KR102141609B1 - Manufacturing method for onion juice - Google Patents

Abstract

The present invention relates to a method for manufacturing onion juice comprising: a step of separating skin after washing an onion; a step of generating a ripening liquid by ripening onion skin separated in the step with a solvent; a step of breaking a raw onion from which the skin is removed into small pieces; a step of generating a mixing liquid by mixing the ripening liquid of the step and the raw onion processed in the step; a step of generating an active ingredient extracting liquid by adding the solvent to the mixing liquid of the step; and a step of removing the solvent from the active ingredient extracting liquid of the step. The present invention can increase an anticancer effect of the onion juice.

Description

Method of manufacturing onion juice {MANUFACTURING METHOD FOR ONION JUICE}

The present invention relates to a method for producing onion juice, and more particularly, to a method for producing onion juice with a higher quercetin content.

Onions have been used in various foods since ancient times as a healthy vegetable. Onions are mainly used for edible stems, and the peculiar smell from scale stems is due to compounds such as propyl disulfide and allyl sulfide. This physiologically promotes the secretion of digestive fluids and has the effect of excitement and diuresis. In addition, the scale stem contains various vitamins and minerals such as calcium phosphate, which has the effect of removing harmful substances from the blood. Scale stems are often used in salads, soups, and meat dishes, and as spices in various dishes. The leaves contain 5,000 IU of vitamin A, 45 mg of vitamin C, 80 mg of calcium, 24 mg of magnesium, and 220 mg of potassium in 100 g.

In particular, the effect of onion is that it clears the blood. This is the efficacy of quercetin, one of the active ingredients in onions.

Quercetin is a kind of flavonoid and widely distributed in plants. It is known to have cancer prevention, anti-inflammatory, diabetes improvement and complications prevention, nicotine detoxification, skin cancer prevention, and anti-viral action in the human body. Play a role.

Quercetin, which is abundant in onion, has good absorption and metabolism, and thus maintains a stable plasma concentration, so it is effective in preventing diseases of the vascular system and preventing progress.

In addition, the control of blood sugar and the supply of antioxidants to diabetic patients are essential for the prevention and sedation of complications caused by diabetes. As mentioned above, onions contain a lot of compounds that lower blood sugar and quercetin, an antioxidant, so people with diabetes Has been a great help.

Therefore, taking advantage of these onions, onion juice in the form of a drink is presented in various products so that onions can be easily consumed.

For example, Republic of Korea Patent Publication No. 10-2012-0077732 discloses a method of manufacturing onion juice containing a pig skin and onion juice prepared by the above method.

However, in producing or manufacturing onion juice in various ways, it is a key to include as much quercetin as possible, which has an excellent effect, as described above, and thus it is necessary to maximize the efficiency of quercetin extraction in onion juice production.

Registered Patent No. 10-1791206

The present invention has been devised to meet the above-mentioned conventional requests, and its object is to provide a method for preparing onion juice containing as much quercetin as possible from the same amount of onions.

In order to achieve the above object, a method for preparing onion juice according to the present invention includes washing the onion and separating the peel; A step of aging the onion peel separated in the step using a solvent to produce a aging liquid; Crushing the peeled raw onion into small sizes; Mixing the aged liquid of the step and the raw onion processed in the step to produce a mixed liquid; Generating an active ingredient extract by adding a solvent to the mixed solution of the step; It may include the step of removing the solvent from the active ingredient extract of the step.

Here, the solvent of the step may be ethanol.

Here, the separated onion peel may be aged using 65% to 75% ethanol to produce a aging liquid.

Here, ethanol can be removed.

Here, 45% to 55% ethanol may be added to the mixed solution to generate an active ingredient extract.

Here, the aging period is 18 months to 24 months, and may be aged at a low temperature of 7°C to 12°C or less.

Here, the aging solution can remove ethanol at a temperature of 80°C to 85°C, and the active ingredient extract can remove ethanol at a temperature of 80°C to 85°C.

Here, the aging liquid of the above step may be concentrated under reduced pressure at a temperature of 70°C to 75°C and a pressure of 40 cmHg to have a concentration of 10 to 15 Brix.

