高效液相色谱-串联质谱法同时测定马鞭草中5种糖苷类成分的含量

目的建立高效液相色谱-串联质谱(HPLC-MS/MS)法同时测定马鞭草中桃叶珊瑚苷、戟叶马鞭草苷、马鞭草苷、龙胆苦苷、毛蕊花糖苷的含量。方法色谱柱采用Waters SunfireTMC18(4.6 mm×150 mm,5μm),流动相为乙腈-0.1%甲酸水(25∶75→40∶60),梯度洗脱,流速0.8 m L/min,进样量5μL,柱温30℃;采用电喷雾离子源(ESI),以多反应监测方式(MRM)进行定量分析。桃叶珊瑚苷、戟叶马鞭草苷、马鞭草苷、龙胆苦苷在正离子电离模式下定量分析离子对分别为m/z 167.2→m/z 149.1、m/z 243.2→m/z 225.2、m/z 227.2→m/z195.2、m/z 195.2→m/z 177.2。毛蕊花糖苷在负离子电离模式下定量分析离子对为m/z461.3→m/z 161.0。结果桃叶珊瑚苷、戟叶马鞭草苷、马鞭草苷、龙胆苦苷、毛蕊花糖苷五种组分在120~1 200,10 000~100 000,5 000~50 000,1.2~12,510~5 100μg/L的浓度范围内与峰面积均呈良好的线性关系,平均回收率分别为101.2%,98.63%,99.45%,101.6%,100.3%。结论本法操作简便、准确、具有良好的重现性,为马鞭草药材的合理应用及质量控制奠定了基础。     

HPLC测定怀地黄中桃叶珊瑚苷的含量

目的测定不同品种、不同产地怀地黄中桃叶珊瑚苷的含量,初步对怀地黄种质资源进行评价。方法采用HPLC,Dikma Diamonsil C18色谱柱(4.6 mm×250 mm,0.5μm),流动相为甲醇-水(5.5∶94.5),检测波长为206 nm。结果桃叶珊瑚苷在0.053 5~1.284μg间线性关系良好,回归方程为Y=539 696X+3 581.6(r=0.999 9),回收率分别为99.75%(鲜地黄)和99.61%(生地黄),RSD分别为2.86%和2.15%(n=6);不同品种、不同产地怀地黄中桃叶珊瑚苷含量不同。结论本法操作简单、灵敏、稳定,适用于怀地黄中桃叶珊瑚苷的含量测定。     

基于指纹图谱和化学模式识别评价杜仲叶质量

目的 建立杜仲Eucommia ulmoides叶高效液相色谱(HPLC)指纹图谱,结合化学模式识别评价其质量。方法 采用Ultimate XB-C18色谱柱(250 mm×4.6 mm,5μm),以乙腈-0.1%磷酸溶液为流动相梯度洗脱,体积流量1.0 mL/min,柱温30℃,进样量20μL,波长为207、240、277、350 nm,检测杜仲叶中12种活性成分含量并建立指纹图谱,应用SPSS 25.0和SIMCA 14.1软件进行聚类分析(clustering analysis,CA)、主成分分析(principal component analysis,PCA)和偏最小二乘法-判别分析(partial least squares discrimination analysis,PLS-DA),对不同产地杜仲叶进行质量评价。结果 不同产地杜仲叶桃叶珊瑚苷、京尼平苷酸、原儿茶素、绿原酸、儿茶素、车叶草苷、咖啡酸、京尼平苷、松脂醇二葡萄糖苷、芦丁、异槲皮素、紫云英苷差异显著,质量分数范围分别为0.053%～4.563%、0.078%～0.977%、0.018%～0.084%、2.63...     

HPLC法测定鲜地黄中梓醇和桃叶珊瑚苷

目的 建立HPLC法测定鲜地黄中梓醇及桃叶珊瑚苷的方法,对鲜地黄进行质量控制。方法 采用HPLC法对鲜地黄中梓醇及桃叶珊瑚苷进行定量测定。结果 线性回归方程分别为梓醇Y＝43 800 X＋2 849.2,r＝0.999 9;桃叶珊瑚苷Y＝76 163 X＋1 676.1,r＝0.999 9;梓醇和桃叶珊瑚苷线性范围分别为0.19～2.375 μg、0.152～1.90 μg,回收率分别为99.00%和99.38%。结论 该方法准确、重现性好,可以作为鲜地黄的质量控制方法。     

Protective effects of aucubin on H₂O₂-induced apoptosis in PC12 cells

The present study investigated the neuroprotective effects of aucubin on hydrogen peroxide (H₂O₂)‐induced apoptosis in PC12 cells. Exposure of PC12 cells to 0.25 mm H₂O₂ induced a leakage of lactate dehydrogenase and decreased cell viability, as shown by 3‐(4, 5‐dimethylthiazol‐2‐yl)‐2, 5‐diphenyltetrazolium bromide assay. In a dose over 0.1 mm , aucubin increased PC12 cellular viability and markedly attenuated H₂O₂‐induced apoptotic cell death. Quantitation of apoptosis by flow cytometry indicated that aucubin inhibited H₂O₂‐induced apoptosis in PC12 cells. Nuclear damage was alleviated by aucubin, as shown by Hoechst staining. In addition, the levels of malondialdehyde were reduced and the activity of superoxide dismutase, catalase and glutathione peroxidase was augmented in these cells. These results indicated that aucubin inhibited H₂O₂‐induced apoptosis in PC12 cells through regulation of the endogenous oxidant–antioxidant balance. Our results suggest that aucubin is a potential protective agent for the treatment of oxidative‐stress‐induced neurodegenerative disease. Copyright © 2011 John Wiley & Sons, Ltd.     

