响应面法超声辅助提取优化蛇床子素工艺的研究

对独活中蛇床子素超声提取工艺进行了研究。在单因素实验的基础上,选取了对蛇床子素提取率影响较大的4个因素(温度、乙醇体积分数、提取时间和料液比)。采用响应曲面分析法,建立了独活中蛇床子素提取的二次多项式数学模型,优化蛇床子素的超声提取最佳工艺条件,在温度64.63℃,乙醇体积分数73.18%,提取时间45.33 min,料液比20.67 mL/g条件下蛇床子素最大提取率达到0.987%。表明响应面法超声辅助提取优化蛇床子素工艺的研究方法简便、可靠。     

HPLC法测定蛇床子中蛇床子素的含量

目的:建立蛇床子中蛇床子素的含量测定方法。方法:采用高效液相色谱法,色谱柱选择Agilent ZORBAX SB-C18色谱柱(250 mm×4.6 mm,5μm),流动相为甲醇-体积分数0.05%磷酸溶液(体积比70:30),流速为1.0 mL/min,检测波长为322 nm,柱温30℃。结果:蛇床子素的线性关系方程为Y=4.172×10⁴X+1.912×10²,在12.5~200μg/mL范围内的峰面积与进样浓度线性关系良好(R²=0.9996)。精密度、重复性及稳定性试验的相对标准偏差(RSD)分别为1.28%、1.32%、0.87%,平均加样回收率为99.36%,RSD为1.53%。蛇床子中蛇床子素的平均含量为5.58 mg/g。结论:建立的高效液相色谱法方法简便、准确、快速,重复性好,专属性好,可应用于蛇床子中蛇床子素的含量测定。     

跌打止痛巴布膏体外透皮吸收实验研究

本文采用Franz扩散池和离体裸鼠皮肤进行体外渗透试验,采用HPLC法同时测定蛇床子素和水杨酸甲酯的累积透皮量.以此研究跌打止痛巴布膏体外经皮渗透吸收特征.结果表明两种成分的体外经皮渗透均符合零级动力学方程,蛇床子素和水杨酸甲酯在15 h内的透过率分别为13.5%和49.62%.二者在皮肤的蓄积量分别为17.56%和23.23%.跌打止痛巴布膏中的有效成分在皮肤内有较强的蓄积作用,在15 h内药物持续恒速释放,为控释长效和局部作用的制剂.     

蛇床子炮制历史沿革、化学成分及药理作用研究进展

目的：综述中药蛇床子的炮制概况、化学成分以及药理作用研究进展，为蛇床子不同炮制品的临床合理应用及质量标准研究提供参考依据。方法：借助爱如生古籍库、中国知网、Scifinder等中外文数据库，查阅中药蛇床子的炮制历史沿革、化学成分及药理作用相关文献资料，梳理归纳，并形成综述。结果：古代文献记载的蛇床子炮制方法较多，但现行药品标准多以净制为主；现代研究工艺的研究集中在蛇床子素炮制前后的含量变化；其所含的香豆素类、色原酮类、挥发油类等成分在治疗心血管疾病、抗肿瘤、抗炎、抗骨质疏松、治疗神经性疾病等方面，具有一定药理作用。结论：据记载蛇床子生品有小毒，多外用，但临床也有内服治疗骨质疏松等病症的报道，故不同炮制品的临床应用有待研究；炮制前后的香豆素类成分变化明显，有可能是蛇床子增效减毒的内在机理之一；本综述可为中药蛇床子后续的炮制作用机理、药效物质基础和药理作用研究提供参考。     

