CN108558971A - A kind of preparation method of roselle anthocyanin - Google Patents

Abstract

The invention discloses a method for preparing roselle anthocyanins, belonging to the field of biochemical products. The present invention uses roselle calyx as raw material, and its technological process includes: after the roselle calyx is pulverized, high-purity anthocyanin is obtained by using a combined technology of pretreatment solution impurity removal→extraction extraction→membrane separation→macroporous resin separation. color price , can be used in the fields of food, health products and medicine. The invention has simple process and low cost, and can be used for large-scale production of high-purity anthocyanins.

Description

A kind of preparation method of roselle anthocyanin

technical field

The invention relates to the field of biochemical products, in particular to a method for preparing roselle anthocyanins.

Background technique

Anthocyanins are a class of water-soluble pigments that widely exist in the plant kingdom and belong to flavonoids. The benzene ring of anthocyanin contains multiple phenolic hydroxyl groups, has a highly conjugated structure, and has oxycations, which have strong antioxidant and free radical scavenging capabilities. Free radicals play an important role in human metabolism, but if the human body produces too many free radicals or metabolic disorders, it may induce gene mutation, protein denaturation and lipid peroxidation, and then damage various cells, causing oxidative stress diseases. There are many reports that anthocyanins have good effects on the prevention and treatment of cardiovascular diseases, hypertension, cancer and diabetes.

Roselle (Hibiscus sabdariffa) is an annual tropical plant of the Malvaceae family. It originated in Africa and then spread to all over the world. It is widely planted in Fujian, Guangdong, Guangxi, and Yunnan in my country. Roselle is rich in various bioactive substances such as flavonoids, anthocyanins, organic acids and polysaccharides, and has good medical value. Roselle is a traditional medicinal and edible plant. The calyx is succulent and rich in anthocyanins. It is an important plant for extracting anthocyanins.

The traditional extraction process of anthocyanins mostly adopts leaching, evaporation concentration, solvent purification and other processes, which have problems such as high energy consumption, large pigment damage, and low product purity.

Contents of the invention

The object of the present invention is to provide a method for preparing roselle anthocyanins aiming at the deficiencies and shortcomings of the traditional technology. The present invention adopts the combination technology of pretreatment solution impurity removal→extraction liquid extraction→membrane separation→macroporous resin separation to obtain high-purity roselle anthocyanins, the main components of which are delphinidin-3-O-sambaribioside and cyanidin-3-O-sambranbioside, its color value .

To achieve the above object, the present invention adopts the following technical solutions:

(1) Impurity removal of pretreatment solution: take roselle calyx, crush it to a particle size of 100-1000 μm, mix the crushed material with the pretreatment solution, and the ratio of solid to liquid is 1:5-1:100 (g/mL), The temperature is 4-60°C, stirred for 0.5-10 hours, and the solid-liquid separation collects the filter residue. The obtained filter residue is repeated for 1-6 times, and the pretreatment solution is recovered and reused;

(2) Extraction: Mix the filter residue obtained in step (1) with the extract, the ratio of solid to liquid is 1:5-1:100 (m/v), stir and extract for 0.5-6h, the temperature is 4-70°C, Repeat the extraction 1-6 times, and combine the supernatant to obtain anthocyanin extract;

(3) Membrane separation: remove macromolecular impurities in the anthocyanin extract obtained in step (2) by membrane separation;

(4) Macroporous adsorption resin separation: the anthocyanin extract obtained in step (3) is subjected to specific adsorption-desorption using a macroporous adsorption resin to remove small molecular impurities to obtain a purified roselle anthocyanin liquid.

The pretreatment solution described in step (1) is formed by mixing an organic solvent and water at a ratio of 10:0.1-10 (V/V), wherein the organic solvent is petroleum ether, solvent naphtha, ether, ethyl acetate, n-hexane, One or more mixtures of cyclohexane.

The extract described in step (2) is obtained by mixing an organic solvent and water at a ratio of 10:0.1-10 (V/V), wherein the organic solvent is one or more of methanol, ethanol, propanol, acetone, and acetonitrile kind of mix.

In step (3), the membrane separation method is used to remove macromolecular impurities in the anthocyanin extract, and the membrane materials used are polyolefins, polysulfones, inorganics, fluorine-containing polymer materials, polyamide cellulose One or more mixtures of crustacean or crustacean membrane materials.

In step (4), macroporous resins are used for specific adsorption-desorption to remove small molecular impurities, wherein the macroporous resin skeleton used is one or more of styrene, acrylic acid, phenolic or epoxy.

The beneficial effects of the present invention are: the process is simple, the cost is low, and it can be used for large-scale production of high-purity anthocyanins. , can be used in the fields of food, health products and medicine.

Description of drawings

Fig. 1 is a process flow diagram for the preparation of high-purity roselle anthocyanins;

Fig. 2 is high-purity roselle anthocyanin high performance liquid phase chromatogram;

Figure 3(a) is the UV-visible spectrum of delphinidin-3-O-sambioside; Figure 3(b) is the UV-visible spectrum of cyanidin-3-O-sambrandioside .

Detailed ways

The present invention will be further described below in conjunction with specific examples, but the present invention is not limited to these examples.

