高效液相色谱法测定益气口服液中黄芩甙含量

用ＨＰＬＣ法测定了益气口服液中黄芩甙的含量。回归方程Ｙ＝２００１５１ｘ＋３７０１，相关系数γ＝０．９９９５，平均回收率９６．３％（ｎ＝５），ＲＳＤ＝１．８５％。     

云南不同产地灯盏细辛药材中灯盏乙素的HPLC测定

目的：以灯盏乙素的含量为指标来比较云南不同产地灯盏细辛药材的质量差异。方法：灯盏细辛药材经50％乙醇提取，用高效液相色谱方法（C18柱，用甲醇-乙腈-0．5％甲酸为流动相梯度洗脱）测定灯盏乙素的含量。结果：云南不同产地灯盏细辛中灯盏乙素的含量在0．72％～2．95％之间，平均含量1．40％．结论：云南不同产地灯盏细辛药材质量基本一致，以弥勒县等传统产区的药材灯盏乙素含量相对较高。     

6种中药单体对人孕烷X受体介导细胞色素酶P450 3A4转录的调节作用

目的研究6种中药单体野黄芩苷、丹皮酚、芍药苷、甘草苷、黄芪甲苷以及黄芩素是否可以通过诱导孕烷X受体（PXR）介导细胞色素酶P4503A4（CYP3A4）转录表达。方法采用脂质体瞬时转染的方法,将人PXR表达质粒、CYP3A4报告质粒以及内参质粒转入LS174T细胞中,建立报告基因体系,与不同浓度及不同给药时间的中药单体共同孵育,检测细胞中荧光素酶的活性。结果黄芩素（20μmol/L、40μmol/L和80μmol/L）分别诱导PXR介导CYP3A4表达（1.49±0.23）倍（P〈0.05）、（2.45±0.56）倍（P〈0.01）和（5.27±0.40）倍（P〈0.01）,野黄芩苷、丹皮酚、芍药苷、甘草苷和黄芪甲苷在实验的浓度范围内不能明显诱导LS174T细胞中PXR介导CYP3A4表达。在不同给药时间实验中,20μmol/L、40μmol/L和80μmol/L的黄芩素在12～48h能诱导PXR介导CYP3A4表达,诱导能力随时间的延长而呈增强的趋势,各浓度的最大诱导倍数均在48h出现。结论黄芩素在LS174T细胞系中可以通过诱导PXR介导CYP3A4转录表达,而野黄芩苷、丹皮酚、芍药苷、甘草苷或黄芪甲苷无此作用。     

野黄芩苷抗血小板活化效应的评价

目的评价野黄芩苷对血小板的聚集与活化的影响。方法在人洗涤血小板中进行血小板聚集实验,分别用胶原(1.00μg/ml)、U46619(0.30μM)、ADP(10.00μM)、凝血酶(0.04U/ml)作为诱导剂,对比在0.04mg/ml、0.10mg/ml、0.20mg/ml三种不同预孵浓度状态下野黄芩苷对血小板聚集的影响。并进一步研究三种不同浓度的野黄芩苷对血栓素类似物(U46619)引起的血小板纤维蛋白原结合的影响。结果胶原、U46619、ADP、凝血酶均可明显促进血小板聚集,而血小板预孵育野黄芩苷可以抑制诱导剂引起的聚集,并且抑制作用呈现浓度依赖关系。血小板预孵育野黄芩苷可以减少U46619引起的血小板纤维蛋白结合,并且抑制作用呈现浓度依赖关系。结论野黄芩苷可以抑制血小板的聚集和活化,很可能是通过阻断血栓素A2诱导血小板聚集的某一个中间环节产生作用。     

Designing novel pH-sensitive non-phospholipid vesicle: Characterization and cell interaction

