KR101032459B1 - Method for preparing extract of high quality chlorophyll - Google Patents

Abstract

PURPOSE: A method for extract chlorophyll extract from Camellia sinensis O. Kuntze leaf is provided to prevent discoloration and to maintain green color for a long time. CONSTITUTION: A method for preparing chlorophyll extract of high quality comprises: a step of pulverizing Camellia sinensis O. Kuntze leaf; a step of adding alcohol and mixing; a step of leaching for 9-14 hours and filtering; a step of adjusting the filtered liquid with basic solution by pH 10.0-12.0 and performing saponification at 50-70°C for 0.5-2 hours; a step of mixing the alcohol solution and petroleum ether in a volume ratio of 1-2:1-2 and removing a petroleum ether layer; a step of adjusting the solution by pH 3.0-5.0; a step of adding 5-10 weight% of alcohol solution and substituting at 50-70°C for 10-60 minutes.

Description

Method for preparing high quality chlorophyll extract {Method for preparing extract of high quality chlorophyll}

The present invention relates to a method for producing high quality chlorophyll extract from tea leaves.

Chlorophyll is a compound in the leaves of green plants and is contained in the chlorophyll's grana. There are several known chlorophylls such as a, b, c, d, e and bacteriochlorophylls a and b. Chlorophyll is used to assimilate carbon dioxide into carbohydrates, an organic compound, by absorbing light energy in the chloroplasts of green plants.

In recent years, as chlorophyll is spotlighted as a substance having health functionalities, intake and use are increasing. Representative physiological activities of chlorophyll include hematopoietic action, vitamin provision, enzymatic activity maintenance, detoxification, deodorization, fibrin provision, anti-inflammatory action, and metabolism promotion. That is, iron, which is a trace element contained in a large amount in chlorophyll, can be used as a natural hematopoietic raw material, and vitamin C and inorganic salts are also contained in chlorophyll in large amounts, thereby improving homeostasis of bioactivity. In addition, chlorophyll plays an important role in the production and maintenance of enzymes in the body, prevents infection, prevents the spread of inflammation, suppresses pain, and also removes various body odors generated in the body. In addition, since plant fiber is present together with chlorophyll, the ingestion of chlorophyll allows sufficient intake of fiber. In addition, it is known to improve the constitution and prevent aging, cancer, genetic mutations, gastrointestinal diseases, acne, blemishes, freckles, spots, anemia, chronic fatigue, heart disease, high blood pressure, stroke, hepatitis and cirrhosis.

However, despite the chlorophyll's potent efficacy, it has never been the only way to consume it, either by raw food or by directly eating green foods such as green tea, kale or mulberry leaves. That is, the most important problem in the application of chlorophyll is not the chlorophyll itself but its extraction method and concentration. Since the chlorophyll's structure and potential have been identified, the amazing effects of chlorophyll have been demonstrated in various studies and clinical trials for over 100 years, but the development of extraction methods has been insufficient. Japanese Patent Laid-Open Publication No. 59-33231 and Japanese Patent Laid-Open No. 2-83329 disclose a chlorophyll extracting device for extracting chlorophyll by ultrasonic wave in alcohol solution and the preparation of chlorophyll preparation, but exist in the extract by ultrasonic wave. Chlorophyll is a problem that is destroyed. In addition, a fatal problem that chlorophyll is destroyed because the extraction solvent of the extracted chlorophyll quickly evaporates to protect the chlorophyll contained in the extract.

The present inventors studied a method of separating chlorophyll with high health functionalities from the leaves of Camellia sinensis O. Kuntze and using them as food, and through various studies, the chlorophyll quality is preserved as it is. The present invention was completed by devising a method for producing high-quality chlorophyll extracts with high functionality.

An object of the present invention is to provide a method for producing a high quality chlorophyll extract.

