原阿片碱提取工艺的改进

采用酸碱法处理从夏天无中提取原阿片碱,所得产品理化性质与标准品一致。     

东北延胡索愈伤组织培养及延胡索乙素形成的适宜条件

比较了东北产两种延胡索各器官的愈伤组织发生情况,采用单因子和正交设计试验,对齿瓣延胡索(Corydalis remota)愈伤组织生长及延胡索乙素形成的适宜条件做了研究。结果表明,齿瓣延胡索各器官对培养基的响应性均显著高于东北延胡索(C. ambigua),其块茎愈伤诱导率高达93.3%;MS培养基对于愈伤组织诱导较White有利, 2 mg·L^-1 2, 4-D与1 mg·L^-1 KT的组合愈伤诱导率最高,并适于继代培养。正交试验结果表明,愈伤组织生长和延胡索乙素形成的适宜条件存在差异,在27组处理中, 1号培养基有利于培养物鲜重的提高, 9号有利于干物质的积累, 10号则有利于的延胡索乙素形成;培养基中几种无机离子和有机成分均显著影响愈伤组织生长和延胡索乙素形成,NH₄⁺/NO₃^-与Ca²⁺和K⁺之间的交互作用对于生物量积累和延胡索乙素含量均有显著影响。     

培养条件对齿瓣延胡索愈伤组织生长及延胡索乙素含量的影响

研究了不同培养条件对齿瓣延胡索(Corydalis remota)愈伤组织生长及延胡索乙素(THP)含量的影响。结果表明, 2, 4-D较NAA更有利于提高培养物鲜重,加速细胞增殖。培养基添加2.0 mg/L 2, 4-D和1.0 mg/L BA不仅有利于干物质积累,而且显著提高THP水平。蔗糖不仅可提高培养物鲜重,而且与半乳糖组合可显著促进干物质积累和THP水平的提高。水解酪蛋白和水解乳蛋白均有利于愈伤组织生长和THP含量的提高, 450mg/L的水解乳蛋白处理组THP含量最高。光照可提高培养物鲜重和THP含量。     

延胡索乙素与白芷总香豆素、总挥发油配伍药效学比较研究

目的：评价延胡索乙素、白芷总香豆素及总挥发油配伍的药效学比较研究。方法：小鼠醋酸扭体法、热板镇痛法、甩尾法三种模型比较元胡止痛片（A组）、延胡索乙素（B组）、白芷总香豆素（C组）、白芷总挥发油（D组）、延胡索乙素与白芷总香豆素配伍（E组）、延胡索乙素与白芷总挥发油配伍（F组）及延胡索乙素与白芷总香豆素、白芷总挥发油配伍（G组）后的镇痛作用。结果：A、G组镇痛效果最佳,B、E、F组具显著的镇痛作用,C、D组与模型组相比,有一定的镇痛作用,但较弱。结论：延胡索乙素与白芷总香豆素、总挥发油配伍使用,具有显著的镇痛作用,类似于元胡止痛片中元胡白芷的配伍作用,但成分更为明确,标准可控,使用量小,属于分子中药组方范畴。元胡的主要有效成分延胡索乙素与白芷的主要成分白芷总香豆素、总挥发油均具有镇痛作用,符合中药元胡及白芷的功效,本实验采用药物有效成分进行配伍,成分明确,结果可控。     

中药延胡索乙素对家兔心脏舒张期与收缩期比值的影响

目的：探索中药成分延胡索乙素对家兔心脏舒张期与收缩期比值的影响。方法：用家兔作为实验动物,选用中药成分延胡索乙素作实验药物并用生理盐水作对照,观察静脉注射后家兔心脏舒张期与收缩期比值（D/S）的变化。结果：静脉注射延胡索乙素具有降心率和升高D/S比值的效应,1周后重复实验得到类似结果,而生理盐水没有这种作用。结论：延胡索乙素具有降心率和升高D/S比值的作用,但延胡索乙素升D/S比值的临床应用还需要进一步研究。     

土壤有效元素含量与延胡索(Corydalis yanhusuo)品质的相关性研究

延胡索乙素是相关延胡索(Corydalis yanhusuo)品质的重要化学成分,在中国药典中作为延胡索质量控制指标,分析不同产地延胡索中延胡索乙素含量与根际土壤有效性元素含量的相关性,对指导延胡索合理化种植栽培有重要科学意义.采用土壤农化法对不同产地延胡索根际土壤中10种元素(大量元素N、P、K、Ca、Mg;微量元素...     