Here, the active ingredient extract may be concentrated under reduced pressure at a temperature of 70°C to 75°C and a pressure of 40 cmHg to have a concentration of 20 to 25 Brix.

As described above, according to the present invention, the anti-cancer effect of onion juice can be enhanced by maximizing the extraction amount of an active substance containing quercetin or the like.

1 is an overall process of a method for preparing onion juice according to an embodiment of the present invention,
2 to 5 is a view showing the anti-cancer effect between the onion juice and the other onion juice produced through the process according to FIG.

Hereinafter, the present invention will be described in detail with reference to the accompanying drawings.

Hereinafter, each embodiment according to the present invention is only one example to help understanding of the present invention, and the present invention is not limited to these embodiments. In particular, the present invention may be composed of a combination of at least one or more of the individual steps included in each embodiment.

In particular, for convenience, some claims in the claims include alphabets such as'(a)', but these alphabets do not define the order of each step.

The overall process for preparing onion juice according to an embodiment of the present invention is as shown in FIG. 1.

As shown in the drawing, the method for preparing onion juice according to an embodiment of the present invention largely separates the onion peel, the step of aging the separated onion peel, and breaks or crushes the peeled fresh onion into small sizes. The step, mixing the aging liquid using onion peels and crushed or crushed green onions in a small size to produce a mixed solution, extracting the active ingredient from the mixed solution, and removing the solvent from the active ingredient extract Can.

The explanation for each step is as follows.

After washing the onion, the skin is separated, and the separated onion skin is aged, where the onion skin is aged after mixing a predetermined solvent.

For example, it can be aged while allowing the active ingredient to be extracted using an ethanol extraction method, which is superior in effect to hot water extraction. For example, the peel can be aged using 68% to 75% ethanol to produce a aging liquid.

As such, the biggest feature of the present invention is that it undergoes a process of separately separating onion skins and ripening.

The aging period for the onion peel is 18 months to 24 months, and it is preferable to perform low temperature aging at 5°C to 10°C or less.

Through this aging period, many active substances (active ingredients) including quercetin contained in the onion peel are released.

After the aging period, a step for removing a solvent for aging, for example, ethanol, may be performed.

For example, the aging liquid can be removed by heating the medium at a temperature of 80° C. to 85° C. over medium heat.

Since the boiling point of ethanol is about 78°C under 1 atmosphere, most of the ethanol can be removed even at this temperature.

As another example, the onion peel ripening solution may be concentrated under reduced pressure at a temperature of 40° C. to 75° C. under a pressure of 40 cm Hg, wherein the concentration of the ripening solution is preferably 10 to 15 Brix.

On the other hand, the peeled onions are separately crushed or crushed, and the pulverizing method of the fruit or vegetable itself corresponds to a known technique, so a detailed description thereof will be omitted.

The pulverized or crushed raw onion is mixed with the onion peel ripening solution (preferably, the ethanol-removed ripening solution) to form a mixed solution.

That is, the mixed solution will contain the ripening solution produced by ripening the onion peel (of course, the ripening solution contains various active ingredients including quercetin) and fresh onion.

An active ingredient extract is generated by adding a solvent to the mixture for secondary extraction of the active ingredient.

For example, an active ingredient can be extracted using an ethanol extraction method that is superior in effect to hot water extraction. For example, an active ingredient extract may be generated by adding 45% to 55% ethanol to a mixed solution.

After the active ingredient extract is generated, a process of removing the previously added solvent, for example, ethanol, is performed. For example, the active ingredient extract may be heated to a temperature of 80°C to 85°C to remove ethanol.

As another example, the active ingredient extract may be concentrated under reduced pressure at a pressure of 40 cmHg to a temperature of 70° C. to 75° C. At this time, the concentration is preferably 20 to 25 Brix.

Hereinafter, specific examples of the method for preparing onion juice according to the present invention and onion juice according to the method and a diluent and control (onion juice prepared in different ways) and dilutions thereof will be compared.

