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Method for separating and preparing five high-purity anthocyanidin monomers from grape skins

An anthocyanin and grape skin technology, which is applied in the fields of food safety and food analytical chemistry, can solve the problems of few anthocyanin standard samples, limited in-depth research, high price, etc., and achieves the effect of simple and easy separation operation and environmental friendliness.

Inactive Publication Date: 2011-11-02
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] According to research reports, more than 500 kinds of anthocyanins have been found, but the standard samples of anthocyanins available in the market are few and expensive
In the study of wine anthocyanins, the quantification mostly uses a standard sample of Mv3-glu to carry out relative quantification of all anthocyanins, and absolute quantification cannot be achieved
The lack of anthocyanin standard samples limits the in-depth research on anthocyanin color, taste, physiological activity, etc.
Two kinds of anthocyanins, Mv3-acet-glu and Mv3-coum-glu, are abundant in grape skins, but there are still no standard samples for purchase due to the difficulty of separation

Method used

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  • Method for separating and preparing five high-purity anthocyanidin monomers from grape skins
  • Method for separating and preparing five high-purity anthocyanidin monomers from grape skins
  • Method for separating and preparing five high-purity anthocyanidin monomers from grape skins

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment 1: Extraction of grape skin anthocyanins

[0033] Freeze the harvested fresh and ripe Cabernet Sauvignon wine grapes at -20°C. After the freezing is complete, take them out of the refrigerator and place them at room temperature, thaw the peel and peel off the peel (this method can better preserve the peel and pulp. separation without sticking the pulp). The extract is 95% ethanol (analytically pure) containing 1% hydrochloric acid (V / V), and it is leached overnight at 30°C according to the liquid-to-solid ratio of extract:grape skin weight ratio 1.5:1. Filter off the peel and centrifuge at 6000r for 10min to remove the precipitate to obtain the extract.

Embodiment 2

[0034] Embodiment 2: the processing of macroporous resin and the preparation of loading sample

[0035] Soak the XAD-7HP macroporous resin with ethanol (analytical grade) first, and wash it every 12 hours until the eluate is colorless and transparent, and then wash it with deionized water until there is no alcohol. Then soak with 1mol / L NaOH for 12h, and then wash with deionized water to neutral pH. Replace with 1mol / L HCl to soak for 12 hours, and finally wash with deionized water to neutral pH, and pack it into a column for later use.

[0036]The extract obtained from the treatment in Example 1 was vacuum rotary evaporated at 30° C. to remove ethanol, and then vacuum freeze-dried to obtain a crude anthocyanin extract. The crude extract was dissolved in 50% ethanol (analytical grade diluted to 50%, V / V) until the solution was saturated, centrifuged at 8000r for 6min, filtered through a 0.45μm organic microporous membrane and prepared for sample injection.

Embodiment 3

[0037] Embodiment 3: Macroporous resin purifies anthocyanins in grape skin

[0038] Pack the treated XAD-7HP macroporous resin into a column, and equilibrate with 50% ethanol (analytical pure ethanol diluted to 50%, V / V) solution containing 0.1% hydrochloric acid (V / V) for 0.5h. Inject at a flow rate of 0.5BV / h, elute with a solution of 50% ethanol (analytical pure ethanol diluted to 50%, V / V) containing 0.1% hydrochloric acid (V / V) at a flow rate of 1BV / h, and collect the eluate (red) .

[0039] The characteristic absorption peak of anthocyanin is around 520nm, so the content change of anthocyanin in the eluate is monitored by the spectral absorption signal at 520nm. Impurities with benzene rings have strong spectral absorption at 280nm, so 280nm is used to monitor the content of impurities in the effluent.

[0040] Samples were taken every 5 minutes to measure the absorbance values ​​at 520nm and 280nm since the anthocyanins flowed out, and the collection time was taken as...

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Abstract

The invention discloses a method for separating and preparing five high-purity anthocyanidin monomers from grape skins, relating to the preparation of natural pigments and belonging to the fields of food safety and food analytic chemistry. The method comprises the following steps of: firstly, carrying out anthocyanidin digestion on carbernet sauvignon grape skins to obtain crude extractive of anthocyanidin; then purifying the crude extractive by using XAD-7HP macroporous resin, and removing most of moisture absorption components such as saccharides, inorganic salts, phlegmatic temperament and the like in the crude extractive; and carrying out preparative liquid phase separation on the extractive of anthocyanidin subjected to the preliminary purification to obtain the high-purity monomers of the anthocyanidin. The method has the advantages of only using alcohol and acid as a solvent in stages of extraction, purification and separation in the preparation of the anthocyanidin and being environment-friendly, simple, convenient and feasible. The five anthocyanidin monomers obtained after purification are respectively delphinidin glucoside, 3'-methyl delphinidin glucoside, dimethyl delphinidin glucoside, dimethyl delphinidin acetylation glucoside, and dimethyl delphinidin trans-tonka-bean acetylation glucoside, the purities of which are sequentially 99.9%, 99.9%, 99.9%, 99.9%, 91.7%, 95.5%.

Description

technical field [0001] The invention discloses a method for separating and preparing five high-purity anthocyanin monomers from grape skins, relates to the preparation of natural pigments, and belongs to the fields of food safety and food analytical chemistry. Background technique [0002] Anthocyanin is a natural pigment that is beneficial to the human body. It can be used as food coloring and is widely used in food. Anthocyanins help the body fight inflammation, inhibit lipid peroxidation, and improve blood circulation. Natural anthocyanins in grapes are mainly found on the skins, which endow the wine with a bright red color during the brewing process. Grapes are one of the fruits with the largest annual production in the world, and most of them are used in winemaking. In Vitis vinifera, Delphinidin-3-glucoside (Dp3-glu), 3'methyl delphinidin-glucoside (Petunidin-3-glucoside, Pt3-glu), delphinidin-3 Glucoside (Malvidin-3-glucoside, Mv3-glu) and its acetylation (Malvidin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07H17/065C07H1/08C09B61/00
Inventor 韩富亮徐岩
Owner JIANGNAN UNIV
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