Characterization of Purified Red Cabbage Anthocyanins: Improvement in HPLC Separation and Protective Effect against H₂O₂-Induced Oxidative Stress in HepG2 Cells.

Sheng Fang,Liping Wang,Xianrui Liang,Fubin Lin,Daofeng Qu

doi:10.3390/molecules24010124

Abstract

In this study, the chemical profiles and antioxidant activities of red cabbage anthocyanin (RCA)-enriched extract are evaluated. The effects of column temperature on the HPLC resolution of the RCAs are studied. The HPLC resolutions became better as the column temperature increased from 20 °C–45 °C. An optimized HPLC condition was achieved at 45 °C and used for the quantification and qualification of the RCAs. The anthocyanins in the enriched powder are all derivatives of cyanidin (268 ± 2 μg/mg), mainly with 19% nonacylated, 51% monoacylated, and 31% diacylated structures with ferulic, sinapic, p-coumaric, and caffeic acids characterized by HPLC-MS. The RCA extracts markedly reduced intracellular oxidative stress production by H2O2 on HepG2 cells and consequently ameliorated cell apoptosis and improved viability. The analytical method and cellular antioxidant activity demonstration of the RCAs will greatly facilitate their functional applications.

Highlights

There is an increase in the application of natural anthocyanins as an alternative to synthetic pigments [1,2]
We found that the column temperature greatly influenced the separation of red cabbage anthocyanin (RCA) in a reversed-phase High-performance liquid chromatography (HPLC)
It is interesting to find that the temperature affects the resolution of RCAs in HPLC remarkably

Summary

Introduction

There is an increase in the application of natural anthocyanins as an alternative to synthetic pigments [1,2]. High-performance liquid chromatography (HPLC) is an extensively-used and powerful technique to separate and characterize anthocyanins in plants including RC [3,4,6,7]. Because of their similar structures, some anthocyanins in RC have a tendency to be co-eluted in an HPLC column [6], which makes it difficult to achieve a baseline separation. Zhang et al [7] found that, some peaks were isolated at the baseline after optimization in a reversed-phase HPLC, there were still other predominant RCAs eluted together either in a C18 or phenyl-hexyl column.

Methods

Results

Conclusion