Preparation of Neochlorogenic Acid Reference Substances by Medium-low-pressure Preparative Chromatography
Ni Fuyong;Song Yaling;Liu Lu;Zhao Yiwu;Hong Kui;Huang Wenzhe;Wang Zhenzhong;Xiao Wei;Jiangsu Kanion Pharmaceutical Co., Ltd./ State Key Laboratory of New-tech for Chinese Medicine Pharmaceutical Process;
This study was aimed to establish a separation method for neochlorogenic acid reference substances from Lonicera japonica. Refined neochlorogenic acid in L. japonica water extract was separated and concentrated by HPD200 A macroporous resin, which was isolated and purified by medium-low-pressure preparative chromatography and determined by HPLC. The structure was identified by various spectroscopic data including ESI-MS, 1H-NMR and 13C-NMR. The results showed that the optimal purification technology conditions were as follows: washed with 5BV of water, collected elution, concentration, drying; neochlorogenic acid crude products were eluted with acetonitrile-0.5% formic acid solution(10:90) with the flow rate of 20 m L·min-1; and the detection wavelength was 326 nm. The contents of the prepared neochlorogenic acid reached to 98.86% and the yield was 89.1%. It was concluded that the method was effective for the preparation of neochlorogenic acid with high purity. It can be used to prepare the reference substances for quantitative analysis and content determination of Chinese materia medica.