WO2013151407A1 - Method for extracting acorus calamus var. angustatus oil containing large amount of β-asarone using supercritical fluid extraction - Google Patents

Description

Extraction method of iris oil containing a large amount of beta asaron using supercritical fluid extraction

The present invention relates to a method for extracting iris root oil containing a large amount of beta asaron using a supercritical extraction method, more specifically, drying irises; Grinding the dried irises; The method relates to a method for extracting an iris oil containing a large amount of beta asaron from an iris comprising extracting the iris powder by a supercritical extraction method using supercritical carbon dioxide and a cosolvent and concentrating the iris extract. .

Iris ( Acorus calamus var. Angustatus ) is a perennial herb that grows in wetlands and belongs to Tiannan Province . The habitat is known to extend from Jeju Island to the northern part of Korea. In Korea, it is customary to bathe or wash your hair in water boiled with iris leaves or iris roots on a single day in a three-day period to defeat bad energy.If you wash your hair in water boiled with iris leaves or iris roots, your hair will not fall out. She believed that she was shiny and smooth, and her hair would be long like a window stalk.

In oriental medicine, it has important effects such as sedation, palliative action, analgesic action, and antifungal action. In addition, there is an aromatic substance, which is widely used as a fragrance for making lotion and high-quality perfume.

In addition, Korean studies have shown that iris oil has an inhibitory effect on the growth of Streptococous mutans , which is an oral cavity (You, YS, Park, KM and Kim, YB, Kor.J.Appl.Microbiol.Biotechnol, 21 (2), 187- 191 (1993)), the inhibitory effect of chicks and mice (Park, JH, Reserch review of Kyungpook Natl. Univ., 21 (1) 637-642 (1978)) and antioxidant efficacy (Hwang. MS, The theme of master of science in the Graduate of HEalth Sciences, Catholic Univ. of Daegu (2004).

Asarone of 4-propenyl-1,2,5-trimethoxybenzene (C ₁₂ H ₁₆ O ₃ , molecular weight 208.254 g / mol) is a major component of root, leaf and stem extracts of Acorus species. Korean iris species, which are native to Korea, are traditionally contained in large amounts of antibacterial components, and are known to exhibit antimicrobial and insecticidal effects such as treatment of hair dandruff and skin diseases (acne, athlete's foot, eczema, scabies, atopic bacteria, etc.) or food poisoning. Recently, it is used as a chemical sterilizer after steam treatment, and is known as a toxin, a morph modifier or an insect stimulant.

On the other hand, as isomer of asaron, beta-aron (β-asarone) is toxic while alpha-asarone (α-asarone) has no toxicity and has only anti-freezing activity. Asaron is known to be contained in a large amount of irises, among other plants, and irises are native to temperate regions and are aquatic plants native to any region including Jeju Island in Korea. Roots are beard roots, but long, long stems extend into the ground, and roots sprout from the bottom. Essential oils contain 0.8 to 5% in the root stem and 0.1 to 0.3% in the leaves, and beta asaron is one of the essential oil components. Pharmacological action is used as abortion, analgesic, aphrodisiac, fragrance, puncture, antiperspirant, tonic economy, antipyretic, hallucinogenic drug, homeopathic, sedative, stimulant, tonic or insect repellent.

Beta asaron extraction from conventional irises has been performed mostly through hot water extraction or methanol solvent extraction. As described above, the conventional extraction method has a problem in that residual waste and organic solvent remain after extraction. In addition, there is a disadvantage that the extraction yield of beta asaron is low.

Under this background, the inventors grind the irises to 0.5 mm to 1 mm particle size after natural drying and extract a large amount of beta from the irises by supercritical extraction using supercritical carbon dioxide and co-solvent. It was confirmed that the oil containing asaron can be extracted and completed the present invention.

It is an object of the present invention to provide a method for extracting iris oil containing beta asaron derived from irises using supercritical fluid extraction.

Another object of the present invention is to provide an iris oil containing beta asaron extracted using the above extraction method.

