青枯菌HPLC分析中样品制备方法的优化

本文研究了青枯菌细胞的制备方法对细胞生命活力和表面特性的影响。结果表明,在高效离子交换色谱分析（HPLC）中,采用5000×g离心10min收集菌体细胞、超纯水（＞16MΩ）悬浮和洗涤青枯菌、重复洗涤二次的制备方法,既可以避免培养基成分造成的干扰,又可以保持青枯菌细胞的生命活力和细胞表面的原有性质。     

柘荣太子参HPLC指纹图谱的研究

建立太子参的 HPLC指纹图谱分析条件,为太子参药材内在质量评价积累数据.方法：应用RP-HPLC法;Cosmosil C18分析柱;乙腈-水二元梯度洗脱;流速为1.0 mL/in;检测波长203 nm;分析时间60 min.结果：建立太子参药材指纹图谱,特征共有峰有15个.结论：该方法准确可靠,重复性好,可用于太子参的HPLC的指纹图谱分析.     

野生多叶棘豆中芦丁的动态积累规律

目的：为确定野生多叶棘豆的最佳采收期提供实验依据.方法：采用高效液相色谱法对不同采收期野生多叶棘豆中芦丁含量进行测定,考察其指标成分芦丁的含量变化趋势.结果：多叶棘豆中芦丁的动态积累有一定规律.在7月末8月初以前其芦丁含量有上升趋势,8月初以后其芦丁含量逐渐下降.结论：为保证药材的质量,根据多叶棘豆中有效成分的动态积累规律,有必要在其最佳采收期采集药材.     

固相小柱对沙苑子总黄酮萃取方法的研究

目的:以沙苑子总黄酮为研究对象,考察固相萃取小柱的洗脱条件,确定固相萃取沙苑子总黄酮的实验方法。方法:以不同浓度的甲醇对不同填料的固相小柱进行洗脱,将洗脱液在岛津VP-ODS(150 mm×4.6 mm,5μm)柱上进行高效液相色谱分析。通过比较不同浓度及体积甲醇的萃取效率,确定沙苑子苷在固相萃取小柱上的洗脱条件。结果:用正相氨基小柱以50%甲醇洗脱效果好,回收率可达100%。结论:经试验,正相氨基小柱对沙苑子苷的萃取回收率约为100%,适用于沙苑子总黄酮中沙苑子苷的分析。     

Valepotriates accumulation in callus, suspended cells and untransformed root cultures of Valeriana glechomifolia

Summary Valeriana glechomifolia is an endemic species of southern Brazil, capable of accumulating, in all of its organs, the terpene derivatives known as valepotriates, the presumed sedative components of the roots of pharmaceutically used species of Valeriana. In vitro cultures of the plant were established and the accumulation of acevaltrate, didrovaltrate, and valtrate in callus, cell suspension, and untransformed root cultures was studied. Leaves of in natura plants and roots of micropropagated plantlets were used as the explants for callus induction and root culture establishment, respectively, on Gamborg B5 basal medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) alone or with kinetin (KIN). Culture growth and secondary metabolite yields were enhanced with 2,4-D (4.52μM) and KIN (0.93μM). Maximum valepotriate contents, quantified by HPLC, of acevaltrate (ACE) 2.6mg g⁻¹ DW, valtrate (VAL) 10.2mgg⁻¹ DW, and didrovaltrate (DID) 2.9mg g⁻¹ DW were observed in root cultures after 7–8wk of culture.     

Utilization of mustard waste isolates for improved production of astaxanthin by Xanthophyllomyces dendrorhous

Astaxanthin production in the wild strain Xanthophyllomyces dendrorhous TISTR 5730 was investigated using different mustard waste media, including mustard waste residue extract (MRE), mustard waste residue hydrolysate (MRH), mustard waste precipitated extract (MPE), and mustard waste precipitated hydrolysate (MPH). The growth of X. dendrorhous and the production of astaxanthin were dependent on the type and initial concentrations of mustard waste media. The MPH medium was the best substrate resulting in yields of biomass and astaxanthin of 19.6 g/L and 25.8 mg/L, respectively, under optimal conditions. MPH medium improved astaxanthin production 11-fold compared to the commonly used commercial yeast malt medium, and 1.3–2.1-fold compared to other mustard waste media.     

