KR102738487B1 - Method of manufacturing pomegranate seed extract for preventing hair loss and mist composition for preventing hair loss containing the extract - Google Patents

Abstract

The present invention relates to a method for producing a pomegranate seed extract for preventing hair loss and a mist composition for preventing hair loss containing the extract, and more specifically, to a method for producing a pomegranate seed extract for preventing hair loss and a mist composition for preventing hair loss containing the extract, which improves the condition of hair and prevents damage to the scalp caused by exposure to chemical products by producing an extract from pomegranate seeds and using a mist composition containing the pomegranate seed extract, thereby being effective in preventing hair loss.
The above invention is characterized by comprising a drying step of drying pomegranate seeds at a temperature of 40°C for 15 hours, a grinding step of grinding the pomegranate seeds dried through the drying step at a temperature of 100°C for 30 minutes to form pomegranate seed powder, a dissolution extraction step of mixing the pomegranate seed powder ground through the grinding step and 70% ethanol in a ratio of 1:3 in an aqueous solution, maintaining the temperature at 80°C, and extracting the filtrate by repeating extraction and filtration three times for 3 hours while reflux-cooling the 80°C aqueous solution, and a concentration step of obtaining a pomegranate concentrate by concentrating the filtrate extracted in the dissolution extraction step at a reduced pressure of 50°C.

Description

Method of manufacturing pomegranate seed extract for preventing hair loss and mist composition for preventing hair loss containing the extract

The present invention relates to a method for producing a pomegranate seed extract for preventing hair loss and a mist composition for preventing hair loss containing the extract, and more specifically, to a method for producing a pomegranate seed extract for preventing hair loss and a mist composition for preventing hair loss containing the extract, which improves the condition of hair and prevents damage to the scalp caused by exposure to chemical products by producing an extract from pomegranate seeds and using a mist composition containing the pomegranate seed extract, thereby being effective in preventing hair loss.

The pomegranate tree is a small fruit tree with deciduous foliage belonging to the Punicacea family, originally from Asia, especially the Middle East. It can grow to about 5 m, but most do not grow more than 1.5 m. The flowers are red-orange or orange, and vary from one to two. The fruit is edible and has been consumed since ancient times.

Synonyms include Punica spinosa, Punica florida, and Granatum puniceum.

Its fruit is the pomegranate, which is red-orange in colour and has a hard, tough rind (or peel) containing six to twelve membranous lobes containing numerous triangular seeds with transparent, juicy arils.

Pomegranates are rich in vitamins (B, C and D) and polyphenols in the form of tannins.

The bark extract exhibits astringent, antibacterial and anti-haemorrhagic properties.

Looking at previous patents on pomegranate extracts that extract and use components of these pomegranates, European Patent No. EP 138 419 discloses a topical treatment using pomegranate extracts to maintain the clarity of dyed hair for a long time.

International patent application WO 2007/004 229 discloses the use of the oily fraction of pomegranate seeds to promote skin regeneration.

French patent FR 2 730 408 relates to a composition, which may contain extracts of various fruits, including pomegranate, for preventing pigmentation of the skin by topical treatment.

Meanwhile, European Patent EP 1 523 895 discloses an oral composition comprising a combination of pomegranate extract and fatty acids, particularly conjugated linoleic acid (CLA) and OPC.

Also, US patent US 6 630 163 relates to a composition for treating skin diseases comprising fruit extracts, particularly pomegranate extracts, and US 2006/0 280 819 relates to a food supplement made from pomegranate seeds.

However, the conventional techniques of various pomegranate extracts as described above did not have a composition or a method for producing the same for improving the condition of hair, preventing damage to the scalp, and preventing hair loss.

The present invention relates to a method for producing a pomegranate seed extract for preventing hair loss and a mist composition for preventing hair loss using the extract, and more specifically, to a method for producing a pomegranate seed extract for preventing hair loss and a mist composition for preventing hair loss using the extract, which improves the condition of hair and prevents damage to the scalp caused by exposure to chemical products by producing an extract from pomegranate seeds and using a mist composition containing the pomegranate seed extract, thereby being effective in preventing hair loss.

Accordingly, the present invention has been made in consideration of the above-mentioned problems of the prior art, and the purpose of the present invention is to provide a method for producing a pomegranate seed extract for preventing hair loss, which is effective in preventing hair loss by improving the condition of the hair and preventing damage to the scalp caused by exposure to chemical products, and a mist composition for preventing hair loss using the extract.

In order to achieve the above-mentioned purpose, the present invention provides a method for producing a pomegranate seed extract for preventing hair loss, comprising a drying step of drying pomegranate seeds at a temperature of 40°C for 15 hours, a grinding step of grinding the pomegranate seeds dried through the drying step at a temperature of 100°C for 30 minutes to form pomegranate seed powder, a dissolution extraction step of mixing the pomegranate seed powder ground through the grinding step and 70% ethanol in a ratio of 1:3, maintaining the temperature of an aqueous solution at 80°C, and extracting a filtrate by repeating extraction and filtration three times for 3 hours while reflux-cooling the 80°C aqueous solution, and a concentration step of obtaining a pomegranate concentrate by concentrating the filtrate extracted in the dissolution extraction step at 50°C under reduced pressure.

The present invention provides a mist composition for preventing hair loss, which contains a pomegranate seed extract, comprising 89 to 91 wt% of distilled water free of organic and inorganic substances, 2 to 4 wt% of ethanol, a disinfectant that removes bacteria and mold, 2 to 4 wt% of triethanolamine that neutralizes fatty acids, adjusts and buffers pH, and dissolves oil, 0.5 to 2 wt% of polysorbate 20 that uniformly disperses liquids or solids that do not mix well in a liquid, 0.5 to 2 wt% of polysorbate 80 that uniformly disperses liquids or solids that do not mix well in a liquid and acts as an emulsifier and stabilizer, 0.5 to 2 wt% of glycerol that acts as a moisturizer and prevents deterioration of other substances, 0.5 to 1 wt% of poloximer that increases the water solubility of hydrophobic and oily substances or increases the miscibility of two substances with different hydrophobic properties, and 1 to 2 wt% of pomegranate seed extract that has estrogen and anti-estrogen effects.

As described above, the present invention has an effect of preventing hair loss by improving the condition of hair and preventing damage to the scalp caused by exposure to chemical products.

Although the present invention has been described and illustrated with specific preferred embodiments above, the present invention is not limited to the above-described embodiments, and various changes and modifications may be made by a person skilled in the art without departing from the spirit of the present invention.

Figure 1 is a schematic diagram showing the process of a method for manufacturing a pomegranate seed extract for preventing hair loss according to the present invention.

In order to fully understand the advantages of the present invention and the operational properties thereof and the objects achieved by the practice of the present invention, reference should be made to the accompanying drawings showing preferred embodiments of the present invention and the contents described in the accompanying drawings.

Hereinafter, the present invention will be described in more detail with reference to specific embodiments of the present invention. The following embodiments are provided only to explain the present invention in more detail, and the technical scope of the present invention is not limited thereby.

(Example)

The attached drawing 1 is a schematic diagram showing the process of a method for manufacturing a pomegranate seed extract for preventing hair loss according to the present invention.

As shown in Fig. 1, the method for producing a pomegranate seed extract for preventing hair loss of the present invention includes drying pomegranate seeds in a forced circulation dryer while maintaining a temperature of 40°C for 15 hours in the drying step.

Forced circulation method is effective for uniform heating and rapid drying as heat is transferred by a fan inside the forced circulation dryer. Since the air is circulated quickly, the time it takes to reach the target temperature is short, and the temperature uniformity and precision are excellent, so pomegranate seeds can be dried at the temperature at which the antioxidant component is measured to be the highest.

