Method for extracting chlorogenic acid from desert cistanche

Abstract

The invention discloses a method for extracting chlorogenic acid from cistanche deserticola, which comprises the following steps: freeze-drying the cistanche deserticola, crushing into powder, adding an ethanol solution, and carrying out reflux extraction to obtain a cistanche deserticola mixed solution; centrifuging the cistanche deserticola mixed solution, and then filtering and separating to obtain filtrate I and filter residue I; soaking the filter residue I in an ethanol solution, adding a compound enzyme for enzymolysis, and performing centrifugal filtration to obtain a filtrate II and a filter residue II; combining the filtrate I and the filtrate II to obtain a chlorogenic acid extracting solution, and purifying the chlorogenic acid extracting solution through macroporous resin to obtain the desert cistanche chlorogenic acid. According to the method, the chlorogenic acid component in the desert cistanche can be effectively extracted, and desert cistanche biological resources are fully utilized.

Description

Technical Field

[0001] The invention relates to the technical field of food processing, and in particular to a method for extracting chlorogenic acid from Cistanche deserticola. Background Art

[0002] Chlorogenic acid is an important class of phenolic acid secondary metabolites, which is widely present in the plant kingdom. Chlorogenic acid plays an important role in plant growth and development, and in resisting biotic and abiotic stresses. In addition, it has a variety of biological activities and pharmacological functions, and has important application potential in anti-inflammatory, antibacterial and hypoglycemic aspects, and is a potential functional food ingredient. However, the content of chlorogenic acid in plants is usually very low, which seriously restricts its development and utilization value.

[0003] It is a perennial herb of the genus Cistanche of the family Orobanchaceae. It is also known as Dayun, Sweet Dayun, Cistanche, Sweet Cistanche, Goblin, etc. Its dried scaly leaves and fleshy stems are precious Chinese medicinal materials. It has a long history of use since ancient times and is known as the "Desert Ginseng". Modern pharmacological studies have shown that Desert Cistanche has the effects of moistening the intestines and promoting bowel movements, protecting the liver, regulating immunity, resisting osteoporosis, resisting tumors, improving memory, resisting oxidation, resisting aging, resisting fatigue, promoting wound healing, protecting ischemic myocardium, and improving microcirculation. However, due to long-term over-exploitation, the wild resources of Desert Cistanche have been sharply reduced, and it has been listed as a second-level protected plant in my country. Moreover, there are a lot of confusing varieties of Cistanche deserticola on the market, including Cistanche tubulosa, Cistanche sanderiana, Cistanche halogenide, Cistanche lanzhouensis, and the fake Cistanche multistachys. Chlorogenic acid and its content vary greatly from different origins and parts; different extraction process seriously affect the composition and proportion of the chlorogenic acid extract of Cistanche deserticola.

[0004] Therefore, a method for extracting chlorogenic acid from Cistanche deserticola is needed to effectively extract chlorogenic acid from Cistanche deserticola, thereby making full use of the biological resources of Cistanche deserticola. Summary of the invention

[0005] In order to solve the above problems, the present invention provides a method for extracting chlorogenic acid from Desert Cistanche deserticola, which can effectively extract the chlorogenic acid component in Desert Cistanche deserticola and make full use of the biological resources of Desert Cistanche deserticola.

[0006] To achieve the above object, the present invention proposes a method for extracting chlorogenic acid from Cistanche deserticola, which comprises the following steps:

[0007] (1) freeze-drying and crushing the deserticola Cistanche into powder, adding ethanol solution, and refluxing extraction to obtain a deserticola Cistanche mixed solution;

[0008] (2) centrifuging the deserticola mixed solution and filtering to obtain filtrate I and filter residue I;

[0009] (3) soaking the filter residue I in an ethanol solution, adding a composite enzyme for enzymatic hydrolysis, and centrifugally filtering to obtain a filtrate II and a filter residue II;

[0010] (4) The filtrate I and the filtrate II are combined to obtain a chlorogenic acid extract, and the chlorogenic acid extract is purified by a macroporous resin to obtain desert Cistanche deserticola chlorogenic acid.

