WO2020073976A1

WO2020073976A1 - Industrial preparation method of lutein and marigold flavone

Info

Publication number: WO2020073976A1
Application number: PCT/CN2019/110505
Authority: WO
Inventors: 连运河; 吴迪; 高伟; 安晓东; 王欢欢
Original assignee: 晨光生物科技集团股份有限公司
Priority date: 2018-10-12
Filing date: 2019-10-11
Publication date: 2020-04-16
Also published as: CN109232346A; MX2021004224A; CN109232346B

Abstract

The present invention relates to an industrial preparation method of lutein and marigold flavone. Main improvement points thereof are: mixing a petroleum ether and a flavone extraction solvent; extracting marigold particles; adding a strong-polarity solvent to the flavone extraction solvent after extraction is finished; and further promoting separation of the lutein and the marigold flavone. The present invention uses a double-solvent extraction method, so that the extraction efficiency is improve; the purity of the marigold flavone is improved; furthermore, the method in the present invention is simple in steps and low in costs, and is applicable to industrial large-scale production; the purity of the extracted marigold flavone is high; and the marigold flavone has various bioactivities and has a wide application prospect.

Description

Industrial preparation method of lutein and marigold flavonoids

cross reference

This application requires the priority of the Chinese patent application No. 201811188771.2 with the patent title of "an industrial preparation method of lutein and marigold flavonoids" filed on October 12, 2018, the entire disclosure of which is incorporated by reference in its entirety.

Technical field

The invention relates to the field of extracting natural products, in particular to a method for extracting lutein and flavonoids from marigold flower granules.

Background technique

Marigold is one of the main cultivated potted plants in China, and it is widely used in indoor and outdoor environment layout. With the development of nutrition and health industry, the international demand for marigold is increasing. Marigold not only has ornamental value, but also has very good medicinal value. Its main pharmacological components are flavonoids, carotenoids, terpenes, glycosides, essential oils and other components.

Marigold flower is the main source of lutein. After extracting lutein, a large amount of marigold residue is discarded as industrial waste or used as feed. This has a certain impact on the environment and also causes a lot of waste of resources. Research reports that marigold flowers are rich in flavonoids, such as quercetin marigold, etc., accounting for about 2% to 11% of marigold flowers. Studies have shown that these flavonoids have good biological activities, such as antioxidant, anti-inflammatory, anti-allergic, blood pressure lowering, anti-arrhythmic, anti-tumor effects. At the same time, there are some other active ingredients in marigold flowers, such as oligosaccharides, glycosides and other compounds to be developed.

ZL201410104645 discloses a method for extracting and separating lutein and flavonoids in marigold. The method is to add marigold petal powder to the isopropyl alcohol-potassium hydroxide two-phase system for extraction, and the extraction rate of lutein reaches 86.73%. The extraction rate of flavonoids reached 78.31%. The patent believes that it overcomes the shortcomings of the large amount of organic solvents used in production, high material consumption and high energy consumption. However, its material ratio is from 1 to 400 to 1 to 2000, which requires a large amount of extraction solvent, which increases the cost, and at the same time causes a large amount of industrial wastewater, which causes a burden on the environment and is difficult to achieve industrialization. At the same time, the extraction rate of lutein by this patent method is low, only 86.73%.

Summary of the invention

The object of the present invention is to provide a method for simultaneously extracting lutein and marigold flavonoids, and the extraction is achieved by mixing solvents, which specifically includes the following steps:

1) Mix petroleum ether and flavonoid extraction solvent to extract marigold flower particles to obtain a mixed extract;

2) Add a strong polar solvent with a polarity greater than that of the flavonoid extraction solvent to the mixed extract, shake it well, and leave it to separate into layers to obtain a lutein phase and a flavonoid phase. The petroleum ether or flavonoid extraction solvent is removed to obtain marigold flavonoid and lutein extract.