In order to grind the pomegranate seeds that have gone through this drying step, they are dried at a temperature of 100℃ during the grinding step and ground to a size of 100 mesh or larger for 30 minutes.

In order to dissolve the components in the pomegranate seed powder pulverized through the above pulverization step, in the dissolution extraction step, an aqueous solution in which pomegranate seed powder is mixed with 70% ethanol in a ratio of 1:3 is maintained at a temperature of 80°C, and the 80°C aqueous solution is cooled by reflux while repeating extraction and filtration three times for 3 hours to extract the filtrate.

At this time, a reflux condenser is used, and the solvent contained in the ethanol and pomegranate seed powder is prevented from vaporizing and evaporating from the aqueous solution mixed with pomegranate seed powder. The process of condensing the vaporized substance in the condenser inside the container and changing it into a filtrate is repeated three times.

The filtrate extracted in this way is concentrated through a concentration step to produce pomegranate seed extract.

The above concentration step is performed by placing the extracted filtrate in a rotating disc and rotating it to increase the surface area of the filtrate and reducing the internal environment pressure to 50℃ to increase the evaporation rate, thereby concentrating and separating the filtrate.

Such pomegranate seed concentrate can have antioxidant properties due to anthocyanins and substances such as ellagic acid and phytoestrogens.

A hair loss prevention composition containing pomegranate seed extract, which is composed of 89-91 wt% of distilled water that does not contain organic or inorganic substances, 2-4 wt% of ethanol, a disinfectant that removes bacteria or mold, 2-4 wt% of triethanolamine that neutralizes fatty acids, adjusts and buffers pH, and dissolves oil, 0.5-2 wt% of polysorbate 20 that uniformly disperses liquids or solids that do not mix well in a liquid, 0.5-2 wt% of polysorbate 80 that uniformly disperses liquids or solids that do not mix well in a liquid and acts as an emulsifier and stabilizer, 0.5-2 wt% of glycerol that acts as a moisturizer and prevents deterioration of other substances, 0.5-1 wt% of poloximer that increases the solubility of hydrophobic and oily substances or increases the miscibility of two substances with different hydrophobic properties, and 1-2 wt% of pomegranate seed extract that has estrogen and anti-estrogen effects. A mist composition can be obtained.

The hair loss prevention mist composition containing the pomegranate seed extract as described above is composed of 90 wt% of distilled water that does not contain organic and inorganic substances, 3 wt% of ethanol, which is widely used because of its high solubility in various substances, excellent volatility, and relatively low toxicity, and which denatures proteins, decomposes lipids, and thus kills organic substances, thereby eliminating most bacteria and fungi and many viruses that cause diseases and contamination, 2.5 wt% of triethanolamine, which is a strong alkaline substance used as an emulsifier and surfactant and which neutralizes fatty acids, adjusts pH, and plays a role in buffering, and also dissolves components or oils that are not sufficiently dissolved in water, 1 wt% of polysorbate 20, which uniformly disperses liquids or solids that do not mix well in a liquid, 1 wt% of polysorbate 80, which uniformly disperses liquids or solids that do not mix well in a liquid and acts as an emulsifier and stabilizer, and glycerol, which acts as a moisturizer and prevents deterioration of other substances. It is composed of 1 wt% of a poloximer, 0.5 wt% of a poloximer that increases the solubility of hydrophobic and oily substances or increases the miscibility of two substances with different hydrophobic properties, and 1 wt% of a pomegranate seed extract that has estrogen and anti-estrogen effects.

The following is an explanation of the results of measuring changes in scalp and hair condition after treatment with hair loss prevention mist using the above pomegranate seed extract.

(Example 1)

Effects of anti-hair loss mist composition containing pomegranate seed extract on scalp health.

A hair loss prevention mist composition containing pomegranate seed extract was applied to middle-aged women in their 40s to 60s who had fewer hair diseases due to pomegranate extract and were frequently exposed to chemical products due to perms and dyeing. The scalp moisture, scalp oil, scalp keratin, scalp pH, scalp erythema, scalp microorganisms, and scalp surface color were observed.

A hair loss prevention mist composition containing pomegranate seed extract was applied twice a day after shampooing, 2g each time, in the morning and evening, for a total of 4g.

Research subjects and data collection

The subjects of this study were middle-aged women in their 40s to 60s who visited for counseling without receiving any special care, and 22 who agreed to the purpose of the study were selected (IRB review number 7001355-201603-HR-113).

The subjects were selected as those who were not currently being treated for any disease, had not taken any specific medications, had not been on a diet, smoked, or drank alcohol within the past 6 months, had no specific illnesses, and agreed not to receive hair care or take antioxidants, health supplements, or other drugs during the experimental period.

A total of 22 subjects were selected for the study, divided into two groups: a control group with 11 subjects per group and a pomegranate seed extract scalp tonic experimental group.

Group 2 consists of people without any disease who eat their daily diet as usual without taking any dietary supplements. People with specific constitutions or allergies may experience hypersensitivity reactions, so they were selected as people without allergies.

In addition, in order to ensure the reliability of the experiment and minimize errors, measurements with a deviation range exceeding 30% were excluded, and to protect privacy and confidentiality, all research subjects were personally contacted by the researcher to explain the experiment for a total of 12 weeks, and then written consent and application forms were received before the experiment was conducted.

1. Measurement method and tools

Scalp measurements were performed before and after applying the pomegranate seed extract scalp tonic for 12 weeks.

Because the measurement value may vary depending on factors such as scalp moisture level, degree of wrinkles, degree of hair loss, posture, and body temperature, the laboratory temperature was 24–25°C and the relative humidity was 40–60%. The scalp was measured at the same point 10 cm back along the center line from the center of the facial scalp hairline of the subject 3 hours after shampooing, under the same time zone, lighting, and equipment. In order to minimize measurement errors, the same person performed the measurements from beginning to end.

A skin moisture meter (Skin Visiometer) (SV-600, Courage-Khazaka Electronic Co., Germany) was used to measure scalp keratin, a skin measurement device (Sebumeter) (SM 810, Courage-Khazaka Electronic Co., Germany) was used to measure scalp moisture and oil, a skin pH meter (SkinCheck-1, Hanna Instruments, Italy) was used to measure scalp pH, a colorimeter (Mexameter) (MX 18, Courage-Khazaka Electronic Co., Germany) was used to measure scalp erythema pigmentation, a bacteria meter (Lumitester) (TM PD-20, KIKOMAN, Japan) and a LuciPac (LuciPac W, KIKOMAN, Japan) were used to measure microbial counts, and a colorimeter (CR-400, Minolta, Japan) was used to measure the surface color of each scalp sample in color space (CIE The colorimetric system was measured using the lightness index (L*), redness (a*), and yellowness (b*) values.

1) Moisture content of the scalp

The moisture content of the scalp was measured using a skin measuring device (Sebumeter) (SM 810, Courage-Khazaka Electronic Co., Germany) and a moisture meter (CM-805).

When measuring moisture, the laboratory temperature was 24-25℃, the relative humidity was 40-60%, and the measurement was made 3 hours after shampooing, 10 cm behind the center line of the facial scalp hair border, at the same time (1-6 PM) and under the same lighting.

In addition, to minimize errors, measurements were repeated three times per individual at the same time, lighting, and location, and then the average was calculated. To ensure the reliability of the measurements, the average values with a deviation range exceeding 30% in each group were excluded, and the average ± standard deviation was calculated using SPSS for Windows version 23.0.

Measurements were taken twice, before and after applying the pomegranate seed extract scalp tonic.