[0011] According to an embodiment of the present invention, a method for extracting chlorogenic acid from Cistanche deserticola is provided. The method utilizes reflux extraction and complex enzyme enzymatic hydrolysis to more effectively break the cell wall of Cistanche deserticola, so that the effective ingredients can be better dissolved, thereby improving the extraction rate of chlorogenic acid from Cistanche deserticola; through the use of macroporous resin, impurities in the extract can be effectively removed, the purity of chlorogenic acid from Cistanche deserticola is improved, and Cistanche deserticola is more fully utilized, which is conducive to promoting the development and application of Cistanche deserticola resources.

[0012] Optionally, in step (1), the freeze-drying temperature is -20°C to -60°C, and the freeze-drying time is 4 to 12 hours.

[0013] Optionally, in step (1), the volume fraction of the ethanol solution is 60% to 90%; and the solid-liquid ratio of the Cistanche deserticola powder to the ethanol solution is 1:10 to 1:25 g / mL.

[0014] Optionally, in step (1), the temperature of the reflux extraction is 60 to 90° C., and the time of the reflux extraction is 0.5 to 2.5 h.

[0015] Optionally, in step (2), the centrifugal speed is 6000-12000 rpm / min, the centrifugal time is 1-10 min; and the filtration is ceramic membrane filtration.

[0016] Optionally, in step (3), the volume fraction of the ethanol solution is 60% to 90%, and the solid-liquid ratio of the filter residue I to the ethanol solution is 1:10 to 1:25 g / mL.

[0017] Optionally, in step (3), the complex enzyme is pectin lyase and hemicellulase; the enzymatic hydrolysis temperature is 50-60° C., and the enzymatic hydrolysis time is 0.5-1.5 h.

[0018] Furthermore, the added amount of the pectin lyase is 0.3-0.6 wt % of the mixed solution; and the added amount of the hemicellulase is 0.5-1.5 wt % of the mixed solution.

[0019] Optionally, in step (4), the macroporous resin is macroporous resin ADS-7.

[0020] Furthermore, in step (5), the purification process includes water washing to remove impurities and elution with ethanol solution; the volume of the water washing is 2 to 4 BV; the flow rate of the water washing is 2 to 3 BV / h; the volume fraction of the ethanol solution is 60 to 80%; the volume of the ethanol solution elution is 3 to 6 BV; the flow rate of the ethanol solution elution is 2 to 4 BV / h.

[0021] Additional aspects and advantages of the present invention will be given in part in the following description and in part will be obvious from the following description, or will be learned through practice of the present invention. DETAILED DESCRIPTION

[0022] The technical solution of the present invention is described below through specific examples. It should be understood that the one or more method steps mentioned in the present invention do not exclude the existence of other method steps before and after the combination step or the insertion of other method steps between these explicitly mentioned steps; it should also be understood that these embodiments are only used to illustrate the present invention and are not used to limit the scope of the present invention. Moreover, unless otherwise specified, the numbering of each method step is only a convenient tool for identifying each method step, and is not intended to limit the order of arrangement of each method step or to limit the scope of the present invention. The change or adjustment of the relative relationship thereof shall also be regarded as the scope of the present invention without substantially changing the technical content.

[0023] In order to better understand the above technical scheme, the exemplary embodiments of the present invention are described in more detail below. Although exemplary embodiments of the present invention are shown, it should be understood that the present invention can be implemented in various forms and should not be limited by the embodiments set forth herein. On the contrary, these embodiments are provided in order to enable a more thorough understanding of the present invention and to enable the scope of the present invention to be fully communicated to those skilled in the art.

[0024] The test materials used in the present invention are all common commercial products and can be purchased in the market.

[0025] The present invention is described below with reference to specific embodiments. It should be noted that these embodiments are merely illustrative and do not limit the present invention in any way.

[0026] Example 1

[0027] A method for extracting chlorogenic acid from Cistanche deserticola, comprising the following steps:

[0028] (1) After freeze-drying the deserticola Cistanche at -40°C for 12 hours, the powder was crushed into powder, passed through a 60-mesh sieve, and mixed with 80% ethanol solution at a solid-liquid ratio of 1:15 g / mL, and refluxed at 80°C for 1.5 hours to obtain a deserticola Cistanche mixed solution;

[0029] (2) The mixed solution of Cistanche deserticola was centrifuged at 7000 rpm / min for 5 min and then filtered through a ceramic membrane to obtain filtrate I and residue I;