Example 1

Grind roselle calyx to a particle size of 100 μm, weigh 1000 g of coarse pollen, add 5 L of pretreatment solution (petroleum ether: water = 10:1, V/V), stir at 30 °C for 4 h, separate solid-liquid to take filter residue, and obtain Repeat the treatment of the filter residue 3 times; add 10L of extract (40% ethanol solution) to the obtained filter residue, extract at 40°C for 2 hours, repeat the extraction 2 times, combine the supernatant to obtain anthocyanin extract; use polysulfone ultrafiltration membrane Remove macromolecular impurities in the extract; the solution after membrane separation is dynamically adsorbed by HZ-802 macroporous adsorption resin, anthocyanins are eluted with 60% ethanol solution, the eluent is rotary evaporated to remove ethanol, and spray-dried to obtain high-purity flowers and colors glycosides. The product is dark red, the color price is 152.5.

Example 2

Grind the dried calyx of roselle to a particle size of 500 μm, weigh 500 g of the raw material, add 5 L of pretreatment solution (ethyl acetate: water = 10:2, V/V), stir at 30 °C for 3 h, separate the solid and liquid to take the filter residue, The obtained filter residue was repeatedly treated 3 times; 10L of 60% methanol solution was added to the obtained filter residue, extracted at 60°C for 1 hour, and the extraction was repeated 3 times, and the supernatant was combined to obtain anthocyanin extract; nylon ultrafiltration membrane was used to remove the Macromolecular impurities; the solution after membrane separation is dynamically adsorbed by X-5 macroporous adsorption resin, the anthocyanin sample is eluted with 80% ethanol, the ethanol is removed by rotary evaporation, and high-purity anthocyanin is obtained by spray drying. The product is red, its color value is 154.7.

Example 3

Grind the dried roselle calyx to a particle size of 1000 μm, weigh 250 g of the raw material, add 5 L of pretreatment solution (n-hexane: water = 10:5, V/V), stir at 40 °C for 6 h, separate the solid and liquid to take the filter residue; Repeat the treatment of the filter residue twice; add 10 L of 60% acetone solution to the obtained filter residue, extract in the dark at 20°C for 4 hours, repeat the extraction 4 times, and combine the supernatants to obtain anthocyanin extract. Polytetrafluoroethylene ultrafiltration membrane is used to remove macromolecular impurities in the extract; the solution after membrane separation is dynamically adsorbed by D101 macroporous adsorption resin to contain anthocyanin, 90% ethanol solution is used to elute the anthocyanin sample, and the eluent is rotary evaporated Ethanol is removed, and high-purity anthocyanins are obtained by spray drying. The product is red, its color value for 151.93.

The HPLC-MS analysis results of the prepared high-purity roselle anthocyanins are shown in Table 1.

Table 1 HPLC-MS analysis results of high-purity roselle anthocyanins

The above descriptions are only preferred embodiments of the present invention, and all equivalent changes and modifications made according to the scope of the patent application of the present invention shall fall within the scope of the present invention.

Claims (7)

1. a kind of preparation method of roselle anthocyanin, it is characterised in that：Include the following steps：Use Roselle calyx for original Material, after crushed, then preprocessed solution removal of impurities → extract liquor extraction → UF membrane → macroporous absorbent resin detaches group technology Obtain roselle anthocyanin refined solution, color value。

2. the preparation method of roselle anthocyanin according to claim 1, it is characterised in that：Specifically include following steps：

（1）Preprocessing solution cleans：Take Roselle calyx, pulverization process to 100-1000 μm of grain size, will crush resulting material with Preprocessing solution mixes, and temperature is 4-60 DEG C, stir process 0.5-10h, is separated by solid-liquid separation and collects filter residue, and gained filter residue repeats above-mentioned Processing 1-6 times, preprocessing solution recycling are reused；

（2）Extract liquor extracts：By step（1）Obtained filter residue is mixed with extract liquor, stirring extraction 0.5-6h, temperature 4-70 DEG C, extraction 1-6 times is repeated, merges supernatant and obtains anthocyanin extracting solution；

（3）UF membrane：Using membrane separation process removal step（2）Big molecular impurity in obtained anthocyanin extracting solution；

（4）Macroporous absorbent resin detaches：Using macroporous absorbent resin to step（3）Obtained anthocyanin extracting solution carries out specificity Absorption-desorption removes small molecular weight impurity, obtains roselle anthocyanin refined solution.

3. the preparation method of roselle anthocyanin according to claim 1, it is characterised in that：Step（1）Described in it is pre- Processing solution is that organic solvent presses 10 with water:0.1-10（V/V）It mixes, wherein organic solvent is petroleum ether, solvent naphtha, second One or more of ether, ethyl acetate, n-hexane, hexamethylene mix.

4. the preparation method of roselle anthocyanin according to claim 1, it is characterised in that：Step（1）In, after crushing Material is mixed with preprocessing solution, solid-liquid ratio 1:5-1:100（g/mL）；Step（2）In, filter residue is mixed with extract liquor, feed liquid Than being 1:5-1:100（g/mL）.

5. the preparation method of roselle anthocyanin according to claim 1, it is characterised in that：Step（2）Described in extraction It is organic solvent and water by 10 to take liquid:0.1-10（V/V）Mix, wherein organic solvent be methanol, ethyl alcohol, propyl alcohol, acetone, One or more of acetonitrile mixes.

6. the preparation method of roselle anthocyanin according to claim 1, it is characterised in that：Step（3）In, described adopts With membrane separation process remove anthocyanin extracting solution in big molecular impurity, membrane material used be polyolefins, polysulfones, inorganic, One or more of fluoro-containing macromolecule material, polyamide fiber element class or shell-fish membrane material mixing material.

7. the preparation method of roselle anthocyanin according to claim 1, it is characterised in that：Step（4）In, using macropore Adsorb resin carry out specific adsorption-desorption, remove small molecule class impurity, used in macroreticular resin skeleton be styrene, One or more of acrylic acid, phenolic aldehyde or epoxy.