2) Amount of oil in the scalp

The amount of scalp oil in the subjects was measured 3 hours after shampooing using a skin measuring device (Sebumeter) (SM 810, Courage-Khazaka Electronic Co., Germany).

In addition, a sebum measurement cassette (sebumeter cassette) was used to gently press a probe with a transparent film attached to the measurement area onto the skin surface for 20 seconds to absorb sebum, and then the probe was pressed onto the measurement port to measure the degree of light transmission.

For the measurement points, each individual was measured three times at points 5 cm behind, 10 cm behind, and 15 cm behind the center point of the facial scalp hair border without overlapping, and then the average value was calculated.

In addition, to ensure the reliability of the measurement values, the mean ± standard deviation was calculated using SPSS for Windows version 23.0, excluding the mean values whose deviation range exceeded 30% in each group.

The experiment was conducted before and after applying the pomegranate seed extract scalp tonic, and the measurements were taken in an environment where the laboratory temperature was 24–25°C and the relative humidity was 40–60%.

3) Scalp keratin

The scalp keratin content was measured using a Skin Visiometer (SV-600, Courage-Khazaka Electronic Co., Germany).

The measurement method involved placing a corneofix tape on the skin surface at a point 10 cm behind the center line of the scalp hair border, pressing it for 3 seconds, placing the removed dead skin cells in a cartridge, irradiating the cartridge with a light source, and photographing the image. The removed tape was then placed on the probe area, and the amount of removed dead skin cells was measured.

And the scalp keratin amount is expressed as a %, and the lower the % value, the lower the amount of keratin. The keratin amount was measured 3 times on the adjacent areas of the scalp for each person, and the mean ± standard deviation was calculated using SPSS for Windows version 23.0. An image with a horizontal and vertical area of 1㎠ where keratin was present was attached.

The experiment was conducted before and after applying the pomegranate seed extract scalp tonic, and the measurements were taken in an environment where the laboratory temperature was 24–25°C and the relative humidity was 40–60%.

4) pH of the scalp

The pH of the scalp was measured using a skin pH meter (SkinCheck-1, Hanna Instruments, Italy).

In addition, self-calibration was performed using a separate buffer solution (pH 4.0 / pH 7.0), and the measurement method was to measure the pH by placing the cylindrical ultrasonic probe of the scalp skin pH meter filled with buffer solution on the skin at a point 10 cm behind the center point of the facial scalp hair border of the experimental subject 3 hours after shampooing.

In order to minimize the error, the measurements were repeated three times per individual at the same time, lighting, and location, and then the average was calculated. In order to ensure the reliability of the measurements, if the deviation range of the average value exceeded 30% in each group, the average value ± standard deviation (Mean ±) was calculated using SPSS version 23.0 for Windows. The measurements were conducted before and after applying the pomegranate seed extract scalp tonic, and the measurements were taken in an environment with a laboratory temperature of 24–25°C and a relative humidity of 40–60%.

5) Amount of erythema pigment on the scalp

The amount of erythema pigment in the scalp was measured using a colorimeter (Mexameter) (MX 18, Courage-Khazaka Electronic Co., Germany).

For MX 18, it is a narrow-band reflectance spectrophotometer that emits light such as 568 nm (green), 660 nm (red), and 880 nm (infrared) from a probe and measures the light reflected from the skin, which is determined by the content and distribution of hemoglobin and melanin.

For the measurement method, the scalp was touched with a colorimeter (Mexameter MX 18) at a point 10 cm behind the midline from the center point of the facial scalp hair border, and the erythema index (EI) of the scalp was measured through the sensor within 1 second.

In addition, to minimize errors, measurements were repeated three times per individual at the same time, lighting, and location, and then the average was calculated. To ensure the reliability of the measurements, the average values with a deviation range exceeding 30% in each group were excluded, and the average ± standard deviation was calculated using SPSS for Windows version 23.0.

The experiment was conducted before and after applying the pomegranate seed extract scalp tonic, and the measurements were taken in an environment where the laboratory temperature was 24–25°C and the relative humidity was 40–60%.

6) Microbial load on the scalp

The amount of microorganisms on the scalp was measured using a bacteriometer (Lumitester) (TM PD-20, KIKOMAN, Japan) and LuciPac (LuciPac W, KIKOMAN, Japan).

The measurement principle is to measure ATP consumption by luciferase, and the presence of microorganisms can be determined based on luminescence.

The measurement method was to use a disposable LuciPac (Swab test) and rub the scalp 3 times from a point 10 cm behind the center of the hairline of the subject's face 3 hours after shampooing, then dilute the tip of the swab in the solution, shake it well 10 times, and measure using a bacteria meter (Lumitester).

In addition, to minimize errors, measurements were repeated three times per individual at the same time, lighting, and location, and then the average was calculated. To ensure the reliability of the measurements, the average values with a deviation range exceeding 30% in each group were excluded, and the average ± standard deviation was calculated using SPSS for Windows version 23.0.

The experiment was conducted twice, before and after applying the pomegranate seed extract scalp tonic, and the measurements were taken in an environment where the laboratory temperature was 24–25°C and the relative humidity was 40–60%.

7) Surface color of the scalp

The surface color of each scalp sample was measured using a colorimeter (CR-400, Minolta, Japan) in terms of the lightness index (L*), redness (a*), and yellowness (b*) of the CIE Lab color system. The measurement specifications were based on the General Principles for Measurement of Surface Color of Textile Dyeing Fastness in the Korean Industrial Standards (ISO 105-J01, 2003). The measurement point was 1 cm from the hair root of the scalp, and the measurements were repeated three times for each subject and the average value was calculated. In addition, in cases where the average value exceeded 30% in each group, the average value ± standard deviation was calculated using SPSS version 23.0 for Windows.

The experiment was conducted before and after applying the pomegranate seed extract scalp tonic, and the measurements were taken in an environment where the laboratory temperature was 24–25°C and the relative humidity was 40–60%.

2. Data Analysis

The collected data were statistically processed using SPSS version 23.0 for Windows (PASS for Windows version 23.0).

Continuous variables are expressed as mean ± standard deviation, and categorical variables are expressed as frequency and percentage (%).

If the significance probability p value is less than 0.05 ( p < 0.05), it is considered statistically significant.

The statistical method for data analysis in this example is as follows.

First, to verify whether the control group and experimental group were composed of homogeneous groups, an independent sample t-test was conducted on scalp moisture, oil, keratin, pH, erythema, microorganisms, and surface color.

Second, a paired t-test was performed to determine the differences in scalp moisture, oil, keratin, pH, erythema, microorganisms, and surface color between the control group and the experimental group before and after pomegranate seed extract treatment.

3. Differences in scalp test amount according to pomegranate seed extract

1) Scalp moisture content

The health of the scalp, which can be considered the basis of hair, is very important for hair growth, so external damage to the scalp or accumulation of waste matter can hinder hair growth and health, and cause hair loss. The human body is composed of approximately 70% water, and the keratin layer of the epidermis contains 10-20% water.

The stratum corneum contains intercellular lipids, including natural moisturizing factors composed of ceramides and amino acids, which play a role in preventing moisture loss in the stratum corneum (Lee Ae-young, 2015). The moisture content of the scalp stratum corneum is important for maintaining a healthy scalp.

Moisture keeps hair soft, and lack of moisture causes hair to become dry and dull, leading to hair damage.

When hair damage is severe, the hair's ability to retain moisture is weakened and the moisture content decreases, so moisture content becomes the standard for hair damage.

Normal hair generally has a moisture content of around 15%, oily hair around 20%, and dry hair less than 10%.

Abnormal symptoms of dry scalp, which is a lack of moisture in the scalp, include itchiness due to insufficient sebum secretion, and the acid film is damaged, causing the scalp to feel tight, resulting in itching, inflammation, skin peeling, and scalp damage.

