Beltrame Flávio Luís, Filho Edson Rodrigues, Barros Fabio A Proença, Cortez Diógenes Aparíciogarcia, Cass Quezia Bezerra
Departamento de Química, Universidade Federal de Sao Carlos, SP, Brazil.
J Chromatogr A. 2006 Jun 30;1119(1-2):257-63. doi: 10.1016/j.chroma.2005.10.050. Epub 2005 Dec 19.
The hydroalcoholic extract, prepared from authentic chopped barks of Trichilia catigua, was evaluated by high-performance liquid chromatography using a diode array detector (200-400 mn). The crude extract was purified by rotation locular counter-current chromatography and the chloroform fraction obtained was clean-up by solid-phase extraction. With the aim of getting preliminary structure information on-line, the methanol fraction thus obtained was analyzed by gradient elution using the diode array detector coupled to a mass spectrometer. The presence of flavalignan in this extract was inferred by the chromatographic band, in the total ion current trace, that had an M-H = 451. With this information, cinchonain Ib was isolated as a pure compound from the crude hydroalcoholic extract using a solid-phase extraction procedure for the sample clean-up followed by a semi-preparative separation using the reverse mode of elution. The isolated compound, after complete characterization, was used as an external standard for the development and validation of a method for the analysis of this compound in herbal medicines using the ultraviolet as the detector. The validated method has been successfully applied for quantification of cinchonain Ib in commercialized herbal medicines sold as Catuaba in Brazil and also in standard chopped barks of T. catigua.
从正宗的卡蒂瓜楝（Trichilia catigua）切碎树皮制备的水醇提取物，采用配备二极管阵列检测器（200 - 400纳米）的高效液相色谱法进行评估。粗提取物通过旋转液滴逆流色谱法进行纯化，所得氯仿馏分通过固相萃取进行净化。为了在线获取初步结构信息，使用与质谱仪联用的二极管阵列检测器通过梯度洗脱对由此获得的甲醇馏分进行分析。通过总离子流色谱图中的色谱峰推断该提取物中存在黄酮木脂素，其M - H = 451。基于此信息，使用固相萃取程序对样品进行净化，然后采用反相洗脱的半制备分离方法，从粗水醇提取物中分离出纯化合物辛可宁因Ib。经完全表征后，分离出的化合物用作外标，用于开发和验证一种以紫外为检测器的草药中该化合物分析方法。该验证方法已成功应用于定量巴西作为卡图巴出售的商业化草药以及卡蒂瓜楝标准切碎树皮中的辛可宁因Ib。