In an effort to enhance the oral bioavailability of scutellarin, ethyl, benzyl and N,N-diethylglycolamide ester of scutellarin were synthesized. The hydrolysis of the prodrugs follows first-order kinetics in aqueous solution, and produced a V-shaped pH profile. The N,N-diethylglycolamide ester is highly susceptible to enzymatic hydrolysis in human plasma (t_1/2 ≈ 7 min) with a high stability in aqueous solution (t_1/2 ≈ 16 day, pH 4.2). Compared with the solubility of scutellarin, the solubility of glycolamide ester was about ten times in pH 4.0 buffer, and about thirty five times in water. Its apparent partition coefficient increased significantly from −2.56 to 1.48. Glycolamide ester of scutellarin was chosen to investigate the intestinal metabolism and in vivo bioavailability. Degradation studies in the intestinal tract content and homogenates indicated intestinal metabolism before absorption was a crucial obstacle for the prodrug. N,N-Diethylglycolamide ester can be protected from the degradation in the intestinal lumen by an emulsion. A significant increase in the plasma AUC and C_max of the prodrug emulsion was observed in rats, compared with that of the scutellarin–cyclodextrin complex (P < 0.01). The emulsion of N,N-diethylglycolamide ester produces a 1.58-fold enhancement in apparent bioavailability and 1.4-fold increase in the absolute bioavailability compared to the scutallarin–cyclodextrin complex.     

HPLC法测定灯盏生脉胶囊中野黄芩苷的含量

目的建立HPLC的方法测定灯盏生脉胶囊中野黄芩苷的含量。方法选用Agilentzorbax SB-C18色谱柱（250mm×4.6mm,5μm）;柱温：35℃;测定波长：333nm;流动相：0.1%磷酸水溶液（C）-乙腈（D）,梯度洗脱;流量：1mL·min-1为测定条件。结果野黄芩苷浓度在6～60μg·mL-1范围内与峰面积呈良好线性关系,r=0.9996。野黄芩苷的平均加样回收率为100.5%,RSD=2.89%。结论该方法可靠性高,准确性和重现性好,可作为灯盏生脉胶囊的质量控制标准。     

灯盏花素片溶出度测定方法的考察

目的：建立灯盏花素片溶出度测定的方法。方法：以磷酸盐缓冲液（pH6．8）为溶出介质,按《中国药典}2010年版二部附录XC第二法操作,用紫外分光光度法检测,计算灯盏花素片的溶出度。结果：灯盏花素在5．30-12．38μg·ml^-1范围内浓度和吸光度成良好线性,r=0．9999。本法平均回收率为97．25％,RSD为0．94％。溶出度受转速影响小,受溶剂影响大。结论：本方法简单、准确、重复性好,可用于灯盏花素片的质量控制。     

柱层析法纯化灯盏乙素的研究

目的优选灯盏乙素的柱层析纯化条件。方法采用高效液相色谱法检测纯度,考察了不同固定相和流动相对灯盏乙素分离效果的影响。结果灯盏乙素的最佳纯化工艺条件为:固定相200-300目的硅胶柱,流动相V(CH2Cl2)∶V(CH3OH)=5∶1,常压洗脱。结论该方法操作简单,分离效果好,使用成本低。     

Study on metabolism of scutellarin in rats by HPLC-MS and HPLC-NMR

Scutellarin is the major active constituent of Scutellaria barbata D. The metabolism of scutellarin has been investigated in rats. The solid-phase extraction and HPLC-DAD methods were established to separate and analyse metabolites. Five metabolites (M1–M5) were identified by enzymatic hydrolysis, HPLC-DAD, HPLC-MS and HPLC-NMR. M1 and M3 were conjugates of scutellarin with two sulfate groups, which have not been reported in natural plants. M2 was scutellarin; M4 was 6-methyl-scutellarin; and M5 was 6-methyl-scutellarein. The metabolic pathway was proposed.     

灯盏花素缓释片在Beagle犬体内相对生物利用度的研究

目的　探讨灯盏花素缓释片在动物体内的药物动力学规律 ,并进行相对生物利用度研究。方法　采用双周期交叉试验设计 ,对Beagle犬单剂量顿服灯盏花素缓释片和参比制剂的动力学参数进行比较。结果　采用尿药法进行数据处理 ,参比制剂与供试制剂的尿药总排泄量 (Xu)分别为 375 92 5± 4 7 2 3μg和 394 5 92± 32 84 μg ;生物半衰期 (t1/2 )分别为 7 197± 0 5 91h和 12 6 2± 1 2 0 7h。结论　以吸收总量为指标 ,灯盏花素缓释片与参比制剂相比是生物等效制剂 ,并具有缓释特征。