规范化种植延胡索药材的质量评价

通过对江西红壤丘陵坡地规范化种植的延胡索（Corydalis yanhusuo W.T.Wang）中延胡索乙素、重金属和农残连续4年跟踪检测分析,对延胡索的药效成分和安全性进行质量评价。延胡索乙素含量采用高效液相法测定,重金属含量采用原子吸收分光光度法测定,农药残留采用气相色谱法测定。结果表明：电视延胡索乙素含量为0.066%~0.088%,重金属铅0.76mg/kg、镉0.22mg/kg、砷0.0025mg/kg、汞0.007mg/kg、铜5.26mg/kg、有机氯农药六六六0.033mg/kg,滴滴涕和五氯硝基苯为痕迹量。延胡索新种植区---江西红壤丘陵坡地规范化种植的延胡索符合《中华人民共和国药典》2010年版一部及《药用植物及制剂外经贸委绿色行业标准》WM/T2-2004的规定。     

从千层塔中微波协助提取石杉碱甲和石杉碱乙

首先从 8种溶剂中筛选出硫酸作为微波协助提取的浸提溶剂 ,然后用正交试验确立了千层塔生物碱的最佳提取条件。以石杉碱甲和石杉碱乙回收率为指标 ,考察了溶剂倍数、溶剂浓度、微波处理时间等因素。结果表明 ,在室温下微波协助提取的最优条件为 :酸浓度 0.8% (v/v) ,液固比例 2 5∶1 ,微波处理时间 90s。 3次重复实验所得石杉碱甲和石杉碱乙回收率分别是 93.7%和 93.9% ,相对标准偏差分别为 1.79%和 1.5 6% (n =3)。与传统的回流提取工艺相比 ,过程时间从 2h缩短为 90s,回收率提高了 1 0 %以上。     

HPLC法检测蛇足石杉内生真菌胶胞炭疽发酵液中石杉碱甲和石杉碱乙的含量

建立了高效液相色谱(HPLC)测定蛇足石杉(Huperzia serrate)内生真菌胶胞炭疽发酵液中石杉碱甲(Huperzina A)和石杉碱乙(Huperzine B)含量的方法,并以此方法检测胶胞炭疽发酵液中石杉碱甲和石杉碱乙含量的积累。内生真菌发酵液经氯仿萃取、甲醇溶解、过滤后进行高效液相检测分析,选用Agilent Eclipse plus-C18色谱柱(250 mm×4.6 mm,5μm),以0.015 mol/L乙酸铵(p H 6.8)和甲醇溶液(70∶30)为流动相进行等度洗脱,流速1 mL/min,检测波长为308 nm,连续检测内生真菌胶胞炭疽发酵液中第6–15天石杉碱甲和石杉碱乙的含量积累。结果表明,发酵提取液中的石杉碱甲和石杉碱乙可在25 min内进行很好的分离和分析,石杉碱甲在1.50-48.00μg/mL范围内线性关系良好(相关系数r为0.999 5),石杉碱乙在0.25-7.50μg/mL范围内线性关系良好(相关系数r为0.999 7),石杉碱甲和石杉碱乙的平均加标回收率分别为106.83%、108.06%,相对标准偏差(RSD)分别为3.34%、3.60%。该方法简便、快速、精密度高、结果准确,适用于内生真菌发酵液中石杉碱甲和石杉碱乙含量检测。在发酵过程中,内生真菌发酵液中石杉碱甲和石杉碱乙的含量呈现先增后减,随后有所增加继而又减少的趋势。石杉碱甲和石杉碱乙的含量分别在内生真菌发酵第14天、第8天达到最高,分别为12.417 0μg/mL、4.660 3μg/mL。该方法学的建立为内生真菌胶胞炭疽合成石杉碱甲与石杉碱乙的机制研究提供了检测手段,从而有利于药物新资源的开发。     

野甘菊中小白菊内酯的超临界流体色谱测定

用超临界流体色谱法分析野甘菊中小白菊内酯时 ,具有很好的精密度和线性关系 ,其保留时间和峰面积的相对标准偏差分别为 2 6 7%和 0 70 % ,相关系数为 0 9996。样品经甲醇提取后直接进样分析 ,并在 3min内完成 ,可用于快速检测     

延胡索的化学成分研究(英文)