In the present invention, a mixture of petroleum ether and flavonoid extractant is used to extract marigold flowers particles. During the extraction of the raw materials of the above two organic phases, lutein and flavonoids are quickly distributed into the corresponding organic phases, compared to the single substance extraction The purity and efficiency are high, and the azeotropic point of the two solvents after mixing is lower than that of a single solvent, the overall consumption of the solvent during the extraction process is less, and the temperature during the extraction process is lower. Further, after the extraction is completed, adding a strong polar solvent with a polarity greater than the extractant of the flavonoids to the flavonoid extraction phase can promote better distribution of flavonoids and lutein in the two phases, and improve the yield and purity of the two products .

Preferably, the flavonoid extraction solvent is any one, a mixture of two or three of methanol solution, acetone solution, ethyl acetate, n-butanol and ethanol solution.

Further preferably, the flavonoid extraction solvent is a methanol solution or an acetone solution.

Preferably, when the flavonoid extraction solvent is a methanol solution, the methanol volume fraction is 60-100%.

More preferably, the volume fraction of methanol is 80-100%

Preferably, the strong polar solvent is any one or a mixed solvent of ethylene glycol, methanol solution, ethanol solution or water.

Further preferably, the strong polar solvent is water.

When the flavonoid extraction solvent is a methanol solvent, if the methanol or ethanol solution solvent is selected as the strong polar solvent, to ensure greater polarity, the volume fraction of the methanol and ethanol solution in the strong polar solvent is less than the flavonoid The volume fraction of methanol in the extraction solvent.

Preferably, the addition amount of the strong polar solvent is 0.2 to 4 times the volume of the flavonoid extraction solvent.

As a further preferred solution, the extraction solvent of the flavonoid is a methanol solution with a volume fraction of 80-100%, the strong polar solvent is water, and the volume ratio of the methanol solution to water is 1: 0.2-1; more preferably , The volume ratio of the methanol solution to water is 1: 0.2 ～ 0.4

Or, the extraction solvent of the flavonoid is acetone, the highly polar solvent is water, and the volume ratio of acetone to water is 1: 0.2-2; more preferably, the volume ratio of acetone to water is 1: 0.2 ～ 1.5.

The methanol solution with a volume fraction of 80-100% is slightly higher than acetone as the extraction solvent. However, due to the strong polarity of methanol, the overall structure of the marigold flower particles will be destroyed after extraction, but the acetone extraction does not exist. Therefore, in actual production, a suitable solvent can be selected according to the industrial extraction method. For example, when extraction is carried out by immersion, methanol solution can be used as the extraction agent. When extraction is carried out by leaching, the extraction efficiency can be avoided to avoid particle damage. Reduce, you can choose acetone as an extractant.

As a more preferred solution, the extraction solvent of the flavonoid is a methanol solution with a volume fraction of 80-100%, the strong polar solvent is water, and the volume ratio of the methanol solution to water is 1: 0.2-0.4.

The volume ratio of the petroleum ether to the methanol solution is 1: 0.8 to 1.2, and the ratio of the total volume of the petroleum ether and methanol solution to the volume-mass of the marigold flower particles is 4 to 6: 1.

Or, the extraction solvent of the flavonoid is acetone, the highly polar solvent is water, the volume ratio of the acetone to water is 1: 0.2 to 1.5, and the volume ratio of the petroleum ether to the acetone is 1: 0.8 ~ 1.2, the ratio of the total volume of the petroleum ether and the acetone to the volume-mass of the marigold flower particles is 4-6: 1.

The above scheme using acetone as an extractant has a better effect when the volume ratio of acetone to water is 1: 1.3 to 1.5.

Under the above scheme, the yield of lutein is greater than 99%, and the yield and purity of flavonoid extract are also high, which is a preferred mode of operation.

Preferably, the volume ratio of the petroleum ether to the flavonoid extraction solvent is 1: 0.2 to 3, more preferably 0.4 to 1. The relative amount of petroleum ether and flavonoid extractant will affect the distribution of the extracted lutein and flavonoids in the above two solvents. If the amount of one of the solvents is too much, the active ingredients extracted by the other phase will be more dissolved into Among them, therefore, on the premise of ensuring that both solvents can fully extract lutein and flavonoids, the relative amounts of the above-mentioned solvents are beneficial to improve the purity of the two products.