In order to solve the above problems, the present invention comprises the steps of crushing the iris (step 1); Supercritical extraction of the iris powders using supercritical carbon dioxide and co-solvent at 40 ° C. to 60 ° C. and 200 bar to 400 bar pressure conditions (step 2); And it provides a method for extracting beta-aron containing iris oil comprising the step of concentrating the iris extract (step 3).

Step 1 is a pretreatment method for increasing the extraction yield of the irises, the crushed irises to a predetermined size.

As used in the present invention, the term "Iris" is used as a medicinal herb sprayed with perennial herbaceous plants belonging to Chunnamseong. It is known to have the effects of dry stomach, soothing, jingyeong, and leech, and is used for symptoms such as digestion, diarrhea, wet swelling, jeongan, vigilance, forgetfulness, mental anxiety, customs pain, seawater, bronchitis, carbuncle, and spear. In addition, the root contains a fragrant substance that is used in the bath water.

It is preferable that the said iris uses the low temperature cold air drying dried iris. As used herein, the term "cold cold air drying" means a method of drying at low temperature. In the drying process, the low temperature means a relatively low temperature, and generally the low temperature means a temperature range of 40 ° C to 60 ° C. The low temperature cold wind drying in the present invention can be carried out for several days. It is preferably performed for 1 to 5 days, but may be appropriately adjusted according to the moisture content of the irises.

It is preferable to use a grinder, and the grinding is preferably a pin mill or a low temperature mill, but is not limited thereto.

In addition, the diameter of the iris powder is preferably a size of 0.5 mm to 1 mm. When the particle size of the pulverized product is out of the range, there is a disadvantage in that the extraction efficiency of the iris is lowered. In particular, the particle size of less than 0.5 mm, the extraction efficiency is lowered due to the pressure load during supercritical extraction, the beta asaron contained in the particles difficult to extract to the outside by the supercritical fluid at the particle size of more than 2 mm have.

Step 2 is a step of supercritical extraction by adding a co-solvent to the supercritical carbon dioxide in order to extract the iris oil containing beta asaron from the iris.

The cosolvent is preferably ethanol, but is not limited thereto. In addition, the cosolvent may use 1 to 10 vol% of the total supercritical fluid mixture, preferably 3 vol% of the total supercritical fluid mixture.

In addition, the supercritical extraction is preferably performed at a temperature of 40 ℃ to 60 ℃, it is preferably carried out at 200 bar to 400 bar pressure conditions.

Accordingly, as a result of the use of 0.5 mm particle size iris powder as an embodiment of the present invention, a high yield of iris oil supercritical extract of 6.2% was obtained under the extraction conditions of 50 ℃ and 200 bar, 40 ℃ and 300 It was confirmed that the iris supercritical fluid extract extracted by the addition of the co-solvent in the extraction condition of bar had a high content of beta asaron of 0.937 g.

Step 3 is a step of concentrating the iris supercritical extract to remove the cosolvent in the iris extract obtained by extraction. The concentration can be carried out using conventional thickeners known in the art.

In addition, the present invention provides a iris oil containing a large amount of beta asaron derived from irises extracted by the above extraction method.

The method for extracting beta-asarone derived from irises according to the present invention includes supercritical extraction of crushed irises having a particle size of 0.5 mm to 1 mm by adding a co-solvent to a carbon dioxide supercritical fluid, thereby containing a large amount of beta-asarone from irises. It is effective to provide a method for extracting essential oils.

FIG. 1 shows the addition of 2 ml / min ethanol at 40 ° C. of a 0.5 mm particle size iris mill according to one embodiment of the invention before (a) extraction, (b) after 200 bar supercritical extraction, and (c) Images show the iris extract after 300 bar supercritical extraction and (d) iris powder after extraction at 400 bar and (e) 200 bar extraction, (f) 300 bar extraction and (g) 400 bar extraction.

