Separation of polyphenols from leaves of Malus hupehensis (Pamp.) Rehder by off-line two-dimensional High Speed Counter-Current Chromatography combined with recycling elution mode

a b s t r a c tAccording to recent studies, the antioxidant and anticancer related properties of European hornbeam (Carpinus betulus L.) leaf extracts are remarkable, making it a possible raw material for medicine, requiring the identification of major active compounds and the assessment of the antioxidant properties of the extracts. In the present work the high-performance liquid chromatographic/multistage mass spectrometric characterization of the antioxidant polyphenolic compounds as well as the evaluation of the seasonal changes in the major antioxidant capacity and related parameters (DPPH, ABTS, FRAP, total phenol content, total flavonoid content and total flavan-3-ol content) have been carried out for the leaves of European hornbeam for the first time. The extracts exhibited the highest antioxidant capacity in August, basing on the FRAP (106.24 ± 3.10 mg AAE/g dw.) and DPPH (IC 50 : 4.63 ± 0.88 g/ml) assays and in May, based on the ABTS (329.78 ± 23.89 mg TE/g dw.) method. From the August extract a total of 171 compounds, including phenolic acids, flavonoid glycosides, tannins, catechins and procyanidins have been characterized and identified. According to the HPLC-PDA peak areas, the most abundant polyphenolic compounds in the August extracts were chlorogenic acid, ellagic acid, ellagitannins, myricetin-, luteolin-, quercetin-and apigenin glycosides. Further research is needed for the elucidation of the compounds which are primarily responsible for the antioxidant properties.

Industrial Crops and Products

Section: Introductionmentioning

confidence: 99%

Antioxidant properties and detailed polyphenol profiling of European hornbeam (Carpinus betulus L.) leaves by multiple antioxidant capacity assays and high-performance liquid chromatography/multistage electrospray mass spectrometry

Hofmann

Nebehaj

Albert

2016

“…According to the base peak 1,3 A – in MS/MS spectra, two flavones, including chrysin ( 8 ) and isoetin‐5’‐ O ‐glucopyranoside ( 25 ), could be tentatively identified by comparing their chromatographic characteristics, absorption spectra, and MS/MS spectra with those of standards. And the structures of five flavonols, avicularin ( 12 ) , kaempferol‐3‐ O ‐β‐ d ‐glycosidase ( 13 ) , quercetin‐3‐ O ‐β‐ d ‐glycosidease ( 19 ), quercetin‐3‐ O ‐β‐ d ‐glycosidease isomer ( 20 ), and kaempferol‐3‐ O ‐rhamnoside ( 24 ), could be preliminarily elucidated based on their t R , UV spectra, and MS/MS spectra with those of standards. The base peaks in MS/MS spectra for them were 1,3 A – and 1,2 A – .…”

Section: Resultsmentioning

confidence: 99%

Rapid screening and identification of antioxidants in the leaves of Malus hupehensis using off‐line two‐dimensional HPLC–UV–MS/MS coupled with a 1,1′‐diphenyl‐2‐picrylhydrazyl assay

Huang

J of Separation Science

et al. 2018

Self Cite

The leaves of Malus hupehensis have a strong antioxidant activity and are commonly consumed as a healthy tea. However, detailed information about its antioxidants is incomplete. Herein, we developed an effective strategy based on combining off-line two-dimensional high-performance liquid chromatography with ultraviolet and tandem mass spectrometry detection with a 1,1'-diphenyl-2-picrylhydrazyl assay to rapidly screen and identify the antioxidants from the leaves of M. hupehensis. In the orthogonal two-dimensional liquid chromatography system, a Venusil HILIC column was used for the first dimension, while a Universil XB-C column was installed in the second dimension. As a result, 32 antioxidants, including ten dihydrochalcones, two flavanones, nine flavonols, four flavones, and seven phenolic acids were tentatively identified, out of which 23 compounds, as far as we know, were isolated and characterized from the leaves of M. hupehensis for the first time. To the best of our knowledge, this is the first systematic investigation of the antioxidants from the leaves of M. hupehensis. The results indicated that the proposed method is an efficient technique to rapidly investigate antioxidants, especially for coeluted and minor compounds in a complex system.

“…As a continuous liquid–liquid partition chromatography method, complications of liquid–solid chromatography such as irreversible adsorption, low recovery, and deactivation or loss of activity of sample can be effectively eliminated due to lack of solid support matrix . 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 .…”

Section: Introductionmentioning

confidence: 99%

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

Huang

et al. 2019

Phytochemical Analysis

Self Cite

Introduction Potentilla kleiniana Wight et Arn is widely used as a herbal medicine to treat type 2 diabetes. However, detailed information about its active compounds is lacking. Objective To develop an efficient method for the rapid screening and separation of α‐glucosidase inhibitors from Potentilla kleiniana Wight et Arn. Methodology Potential α‐glucosidase inhibitors from Potentilla kleiniana Wight et Arn were rapidly screened out through ultrafiltration high‐performance liquid chromatography mass spectrometry (HPLC‐MS), and then followed by a target‐guided high‐speed counter‐current chromatography (HSCCC) separation using two‐phase solvent systems composed of n‐hexane/ethyl acetate/methanol/water (1:10:1:10, v/v/v/v and 1:10:5:6, v/v/v/v), and adopting increasing flow‐rate from 1.5 to 3.0 mL/min after 200 min. Their structures were identified by ultraviolet (UV), MS, proton nuclear magnetic resonance (1H‐NMR) and carbon‐13 (13C)‐NMR, and their α‐glucosidase inhibitory activities were assessed by in vitro assay. Results Five α‐glucosidase inhibitors including gallic acid (25.7 mg, 98.2%, 1), brevifolincarboxylic acid (9.86 mg, 95.3%, 2), ethyl evifolincarboxylate (13.26 mg, 97.6%, 3), 3,3′‐di‐O‐methylellagic acid‐4′‐O‐β‐d‐glucopyranoside (16.26 mg, 95.1%, 4), and 3,3′‐di‐O‐methylellagic acid (10.54 mg, 96.8%, 5) were successfully purified from 250 mg n‐butanol extract in a single run. Compounds 1, 2, 4 and 5 exhibited stronger α‐glucosidase inhibitory activities[half maximal inhibition concentration (IC50) values at 173.41 ± 6.35, 323.46 ± 8.08, 44.63 ± 2.50, and 20.73 ± 2.56 μM, respectively] than acarbose (IC50 value at 332.12 ± 5.52 μM, reference compound). Conclusions Notably, compounds 2–5 were reported in the Potentilla kleiniana Wight et Arn for the first time. The results indicated that the proposed method could be applied for the rapid screening and preparative separation of α‐glucosidase inhibitors from a complex matrix.