基于HPLC-ECD研究牡荆叶抗氧化的谱-效关系

为研究牡荆叶指纹图谱与抗氧化活性的谱-效关系，该研究首先建立了18批牡荆叶的高效液相色谱-电化学检测法(HPLC-ECD)指纹图谱，对不同来源牡荆叶药材进行聚类分析，鉴定主要酚类化合物且测定其含量，分析牡荆叶的总酚和总黄酮含量，并采用DPPH自由基清除法、ABTS自由基清除法、氧自由基吸收能力法及铁离子还原能力法考察其体外抗氧化活性，通过皮尔逊相关分析、灰度关联分析及偏最小二乘回归分析法研究牡荆叶的谱-效关系。结果表明：(1)牡荆叶的指纹图谱标定21个共有峰，共指认出10个峰，其含量顺序为绿原酸>异荭草苷>木犀草苷>异牡荆素>异绿原酸A>异绿原酸C>原儿茶酸>荭草苷>异绿原酸B>新绿原酸；不同产地样品间相似性较高，相似度结果为0.816～0.983。(2)系统聚类分析显示，样品含量对分类有一定影响，不同来源样品被分为3类，其中南北方样品存在一定差异。(3)牡荆叶中总酚和总黄酮的含量分别为15.82～61.83 mg·g^-1和27.85～157.65 mg·g^-1,样品均具不同程度的抗氧化活...     

Comparison of three different extraction methods and HPLC determination of the anthraquinones aloe‐emodine,emodine, rheine,chrysophanol and physcione in the bark of Rhamnus alpinus L. (Rhamnaceae)

Introduction – Rhamnus alpinus L. (Rhamnaceae), a traditional plants in the flora of the Abruzzo region, is known to contain active anthraquinone secondary metabolites. However, the content of anthraquinones varies among R. alpinus samples depending on collection season and site. Thus, using simple, reliable and accurate analytical methods for the determination of anthraquinones in R. alpinus extracts allows comparative study of different methods of extraction. Objective – After a partial validation of an HPLC method for the simultaneous determination of five anthraquinones, aloe‐emodine, rheine, emodine, chrysophanol and physcione, in the bark of R. alpinus, we compared three different methods of extraction. Methodology – Anthraquinones were extracted from the bark of R. alpinus using different techniques (methanol maceration, ultrasonic and supercritical CO₂ extraction). Separation and quantification of anthraquinones were accomplished using a reversed‐phase C₁₈ column with the mobile phase of H₂O–methanol (40 : 60, v/v, 1% formic acid) at a wavelength of 254 nm. The qualitative analyses were also achieved at wavelength of 435 nm. Results – All calibration curves were linear over the concentration range tested (10–200 mM) with the determination coefficients ≥0.991. The detection limits (S/N = 3) were 5 mM for each analytes. All five anthraquinones were found in the samples tested at concentrations reported in experimental data. Conclusion – The described HPLC method and optimised extraction procedure are simple, accurate and selective for separation and quantification of anthraquinones in the bark of R. alpinus and allow evaluation of the best extraction procedure between the tested assays. Copyright © 2009 John Wiley & Sons, Ltd.     

Production of leukotriene B4 and prostaglandin E2 by turbot (Scophthalmus maximus) leukocytes

Production of two eicosanoids derived from lipoxygenase and cyclooxygenase activities: leukotriene B₄ (LTB₄) and prostaglandin E₂ (PGE₂), respectively, have been simultaneously determined in turbot (Scophthalmus maximus) blood leucocyte and kidney macrophage supernatants by a reverse phase high performance liquid chromatography (HPLC) system coupled with a Diode–Array detector. Levels of LTB₄ after calcium ionophore challenge were 4.08 ng ml⁻¹ in blood leukocyte supernatants and 0.25 ng ml⁻¹ in kidney macrophage supernatants. The levels found for PGE₂ were 428.23 and 606.67 ng ml⁻¹ in blood leukocytes and kidney macrophage supernatants, respectively. When blood leukocytes were treated with the respective inhibitors for the enzymes implicated on the synthesis of both compounds an inhibition of 90.35% was observed for PGE₂ and 76.44% for LTB₄. The detection limit of the method was 0.15 ng ml⁻¹ for LTB₄ and 50 ng ml⁻¹ for PGE₂.