Screening of Phenolic Antioxidant Components in Polygonum perfoliatum L. Using LC×LC-ECD and UPLC-Orbitrap-MS/MS
- 1School of Pharmacy, Zunyi Medical University, Zunyi 563000, China
- 2School of Basic Medical Sciences, Zunyi Medical University, Zunyi 563000, China
Abstract
Polygonum perfoliatum L. is a medicinal plant rich in phenolic and flavonoid compounds, but severe peak overlap in one-dimensional liquid chromatography (1D-LC) hinders the separation of antioxidants. We developed an offline comprehensive two-dimensional liquid chromatography-electrochemical detection (LC×LC-ECD) platform with high orthogonality and sensitive electrochemical response for screening of antioxidants. Identification was performed using ultra-high-performance liquid chromatography-high-resolution tandem mass spectrometry (UHPLC-Orbitrap-MS/MS), and quantification was performed using high-performance liquid chromatography with electrochemical detection (HPLC-ECD). A total of 63 antioxidants were annotated, mainly phenolic acids and flavonoids, with caffeoyl and quercetin derivatives most abundant. The first-dimension fractions exhibited radical scavenging capacity, and their peak areas were positively correlated with the activity, with correlation coefficients of 0.9697 (p < 0.001) for 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) and 0.9688 (p < 0.001) for 1,1-diphenyl-2-picrylhydrazyl (DPPH). The 11 major compounds were quantitatively analyzed. Tartroylcaffeic acid and quercetin 3-O-β-D-glucuronide predominated, with average contents of 5101.7–29,296.2 µg/g and 3876.9–21,535.2 µg/g, respectively. Multivariate statistical analysis indicated that geographical origin influenced compound profiles, highlighting the need for source-specific evaluation. This offline LC×LC-ECD strategy overcomes the separation bottlenecks of 1D-LC in complex plant extracts and provides a new paradigm for systematic antioxidant screening and activity-guided separation.
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BibTeX
@article{pan2026screening,
title = {{Screening of Phenolic Antioxidant Components in Polygonum perfoliatum L. Using LC×LC-ECD and UPLC-Orbitrap-MS/MS}},
author = {Wei Pan and Xing-Rui Li and Hai-Yan Gong and Zheng-Yan Tan and Pei Xu and Piao Wang and Xing-Qin Luo and Cheng Yang and Ya-Ping Zhou and Rong-Xiang Chen},
journal = {Molecules},
year = {2026},
month = oct,
doi = {10.3390/molecules31203612},
url = {https://www.mdpi.com/1420-3049/31/20/3612},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Screening of Phenolic Antioxidant Components in Polygonum perfoliatum L. Using LC×LC-ECD and UPLC-Orbitrap-MS/MS
AU - Wei Pan
AU - Xing-Rui Li
AU - Hai-Yan Gong
AU - Zheng-Yan Tan
AU - Pei Xu
AU - Piao Wang
AU - Xing-Qin Luo
AU - Cheng Yang
AU - Ya-Ping Zhou
AU - Rong-Xiang Chen
PY - 2026
DA - 2026/10/01
JO - Molecules
DO - 10.3390/molecules31203612
UR - https://www.mdpi.com/1420-3049/31/20/3612
AB - Polygonum perfoliatum L. is a medicinal plant rich in phenolic and flavonoid compounds, but severe peak overlap in one-dimensional liquid chromatography (1D-LC) hinders the separation of antioxidants. We developed an offline comprehensive two-dimensional liquid chromatography-electrochemical detection (LC×LC-ECD) platform with high orthogonality and sensitive electrochemical response for screening of antioxidants. Identification was performed using ultra-high-performance liquid chromatography-high-resolution tandem mass spectrometry (UHPLC-Orbitrap-MS/MS), and quantification was performed using high-performance liquid chromatography with electrochemical detection (HPLC-ECD). A total of 63 antioxidants were annotated, mainly phenolic acids and flavonoids, with caffeoyl and quercetin derivatives most abundant. The first-dimension fractions exhibited radical scavenging capacity, and their peak areas were positively correlated with the activity, with correlation coefficients of 0.9697 (p < 0.001) for 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) and 0.9688 (p < 0.001) for 1,1-diphenyl-2-picrylhydrazyl (DPPH). The 11 major compounds were quantitatively analyzed. Tartroylcaffeic acid and quercetin 3-O-β-D-glucuronide predominated, with average contents of 5101.7–29,296.2 µg/g and 3876.9–21,535.2 µg/g, respectively. Multivariate statistical analysis indicated that geographical origin influenced compound profiles, highlighting the need for source-specific evaluation. This offline LC×LC-ECD strategy overcomes the separation bottlenecks of 1D-LC in complex plant extracts and provides a new paradigm for systematic antioxidant screening and activity-guided separation.
SN - 1420-3049
ER -