TWI396582B - Preparation method of high content quercus algae zeaxanthin antioxidant health care raw material - Google Patents

Description

Preparation method of high content of chlorella zeaxanthin antioxidant health raw material

The invention is mainly applied to the technology for preparing high-content phycocyanin zeaxanthin antioxidant health raw materials.

Nannochloropsis oculata, also known as Chlorella sp., Chlorella, is a spherical or slightly elliptical unicellular alga. The classification belongs to the genus Jinzao, the true eye point algae, the true eye, the genus Algae, and the genus Quercus. It is rich in EPA, oils and carotenoids and is often used as aquafeed.

Most microalgae are rich in carotenoids, which play an important role in plants - protecting plants from photo-oxidative. Carotenoids are also one of the essential nutrients for the human body. They have antioxidant and anti-cancer effects, and can also reduce the chance of cardiovascular disease. Carotenoids are part of the antioxidant defense system in human organs. Because carotenoids have antioxidant properties, carotenoids are susceptible to cleavage reactions in the presence of light, oxygen or heat. Most carotenoids have a maximum absorption of about 450 nm, and the ability to filter blue light is seen as a mechanism to protect the macula lutea against photooxidation. More and more studies have shown that carotenoids can protect the skin against the effects of photooxidation.

Carotenoids are mainly composed of eight isoprenes, forming a sequence of conjugated double bonds between molecules. Carotenoids can be divided into two broad categories depending on the functional group:

(a) Carotenes - oxygen-free functional groups. For example: β-carotene and lycopene.

(b) xanthophylls - oxygen-containing functional groups (such as -OH and =O). For example: zeaxanthin, lutein, canthaxanthin, astaxanthin and capsanthin. Generally, freshwater aquaculture has a zeaxanthin content of less than 0.2 mg per gram of microalgae.

However, the extraction of carotenoids and chlorophyll from microalgae by conventional methods often requires more than one extraction step by means of an organic solvent, however this is prohibited during the food addition process. In addition, the separation of carotenoids from chlorophyll requires more process steps. Therefore, a faster process method suitable for food manufacturing becomes important.

Supercritical carbon dioxide extraction, with high binding force of carbon dioxide and solvent, the extract has no solvent residue, and can operate at a lower temperature, does not provide excess heat, thereby reducing carotenoid oxidative cleavage. As for the zeaxanthin purification technology, there are currently no related patents in China. The applicant of the case filed an application for the invention patent number No. 098136811 "Supercritical fluid anti-solvent microalgae carotenoids and purified zeaxanthin" on October 30, 1998; however, the application emphasizes Purification and preparation of 95% zeaxanthin in microalgae, that is, using a small set of tubes (3 cm in inner diameter and 25 cm in length) to obtain a layered layer, and then a semi-preparative type (inner diameter of 1 cm, length) Purification by 25cm) high pressure liquid chromatography to obtain 95% zeaxanthin, the total recovery rate of zeaxanthin is only 20%, still not ideal, and it still has problems and lacks to be solved.

The inventor is currently engaged in the manufacture and design of related products, and has accumulated many years of practical experience and experience. The invention patent application for the above-mentioned "supercritical fluid anti-solvent microalgae carotenoids and purified zeaxanthin" failed to improve zeaxanthin. The problem of total recovery rate, actively investing in the spirit of innovation and improvement, the preparation of high-content phycocyanin zeaxanthin antioxidant health raw materials.

The technical means to solve the problem of the present invention and the efficacy against the prior art are: different from the above-mentioned invention patent application, the present invention uses a low-pressure column of the leading stage (inner diameter 10 cm, length 30 cm) for partial purification, Purification by high pressure liquid chromatography is required. The purified product of this method contains 40% zeaxanthin, and the total recovery of zeaxanthin is increased by 50%. It has an improvement in efficacy and is the target of its main purpose.

In order to make it easier for those skilled in the art to understand the art to have a deeper understanding of the device content of the present invention and the functional benefits that can be achieved, a specific embodiment will be described and described in detail with reference to the following: a high content of Chlorella The preparation method of the zeaxanthin antioxidant health raw material, wherein: the microalgae used in the invention is marine algae, and the zeaxanthin content per gram of microalgae is higher than 0.9 mg. After removing water by freeze-drying, store at -20 °C for later use.

The modified supercritical carbon dioxide extraction (mSC-CO2) procedure is as follows:

1. Weigh 10g of lyophilized microalgae, put it vertically into the extraction tank, and fill the tank with glass wool to prevent material from seeping out;

2. Preheating the extraction tank and pipeline by oil bath and heating belt; 3. After the temperature reaches 323K, the carbon dioxide is reduced from the cylinder to the 277 K through the refrigeration cycle machine, and the high pressure pump is pressurized at a flow rate of 10 ml/min. Flow into the extraction tank; Fourth, the extraction tank pressure is controlled at 350 bar, the temperature is controlled at 323 K, and the solution ratio is fixed at 60. 5. In the extraction procedure, 15% of the cosolvent ethanol is continuously added by the pump and mixed with carbon dioxide. After uniformization, it enters the extraction tank to extract the algae; sixth, the extract is separated from the carbon dioxide in a separation tank of pressure 50 bar, and the carbon dioxide usage is measured by a wet gas flow meter.

The purification procedure of zeaxanthin is as follows: 1. Take 50g of freeze-dried Chlorella, and add 600mL of dichloromethane to reflux by wire for 20 hours, then concentrate to dryness under reduced pressure to obtain a dry weight of 3.5g. Second, the extract is dissolved back into a mixed solution of ethyl acetate and n-hexane (1:3), and purified by column chromatography. The inner diameter of the column is 10 cm and the length is 30 cm. The column filling material is silica-gel (60-240 mesh); 3. The extraction condition is a mixture of ethyl acetate and n-hexane, the ratio is 1:3 for the extraction, and the zeaxanthin is collected after 72 hours. Draw a layer.

The tendency of scavenging free radicals of DPPH negative ionic compounds and ABTS positive ionic compounds is the same as that of zeaxanthin: when the zeaxanthin concentration is higher, the ability to scavenge DPPH free radicals is better. At the same time, after purification, 40% of the zeaxanthin product was prepared, and a sample of one hundred thousandth (10 ppm) was prepared to prevent the human eye epithelial cells from being inhibited by hydrogen peroxide.

In view of the foregoing, the present invention provides a method for preparing a high-content phycocyanin zeaxanthin antioxidant health raw material, which has been confirmed by the present inventors and after repeated operation tests, and it is confirmed that the functional benefits expected by the present invention can be achieved; It is also the "first creation" that has not yet been seen in the market. It has the "utility value of the industry". It is true that it has already met the founding principle of the invention patent and applied for an invention patent to the bureau in accordance with the provisions of the Patent Law.

Claims (1)

A high-content preparation method of zeaxanthin antioxidant health raw material, wherein the purification procedure of zeaxanthin comprises: Step 1: taking 50g of freeze-dried Chlorella, and adding 600mL of dichloromethane to reflux by wire for 20 hours, Then, the organic layer was concentrated to dryness under reduced pressure to give a dry weight of 3.5 g. Step 2, the extract was then dissolved in a mixed solution of ethyl acetate and n-hexane. The mixture ratio of ethyl acetate and n-hexane was 1 : 3, purified by column chromatography, the inner diameter of the column is 10cm, the length is 30cm, wherein the column filling material is silica-gel (60-240 mesh); step three, the extraction condition is acetic acid A mixture of ethyl ester and n-hexane was eluted in a ratio of 1:3, and a layer containing zeaxanthin was collected after 72 hours.