Ye Haoyu, Ignatova Svetlana, Luo Houding, Li Yanfang, Peng Aihua, Chen Lijuan, Sutherland Ian
State key laboratory of Biotherapy, West China Hospital, West China Medical School, Sichuan University, Chengdu 610041, China.
J Chromatogr A. 2008 Dec 12;1213(2):145-53. doi: 10.1016/j.chroma.2008.09.116. Epub 2008 Oct 17.
This paper describes how high-performance counter-current chromatography (HPCCC) was used strategically for the separation of Tripterygium wilfordii Hook. f. Due to the complexity of Chinese herbal medicines, the initial ethanol crude extract was fractionated into seven fractions using medium-pressure liquid chromatography (MPLC). One terpenoid (triptolide) and three alkaloids (peritassine A, wilforgine and wilforine) were further separated from one of the MPLC fractions. This fraction (1.25 g) yielded 8 mg of triptolide and 28 mg of peritassines A after one HPCCC column pass and 30 mg of wilforgine and 120 mg of wilforine after a second column pass with respective purities of 97%, 93.6%, 95.0% and 94.4%, which were determined by high-performance liquid chromatography (HPLC). This was a one-step HPCCC separation, using an n-hexane-ethyl acetate-methanol-water (4:5:4:5, v/v) solvent system, where increases in theoretical plates have been sacrificed in favour of increasing throughput. Structures were identified by electrospray ionization mass spectrometry (ESI-MS), (1)H nuclear magnetic resonance ((1)H NMR) and (13)C nuclear magnetic resonance ((13)C NMR). Comparison of three different modes of eluting compounds retained in the liquid stationary phase: elution extrusion; dual mode and simple pump-out showed that simply pumping out the column contents at high flow gave better resolution and was eight times faster than the other two well-utilised methods. Triptolide and peritassines A were isolated for the first time from Tripterygium wilfordii Hook. f.
本文描述了如何战略性地使用高效逆流色谱法（HPCCC）分离雷公藤（Tripterygium wilfordii Hook. f.）。由于中草药成分复杂，最初的乙醇粗提物采用中压液相色谱法（MPLC）分离成七个馏分。从其中一个MPLC馏分中进一步分离出一种萜类化合物（雷公藤内酯醇）和三种生物碱（雷公藤辛碱A、雷公藤吉碱和雷公藤宁碱）。该馏分（1.25 g）经一次HPCCC柱分离后得到8 mg雷公藤内酯醇和28 mg雷公藤辛碱A，第二次柱分离后得到30 mg雷公藤吉碱和120 mg雷公藤宁碱，其纯度分别为97%、93.6%、95.0%和94.4%，这些纯度通过高效液相色谱法（HPLC）测定。这是一次HPCCC一步分离，使用正己烷 - 乙酸乙酯 - 甲醇 - 水（4:5:4:5，v/v）溶剂系统，在此过程中牺牲了理论塔板数的增加以提高通量。通过电喷雾电离质谱（ESI-MS）、氢核磁共振（¹H NMR）和碳核磁共振（¹³C NMR）鉴定结构。比较了在液体固定相中保留的化合物的三种不同洗脱模式：洗脱挤压；双模式和简单泵出，结果表明在高流速下简单地泵出柱内容物具有更好的分离度，并且比其他两种常用方法快八倍。雷公藤内酯醇和雷公藤辛碱A首次从雷公藤（Tripterygium wilfordii Hook. f.）中分离得到。
