Process for the extraction of caffeic acid and rosmarinic acid from rosemary

Abstract

The present invention relates to a method of extracting caffeic acid and rosmarinic acid from rosemary, and more particularly, to a method of extracting caffeic acid and rosmarinic acid from rosemary including: (S1) a step of preparing an extractor including a main body in which a solution contained in the main body is passed through a filter membrane and the filtered solution is dropped to an outside of a lower portion through a dropping portion connected to the lower portion of the main body, and an aggregator located at a lower portion of the extractor and collecting the filtered solution dropped from the dropping portion; (S2) a step of dipping dried rosemary into the main body and adding a solvent to the main body, and then impregnating the dried rosemary at room temperature; and (S3) a step of dropping the solution after the dried rosemary is impregnated from the dropping portion to the aggregator.

Description

Technical Field

[0001] This invention relates to a method for extracting caffeic acid and rosmarinic acid from rosemary, and more particularly to a method for extracting caffeic acid and rosmarinic acid from rosemary by improving extraction efficiency through an extraction process that is safe for human use without loss of effective components.

[0002] This application claims priority based on Korean Application No. 10-2019-0106894, filed on August 29, 2019, and incorporates all the contents disclosed in the description of the corresponding application. Background Technology

[0003] Rosemary (Rosmarinus officinalis L.), also known as sea dew or rosemary, is a small evergreen shrub belonging to the Lamiaceae family. Its leaves, buds, and flowers are edible, and it is a temperate perennial native to Southern Europe. As a perennial with needle-like leaves, it is an herbaceous plant native to the Mediterranean coast, possessing green leaves and a distinctive fragrance. The flowers bloom in winter and spring, and come in white, pink, purple, and blue. It retains its aroma even when heated, so it is often used in cooking meat dishes with sage or thyme. Due to its strong aroma, it is recommended to use it sparingly to preserve the flavor of the main dish. As a major component, it contains a wide variety of effective ingredients such as α-pinene, apigenin, β-carotene, β-sitosterol, betulinic acid, borneol, caffeic acid, capersana, carawayol, carvone, caryophyllene, chlorogenic acid, diosmin, genistein, geraniol, hesperidin, limonene, linalool, 1,8-cineole, rosmarinic acid, salicylic acid, squalene, tannins, thymol, ursolic acid, calcium, magnesium, manganese, phosphorus, potassium, zinc, vitamin B1, vitamin B3, and vitamin C. It is currently known to possess antibacterial activity (Registered Patent No. 10-0729182, No. 1...). It has multiple application possibilities, including: 0-0362885, 10-1161415, antioxidant activity (registered patents 10-1205680, 10-1205867, 10-1454696, 10-0606649, 10-0971655), whitening activity (registered patent 10-1513237), atopic dermatitis reliever (registered patent 10-1236946), and antiviral properties (registered patent 10-0697309, application patent 10-2015-0045259).

[0004] In particular, numerous studies have demonstrated that caffeic acid has effects such as anti-inflammation, reducing skin cancer, and inhibiting the growth of oral cancer cells. Furthermore, rosmarinic acid has been shown to have powerful antioxidant, anti-allergic, immunosuppressive, and atopic-related effects. For example, Korean Patent No. 10-1576232 discloses a method for extracting caffeic acid and rosmarinic acid from seaweed by adding a eutectic solvent.

[0005] However, there is still a need to develop an extraction method that is safe for human use and can effectively separate caffeic acid and rosmarinic acid. Summary of the Invention

[0006] Therefore, the purpose of this invention is to provide a method for extracting caffeic acid and rosmarinic acid from rosemary by improving extraction efficiency through an extraction process that is safe for the human body without loss of effective components.

[0007] To achieve the objectives described above, the method for extracting caffeic acid and rosmarinic acid from rosemary according to the present invention comprises: (S1) preparing an extraction apparatus comprising an extractor that allows a solution contained in a main body to pass through a filter membrane and drip through a dripping portion connected to the lower part of the main body to the lower outer side of the main body, and an aggregator located at the lower part of the extractor and collecting the filtered solution dripping from the dripping portion; (S2) adding dried rosemary to the main body and adding a solvent to the main body, and then impregnating the dried rosemary at room temperature; and (S3) dripping the solution after impregnating the dried rosemary from the dripping portion into the aggregator.

[0008] At this time, the extraction device in step (S1) may further include: a water storage tank, which replenishes the main body with new solution according to the decrease in the amount of solution contained in the main body.

