Method for extracting lutein from marigold and application of lutein

Abstract

The invention belongs to the technical field of extraction of functional effective components from natural plants, and particularly relates to a method for extracting xanthophyll from marigold and application of the xanthophyll. The extraction method mainly comprises the following steps: granulating, infiltrating, extracting, concentrating and washing. According to the method, marigold particles of different specifications are mixed according to a certain proportion, then the marigold particles are infiltrated with the organic solvent, then the chlorogenic acid, betaine and histidine mixed solution with the pH being 8-10 is added for auxiliary extraction, and then the xanthophyll is obtained through concentration, washing and drying, so that the extraction rate of the xanthophyll in the marigold is increased, and the yield of the xanthophyll in the marigold is increased. The extracted lutein can be applied to animal feed.

Description

Technical Field

[0001] The present invention belongs to the technical field of extracting functional active ingredients from natural plants, and particularly relates to a method for extracting lutein from marigold and its application. Background Art

[0002] Lutein mainly exists in vegetables and fruits in nature, especially in dark green leafy vegetables and yellow-orange fruits such as marigold, cauliflower, pumpkin, spinach, parsley, etc. Among them, marigold has the highest lutein content and is the main raw material for extracting lutein industrially at present.

[0003] In the prior art, lutein is extracted from marigold by traditional organic solvent extraction methods, such as adding single or mixed solvents such as acetone, n-hexane, petroleum ether, dichloromethane, chloroform, tetrahydrofuran, ethanol, etc. After extraction, filtration, concentration, and removal of solvent residues, lutein resin or lutein extract with lutein ester as the main component is obtained. Then, an alcoholic solution of inorganic base is used as a saponification solution for saponification reaction, and lutein crystals are obtained after purification. This process of separately performing extraction and saponification as two steps has problems such as complex operation, long time consumption, and large amounts of organic solvents and inorganic bases used. Summary of the Invention

[0004] In order to overcome the above deficiencies of the prior art, the present invention provides a method for extracting lutein, a preparation thereof, and its application.

[0005] In order to achieve the above object, the present invention adopts the following technical solutions: A method for extracting lutein from marigold, comprising the following steps: S1 Granulation: The dried marigold flowers are crushed into powder, and then made into small particles of different specifications of 0.1 - 1.0 mm, 2 - 3 mm, and 3 - 5 mm through a granulator. The particles of different specifications are mixed to obtain marigold flower mixed particles.

[0006] Further, the mass ratio of the 0.1 - 1.0 mm, 2 - 3 mm, and 3 - 5 mm particles is 1:(1 - 3):1 - 2.

[0007] S2 Infiltration: One or more solvents in short-chain alkanes or their derivatives are added to the marigold flower mixed particles for ultrasonic infiltration to obtain marigold infiltration liquid.

[0008] Preferably, the number of carbon atoms of the short-chain alkane in S2 ≤ 6, and the derivative of the alkane is an alcohol, acid, or ester of the alkane. The ratio of the volume of the solvent to the mass of the marigold mixture particles is 5 - 10:1.

[0009] More preferably, the infiltration solvent is ethanol and ethyl acetate, and the mixing volume ratio is 2 - 3:1.

[0010] S3 Extraction: Add a mixed solution of chlorogenic acid, betaine, and histidine to the marigold infusion for extraction, and filter to obtain the marigold extract.

[0011] Preferably, the mass ratio of chlorogenic acid, betaine, and histidine in S3 is 1 - 2:1:3.

[0012] Preferably, the mass fraction of the mixed solution of chlorogenic acid, betaine, and histidine is 10 - 20%.

[0013] Preferably, the pH of the mixed solution of chlorogenic acid, betaine, and histidine is 8 - 9, and the pH regulator used is sodium hydroxide or potassium hydroxide.

[0014] Preferably, the content of chlorogenic acid, betaine, and histidine in the marigold extract in S3 is 10 - 20%.

[0015] Preferably, the extraction time in S3 is 1 - 6 hours.

