High-throughput micro plate assays for screening flavonoid content and DPPH-scavenging activity in sorghum bran and flour

J Sci Food Agric. 2012 Aug 30;92(11):2326-31. doi: 10.1002/jsfa.5633. Epub 2012 Mar 14.

Abstract

Background: Sorghum possesses phenolic compounds that are health-promoting constituents of the grain. There are approximately 40 000 sorghum accessions, many of which have not been evaluated for the grain's health-promoting potential. Conventional methods for measuring total phenolic content, flavonoid content and 2,2-diphenyl-1-picrylhydrazyl (DPPH)-scavenging capacity are time-consuming and labour-intensive, resulting in low overall throughput. The objective of this study was to develop a high-throughput screening assay for large sorghum sample sets to determine flavonoid and phenolic content and to modify existing DPPH and total phenolic assays.

Results: The 96-well assays exhibited a correlation of > 0.9 with the conventional assays. The 96-well assays allowed for up to 64 samples to be run per day compared with 20-24 samples (depending on the test) for the conventional methods. The 96-well assays had excellent accuracy (97.65-106.16% recovery), precision (1.06-8.28% coefficient of variation (CV)) and reproducibility (1.32-2.13% CV inter-day and 1.36-2.09% CV intra-day).

Conclusion: The high-throughput 96-well plate method proved to be as robust and reproducible as the conventional method for determining total phenolic content, flavonoid content and DPPH-scavenging capacity in either sorghum bran or flour.

MeSH terms

  • Crops, Agricultural / chemistry
  • Dietary Fiber / analysis
  • Drug Discovery*
  • Flavonoids / analysis*
  • Flavonoids / chemistry
  • Food Additives / analysis
  • Food Additives / chemistry
  • Food Handling*
  • Food, Fortified
  • Free Radical Scavengers / analysis*
  • Free Radical Scavengers / chemistry
  • Functional Food / analysis
  • High-Throughput Screening Assays
  • Phenols / analysis
  • Phenols / chemistry
  • Plant Extracts / chemistry
  • Plant Extracts / isolation & purification
  • Reproducibility of Results
  • Seeds / chemistry*
  • Sorghum / chemistry*
  • Species Specificity

Substances

  • Dietary Fiber
  • Flavonoids
  • Food Additives
  • Free Radical Scavengers
  • Phenols
  • Plant Extracts