroxy-4-oxo-pseudoguai-11(13)-en-12,8-olide, and 14-acetoxy-1 β,5 α,7 αH-4 β-hydroxy-guai-9(10),11(13)-dien-12,8 α-olide, along with 26 known sesquiterpene lactones, were isolated from the whole plants of Inula hookeri C. B. Clarke. Their structures were established based on spectroscopic methods including HRESIMS, 1D and 2D NMR, and CD techniques. All compounds were evaluated for their cytotoxic activities against HepG2, HeLa, PC-3, and MGC-803 cell lines by CCK-8 assay. Some of the isolates, especiallly pseudoguaianolides and guaianolides, exhibited significant cytotoxicities against these four examined cell lines.     

Cytotoxicity and molecular docking analysis of racemolactone I,a new sesquiterpene lactone isolated from Inula racemosa

ContextTraditionally, Inula racemosa Hook. f. (Asteraceae) has been reported to be effective in cancer treatment which motivated the authors to explore the plant for novel anticancer compounds.ObjectiveTo isolate and characterize new cytotoxic phytoconstituents from I. racemosa roots.Materials and methodsThe column chromatography of I. racemosa ethyl acetate extract furnished a novel sesquiterpene lactone whose structure was established by NMR (1D/2D), ES-MS and its cytotoxic properties were assessed on HeLa, MDAMB-231, and A549 cell lines using MTT and LDH (lactate dehydrogenase) assays. Further, morphological changes were analyzed by flow cytometry, mitochondrial membrane potential, AO-EtBr dual staining, and comet assay. Molecular docking and simulation were performed using Glide and Desmond softwares, respectively, to validate the mechanism of action.ResultsThe isolated compound was identified as racemolactone I (compound 1). Amongst the cell lines tested, considerable changes were observed in HeLa cells. Compound 1 (IC₅₀ = 0.9 µg/mL) significantly decreased cell viability (82%) concomitantly with high LDH release (76%) at 15 µg/mL. Diverse morphological alterations along with significant increase (9.23%) in apoptotic cells and decrease in viable cells were observed. AO-EtBr dual staining also confirmed the presence of 20% apoptotic cells. A gradual decrease in mitochondrial membrane potential was observed. HeLa cells showed significantly increased comet tail length (48.4 µm), indicating broken DNA strands. In silico studies exhibited that compound 1 binds to the active site of Polo-like kinase-1 and forms a stable complex.ConclusionsRacemolactone I was identified as potential anticancer agent, which can further be confirmed by in vivo investigations.     

Cytotoxic sesquiterpene lactones from Eupatorium lindleyanum

Three new germacrane sesquiterpenes, eupalinolides C–E (1–3), along with three known germacrane sesquiterpenes, eupalinolide A (4), eupalinolide B (5), and 3β-acetoxy-8β-(4′-hydroxytigloyloxy)-14-hydroxycostunolide (6), were isolated from Eupatorium lindleyanum. They were tested for cytotoxicity against A-549, BGC-823, SMMC-7721, and HL-60 tumour cell lines. The results showed that these compounds demonstrated potent cytotoxicity. The structures of the compounds were elucidated by means of ¹H and ¹³C NMR spectroscopic analysis, including 2D NMR experiments.     

Cytotoxic sesquiterpene lactones from Eupatorium lindleyanum

Three new germacrane sesquiterpenes, eupalinolides C-E (1-3), along with three known germacrane sesquiterpenes, eupalinolide A (4), eupalinolide B (5), and 3β-acetoxy-8β-(4'-hydroxytigloyloxy)-14-hydroxycostunolide (6), were isolated from Eupatorium lindleyanum. They were tested for cytotoxicity against A-549, BGC-823, SMMC-7721, and HL-60 tumour cell lines. The results showed that these compounds demonstrated potent cytotoxicity. The structures of the compounds were elucidated by means of ¹H and ¹³C NMR spectroscopic analysis, including 2D NMR experiments.