Extraction, Separation and Detection of Stilbenes in the Skin of ‘Kyoho’ Grape
Date Issued
2008
Date
2008
Author(s)
Tu, Yu-Fen
Abstract
The objects of this research were use the skin of grape (Vitis vinifera × Vitis labrusca ‘Kyoho’) to study the influence of extract solvents, sample purification and separation program of HPLC on extraction and determination of trans-piceid and trans-resveratrol.esults showed, using methanol of 0.1% hydrochloric acid as a extract solvent got the most amount of trans-piceid, 75% acetone extracts the most trans-resveratrol, and ethyl acetate was unsuitable to be used for extracting and purifying these two compounds. Using 100% methanol to extract trans-piceid and trans-resveratrol in the grape skin without purification, via HPLC separated and detected by a UV detector ( λ=320 nm ), and obtained relatively standard deviation of data (RSD) between 4.19~5.29 %.sing ethyl acetate : water : acetic acid =15 : 5 : 1 (v/v/v) as a development solvent to carry out thin-layer chromatography could separate trans-piceid and trans-resveratrol from the extracts, but lost 50%. Proceeding solid phase extraction (SPE) by Amberlite XAD-2 to purify extracts, as trans-piceid and trans-resveratrol dissolving in 10% methanol, the adsorbent ratio in Amberlite XAD-2 was best; and using 80% acetonitrile to elute, had the best efficiency to recover trans-piceid and trans-resveratrol absorbed on the Amberlite XAD-2. The study of HPLC using a C18 column (Thermo，Hypersil-100 C18，5μm, 250 mm× 4.6 mm) to separate trans-piceid, trans-resveratrol and other compounds in extracts of ‘Kyoho’ grape skin and Polygonum cuspidatum. Results of separation detected by a UV detector ( λ=320 nm ). Results of tests showed, while analysing trans-piceid and trans-resveratrol in grapes skin or Polygonum cuspidatum extracts at the same time, using 40% acetonitrile as mobile phase set at 0.6 ml/min isocratic could not separate well, and the chromatogram could not be determined. Contrarily, using acetonitrile and water as mobile phase set at 1 ml/min gradient made better separation than isocratic. To compare divers gradient steepness, gradient elution increased acetonitrile linearly from 10% to 85% over 72 min (GS = 1.04) with a flow rate of 1 ml/min made the best resolution of trans-piceid and trans-resveratrol peaks from grapes skin or Polygonum cuspidatum extracts. As column temperature increased there was a decrease in retention time of both compounds, but the theoretical plates decreased and the peaks widened resulted in resolution decreased. In order to detect well, the column set at 30℃. To develop a quick and accurate method through these results of extraction, purification, separation and detection make analyzing trans-piceid and trans-resveratrol in horticulture crops easy.
Subjects
stilbenes
trans-piceid
trans-resveratrol
grapes
Type
thesis
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