title: Preparative Separation of Phenolic Compounds from Chimonanthus praecox Flowers by High-Speed Counter-Current Chromatography Using a Stepwise Elution Mode.
abstract: High-speed counter-current chromatography (HSCCC) has been successfully used for the separation of eight compounds from Chimonanthus praecox flowers. Firstly, the crude extract of Chimonanthus praecox flowers was dissolved in a two-phase solvent system composed of petroleum ether-ethyl acetate-methanol-H₂O (5:5:3:7, v/v) and divided into two parts: the upper phase (part I) and the lower phase (part II). Then, HSCCC was applied to separate the phenolic acids from part I and part II, respectively. Considering the broad polarity range of target compounds in part I, a stepwise elution mode was established. Two optimal solvent systems of petroleum ether-ethyl acetate-methanol-H₂O-formic acid (FA) (5:5:3:7:0.02, 5:5:4.3:5.7:0.02, v/v) were employed in this separation. Five phenylpropanoids and two flavonoids were successfully separated from 280 mg of part I, including 8.7 mg of 3,4-dihydroxy benzoic acid (a, 95.3% purity), 10.9 mg of protocatechualdehyde (b, 96.8% purity), 11.3 mg of p-coumaric acid (c, 98.9% purity), 12.2 mg of p-hydroxybenzaldehyde (d, 95.9% purity), 24.7 mg of quercetin (e, 97.3% purity), 33.8 mg of kaempferol (f, 96.8% purity), and 24.6 mg of 4-hydroxylcinnamic aldehyde (g, 98.0% purity). From 300 mg of part II, 65.7 mg of rutin (h, 98.2% purity), 7.5 mg of 3,4-dihydroxy benzoic acid (a, 77.4% purity), and 4.7 mg of protocatechualdehyde (b, 81.6% purity) were obtained using the solvent system EtOAc-n-butanol (n-BuOH)-FA-H₂O (4:1:0.5:5, v/v). The structures of the eight pure compounds were confirmed by electrospray ionization-mass spectrometry (ESI-MS), ¹H-NMR and (13)C-NMR. To the best of our knowledge, compounds a-d and f were the first separated and reported from the Chimonanthus praecox flower extract.
authors.affiliation: College of Pharmacy, Shandong University of Traditional Chinese Medicine, 4655 Daxue Street, Jinan 250355, Shandong, China. huaizhili123@sina.com.|Key Laboratory of TCM Quality Control Technology, Shandong Analysis and Test Center, Shandong Academy of Sciences, 19 Keyuan Street, Jinan 250014, Shandong, China. huaizhili123@sina.com.
authors.forename: Huaizhi
authors.initials: H
authors.lastname: Li
authors.affiliation: College of Pharmacy, Shandong University of Traditional Chinese Medicine, 4655 Daxue Street, Jinan 250355, Shandong, China. zyq622003@126.com.
authors.forename: Yongqing
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Key Laboratory of TCM Quality Control Technology, Shandong Analysis and Test Center, Shandong Academy of Sciences, 19 Keyuan Street, Jinan 250014, Shandong, China. 18765878227@126.com.
authors.forename: Qian
authors.initials: Q
authors.lastname: Liu
authors.affiliation: College of Pharmacy, Shandong University of Traditional Chinese Medicine, 4655 Daxue Street, Jinan 250355, Shandong, China. scllbz@163.com.|Key Laboratory of TCM Quality Control Technology, Shandong Analysis and Test Center, Shandong Academy of Sciences, 19 Keyuan Street, Jinan 250014, Shandong, China. scllbz@163.com.
authors.forename: Changlei
authors.initials: C
authors.lastname: Sun
authors.affiliation: College of Pharmacy, Shandong University of Traditional Chinese Medicine, 4655 Daxue Street, Jinan 250355, Shandong, China. wangx@sdas.org.
authors.forename: Jia
authors.initials: J
authors.lastname: Li
authors.affiliation: Kangsen Sanfeng Biological Engineering Technology Co., Jinan 250014, Shandong, China. Pengy@sina.com.
authors.forename: Peng
authors.initials: P
authors.lastname: Yang
authors.affiliation: Key Laboratory of TCM Quality Control Technology, Shandong Analysis and Test Center, Shandong Academy of Sciences, 19 Keyuan Street, Jinan 250014, Shandong, China. Ljytl7172@163.com.
authors.forename: Xiao
authors.initials: X
authors.lastname: Wang
mesh_terms: D031237:Calycanthaceae / Q000737:chemistry*; D002853:Chromatography, Liquid; D003377:Countercurrent Distribution; D035264:Flowers / Q000737:chemistry*; D015394:Molecular Structure; D010636:Phenols / Q000737:chemistry* / Q000302:isolation & purification*; D010936:Plant Extracts / Q000737:chemistry / Q000302:isolation & purification; D066244:Proton Magnetic Resonance Spectroscopy; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D010636:Phenols; D010936:Plant Extracts
keywords: Chimonanthus praecox flower; high-speed counter-current chromatography (HSCCC); phenolic compounds; preparative separation; stepwise elution mode