循环制备液相色谱分离芳香新塔花中的化学成分

Isolation of chemical constituents from Ziziphora clinopodioides Lam. with recycling preparative high performance liquid chromatography

建立了交替循环和直接循环液相色谱相结合的方法用于制备芳香新塔花中的化学成分。芳香新塔花样品经溶剂提取、柱色谱和中压制备色谱初步分离后得到芳香新塔花的不同馏分。以甲醇-水为流动相,利用双柱交替循环法对组分进行分离,同时,流动相经恒流泵循环输入色谱柱。以馏分Ⅰ和馏分Ⅱ为例,在混合循环模式下分离得到5个化合物。通过核磁共振对其进行鉴定,确定分别为乔松素-7-O-芸香糖苷、白杨素-7-O-芸香糖苷、金合欢素-7-O-芸香糖苷、云杉素和原儿茶酸。实验结果表明,该制备方法分离效率高,节省流动相,是分离天然产物的有效手段。

The combination of alternate recycling and direct recycling preparative liquid chro-matography method was developed for the isolation of chemical constituents from Ziziphora clinopodioides Lam. The crude extract was obtained from Ziziphora clinopodioides Lam. by solvent extraction,column chromatography and reversed-phase（ RP ） flash chromatography. All the separations were performed with methanol and water as mobile phases and the devel-oped recycling preparative method was used with twin RP columns switched by a two-position ten-way valve for the separation. The mobile phase was recycled in close loop with a two-posi-tion six-way valve. The fraction Ⅰ and fraction Ⅱ from reversed-phase flash chromatography were selected for the demonstration of separation power of the proposed protocol,and five compounds were obtained from Ziziphora clinopodioides Lam. The isolated five compounds were identified as pinocembrin-7-O-rutinoside,pinocembrin-7-O-rutinoside,acacetin-7-O-ruti-noside,picein and protocatechuic acid with nuclear magnetic resonance（ NMR）. The experi-mental results showed that the developed preparation method exhibited higher separation effi-ciency with less mobile phase used than the reported methods,and could be expected as an effective method for the separation of complex natural products,especially the compounds with similar structures.