Chlorophyll extract of the present invention,

(Step 1) grinding the dried tea leaves;

(Step 2) mixing the tea leaves and alcohol and leaching for 9-14 hours and then filtering;

(Step 3) adjusting the filtrate of step 2 with a basic solution to pH 10.0 ~ 12.0, and saponification reaction at 50 ~ 70 ℃ for 0.5 ~ 2 hours;

(Step 4) Take only the upper alcohol solution of the saponification reaction solution, mix the upper alcohol solution with petroleum ether in a volume ratio of 1-2: 1-2, remove the petroleum ether layer to remove pigments other than chlorophyll. step;

(Step 5) removing the petroleum ether layer from the mixture of step 4 and taking only the remaining alcohol solution, adjusting the pH to 3.0 to 5.0 with an acidic solution;

(Step 6) 5 to 10% by weight of an aqueous copper sulfate solution or zinc acetate solution to the alcohol solution of the pH of the step 5 is adjusted to 3.0 ~ 5.0 to replace the alcohol solution for 20 to 40 minutes at 50 ~ 70 ℃ step; And

(7 step) drying the substituent under reduced pressure to prepare a dried product, filtering the precipitated inorganic salt by dissolving the dried product in alcohol, and then drying the filtered alcohol solution under reduced pressure again;

Through the final chlorophyll extract is prepared.

Filtration of the tea leaf leachate in the said 2 step can be filtered using a filter paper, a cotton, a nonwoven fabric.

The alcohol used in the second step may be used 80 to 100% ethanol aqueous solution or 80 to 100% methanol aqueous solution.

In the third step, the leachate leaves a pigment material such as chlorophyll or carotene in the ethanol solution through a basic reaction and a saponification reaction, and other impurities are precipitated at the bottom.

In the step 3, the pH of the leaching solution may be adjusted using sodium hydroxide aqueous solution, potassium hydroxide aqueous solution, and the like, and the concentration of the aqueous solution is appropriately 10-20%.

Removing the pigment other than chlorophyll using the mixing method of the alcohol solution and petroleum ether of the four step may be repeated 2 to 4 or more times.

In the fifth step, the pH can be adjusted with concentrated hydrochloric acid, concentrated sulfuric acid, and the like.

The concentration of the copper sulfate aqueous solution or zinc acetate aqueous solution in the step 6 is suitably 3 to 7%.

Drying under reduced pressure in step 7 is carried out in a conventional manner, and the drying temperature is appropriately 40 ~ 60 ℃.

The drying step of step 7 may be repeated 2 to 4 or more times.

Inorganic salt precipitates such as sodium chloride or magnesium sulfate are precipitated through the drying step of the seventh step, so that the inorganic salt precipitates can be easily removed.

The chlorophyll extract prepared in step 7 may be dissolved in an edible oil such as soybean oil in some cases and refrigerated for use as a food additive.

Chlorophyll extract of the present invention provides a foodstuff comprising a food supplement acceptable food supplement. Food containing chlorophyll extract of the present invention includes tablets, capsules, pills or liquid form, such foods, such as drinks, meat, sausages, bread, candy, snacks, dairy products, soups, including ice cream, soup , Beverages including ionic beverages, alcoholic beverages and nutritional products including vitamin complexes may be included.

The present invention relates to a method for extracting chlorophyll from the tea leaves, through the chlorophyll extraction method of the present invention, it is possible to extract a large amount of high-quality chlorophyll of high purity. In addition, the color of the extracted chlorophyll can be maintained in green for a long time without discoloration.

1 is a flow chart showing a method for producing chlorophyll extract according to the method of Example 1 of the present invention.

Hereinafter, preferred embodiments of the present invention will be described in detail. However, the present invention is not limited to the embodiments described herein and may be embodied in other forms. Rather, the embodiments disclosed herein are provided so that the disclosure can be thorough and complete, and will fully convey the scope of the invention to those skilled in the art.

<Example 1. Extraction of Chlorophyll from Tea Leaves-①>

The dried tea leaves processed by the 10g thickening process were collected, crushed into fine triangle platters, put in 100 ml of 98% ethanol aqueous solution, leached in the dark for 12 hours, and filtered to remove the tea leaves. 45 ml of the filtrate was taken, and the pH was adjusted to 11.0 with 15% aqueous sodium hydroxide (NaOH) solution, followed by saponification reaction at 60 ° C for 1 hour. After saponification, the same amount of petroleum ether was added to the upper ethanol solution to remove pigments other than chlorophyll (chlorophyll, carotene, etc.). [Soapsing reaction: A precipitate forms at the bottom and chlorophyll can dissolve the non-chlorophyll pigments in the upper layer of ethanol / petroleum ether: ethanol solution. Layer separation occurs to facilitate separation of non-chlorophyll pigments (chlorophyll, carotene, etc.)]

After repeating the method of separating the pigments other than chlorophyll three times using petroleum ether as described above, the pH was adjusted to 4.0 using concentrated hydrochloric acid (HCl), and 2 ml of 5% copper sulfate solution was added to 30 ℃ at 30 ℃ Substitution reaction was performed for minutes.