(Example 1)

After adding 100 g of onion peel to 500 g of 70% ethanol, low temperature aging at 10° C. for 18 months to produce a aging liquid, and then concentrating under reduced pressure at a temperature of 73° C. and a pressure of 40 cm Hg to achieve a concentration of 13 Brix, peeled raw onion After crushing 1 kg (5 to 6 portions), mix with the aging solution, and 1 kg of 50% ethanol at room temperature to the mixed solution, wait 5 hours to become an active ingredient extract, and the active ingredient extract is 73 under a pressure of 40 cmHg. Concentration under reduced pressure at a temperature of ℃ to a concentration of 23 Brix.

The anticancer efficacy between these examples and onion juice having the same concentration as produced and sold by a third party and a comparison group was measured, and the process and results are as follows.

1. Cancer cell proliferation inhibitory effect measurement

1) Test method: MTT assay method Exclusion analysis method

2) Cell lines: Cancer cell lines such as colon cancer cells, lung cancer cells, and liver cancer cells are used.

3) Test sample: Onion juice according to the present invention, commercial onion juice analysis of two other companies, comparison.

4) Dose step: Adjust the onion juice solution according to the present invention to treat the concentration step by step with a 2x concentrated solution, a stock solution, a 5x diluted solution, or a 10x diluted solution.

2. Test process

1) Cell line culture

(1) Cell line culture:

Culture each cancer cell line in RPMI-1640 or DMEM medium containing 10% fetal bovine serum

(2) Cell line management: passage culture using trypsin-EDTA solution every 2-3 days

Management by conduct.

2) Cell viability measurement

(1) Cell division: RPMI-1640 or DMEM medium was added to the cultured cells.

Dispense 1 ml of cells into 24 well plates at 2 ㅧ 105 cells/ml.

(2) After cell dispensing, stabilize for about 1 hour, and then add 10ul of each sample for each concentration.

Add.

(3) After culture for 24 hours or 48 hours, 10ul of MTT (5mg/ml in PBS) was added to each well.

adding

(4) After incubation for 4 hours, formazan crystals generated were dissolved with DMSO and measured by measuring absorbance at 595 nm with an ELISA plate reader.

3) Cell survival and cell death measurement

(1) Cells cultured for 4-5 days were inoculated into culture dishes in the same amount at a density of 2ㅧ105 cells/dish, and cultured for one day for stabilization.

(2) After the onion juice was diluted step by step, cultured for an additional 2 days.

(3) Wash with PBS and harvest using trypsin-EDTA.

As a result of testing through the above-described process, results as shown in FIGS. 2 to 5 were obtained.

2 shows the viability of colon cancer cell A (% of control), and it can be seen that colon cancer cell A had a significantly lower survival rate in concentrate, stock solution, and dilution of onion juice according to the present invention than onion juice of other companies.

Figure 3 shows the colorectal cancer cell B viability (% of control), it can be seen that the colorectal cancer cell B survival rate is significantly lower in the concentrate, stock solution, dilution of onion juice according to the present invention than the other company's onion juice, 2 times In the concentrate, it can be seen that the survival rate of colorectal cancer cells B is lower.

Figure 4 shows the lung cancer cell A viability (% of control), it can be seen that the lung cancer cell A survival rate is significantly lower in the concentrate, stock solution, dilution of onion juice according to the present invention than onion juice of other companies.

In addition, Figure 5 shows the liver cancer cell viability (% of control), it can be seen that the liver cancer cell survival rate is significantly lower in the concentrate, stock solution, dilution of onion juice according to the present invention than the onion juice of other products.

Test results: Compared to onion juice and other onion juice products according to the present invention, the cancer efficacy was tested by applying to two types of colorectal cancer cell lines A and B, liver cancer cell line, and lung cancer cell line. 10 times dilution, 5 times diluted, undiluted solution, 2 times concentrated, concentrated to each concentration, treated to each cell line, incubated for 24 hours, and then treated with MTT solution to perform cell viability measurement, and the anticancer properties of onion juice according to the present invention The colon cell lines A and B, the liver cancer cell line, and the lung cancer cell line were all clearly shown, and the most pronounced anti-cancer properties when applied under the same conditions as the test method at the concentration of the stock solution.