不同产地杜仲雄花指纹图谱的建立及其化学模式识别研究

目的 建立4个产地21批杜仲Eucommia ulmoides雄花的高效液相色谱(HPLC)指纹图谱,为杜仲雄花的质量控制提供参考。方法 采用HPLC法,色谱柱为Agilent HC-C18色谱柱(250mm×4.6mm,5.0μm);流动相为乙腈-0.1%甲酸水溶液;梯度洗脱;体积流量0.8mL/min;柱温35℃;程序变化波长;进样量20μL。通过相似度评价、偏最小二乘法分析(partial least squares discriminant analysis,PLS-DA)、系统聚类分析(hierarchical cluster analysis,HCA)以及气候因子相关性分析等对杜仲雄花指纹图谱进行分析评价。结果 建立4个产地杜仲雄花HPLC指纹图谱,共标定15个共有峰,指认出其中6个色谱峰,分别为桃叶珊瑚苷、京尼平苷酸、绿原酸、京尼平苷、芦丁、槲皮素;21批杜仲雄花指纹图谱与对照图谱的相似度为0.911~0.989;PLS-DA分析表明不同产地杜仲雄花样品具有一定差异,根据变量投影重要性(variable importance in the projection,VIP)筛选出7个差异性成分;差异性成分峰面积结合气候因子的相关性分析结果<显示9号峰峰面积与3、4月平均最低气温呈显著正相关,15号峰槲皮素峰面积与3月平均降水天数呈显著负相关,其余差异性成分与气候因子未呈现出明显的相关性。结论 该方法可应用于杜仲雄花的质量评价,为杜仲雄花质量控制与资源开发提供参考。     

桃叶珊瑚苷通过激活ERβ途径抑制心肌细胞凋亡

目的探讨桃叶珊瑚苷(aucubin,AU)抑制心肌细胞凋亡发挥抗心肌缺血损伤从而改善急性心肌梗死后心功能的作用与可能机制。方法采用冠状动脉左前降支结扎法制作小鼠急性心肌缺血(myocardial infarction,MI)模型。采用超声心动、Masson染色检测小鼠心功能及梗死面积。建立肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)/放线菌酮(cycloheximide,CHX)诱导的大鼠H9c2心肌细胞损伤模型。采用IncuCyte活细胞成像、TUNEL、Western blot等方法探讨了AU的抗心肌细胞损伤作用及其作用机制。结果研究结果表明,AU可以改善心肌梗死小鼠的心功能,降低梗死面积,减少由TNF-α/CHX诱导的心肌细胞凋亡,降低caspase-3蛋白质表达,上调Bcl2/Bax比值;同时AU可使ERβ(estrogen receptorβ,ERβ)表达升高,且雌激素受体β抑制剂可阻断AU的抗凋亡作用。结论该研究证实,AU可以通过抑制心肌细胞凋亡发挥抗心肌缺血损伤从而改善急性心肌梗死后心功能的作用,其部分作用机制与ERβ通路的激活有关。     

Lipid Nanoparticles Loaded with Selected Iridoid Glycosides as Effective Components of Hydrogel Formulations

One possibility of improving active ingredient penetration into deeper skin layers to enhance the cosmetic product effectiveness, is the application of lipid nanoparticles. The aim of the study presented in this paper was to evaluate the potential of hydrogel formulations enriched with iridoid glycosides-loaded lipid nanoparticles. Lipid nanocarriers were produced using an emulsification-ultrasonication method based on multiple emulsions. The encapsulation efficiency was determined at the level of 89% and 77% for aucubin and catalpol, respectively. The next stage was the incorporation of the obtained dispersions of lipid nanoparticles into hydrogel formulations, followed by determination of their physicochemical properties, shelf-life stability, and application properties (in vivo tests). The introduction of lipid nanoparticles increased the stabilization of the consistency of the obtained hydrogel formulations, and was confirmed by viscosity measurements. No effect of lipid nanoparticle incorporation on shelf-life stability of the hydrogels was detected. In vivo studies showed improvements in moisture content of the epidermis, transepidermal water loss, skin topography, and macrorelief parameters. In particular, a synergistic effect of the active ingredients and lipid nanoparticles on the anti-wrinkle effect, moisturizing effect, and regeneration of the protective barrier of the stratum corneum was evidenced. The attractiveness of aucubin and catalpol as cosmetic raw materials in hydrogel formulations was evidenced, especially when the iridoid glycosides were applied in the form of lipid nanoparticles.     

熟地黄的高效液相色谱／电喷雾电离-质谱分析

目的:建立熟地黄中化学成分的高效液相色谱-电喷雾-质谱分析方法。方法:液相色谱条件为:Zorbax SB-C_(18)色谱柱(250 mm×4.6 mm,5μm);以乙腈-水二元高压梯度洗脱;流速:1 mL·min~(-1);二极管阵列检测器,设定波长:210 nm。质谱条件为:Agilent 离子阱质谱仪;ESI 离子源;负离子检出模式。结果:在负离子检出模式下,熟地黄的总离子流色谱图特征性很强,通过质谱中的主要碎片对梓醇、二氢梓醇、益母草苷、桃叶珊瑚苷以及地黄苷 A、B、C、D 共8种主要成分进行了结构解析。结论:在质谱负离子检出模式下,多数化学成分响应较好,又为熟地黄药材质量控制提供了一种方法。