Further preferably, the flavonoid extractant is acetone, and the volume ratio of petroleum ether to acetone is 1: 0.4 to 1.

Further preferably, the flavonoid extractant is a methanol solution, and the volume ratio of petroleum ether to methanol solution is 1: 0.8 to 1.

Preferably, the ratio of the total volume of the petroleum ether and flavonoid extraction solvent to the volume and mass of the marigold flower particles is 2-8: 1. Under the above dosage, full extraction of lutein and flavonoids in marigold flower granules can be achieved

Preferably, the extraction temperature is 30-55 ° C. Choosing the extractant of the present invention to mix with petroleum ether can effectively reduce the azeotropic point of the mixed extractant and reduce the extraction temperature, and sufficient extraction can be performed at the above temperature.

Preferably, the extraction conditions of the present invention are extraction by countercurrent extraction, the extraction time is 0.5 to 4 hours, and the extraction temperature is 30 to 55 ° C.

Preferably, the present invention also includes the operation of refining the flavonoids, including the following steps: adding a mixture of water and ethyl acetate to the marigold flavonoids, shaking repeatedly, leaving to stand at room temperature, collecting the ethyl acetate phase, and drying the acetic acid Ethyl acetate, to obtain refined marigold flavonoids;

Further preferably, the volume ratio of water to ethyl acetate is 1: 1-10;

Further preferably, the mass-volume ratio of the mixed liquid of marigold flavonoids and the water and ethyl acetate is 1: 1 to 3.

Another object of the present invention is to protect the flavonoids of marigold extracted by the method of the present invention.

The last object of the present invention is to protect the lutein extract prepared by the method of the present invention.

The invention has the following beneficial effects:

1) The present invention uses a mixed solution of petroleum ether and flavonoid extractant to extract marigold flowers particles. During the extraction of the dual organic phase, lutein and flavonoids are quickly distributed into the corresponding organic phase, which is more efficient than a single solvent for extracting a single substance high. In addition, the present invention finds that, after the extraction of the flavonoids, the addition of a strong polar solvent more polar than the extractant in the organic phase can further promote the separation of lutein and flavonoids and improve the extraction effect.

2) If you want to achieve the same extraction effect, the dual solvent selected by the present invention has a lower extraction temperature than a single solvent, and the amount of extraction solvent is small, which can greatly save energy consumption, be more environmentally friendly, and more suitable for industrial production and application. .

3) The present invention further proposes a preferred extraction solvent combination, which can effectively improve the extraction effect.

4) The present invention simultaneously extracts lutein and marigold flavonoids. Since marigold flavonoids have a good antioxidant effect, the oxidation loss of lutein is protected to a certain extent.

detailed description

The following examples are used to illustrate the present invention, but are not used to limit the scope of the present invention.

The marigold flower particles involved in the examples are marigold flower particles obtained from marigold flowers through lactic acid fermentation, pressing, drying, crushing, and granulation.

The specific operation of the extraction involved in the examples is immersion extraction. Except for the operating conditions given in the examples and comparative examples, the other operating conditions are identical.

Example 1

This embodiment relates to the extraction method of the present invention, which includes the following steps:

1) Add 50L of petroleum ether and 125L of 90% methanol solution to 50kg of marigold flower pellets, immerse and extract at 50 ° C for 1h, repeat extraction 4 times according to the above conditions, and combine the extracts.

2) Add 40L of purified water to the extract, shake and let stand for layering. The petroleum ether phase is concentrated to obtain lutein extract, and the methanol phase is concentrated and dried to obtain marigold flavonoids.

3) Add 10L of a mixed solvent of water and ethyl acetate (1: 5) to the 3kg crude marigold flavonoid extract, shake repeatedly, let stand at room temperature, collect the ethyl acetate phase, and dry the ethyl acetate to obtain marigold flavonoids .

Example 2

1) To 50 kg of marigold flower particles, 125 L of petroleum ether and 125 L of acetone are added, and extraction is carried out at 50 ° C. for 1 h, and extraction is repeated 4 times according to the above conditions, and the extracts are combined.