When compared to the products of other companies' two places, the concentrate of the other company's products was applied twice as concentrated, and it was somewhat similar to the anticancer efficacy of the onion juice stock solution according to the present invention, but showed lower anticancer capacity than the product of the onion juice according to the present invention. .

Hereinafter, other embodiments compared with the above-described embodiment 1 will be compared.

(Comparative Example 1)

The same procedure as in Example 1 was carried out, so that the low-temperature aging period for the onion peel was set to 24 hours.

(Comparative Example 2)

It was carried out in the same manner as in Example 1, when preparing the ripening solution of the onion peel was used 500g of 50% ethanol.

(Comparative Example 3)

It was carried out in the same manner as in Example 1, but instead of concentrating the aging solution under reduced pressure, it was heated to a temperature of 80°C at 1 atmosphere so that the concentration was 13 Brix, and the extract of the active ingredient was also heated to a temperature of 80°C at 1 atmosphere instead of concentration under reduced pressure. To make 23Brix

(Comparative Example 4)

The same procedure as in Example 1 was carried out, but the process of concentration under reduced pressure in the aged liquid was omitted.

.

(Comparative Example 5)

Instead of using a third-party product (onion juice), it was carried out in the same manner as in Example 1, but crushed 1 kg of raw onion (5 to 6 portions) without peeling onion (i.e., ripening period) and then at room temperature. 1 kg of ethanol was administered to wait 5 hours to be an active ingredient extract, and the active ingredient extract was concentrated under reduced pressure at a temperature of 73° C. under a pressure of 40 cmHg to obtain a concentration of 23 Brix.

The results of the anticancer cell test described above with respect to these comparative examples and examples are shown in FIGS. 6 to 9.

When the results of the comparison (Example 1) were compared with other comparative examples, the anti-cancer effect at the cell level was confirmed in the test for each anti-cancer cell.

In other words, by separating the onion peels for a long period of low temperature aging, and also using an alcohol extraction method, the alcohol concentration for the onion peel is higher than the alcohol concentration for the onion from which the peel has been removed, and in particular, alcohol is used by using reduced pressure extraction. It can be seen that the removed case shows a better effect in the anticancer test.

In this case, when the onion peel is separated and aged at a low temperature, the alcohol extraction method for the onion peel uses a high concentration of alcohol different from the alcohol extraction method for onion, and also uses the alcohol extraction method to lower the pressure and lower the pressure rather than at room temperature. In the case of processing in the phosphorus state, the case where the decompression concentration process for the ripening solution of the onion peel was first performed may be interpreted as releasing a large amount of active substances for the death of cancer cells.

In particular, FIG. 10 shows the results of comparing the concentrations of quercetin among active ingredients extracted from onions.

The measurement of the quercetin concentration contained in each of the extracts prepared by the previous examples and each comparative group and comparative example was measured using a HPLC method (High Performance Liquid Chromatography method).

In order to perform the HPLC method, pretreatment was performed for each extract prepared in each example.

For example, pre-treatment was performed by adding about 50 mL of the active ingredient extract prepared in Example 1 into a 300 mL volume separation filter, adding the same amount of ethyl acetate, shaking and standing at 300 rpm for 5 minutes to separate the phases.

After phase separation, the lower aqueous layer was decanted with another separatory column, the upper ethyl acetate layer was transferred to an evaporate flask, and the aqueous layer separated by separatory distillation was repeated such that the method was three times in total.

The ethyl acetate layer combined by this repeating process was concentrated under reduced pressure in a 55° C. water bath to evaporate the solvent completely to obtain a residue, and 4 ml of methanol was completely dissolved in the obtained residue, washed completely, and then used a pipette (Pasteur pipette). After transferring to a 10 mL size flask and washing the flask again with methanol, the process of combining the washing solutions was repeated three times in total. Finally, filtering was performed with a 13 mm PVDF syringe filter (Whatman, 0.45 µm) and used as an HPLC sample.