[0009] Furthermore, the solvent may comprise one or a mixture of two or more of the following selected from the group consisting of monohydric alcohols of C1 to C5, dihydric alcohols of C2 to C5, and trihydric alcohols of C3 to C5.

[0010] At this point, the solvent may also contain water.

[0011] Furthermore, after step (S3), the step of transferring the solution contained in the aggregator to the main body may also be included.

[0012] In this invention, by employing an extraction method in which the extraction solution is dripped from the top to the bottom, the loss of active ingredients due to heat can be prevented when extracting caffeic acid and rosmarinic acid from rosemary, and the extraction efficiency can be improved through an extraction process that is safe for the human body. Attached Figure Description

[0013] Figure 1 This is a schematic diagram illustrating an engineering process for extracting caffeic acid and rosmarinic acid from rosemary according to one embodiment of the present invention.

[0014] Figure 2 This is a schematic diagram illustrating a schematic illustration of an apparatus for extracting caffeic acid from rosemary and an extraction apparatus for rosmarinic acid according to one embodiment of the present invention.

[0015] Figure 3 This is a schematic diagram illustrating the extraction of caffeic acid from rosemary and the extraction apparatus for rosmarinic acid according to another embodiment of the present invention.

[0016] [Symbol Explanation]

[0017] 10: Extractor

[0018] 11: Main body

[0019] 12: Filter membrane

[0020] 13: Dripping section

[0021] 20: Aggregator

[0022] 30: Reservoir

[0023] 100: Extraction device Detailed Implementation

[0024] The embodiments of the present invention will now be described in detail with reference to the accompanying drawings, so that those skilled in the art can easily implement the invention. However, the present invention can be implemented in many different forms and is not limited to the embodiments and drawings described herein.

[0025] This invention was developed during research on low-temperature extraction methods to more safely and effectively extract the physiologically active components of heat-sensitive natural substances. It focused on the fact that even when using the same coffee beans, a cold extraction process with long-term cold water extraction can increase the caffeine content and reduce bitterness.

[0026] Next, a method for extracting caffeic acid and rosmarinic acid from rosemary according to one embodiment of the present invention will be described in detail with reference to the accompanying drawings.

[0027] Figure 1 This is a schematic diagram illustrating an engineering process for extracting caffeic acid and rosmarinic acid from rosemary according to one embodiment of the present invention. Figure 2 as well as Figure 3 This is a schematic diagram illustrating a schematic illustration of an apparatus for extracting caffeic acid from rosemary and an extraction apparatus for rosmarinic acid according to one embodiment of the present invention.

[0028] See Figures 1 to 3 First, prepare an extraction apparatus for caffeic acid and rosmarinic acid from rosemary (step S1).

[0029] The extraction device 100 is as follows Figure 2 As shown, an extractor 10 is provided at the upper part, and an aggregator 20 is provided at the lower part of the extractor 10.

[0030] The extractor 10 allows the solution contained in the main body 11 to pass through the filter membrane 12 and drip through the dripping part 13 connected to the lower part of the main body 11 to the lower outer side.

[0031] The extractor 10 is a reactor that extracts active ingredients from solid substances, such as dried rosemary or rosemary powder, by storing them with a solvent. It includes a filter membrane 12 to prevent loss of the solid substance and to filter the extracted solution. Preferably, a stopcock valve may be provided in the dripping section 13 to regulate the dripping rate of the filtered solution containing the active ingredients.

[0032] The aggregator 20, located below the extractor 10, is used to aggregate a filtered solution containing the active ingredient of rosemary that drips from the dripping section 13, and its type is not limited in any way.

[0033] Furthermore, in order to improve the extraction efficiency of the original material located inside the main body 11, it is particularly important to maintain the extraction solvent at a certain level. Therefore, it is advisable to continuously replenish the main body 11 with new solvent based on the amount reduced when the solvent contained in the main body 11 containing the active ingredient is discharged downwards. For this purpose, a water storage tank 30 for storing the new solvent may also be included, such as... Figure 3 As shown, the water reservoir 30 can be located above the extractor 10. To control the drip rate of the new solvent, the water reservoir 30 preferably includes a stopcock valve.

[0034] Next, the dried rosemary is added to the main body 11 and a solvent is added to the main body 11, and then the dried rosemary is impregnated at room temperature (step S2).

