State Key Laboratory of Chemical Resource Engineering, Beijing University of Chemical Technology, Beijing, China.
J Sep Sci. 2013 Jul;36(13):2210-5. doi: 10.1002/jssc.201300260. Epub 2013 May 27.
Chlorogenic acid and caffeic acid were selected as test samples for separation by the pH-zone-refining countercurrent chromatography (CCC). The separation of these test samples was performed with a two-phase solvent system composed of methyl-tert-butyl-ether/acetonitrile/water at a volume ratio of 4:1:5 v/v/v where trifluoroacetic acid (TFA; 8 mM) was added to the organic stationary phase as a retainer and NH4 OH (10 mM) to the aqueous mobile phase as an eluter. Chlorogenic acid was successfully separated from Flaveria bidentis (L.) Kuntze (F. bidentis) and Lonicerae Flos by pH-zone-refining CCC, a slightly polar two-phase solvent system composed of methyl-tert-butyl-ether/acetonitrile/n-butanol/water at a volume ratio of 4:1:1:5 v/v/v/v was selected where TFA (3 mM) was added to the organic stationary phase as a retainer and NH4 OH (3 mM) to the aqueous mobile phase as an eluter. A 16.2 mg amount of chlorogenic acid with the purity of 92% from 1.4 g of F. bidentis, and 134 mg of chlorogenic acid at the purity of 99% from 1.3 g of crude extract of Lonicerae Flos have been obtained. These results suggest that pH-zone-refining CCC is suitable for the isolation of the chlorogenic acid from the crude extracts of F. bidentis and Lonicerae Flos.
绿原酸和咖啡酸被选为 pH 区带精制逆流色谱 (CCC) 分离的测试样品。这些测试样品的分离是在两相溶剂系统中进行的，该系统由甲基叔丁基醚/乙腈/水组成，体积比为 4:1:5 v/v/v，其中三氟乙酸 (TFA；8 mM) 添加到有机固定相中作为保留剂，NH4 OH（10 mM）添加到水相作为洗脱剂。通过 pH 区带精制 CCC 成功地从 F. bidentis（L.）Kuntze（F. bidentis）和金银花中分离出绿原酸，选择了一个稍微极性的两相溶剂系统，由甲基叔丁基醚/乙腈/正丁醇/水组成，体积比为 4:1:1:5 v/v/v/v，其中 TFA（3 mM）添加到有机固定相中作为保留剂，NH4 OH（3 mM）添加到水相作为洗脱剂。从 1.4 g F. bidentis 中获得了 16.2 mg 纯度为 92%的绿原酸，从 1.3 g 金银花粗提取物中获得了 134 mg 纯度为 99%的绿原酸。这些结果表明，pH 区带精制 CCC 适合从 F. bidentis 和金银花的粗提取物中分离绿原酸。
