Comparative evaluation of three methods based on high-performance liquid chromatography analysis combined with a 2,2'-diphenyl-1-picrylhydrazyl assay for the rapid screening of antioxidants from Pueraria lobata flowers |
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Authors: | Zhang Yu-Ping Shi Shu-Yun Xiong Xiang Chen Xiao-Qing Peng Mi-Jun |
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Institution: | (1) State Key Laboratory of Powder Metallurgy, Central South University, Changsha, 410083, China;(2) School of Chemistry and Chemical Engineering, Central South University, Changsha, 410083, China;(3) Key Laboratory of Hunan Forest Products and Chemical Industry Engineering, Jishou University, Zhangjiajie, 427000, China; |
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Abstract: | Traditional activity-guided fractionation of natural products is a time-consuming, labor intensive, and expensive strategy,
which cannot compete with high-throughput and rapid screening of natural products. Therefore, more efficient approaches are
necessary for searching active compounds from natural products. Three main methods based on high-performance liquid chromatography
(HPLC) analysis combined with 2,2′-diphenyl-1-picrylhydrazyl (DPPH) assay, DPPH spiking HPLC analysis, on-line post-column
HPLC-DPPH analysis, and HPLC-based DPPH activity profiling, were then developed for the rapid screening of antioxidants from
complex mixtures. In the present study, a comparative study of these three methods has been conducted to identify antioxidants
from an ethyl acetate fraction of Pueraria lobata flowers. The parameters in HPLC analysis and DPPH assay were optimized. The results indicated that all three methods could
achieve similar information with regard to antioxidants, without the need for preparative isolation techniques. However, there
were differences in instrumental set-up, sensitivity, and efficiency. DPPH spiking HPLC analysis seemed to be more sensitive
and effective with simpler instrumental set-up and easier operation, which could also detect the total antioxidant capacity
of color complexes. Eighteen antioxidants were tentatively screened and identified from P. lobata flowers by DPPH spiking HPLC-MS/MS. Among them, ten compounds including one new compound were first isolated from P. lobata flowers, and the DPPH radical scavenging activity of the new compound was reported for the first time. |
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