An Efficient Method for the Preparative Isolation and Purification of Flavonoid Glycosides and Caffeoylquinic Acid Derivatives from Leaves of Lonicera japonica Thunb. Using High Speed Counter-Current Chromatography (HSCCC) and Prep-HPLC Guided by DPPH-HPLC Experiments.
In this work, the n-butanol extract from leaves of Lonicera japonica Thunb. (L. japonica) was reacted with DPPH and subjected to a HPLC analysis for the guided screening antioxidants (DPPH-HPLC experiments). Then, nine antioxidants, including flavonoid glycosides and caffeoylquinic acid derivatives, were isolated and purified from leaves of L. japonica using high speed counter-current chromatography (HSCCC) and prep-HPLC. The n-butanol extract was firstly isolated by HSCCC using methyl tert-butyl ether/n-butanol/acetonitrile/water (0.5% acetic acid) (2:2:1:5, v/v), yielding five fractions F1, F2 (rhoifolin), F3 (luteoloside), F4 and F5 (collected from the column after the separation). The sub-fractions F1, F4 and F5 were successfully separated by prep-HPLC. Finally, nine compounds, including chlorogenic acid (1), lonicerin (2), rutin (3), rhoifolin (4), luteoloside (5), 3,4-Odicaffeoylquinic acid (6), hyperoside (7), 3,5-O-dicaffeoylquinic acid (8), and 4,5-O-dicaffeoylquinic acid (9) were obtained, respectively, with the purities over 94% as determined by HPLC. The structures were identified by electrospray ionization mass spectrometry (ESI-MS), 1H- and 13C-NMR. Antioxidant activities were tested, and the isolated compounds showed strong antioxidant activities.
Shandong Analysis and Test Center, Shandong Academy of Sciences, 19 Keyuan Street, Jinan 250014, China. wangdaijie@126.com.
Daijie
D
Wang
Beijing Centre for Physical and Chemical Analysis, Beijing 100089, China. maggieduning@163.com.
Ning
N
Du
Shandong Analysis and Test Center, Shandong Academy of Sciences, 19 Keyuan Street, Jinan 250014, China. 18663081657@163.com.
Lei
L
Wen
Shandong Analysis and Test Center, Shandong Academy of Sciences, 19 Keyuan Street, Jinan 250014, China. sdzbzdzh@sina.com.
Heng
H
Zhu
Shandong Analysis and Test Center, Shandong Academy of Sciences, 19 Keyuan Street, Jinan 250014, China. liufeng8109@163.com.
Feng
F
Liu
Shandong Analysis and Test Center, Shandong Academy of Sciences, 19 Keyuan Street, Jinan 250014, China. wxjn1998@126.com.
Xiao
X
Wang
College of Food Science and Engineering, Shandong Agricultural University, 61 Daizong Street, Taian 271018, China. djh@sdau.edu.cn.
Jinhua
J
Du
Shandong Yate Eco-Tech Co. Ltd., Linyi 266071, China. yate886@sohu.com.
Shengbo
S
Li
D000975:Antioxidants / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology; D002851:Chromatography, High Pressure Liquid; D003377:Countercurrent Distribution; D005419:Flavonoids / Q000737:chemistry* / Q000302:isolation & purification* / Q000494:pharmacology; D006027:Glycosides / Q000737:chemistry* / Q000302:isolation & purification* / Q000494:pharmacology; D031239:Lonicera / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D015394:Molecular Structure; D010936:Plant Extracts / Q000737:chemistry; D018515:Plant Leaves / Q000737:chemistry*; D011801:Quinic Acid / Q000031:analogs & derivatives* / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology; D012997:Solvents; D021241:Spectrometry, Mass, Electrospray Ionization
D000975:Antioxidants; D005419:Flavonoids; D006027:Glycosides; D010936:Plant Extracts; D012997:Solvents; C472707:caffeoylquinic acid; D011801:Quinic Acid
DPPH-HPLC experiments; Lonicera japonica Thunb.; caffeoylquinic acid derivatives; flavonoid glycosides; high speed counter-current chromatography; prep-HPLC combination