[0035] To improve extraction efficiency, the dried rosemary can be cut into a certain size or powdered.

[0036] When extracting the dried rosemary, the extraction efficiency of caffeic acid and rosmarinic acid can be improved by immersing the rosemary and solvent in the main body 11 at room temperature.

[0037] Impregnation is a process of immersing the solvent into the cell membrane of the dried material to induce swelling. Compared to extraction performed directly in the dried state, extraction after impregnation can significantly improve efficiency.

[0038] In addition, the solvent may be an alcohol, specifically one or a mixture of two or more of the following selected from the group consisting of monohydric alcohols of C1 to C5, dihydric alcohols of C2 to C5, and trihydric alcohols of C3 to C5.

[0039] The monohydric alcohols may be methanol, ethanol, propanol, isopropanol, butanol, tert-butanol, and pentanol; the dihydric alcohols may be ethylene glycol, propylene glycol, butanediol, and pentanediol; and the trihydric alcohols may be glycerol, but their types are not specifically limited.

[0040] When using diols and triols other than ethanol alone, no caffeic acid or rosmarinic acid was detected in conventional room temperature maceration methods that involve extraction over 24 hours. However, when extraction was performed according to the present invention, significantly higher amounts of caffeic acid and rosmarinic acid were detected.

[0041] Furthermore, in existing room temperature impregnation methods, when the solvent is water or the water content in the solvent is more than half, such as when the water content is 80% by weight or more, no caffeic acid and rosmarinic acid are detected even when stirring. However, in the method according to the present invention, when using a solvent that mixes water and the alcohol, even when the water content in the solvent is more than half, such as when it is 80% by weight or more, a large amount of caffeic acid and rosmarinic acid are detected.

[0042] Next, the solution used to impregnate the dried rosemary is dripped from the drip section 13 into the collector 20 (step S3).

[0043] When the dried rosemary contains sufficient extraction solvent, the solution can be dripped into the collector through the drip section 13 located at the bottom of the extractor 10. At this time, drip extraction can be performed while adjusting the dripping rate by opening the piston valve located in the drip section 13, and the dripping solvent will be collected in the collector.

[0044] At this point, after step (S3), the solution contained in the aggregator 20 can be transferred to the main body 11, and then drip extraction can be performed again. The solution contained in the aggregator 20 can be transferred to the main body 11, and further transferred to the water storage tank 30. Through the process described above, the extraction efficiency of caffeic acid and rosmarinic acid can be further improved.

[0045] As described above, the method of the present invention for extracting caffeic acid and rosmarinic acid from rosemary can minimize the loss of heat-sensitive active ingredients by extracting at room temperature, and is a very effective method for extracting caffeic acid and rosmarinic acid using alcohols or mixed solvents of alcohols and water.

[0046] The present invention will now be described in more detail with reference to specific embodiments. The following embodiments are merely illustrative and are not intended to limit the invention.

[0047] Example: Droplet Extraction

[0048] As an extractor that allows the solution contained in the main body to pass through a filter membrane and drip through a dripping section connected to the lower part of the main body to the outside of the lower part, an extractor with adjustable dripping rate of solvent is installed at the upper part, and an aggregator is installed at the lower part to collect the filtered solution dripping from the dripping section of the extractor.

[0049] 50g of dried rosemary and 950g of solvent as described in Table 2 were added to the main body of the extractor prepared in the manner described. After soaking at room temperature for 1 hour to allow the dried rosemary to fully absorb the solvent, the extractor solution was slowly dripped into the lower collector for 30 minutes to obtain the extract.

[0050] For reference, Example 6a in Table 2 below is a case where the extract obtained in Example 5c is put back into the extractor and extraction is performed again (extraction twice), while Example 6b is a case where the extract obtained in Example 6a is put back into the extractor and extraction is performed again (extraction three times).

[0051] Comparative example: maceration extraction

[0052] 50g of dried rosemary and 950g of the solvents listed in Table 2 below were placed in a general container and soaked at room temperature for 24 hours. The rosemary extract was obtained by filtration.

[0053] Experimental Example: Compositional Analysis of Extract

[0054] The content of the components was confirmed under the following analytical conditions.

[0055] The standard solution was prepared by dissolving caffeic acid and rosmarinic acid standards in the mobile phase of high performance liquid chromatography (HPLC) to a concentration of 100 ppm, and then filtering it through a 0.45 μm thick polyvinylidene fluoride (PVDF) membrane filter.