2) Add 175L of purified water to the extract, shake and let stand for layering. The petroleum ether phase is concentrated to obtain lutein extract, and the acetone phase is concentrated and dried to obtain marigold flavonoids.

Example 3

1) To 50 kg of marigold flower pellets, 125 L of petroleum ether and 125 L of ethyl acetate were added, and extraction was carried out at 50 ° C. for 1 h, and extraction was repeated 4 times under the above conditions, and the extracts were combined.

2) Add 175 L of 50% ethanol solution to the extract, shake and let stand for layering. The petroleum ether phase is concentrated to obtain lutein extract, and the ethyl acetate phase is concentrated and dried to obtain marigold flavonoids.

Example 4

2) Add 125L of 50% methanol to the extract, shake and let stand for layering. The petroleum ether phase is concentrated to obtain lutein extract, and the acetone methanol solution is concentrated and dried to obtain marigold flavonoids.

Example 5

2) Add 25L of aqueous solution to the extract, shake and let stand for layering. The petroleum ether phase is concentrated to obtain lutein extract, and the acetone phase is concentrated and dried to obtain marigold flavonoids.

Example 6

1) To 50 kg of marigold flower particles, 125 L of petroleum ether and 50 L of acetone are added, and extraction is carried out at 50 ° C. for 1 h, and extraction is repeated 4 times according to the above conditions, and the extracts are combined.

2) Add 70L of aqueous solution to the extract, shake and let stand for layering. The petroleum ether phase is concentrated to obtain lutein extract, and the acetone phase is concentrated and dried to obtain marigold flavonoids.

Comparative Example 1

Compared with Example 1, the difference between this comparative example is that in step 1), lutein is not extracted with petroleum ether, and only the step of extracting methanol solution is involved, and the conditions for extracting marigold particles with methanol solution are: Add 50L of 90% methanol solution to 50kg of marigold flower pellets, extract at 50 ℃ for 1h, and repeat extraction 4 times according to the above conditions. The extracts were combined and concentrated to dry to obtain marigold flavonoids.

Comparative Example 2

Compared with Example 1, the difference between this comparative example is that in step 1), simultaneous extraction is not carried out. First, petroleum ether is used to extract lutein, and then methanol solution is used to extract flavonoids, including the following steps:

1) Petroleum ether added to the marigold flower particles is extracted at 50 ° C for 1 hour, repeatedly extracted 4 times, and the extract is concentrated to obtain lutein extract;

2) Dry the marigold flower meal after extracting lutein, take 50 kg of marigold flower meal after drying, add 125L of 90% methanol solution to it, and extract at 50 ℃ for 1h, repeat according to the above conditions Extract 4 times. The extracts were combined and concentrated to dry to obtain marigold flavonoids.

Comparative Example 3

1) Petroleum ether added to the marigold flower pellets is extracted at 50 ° C for 1 hour, repeatedly extracted 4 times, and the extract is concentrated to obtain lutein extract;

2) Dry the marigold flower meal after extracting lutein, take 50kg of the dried marigold flower meal, add 350L of 90% methanol solution to it, extract at 60 ℃ for 1h, and repeat the extraction according to the above conditions 6 Times. The extracts were combined and concentrated to dry to obtain marigold flavonoids.

Comparative Example 4

Compared with Example 1, the difference is that after the extraction is completed, no strong polar solvent is added to the flavonoid extraction phase. The specific implementation steps are as follows: adding 50L of petroleum ether and 125L of 90% methanol solution to 50kg of marigold flower pellets, extracting at 50 ° C for 1h, repeating the extraction 4 times according to the above conditions, combining the extracts, and standing for separation . The petroleum ether phase is concentrated to obtain lutein extract, and the methanol phase is concentrated and dried to obtain marigold flavonoids.

