Qiu Ying-Kun, Chen Fang-Fang, Zhang Ling-Ling, Yan Xia, Chen Lin, Fang Mei-Juan, Wu Zhen
School of Pharmaceutical Sciences and the Key Laboratory for Chemical Biology of Fujian Province, Xiamen University, South Xiang-An Road, Xiamen, 361100, China.
School of Pharmaceutical Sciences and the Key Laboratory for Chemical Biology of Fujian Province, Xiamen University, South Xiang-An Road, Xiamen, 361100, China.
Anal Chim Acta. 2014 Apr 11;820:176-86. doi: 10.1016/j.aca.2014.02.023. Epub 2014 Feb 24.
An on-line comprehensive two-dimensional preparative liquid chromatography system was developed for preparative separation of minor amount components from complicated natural products. Medium-pressure liquid chromatograph (MPLC) was applied as the first dimension and preparative HPLC as the second one, in conjunction with trapping column and makeup pump. The performance of the trapping column was evaluated, in terms of column size, dilution ratio and diameter-height ratio, as well as system pressure from the view of medium pressure liquid chromatograph. Satisfactory trapping efficiency can be achieved using a commercially available 15 mm × 30 mm i.d. ODS pre-column. The instrument operation and the performance of this MPLC×preparative HPLC system were illustrated by gram-scale isolation of crude macro-porous resin enriched water extract of Rheum hotaoense. Automated multi-step preparative separation of 25 compounds, whose structures were identified by MS, (1)H NMR and even by less-sensitive (13)C NMR, could be achieved in a short period of time using this system, exhibiting great advantages in analytical efficiency and sample treatment capacity compared with conventional methods.
开发了一种在线二维制备液相色谱系统，用于从复杂天然产物中制备分离微量成分。以中压液相色谱仪（MPLC）作为第一维，制备型高效液相色谱作为第二维，并结合捕集柱和补充泵。从制备型高效液相色谱的角度，对捕集柱的性能进行了评估，包括柱尺寸、稀释比和径高比以及系统压力。使用市售的内径为15 mm×30 mm的ODS预柱可获得令人满意的捕集效率。通过对河套大黄粗孔树脂富集水提取物进行克级分离，展示了该MPLC×制备型HPLC系统的仪器操作和性能。使用该系统可在短时间内实现25种化合物的自动多步制备分离，这些化合物的结构通过质谱、（1）H核磁共振甚至不太灵敏（13）C核磁共振进行了鉴定，与传统方法相比，在分析效率和样品处理能力方面具有很大优势。