The replacement reaction product was dried under reduced pressure in a conventional manner, and then dissolved by dissolving again in 10 ml of 98% ethanol to precipitate an inorganic salt precipitate. The filtered ethanol solution was dried under reduced pressure to obtain chlorophyll dried product.

<Example 2. Extraction of Chlorophyll from Tea Leaves-②>

Chlorophyll was extracted from the tea leaves in the same manner as in Example 1, but the tea leaves were leached with an 80% ethanol aqueous solution instead of the 98% ethanol aqueous solution.

<Example 3. Extraction of Chlorophyll from Tea Leaves-③>

Chlorophyll was extracted from the tea leaves in the same manner as in Example 1, but the tea leaves were leached with a 70% ethanol aqueous solution instead of the 98% ethanol aqueous solution.

Example 4 Extraction of Chlorophyll from Tea Leaves

Chlorophyll was extracted from the tea leaves in the same manner as in Example 1, but 2 ml of 5% zinc acetate aqueous solution was used instead of 2 ml of 5% copper sulfate aqueous solution.

Comparative Example 1. Extraction of Chlorophyll without 5% Copper Sulfate Solution

Chlorophyll was extracted from the tea leaves in the same manner as in Example 1, but the chlorophyll extract was prepared without performing the step of using a 5% copper sulfate solution.

Comparative Example 2. Extraction of Chlorophyll without pH Adjustment Step-③

Chlorophyll was extracted from the tea leaves in the same manner as in Example 1, but the chlorophyll extract was prepared without performing the step of adjusting the pH to 11 and the step of adjusting the pH to 4.

Comparative Example 3. Preparation of Chlorophyll Extract Using Ethanol Only

10 g of dried tea leaves were finely pulverized, placed in 100 ml of an aqueous 98% ethanol solution, and leached for 12 hours. The leachate was distilled under reduced pressure to prepare a chlorophyll extract.

Comparative Example 4 Preparation of Chlorophyll Extract Using 30% Ethanol Solution

Chlorophyll was extracted from the tea leaves in the same manner as in Example 1, but 30% ethanol aqueous solution was used instead of 98% ethanol aqueous solution.

Comparative Example 5. Preparation of Chlorophyll Extract Using 50% Ethanol Solution

Chlorophyll was extracted from the tea leaves in the same manner as in Example 1, but 50% ethanol solution was used instead of 98% ethanol solution.

Experimental Example 1. Comparison of Extraction Efficiency and Purity of Chlorophyll Extracts

Chlorophyll extract prepared in Examples and Comparative Examples was confirmed by HPLC (high pressure liquid chromatography) how much chlorophyll is contained. HPLC was performed using an agiline 1200 system, with a sample injection volume of 10 μl and solvent flow rate of 1 mL / min. As a developing solution, a mixture of methanol and acetonitrile in a 1: 1 ratio was used.

The HPLC results are shown in Table 1 below, and show the degree of purification (purity of chlorophyll) and the chlorophyll recovery numerically, and the chlorophyll extract prepared under the conditions of the embodiment of the present invention is more purified than the chlorophyll extract prepared by the method of Comparative Example. Was found to be significantly higher. In addition, the chlorophyll recovery rate was not much different from the chlorophyll extract extracted using only the ethanol of Comparative Example 5, it was confirmed that the extract of the chlorophyll can be produced by the method of the present invention with good chlorophyll recovery.

Condition Purity (%, Chlorophyll Purity) Chlorophyll recovery (%) Example 1 90.14 9.13 Example 2 88.45 9.24 Example 3 87.44 9.21 Example 4 87.45 9.01 Comparative Example 1 69.43 9.56 Comparative Example 2 53.28 8.74 Comparative Example 3 46.39 9.54 Comparative Example 4 65.34 8.45 Comparative Example 5 54.23 8.34

< Experimental Example  2. Of Chlorophyll Extract Browning  Compare>

0.1 g of the chlorophyll dry matter prepared in Examples and Comparative Examples was dissolved in 100 g of soybean oil, 1 g of which was added to 99 g of soybean oil, stirred well, stored at room temperature for 30 days, and then a negative a value indicating greenness was measured using a colorimeter. The browning was observed by measuring the results. The results are shown in Table 2, and the lower the value, the better the result.

