研究目的
Re-evaluating TiCl4 and UV assays for detection of vicine and convicine in high-throughput screening of immature and mature seeds of faba bean.
研究成果
The method of 274 nm UV absorption without an HPLC analysis offers more reliable analysis for detection of V-C in immature and mature seeds of faba bean. Its application in high throughput screening by 60 min agitation of immature seeds or mature seed flour in 2% trichloroacetic acid (TCA) allows quick screening of low V-C faba beans.
研究不足
The TiCl4 assay specificity is low, possibly because of the complexity of the plant metabolic system in immature seeds. The TiCl4 assay can and should be used together with the UV assay for selecting low V-C but high phenolic/tannin genotypes in cotyledons and/or whole seeds that are immature or mature.
1:Experimental Design and Method Selection:
The study re-evaluated two methodologies (TiCl4 assay and 274 nm UV absorption) for their application in detection of V-C in faba beans.
2:Sample Selection and Data Sources:
Mature and immature seeds of faba bean cultivars were obtained from the Crop Development Centre (CDC) of the University of Saskatchewan.
3:List of Experimental Equipment and Materials:
Titanium tetrachloride was obtained from Sigma-Aldrich Canada Co. V-C standards were purified from faba bean protein concentrate.
4:Experimental Procedures and Operational Workflow:
Seeds were extracted with 2% TCA, sonicated, and centrifuged. V-C was determined by comparing the V and C peak areas to the whole run area in chromatograms obtained from HPLC analysis.
5:Data Analysis Methods:
The amount of standard solutions injected on HPLC corresponding peak areas was used in quantification of V+C in HPLC runs and extracts.
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