阅读说明：本技术 一种原花色素多聚体的制备方法及原花色素多聚体 (Preparation method of proanthocyanidin polymer and proanthocyanidin polymer ) 是由 杨海花 姜坤 叶焕峰 武永杰 于 2020-05-22 设计创作，主要内容包括：本发明公开了一种原花色素多聚体的制备方法,所述方法包括准备原料,所述原料为杨梅叶或葡萄籽；对原料进行预处理得到原料粉；将原料粉与丙酮水溶液混合得到混合溶液,然后向混合溶液中加入抗坏血酸,避光间歇震荡提取得到混合料液；对得到的混合料液进行抽滤得到滤液,将滤液与三氯甲烷混合收集上层液体；在收集到的上层液体中加入氯化钠,离心收集粗沉淀；对粗沉淀进行水洗,后经过冷冻干燥得到原花色素多聚体。本发明制备工艺简单,工艺条件温和,多聚体得率高且聚合度较高,制备得到的原花色素多聚体可用于原花色素结构及生物活性分析和应用,也可用于制备原花色素低聚体或金属螯合剂等。(The invention discloses a preparation method of proanthocyanidin polymer, which comprises the steps of preparing raw materials, wherein the raw materials are waxberry leaves or grape seeds; pretreating the raw materials to obtain raw material powder; mixing the raw material powder with an acetone aqueous solution to obtain a mixed solution, adding ascorbic acid into the mixed solution, and carrying out intermittent shaking extraction in a dark place to obtain a mixed material liquid; carrying out suction filtration on the obtained mixed feed liquid to obtain filtrate, mixing the filtrate with trichloromethane, and collecting upper-layer liquid; adding sodium chloride into the collected upper layer liquid, and centrifuging to collect a coarse precipitate; and washing the crude precipitate with water, and freeze-drying to obtain the proanthocyanidin polymer. The preparation method has the advantages of simple preparation process, mild process conditions, high polymer yield and higher polymerization degree, and the prepared proanthocyanidin polymer can be used for analyzing and applying the structure and the biological activity of proanthocyanidin and can also be used for preparing proanthocyanidin oligomer or metal chelating agent and the like.)

1. A method for preparing proanthocyanidin multimers, comprising the steps of:

s1, preparing a raw material, wherein the raw material is waxberry leaves or grape seeds;

s2, pretreating the raw materials to obtain raw material powder;

s3, mixing the raw material powder with an acetone aqueous solution to obtain a mixed solution, adding ascorbic acid into the mixed solution, and carrying out intermittent shaking extraction in a dark place to obtain a mixed material liquid;

s4, carrying out suction filtration on the obtained mixed feed liquid to obtain filtrate, mixing the filtrate with trichloromethane, and collecting upper-layer liquid;

s5, adding sodium chloride into the collected upper layer liquid, centrifuging and collecting coarse precipitate;

and S6, washing the crude precipitate with water, and freeze-drying to obtain proanthocyanidin polymer.

2. The method for preparing proanthocyanidin multimers of claim 1, wherein when the raw material is red bayberry leaves, the weight to volume ratio of raw material powder to aqueous acetone solution is: 1: 40-45 (g/mL).

3. The method for preparing proanthocyanidin multimers of claim 1, wherein when the starting material is grape seed, the weight to volume ratio of starting powder to aqueous acetone solution is: 1: 10-20 (g/mL).

4. The method for preparing proanthocyanidin multimers of claim 1, wherein the pre-treatment of step S2 comprises oven drying the raw material at 25-40 ℃ for 8-14 hours, followed by grinding and sieving through a 40-80 mesh sieve.

5. The method of preparing the proanthocyanidin multimer of claim 1, wherein the concentration of the aqueous acetone solution in step S3 is 50-80% (v/v).

6. The method of preparing the proanthocyanidin multimer of claim 1, wherein the concentration of ascorbic acid in step S3 is between 0.1% and 1% (w/v).

7. The method for preparing proanthocyanidin multimers according to claim 1, wherein the volume ratio of the filtrate to chloroform in step S4 is 1: 1-2.

8. The myrica rubra leaf proanthocyanidin polymer prepared by the preparation method of the proanthocyanidin polymer according to claim 1, wherein the average degree of polymerization of the myrica rubra leaf proanthocyanidin polymer is 24-31.

9. A grape seed proanthocyanidin multimer produced by the method of producing a proanthocyanidin multimer of claim 1, wherein the grape seed proanthocyanidin multimer has an average degree of polymerization of 6.2 to 7.3.

Technical Field

The invention belongs to the technical field of preparation of natural products, relates to food phytochemical components, and particularly relates to a preparation method of a proanthocyanidin polymer and the proanthocyanidin polymer.

Background

Proanthocyanidins are the most abundant manifestations of polyphenol substances in plants in addition to lignin, and also the most abundant flavonoid substances in human diet, which are abundant in plants, mainly present in plant epidermis, pericarp, seeds, etc.