[0016] S4 Concentration: Subject the marigold lutein extract to vacuum low-temperature distillation to remove organic solvents and water, collect the concentrated solution, and obtain the crude lutein extract.

[0017] Preferably, the vacuum low-temperature distillation conditions in S4 are: 50°C ≤ temperature ≤ 60°C.

[0018] S5 Washing: Wash the crude lutein extract with deionized water, and freeze-dry the insoluble matter after washing at -40°C to obtain lutein.

[0019] Preferably, the mass ratio of the crude lutein extract to water is 1:10 - 20.

[0020] The present invention also provides the application of the above marigold lutein in animal feed.

[0021] When the marigold lutein of the present invention is applied to animal feed, it can increase the egg production of poultry and also improve the color of aquatic animals. Beneficial Effects

[0022] After mixing different specifications of marigold particles in a certain proportion in the present invention, the infiltration efficiency of organic solvents is improved, and thus the extraction rate of marigold lutein is increased.

[0023] By adding chlorogenic acid, betaine, and histidine to the infiltration solution for auxiliary extraction in the present invention, the stability of lutein in the marigold extraction process is improved, and thus the purity of lutein in the crude lutein extract is increased.

[0024] In step S4 of the present invention, organic solvents ethanol and ethyl acetate are removed by vacuum distillation, so that water-insoluble lutein dissolves out to form insoluble substances, while chlorogenic acid, betaine and histidine are easily soluble in water, thereby realizing the effective separation of lutein from chlorogenic acid, betaine and histidine, and improving the yield and purity of lutein; at the same time, by means of water washing, the purity of lutein is further improved. Specific embodiments

[0025] The technical solutions of the present invention are further described below in conjunction with specific embodiments. The raw materials used in the following embodiments can be obtained from conventional commercial channels without special instructions; the processes used, without special instructions, are all conventional processes in the art.

[0026] Example 1 A method for extracting lutein from marigold, comprising the following steps: S1 Granulation: The dried marigold flowers are crushed into powder, and then made into fine particles of different specifications of 0.1-1.0 mm, 2-3 mm, and 3-5 mm through a granulator. The 0.1-1.0 mm, 2-3 mm, and 3-5 mm are in a mass ratio of 1:3:2 to obtain a marigold flower mixed granule.

[0027] S2 Infiltration: Add 5 times the volume of a mixed solvent to 1 part by mass of the marigold flower mixed granule. The volume ratio of ethanol to ethyl acetate in the mixed solvent is 2:1, and ultrasonic treatment is carried out at 60 °C for 1 h with 800 W to obtain a marigold infiltration solution.

[0028] S3 Extraction: Add 1 part by mass of a mixed solution of chlorogenic acid, betaine and histidine (the total mass fraction of the solute in the mixed solution is 10%, the solute is composed of chlorogenic acid, betaine and histidine in a mass ratio of 2:1:3, and the solvent is water) to 10 parts by mass of the marigold infiltration solution. The pH of the mixed solution is adjusted to 9 with sodium hydroxide, and then stirring extraction is carried out at 40 °C and 250 rpm for 2 h to obtain a marigold extraction solution.

[0029] S4 Concentration: The marigold lutein extraction solution is subjected to vacuum distillation at a pressure of 0.015 MPa at 60 °C until ethanol is undetectable, and the insoluble substances in the remaining distillation materials are collected to obtain a crude lutein extract; S5 Washing: The crude lutein extract in S4 is washed with 20 times its mass of deionized water, and the washed insoluble substances are freeze-dried at -40 °C until the water content is 0.17 wt% to obtain the lutein.

[0030] Example 2 A method for extracting lutein from marigold, comprising the following steps: S1 Granulation: The dried marigold flowers are crushed into powder, and then made into small particles of different specifications of 0.1 - 1.0 mm, 2 - 3 mm, and 3 - 5 mm through a granulator. The 0.1 - 1.0 mm, 2 - 3 mm, and 3 - 5 mm particles are mixed according to a mass ratio of 1:1:1 to obtain marigold flower mixed particles.

