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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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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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Acknowledgments

This work was supported by the National Scientific Foundation of China (no. 21005089), Natural Science Foundation of Hunan Province of China (no. 10JJ4006), the freedom explore Program of Central South University (201012200015), the Shenghua Yuying project of Central South University, the open fund of the State Key Laboratory of Powder Metallurgy, and aid program for Science and Technology Innovative Research Team (Chemicals of Forestry Resources and Development of Forest Products) in Higher Educational Institutions of Hunan Province.

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Authors and Affiliations

  1. State Key Laboratory of Powder Metallurgy, Central South University, Changsha, 410083, China

    Yu-Ping Zhang, Shu-Yun Shi & Xiang Xiong

  2. School of Chemistry and Chemical Engineering, Central South University, Changsha, 410083, China

    Yu-Ping Zhang, Shu-Yun Shi & Xiao-Qing Chen

  3. Key Laboratory of Hunan Forest Products and Chemical Industry Engineering, Jishou University, Zhangjiajie, 427000, China

    Mi-Jun Peng

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  1. Yu-Ping Zhang
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Correspondence to Shu-Yun Shi or Mi-Jun Peng.

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Zhang, YP., Shi, SY., Xiong, X. et al. 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. Anal Bioanal Chem 402, 2965–2976 (2012). https://doi.org/10.1007/s00216-012-5722-3

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