The waxberry fruits and the grapes have extremely high added value, and the waxberry fruits and the grapes have been cultivated economically in China for many years and are relatively mature in industrialization. However, compared with waxberry fruits and grape pulp, the utilization rate of waxberry leaves and grape seeds is extremely low, and researches show that the proanthocyanidin content of the waxberry leaves is very high, the waxberry leaves mainly have prodelphinidin in structure, and the grape seeds also contain a large amount of proanthocyanidins which mainly are proanthocyanidins with different polymerization degrees. The proanthocyanidin polymer is proanthocyanidin with polymerization degree of 5 or more, that is, containing 5 or more flavan-3-ol monomers in structure, and the polymer with polymerization degree of 2-4 is oligomer. Many studies on proanthocyanidins are still conducted on oligomers, and although the content of polymers in plants is considerable, sufficient attention and intensive studies have not been obtained yet, mainly because proanthocyanidin polymers have too large molecular weights to pass through epithelial cells, and thus cannot be directly absorbed and utilized by organisms to exert an active effect, and further, the polymers have extremely complicated structures and great analysis difficulty.

The common preparation methods of proanthocyanidins at present mainly comprise a solvent extraction method, a microwave-assisted extraction method, a supercritical fluid extraction method and the like. The solvent extraction method has low extraction efficiency and is time-consuming; the microwave-assisted extraction method has limited processing capacity, is not suitable for large-scale extraction, and can damage the structure of proanthocyanidins to a certain extent due to long-time extraction. Supercritical CO₂The extraction method has complex operation and high cost, and simultaneously, an entrainer is required to be added for use. Most importantly, the preparation methods are mainly used for extraction preparationProanthocyanidin oligomers are prepared rather than polymers. Compared with proanthocyanidin oligomer, a plant has a larger amount of polymer, and in order to deeply research, develop and utilize the polymer, the invention of a simple extraction preparation method with high proanthocyanidin polymer yield is necessary.

Disclosure of Invention

Aiming at the defects or shortcomings of the prior art, the invention aims to provide a preparation method of proanthocyanidin polymer and the proanthocyanidin polymer so as to solve the technical problem that the preparation method of the proanthocyanidin polymer is lacked in the prior art.

In order to achieve the purpose, the invention adopts the following technical scheme:

a method for preparing proanthocyanidin multimers comprises the steps of:

s1, preparing a raw material, wherein the raw material is waxberry leaves or grape seeds;

s2, pretreating the raw materials to obtain raw material powder;

s3, mixing the raw material powder with an acetone aqueous solution to obtain a mixed solution, adding ascorbic acid into the mixed solution, and carrying out intermittent shaking extraction in a dark place to obtain a mixed material liquid;

s4, carrying out suction filtration on the obtained mixed feed liquid to obtain filtrate, mixing the filtrate with trichloromethane, and collecting upper-layer liquid;

s5, adding sodium chloride into the collected upper layer liquid, centrifuging and collecting coarse precipitate;

and S6, washing the crude precipitate with water, and freeze-drying to obtain proanthocyanidin polymer.

Specifically, when the raw material is waxberry leaves, the weight volume ratio of the waxberry leaf powder to the acetone aqueous solution is as follows: 1: 40-45 (g/mL).

Specifically, when the raw material is grape seeds, the weight volume ratio of the grape seed powder to the acetone aqueous solution is as follows: 1: 10-20 (g/mL).

Specifically, the pretreatment in the step S2 includes drying the raw materials at 25-40 ℃ for 8-14 h, and then grinding and sieving the raw materials with a 40-80 mesh sieve.

Specifically, the concentration of the acetone aqueous solution in step S3 is 50 to 80% (v/v).

Specifically, the concentration of the ascorbic acid in step S3 is 0.1% to 1% (w/v).

Specifically, the volume ratio of the filtrate to the chloroform in the step S4 is 1: 1-2.

The invention also discloses the waxberry leaf proanthocyanidin polymer prepared by the preparation method, and the average polymerization degree of the waxberry leaf proanthocyanidin polymer is 24-31.

The invention also discloses the grape seed proanthocyanidin polymer prepared by the preparation method, wherein the average polymerization degree of the grape seed proanthocyanidin polymer is 6.2-7.3.

Compared with the prior art, the invention has the beneficial effects that:

compared with the traditional preparation method, the method can be used for preparing the proanthocyanidin polymers of different varieties of plants, and has the advantages of simple preparation process, mild process conditions, high polymer yield, higher polymerization degree of the obtained polymers and batch production.

Drawings

FIG. 1 is an HPLC chart of the polymeric acid degradation product of proanthocyanidins from Myrica rubra leaves prepared in example 1; wherein EGCG-ph is an addition product of epigallocatechin gallate phloroglucinol.

Fig. 2 is an HPLC diagram of the acid degradation product of the grape seed proanthocyanidin multimer prepared in example 2; wherein C-ph, EC-ph and CG-ph are catechin phloroglucinol adduct, epicatechin phloroglucinol adduct and catechin gallate phloroglucinol adduct respectively.

Detailed Description

The following embodiments of the present invention are provided, and it should be noted that the present invention is not limited to the following embodiments, and all equivalent changes based on the technical solutions of the present invention are within the protection scope of the present invention.

Unless otherwise specified, materials, reagents and the like used in the following examples are commercially available reagents and materials.