KR101903630B1 - Method for extracting essential oils and polyphenols of pines consectively - Google Patents

Abstract

The present invention relates to a processing method of pine trees for continuously extracting essential oils and polyphenol fractions from pine trees.

Description

FIELD OF THE INVENTION [0001] The present invention relates to a method for continuously extracting essential oils and polyphenol fractions from pine trees,

The present invention relates to a processing method of pine trees for continuously extracting essential oils and polyphenol fractions from pine trees.

Essential oils obtained from pine trees are used in a variety of fields such as fragrances, bath salts such as fragrances and soaps. However, in the extraction of essential oil and polyphenol component of pine tree, since the process is separately performed, the polyphenol fraction is inevitably discarded when only essential oil is extracted, and there is a problem that essential oil can not be used to obtain polyphenol fraction .

Accordingly, the present inventors have developed a process capable of continuously obtaining polyphenols and polysaccharides from the remaining residues by extracting essential oil using pine leaves, twigs, and bark removed during the cutting process, , And a polysaccharide can be obtained, thereby completing the present invention.

Korea Patent Publication No. 2007-0111627

The object of the present invention is to continuously obtain refinery oil and polyphenol from pine trees.

In order to achieve the above object,

(a) steam distilling the pine tree to produce an essential oil; And

(b) removing the essential oil from the obtained distillate and subjecting the remaining material to hot water extraction to obtain polyphenol, wherein the essential oil and the polyphenol are continuously extracted from the pine tree .

The present invention also relates to a polyphenol obtained by the process,

0.7% by weight or more of total flavonoids;

10% by weight or more of total proanthocyanidin; And

Wherein the polyphenol comprises at least 30% by weight of the total polyphenol.

The processing method of the present invention can extract polyphenols and polysaccharides continuously by extracting pine essential oil and then processing the residue that is discarded, thereby saving time, labor and energy, and saving resources .

BRIEF DESCRIPTION OF THE DRAWINGS Fig. 1 is a flowchart showing a processing method according to the present invention. The left side is a process using steam distillation and the right side is a process using hydro distillation.
Figure 2 is the yield of the polyphenol fraction obtained using steam distillation or hydro distillation, respectively.
FIG. 3 shows results of ABTS antioxidation experiments on the polyphenol fractions obtained by using steam distillation or hydro distillation, respectively.
FIG. 4 shows results of DPPH antioxidation experiments on the polyphenol fractions obtained using steam distillation or hydro distillation, respectively.
FIG. 5 shows the results of an experiment in which the total flavonoid content was measured for polyphenol fractions obtained by using steam distillation or hydro distillation, respectively.
FIG. 6 shows the results of an experiment in which the total amount of proanthocyanidin was measured for polyphenol fractions obtained by using steam distillation or hydro distillation, respectively.
FIG. 7 shows the results of an experiment in which the total polyphenol content was measured for polyphenol fractions obtained by using steam distillation or hydro distillation, respectively.

The present invention, in one aspect,

(a) steam distilling the pine tree to produce an essential oil; And

(b) removing the essential oil from the obtained distillate and subjecting the remaining material to hot water extraction to obtain polyphenol. The present invention also relates to a method for continuously extracting essential oils and polyphenols from pine trees.

In the present specification, the steam distillation method is a distillation method in which hot steam is directly applied to a sample, and the essential oil evaporated by the hot steam is condensed by using a condenser. In order to better separate the distilled essential oil and water, a fractionating funnel containing a small amount of n-hexane may be placed on the falling portion to collect the essential oil in the nucleic acid layer. In order to remove moisture contained in the essential oil hexane layer, And the like.

The present invention, in another aspect,

(a) hydro-distilling the pine tree to produce an essential oil; And

(b) removing the essential oil from the obtained distillate and subjecting the remaining material to hot water extraction to obtain polyphenol. The present invention also relates to a method for continuously extracting essential oils and polyphenols from pine trees.

In the present specification, the hydro-distillation method is a method in which a sample is placed in a container filled with distilled water and boiled, and an essential oil component distilled due to high temperature is collected using a condenser to produce essential oil. This method is a good way to extract the essential oils that are weak at high temperatures because the water does not rise above the boiling point of the water because the water touches the sample directly.

