KR20060085229A - The new method that produce high purity lutein with simultaneously extract and saponification then without using harfull solvent - Google Patents

Abstract

After performing extraction and saponification continuously in marigold, the resulting lutein is a method of producing lutein of about 97% purity using an organic solvent that is harmless to the human body again.

Method of producing new lutein using solvent that is harmless to human body by performing extraction and saponification at the same time

Description

 {The new method that produce high purity lutein with simultaneously extract and saponification then without using harfull solvent}

Carotenoids are yellow, red, and orange pigments that are widely present in nature. Some special carotenoids come with different kinds of fruits

Found in vegetables, bird feathers, egg yolks, shellfish, and human retinal cells. And these carotenoids are found in abundance in marigold, corn and green leafy vegetables.

 Carotenoids absorb light in the visible range of 400-500 nm. The physical properties of these carotenoids are yellow / red. Carotenoids have a paired form that includes isoprene units. Mammals are also forced to ingest these specific carotenoids. Therefore, fruit, vegetables, egg yolks, etc., which are good for carotenoid intake have been known to be very good. In recent years, carotenoids are known to be good for the health and treatment of serious diseases. Carotenoids fall into two classes.

Xanthophyll or hydroxyl-carotenoids, called polar substances, and nonpolar

The substance can be divided into β-carotene, lycopene and the like. But these two

A common feature of classification is that carotenoids contain at least nine double bonds.

This is what determines the color.

Xanthophyll is a hydroxyl or keto group that has a polar function at the end of a double bond.

It has a ring-shaped structure containing it. These substances are mainly lutein,

Zeaxanthin, capsaicin, β-cryptoxanthin, astaxin and the like.

Lutein and zeaxanthin are yellow and orange, respectively.

Lutein may help prevent eye diseases such as macular degeneration and cataracts

It is also generally known that eating carotenoid-rich foods can reduce the incidence of heart disease.

Lutein is present in large quantities in green leafy vegetables such as spinach, kale and broccoli and is found in human blood and in many places in the skin. It is especially found in the retina, iris and optic nerve of the eye.

But mammals can't synthesize these carotenoids, so

Ingestion can only meet the necessary carotenoids.

Lutein and zeaxanthin are present in plants and in ester form.

Generally, carotenoid fatty acid esters are not found in human plasma or blood, and pure forms of lutein, a type of carotenoid, are found in humans. Therefore, it is important to obtain pure lutein economically.

Tyzkowski and hamilton, Poultry Sci., 70 (3): 651-654, 1991

For the first time, a method for the purification of lutein extracted from marigold was introduced.

However, this method is not only time consuming, but also harmful solvents.

Even if used, there was a disadvantage of low yield.

 Many patents have been published on methods for the separation, purification and recrystallization of lutein from marigold (Oleoresin) to obtain pure lutein.

Among them, a patent issued by Khachik in 1995 is to purchase marigold-containing resin extracted from marigold from hexane from kemin, and then saponify to isolate lutein,

It relates to a method for purification and recrystallization.

Patent US, 5,684,564, 1997 relates to a method of obtaining about 70-85% lutein by saponifying marigold containing resin extracted from marigold in boiling hexane at about 70-85 ° C. for 10 hours in propylene glycol and alkaline water.

Disadvantage of this patent is that marigold is extracted from boiling hexane for a long time, so heat-sensitive carotenoids lutein can be denatured or isomerized.

Another disadvantage is that saponification is completed when the saponification is performed at about 70 ° C. for about 10 hours with a solvent such as an aqueous solution of alcohol or propylene glycol having a very low solubility in fatty acid ester lutein. So under heat and air

Oxidation products of lutein may be produced and other side reactions may be carried out.

Therefore, the above invention is not only time-consuming because the saponification and extraction is divided into two stages, there is a disadvantage that the purity of lutein can be obtained only about 70-85%.

 The disadvantage of Khachik's patent, published in 1995, is that it uses a solvent such as dichloromethane, which has the advantage of recrystallization to yield about 97% lutein, but harmful to humans.

Therefore, the technical problem to be achieved by the present invention is easy and economical extraction and separation of lutein esterified with fatty acids and saponification at room temperature

It just wants to complete in a few hours. Therefore, in the existing invention

To overcome the shortcomings and to prepare a lutein of purity of 97% or more to provide a method suitable for human intake, the extraction and purification process of such lutein is a successful and commercially suitable invention.

The carotenoids in marigold are esterified with fatty acids such as lauric, mystic and palmic acid. Khachik in 1995

According to the invention patent marigold has lutein and other main ingredients

It is reported that zeaxanthin, an isomer compound, exists. The present invention

Ethyl acetate, which is harmless to the human body, published by KFDA was selected as the extraction solvent.

Compared with other organic solvents, ethyl acetate is more effective than carotenoids.

Its solubility is high enough to remove these substances from marigold quickly and effectively.

Can be used to extract. Ethyl acetate can also be

Evenly mixed can be used for continuous extraction and saponification.

Since carotenoids are extracted from the homogenized phase, saponification is carried out continuously in a few hours at room temperature. If extracted from a non-uniform solvent such as a mixture of hexane, alcohol and water, saponification is performed.

It is left for a long time at high temperature.

The need for high temperatures to perform saponification can lead to serious problems that can eventually result in the isomerization and denaturation of carotenoids in mass production.

The general method is shown below.