蛇床子素通过促进胃癌细胞N87凋亡和细胞周期阻滞而抑制细胞增殖

蛇床子素是从伞形科植物蛇床中提取的一类具有生物活性的化合物。研究显示,蛇床子素对多种肿瘤细胞具有抑制作用,然而尚未有研究揭示其对胃癌N87细胞的抗肿瘤活性。本文研究了蛇床子素在体外和荷瘤小鼠体内对胃癌N87细胞的抗肿瘤效应,并进一步利用流式细胞术、TUNEL试验及Western印迹检测分析其对细胞周期及细胞凋亡的影响,以探索其作用机制。研究结果表明,蛇床子素有效地抑制了体外培养的N87细胞生长,并呈浓度依赖效应。本文还建立了N87的荷瘤小鼠模型。结果显示,无论是在低剂量(50 mg/kg)或高剂量(100 mg/kg)情况下,蛇床子素均显示了有效的肿瘤生长抑制效果。流式细胞术及Western印迹的结果表明,蛇床子素诱导N87细胞阻滞在G_2/M期。通过流式细胞术、TUNEL测试及Western印迹结果证明,蛇床子素通过激活胱天蛋白酶-3依赖的凋亡通路,最终导致了N87细胞凋亡的发生。综上所述,本研究显示,蛇床子素在胃癌N87细胞中通过促进细胞凋亡而发挥其抗肿瘤活性,这将为其应用于胃癌的临床治疗提供理论参考。     

蛇床子素促进人骨肉瘤细胞株SAOS-2凋亡

目的:观察蛇床子素(osthole)对人骨肉瘤细胞SAOS-2增殖和凋亡的影响及潜在的调控机制。方法:采用MTT法、TUNEL染色技术和流式细胞术检测不同浓度蛇床子素对骨肉瘤细胞凋亡的影响;Western blot检测蛇床子素对骨肉瘤细胞中与细胞凋亡密切相关的蛋白(Bax、Bcl-2)的变化。结果:蛇床子素作用于SAOS-2细胞后,MTT结果显示SAOS-2细胞的活力受到明显抑制,且与蛇床子素浓度和时间相关;Western blot结果显示细胞中的促凋亡蛋白Bax表达上调,抗凋亡蛋白Bcl-2表达明显减弱,且呈剂量依赖性。结论:蛇床子素可显著抑制人骨肉瘤细胞的增殖且促进其凋亡的作用,可能与上调凋亡蛋白Bax和下调抗凋亡蛋白Bcl-2的表达有关。     

蛇床子素对离体肺动脉环的作用北大核心CSCD

为了研究蛇床子素对离体肺动脉的作用,本研究游离人及健康SD大鼠的肺动脉血管和SD大鼠的肠系膜动脉血管,剪成长约3 mm的血管环,观察不同浓度的蛇床子素对人和大鼠离体肺动脉以及大鼠肠系膜动脉收缩的作用。结果表明蛇床子素（10-9~10-5mol/L）对人和大鼠苯肾上腺素预收缩的肺动脉具有浓度依赖性的舒张作用,对大鼠肠系膜动脉和正常的肺动脉无舒张作用。证实蛇床子素可舒张人和大鼠离体肺动脉环,其舒张作用具有浓度依赖性和组织特异性。     

蛇床子素对人胶质瘤U251细胞抗增殖作用的研究

目的:研究蛇床子素对人胶质瘤U251细胞的抗增殖作用和可能的机制.方法:不同浓度蛇床子素处理U251细胞后,MTT法检测细胞活力,流式细胞法检测细胞周期和凋亡情况.结果:①蛇床子素显著抑制U251细胞的增殖.②蛇床子素诱导U251细胞发生G2/M期阻滞和凋亡.③蛇床子素处理后的U251细胞PI3K/Akt信号通路活性受到明显抑制.结论:蛇床子素通过抑制PI3K/Akt信号通路抑制人胶质瘤U251细胞的增殖.     

β—环糊精包合荧光法监测蛇床子素的血药浓度

用β-环糊精(β-CD)为包合试剂的荧光法对兔体血浆中蛇床子素的血药浓度进行24h的监测,达峰时间为0.75 h,血中蛇床子素的含量在3.02×10-6～7.26×10-6g/ml之间,方法简便、快速、效果良好.     