[0031] S2 Infiltration: Add 10 times the volume of ethanol solvent to 1 part by mass of marigold flower mixed particles, and perform ultrasonic treatment at 800 W for 1 h at 60 °C to obtain marigold infiltration liquid.

[0032] S3 Extraction: Add 1 part by mass of a mixed solution of chlorogenic acid, betaine, and histidine (the mass fraction of the mixed solution is 10%, and the mass ratio of chlorogenic acid, betaine, and histidine is 1:1:3, and the solvent is water) to 5 parts by mass of marigold infiltration liquid. Adjust the pH of the mixed solution to 9 with sodium hydroxide, and then perform stirring extraction at 40 °C and 250 rpm for 2 h to obtain marigold extract.

[0033] S4 Concentration: Carry out vacuum distillation of the marigold lutein extract at 50 °C under a pressure of 0.015 MPa until ethanol is no longer detected. Collect the insoluble matter in the remaining material after distillation to obtain crude lutein extract. S5 Washing: Wash the crude lutein extract in S4 with 15 times its mass of deionized water, and freeze-dry the washed insoluble matter at -40 °C until the water content is 0.15 wt% to obtain the said lutein.

[0034] Example 3 A method for extracting lutein from marigold flowers, comprising the following steps: S1 Granulation: The dried marigold flowers are crushed into powder, and then made into small particles of different specifications of 0.1 - 1.0 mm, 2 - 3 mm, and 3 - 5 mm through a granulator. The 0.1 - 1.0 mm, 2 - 3 mm, and 3 - 5 mm particles are mixed according to a mass ratio of 1:2:2 to obtain marigold flower mixed particles.

[0035] S2 Infiltration: Add 8 times the volume of ethyl acetate solvent to 1 part by mass of marigold flower mixed particles, and perform ultrasonic treatment at 800 W for 1 h at 60 °C to obtain marigold infiltration liquid.

[0036] S3 Extraction: Add 1.5 parts by mass of a mixed solution of chlorogenic acid, betaine, and histidine (the mass fraction of the mixed solution is 20%, and the mass ratio of chlorogenic acid, betaine, and histidine is 2:1:3, and the solvent is water) to 10 parts of marigold infiltration liquid. Adjust the pH of the mixed solution to 9 with sodium hydroxide, and then perform stirring extraction at 40 °C and 250 rpm for 2 h to obtain marigold extract.

[0037] S4 Concentration: Carry out vacuum distillation on the marigold lutein extract at 60°C under a pressure of 0.015 MPa until ethanol is no longer detectable, collect the insoluble matter in the remaining distillation material, and obtain the crude lutein extract; S5 Washing: Wash the crude lutein extract in S4 with 10 times its mass of deionized water, and freeze-dry the washed insoluble matter at -40°C until the water content is 0.18 wt%, to obtain the lutein.

[0038] Comparative Example 1 A method for extracting lutein from marigold, comprising the following steps: S1 Granulation: Crush the dried marigold flowers into powder, and then use a granulator to make fine particles with a specification of 0.1 - 1.0 mm, to obtain marigold flower particles.

[0039] S2 Infiltration: Add 5 times the volume of a mixed solvent to 1 part by mass of the marigold flower mixed particles. The volume ratio of ethanol to ethyl acetate in the mixed solvent is 2:1, and perform ultrasonic treatment at 800 W for 1 h at 60°C to obtain the marigold infiltration liquid.

[0040] S3 Extraction: Add 1 part by mass of a mixed solution of chlorogenic acid, betaine, and histidine (the mass fraction of the mixed solution is 10%, and the mass ratio of chlorogenic acid, betaine, and histidine is 2:1:3, and the solvent is water) to 10 parts by mass of the marigold infiltration liquid. Adjust the pH of the mixed solution to 9 with sodium hydroxide, and then carry out stirring extraction at 40°C and 250 rpm for 2 h to obtain the marigold extract.

[0041] S4 Concentration: Carry out vacuum distillation on the marigold lutein extract at 60°C under a pressure of 0.015 MPa until ethanol is no longer detectable, collect the insoluble matter in the remaining distillation material, and obtain the crude lutein extract; S5 Washing: Wash the crude lutein extract in S4 with 10 times its mass of deionized water, and freeze-dry the washed insoluble matter at -40°C until the water content is 0.16 wt%, to obtain the lutein.