FIG. 2 shows the addition of 2 ml / min ethanol at 400 bar of a 2 mm particle size iris mill according to one embodiment of the invention before (a) extraction, (b) after 50 ° C. supercritical extraction and (c) An image showing the iris powder after 60 ° C supercritical extraction and (d) 50 ° C supercritical extraction and (e) 60 ° C supercritical extraction.

Figure 3 is (a) before extraction, (b) after 200 bar supercritical extraction, (c) after 300 bar supercritical extraction at 50 ° C of 0.5 mm particle size iris powder according to an embodiment of the present invention and ( d) An image showing the iris powder after extraction at 400 bar and (e) 200 bar extraction, (f) 300 bar extraction and (g) 400 bar extraction.

FIG. 4 shows the addition of 2 ml / min ethanol at 50 ° C. of a 0.5 mm particle size iris mill according to one embodiment of the invention before (a) extraction, (b) after 200 bar supercritical extraction, and (c) Images show the iris extract after 300 bar supercritical extraction and (d) iris powder after extraction at 400 bar and (e) 200 bar extraction, (f) 300 bar extraction and (g) 400 bar extraction.

Figure 5, (a) change in pressure, temperature change and extraction yield (%) change with or without cosolvent at 0.5 mm particle size according to an embodiment of the present invention, (b) temperature change at 200 bar pressure conditions And change in extraction yield (%) with and without cosolvent, (c) change in extraction yield (%) with temperature change at 300 bar pressure, and (d) temperature change and presence or absence of cosolvent at 400 bar pressure. It is a graph showing the change in extraction yield (%).

Figure 6 is a graph showing the results of HPLC quantification of (a) 7000 ppm, (b) 700 ppm and (c) 350 ppm for the beta asaron standard according to an embodiment of the present invention.

7 is an image showing a calibration curve measurement result obtained to verify the results of HPLC quantitative analysis by concentration of the beta asaron standard according to an embodiment of the present invention.

8 shows HPLC results of (a) extraction oil using co-solvent under 50 ° C. and 200 bar pressure conditions and (b) extraction oil using co-solvent under 40 ° C. and 300 bar pressure conditions according to an embodiment of the present invention. Is a graph.

9 is (a) extraction yield change of Example 4 and Example 13 and Comparative Example 1 of the high extraction yield according to an embodiment of the present invention; And (b) (b) extraction of beta-asarone total content in the iris oil according to the extraction conditions of Example 2, Example 13, and Comparative Example 1 of the high beta-aron content.

10 is a graph showing changes in total content of beta-aron and (b) change in total amount of beta-aron according to pressure change of (a) temperature change of an iris extract according to an embodiment of the present invention.

Figure 11, (a) the total content of the beta asaron and (b) the extraction yield (%) change according to the particle size and temperature change at 400 bar pressure condition of the iris supercritical extract according to an embodiment of the present invention The graph shown.

Hereinafter, the present invention will be described in more detail with reference to Examples. However, the following examples are only for illustrating the present invention and the scope of the present invention is not limited by these examples.

Examples 1 to 22: Iris supercritical extraction

The irises inhabiting Hapyeongsan, Korea were naturally dried at 60 ° C. for 5 days and then pretreated by grinding to a size of 0.5 mm to 2 mm using a pin mill. Supercritical extraction is controlled at temperatures of 40, 50 and 60 ° C., and the pressure is adjusted to 200, 300 and 400 bar at each temperature condition so that the amount of 100 g of the pretreated irises is extracted using a supercritical extraction device at each extraction condition. And supercritical extraction was performed for 100 minutes for each iris sample. In supercritical extraction, the flow rate was 60 ml / min (supercritical fluid extraction). In addition, when extracted using a combination of supercritical carbon dioxide and co-solvent, after maintaining the initial pressure, extracted with pure supercritical carbon dioxide for 1 minute and extracted with 2 ml / min ethanol (alcohol) for 60 minutes 39 Extraction was performed with pure supercritical carbon dioxide for minutes (SFE with co-solvent). When the supercritical extraction was completed, the pressure of the extraction tank of the supercritical extraction device was lowered to release the supercritical fluid state, and the extract was recovered and concentrated using a concentrator. Detailed extraction conditions are shown in Table 1 below, and images of iris powder and iris extract are shown in FIGS. 1 to 4.

