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

An anthocyanin and grape skin technology, applied in the field of food safety and food analytical chemistry, can solve the problems of in-depth research limitations, less standard anthocyanin samples, lack of anthocyanin color, etc., and achieve the effect of environmental friendliness and easy separation and operation

Inactive Publication Date: 2014-01-08
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] Example 1: Extraction of anthocyanins from grape skin

[0033] Frozen the harvested fresh and mature Cabernet Sauvignon wine grapes at -20°C. When the freezing is complete, take them out of the refrigerator and place them at room temperature to thaw the peels and peel them off (this method can be better for the peel and pulp Separation without sticking to the pulp). The extract is 95% ethanol (analytical pure) containing 1% hydrochloric acid (V / V), and is leached overnight at 30°C at a liquid-to-solid ratio of 1.5:1 by weight of extract: grape skin. The peel was filtered off and centrifuged at 6000 rpm for 10 min to remove the precipitate to obtain the extract.

Embodiment 2

[0034] Example 2: Processing of macroporous resin and preparation of sample loading

[0035] Soak the XAD-7HP macroporous resin with ethanol (analytical grade) first, and wash it every 12h until the eluate is colorless and transparent, and then wash it with deionized water until it is alcohol-free. Then it was soaked in 1mol / L NaOH for 12h, and then washed with deionized water to neutral pH. Change to 1mol / L HCl soaking for 12h, and finally wash with deionized water to neutral pH, and install the column for later use.

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

Embodiment 3

[0037] Example 3: Purification of anthocyanins in grape skins by macroporous resin

[0038] Pack the processed XAD-7HP macroporous resin on a column, and equilibrate with a solution of 0.1% hydrochloric acid (V / V) in 50% ethanol (diluted to 50% for analytical pure ethanol, V / V) for 0.5h. Sample injection at a flow rate of 0.5BV / h, eluted with a solution of 0.1% hydrochloric acid (V / V) in 50% ethanol (diluted to 50%, V / V for analytical pure ethanol) at a flow rate of 1BV / h, and collect the eluent (red) .

[0039] The characteristic absorption peak of anthocyanin is around 520nm, so the 520nm spectral absorption signal is used to monitor the change of anthocyanin content in the eluent. Impurity substances with benzene ring have strong spectral absorption at 280nm, so 280nm is used to monitor the content of impurities in the effluent.

[0040] Since the anthocyanins flowed out, samples were taken every 5 minutes to determine their absorbance values ​​at 520 nm and 280 nm. The collecti...

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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 Patents(China)
IPC IPC(8): C07H17/065C07H1/08C09B61/00
Inventor 韩富亮徐岩
Owner JIANGNAN UNIV
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