[0056] Sample solutions for the examples and comparative examples were prepared by filtering the rosemary extract stock solution (maceration extract and dropwise extract) through a 0.45 μm thick filter and analyzed according to the high performance liquid chromatography (HPLC) analysis conditions described below. The results are shown in Table 2.

[0057] 1. Analytical apparatus

[0058] High Performance Liquid Chromatography (Waters 2695, 2998 PDA detector)

[0059] 2. Analysis conditions

[0060] (1) Column: C18, 4.6*250mm, 5μm

[0061] (2) Flow rate: 1 ml / min

[0062] (3) Injection volume: 10 μl

[0063] (4) Detection: 327nm (rosmarinic acid), 323nm (caffeic acid)

[0064] (5) Mobile phase

[0065] Table 1

[0066]

[0067] (6) Standard: Rosmarinic acid, caffeic acid

[0068] The components of the embodiments and comparative examples described in Table 2 below were analyzed according to the analytical conditions described above, and the results are shown in Table 2 below.

[0069] Table 2

[0070]

[0071] As shown in Table 2 above, a comparison of Comparative Examples 2 to 4 using the conventional maceration extraction method and Examples 1, 4, and 5c using the drip extraction method of the present invention reveals that, using the same solvent, the drip extraction method of the present invention yields higher levels of caffeic acid and rosmarinic acid. While no caffeic acid or rosmarinic acid was extracted using the conventional maceration extraction method (Comparative Example 4), a considerable amount of caffeic acid and rosmarinic acid were extracted in Example 5c using the same solvent. This confirms that the drip extraction method is significantly more effective than the conventional maceration extraction method.

[0072] Furthermore, in Comparative Example 1, where rosemary was impregnated in water as a solvent, and in Comparative Example 4, where a solvent containing 80% by weight of water was used for impregnation extraction, no caffeic acid or rosmarinic acid was extracted. However, in Examples 5b and 5c, even with a higher proportion of water, a considerable amount of caffeic acid and rosmarinic acid could be extracted, thereby confirming that the drip extraction method of the present invention is a very effective method.

[0073] Furthermore, observations of Examples 5a to 5c, which compared the amounts of water and alcohol used, revealed that even with an increase in water content, the caffeic acid content did not decrease; on the contrary, it tended to increase.

[0074] Furthermore, observations of Examples 6a and 6b, in which the extraction steps were repeated on the extract obtained in Example 5c, confirmed that the content of caffeic acid and rosmarinic acid increased significantly with the increase in the number of extractions.

[0075] Therefore, it can be confirmed that the drip extraction method of the present invention can very effectively extract caffeic acid and rosmarinic acid from rosemary.

[0076] The foregoing description is merely an illustrative description of the present invention. Those skilled in the art should understand that the present invention can be implemented in different modifications without departing from its essential characteristics. Therefore, the disclosed embodiments and experimental examples should be understood as illustrative rather than limiting. The scope of the present invention should be defined by the claims rather than the foregoing description, and all differences from its equivalents should be interpreted as being included within the scope of the present invention.

Claims

1. A method of extracting caffeic acid and rosmarinic acid from rosemary, characterized by, Comprising: (S1) a step of preparing an extractor including passing a solution contained in a main body through a filtration membrane and dropping the filtered solution to an outside of a lower portion through a dropping portion connected to a lower portion of the main body, and an accumulator positioned at a lower portion of the extractor and collecting the filtered solution dropped from the dropping portion; (S2) a step of putting dried rosemary into the main body, adding a solvent to the main body, and then impregnating the dried rosemary at normal temperature for 1 hour; and, (S3) a step of dropping the solution after impregnating the dried rosemary from the dropping portion to the accumulator, wherein the solvent is one or a mixture of two or more selected from the group consisting of C1 to C5 monohydric alcohols, C2 to C5 dihydric alcohols, and C3 to C5 trihydric alcohols.

2. The method of extracting caffeic acid and rosmarinic acid from rosemary according to claim 1, characterized in that: the extractor in the (S1) step further includes a water reservoir that replenishes the main body with a new solution according to a decrease in the solution contained in the main body.

3. The method of extracting caffeic acid and rosmarinic acid from rosemary according to claim 1, characterized in that: the solvent further contains water.

4. The method of extracting caffeic acid and rosmarinic acid from rosemary according to claim 1, characterized in that: after the (S3) step, a step of transferring the solution contained in the accumulator to the main body is further included.

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