Table 1 Example number Extraction condition Extraction temperature (℃) Extraction pressure (bar) Extraction time (min) co--solvent condition Particle size (mm) One SFE with co-solvent 40 200 100 2 ml / min, 60 min 0.5 2 SFE with co-solvent 40 300 100 2 ml / min, 60 min 0.5 3 SFE with co-solvent 40 400 100 2 ml / min, 60 min 0.5 4 SFE with co-solvent 50 200 100 2 ml / min, 60 min 0.5 5 SFE with co-solvent 50 300 100 2 ml / min, 60 min 0.5 6 SFE with co-solvent 50 400 100 2 ml / min, 60 min 0.5 7 SFE with co-solvent 60 200 100 2 ml / min, 60 min 0.5 8 SFE with co-solvent 60 300 100 2 ml / min, 60 min 0.5 9 SFE with co-solvent 60 400 100 2 ml / min, 60 min 0.5 10 SFE 40 200 100 - 0.5 11 SFE 40 300 100 - 0.5 12 SFE 40 400 100 - 0.5 13 SFE 50 200 100 - 0.5 14 SFE 50 300 100 - 0.5 15 SFE 50 400 100 - 0.5 16 SFE 60 200 100 - 0.5 17 SFE 60 300 100 - 0.5 18 SFE 60 400 100 - 0.5 19 SFE with co-solvent 50 400 100 2 ml / min, 60 min 2 20 SFE with co-solvent 60 400 100 2 ml / min, 60 min 2 21 SFE 50 400 100 - 2 22 SFE 60 400 100 - 2

Comparative Example 1: Iris methanol extraction

The irises inhabiting Hapyeongsan, Korea were dried at 60 ° C. for 5 days and then ground to a size of 0.5 mm using a grinder. 100 g of the prepared iris was put in 500 ml of a methanol solution, and the extract was supported for 24 hours at a temperature of 60 ° C., and then concentrated by using a vacuum concentrator to completely remove the solvent. Table 2 shows the extraction conditions.

TABLE 2 Comparative Example Number Extraction conditions Extraction temperature (℃) Extraction time (hr) One Methanol extraction 60 24

Experimental Example 1: Measurement of extraction yield according to temperature and pressure extraction conditions

The extraction yield of the iris extract obtained in the above Examples and Comparative Examples was measured. When the supercritical fluid extraction yield was measured, when the co-solvent was not used, 100 g (A) of the sample was put in the extraction basket, and the weight (B) of the extracted material obtained after the extraction was recorded.

Extraction Yield (%) = (B / A) * 100

Extraction yield measurement using cosolvent was carried out using supercritical carbon dioxide and cosolvent, supercritical extract material containing calamus oil was placed in a vacuum concentrator, and solvent was completely removed. 100 g (A ′) was recorded and calculated by the following equation.

Extraction Yield (%) = (B '/ A') * 100

The extraction yield obtained by the above method is shown in Table 3 below. As shown in Table 3, it was confirmed that the yield in the 40 to 50 ℃ temperature condition changes significantly. Therefore, it was confirmed that the temperature and pressure, the use of the co-solvent has a significant effect on the extraction yield and the content of the beta-saron of beta-aron containing beta-aron, the main influence factors I could see that.