The continuous extraction method according to one aspect of the present invention not only shows that polyphenol can be continuously obtained by using residue after extraction of essential oil which has been discarded but also polyphenol obtained from the residue after extracting essential oil has excellent antioxidant ability .

In the method of continuously extracting essential oil and polyphenol from pine, which is one aspect of the present invention, the pine tree may be a sample having a size of 8 mesh or less. When the pine sample has a size of 8 mesh or less, it was confirmed that the extraction ratio of essential oil was excellent. In view of the above, in the method of continuously extracting essential oil and polyphenol from pine, which is one aspect of the present invention, the pine has a diameter of 7.5 mesh or less, 7 mesh or less, 6.5 mesh or less , 6 mesh or less, or 5.5 mesh or less. Specifically, the pine sample has a size of 0.1 mesh or more. A mesh is an eye of the mesh, usually represented by the number of meshes contained in a square of 1 inch (25.4 mm) on one side.

In the method of continuously extracting essential oil and polyphenol from pine, which is one aspect of the present invention, the step (a)

(a) separating the essential oil with hexane. Specifically, the essential oil obtained by the hydro-distillation method can be separated from the essential oil by using a hexane / water mixture.

In one aspect of the present invention, a method for continuously extracting essential oils and polyphenols from pine trees, the pine trees may comprise one or more samples selected from the group consisting of barks, leaves and branches of pine trees.

In a method for continuously extracting essential oil and polyphenol from pine, which is one aspect of the present invention, the pine tree includes a pine tree, a pine tree, or a pine tree. In the present specification, pine trees are classified into Pinus densiflora ), sea horse ( Pinus thunbergii or Pinus densiflora , and may be specifically harvested. In the present specification, the term "red pine" refers to a pine tree having a red epidermis, and the term "pine tree" means that the stem is straight and dark red, and the outline is similar to the sea.

In the method for continuously extracting essential oil and polyphenol from pine, which is one aspect of the present invention,

(c) extracting the polysaccharide from the polyphenol fraction.

According to another aspect of the present invention, there is provided a polyphenol obtained by the above method,

0.7% by weight or more of total flavonoids; And

Total proanthocyanidin may be a polyphenol containing 10% by weight or more.

The polyphenol which is one aspect of the present invention contains the above compound in a high content, so that the antioxidative ability can be maintained even though the polyphenol is obtained from the residue remaining after the oil separation. In view of the above, the polyphenols of the present invention may contain 0.9 wt% or more, 1.1 wt% or more, 1.3 wt% or more, 1.5 wt% or more, or 1.7 wt% or more of total flavonoids based on the total weight of polyphenols , And the total amount of proanthocyanidin may be 11 wt% or more, 12 wt% or more, 13 wt% or more, 14 wt% or more, or 15 wt% or more. In addition, the polyphenols of the present invention comprise less than 20% by weight of total flavonoids and less than 30% by weight of total proanthocyanidins.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are only for illustrating the present invention and that the scope of the present invention is not construed as being limited by these embodiments.

[Example 1] Pretreatment of pine samples

Leaves, twigs, and bark were harvested and cut into 20 mm. In the case of pine bark, finely grinded to less than 8 mesh resulted in a yield of more than several times in hydro distillation (oil yield: bark 0.03%, bark <8 mesh 0.07% / water layer yield: bark 1.5%, bark <8 mesh 3.12%) .

[Experimental Example 1] Extraction of essential oil and polyphenol

Essential oil and polyphenol were extracted by the same process as in Fig.

[Experimental Example 1-1] Steam distillation

In order to examine the possibility of industrial scale-up, the present inventors have devised and used a steam distillation apparatus of a scale capable of treating 1 kg or more of samples. For this purpose, a hole made in a stainless steel container was installed and a steam line was installed, and a line was drilled upward to allow the incoming steam to carry the essential oil.

After extracting the essential oil, the steam is condensed in a condenser installed on the way to the outlet and falls into the fractionation funnel. Steam was prepared by heating 8/10 of distilled water in a 5L plast, followed by heating in a heating mantle. The sample was placed in a stainless steel basket so that the hot steam produced could pass through the sample well. The liquid in the fractionation funnel passed through a pre-poured hexane layer (99.9% / 100 mL), where the nonpolar essential oil component remained in hexane.