Mix 100 g of fine marigold, 1000 ml of ethyl acetate, 25 g of sodium or potassium hydroxide, and 225 ml of edible ethanol at room temperature.

As it becomes homogeneous, the pH rises to about 11-14, up to 12 in about 2 hours. Edible ethanol may be the most suitable solvent or other used

Solvents include 1-butanol, 2-butanol, 2-methyl-1-propanol, 1-propanol

And 2-propanol may be used. Suitable solvents have a suitable boiling point of about 75 ° C to 120 ° C. Ethanol is suitable because the boiling point is 78 ° C. The pH of the mixture is automatically raised to about 12. The extraction and saponification reaction is completed in about 2 hours. After the reaction, 100 ml of 10% aqueous sodium chloride solution was added and allowed to stand to separate the organic layer and the water layer. Discard the water layer and wash the organic layer three times with hot water. The organic layer is solid

Concentrate until formed, and then cool hexane in 20 ml of ethyl acetate.

Add 30ml to crystallize at room temperature. After filtering the crystals

After washing with ethanol and drying, distilled water in 20 ml of tetrahydrofuran

The addition of 30 ml completes the recrystallization in about 10 minutes.

After separating the crystals and washed three times with cold ethanol and vacuum-dried

About 1.2 g of lutein with 97% purity can be obtained.

 Alternatively, after continuous extraction and saponification, 100 ml of 10% calcium chloride solution is added to separate the organic layer and washed three times with hot water. Then, filtered through a column filled with activated charcoal, concentrated to about 1/5, and then added 100 ml of cold hexane to crystallize at room temperature. The crystals were filtered, washed three times with cold ethanol, and dried in vacuo to obtain approximately 1.2 g of 97% pure lutein. The detailed method is shown in the Example.

 Example 1

    <Continuous Reaction of Extraction and Saponification>

Mix 100 g of fine marigold, 1000 ml of ethyl acetate and 250 ml of 10% ethanol potassium chloride (made by adding 25 g of potassium chloride to 225 ml of ethanol) at room temperature. For continuous reactions of extraction and saponification, the pH must be above 12. Therefore, always monitor with automatic pH meter.

In order to raise the pH above 12,

10% ethanol potassium chloride is required. However, if an excessive amount of base is used in the extraction and saponification process, it is important to use an appropriate amount of base since it may consume too much water as well as time in later purification.

The extraction and saponification process can be examined by thin layer chromatography (detection of nucleic acid / acetone = 90/10 as a developing solvent on n-silica gel thin film) and high performance liquid chromatography (Khachik J, Chrom 449: 119-133, 1998). have.

After 2 hours the carotenoid esters have completed saponification.

After the reaction, the mixture was placed in a separatory funnel, and then 100 ml of 10% sodium chloride solution was added and allowed to stand. After a while the layer is divided into two. Discard the lower layer and wash the upper layer about 3 times with 1000 ml of hot water. After cleaning

The organic layer is concentrated and washed until a solid is formed. The resulting solid was dissolved in 20 ml of ethyl acetate, and 30 ml of cold hexane was added to crystallize at room temperature. The crystals were filtered off, washed with 100 ml of cold ethanol and dried in a high vacuum for one day, yielding about 2.0 g (70%) lutein.

The components of lutein obtained here are shown in Table 1.

Carotenoid Components of 70% Lutein Isolated from Marigold Extract Carotenoids Component ratio (%) all-E-Lutein 93.0 Total-Z-Lutein 0.3 all-E-Zeaxanthin 5.4 Anhydroluteins 0.8 alpha-Cryptoxanthin 0.2 beta-Cryptoxanthin 0.3

                          Table 1.

    Purification through recrystallization

Recrystallization is started by dissolving lutein crystals with a purity of 70% 2.0 g in 20 ml of tetrahydrofuran and adding 30 ml of distilled water. Leave at room temperature for about 10 minutes and centrifuge to separate solids. Lutein after removing the solution

The crystals were washed with 20ml ethanol and then centrifuged again and dried for one day at high vacuum and room temperature to obtain about 97% high purity lutein.

have.

Filtration through filtration

 After continuous extraction and saponification, 100 ml of 10% aqueous sodium chloride solution is added to separate the water and organic layers. The organic layer was washed three times with hot water and filtered through a column filled with activated carbon.

Concentrate the filtrate to about 1/5 and add 100 ml of cold hexane to crystallize at room temperature. The crystals were filtered and washed three times with cold ethanol and vacuum dried to obtain approximately 1.2 g of 97% pure lutein.

The present invention discloses a method for producing economically pure lutein using a solvent that is harmless to the human body as a patent for separation and purification of lutein. Therefore, lutein prepared by the present invention is a nutritional supplement or

It can be used as a food additive.

Claims (1)

1. Extract marigolds using ethyl acetate solvent and alcohol solvent. New method of obtaining lutein by performing saponification successively. 2. A new way to prevent lutein oxidation and denaturation by completing extraction and saponification in 2 hours at room temperature. 3. Monitor pH during continuous extraction and saponification, Further adjusting the pH by adding a base. 4. A method wherein successive extraction and saponification are carried out in the presence of a base. 5. Dissolve the lutein crystals in tetrahydrofuran and use activated carbon New way to filter. 6. A new way to recrystallize using solvents that are harmless to the human body. 7. A new method of recrystallization in a solution containing tetrahydrofuran. 8. Economical and effective way to obtain high purity lutein of more than 97%.