蛇床子植物学相关研究进展

蛇床子是一种传统中药,是伞形科（Apiaceae）植物蛇床（Cnidium monnieri）的干燥成熟果实,主要有效成分为蛇床子素及其它香豆素类化合物。蛇床子具有抗心律失常、抗病毒、抗肿瘤等多种功效,并且广泛应用于白癜风、银屑病等多种皮肤病的治疗。蛇床适应性强,分布广泛,但不同产地的蛇床子药材品质差异较大,需要仔细鉴别入药。对蛇床的生物学特性、产地特征、栽培研究和鉴别方法等进行了综述,并展望了未来产学研结合的蛇床子植物学研究的发展趋势。     

An efficient strategy based on MAE, HPLC-DAD-ESI-MS/MS and 2D-prep-HPLC-DAD for the rapid extraction, separation, identification and purification of five active coumarin components from Radix Angelicae Dahuricae

Introduction – Further studies of active coumarin components in Radix Angelicae Dahuricae (AE) are absolutely essential to provide data on pharmacology, toxicology and quality for innovative drug candidates. Thus, the preparation of active component standards and the administration of coumarin monomers should be carried out. The isolation of the low‐level active components from complex Traditional Chinese Medicine (TCM) samples necessitates the development of rapid, simple and economical modern extraction, separation, identification and purification methods. Objective – To develop an efficient strategy for the rapid extraction, separation, identification and purification of coumarins from AE. Methodology – First, active coumarins in AE were extracted with microwave‐assisted extraction (MAE) after the extraction conditions were optimised. Second, gradient extraction methods with MAE were used to partially purify AE. Third, a high‐performance liquid chromatography–diode array detection‐electrospray ionisation tandem mass spectrometry (HPLC‐DAD‐ESI‐MS/MS) method was applied for the preliminary on‐line identification and screening of the main coumarins in AE extract. Finally, a two‐dimensional preparative high‐performance liquid chromatography–diode array detection (2D‐prep‐HPLC‐DAD) system was developed for further preparative separation of those target components. Results – Altogether 10 coumarins have been identified and five of them including xanthotoxol, osthenol, oxypeucedanin hydrate, byakangelicin and imperatorin were deemed as target components for the preparative isolation. All of the five isolated coumarins were at high purities of over 99% and the production rate was much higher than the traditional methods. Conclusion – The present paper demonstrates that these consecutive approaches are very useful for to isolate chemical constituents from TCM.     

福建野生单叶蔓荆子总黄酮与蔓荆子黄素含量分析

对福建省不同地区野生单叶蔓荆子总黄酮和蔓荆子黄素含量进行测定,不同地区含量差异进行比较分析,确定福建野生单叶蔓荆子的质量水平,为进一步开发福建野生单叶蔓荆子提供基础研究数据。回流提取总黄酮,紫外分光光度法(UV法)测定总黄酮含量,高效液相色谱法(HPLC法)测定蔓荆子黄素含量。结果表明不同产地野生单叶蔓荆子总黄酮含量范围为7.081%～10.792%,平均含量9.040%;蔓荆子黄素含量范围为0.06170%～0.12578%,平均含量0.08878%。福建省不同地区野生单叶蔓荆子质量(以蔓荆子黄素含量为指标)均符合药典规定,可进一步开发利用。     

Influence of the extraction mode on the yield of hyperoside, vitexin and vitexin-2'-O-rhamnoside from Crataegus monogyna Jacq. (hawthorn)