The essential oil was separated from the obtained distillate, and the remaining material after distillation was used to obtain polyphenol. 1 kg of the residue was put into a 10 L capacity glass container, and 4 L of distilled water was added thereto to sufficiently immerse the residue, followed by hot extraction at 100 ° C. for 3 hours to extract the polyphenol component. The polyphenol was filtered and concentrated under reduced pressure and lyophilized to prepare a polyphenol fraction.

At this time, the polyphenol fraction can be extracted and the remaining residue can be used as a material capable of extracting a polysaccharide using a method such as hydrolysis enzyme or acid hydrolysis.

[Experimental Example 1-2] Hydro distillation

Continuous extraction of essential oil and polyphenol using the hydro-distillation method was also carried out as shown in Fig.

The pine samples of Example 1 (1 kg of leaves, 1 kg of twigs, 1 kg of stems) were placed in a round container and distilled water (3 to 4 L) was added to the sample to sufficiently immerse the sample and hydrolyzed for 6 hours. The hydro distillation unit is designed to boil the sample from a round vessel and to drop the essential oil down through the condenser at the top, where a pre-made hexane (100 mL) / water layer is available to collect the essential oil into a hexane layer . The hexane layer thus obtained is transferred to a container, and magnesium sulfate is added thereto, and then stored in a refrigerator for one day. The hexane layer was passed through filter paper, and then concentrated under reduced pressure at room temperature to remove the solvent to prepare pure water.

The essential oil was separated from the obtained distillate, and the remaining material after distillation was used to obtain polyphenol. 1 kg of the residue was put into a 10 L capacity glass container, and 4 L of distilled water was added thereto to sufficiently immerse the residue, followed by hot extraction at 100 ° C. for 3 hours to extract the polyphenol component. The polyphenol was filtered and concentrated under reduced pressure and lyophilized to prepare a polyphenol fraction.

The polyphenol fraction can be extracted and the remaining residue can be used as a material capable of extracting a polysaccharide using a method such as hydrolysis enzyme or acid hydrolysis.

[Example 3] Comparison of extraction ratio

As shown in FIG. 2, the hydrolysis rate of the crude oil was higher than that of the crude oil. However, it was not shown in this result, but it was confirmed that when extracting essential oil by hydro distillation after grinding bark under 8mesh, it was 0.07 higher than doubled. The water extracts showed no significant difference according to steam- and hydro distillation.

[Example 4] Comparison of antioxidant ability

(1) ABTS radical scavenging test

ABTS (2,2'-azino-bis (3-ethylbenzthiazoline-6-sulphonic acid)) reacts with potassium persulfate to form an unstable radical while losing electrons. The antioxidant scavenges this radical with the highest absorbance at 734 nm and can lose the harmfulness of the radical.

The sample equivalent (10 mg) of each polyphenol fraction extracted in each step of Example 2 was diluted in distilled water. After adding 5 ul of each diluted sample to 96-well, 95 ul of ethanol was added, and 100 ul of ABTS radical was added and reacted for 5 minutes. Thereafter, the radical scavenging ability was measured at 734 nm and expressed as%. On the other hand, as a control group for comparing antioxidant activity, pycnogenol, which is industrially used as a pine polyphenol fraction, was used.

As a result, the results shown in Fig. 3 were obtained.

(2) DPPH radical scavenging test (DPPH radical scavenging test)

DPPH (2,2-diphenyl-1-picrylhydrazyl) radicals dissolved in organic solvents have maximum absorbance at 515 nm. Antioxidants dissolve DPPH radicals and lose their intrinsic color and change their transparency.

For the measurement of DPPH radical scavenging, the sample equivalent (10 mg) of each of the pine polyphenol fractions extracted in the same manner as in Example 2 was diluted in distilled water. After adding 5 ul of each diluted sample into 96-well, 95 ul of ethanol was added, and 100 ul of DPPH radical was added and reacted for 30 minutes. Thereafter, the radical scavenging ability was measured at 515 nm, and the scavenging activity was expressed in%. On the other hand, pycnogenol, which is industrially used as a pine polyphenol fraction, was used as a control for comparison of antioxidant activity.

As a result, the results shown in Fig. 4 were obtained.