HPLC system for measuring quercetin concentration (YOUNGLIN INSTRUMENT, Korea) includes: Solvent Delivery Pump (Model M930D); Absorbance Detector (Model M730D); It consists of Column Oven (Model CTS30) and Solvent Degassor & Valve Module (Model SDB30 plus), and the detection of quercetin separated by an HPLC system was performed at a measurement wavelength of 370 nm using an ultraviolet absorption photometer.

For the analysis using HPLC, the analysis column used ODSHYPERSIL (Thermo ELECTRON CO., Model 30105-154630), and as a specific specification, it was filled with 5 μm C18 in a stainless steel pipe having an analysis inside diameter of about 5 mm and a length of about 15 cm, and the column was The temperature was set to room temperature, and the mobile phase used a solution in which 0.04% of TFA (trifloroacetic acid) was added to distilled water (Milli-Q water) and methanol mixture (60:40), and the solvent flow rate of the mobile phase was 1 mL. /min, and the moving time was set to 30 minutes. In addition, the injection volumn of the sample for performing the HPLC was 10 ul.

The result of HPLC analysis of the quercetin standard material measured by this method is shown in FIG. 10.

Quercetin dehydrate (minimum 98% HPLC, SIGMA, USA) was used as a standard quercetin standard for measuring quercetin concentration.

More specifically, by performing HPLC analysis at the concentrations of the standard quercetins 50, 100, 200, 5000, 10000 and 20000 ppm, a standard curve was prepared by a linear equation (y = ax + b).

The measurement and calculation of the quercetin concentration (content) of the extracts prepared in the Examples is carried out by HPLC analysis of the extract as a sample, and the quercetin peak area measured by the analysis results is known formula. Calculated by substituting for (Quercetin concentration (ppm) = (a ㅧ Quercetin region + b) 여기서 10 (dilution multiple)), where the value multiplied by the formula means the dilution factor of methanol (10 times) during the pretreatment of the sample. .

As shown in FIG. 10, it can be seen that the quercetin concentration extracted from the same amount of onion is the case of Example 1 of the present invention, and it is determined that the anticancer effect described above was exhibited according to the increase in quercetin extract amount.

The present invention is not limited to the specific embodiments described above, but can be implemented by various modifications and modifications without departing from the gist of the present invention. It will be apparent that such variations and modifications are included in the present invention if they fall within the scope of the appended claims.

Claims (7)

(A) after washing the onion, separating the peel;
(B) a step of aging the onion peel separated in the step (a) using a solvent to produce a aging solution;
(c) crushing the peeled raw onion into small sizes;
(d) mixing the aged liquid of the step (b) with the raw onion processed in the step (c) to produce a mixed liquid;
(e) generating an active ingredient extract by adding a solvent to the mixed solution of step (d);
(f) removing the solvent from the active ingredient extract of step (e),
The solvent of the step (b) and the solvent of the step (e) are ethanol, and in the step (b), the separated onion peel is aged using 65% to 75% ethanol to generate a aging solution, and the (e) In step ), an active ingredient extract is generated by adding 45% to 55% ethanol to the mixed solution of step (d),
(g) after step (b), further comprising the step of removing ethanol in step (b),
The ripening period of step (b) is 18 months to 24 months, and onion juice production method characterized in that it is aged at a low temperature of 7°C to 12°C. delete delete delete delete According to claim 1,
In step (g), the aging liquid of step (b) is removed from ethanol at a temperature of 80°C to 85°C,
In the step (f), the method for producing onion juice, characterized in that it comprises removing the ethanol at a temperature of 80 ℃ ~ 85 ℃ the active ingredient extract of the step (e). According to claim 1,
In the step (g), the method of producing onion juice, characterized in that the concentration of the aging liquid of the step (b) is concentrated under reduced pressure at a temperature of 70°C to 75°C and a pressure of 40 cmHg so as to have a concentration of 10 to 15 Brix.
In the step (f), the method for producing onion juice, characterized in that the concentration of the active ingredient extract of step (e) is concentrated under reduced pressure at a temperature of 70° C. to 75° C. and a pressure of 40 cmHg so as to have a concentration of 20 to 25 Brix.

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