Department of Pharmacology, College of Pharmacy, Fujian Medical University, Fuzhou 350004, Fujian, China.
J Chromatogr A. 2011 Jun 10;1218(23):3695-8. doi: 10.1016/j.chroma.2011.04.025. Epub 2011 Apr 16.
Alkaloids in Gelsemium elegans possess a variety of therapeutic properties, including tumor suppression, analgesic and anti-inflammatory effects. In China, G. elegans has been used for centuries to treat a variety of medical conditions, including chronic pain and skin ulcer. Methods currently used to separate the active components of G. elegans are time-consuming and have low recovery. In the present study, we used pH-zone-refining counter-current chromatography to separate major alkaloids from a crude extract of G. elegans. The two-phase solvent system was methyl tert-butyl ether (MtBE)/acetonitrile/water (3:1.5:4, v/v). Triethylamine (20 mM) was added to the upper organic stationary phase as a retainer. Hydrochloric acid (10 mM) was added to the lower aqueous phase as an eluter. From 1.5 g of crude extract, we obtained 312 mg gelsemine, 420 mg koumine and 195 mg gelsevirine, with purities at 94.8%, 95.9% and 96.7%, respectively, which were determined by HPLC at 256 nm. The chemical identity of the isolated compounds was verified by electrospray ionization-mass spectrometry (ESI-MS), ¹H NMR and ¹³C NMR. These results demonstrated that pH-zone-refining counter-current chromatography is an effective method to separate and purify major alkaloids from G. elegans.
钩吻中的生物碱具有多种治疗特性，包括抑制肿瘤、镇痛和抗炎作用。在中国，钩吻已被使用了几个世纪，用于治疗多种疾病，包括慢性疼痛和皮肤溃疡。目前用于分离钩吻中活性成分的方法耗时且回收率低。在本研究中，我们使用 pH 区带精制逆流色谱法从钩吻粗提取物中分离主要生物碱。两相溶剂系统为甲基叔丁基醚（MtBE）/乙腈/水（3:1.5:4，v/v）。在上相有机固定相中加入三乙胺（20 mM）作为保留剂。在下相水相中加入盐酸（10 mM）作为洗脱剂。从 1.5 g 粗提取物中，我们获得了 312 mg 钩吻碱、420 mg 钩吻明碱和 195 mg 钩吻乙素，纯度分别为 94.8%、95.9%和 96.7%，这是通过在 256 nm 处的 HPLC 确定的。通过电喷雾电离质谱（ESI-MS）、1H NMR 和 13C NMR 验证了分离化合物的化学性质。这些结果表明，pH 区带精制逆流色谱法是一种从钩吻中分离和纯化主要生物碱的有效方法。