The pomegranate seed extract mist composition was divided into an experimental group and a control group to compare the effects of pomegranate seed extract through changes in moisture before and after the experiment in scalp care using a scalp tonic, and the experimental group was treated with a pomegranate seed extract scalp tonic. After 12 weeks of clinical trials, the results are shown in Table 1.

The scalp moisture of the control group decreased from 11.01±2.27 before the experiment to 3.63±1.72 after the experiment, which was not significant (p>.05). The experimental group decreased from 11.14±19.05 before the experiment to 10.86±10.76, which was not significant (p>.05). In addition, the scalp moisture of the control group (11.01±2.27) before the experiment and the scalp moisture of the experimental group (11.14±19.05) showed a difference between the two groups, but was not significant (p>.05), and the scalp moisture of the control group (3.63±1.72) and the scalp moisture of the experimental group (10.86±10.76) after the experiment treated with pomegranate seed extract showed a significant difference (p<.05). That is, the experimental group that applied the pomegranate seed extract scalp tonic showed a scalp moisture content decrease of approximately 67% less than the control group, indicating that the moisture was maintained.

Differences in scalp moisture before and after pomegranate seed extract in the control and experimental groups Scalp water supply Control group Experimental group t (p) Before the experiment 11.01±2.27 11.14±19.05 -0.937(.361) After the experiment 3.63±1.72 10.86±10.76 -2.097 ^* (.050) t (p) 2.024(.058) 0.041(.968)

The above results are due to the fact that the estrogen in pomegranate seeds increases collagen and hyaluronic acid in the skin in the human body, which has a positive effect on skin elasticity and moisture.

Therefore, it was confirmed that hair loss prevention mist containing pomegranate seed extract is a good way to increase the moisture content of the scalp for scalp care to prevent hair loss in menopausal women.

2) Scalp Oil Content

The oil of the scalp is measured as sebum excretion, a substance secreted through the sebaceous glands, and comes to the skin surface in the form of pure oil containing triglycerides (40-60%), wax esters (19-26%), and squalene (11-15%).

In alopecia areata, seborrheic dermatitis is the most common concomitant disease (Yoo Gwang-ho et al., 2009), as the sebaceous glands are activated due to increased androgens (Chen et al., 1996; Zhang et al., 2008). In the case of seborrheic dermatitis, the sebaceous glands of the scalp secrete excessive sebum, which alkalizes the sebum film of the scalp and increases the proliferation of microorganisms. The increased microorganisms decompose the sebum and then secrete irritating substances to the scalp.

In order to compare the effects of pomegranate seed extract through changes in sebum before and after scalp care using hair loss prevention mist containing pomegranate seed extract, the experimental group and the control group were divided and treated with pomegranate seed extract scalp tonic. After 12 weeks of clinical trials, the results are shown in Table 2.

The scalp oil content of the control group decreased from 209.80±96.32 before the experiment to 194.00±82.98 after the experiment, which was not significant (p>.05). The experimental group decreased from 210.70±57.14 before the experiment to 84.80±42.91, which was significant (p<.001). In addition, the scalp oil content of the control group (209.80±96.32) before the experiment and the experimental group (210.70±57.14) showed a difference between the two groups, but was not significant (p>.05), and the scalp oil content of the control group (194.00±82.98) and the experimental group (84.80±42.91) after the experiment treated with pomegranate seed extract showed a significant difference (p<.01). That is, the experimental group that applied the hair loss prevention mist containing pomegranate seed extract showed a decrease in scalp oil content of approximately 60%.

Differences in scalp oil before and after pomegranate seed extract in the control and experimental groups Scalp water supply Control group Experimental group t (p) Before the experiment 209.80±96.32 210.70±57.14 -1.268(.221) After the experiment 194.00±82.98 84.80±42.91 3.696(.002 ^** ) t (p) -0.701(.492) 5.571(.000 ^*** )

Regulating scalp oiliness, collagen intake by middle-aged women in their 40s to 60s increases dermal collagen synthesis, and sebum secretion is greatly reduced by restoring epidermal cell metabolism. A combination diet of vinegar and collagen is more effective than a collagen-only diet.

And it was revealed that the excessive sebum secreted by middle-aged women was reduced as the skin oil and moisture content was regulated by collagen intake, and that a combined diet of collagen and enzymes had a positive effect on the scalp, especially helping to reduce oil content. Most of the previous studies were related to the intake of dietary supplements.

In addition, it is thought that the scalp of middle-aged women in their 40s to 60s has an increased amount of sebum because estrogen deficiency and excessive androgen secretion promote the activity of the sebaceous glands due to a decline in ovarian function, usually around 4 to 5 years before and after menopause (Dobransky et al., 1987).

Therefore, it was estimated that the application of hair loss prevention mist containing pomegranate seed extract not only had a positive effect on scalp improvement, but also had a complex effect, reducing scalp oil. In other words, it was confirmed that the scalp tonic containing pomegranate seed extract was a good way to reduce scalp oil in scalp care for hair loss prevention.

3) Scalp keratin content

If the cycle of keratinization, in which keratin cells in the basal layer of the scalp move to the stratum corneum and fall off, becomes too fast, the cell regeneration ability and skin defense ability decrease, and if it becomes too slow, the shedding rate of keratin decreases, and if keratin remains in the stratum corneum for a long time, the stratum corneum thickens and hardens, blocking pores.

Representative lesions of dyskeratosis with an accelerated cycle include scalp psoriasis, and representative lesions of hyperkeratosis with a slow cycle include hyperpigmentation and acne.

Hyperkeratosis and dyskeratosis are lesions that cause secondary hair loss. In addition, hair loss may occur when the pore opening is blocked by excessive sebum secretion from the sebaceous glands, preventing normal skin shedding.

In order to compare the effects of pomegranate seed extract through changes in keratin before and after scalp care using hair loss prevention mist containing pomegranate seed extract, the experimental group and the control group were divided and treated with pomegranate seed extract scalp tonic. After 12 weeks of clinical trials, the results are shown in Table 3.

The scalp keratin content of the control group decreased from 12.92±3.05 before the experiment to 10.92±5.79 after the experiment, which was significant (p<.05). The experimental group decreased from 13.00±10.10 before the experiment to 3.84±2.93, which was significant (p<.05). In addition, the scalp keratin content of the control group (12.92±3.05) and the experimental group (13.00±10.10) before the experiment showed no significant difference between the two groups (p>.05), and the scalp keratin content of the control group (10.92±5.79) and the experimental group (3.84±2.93) after the experiment treated with pomegranate seed extract showed a significant difference (p<.01). In other words, it was found that the experimental group that was applied with the pomegranate seed extract showed a decrease in scalp keratin content of approximately 71%.

Differences in scalp keratin before and after pomegranate seed extract in the control and experimental groups Scalp water supply Control group Experimental group t (p) Before the experiment 12.92±3.05 13.00±10.10 -2.182(.053) After the experiment 194.00±82.98 84.80±42.91 3.453(.003 ^** ) t (p) -0.701(.492) 5.571(.000 ^*** )

The increase in scalp keratin in the control group is thought to be due to the fact that in the case of middle-aged women, excessive secretion of androgens promotes the activity of the sebaceous glands, which causes the scalp to become alkaline due to excessive sebum secretion, causing the keratin on the scalp to swell and become inflated, and then normal skin shedding does not occur, resulting in the accumulation and increase of keratin.

When middle-aged women in their 40s to 60s begin menopause, the synthesis of elastin and collagen, which are related to skin tension and elasticity, gradually decreases, and the amount of dead skin cells increases due to complex causes such as dry scalp caused by lack of moisture, stress, and impaired blood circulation.