Introduction –  The extract of Crataegus monogyna shows sedative, hypotensive, vasodilator and cardio‐tonic actions. Although several papers dealing with the extraction of metabolites from Crataegus have been published, the plant productivity in terms of bioactive compounds is not easily understandable as yet. Objective –  To investigate the influence of the extraction mode on the yield of bioactive compounds from Crataegus monogyna Jacq. in order to evaluate plant productivity. Methodology –  Samples were prepared by extraction of powdered material obtained from top branches, flowers and leaves. Soxhlet extraction, maceration and ultrasound‐ and microwave‐assisted extraction at different experimental conditions were investigated for the exhaustive extraction of hyperoside, vitexin and vitexin‐2′′‐O‐rhamnoside. The phytocomponents were identified and quantified by HPLC‐UV/PAD, comparing HPLC retention times and UV spectra of individual peaks with those of the standards analysed under the same conditions. Results –  An easy‐to‐use HPLC isocratic method suitable for the quantification of hyperoside, vitexin and vitexin‐2′′‐O‐rhamnoside in raw plant extracts was developed. The optimised HPLC methodology was applied to evaluate different extraction procedures. The ultrasound and microwave‐assisted extraction protocols showed higher extraction efficiency than the others. In particular, the optimised microwave protocol gave rise to the highest extraction efficiency with high reproducibility. Conclusions –  A microwave protocol combined with isocratic HPLC analysis is proposed for the rapid screening of plant materials collected in different environmental conditions in order to evaluate the productivity of Crataegus monogyna Jacq. and to find out the best ecological conditions to cultivate hawthorn in Northern Italy. Copyright © 2008 John Wiley & Sons, Ltd.     

Cytohistological and phytochemical study of madder root extracts obtained by ultrasonic and classical extractions

Introduction – Madder (Rubia tinctorum) has been used since ancient times as a source of pigments for dyeing and painting. Madder dyes are localised in roots and the native chemical population is composed of glycosiled and aglycone compounds. The aim of this study is to elaborate an efficient extraction process without any chemical denaturation of dyes. Objective – To compare an optimised ultrasonic process, using for madder dye extraction, with two conventional procedures and to determine the efficiency of ultrasound on these vegetable matrix. Methodology – Madder roots were extract in a methanol–water mixture in 37 : 63 (v/v) for ultrasound and 80 : 20 (v/v) for reflux and agitation. HPLC‐PAD analyses showed the anthraquinone proportion for each extraction process and their denaturing effects. Finally, cytohistological observations were made to show the consequence of each process on the cell organisation in madder roots. Results – The results showed that the amount of extracted dyes was higher with UAE than with agitation and reflux. HPLC‐PAD analysis revealed that the anthraquinone composition differed according to the extraction procedure. The UAE extracts presented an important richness in terms of anthraquinonic compounds that suggests a preserving effect. Cytohistological observations showed that the main alterations concerned the cell walls of phloem. After UAE the walls exhibited numerous pitted areas reflecting an ultrasound‐induced cavitation that enhances the extraction effectiveness of this method. Conclusion – The study has shown the improvement of madder roots extraction both quantitatively and qualitatively using the efficiency of ultrasound‐assisted extraction in comparison with magnetic agitation and reflux techniques. Copyright © 2009 John Wiley & Sons, Ltd.     

HPLC comparison of supercritical fluid extraction and solvent extraction of coumarins from the peel of Citrus maxima fruit

The efficiency of carbon dioxide supercritical fluid extraction (SFE) of the biologically active compounds imperatorin, meranzin and meranzin hydrate from the fruit peel of Citrus maxima Merr. has been compared with that of solvent extraction with acetone. Under the best SFE conditions tested for the three coumarins, which involved extraction at 50 degrees C and 27.6 MPa, the extractive efficiencies were 84, 76 and 18% for imperatorin, meranzin and meranzin hydrate, respectively. The presence of modifiers significantly affected the extraction efficiency: the highest extraction efficiency of the three coumarins was obtained with ethanol as modifier.     