TABLE 3 Example Number Extraction condition Extraction yield (%) One SFE with co-solvent 5.565 2 SFE with co-solvent 5.527 3 SFE with co-solvent 5.684 4 SFE with co-solvent 6.220 5 SFE with co-solvent 5.713 6 SFE with co-solvent 5.765 7 SFE with co-solvent 5.214 8 SFE with co-solvent 5.531 9 SFE with co-solvent 5.540 10 SFE 3.682 11 SFE 4.296 12 SFE 4.321 13 SFE 4.388 14 SFE 3.881 15 SFE 4.073 16 SFE 4.348 17 SFE 4.437 18 SFE 4.088 19 SFE with co-solvent 3.400 20 SFE with co-solvent 4.317 21 SFE 2.376 22 SFE 3.914 Comparative Example 1 methanol 3.253

The results of Table 3 are shown in FIG. 5 according to temperature and pressure conditions. As shown in Figure 5, it can be seen that the extraction yield of the sample prepared in Example 4 (0.5 mm particle size, 50 ℃ temperature and 200 bar pressure conditions) was the highest, each pressure condition (200 bar, 300 bar And (b), (c) and (d) of Fig. 5 showing the extraction yield change according to 400 bar) to confirm that the sample using the co-solvent extraction yield (%) is higher than the sample without the co-solvent Could.

In addition, in the supercritical fluid extraction results with the sample particle size of 0.5 mm, the highest yield of iris oil extraction was obtained at 50 ° C and 200 bar pressure, and the temperature was increased from 40 ° C to 50 ° C at the same pressure condition. The extraction yield increased at, whereas the yield decreased at 60 ℃. This is a predictable result of the correlation between the density of supercritical carbon dioxide and the density of the extracted iris oil. In particular, it was confirmed that the extraction yield was up to 2 times higher in the extraction conditions using the co-solvent than the extraction conditions using the supercritical carbon dioxide alone.

Experimental Example 2: Measurement of the iris supercritical extract component

HPLC analysis was performed using Waters HPLC with COSMOSIL 5C18MS-II, 250 mm × 4.6 mm (Nakala Teque, Inc., Japan) column at 10 μl injection volume and 0.1% formic acid (Fluka) as mobile phase. , USA) and acetonitrile (acetonitrile, JT Baker, USA) were eluted for 45 minutes at a flow rate of 1 ml / min using a concentration gradient (acetonitrile content: 20 wt% to 80 wt%). The oven temperature was set at 35 ° C. and the eluted material was detected using a PDA (Waters 2998, UV: 190 nm to 800 nm, Waters, USA) as a detector. Beta asaron purchased from Sigma Chemical Co., St Louis, MO was used as a standard.

1) Comparative measurement of components of irises supercritical extract and standard substances

The components of the irises supercritical extracts of Examples 1 to 22 were examined through HPLC analysis, and the results are shown in FIG. 9. As shown in FIG. 9, the extraction yield of the iris extract extracted through Example 4 was 1.9 times higher than that of the methanol extract obtained from Comparative Example 1, and the beta asaron content of the iris extract extracted through Example 2 was obtained. It was confirmed that 1.9 times higher compared with the control Comparative Example 1.

In addition, as a control for comparing the content of beta asaron content, the iris methanol extract extracted in Comparative Example 1 was used, to determine the concentration of beta asaron contained in iris oil, the sample by concentration using a beta asaron standard solution. The chromatograms prepared and obtained through HPLC analysis are shown in FIG. 6. The calibration curve for determination of concentration through the chromatogram obtained in FIG. 6 is shown in FIG. 7, and the conditions of the highest extraction yield and the high beta-asone content are shown in FIG. 8. As shown in Figure 8, the highest extraction yield was obtained under the conditions of Example 4 (50 ℃, 200 bar and co-solvent use conditions), the conditions of Example 2 (40 ℃, 300 bar and co-solvent use) Conditions) was found to be the highest concentration of beta-aron in the iris oil. In addition, as shown in Figure 9, the extraction conditions were significantly higher in Example 4 conditions (50 ℃, 200 bar and co-solvent use conditions) compared to Example 13 and the control Comparative Example 1 without the co-solvent, The conditions of Example 2 (40 ° C., 300 bar and cosolvent use conditions) were confirmed to be significantly higher in beta-asarone content in the iris oil than in Example 13 without the cosolvent and Comparative Example 1.