Therefore, it was confirmed that hair loss prevention mist containing pomegranate seed extract is a good way to reduce the amount of dead skin cells on the scalp for scalp care to prevent hair loss.

4) Scalp pH content

The pH of the skin is the pH of the protective film on the skin surface that is formed by a mixture of sweat and sebum secreted to the skin surface. This acidic film suppresses excessive growth of microorganisms on the skin surface and protects the skin from infection, irritation, and itchiness.

The pH of a normal scalp is slightly acidic, around 4.5-5.5, and when the pH of the scalp becomes alkaline, it promotes the growth of basophilic scalp flora.

In order to compare the effects of pomegranate seed extract through changes in pH before and after the experiment in scalp care using hair loss prevention mist containing pomegranate seed extract, the experimental group was divided into an experimental group and a control group, and the experimental group was treated with pomegranate seed extract scalp tonic. After 12 weeks of clinical trials, the results are shown in Table 4.

The scalp pH of the control group decreased from 5.94±0.26 before the experiment to 5.80±0.41 after the experiment, which was not significant (p>.05). The experimental group decreased from 5.99±0.39 before the experiment to 5.33±0.32, which was significant (p<.001). In addition, the scalp pH of the control group (5.94±0.26) before the experiment and the scalp pH of the experimental group (5.80±0.41) showed a difference between the two groups, but was not significant (p>.05), and the scalp pH of the control group (5.80±0.41) and the experimental group (5.33±0.32) after the experiment treated with pomegranate seed extract showed a significant difference (p<.01). In other words, the scalp pH of the control group was unchanged, but the scalp pH of the experimental group decreased by about 11%.

Differences in scalp pH before and after pomegranate seed extraction in the control and experimental groups Scalp water supply Control group Experimental group t (p) Before the experiment 5.94±0.26 5.99±0.39 -1.196(.247) After the experiment 5.80±0.41 5.33±0.32 2.888 ^** (.010) t (p) 0.071(.944) 4.187(.001 ^*** )

Looking at papers related to skin pH control, the effect of seaweed diet on acne-prone skin was confirmed, and the pH in the forehead, cheeks, nose, and under the eyes, excluding the chin, increased from 5.80-5.98 before the test to 5.12-5.36 after the test, approaching the slightly acidic state of normal skin, whereas there was no significant change in the control group.

The pH changes in the neck were observed by applying a Cheonggukjang diet and back massage to middle-aged women. The pH changes in the facial skin were observed after applying a crushed fermented Cheonggukjang diet and back meridian massage. All of them showed a pH reduction control effect.

As such, most studies on scalp pH are based on the effects of dietary supplements.

In the case of the experimental group, it can be predicted that the phytoestronens component of pomegranate seeds has antioxidant, anti-inflammatory, and anticancer effects (Lansky et al., 2000; Verardo et al., 2014), which increases the immunity of the scalp and normalizes the antibacterial defense function, thereby affecting the pH concentration. This pH-reducing effect can be seen to have improved the scalp by restoring the acid film function of the skin.

Therefore, it was confirmed that hair loss prevention mist containing pomegranate seed extract is a good method for scalp care to prevent hair loss by reducing the pH of the scalp and helping to keep it within the normal range.

5) Scalp Microbiota

Among the microorganisms detected on the surface of the forehead, armpits, chest, and scalp of normal Koreans, the most common was Propionibacterium acnes, accounting for 84% of the total, followed by aerobic diphtheroids at 14%, and coagulases-type Staphylococci and Micrococci at 2%.

Sebum secreted from the sebaceous glands of the hair follicles increases under the action of androgen hormones, and the greater the amount of sebum on the scalp, the greater the number of microorganisms present.

Microorganisms attached to the scalp decompose organic compounds such as waste products or pollutants on the scalp, generating volatile odorous substances and influencing various scalp diseases.

In order to compare the effects of pomegranate seed extract through changes in microbial counts before and after scalp care using hair loss prevention mist containing pomegranate seed extract, the experimental group was divided into an experimental group and a control group, and the experimental group was treated with pomegranate seed extract mist. After 12 weeks of clinical trials, the results are shown in Table 5.

The scalp microbiota of the control group increased from 1312.40±914.827 before the experiment to 1454.70±855.27 after the experiment, which was not significant (p>.05). The experimental group decreased from 1299.60±460.57 before the experiment to 413.20±378.09, which was significant (p<.01). And the scalp microbiota of the control group before the experiment (1312.40±914.82) and the scalp microbiota of the experimental group (1299.60±460.57) showed a difference between the two groups, but it was not significant (p>.05), and the scalp microbiota of the control group after the experiment treated with pomegranate seed extract (1454.70±855.27) and the scalp microbiota of the experimental group (413.20±378.09) showed a significant difference (p<.01).

That is, the experimental group that applied hair loss prevention mist containing pomegranate seed extract showed a decrease in scalp microorganism count of approximately 62%.

Differences in scalp microflora before and after pomegranate seed extract in the control and experimental groups Scalp water supply Control group Experimental group t (p) Before the experiment 1312.40±914.82 1299.60±460.57 0.685(.502) After the experiment 1454.70±855.27 413.20±378.09 3.522(.002 ^** ) t (p) -0.359(.724) 3.595(.002 ^** )

The increase in the scalp microbiota in the control group was thought to be due to the increased sebum secretion caused by excessive androgen hormone secretion in middle-aged women in their 40s to 60s, which also promoted the microbiota by creating favorable conditions for microbial growth.

This indicates that when the sebaceous glands of the scalp are activated and the amount of oil increases excessively, the sebum film becomes alkaline, which increases the proliferation of microorganisms.

When looking at studies related to improving skin microbiota, it was confirmed that an aqueous solution containing phytosphingosine was applied to acne-prone skin and that it was effective in treating inflammatory acne. It was said that it had excellent antibacterial properties against skin microbes and thus had sufficient value as an antibacterial ingredient.

The skin cosmetic efficacy was elucidated through the antioxidant effect of peat, and it was reported that the antioxidant effect was the highest in Finnish Sphagnum peat.

In the experimental group that applied the hair loss prevention mist containing pomegranate seed extract, the amount of microorganisms on the scalp significantly decreased. It is thought that the pomegranate seed extract improved the condition of the scalp by regulating the oil and moisture, keratin, pH, and erythema pigment of the scalp of middle-aged women in their 40s to 60s, and also reduced and controlled the amount of microorganisms.

In this way, it can be seen that a healthy scalp maintains a slightly acidic state, maintains homeostasis of the permeability barrier function, and also plays an important role in the antibacterial barrier function, inhibiting the growth of pathogenic microorganisms.

Therefore, it was confirmed that hair loss prevention mist containing pomegranate seed extract is a good way to reduce the amount of microorganisms on the scalp for scalp care to prevent hair loss.

6) Scalp surface color

The internal structure of hair is made up of three layers, similar to the cross-section of a pencil. Each layer is made up of keratin protein, but each has different properties.

The double cortex is an important part that determines the properties of hair and contains granular melanin pigments that determine the color of the hair, so the color of the hair is determined by these melanin pigments.

The melanin granules that determine the color of hair are eumelanin and pheomelanin. Both types of hair pigments exist together in hair, and the color of hair in its natural state is determined by the ratio of eumelanin and pheomelanin.