[0042] Comparative Example 2 A method for extracting lutein from marigold, comprising the following steps: S1 Granulation: Crush the dried marigold flowers into powder, and then use a granulator to make fine particles with different specifications of 0.1 - 1.0 mm, 2 - 3 mm, and 3 - 5 mm. Mix 0.1 - 1.0 mm, 2 - 3 mm, and 3 - 5 mm according to a mass ratio of 1:3:2 to obtain the marigold flower mixed particles.

[0043] S2 Infiltration: Add 5 times the volume of a mixed solvent to 1 part by mass of the marigold flower mixed particles. The volume ratio of ethanol to ethyl acetate in the mixed solvent is 2:1, and perform ultrasonic treatment at 800 W for 1 h at 60°C to obtain the marigold infiltration liquid.

[0044] S3 Extraction: Filter the infiltration liquid to obtain the marigold extract.

[0045] S4 Concentration: Carry out vacuum distillation on the marigold lutein extract at 60 °C under a pressure of 0.015 MPa until ethanol cannot be detected, collect the insoluble matter in the remaining distillation materials, and obtain the crude lutein extract. S5 Washing: Wash the crude lutein extract in S4 with deionized water 10 times its mass, and freeze-dry the washed insoluble matter at -40 °C until the water content is 0.17 wt% to obtain the lutein.

[0046] Comparative Example 3 A method for extracting lutein from marigold, comprising the following steps: S1 Granulation: Crush the dried marigold flowers into powder, and then use a granulator to make fine particles of different specifications of 0.1 - 1.0 mm, 2 - 3 mm, and 3 - 5 mm. Mix 0.1 - 1.0 mm, 2 - 3 mm, and 3 - 5 mm according to a mass ratio of 1:3:2 to obtain the marigold flower mixed particles.

[0047] S2 Infiltration: Add 5 times the volume of the mixed solvent to 1 mass portion of the marigold flower mixed particles. The volume ratio of ethanol to ethyl acetate in the mixed solvent is 2:1, and perform ultrasonic treatment at 800 W for 1 h at 60 °C to obtain the marigold infiltration liquid.

[0048] S3 Extraction: Add 1 mass portion of the betaine and histidine mixed solution (the mass fraction of the mixed solution is 10%, the mass ratio of betaine to histidine is 1:3, and the solvent is water) to 10 mass portions of the marigold infiltration liquid. Adjust the pH of the mixed solution to 9 with sodium hydroxide, and then carry out stirring extraction at 40 °C and 250 rpm for 2 h to obtain the marigold extract.

[0049] S4 Concentration: Carry out vacuum distillation on the marigold lutein extract at 60 °C under a pressure of 0.015 MPa until ethanol cannot be detected, collect the insoluble matter in the remaining distillation materials, and obtain the crude lutein extract. S5 Washing: Wash the crude lutein extract in S4 with deionized water 10 times its mass, and freeze-dry the washed insoluble matter at -40 °C until the water content is 0.16 wt% to obtain the lutein.

[0050] Comparative Example 4 A method for extracting lutein from marigold, comprising the following steps: S1 Granulation: The dried marigold flowers are crushed into powder, and then made into fine particles of different specifications of 0.1 - 1.0 mm, 2 - 3 mm, and 3 - 5 mm through a granulator. The 0.1 - 1.0 mm, 2 - 3 mm, and 3 - 5 mm particles are mixed according to a mass ratio of 1:3:2 to obtain marigold flower mixed particles.

[0051] S2 Infiltration: Add 5 times the volume of a mixed solvent to 1 part by mass of the marigold flower mixed particles. The volume ratio of ethanol to ethyl acetate in the mixed solvent is 2:1, and ultrasonic treatment is carried out at 60 °C with 800 W for 1 h to obtain a marigold infiltration solution.