Condition Save 0 Day Chromaticity a Value Chromaticity a value after 30 days of storage Example 1 -15.35 -15.08 Example 2 -12.85 -11.28 Example 3 -10.81  -9.40 Example 4 -13.55 -13.02 Comparative Example 1 -6.81  -1.40 Comparative Example 2 -5.32  -2.23 Comparative Example 3 -2.84   5.62 Comparative Example 4 -4.42   2.34 Comparative Example 5 -4.34   1.32

As shown in Table 2, the chlorophyll extract of Example 1 extracted with 98% ethanol aqueous solution was found to have stronger greenness than the chlorophyll extracts of Examples 2 and 3 extracted with 80% and 70% ethanol, but after 30 days of storage The degree of browning was little affected by the alcohol concentration extracted.

In Comparative Example 1, which was not treated with copper sulfate, and Comparative Example 2, which was not treated with acid and base, the initial greenness values were also low, and it can be seen that browning proceeded significantly after 30 days of storage. In addition, the comparative example 5, which was dried without other pretreatment after extraction with 98% ethanol, had very low greenness (positive value), and browning occurred severely. In the method of Example, only solvent 30% ethanol and 50% In the case of Comparative Examples 4 and 5 in which chlorophyll extract was prepared with ethanol, the initial greenness was very low, and the degree of browning after 30 days was also severe.

<Example 1. Food preparation example>

1-1. Preparation of Cooking Seasonings

Chlorophyll extract of Example 1 according to the present invention to prepare a cooking seasoning to 0.02 ~ 1% by weight, respectively.

1-2. Manufacture of flour food products

The chlorophyll extract of Example 1 according to the present invention was added to the flour at 0.01 to 0.5% by weight, respectively, and bread, cake, cookies, crackers and noodles were prepared using this mixture.

1-3. Preparation of soups and gravy

The chlorophyll extract of Example 1 according to the present invention was added to the soup and broth to 0.01 to 0.1% by weight, respectively, to prepare meat products for health promotion, soup and noodles for noodles.

1-4. Manufacture of dairy products

The chlorophyll extract of Example 1 according to the present invention was added to milk at 0.01 to 0.1% by weight, respectively, and various dairy products such as butter and ice cream were prepared using the milk.

1-5. Preparation of Cooking Oil

1 mg of chlorophyll extract of Example 1 according to the present invention was put in 200 ml of soybean oil to prepare chlorophyll-containing cooking oil.

Claims (6)

(Step 1) grinding the dried tea leaves;
(Step 2) mixing the tea leaves and alcohol and leaching for 9-14 hours and then filtering;
(Step 3) adjusting the filtrate of step 2 with a basic solution to pH 10.0 ~ 12.0, and saponification reaction at 50 ~ 70 ℃ for 0.5 ~ 2 hours;
(Step 4) Take only the upper alcohol solution of the saponification reaction solution, mix the upper alcohol solution with petroleum ether in a volume ratio of 1-2: 1-2, remove the petroleum ether layer to remove pigments other than chlorophyll. step;
(Step 5) removing the petroleum ether layer from the mixture of step 4 and taking only the remaining alcohol solution, adjusting the pH to 3.0 to 5.0 with an acidic solution;
(Step 6) 5 to 10% by weight of an aqueous copper sulfate solution or zinc acetate solution to the alcohol solution adjusted to the pH of the step 5 to 3.0 to 5.0 to 10 to 60 minutes at 50-70 ℃ step; And
(7 step) drying the substituent under reduced pressure to prepare a dried product, filtering the precipitated inorganic salt by dissolving the dried product in alcohol, and then drying the filtered alcohol solution under reduced pressure again;
Method for producing a high quality chlorophyll extract comprising a. The method of claim 1,
Alcohol of the second step is a method for producing a high quality chlorophyll extract, characterized in that 80 to 100% ethanol aqueous solution or 80 to 100% aqueous methanol solution. The method of claim 1,
The basic solution in step 3 is a method for producing a high quality chlorophyll extract, characterized in that the aqueous sodium hydroxide solution or potassium hydroxide solution. The method of claim 1,
The acid solution in step 5 is a method for producing a high quality chlorophyll extract, characterized in that the concentrated hydrochloric acid or concentrated sulfuric acid. The method of claim 1,
The copper sulfate aqueous solution or zinc acetate aqueous solution of step 6 is a method for producing a high quality chlorophyll extract, characterized in that the 3-7% copper sulfate aqueous solution or 3-7% zinc acetate aqueous solution. Food containing a high quality chlorophyll extract prepared by the method of any one of claims 1 to 5.

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