采用现代分离技术和方法,从延胡索根茎中分离得到17个化合物,通过波谱分析鉴定其结构。包括11个生物碱类化合物,二氢白屈菜红碱(1)、去氢紫堇碱(2)、四氢非洲防己胺(3)、异紫堇球碱(4)、紫堇碱(5)、四氢黄连碱(6)、药根碱(7)、黄连碱(8)、小檗碱(12)、巴马汀(13)和延胡索乙素(14);2个蒽醌类化合物,大黄素(9)和大黄素甲醚(10);1个三萜类化合物,即3β-羟基-齐墩果烷-111,3(18)-二烯-28-酸(11)和3个甾醇类化合物,豆甾醇(15)、β-谷甾醇(16)和胡萝卜苷(17)。其中,化合物9～11首次从该属植物中分离得到,1首次从该种中分离得到。     

夏天无中普鲁托品高效液相含量测定提取方法的研究

采用RP -HPLC测定了夏天无中普鲁托品的含量 ,比较了超声时间、提取溶剂、加热对夏天无中普鲁托品含量测定的影响 ,结果表明 :加热后超声法对普鲁托品的提取比普通超声提取法高 30 %以上     

GC-MS技术在延胡索块茎代谢产物研究中的应用

采用气相色谱-质谱联用(GC-MS)技术建立了延胡索(Corydalis yanhusuo)块茎形成过程中代谢产物的代谢组学研究方法。考察了不同体积提取液和衍生化试剂MSTFA对峰的信号强度和个数的影响, 优化了实验条件, 确定每100 mg材料用300 μL提取液与100 μL衍生化试剂MSTFA时, 效果最佳。对延胡索块茎样品进行测定, 结果共得到92种化合物, 分别为: 5种脂肪酸、21种有机酸、16种氨基酸、6种糖类、23种其它成分和21种未知化合物; 并且利用主成分分析(PCA)数据处理方法对延胡索块茎代谢产物进行了模式识别研究。结果表明: 该实验方法稳定、可靠, 且在主成分分析模式下其识别结果有明显的分类, 因此可为延胡索块茎的进一步研究提供有效数据, 并为相关研究提供参考。     

Enantiomeric Separation of Original Heterocyclic Organophosphorus Compounds in Supercritical Fluid Chromatography

New and original heterocyclic α-enamido phosphine chiral solutes were prepared: four structurally similar racemates with the chirality center placed on the phosphorus atom, and four other related pairs of enantiomers with chirality borne by the carbon atoms of the phospholane ring. The structural variations were placed on an aliphatic heterocycle (six- or seven-member rings) and on the carbamate function (methyl or t-butyl). Their separation was achieved on a commercial cellulose tris-(3,5-dimethylphenylcarbamate) stationary phase (Lux Cellulose-1, Phenomenex) in supercritical fluid chromatography (SFC). The effects of molecular structure on SFC retention and enantioresolution were studied. Among these eight pairs of enantiomers, some reversal of elution order between similar compounds was observed. The effect of changing the organic solvent (methanol and ethanol) and its proportion (between 5 and 40%) in the mobile phase was investigated. Retention data were collected over the temperature range 0–50 °C, and the results interpreted from thermodynamic aspects. Chirality, 2013. © 2013 Wiley Periodicals, Inc.     

Effects of Hexane in Supercritical Fluid Chromatography for the Separation of Enantiomers

Supercritical fluid chromatography (SFC), operated in conventional mode, is normally recognized as normal phase chromatography, and uses a solvent combination of supercritical CO₂ and alcohols to separate compounds. Hexane, a commonly used solvent in normal phase liquid chromatography (NP‐LC), is rarely used in SFC and, in some cases, is added to the organic modifiers to increase liquid content in order to achieve better efficiency in preparative SFC for poorly retained compounds. Although hexane is believed to have similar solvent strength to that of supercritical CO₂, its effects on the enantioseparation in SFC is largely unknown. To understand the chromatographic effects of an apolar solvent, such as hexane in SFC, we compared the chromatographic behaviors of 35 chiral compounds using a parallel SFC method under traditional SFC mode of only “pure” alcohol‐CO₂ to that of hexane‐assisted SFC (HA‐SFC), which uses mixtures of alcohol and hexane (as cosolvents) and CO₂. We observed that, in some cases, hexane behaves just like supercritical CO₂, where replacement of a portion of CO₂ with hexane does not significantly change retention times or resolution of the peaks. In many cases, however, addition of hexane in mobile phases does affect chromatographic behavior of one or both enantiomers. Such effects might provide opportunities for separation of some enantiomers. Chirality 28:192–198, 2016. © 2016 Wiley Periodicals, Inc.     