2) Determination of Beta-Asaron Content of Iris Extract

HPLC analysis was performed on the irises supercritical extracts obtained in Examples 1 to 22 to determine the content of beta asaron contained in each extract. On the other hand, the iris methanol extract extracted in Comparative Example 1 was used as a control for the beta asarone content fertilizer, the results are shown in Table 4. As shown in Table 4, it was confirmed that the beta-asarone content of the iris extract obtained in Example 2 was the highest.

Table 4 Example Number Beta Asaron Content (ppm) Total Beta-Asarone Content in Extracted Oil (g) One 631.321 0.703 2 847.517 0.937 3 592.073 0.389 4 663.120 0.825 5 650.636 0.743 6 622.146 0.717 7 585.448 0.611 8 617.495 0.683 9 651.493 0.722 10 726.000 0.535 11 684.073 0.588 12 633.367 0.547 13 592.244 0.520 14 691.393 0.537 15 667.558 0.544 16 768.869 0.669 17 589.138 0.523 18 773.870 0.633 19 564.918 0.269 20 560.337 0.339 21 763.327 0.254 22 725.935 0.398 Comparative Example 1 743.060 0.483

Changes in the total content of beta-asarone according to the temperature and pressure conditions of the iris supercritical extract using 0.5 mm particle size except 19 to 22 of Examples 1 to 22 are shown in FIG. 10. As shown in FIG. 10, the highest beta-aronone content was obtained in the extraction conditions using 40 ° C., 300 bar, and the cosolvent prepared in Example 2, and the sample using the cosolvent at the same temperature was used as the cosolvent. It was confirmed that the total extraction content of beta asaron is higher than the sample not used.

3) Component analysis of Iris supercritical extract under 400 bar pressure according to sample particle size

In Examples 1 to 22, the beta-aronon HPLC measurement concentration and the concentrated sample content of Examples 6, 9, 13, 18, 19, 20, 21, and 22 having a pressure condition of 400 bar were measured. And FIG. 11. As shown in Table 5 and Figure 11, regardless of the extraction temperature and pressure change, when using the particle size of 0.5 mm, the extraction yield was up to 2.4 times higher than that of the particle size of 2 mm, beta asaron The content was also confirmed to be as high as the extraction yield.

Cheongpo extract obtained in Example 6 showed the highest content of beta asaron, it was confirmed that the amount of extraction decreases slightly with increasing temperature. In addition, it was confirmed that the extraction amount increases as the co-solvent is present and the particle size is small. Therefore, it can be seen that the particle size and the use of the co-solvent is an important factor.

Table 5 Example number Condition Temperature (℃) Pressure (bar) Beta asarone measurement concentration (ppm) Concentrated Beta Asaron Content (g) Concentrated Sample Content (%) 6 SFE + co-solvent (0.5 mm) 50 400 622.146 0.717 5.765 19 SFE + co-solvent (2 mm) 50 400 564.918 0.269 2.38 13 SFE (0.5 mm) 50 400 667.558 0.520 4.073 21 SFE (2 mm) 50 400 763.327 0.254 1.663 9 SFE + co-solvent (0.5 mm) 60 400 651.493 0.722 5.54 20 SFE + co-solvent (2 mm) 60 400 560.337 0.339 3.022 18 SFE (0.5 mm) 60 400 773.870 0.633 4.088 22 SFE (2 mm) 60 400 725.935 0.398 2.74

Claims (5)

Crushing irises; Supercritical extraction of the iris powder into supercritical carbon dioxide using a co-solvent under 40 ° C. to 60 ° C. temperature and 200 bar to 400 bar pressure conditions; And Beta asaron extraction method comprising the step of concentrating the iris extract. The method of claim 1, wherein the diameter of the iris powder is beta asaron extraction method characterized in that the size of 0.5 mm to 1 mm. The method of claim 1, wherein the co-solvent is beta asaron extraction method characterized in that the ethanol. 5. The method of claim 4, wherein the cosolvent is 1 to 10 vol% of the total supercritical fluid mixture. The method of claim 1, wherein the iris is a dry iris.

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