To determine the effect of pomegranate seed extract on the surface color of the scalp of menopausal women, the surface color of the hair samples was measured using a colorimeter in terms of the lightness index (L*), redness (a*), and yellowness (b*) of the CIE Lab colorimetric system before and after the test (Table 9-11). In the colorimeter, lightness (L*) refers to brightness and is expressed as a number from 0 to 100, with 0 representing black and 100 representing white. In addition, redness (a*) is on the axis from green to red, with a +a value indicating redness and a -a value indicating greenness. In addition, yellowness (b*) is on the axis from yellow to blue, with a +b value indicating yellowness and a b value indicating blueness.

First, the redness of the scalp surface color of the control group decreased from 4.29±1.27 before the experiment to 3.20±1.15 after the experiment, which was not significant (p>.05). The experimental group decreased from 4.30±2.10 before the experiment to 2.24±1.25, which was also significant (p<.05). In addition, the redness of the scalp surface color of the control group (4.29±1.27) before the experiment and the redness of the scalp surface color of the experimental group (4.30±2.10) showed a difference between the two groups, but was not significant (p>.05), and the redness of the scalp surface color of the control group (3.20±1.15) after the experiment treated with pomegranate seed extract showed a difference between the two groups, but was not significant (p>.05). That is, in the experimental group, the scalp surface color redness to which the pomegranate seed extract scalp tonic was applied was reduced by approximately 48%, indicating that the amount of scalp erythema was improved. Unlike the redness, no significant results were measured for the brightness and yellowness of the scalp surface.

Next, the yellowness of the scalp surface color of the control group decreased from 13.85±4.01 before the experiment to 13.41±4.15 after the experiment, which was not significant (p>.05). The experimental group increased from 15.40±3.61 before the experiment to 16.29±3.57, which was not significant (p>.05). In addition, the yellowness of the scalp surface color of the control group before the experiment (13.85±4.01) and the experimental group (15.40±.3.61) showed a difference between the two groups, but was not significant (p>.05), and the yellowness of the scalp surface color of the control group (13.41±4.15) and the experimental group (16.29±3.57) after the experiment treated with pomegranate seed extract showed a difference between the two groups, but was not significant (p>.05). That is, it was found that the pomegranate seed extract scalp tonic did not have a significant effect on hair loss prevention through improvement of the yellowness of the scalp surface color.

In addition, the brightness of the scalp surface color of the control group increased from 38.20±7.57 before the experiment to 36.41±7.56 after the experiment, which was not significant (p>.05). The experimental group increased from 38.07±5.58 before the experiment to 40.02±4.58, which was not significant (p>.05). In addition, the brightness of the scalp surface color of the control group before the experiment (38.20±7.57) and the brightness of the scalp surface color of the experimental group (38.07±.5.58) showed a difference between the two groups, but was not significant (p>.05), and the brightness of the scalp surface color of the control group (36.41±7.56) and the experimental group (40.02±4.58) after the experiment treated with pomegranate seed extract showed a difference between the two groups, but was not significant (p>.05). That is, it was found that the pomegranate seed extract scalp tonic did not have a significant effect on hair loss prevention through improving the brightness of the scalp surface color.

By applying it, there was no significant difference in the surface color of the scalp, which is the lightness index (L*) and yellowness index (b*), but there was a significant difference in the redness index (a*) (p<.05). It can be said that pomegranate seed extract reduces the redness index of the scalp surface color, which improves scalp problems such as blood circulation and blood vessels.

7) Scalp erythema

The scalp is the organ most susceptible to the effects of ultraviolet rays due to the destruction of the ozone layer, and erythema is a phenomenon that occurs in the epidermal and dermal cells when exposed to ultraviolet rays when the blood vessels of the skin dilate or when there is inflammation in the skin, and refers to the skin becoming congested and red due to the dilation of the small blood vessels in the dermal papilla and the vascular plexus of the papilla.

In other words, skin erythema refers to a state in which the ability of capillary walls to return to their original state deteriorates due to expansion of blood vessels caused by internal and external stimuli when chronically exposed to climate conditions with extreme temperature differences or irritating environments.

This type of sensitive scalp easily shows inflammatory signs such as congestion and erythema, and when the inflammation is severe, it can lead to hair loss.

In this regard, as a result of examining the management effect of sensitive scalp, it was estimated that there are various factors such as individual skin management method and degree of sunlight exposure as the cause of the increase in skin erythema index in the changes in oil and moisture, erythema, and melanin index of skin to which beta-glucan was applied.

In order to compare the effect of pomegranate seed extract through changes in scalp erythema index before and after scalp care using hair loss prevention mist containing pomegranate seed extract, the experimental group was divided into an experimental group and a control group, and hair loss prevention mist containing pomegranate seed extract was applied to the experimental group. After 12 weeks of clinical trials, the results are shown in Table 6.

The scalp erythema of the control group increased from 102.70±33.81 before the experiment to 107.10±29.55 after the experiment, which was not significant (p>.05). The experimental group decreased from 104.00±33.02 before the experiment to 76.40±26.77, which was not significant (p>.05). In addition, the scalp erythema of the control group (102.70±33.81) before the experiment and the scalp erythema of the experimental group (104.00±33.02) showed a difference between the two groups, but was not significant (p>.05), and the scalp erythema of the control group (107.10±29.55) and the experimental group (76.40±26.77) after the experiment treated with pomegranate seed extract showed a significant difference (p<.05). That is, the experimental group that applied the hair loss prevention mist containing pomegranate seed extract showed a decrease in the erythema index by approximately 27%.

Differences in scalp erythema before and after pomegranate seed extract treatment in the control and experimental groups Scalp water supply Control group Experimental group t (p) Before the experiment 102.70±33.81 104.00±33.02 -1.091(.290) After the experiment 107.10±29.55 76.40±26.77 2.435 ^* (.026) t (p) -1.366(.189) 2.053(.055)

In the case of the control group of this experiment, it was possible to infer that as the weather got warmer, the capillary walls of the scalp expanded and sebum secretion increased, which led to an increase in erythema pigmentation due to pigmentation caused by exposure to ultraviolet rays. This is because skin with sensitivity due to erythema frequently has forms such as capillary dilated skin and erythema skin with repeated reddening, and seborrheic scalp, dandruff scalp, hair loss scalp, and inflammatory scalp have a lot of sebum and dead skin cells, and most of them are accompanied by erythema and inflammation.

In addition, erythema refers to a skin condition in which the amount of blood in the skin blood vessels increases and the skin temporarily turns red, and skin reactions due to ultraviolet ray exposure typically include erythema and pigmentation, which may occur for several hours and then completely disappear, or inflammation accompanied by pain may occur.

Therefore, it was confirmed that hair loss prevention mist containing pomegranate seed extract is a good method for reducing scalp erythema in scalp care to prevent hair loss.

A hair loss prevention mist containing pomegranate seed extract was manufactured and applied to test subjects. Differences in scalp moisture, oil, keratin, pH, erythema, microorganisms, and surface color were examined.

A total of 22 subjects were selected for the study, with 11 subjects in each group divided into two groups: a control group and a pomegranate seed extract scalp tonic experimental group. The experimental group scalp tonic was manufactured to contain 1.0% pomegranate seed extract, and the control group scalp tonic was applied with the same amount of purified water as the pomegranate seed extract used in the experimental group scalp tonic. The experiment was conducted.

First, there was no difference in scalp moisture between the control group and the experimental group before the experiment, and there was a significant difference between the control group and the experimental group after applying the pomegranate seed extract scalp tonic (p<.05). This means that the experimental group that applied the pomegranate seed extract scalp tonic maintained scalp moisture by reducing scalp moisture by about 67% less than the control group.

Second, there was no difference in the amount of scalp oil between the control group and the experimental group before the experiment, but there was a significant difference between the control group and the experimental group after applying the pomegranate seed extract scalp tonic (p<.01). This means that the experimental group that applied the pomegranate seed extract scalp tonic showed an improvement in scalp oil, with a decrease of approximately 60% in scalp oil compared to the control group.