Computer-assisted, high-performance liquid chromatography with mass spectrometric detection for the analysis of coumarins in Peucedanum palustre and Angelica archangelica

A reversed-phase HPLC method with atmospheric pressure chemical ionisation MS detection has been developed for the separation and identification of coumarins in plants of Peucedanum palustre L. (Moench) and Angelica archangelica (L.) var. archangelica. The Turbo Method Development program was utilised to optimise the mobile phase with two organic solvents (acetonitrile and methanol) and two aqueous solutions (1.0% formic acid and 10 mM ammonium acetate). Optimisation of the solvent gradients for the method was performed with the aid of the DryLab program. Analyses were carried out using a Phenomenex Prodigy RP C18 column. Fifty-two peaks (14 of which were associated with coumarins) were separated in 30 min from extracts of P. palustre, and 48 peaks (15 associated with coumarins) from extracts of A. archangelica. A total of 21 different coumarin-type compounds were identified in the aerial and the underground parts of the title plants. Isopimpinellin and pimpinellin were found for the first time in P. palustre and were identified by comparison of retention times and MS data obtained following the analysis of pure standards. This is the first report of the coumarin composition of the umbels of P. palustre.     

重齿当归根中二种香豆素含量的HPLC法测定

重齿当归〔Ａｎｇｅｌｉｃａｂｉｓｅｒｒａｔａ (ＳｈａｎｅｔＹｕａｎ)ＹｕａｎｅｔＳｈａｎ〕系中药独活原植物 ,又名重齿毛当归 ,是我国特产的常用中药 ,有祛风、除湿、散寒、止痛等功效。四川、湖北、陕西等省都有栽培。野生重齿当归广泛分布于浙江、安徽、江西、四川等省     

Simultaneous determination of theophylline and its major metabolites in urine by reversed-phase ion-pair high-performance liquid chromatography

A new, highly selective high-performance liquid-chromatographic (HPLC) assay for theophylline and its major metabolites in urine is described. The method utilizes an ion-pair extraction followed by separation and quantitation by reversed-phase ion-pair gradient-elution HPLC. Comparison with several other methods showed that interferences were present in too many blank urine samples to allow for the accurate quantitation of the metabolites of theophylline by direct injection-isocratic HPLC assays. Sample processing involving ion-pair complexing and extraction together with gradient-elution systems is recommended for accurate pharmacokinetic studies.     

Coumarins from Musineon divaricatum

A chemical investigation of Musineon divaricatum has resulted in the isolation of 19 coumarins, five of which are novel compounds; 18 are khellactone derivatives. The coumarins were isolated from the crude extracts by a combination of adsorption chromatography, gel permeation and HPLC; the more successful HPLC separations utilized a nitrile bonded phase column. The structures were determined by ¹H NMR and mass spectral studies and by comparisons with literature data. The relative configuration for the entire series was secured from ¹H NMR data, while the absolute configuration could be assigned with any certainty only in the cases of two of the compounds.     

Extraction of pigments and fatty acids from the green alga Scenedesmus obliquus (Chlorophyceae)

In this paper, the efficiency of pigment and fatty acid extraction from resistant algae using Scenedesmus obliquus as an example was examined. We found that adding quartz sand and solvent to freeze-dried algal material and subsequent extraction in an ultrasound bath for 90min at –4°C resulted in excellent extraction of these compounds. This extraction method was compared with a method regularly used for extraction of fatty acids and pigments, i.e. addition of solvents to algal material with subsequent incubation. Our extraction using the ultrasound and sand method was about twice as efficient as this method for both pigments and fatty acids. The ultrasound method is simple, extracts over 90% of the different substances in one step and conserves the relationships of pigments and fatty acids. In addition, no alteration- or breakdown products were observed with the new method. Thus, this method allows accurate quantitative extraction of both pigments and fatty acids from Scenedesmus obliquus and other algae. The method was also been found to be as effective for Cryptomonas erosa (Cryptophyceae), Cyclotella meneghiniana (Bacillariophyceae), Microcystis aeruginosa (Cyanophyceae), and Staurastrum paradoxum (Chlorophyceae, Desmidiaceae) and is thus applicable to a wide spectrum of algae.