Isolation of mono-caffeoylquinic acids from tobacco waste using continuous resin-based pre-separation and preparative HPLC.
Three isomers of mono-caffeoylquinic acid, specifically, 3-O-caffeoylquinic acid, 4-O-caffeoylquinic acid and 5-O-caffeoylquinic acid, were successfully isolated from a crude extract of tobacco (Nicotiana tobaccum L.) wastes using continuous resin-based pre-separation and preparative high-performance liquid chromatography (HPLC). The extract of tobacco wastes was continuously pre-separated by resin-based columns packed with D101 and XAD-4, yielding total mono-caffeoylquinic acids with a purity of 67.71% and a recovery rate of 90.06%. Variables affecting resolution and productivity of three mono-caffeoylquinic acid isomers in preparative HPLC (i.e. mobile-phase composition, pH, flow rate and loading amount) were studied. The optimum chromatographic conditions were determined to be a mobile phase consisting of 15% (v/v) methanol and aqueous acetic acid with a pH of 4.5, a flow rate of 4.0 mL/min, a loading amount of 4 mL and a detection wavelength of 360 nm. From 300 mg of loading sample, 56.3 mg of 3-O-caffeoylquinic acid, 92.8 mg of 5-O-caffeoylquinic acid and 73.1 mg of 4-O-caffeoylquinic acid were obtained in a single run, each with a purity of over 98% by HPLC. The structures of the isolated compounds were elucidated by ESI-MS, (1) H-NMR and (13) C-NMR spectral data.
School of Biology and Chemical Engineering, Jiangsu University of Science and Technology, Zhenjiang, P R China. jimwang_js@hotmail.com
Jun
J
Wang
Dingqiang
D
Lu
Yao
Y
Liang
Hui
H
Zhao
Min
M
Luo
Xiuquan
X
Ling
Pingkai
P
Ouyang
D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D007220:Industrial Waste / Q000032:analysis*; D010936:Plant Extracts / Q000032:analysis / Q000302:isolation & purification*; D011801:Quinic Acid / Q000031:analogs & derivatives* / Q000032:analysis / Q000302:isolation & purification; D012117:Resins, Synthetic / Q000138:chemical synthesis; D014026:Nicotiana / Q000737:chemistry*
D007220:Industrial Waste; D010936:Plant Extracts; D012117:Resins, Synthetic; C472707:caffeoylquinic acid; D011801:Quinic Acid