Third, there was no difference in the scalp keratin content between the control group and the experimental group before the experiment, but there was a significant difference between the control group and the experimental group after applying the pomegranate seed extract scalp tonic (p<.01). This means that the experimental group that applied the pomegranate seed extract scalp tonic had approximately 71% decrease in scalp keratin content.

Fourth, there was no difference in scalp pH between the control group and the experimental group before the experiment, but there was a significant difference between the control group and the experimental group after applying the pomegranate seed extract scalp tonic (p<.01). This means that there was no change in the control group, but the experimental group's scalp pH improved by about 11%, which allowed the scalp pH to fall within the normal volume range.

Fifth, there was no difference in the scalp microbiota between the control group and the experimental group before the experiment, but there was a significant difference between the control group and the experimental group after applying the pomegranate seed extract scalp tonic (p<.01). This means that the experimental group that applied the pomegranate seed extract scalp tonic showed a decrease in the scalp microbiota by approximately 62%.

Sixth, there was no difference in the lightness index (L*) and yellowness (b*) of the scalp surface color between the control group and the experimental group before the experiment, but there was a significant difference in the redness (a*) (p<.05). It was found that the redness (a*) of the surface color after applying the pomegranate seed extract scalp tonic in the experimental group was reduced by approximately 48%, indicating that the erythema index was improved.

Seventh, there was no difference in scalp erythema between the control group and the experimental group before the experiment, but there was a significant difference between the control group and the experimental group after applying the pomegranate seed extract scalp tonic (p<.05). This means that the experimental group that applied the pomegranate seed extract scalp tonic showed a decrease in the erythema index by approximately 27%.

Example 2)

Effects of pomegranate seed extract on hair health

Apply 2g of anti-hair loss mist containing pomegranate seed extract twice a day, every morning and evening, after shampooing, for a total of 4g.

Hair samples were collected before and after applying the pomegranate seed extract scalp tonic for 12 weeks.

From the center of the scalp-hair border of the experimental subject's face, about 200 hairs were collected from a range of about 0.5 cm from the scalp as close to the hair root as possible, around 20 cm behind the center line. The length was about 15 cm. Since the measurement value may vary depending on the level of oil and moisture in the scalp and hair and the shampoo time, the hair of the subject was collected 3 hours after shampooing at the same time (1-5 PM). In addition, before the measurement, all sample hair was washed with a neutral shampoo with pH 7 using purified water to remove foreign substances and dust, and then naturally dried at room temperature of about 25℃.

1. Research subjects and data collection

The subjects of this study were selected from 22 middle-aged women who visited for consultation without receiving special care and agreed to the purpose of the study (IRB review number 7001355-201603-HR-113). Those who are currently not receiving treatment for any disease, have not taken any specific medication, dieted, smoked, or drank alcohol within the past 6 months, have no specific disease, and agree not to receive hair care during the experimental period and not to take health supplements including antioxidants or other drugs are selected as subjects.

A total of 22 subjects were selected for the study, with 11 subjects per group divided into two groups: the control group and the pomegranate seed extract scalp tonic experimental group. Group 2 consists of people without any disease who eat their daily diet as usual without taking any dietary supplements. Those with specific constitutions or allergies are selected as those without allergies as they may have hypersensitivity reactions. In addition, in order to minimize the reliability and error of the experiment, measurements with a deviation range of more than 30% after measurement are excluded. From August 2, 2016 to August 1, 2017, for a total of 12 weeks, the researcher personally explained the experiment to all subjects in person to protect their privacy and confidentiality, and the experiment was conducted after receiving written consent and application forms.

2. Measurement method and tools

Hair measurements were conducted separately for hair samples collected before applying hair loss prevention mist containing pomegranate seed extract and for hair samples collected after 12 weeks of application. The measurements were performed under the same time zone, lighting, and equipment in an environment with a laboratory temperature of 24–25°C and a relative humidity of 40–60%. To minimize measurement errors, the same person performed the measurements from beginning to end. In order to visually observe hair growth, the scalp was photographed at 200x magnification using a digital microscope (AM3013T, Dino-lite, Taiwan) and compared.

In addition, the number of hairs per unit area (3.6 mm × 2.8 mm) and the number of hairs per hair follicle were measured at 200x magnification using a contact scalp hair diagnostic device (Multi-cam 1000, Bandomedical, Korea), and the hair thickness was measured 10 times using a digital micrometer (MDC-25PJ, Mitutoyo Co., Japan).

1) Number of hairs

In order to visually observe hair growth, the scalp of the experimental subjects 3 hours after shampooing was photographed at 200x magnification using a digital microscope (AM3013T, Dino-lite, Taiwan) to compare before and after the experiment. For photography, first, the scalp hair of the subject's facial scalp hair border was cut cleanly in a circle with a diameter of 1 cm, 10 cm behind the center line, as close to the scalp as possible, as if shaving, and then the digital microscope was placed on the area and photographed. The same area was rephotographed after one week to check the condition of hair growth. After this, the subjects began taking the dietary supplement, and after 12 weeks, the same area of the scalp was cleanly cut as if shaving in the same manner, photographed, and then photographed again after one week to observe hair growth.

To measure the number of hairs per unit area of the scalp (3.6 mm × 2.8 mm), the number of hairs was measured using a contact scalp hair diagnostic device (Multi-cam 1000, Bandomedical, Korea) with a 200x magnification lens on the scalp 10 cm behind the center point of the facial scalp hair border of the subject 3 hours after shampooing.

To ensure the reliability of the measurement values, the measurements were repeated three times for each subject and the average was calculated. In addition, in the case of the average value with a deviation range exceeding 30% in each group, the mean ± standard deviation (Mean ± SD) was calculated using SPSS for Windows version 23.0. The measurements were performed twice, before and after applying the pomegranate seed extract scalp tonic, and the measurements were taken in an environment with a laboratory temperature of 24–25°C and a relative humidity of 40–60%.

2) Hair thickness

Hair thickness was measured by collecting 10 hair samples from each subject using a digital micrometer (MDC-25PJ, Mitutoyo Co., Japan).

The measurement point was 1 cm from the hair root, and in order to minimize errors and ensure the reliability of the experiment, measurements were repeated 10 times for each individual. After excluding the measurement values with a deviation range exceeding 30%, the average value was calculated.

In addition, in each group, the mean value ± standard deviation (Mean ± SD) was calculated using SPSS for Windows version 23.0, excluding the average value whose deviation range exceeded 30%. The experiment was conducted before and after applying the pomegranate seed extract scalp tonic, and the measurements were made in an environment where the laboratory temperature was 24–25°C and the relative humidity was 40–60%.

3) Monangdang Hair count

The number of hairs per hair follicle was measured using the same method as the number of hairs per unit area (3.6 mm × 2.8 mm) of the scalp. To ensure the reliability of the measurement value, the measurements were repeated three times for each subject and the average was calculated. In addition, in the case of the average value with a deviation range exceeding 30% in each group, the mean ± standard deviation (Mean ± SD) was calculated using SPSS for Windows version 23.0. The measurements were conducted twice, before and after applying the pomegranate seed extract scalp tonic, and the measurements were taken in an environment with a laboratory temperature of 24–25°C and a relative humidity of 40–60%.

3. Differences in hair test amount according to pomegranate seed extract

1) Number of hairs

Hair on a healthy scalp has a hair cycle of about 80-90% growth phase, 1-2% regression phase, and 10-20% resting phase, with about 175-300 terminal hairs per 1㎠ and 50-100 physiological hairs falling out per day.

