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-H 2 O (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-H 2 O-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-H 2 O (4:1:0.5:5, v/v). The structures of the eight pure compounds were confirmed by electrospray ionization-mass spectrometry (ESI-MS), 1 H-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.

Preparative isolation and purification of 12 main antioxidants from the roots of Polygonum multiflorum Thunb. using high‐speed countercurrent chromatography and preparative HPLC guided by 1,1′‐diphenyl‐2‐picrylhydrazyl‐HPLC

“…In other words, 12 target compounds could not be separated using a single solvent system. To overcome this problem and simultaneously separate compounds with largely different K values, stepwise HSCCC elution mode was proposed [27].…”

Separation of α‐glucosidase inhibitors from Potentilla kleiniana Wight et Arn using solvent and flow‐rate gradient high‐speed counter‐current chromatography target‐guided by ultrafiltration HPLC‐MS screening

“…Thus, HSCCC has been widely used for the separation and purification of active components from natural products, albeit, it is a big challenge to separate all target components directly from crude extract in a single run. Therefore, the stepwise elution mode was introduced into HSCCC separation to address this problem . The common gradient modes used in counter‐current chromatography (CCC) include temperature gradients, stepwise flow‐rate gradients, stepwise solvent gradients, pH‐gradients, and salting‐out gradients.…”

“…Therefore, the stepwise elution mode was introduced into HSCCC separation to address this problem. [18][19][20] The common gradient modes used in counter-current chromatography (CCC) include temperature gradients, stepwise flow-rate gradients, 21,22 stepwise solvent gradients, 20,23 pH-gradients, and salting-out gradients. However, few articles related to HSCCC separation of natural products reported the combination of these gradient modes.…”

Five New Phenolic Compounds with Antioxidant Activities from the Medicinal Insect Blaps rynchopetera

“…Our previous studies have revealed that dipeptides and phenolics—which exhibited various pharmacological activities—are the main active ingredients of this drug [ 5 ]. As part of our continuous work on the systematic study of this insect, the chemical constituents of B. rynchopetera was investigated to afford five new phenolic compounds rynchopeterines A–E ( 1 – 5 ) ( Figure 1 ), along with thirteen known phenolics, procatechuic acid ( 6 ) [ 6 ], 3,4-dihydroxyphenylacetic acid ( 7 ) [ 7 ], 3,4-dihydroxybenzaldehyde ( 8 ) [ 8 ], 3,4-dihydroxyphenylacetaldehyde ( 9 ) [ 9 ], methyl 3,4-dihydroxybenzoate ( 10 ) [ 10 ], 3,4-dihydroxyphenylacetic acid methyl ester ( 11 ) [ 11 ], 3,4-dihydroxyphenylacetic acid ethyl ester ( 12 ) [ 7 ], 2-(3,4-dihydroxyphenyl)ethanol ( 13 ) [ 12 ], 3,4-dihydroxyphenethylacetate ( 14 ) [ 13 ], blapsins A ( 15 ) and B ( 16 ) [ 14 ], 1-(3,4-dimethoxyphenyl)-4-hydroxypentan-1-one ( 17 ) [ 15 ], and 3,3′,4,4′-tetrahydroxybiphenyl ( 18 ) [ 16 ] (see Supplementary ). Herein, the isolation, structural elucidation, and bioactivity of the new compounds are described.…”