Central Laboratory, Changchun Normal University, 677 North Changji Road, Changchun 130032, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2010 Nov 15;878(30):3149-55. doi: 10.1016/j.jchromb.2010.09.027. Epub 2010 Oct 19.
High-performance counter-current chromatography (HPCCC) and high performance liquid chromatography coupled with mass spectrometry (HPLC-MS) was efficiently utilized for the separation and identification of the chemical components with a wide range of polarity from the mixed extract of Chinese medicinal herb Apocynum venetum. For HPCCC separation, four sets of solvent systems, n-hexane-ethyl acetate-acetonitrile-water (1.5:3.5:2:4.5, v:v:v:v), ethyl acetate-methanol-water (5:2:5, v:v:v) and n-butanol-methanol-water (5:1:5, v:v:v) were used for the one-step separation by four stages. The HPCCC separation was initiated by filling the column with the lower phase of n-hexane-ethyl acetate-acetonitrile-water (1.5:3.5:2:5, v:v:v:v) as a stationary phase followed by elution with the upper phase of n-hexane-ethyl acetate-acetonitrile-water (1.5:3.5:2:5, v:v:v:v) to separate the hydrophobic compounds (tail to head). Then the mobile phase was switched to the upper phase of ethyl acetate-acetonitrile-water (5:3:7, v:v:v) to eluted the moderate hydrophobic compounds, then the mobile phase was switched to the upper phase of ethyl acetate-methanol-water (5:2:5, v:v:v) to eluted the moderate hydrophilic compounds, and finally the hydrophilic compounds still retained in the column was eluted by the upper phase of n-butanol-methanol-water (5:1:5, v:v:v). A total of 16 named compounds including adhyperforin, hyperforin, amentoflavone, biapigenin, quercetin, avicularin, acetylated isoquercetin, acetylated hyperoside, astragalin, trifolin, isoquercetin, hyperoside, querciturone, rutin, chlorogenic acid and quercetin-3-O-β-D-glucosyl-β-D-glucopyranoside were successfully separated via the four sets of solvent systems in one step operation for 130 min. The compounds separated by HPCCC were identified by comparing with mixed standards data of HPLC-MS as well as NMR data.
高效逆流色谱（HPCCC）和高效液相色谱与质谱联用（HPLC-MS）被有效地用于从中药白屈菜的混合提取物中分离和鉴定具有广泛极性的化学成分。对于 HPCCC 分离，使用了四组溶剂系统，正己烷-乙酸乙酯-乙腈-水（1.5:3.5:2:4.5，v:v:v:v）、乙酸乙酯-甲醇-水（5:2:5，v:v:v）和正丁醇-甲醇-水（5:1:5，v:v:v），通过四个阶段的一步分离。HPCCC 分离是通过用正己烷-乙酸乙酯-乙腈-水（1.5:3.5:2:5，v:v:v:v）的下相作为固定相填充柱开始的，然后用正己烷-乙酸乙酯-乙腈-水（1.5:3.5:2:5，v:v:v:v）的上相洗脱，以分离疏水性化合物（从尾部到头部）。然后将流动相切换到乙酸乙酯-乙腈-水（5:3:7，v:v:v）的上相，洗脱中等疏水性化合物，然后将流动相切换到乙酸乙酯-甲醇-水（5:2:5，v:v:v）的上相，洗脱中等亲水性化合物，最后亲水化合物仍保留在柱中，用正丁醇-甲醇-水（5:1:5，v:v:v）的上相洗脱。通过四组溶剂系统在一步操作中，成功地在 130 分钟内分离出 16 种命名化合物，包括阿魏酸佛司可林、原佛司可林、芹菜素、白杨素、槲皮素、鸡纳皮苷、乙酰化异槲皮苷、乙酰化金丝桃苷、黄芪苷、三福宁、异槲皮苷、金丝桃苷、槲皮酮、芦丁、绿原酸和槲皮素-3-O-β-D-葡萄糖基-β-D-吡喃糖苷。通过与 HPLC-MS 的混合标准数据以及 NMR 数据比较，鉴定了 HPCCC 分离的化合物。