[0030] (3) Soaking the residue I in an 80% ethanol solution at a solid-liquid ratio of 1:15 g / mL to obtain a mixed solution, adding 0.5 wt% pectin lyase and 1.0 wt% hemicellulase to the mixed solution, performing enzymolysis at 50°C for 1.0 h, then inactivating the enzyme at 90°C for 15 min, centrifuging at 7000 rpm / min for 5 min, and separating by filtering with a ceramic membrane to obtain a filtrate II and a residue II;

[0031] (4) Filtrate I and filtrate II were combined to obtain a chlorogenic acid extract, which was purified by macroporous resin ADS-7. The extract was washed with 4 BV of ultrapure water at a flow rate of 3 BV / h, and eluted with 6 BV of 60% ethanol solution at a rate of 2 BV / h. The eluate was collected, concentrated and dried to obtain the chlorogenic acid of Cistanche deserticola. The extraction rate was 4.21%, and the purity of the chlorogenic acid of Cistanche deserticola was 97.31%.

[0032] Example 2

[0033] A method for extracting chlorogenic acid from Cistanche deserticola, comprising the following steps:

[0034] (1) After freeze-drying the deserticola Cistanche at -40°C for 12 hours, the powder was crushed into powder, passed through a 60-mesh sieve, and mixed with a 60% ethanol solution at a solid-liquid ratio of 1:25 g / mL, and refluxed at 80°C for 1.5 hours to obtain a deserticola Cistanche mixed solution;

[0035] (2) The mixed solution of Cistanche deserticola was centrifuged at 7000 rpm / min for 5 min and then filtered through a ceramic membrane to obtain filtrate I and residue I;

[0036] (3) Soaking the residue I in a 60% ethanol solution at a solid-liquid ratio of 1:25 g / mL to obtain a mixed solution, adding 0.5 wt% pectin lyase and 1.0 wt% hemicellulase to the mixed solution, performing enzymolysis at 50°C for 1.0 h, then inactivating the enzyme at 90°C for 15 min, centrifuging at 7000 rpm / min for 5 min, and separating by filtering with a ceramic membrane to obtain a filtrate II and a residue II;

[0037] (4) Filtrate I and filtrate II were combined to obtain a chlorogenic acid extract, which was purified by macroporous resin ADS-7. The extract was washed with 4 BV of ultrapure water at a flow rate of 3 BV / h, and eluted with 6 BV of 60% ethanol solution at a rate of 2 BV / h. The eluate was collected, concentrated and dried to obtain the chlorogenic acid of Cistanche deserticola. The extraction rate was 3.57%, and the purity of the chlorogenic acid of Cistanche deserticola was 97.64%.

[0038] Example 3

[0039] A method for extracting chlorogenic acid from Cistanche deserticola, comprising the following steps:

[0040] (1) After freeze-drying the deserticola Cistanche at -40°C for 12 hours, the powder was crushed into powder, passed through a 60-mesh sieve, and mixed with 80% ethanol solution at a solid-liquid ratio of 1:15 g / mL, and refluxed at 80°C for 1.5 hours to obtain a deserticola Cistanche mixed solution;

[0041] (2) The mixed solution of Cistanche deserticola was centrifuged at 7000 rpm / min for 5 min and then filtered through a ceramic membrane to obtain filtrate I and residue I;

[0042] (3) Soaking the residue I in an 80% ethanol solution at a solid-liquid ratio of 1:15 g / mL to obtain a mixed solution, adding 0.5 wt% pectin lyase and 0.5 wt% hemicellulase to the mixed solution, performing enzymolysis at 50°C for 0.5 h, then inactivating the enzyme at 90°C for 15 min, centrifuging at 7000 rpm / min for 5 min, and separating by filtering with a ceramic membrane to obtain filtrate II and residue II;

[0043] (4) Filtrate I and filtrate II were combined to obtain a chlorogenic acid extract, which was purified by macroporous resin ADS-7. The extract was washed with 4 BV of ultrapure water at a flow rate of 3 BV / h, and eluted with 6 BV of 60% ethanol solution at a rate of 2 BV / h. The eluate was collected, concentrated and dried to obtain the chlorogenic acid of Cistanche deserticola. The extraction rate was 3.42%, and the purity of the chlorogenic acid of Cistanche deserticola was 96.54%.

[0044] Comparative Example 1

[0045] The difference between Comparative Example 1 and Example 1 is that in step (3), only 1.5 wt% of pectin lyase is added to the mixed solution for enzymatic hydrolysis. The extraction rate of chlorogenic acid from Cistanche deserticola was 2.16% and the purity was 95.87%.