[Example 5] Comparison of contents of antioxidants

(1) Total flavonoid content measurement

The sample equivalent (10 mg) of each of the pine polyphenol fractions obtained in Example 2 was diluted in distilled water. 20ul of each diluted sample was put into a 96-well, and then 80ul of distilled water was added to dilute to 100ul. Here the addition of AlCl ₃ 6H ₂ O in 100ul reaction was carried out for 5 minutes. After that, the absorbance at 430 nm was measured, and the total flavonoid content was compared with the calibration curve of quercetin, which was a standard substance, and quercetin equivalent mg / g sample was measured. The positive control group was compared with the pine polyphenol fraction using pycnogenol.

As a result, the results shown in Fig. 5 were obtained.

(2) Determination of total proanthocyanidin content

The sample equivalent (10 mg) of each of the polyphenol fractions obtained in Example 2 was diluted in distilled water. The content of total proanthocyanidins was determined by the vanillin assay. That is, 2.5 mL of 1% vanillin solution and 2.5 mL of 25% (v / v) H2SO4 solution were added to 1 mL of each extract sample (1 mg / mL) and reacted at 25 ° C for 15 minutes. Thereafter, the absorbance of the reaction solution was measured at 500 nm using a spectrophotometer (Perkin-Elmer Lambda 15 UV-vis spectrophotometer, Nor-walk, CT). (+) - catechin (Sigma-Aldrich) was used as a standard substance and total proanthocyanidin content was expressed as CE ((+) - catechin equivalent) mg / g sample using a calibration curve of (+) - catechin Respectively.

As a result, the results shown in Fig. 6 were obtained.

(3) Total polyphenol content measurement

The sample equivalent (10 mg) of each of the pine polyphenol fractions obtained in Example 2 was diluted in distilled water. 20ul of each sample was placed in a 96-well and diluted to 80ul with 60ul of distilled water. To this was added 20 ul of Folin-Ciocaltene reagent and allowed to react for 5 minutes, then Na ₂ CO ₃ 100 ul was added. After 30 minutes, the absorbance at 730 nm was measured and the total polyphenol content was compared with gallic acid equivalent mg / g sample by substituting it into the calibration curve of gallic acid. As a positive control, pycnogenol was used and compared with the pine polyphenol fraction.

As a result, the results shown in Fig. 7 were obtained.

While the present invention has been particularly shown and described with reference to specific embodiments thereof, those skilled in the art will readily appreciate that many modifications are possible in the exemplary embodiments without materially departing from the novel teachings and advantages of this invention. something to do. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims (8)

delete (a) hydro-distilling the leaves of the pine tree to produce essential oil; And
(b) separating the essential oil from the obtained distillate, and subjecting the remaining material after distillation to hot water extraction to obtain a polyphenol containing proanthocyanidin,
The yields of the essential oil and the polyphenol can be obtained by processing the leaves of pine trees in the same manner as in the above steps (a) and (b) but using essential oil obtained by processing the leaves of pine trees by steam distillation instead of hydro distillation, , &Lt; / RTI &gt;
The content of proanthocyanidins can be determined by processing the leaves of pine trees in the same manner as in steps (a) and (b) above, but using proanthocyanidins obtained by processing pine leaves by steam distillation instead of hydro distillation Wherein the extraction yield of proanthocyanidin is increased by continuously extracting essential oils and polyphenols from pine trees.
[3] The method according to claim 2, wherein the pine leaves have a size of 8 mesh or less, and continuously extract essential oil and polyphenol from pine trees and increase extraction yield of proanthocyanidin.
3. The method of claim 2, wherein step (b)
A process for the processing of pine trees which comprises continuously extracting essential oils and polyphenols from pine trees and increasing the extraction yield of proanthocyanidins, comprising separating the essential oil with hexane.
delete The method according to claim 2, wherein the pine tree includes a pine tree, a pine tree, or a pine tree, which continuously extracts essential oil and polyphenol from the pine tree and increases the extraction yield of proanthocyanidin.
[3] The method according to claim 2,
(c) further comprising extracting the polysaccharide from the polyphenol fraction to continuously extract essential oils and polyphenols from the pine tree and increase the extraction yield of proanthocyanidin. delete

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