Hair growth and hair loss occur through the interaction between hair papilla cells and hair matrix cells, and the process goes through a growth phase of 4-6 years, a regression phase of 2-3 weeks, and a resting phase of 2-3 months, during which the hair follicle and hair papilla separate, growth completely stops, existing hair falls out, and new hair is generated.

In order to compare the effect of pomegranate seed extract through changes in the number of hairs before and after the experiment in hair care using an anti-hair loss mist containing pomegranate seed extract, the experimental group was divided into an experimental group and a control group, and the experimental group was treated with a pomegranate seed extract scalp tonic.

The number of hairs in the control group slightly increased from 16.20±2.15 before the experiment to 16.40±2.55 after the experiment, and there was no significant difference between the groups (p>.05). The experimental group showed an increase from 15.10±1.60 before the experiment to 19.90±1.84, and there was a significant difference between the groups (p<.001). In addition, the number of hairs in the control group (16.20±2.15) and the experimental group (15.10±1.60) before the experiment showed a difference but was not significant between the two groups (p>.05), and the number of hairs in the control group (16.40±2.55) and the experimental group (19.90±1.84) after the experiment treated with pomegranate seed extract showed a significant difference (p<.01). In other words, it was found that the experimental group that applied the pomegranate seed extract scalp tonic showed an increase in the number of hairs of approximately 32%.

Therefore, it was confirmed that hair loss prevention mist containing pomegranate seed extract is a good way to increase the number of hairs in hair care to prevent hair loss.

2) Hair thickness

The thickness of hair varies greatly depending on the type, from 0.04 to 0.12 mm (John, 1997). Soft hair is about 0.06 mm and normal hair is 0.075 to 0.08 mm.

In a quantitative analysis of Korean scalp hair characteristics using a phototrichogram, hair diameter decreased over time in a study measuring normal subjects for two months.

The physicochemical characteristics and morphological changes of hair in the elderly male and female population with aging were reported to show that the thickness and tensile strength of hair decrease with aging. Hair was found to be thicker in women in their 20s than in other women, and hair in women in their 40s or older was found to be thinner than in other women.

This is because women in their early 20s have thick terminal hair due to the influence of hormones, and when they become adults, their hair does not remain terminal hair, but rather gradually thins and returns to vellus hair around the age of 30, so it can be inferred that hair becomes thinner than other age groups when they are in their 40s or older.

In order to compare the effect of pomegranate seed extract through changes in hair thickness before and after the experiment in hair care using anti-hair loss mist containing pomegranate seed extract, the experimental group was divided into an experimental group and a control group, and the anti-hair loss mist containing pomegranate seed extract was applied to the experimental group. After 12 weeks of clinical trials, the following results were obtained.

The hair thickness of the control group was similar from 0.05±0.00 before the experiment to 0.05±0.00 after the experiment, and there was no significant difference between the groups (p>.05). The experimental group showed an increase from 0.05±0.01 before the experiment to 0.06±0.05, and there was a significant difference between the groups (p<.01). In addition, the hair thickness of the control group (0.05±0.00) and the experimental group (0.05±0.01) before the experiment was not significant between the two groups (p>.05), and the hair thickness of the control group (0.05±0.00) and the experimental group (0.060±0.05) after the experiment treated with the pomegranate seed extract showed a significant difference (p<.001). In other words, it was found that the experimental group that applied the pomegranate seed extract scalp tonic showed an increase in hair thickness of about 20% more.

Therefore, it was confirmed that hair loss prevention mist containing pomegranate seed extract is a good way to increase hair thickness in hair care to prevent hair loss.

3) Monangdang Hair count

The human body generally has about 5 million hair follicles formed during the fetal period, of which about 100,000 are present on the scalp. After birth, no new hair follicles are formed, but only their size changes under the influence of androgens.

In the case of the scalp that protects the head, the hair roots and sweat glands (minor sweat glands) are developed, and it is a part that protects the brain from external pressure or impact, and is deeply involved in hair production that expels heavy metals from the body. In other words, the hair present in the scalp pores is a part that is sensitive to the health of the scalp, and there are countless pores, about 100,000, in the scalp tissue, and each pore has from 1 to 4 hairs, which are called 'hair follicle units'.

In order to compare the effect of pomegranate seed extract through the change in the number of hairs per hair follicle before and after the experiment in hair care using hair loss prevention mist containing pomegranate seed extract, the experimental group was divided into an experimental group and a control group, and the experimental group was treated with pomegranate seed extract scalp tonic. After 12 weeks of clinical trials, the following results were shown.

The number of hairs per follicle in the control group slightly decreased from 1.54±0.33 before the experiment to 1.44±0.29 after the experiment, and there was no significant difference between the groups (p>.05). The experimental group showed an increase from 1.54±0.34 before the experiment to 1.89±0.30, and there was a significant difference between the groups (p<.05). In addition, there was no significant difference between the number of hairs per follicle in the control group (1.54±0.33) before the experiment and the number of hairs per follicle in the experimental group (1.54±0.34) (p>.05), and there was a significant difference between the number of hairs per follicle in the control group (1.44±0.29) and the number of hairs per follicle in the experimental group (1.89±0.30) after the experiment treated with pomegranate seed extract (p<.01). That is, the experimental group that applied the pomegranate seed extract scalp tonic showed an increase of 23% from 1.54±0.34 to 1.89±0.30, indicating an increase in the number of hairs per hair follicle.

Therefore, it was confirmed that hair loss prevention mist containing pomegranate seed extract is a good method for increasing the number of hairs per hair follicle in hair care to prevent hair loss.

It was confirmed that applying a hair loss prevention mist containing pomegranate seed extract can improve various scalp and hair problems by making the scalp and hair condition healthy for middle-aged women in their 40s to 60s. In addition, it seems that using pomegranate seed extract to manage the scalp and hair of menopausal women will help improve negative perceptions of appearance and help them enjoy a satisfactory old age.

Claims (2)

A method for producing pomegranate seed extract for preventing hair loss, comprising a drying step for drying pomegranate seeds, a crushing step, a dissolution extraction step, and a concentration step,
The above drying step dries the pomegranate seeds at a temperature of 40℃ for 15 hours.
The above grinding step grinds dried pomegranate seeds at a temperature of 100℃ for 30 minutes.
The above dissolution extraction step is performed by maintaining the temperature of the aqueous solution mixed with pomegranate seed powder and 70% ethanol in a ratio of 1:3 at 80°C and repeating extraction and filtration three times for 3 hours while cooling the 80°C aqueous solution under reflux to extract the filtrate.
A method for producing a pomegranate seed extract for preventing hair loss, characterized in that the above concentration step comprises concentrating the extracted filtrate under reduced pressure at 50°C to obtain a pomegranate concentrate. 89 to 91 wt% of distilled water containing no organic or inorganic matter,
2-4% by weight of ethanol, a disinfectant that removes bacteria and fungi,
2 to 4 wt% of triethanolamine, which neutralizes fatty acids, adjusts and buffers pH, and dissolves oil;
Polysorbate 20 0.5 to 2 wt%, which evenly disperses liquids or solids that do not mix well with each other in a liquid,
Polysorbate 80, which acts as an emulsifier and stabilizer and uniformly disperses liquids or solids that do not mix well into a liquid, 0.5 to 2 wt%,
0.5~2% increase in glycerol, which acts as a moisturizer and prevents deterioration of other substances,
0.5 to 1 wt% of poloximer, which increases the solubility of hydrophobic and oily substances or increases the miscibility of two substances with different hydrophobic properties,
A mist composition for preventing hair loss containing pomegranate seed extract, comprising 1 to 2 wt% of pomegranate seed extract having estrogen and anti-estrogen effects.