[0046] Comparative Example 2

[0047] The difference between Comparative Example 2 and Example 1 is that in step (3), only 1.5 wt% of the mixed solution is enzymatically hydrolyzed by hemicellulase. The extraction rate of chlorogenic acid from Cistanche deserticola was 2.63% and the purity was 96.30%.

[0048] Comparative Example 3

[0049] The difference between Comparative Example 3 and Example 1 is that in step (4), the macroporous resin ADS-7 is replaced by macroporous resin D101. The extraction rate of chlorogenic acid from Cistanche deserticola is 3.17% and the purity is 89.58%.

[0050] In summary, according to the embodiments of the present invention, the present invention has established a method for extracting chlorogenic acid from Cistanche deserticola, so that Cistanche deserticola can be more fully utilized, which is conducive to promoting the efficient development and application of Cistanche deserticola resources. Compared with simple reflux extraction, the addition of composite enzyme hydrolysis can more effectively break the cell wall of Cistanche deserticola, so that the effective ingredients are better dissolved, the extraction rate of chlorogenic acid from Cistanche deserticola is improved, and the purity of the chlorogenic acid finally obtained is also higher, thereby making full use of the biological resources of Cistanche deserticola.

[0051] In the description of this specification, the description with reference to the terms "one embodiment", "some embodiments", "example", "specific example", or "some examples" etc. means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representation of the above terms should not be understood as necessarily being directed to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described may be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art may combine and combine different embodiments or examples described in this specification.

[0052] Although the embodiments of the present invention have been shown and described above, it is to be understood that the above embodiments are exemplary and are not to be construed as limitations of the present invention. A person skilled in the art may change, modify, replace and vary the above embodiments within the scope of the present invention.

Claims

1. A method for extracting chlorogenic acid from Cistanche deserticola, characterized in that: The following steps are involved: (1) freeze-drying and crushing the deserticola Cistanche into powder, adding ethanol solution, and refluxing extraction to obtain a deserticola Cistanche mixed solution; (2) centrifuging the deserticola mixed solution and filtering to obtain filtrate I and filter residue I; (3) soaking the filter residue I in an ethanol solution, adding a composite enzyme for enzymatic hydrolysis, and centrifugally filtering to obtain a filtrate II and a filter residue II; (4) The filtrate I and the filtrate II are combined to obtain a chlorogenic acid extract, and the chlorogenic acid extract is purified by a macroporous resin to obtain desert Cistanche deserticola chlorogenic acid.

2. The method according to claim 1, characterized in that In step (1), the freeze-drying temperature is -20°C to -60°C, and the freeze-drying time is 4 to 12 hours.

3. The method according to claim 1, characterized in that In step (1), the volume fraction of the ethanol solution is 60% to 90%; the solid-liquid ratio of the deserticola powder to the ethanol solution is 1:10 to 1:25 g / mL.

4. The method according to claim 1, characterized in that In step (1), the temperature of the reflux extraction is 60 to 90° C., and the time of the reflux extraction is 0.5 to 2.5 h.

5. The method according to claim 1, characterized in that In step (2), the centrifugal speed is 6000-12000 rpm / min, and the centrifugal time is 1-10 min; and the filtration is ceramic membrane filtration.

6. The method according to claim 1, characterized in that In step (3), the volume fraction of the ethanol solution is 60% to 90%, and the solid-liquid ratio of the filter residue I to the ethanol solution is 1:10 to 1:25 g / mL.

7. The method according to claim 1, characterized in that In step (3), the complex enzyme is pectin lyase and hemicellulase; the enzymatic hydrolysis temperature is 50-60° C., and the enzymatic hydrolysis time is 0.5-1.5 h.

8. The method according to claim 7, characterized in that The added amount of the pectin lyase is 0.3-0.6 wt % of the mixed solution; the added amount of the hemicellulase is 0.5-1.5 wt % of the mixed solution.

9. The method according to claim 1, characterized in that In step (4), the macroporous resin is macroporous resin ADS-7.

10. The method according to claim 9, characterized in that In step (5), the purification process includes water washing to remove impurities and elution with ethanol solution; the volume of the water washing is 2 to 4 BV; the flow rate of the water washing is 2 to 3 BV / h; the volume fraction of the ethanol solution is 60 to 80%; the volume of the ethanol solution elution is 3 to 6 BV; the flow rate of the ethanol solution elution is 2 to 4 BV / h.