正交设计优化微波辅助测定延胡索叶总生物碱含量

通过微波辅助提取的方式,以延胡索叶总生物碱含量为考察指标,通过单因素试验和正交试验对总生物碱提取方法进行优化。重点考察了微波提取温度、时间、料液比、提取缓冲液pH值等因素对延胡索叶总生物碱得率的影响。各因素对延胡索叶总生物碱提取量的影响大小依次为:提取缓冲液pH值料液比提取温度。其最佳的提取方法为:提取缓冲液pH值为1. 0,料液比1∶100,提取温度60℃,提取时间20 min。在此条件下,叶总生物碱含量为25. 910 mg/g。利用微波辅助提取延胡索叶中总生物碱,不仅耗时短,效率高,而且操作简单、稳定,为延胡索叶总生物碱含量测定提供一定的技术支持。     

Simultaneous Measurement of Fluoroquinolones in Eggs by a Combination of Supercritical Fluid Extraction and High Pressure Liquid Chromatography

Simultaneous detection of the fluoroquinolone antibiotics ciprofloxacin, enrofloxacin, ofloxacin, and norfloxacin in eggs by a combination of supercritical fluid extraction (SFE) and high pressure liquid chromatography (HPLC) was studied. Lipid matrices that have been considered to result in poor extraction and isolation of fluoroquinolones in eggs were removed first by SFE with supercritical CO₂ alone, and then the fluoroquinolones were extracted by SFE with supercritical CO₂ containing 20% (v/v) methanol for HPLC analysis. A time-course study of the extraction of lipid matrices of eggs suggested that the SFE method successfully removed the matrices within 20 min. When the fluoroquinolones added to control eggs were extracted by SFE, the extraction efficiency was similar to that by the solvent extraction method, giving the recovery percentages from 83 to 96% in a 40 min-extraction time. The fluoroquinolones extracted from eggs by SFE were analyzed simultaneously by HPLC equipped with a fluorescence detector with detection sensitivity at about 10 ppb for the detection limit. The standard calibration profiles of fluoroquinolones showed linear responses to HPLC, showing more than 0.995 for the mean r ² value. This is the first report of the simultaneous measurement of fluoroquinolones in eggs by a combination of SFE and HPLC. Using the SFE method allowed us to avoid extensive sample preparation such as solvent extraction and chromatographic cleanup that are basically required in extraction of fluoroquinolones.     

The Effect of Armillaria mellea Elicitor on Alkaloid Production in Tissue Cultures of Corydalis yanhusuo

It has shown that alkaloid accumulation in tissue cultures of Corydalis yanhusuo orchestrated to some extent the growth in cultures. Some alkaloids, such as tetrahyclropalmatine continued to increase in the static phase of culture growth. The culture solution of Armillaria mellea as an elicitor was able to promote alkaloids accumulation in cultures, as, for example, protopine was increased up to 0.114% DW. by the eliciting but remained at trace level in cultures without elicitor.     

Chiral Separation of G‐type Chemical Warfare Nerve Agents via Analytical Supercritical Fluid Chromatography

Chemical warfare nerve agents (CWNAs) are extremely toxic organophosphorus compounds that contain a chiral phosphorus center. Undirected synthesis of G‐type CWNAs produces stereoisomers of tabun, sarin, soman, and cyclosarin (GA, GB, GD, and GF, respectively). Analytical‐scale methods were developed using a supercritical fluid chromatography (SFC) system in tandem with a mass spectrometer for the separation, quantitation, and isolation of individual stereoisomers of GA, GB, GD, and GF. Screening various chiral stationary phases (CSPs) for the capacity to provide full baseline separation of the CWNAs revealed that a Regis WhelkO1 (SS) column was capable of separating the enantiomers of GA, GB, and GF, with elution of the P(+) enantiomer preceding elution of the corresponding P(–) enantiomer; two WhelkO1 (SS) columns had to be connected in series to achieve complete baseline resolution. The four diastereomers of GD were also resolved using two tandem WhelkO1 (SS) columns, with complete baseline separation of the two P(+) epimers. A single WhelkO1 (RR) column with inverse stereochemistry resulted in baseline separation of the GD P(–) epimers. The analytical methods described can be scaled to allow isolation of individual stereoisomers to assist in screening and development of countermeasures to organophosphorus nerve agents. Chirality 26:817–824, 2014. © 2014 The Authors. Chirality published by John Wiley Periodicals, Inc.