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

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 natura...

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Published in:	Analytical and bioanalytical chemistry 2012-03, Vol.402 (9), p.2965-2976
Main Authors:	Zhang, Yu-Ping, Shi, Shu-Yun, Xiong, Xiang, Chen, Xiao-Qing, Peng, Mi-Jun
Format:	Article
Language:	eng
Subjects:	Analytical Chemistry Antioxidants Antioxidants - analysis Assaying Biochemistry Biphenyl Compounds - chemistry Characterization and Evaluation of Materials Chemistry Chemistry and Materials Science Chromatography, High Pressure Liquid - methods Comparative analysis Esters Flowers Flowers - chemistry Food Science High performance liquid chromatography Laboratory Medicine Liquid chromatography Methods Monitoring/Environmental Analysis Natural products Original Paper Picrates - chemistry Plant Extracts - analysis Pueraria - chemistry Screening Searching Spiking
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Summary:	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.
ISSN:	1618-2642 1618-2650