秦岭龙胆的化学成分

从秦岭龙胆(Gentiana apiata N.E.Br.)全草的乙醇提取物中分离得到4个化合物,通过波谱分析分别鉴定为异荭草甙(isoorientin),龙胆苦甙(gentiopicroside),asystasioside A和蔗糖(D-sucrose).采用二维NMR分析,确定了asystasioside A的立体结构.以上化合物均为首次从该植物中发现.     

Identification of bio‐active metabolites of gentiopicroside by UPLC/Q‐TOF MS and NMR

Gentiopicroside (GPS), the main bioactive component in Gentiana scabra Bge., has attracted our attention owing to its high bioactivity, especially the treatment of hepatobiliary disorders. The aglycone form of GPS, a typical secoiridoid glycoside, is considered to be more readily absorbed than its parent drug. This study aimed to identify and characterize the metabolites after GPS incubated with β‐glucosidase in buffer solution at 37°C. Samples of biotransformed solution were collected and analyzed by ultraperformance liquid chromatography (UPLC)/quadrupole–time‐of‐flight mass spectrometry (Q‐TOF MS). A total of four metabolites were detected: two were isolated and elucidated by preparative‐HPLC and NMR techniques, and one of those four is reported for the first time. The mass spectral fragmentation pattern and accurate masses of metabolites were established on the basis of UPLC/Q‐TOF MS analysis. Structure elucidation of metabolites was achieved by comparing their fragmentation pattern with that of the parent drug. A fairly possible metabolic pathway of GPS by β‐glucosidase was proposed. The hepatoprotective activities of metabolites M1 and M2 were investigated and the results showed that their hepatoprotective activities were higher than that of parent drug. Our results provided a meaningful basis for discovering lead compounds from biotransformation related to G. scabra Bge. in traditional Chinese medicine. Copyright © 2013 John Wiley & Sons, Ltd.     

固相萃取/超高效液相色谱法测定龙胆泻肝丸中栀子苷、龙胆苦苷与黄芩苷

建立了一种快速测定龙胆泻肝丸中栀子苷、龙胆苦苷和黄芩苷3种有效成分的超高效液相色谱方法。样品经50%甲醇提取,C18固相萃取(SPE)小柱净化后,采用Ultimate XB-C18色谱柱(4.6 mm×50 mm,1.8μm)进行分离,以乙腈-0.1%磷酸溶液为流动相进行梯度洗脱。考察了不同规格色谱柱、流动相梯度比例、进样体积、柱温和流速对分离效果的影响,并对SPE柱流出液的平衡体积进一步考察,在最佳分析条件下进行液相色谱分析,该方法显示了良好的线性关系(r≥0.999 8),其定量下限(LOQ)为0.24～0.44 mg/L,加标回收率为95%～99%。该方法快速、准确,可满足实际检测需要。     

制备型高效液相色谱法分离纯化川西獐牙菜提取物中的龙胆苦苷

龙胆苦苷(GPS)是龙胆类药材及其相关制品质量控制的指标成分。本研究利用制备型高效液相色谱从川西獐牙菜提取物中分离纯化龙胆苦苷对照品。对制备色谱的流动相组成、流速、进样量和检测波长等制备参数进行了优化。采用的色谱柱为C18柱(200 mm×50 mm, 5 μm),流动相为甲醇和0.1%乙酸水溶液(体积比为30:70),流速为75 mL/min,检测波长为254 nm,进样体积为500 μL。在30 min的运行时间内,龙胆苦苷与其他干扰成分得到了很好的分离,产品纯度达到了99%以上。此方法具有快速高效、产品纯度高的特点,可用于制备龙胆苦苷对照品和对龙胆苦苷制品的质量控制。     

高速逆流色谱法从秦艽地上部分制备分离龙胆苦苷

采用高速逆流色谱法分离纯化秦艽地上部分中的龙胆苦苷。溶剂系统为V(氯仿)∶V(甲醇)∶V(水)=4∶4∶2,上相为固定相,下相为流动相,转速为800r/min,流速为2mL/min。所得产物经LC-MS分析为龙胆苦苷,纯度经高效液相色谱分析为94.0%(峰面积归一化法)。在保证分离效果的前提下,增大进样量,摸索制备分离龙胆苦苷的高速逆流色谱条件,为用高速逆流色谱法大量制备龙胆苦苷提供了一个可行方法。