[0052] S3 Extraction: Add 1 part by mass of a mixed solution of betaine and histidine (the mass fraction of the mixed solution is 10%, the mass ratio of chlorogenic acid to histidine is 1:3, and the solvent is water) to 10 parts by mass of the marigold infiltration solution. The pH of the mixed solution is adjusted to 9 with sodium hydroxide, and then stirring extraction is carried out at 40 °C and 250 rpm for 2 h to obtain a marigold extraction solution.

[0053] S4 Concentration: The marigold lutein extraction solution is subjected to vacuum distillation at 60 °C under a pressure of 0.015 MPa until ethanol is no longer detectable. The insoluble matter in the remaining material after distillation is collected to obtain a crude lutein extract. S5 Washing: The crude lutein extract in S4 is washed with 10 times its mass of deionized water, and the washed insoluble matter is freeze-dried at -40 °C until the water content is 0.17 wt% to obtain the lutein.

[0054] Comparative Example 5 A method for extracting lutein from marigold flowers, comprising the following steps: S1 Granulation: The dried marigold flowers are crushed into powder, and then made into fine particles of different specifications of 0.1 - 1.0 mm, 2 - 3 mm, and 3 - 5 mm through a granulator. The 0.1 - 1.0 mm, 2 - 3 mm, and 3 - 5 mm particles are mixed according to a mass ratio of 1:3:2 to obtain marigold flower mixed particles.

[0055] S2 Infiltration: Add 5 times the volume of a mixed solvent to 1 part by mass of the marigold flower mixed particles. The volume ratio of ethanol to ethyl acetate in the mixed solvent is 2:1, and ultrasonic treatment is carried out at 60 °C with 800 W for 1 h to obtain a marigold infiltration solution.

[0056] S3 Extraction: Add 1 part by mass of a mixed solution of chlorogenic acid, betaine, and histidine (the mass fraction of the mixed solution is 10%, the mass ratio of chlorogenic acid, betaine, and histidine is 1:3:3) to 10 parts by mass of the marigold infiltration solution. The pH of the mixed solution is adjusted to 9 with sodium hydroxide, and then stirring extraction is carried out at 40 °C and 250 rpm for 2 h to obtain a marigold extraction solution.

[0057] S4 Concentration: The marigold lutein extract is subjected to vacuum distillation at 60°C under a pressure of 0.015 MPa until ethanol is no longer detectable. The insoluble matter in the remaining distillation material is collected to obtain a crude lutein extract. S5 Washing: The crude lutein extract in S4 is washed with deionized water 10 times its mass. The washed insoluble matter is freeze-dried at -40°C until the water content is 0.17 wt% to obtain the lutein.

[0058] Comparative Example 6 A method for extracting lutein from marigold includes the following steps: S1 Granulation: The dried marigold flowers are crushed into powder, and then made into small particles of different specifications of 0.1 - 1.0 mm, 2 - 3 mm, and 3 - 5 mm through a granulator. The 0.1 - 1.0 mm, 2 - 3 mm, and 3 - 5 mm are in a mass ratio of 1:3:2 to obtain a marigold flower mixed granule.

[0059] S2 Infiltration: Add 5 times the volume of a mixed solvent to 1 part by mass of the marigold flower mixed granule. The volume ratio of ethanol to ethyl acetate in the mixed solvent is 2:1. Ultrasonic at 800 W for 1 h at 60°C to obtain a marigold infiltration solution.

[0060] S3 Extraction: Add 1 part by mass of a mixed solution of chlorogenic acid, betaine, and histidine (the mass fraction of the mixed solution is 10%, and the mass ratio of chlorogenic acid, betaine, and histidine is 1:1:6) to 10 parts by mass of the marigold infiltration solution. The pH of the mixed solution is adjusted to 9 with sodium hydroxide, and then stirred and extracted at 40°C and 250 rpm for 2 h to obtain a marigold extract.

[0061] S4 Concentration: The marigold lutein extract is subjected to vacuum distillation at 60°C under a pressure of 0.015 MPa until ethanol is no longer detectable. The insoluble matter in the remaining distillation material is collected to obtain a crude lutein extract. S5 Washing: The crude lutein extract in S4 is washed with deionized water 10 times its mass. The washed insoluble matter is freeze-dried at -40°C until the water content is 0.17 wt% to obtain the lutein.