Experimental example

The products extracted in the examples and comparative examples were tested for lutein by ultraviolet detection method and flavonoids by high-performance liquid method. The results are shown in the following table:

It can be seen from Comparative Example 1 that the extraction of marigold flower particles is only extracted with methanol, and the extraction effect is inferior to that of mixed solvent extraction. It can be seen from Comparative Example 2 that even after the extraction of lutein, the material is further extracted with the same flavonoid extraction solvent under the same conditions, and the extraction effect is not as good as simultaneous extraction under the same conditions. It can be seen from Comparative Example 3 that after extracting lutein, the material is further extracted with the same flavonoid extraction solvent. To achieve the same extraction effect as simultaneous extraction, it is necessary to increase the amount of extractant, increase the extraction temperature, and perform more Repeated extraction for many times, which further proves that the mixed solvent extraction is more efficient, more energy-saving and more suitable for industrial production. It can be known from Comparative Example 4 that after the extraction is completed or no strong polar solvent is added, the extraction effect of lutein and flavonoids will obviously deteriorate.

Although the present invention has been described in detail with general description, specific embodiments and tests, but based on the present invention, some modifications or improvements can be made to it, which is obvious to those skilled in the art . Therefore, these modifications or improvements made on the basis of not deviating from the spirit of the present invention belong to the scope claimed by the present invention.

Industrial applicability

The invention provides an industrial preparation method of lutein and marigold flavonoids. The invention mixes petroleum ether and flavonoid extraction solvent to extract marigold flower particles. After the extraction is completed, a strong polar solvent is added to the flavonoid extraction solvent to further promote the separation of lutein and marigold flavonoids. The invention adopts the double solvent extraction method, which can improve the extraction efficiency and the purity of marigold flavonoids. Moreover, the method of the invention has simple steps and low cost, and is suitable for industrial large-scale production. The extracted marigold flavonoids have high purity and many This kind of biological activity has good economic value and application prospects.

Claims

The industrial preparation method of lutein and marigold flavonoids is characterized by the following steps:

1) Mix petroleum ether and flavonoid extraction solvent to extract marigold flower particles to obtain a mixed extract;

2) Add a strong polar solvent with a polarity greater than that of the flavonoid extraction solvent to the mixed extract, shake it well, and leave it to separate into layers to obtain a lutein phase and a flavonoid phase. Separate the two phases to remove petroleum ether or flavonoids, respectively Extract the solvent to obtain marigold flavonoids and lutein extract.
The method according to claim 1, wherein the flavonoid extraction solvent is any one, a mixture of two or three of methanol solution, acetone solution, ethyl acetate, n-butanol and ethanol solution, preferably Methanol solution and acetone; further preferably, the volume fraction of methanol in the methanol solution is 60-100%.
The method according to claim 1 or 2, wherein the strongly polar solvent is any one or a mixed solvent of ethylene glycol, methanol solution, ethanol solution or water, preferably water.
The method according to any one of claims 1 to 3, wherein the addition amount of the strong polar solvent is 0.2 to 4 times the volume of the flavonoid extraction solvent.
The method according to any one of claims 1 to 4, wherein the flavonoid extraction solvent is a methanol solution with a volume fraction of 80 to 100%, the strong polar solvent is water, and the methanol solution and the The volume ratio of water is 1: 0.2 ～ 1;

Or, the extraction solvent of the flavonoid is acetone, the highly polar solvent is water, and the volume ratio of the acetone to water is 1: 0.2-2.
The method according to any one of claims 1 to 5, wherein the volume ratio of the petroleum ether to the flavonoid extraction solvent is 1: 0.2 to 3, preferably 0.4 to 1.
The method according to any one of claims 1 to 6, wherein the ratio of the total volume of the petroleum ether and flavonoid extraction solvent to the volume and mass of the marigold flower particles is 2 to 8: 1; and / or, The extraction temperature is 30 to 55 ° C.
The method according to any one of claims 1 to 7, further comprising an operation of purifying the flavonoid extract;

Preferably, a mixed solution of water and ethyl acetate is added to the marigold flavones, repeatedly shaken, and left to stand, the ethyl acetate phase is collected, and concentrated to dry to obtain purified marigold flavonoids.
Marigold flavonoids extracted by the method according to any one of claims 1 to 8.
The lutein extract extracted by the method according to any one of claims 1 to 8.