[0062] Referring to GB 5009.248 - 2016 National Food Safety Standard - Determination Method of Lutein in Foods, the extraction rates of lutein in the examples and comparative examples are measured respectively, and the purity of lutein is detected by high performance liquid chromatography.

[0063] Extraction rate = [(mass of obtained lutein × purity of lutein) / mass of raw marigold] × 100%. The test results are shown in Table 1.

[0064] Table 1: Test Results

[0065] As can be seen from the table, the extraction rate of lutein by infiltration extraction using marigold granules of a single specification in Comparative Example 1 is lower than that in Example 1 where marigold granules of different specifications are mixed in a certain proportion and then subjected to infiltration extraction of lutein. Comparing Comparative Examples 2-6 with Example 1, it can be seen that by adding chlorogenic acid, betaine, and histidine with a mass ratio of (1-2):1:3 to the infiltration solution for auxiliary extraction, and then collecting and washing the crude lutein product, the extraction rate and purity of lutein in the marigold extraction process can be improved.

[0066] The above embodiments are the preferred embodiments of the present invention, but the embodiments of the present invention are not limited by the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications made without departing from the spirit and principle of the present invention shall be equivalent replacement methods and are all included in the protection scope of the present invention.

Claims

1. A method for extracting lutein from marigold, characterized in that, It includes the following steps: S1 Granulation: The dried marigold flowers are crushed into powder, and then made into fine particles of different specifications of 0.1 - 1.0 mm, 2 - 3 mm, and 3 - 5 mm through a granulator. The particles of different specifications are mixed to obtain marigold mixed particles; S2 Infiltration: One or more solvents in short-chain alkanes or their derivatives are added to the marigold mixed particles for ultrasonic infiltration to obtain marigold infiltration liquid; S3 Extraction: A mixed solution of chlorogenic acid, betaine, and histidine is added to the marigold infiltration liquid for stirring extraction, and then filtered to obtain marigold extract; S4 Concentration: The marigold extract is subjected to reduced-pressure low-temperature distillation until ethanol is undetectable, and the insoluble matter in the remaining distillation material is collected to obtain crude lutein extract; S5 Washing: The crude lutein extract is washed with deionized water, and the washed insoluble matter is freeze-dried at -40 °C to obtain lutein.

2. The extraction method of lutein according to claim 1, characterized in that, For the 0.1 - 1.0 mm, 2 - 3 mm, and 3 - 5 mm particles described in S1 granulation, the mass ratio is 1:(1 - 3):(1 - 2).

3. The extraction method of lutein according to claim 1, characterized in that, For S2, the number of carbon atoms of the short-chain alkane ≤ 6, and the derivative of the alkane is an alcohol, acid, or ester of the alkane. The volume ratio of the solvent to the mass of the marigold mixture particles is 5 - 10:

1.

4. The extraction method of lutein according to claim 1, characterized in that, For S3, the mass ratio of chlorogenic acid, betaine, and histidine is 1 - 2:1:

3.

5. The extraction method of lutein according to claim 1, characterized in that, For S3, the mass fraction of the mixed solution of chlorogenic acid, betaine, and histidine is 10 - 20%, and the pH is 8 - 9.

6. The extraction method of lutein according to claim 1, wherein For S3, the content of the mixed solution of chlorogenic acid, betaine, and histidine in the marigold infiltration liquid is 10 - 20% by mass fraction.

7. The extraction method of lutein according to claim 1, wherein For S4, the conditions for reduced-pressure low-temperature distillation are: 50 °C ≤ temperature ≤ 60 °C.

8. The extraction method of lutein according to claim 1, characterized in that, For S5, the mass ratio of the crude lutein extract to water is 1:10 - 20.

9. Application of marigold lutein prepared by the method according to any one of claims 1 - 8 in animal feed.

10. The use of marigold lutein according to claim 9 in animal feed, characterized in that, The animals described include poultry or aquatic animals.

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